Background: Pulmonary Embolism (PE) is a potentially life-threatening condition that is characterized by one or several blood clots blocking the arteries in the lungs. The existing diagnostic tools for PE have their shortcomings, highlighting the importance of investigating new diagnostic methods. The development of non-invasive methods to collect microparticles from exhaled breath has opened possibilities to explore new potential biomarkers. SensAbues® is a sampling device that utilizes electrostatic filters to capture microparticles from the exhaled breath. The objective of this project was twofold: firstly, to assess the suitability of SensAbues® sampling device for a future proteomics study where the goal is to identify biomarkers for PE; and secondly, to evaluate the efficacy of various extraction solutions in retrieving proteins from the electrostatic filters. Materials and methods: Samples were collected from three healthy volunteers using the SensAbues® device. The electrostatic filters were then extracted using either PBS or 15% ethanol and the protein content was then estimated using a modified Bradford method. Additionally, two blank SensAbues® filter extracts, from PBS and 15% ethanol were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results: The attempts to evaluate extraction solutions using the Bradford method were unsuccessful, as all the samples yielded negative values. The filter-blank extracts analyzed with LC-MS/MS contained a significant amount of polyethylene glycols of varying sizes. Conclusion: The polyethylene glycols from the SensAbues® filters may have interfered with the Bradford method. Polyethylene glycols can also interfere with proteins, making the SensAbues® sampling device unsuitable for the prospective proteomics study.

The work presented in this thesis is concerned with development of spectroscopic detection methods based on absorption spectroscopy using semiconductor lasers, with particular ref- erence to the field of medical diagnostics through breath analysis. The first part of this thesis deals with the design and testing of a prototype analyser for simultaneous monitoring of the exchange gases O₂ , CO₂ and H₂O in breath. The aim of this analyser is to provide information required to monitor respiration, with potential use in intensive care monitoring or during anaesthesia. The relatively high concentrations of these gases in breath and read- ily available diode laser sources make detection in the near-infrared (NIR) ideal. However, the relatively weakly absorbing A-band O₂ transitions at 760 nm require the application of a sensitive spectroscopic method, cavity enhanced absorption spectroscopy (CEAS). In contrast, CO₂ and H₂O are monitored using direct single pass absorption spectroscopy, with transitions arising from the 2ν₁ + ν₃ band at 2 μm and ν₁ + ν₃ band at 1.3 μm, respectively. It has been demonstrated that these gases can be detected simultaneously over a short pathlength (2.74 - 4 cm) in the respiratory flow by combining various spectroscopic methodologies and real-time data analysis. This analyser is shown to offer a viable alter- native for monitoring respiration, exhibiting absolute detection limits of changes of 0.26 % O₂ , 0.02 % CO₂ and 0.003 % H₂O with a 10 ms time resolution, which are comparable to current mass spectrometry based methods, but without their inherent delays. Following this, investigations into the detection of the main gas constituents in breath in the NIR employing noise-reduction modulation based spectroscopic techniques, namely wavelength and frequency modulation (WMS and FMS respectively) are also reported. The described WMS studies on water at 1.37 μm provide a demonstration of conventional WMS detection, as well as a “proof-of-principle” example of a relatively new approach to calibrating the non-absolute information obtained from a WMS absorption signal. Typically WMS spectra are calibrated using mixtures of known gas concentrations or an absolute direct absorption spectrum where possible. In this work however, a self-calibrating method, the phasor decomposition method (PDM), is employed and the returned concentration from this calibration is compared to direct absorption measurement. From this, the calculated concentration using the PDM is found to differ by 9 % from the concentration value obtained by direct absorption, providing an alternative method of calibration for when direct absorption measurements are not possible. The use of FMS in the NIR is also demonstrated as a potential alternative to CEAS for monitoring O₂ at 760 nm. FMS detection is performed on atmospherically broadened O₂ and a time-normalised α_min(t) of 2.45 ×10⁻⁶ cm⁻¹ s^1/2 is obtained, which is two orders of magnitude less sensitive than the value of α_min(t) = 2.35 ×10⁻⁸ cm⁻¹ s^1/2 obtained with CEAS. This combined with the experimental requirements of an FMS system, make its use for detection of O₂ a less practicable option compared to CEAS for real-time breath analysis. The latter work in this thesis involves a change in focus to detection of trace gases in breath in the mid-infrared (MIR). The move of spectroscopic detection to the MIR exploits the larger absorption cross-sections available in this region, and to achieve this, a relatively new form of semiconductor laser, the quantum cascade laser (QCL) is used. The design of a continuous wave QCL spectrometer at 8 μm and its operating characteristics are demon- strated and improvements in its performances are also discussed. This QCL system is then utilised to demonstrate the potential of monitoring species in breath, namely the narrow- band absorber methane and the broadband absorber acetone, taking into consideration the potential interference from other absorbing species in breath and the different spectroscopic characteristics exhibited by these molecules. Finally, the potential to further improve the sensitive detection of trace gases in breath in the MIR is also investigated with studies on the use of CEAS and multipass cells. In this work, the molecule of interest is the biomarker OCS, using transitions of the 2ν₂ band at 1031 cm⁻¹ , that are probed using a 10 μm QCL. The application of CEAS in the MIR is not as well developed as in the NIR, and the experimental consequences of using optical cavities at these wavelengths, where equipment tends to be more limited, are investigated and sensitivities discussed in the context of other literature. The experimental procedure of optimising a cavity for CEAS using the off-axis alignment method is also studied in detail, as well as the addition of WMS to further improve the signal quality. An effective absorption pathlength of ∼ 100 m was achieved in the cavity, with a bandwidth reduced α_min(BW) of 1.7 ×10⁻⁷ cm⁻¹ Hz^−1/2 using WMS CEAS achieved. With the poorer quality optics and limitations in equipment in the MIR for CEAS experiments, the use of a multipass cell, a 238 m Herriott cell, is also investigated as an alternative to the use of an optical cavity at 10 μm. Detection of OCS using direct absorption and WMS is demonstrated in the Herriott cell, achieving α_min(BW) = 2.03×10⁻⁸ cm⁻¹ Hz^−1/2 using WMS. This shows an improvement in sensitivity compared to WMS CEAS, and also shows the potential for future work on biomarker detection, as it approaches the ∼ ppb levels required for breath analysis.

This research was aimed at the development and validation of new in vitro methods capable of predicting in vivo drug deposition from dry powder inhalers, DPIs, in lung-normal human adults. Three physical models of the mouth, throat and upper airways, MT-TB, were designed and validated using the anatomical literature. Small, medium and large versions were constructed to cover approximately 95% of the variation seen in normal adult humans of both genders. The models were housed in an artificial thorax and used for in vitro testing of drug deposition from Budelin Novolizer DPIs using a breath simulator to mimic inhalation profiles reported in clinical trials of deposition from the same inhaler. Testing in the model triplet produced results for in vitro total lung deposition (TLD) consistent with the complete range of drug deposition results reported in vivo. The effect of variables such as in vitro flow rate were also predictive of in vivo deposition. To further assess the method’s robustness, in vitro drug deposition from 5 marketed DPIs was assessed in the “medium” MT-TB model. With the exception of Relenza Diskhaler, mean values for %TLD+SD differed by only < 2% from their literature in vivo. The relationship between inhaler orientation and in vitro regional airway deposition was determined. Aerosol drug deposition was found to depend on the angle at which an inhaler is inserted into the mouth although the results for MT deposition were dependent on both the product and the formulation being delivered. In the clinic, inhalation profiles were collected from 20 healthy inhaler naïve volunteers (10M, 10F) before and after they received formal inhalation training in the use of a DPI. Statistically significant improvements in Peak Inhalation Flow Rate (PIFR) and Inhalation Volume (V) were observed following formalized training. The shapes of the average inhalation profiles recorded in the clinic were found to be comparable to the simulated profiles used in the in vitro deposition studies described above. In conclusion, novel in vitro test methods are described that accurately predict both the average and range of aerosol airway drug deposition seen from DPIs in the clinic.

O uso nocivo do álcool acarreta problemas de saúde para o usuário e prejuízos na sociedade. Vários espécimes biológicos podem ser utilizados para verificar a exposição a bebidas alcoólicas. No trabalho foi investigada a utilização da saliva na determinação de etanol através das técnicas: enzimática Q.E.D. A-150 de triagem - e cromatografia em fase gasosa, após separação por aquecimento, Headspace de confirmação. O método foi padronizado, validado e aplicado em amostras de saliva provenientes de voluntários que ingeriram de maneira controlada bebida alcoólica. Em todas as amostras analisadas foi possível detectar a concentração de etanol, observando a existência de uma diferença entre os respectivos resultados das duas técnicas. Os resultados interindividuais apresentaram grande variabilidade nos mesmos tempos de coleta, apesar da ingesta equivalente de etanol. No estudo foi verificado que a saliva é uma matriz alternativa adequada para verificar a concentração de etanol em indivíduos expostos às bebidas alcoólicas. / The abuse of alcohol leads to health injury and causes damage to the society. Several biological matrices can be used to monitor exposure to alcoholic beverages. In this work, saliva was used in the determination of ethanol for both techniques: enzymatic Q.E.D. A-150 screening test -, and headspace-gas chromatography - confirmation test -. The method was developed, validated and used for the analysis of saliva samples from volunteers who ingested alcoholic beverages in a controlled schedule. Ethanol was detected and in all samples, noticing a difference between the results of both techniques. Inter-volunteers results showed high variability for the same collection times, although the amount of ethanol ingested was equivalent. These results suggest that saliva is a good option for the in field determination of ethanol in exposed individuals to alcoholic beverages.

A dissertação se situa no campo de pesquisa de métodos não invasivos de análise clínica, mais especificamente, de prognóstico da gravidade de insuficiência cardíaca, IC, com base em bioindicador presente no ar exalado por pacientes, em estudo no IncorHCFMUSP. O bioindicador mais relevante encontrado por GC-MS foi a acetona (propanona). Para a coleta do ar exalado desenvolveu-se dispositivo que compreende frasco borbulhador (impinger) com difusor, contendo 5 mL de água, imerso em banho de gelo e acoplado a saco plástico para definição de volume amostrado (7,6 L). A preservação de amostras deu-se por congelamento a -80ºC. Para a determinação do analito, escolheu-se método espectrofotométrico (474 nm) baseado na reação da acetona com salicilaldeído em meio básico, apresentando limite de detecção de 0,3 mg/L em fase líquida e 0,3 &#181;g/L em ar exalado. Foram realizados estudos de eficiência de coleta por geração de atmosfera controlada. Resultados iniciais obtidos com amostras coletadas de pacientes portadores de IC pelos colaboradores do Incor indicam a potencialidade do bioindicador e dos equipamentos e métodos desenvolvidos para acetona nesta dissertação. / The dissertation fits in the research field of noninvasive clinical analysis, more specifically, the prognosis of the severity of heart failure, HF, based on a biomarker found in the air exhaled by patients, in a study at InCor-HCFMUSP. The most relevant biomarker found by GC-MS was acetone (propanone). For the breath collection a device was developed which comprises a scrubber flask (impinger) with diffuser, containing 5 mL of water, immersed in an ice bath and attached to a plastic bag for delimitation of the sample volume (7,6 L). For the determination of acetone a spectrometric method (474 nm) was chosen based on a reaction of acetone with salicylaldehyde in alkaline medium, with detection limits of 0,3 mg/L in liquid phase and 0,3 &#181;g/L in breath. Preliminary studies made with samples collected from HF patients by the co-workers from InCor indicate the potential of the identified biomarker and the suitability of the equipment and methods developed in this dissertation for its analysis.

Introdução: As doenças cardíacas ou do aparelho circulatório são as principais causas de morte na população brasileira, sendo responsável por um terço dos óbitos. Embora o tratamento conservador seja amplamente adotado, grande parte das doenças cardiovasculares necessita e tem como tratamento a cirurgia cardíaca, da qual as mais comuns são a cirurgia de revascularização do miocárdio (CRVM) e de troca valvar. Atualmente existem diversos estudos avaliando a função pulmonar ou a capacidade funcional para o acompanhamento do tratamento conservador ou em situação pré-operatória, porém estudos avaliando e correlacionando essas variáveis no pré-cirúrgico são escassos na literatura científica não sendo possível averiguar a existência da relação entre os níveis de nitrito e nitrato e o desempenho nos testes de capacidade funcional. Objetivo: Avaliar a correlação nitrito/nitrato (NOx) do condensado do exalado pulmonar (CEP) e a distância caminhada nos testes de caminhada de 6 minutos (TC6) e shuttle walk test (SWT), além de verificar se a utilização de medicações contendo betabloqueadores, nitrato ou enzima conversora de angiotensina (IECA) podem influenciar nas concentrações de nitrito/nitrato no CEP e nas distâncias caminhadas no TC6 e no SWT e comparar seu comportamento entre valvopatas e coronariopatas no pré operatório de cirurgia cardíaca. Metodologia: Foram selecionados 73 pacientes, dos quais 28 eram coronariopatas e 45 valvopatas, de ambos os sexos, com idade entre 20 e 80 anos, feita coleta do CEP para análise do NOx e submetidos ao TC6 e ao SWT para avaliação da distância caminhada. Resultados: Na análise das concentrações de NOx do CEP não foi encontrada diferença estatisticamente significativa entre as medicações utilizadas via oral contendo nitrato (dinitrato de isossorbida), IECA (captopril ou enalapril) e/ou betabloqueador ou entre os pacientes do grupo coronariopata e grupo valvopata (p>0,05). Houve diferença estatisticamente significativa no TC6 nos valores de frequência cardíaca (FC) inicial, final e repouso e no valor referente à dispnéia final, e no SWT nos valores de FC inicial, final e repouso, na pressão arterial sistólica (PAS) inicial e no valor referente à dispnéia final entre os grupos coronariopata e valvopata. No entanto, foram interrompidos 4 TC6 e 7 SWT do grupo coronariopata devido ao desencadeamento de dor precordial, fato não ocorrido no grupo valvopata, apresentando nível de significância. A FC inicial e de repouso no TC6 e a FC final no SWT dos pacientes que não ingeriram nenhum medicamento antes do teste é diferente em relação à dos pacientes que fizeram uso de quaisquer medicamentos descritos antes do teste sugerindo a ação de medicamentos que modulam a FC como os betabloqueadores. Foi constatado correlação positiva moderada entre o NOx do CEP e os valores de PAS inicial e final de coronariopatas e correlação negativa fraca a moderada entre o NOx do CEP e os valores de FC final e de dispnéia final nos valvopatas no TC6, além da correlação positiva moderada entre o NOx do CEP e os valores de PAS inicial de indivíduos coronariopatas e correlação negativa fraca a moderada entre o NOx do CEP e os valores de FC inicial nos valvopatas no SWT. Foi identificado correlação negativa moderada na distância caminhada no TC6 e no SWT apenas nos indivíduos coronariopatas. Conclusões: Não foram identificadas influências do uso de medicações betabloqueadoras, nitrato ou IECA sobre a dosagem de NOx ou a distância caminhada em nenhum dos testes e em nenhum dos grupos, no entanto coronariopatas com níveis mais elevados de NOx apresentaram uma distância deambulada menor. / Introduction: Heart disease or circulatory system diseases are the leading causes of death in our population, accounting for one third of deaths. Although conservative treatment is widely adopted, part of cardiovascular disease treatment demands heart surgery, of which the most common are coronary artery bypass grafting (CABG) and valve replacement. Currently there are many studies evaluating lung function or functional capacity for monitoring of conservative treatment or preoperative situation, but studies assessing and correlating these variables in pre surgical are rare in the literature is not possible to ascertain the existence of the relationship between levels of nitrite and nitrate (NOx) in exhaled breath condensate (EBC) and performance in functional ability tests. Methods: were selected 73 patients, 28 with coronary artery diseas and 45 with heart valve disease of both genders, aged 20 to 80 years old. The EBC was collected for analysis of NOx and performed the six minutes walking test (6MWT) and shuttle walk test (SWT) to evaluate the distance walked. Results: the analysis of the concentrations of NOx in EBC has no statistically significant difference found between the medications used, such as oral nitrate (isosorbide dinitrate), ace inhibitors (captopril or enalapril) and/or beta-blockers in both coronary artery disease and heart valve disease (p> 0.05). There was a statistically significant difference in 6MWT for HR initial, final and rest and final dyspnea, and in SWT for HR initial, final and rest, the initial SBP and the final dyspnea between the coronary artery disease e heart valve disease. However, were interrupted 4 6MWT and 7 SWT from coronary artery disease related to chest pain, this fact was not related in patients with valve disease, presenting a significance level. The initial and resting HR during the 6MWT and in SWT final HRr patients not ingesting any medication before the test is different from patients who used any medications before the test, suggesting the action of drugs that modulate the HR, such as beta blockers. Moderate positive correlation was found between the NOx in EBC and SBP initial and final in coronary disease and weak to moderate correlation between the NOx in EBC and final HR and final dyspnea in 6MWT from heart valve disease. Besides the positive moderate correlation between NOx in the EBC and the SBP initial in coronary disease during 6MWT and weak to moderate negative correlation between NOx in the EBC and the HR initial in valve disease during SWT. Moderate negative correlation was identified in the distance walked during the 6MWT and SWT only between individuals with coronary artery disease. Conclusions: the use of beta-blocker drugs, ace inhibitors or nitrate evidenciates no influence on the NOx or on the distance walked in any tests, and none of the groups, however, individuals with coronary artery disease showed higher NOx levels and performed lower distance walked.

Introdução: A má digestão de lactose detectada por testes respiratórios é bastante comum em pacientes com síndrome do intestino irritável (SII), e alguns pacientes com SII referem exacerbar seus sintomas após a ingestão de leite e produtos lácteos. A fim de uma melhor percepção sobre este tema, este trabalho investigou os efeitos da dieta isenta de lactose em pacientes com SII e teste respiratório de hidrogênio para avaliação da digestão de lactose normal ou anormal. Objetivo: Avaliar os efeitos de uma dieta isenta de lactose em pacientes de SII com ou sem teste positivo para má digestão de lactose (teste respiratório). Métodos: 80 pacientes atendidos no Ambulatório de Doenças Funcionais Intestinais do Serviço de Gastroenterologia Clínica do HCFMUSP foram recrutados para o estudo prospectivo. Os pacientes preencheram os critérios de Roma III para SII. Durante o ensaio do teste respiratório de hidrogênio todos os sintomas desencadeados pela ingestão de lactose foram anotados. Ambos os grupos de pacientes com e sem má digestão de lactose, foram submetidos a uma dieta isenta de lactose durante 4 semanas. A história prévia de intolerância ao leite e derivados também foi documentada. No final de cada semana, os pacientes foram solicitados a informar a respeito dos seus sintomas, e os resultados, foram devidamente marcados de acordo com a sua intensidade (Escore Global). Além disso, foi também obtida a Avaliação Global do Paciente no final do tratamento (AFG). Resultados: 33/80 (41,25%) pacientes com SII tiveram um teste respiratório anormal para a má digestão de lactose, e 47/80 dos pacientes (58,75%) com SII tiveram um teste respiratório normal (ns). Não houve diferença significativa na freqüência de queixas de intolerância prévia ao leite e derivados em ambos os grupos. No entanto, mais pacientes no grupo com má digestão de lactose referiram sintomas durante o teste respiratório com 25 g de lactose do que o grupo com digestão normal a lactose (p <0,001). Ambos os grupos igualmente melhoraram após uma dieta isenta de lactose, quando avaliados pelo escore geral e pelo AFG. Conclusões: Ambos os grupos de pacientes com SII, com e sem má digestão de lactose, melhoraram após 4 semanas de dieta isenta de lactose. Os nossos resultados sugerem que outros mecanismos, além da intolerância à lactose, devem ser levados em conta para explicar a melhora com uma dieta isenta de lactose em pacientes com SII / Introduction: Lactose intolerance detected by respiratory tests is rather common in patients with irritable bowel syndrome(IBS); also, some IBS patients refer their symptoms exacerbate after ingestion of milk and dairy products. In order to gain some insight into this topic we investigated the effects of a lactose-free diet in IBS patients with and without lactose intolerance diagnosed by means of the hydrogen respiratory test. Objective: To evaluate the effects of a lactose-free diet in IBS patients with and without lactose intolerance. Methodos: 80 patients who attended the outpatient clinic at the University of São Paulo School of Medicine Hospital, Brazil, were recruited for this prospective study. Patient met the Rome III criteria for IBS. During the hydrogen respiratory test all the symptoms triggered by lactose ingestion were taken down. Both groups of patients, with and without lactose intolerance, were submitted to a lactose-free diet for 4 weeks. Previous history of milk/dairy products intolerance was documented. At the end of each week, patients were asked to inform about their symptoms and the results were duly scored according to their intensity (Global Score). Also, the Patient Final Global Evaluation of the Treatment (PFGET) was obtained as well. Results: 33/80 (41.25%) IBS patients had an abnormal respiratory test for lactose intolerance; 47/80 (58.75%) IBS patients had a normal respiratory test (n.s.). There was no significant difference in the frequency of previous complaints after milk/dairy products ingestion in both groups. However, more patients in the lactose intolerant group referred symptoms during the respiratory test with 25 g of lactose than the lactose tolerant group (p<0,001). Both groups similarly improved after a lactose-free diet when evaluated by both the Global Score and the PFGET. Conclusions: Both groups of IBS patients, with and without lactose intolerance, improved after a 4-week lactose-free diet. Our results suggest that other mechanisms, in addition to lactose intolerance, should be taken into account to explain why IBS patients improve with a lactose-free diet

Made available in DSpace on 2016-03-15T19:47:36Z (GMT). No. of bitstreams: 1 Maria Eloisa de Souza Ivan.pdf: 1241876 bytes, checksum: c243992308e7c5e5c81555fd3183f288 (MD5) Previous issue date: 2015-02-27 / Started in 1974, the posthumous book Um sopro de vida (pulsações), by Clarice Lispector, came to public in 1978. Written "in agony" (Borelli, 1981), the work resumes introspective density featuring Clarice's writing, repeats the existential paradox life and death so recurrent in her work, reveals itself as a work of beginning and end, one and multiple, finally, leading us by several known ways, or not. Recognizing that reading involves an empathetic relationship and motivated to continue the master's research, also about Clarice's work, I felt compelled to face the reading of such a dense work and to seek still little trodden paths. Thus, in this project, we have resumed aspects which have not been explored in our master's research for not being part of the objectives of that work, but which were perceived as relevant for further study. Among these aspects, we mention the manifestation of the double as a structural element of Um sopro de vida (pulsações) narrative, 1978, chosen as the corpus of this research. The theme of the double has always been present in the Western culture, varying according to the historical moment. Renovated in modernity, the myth of the double also reveals the man as a shattered person, having as one of its possible representations the division of the "I" in an "ego" and an "alter ego". To better understand the relationship of the double with the work, we can say that many centuries separate the Greek legend, told by Aristophanes, and from the book Um sopro de vida (pulsações), by Clarice Lispector. The legend seeks to explain the need of human beings to ever get the half that fulfills them; Clarice's work embodies, among others, the man's questioning in search of himself, reflecting on the "who am I?", "who is the other?". In this way, describing and analyzing the figure of the narrator-Author, establishing a counterpoint between narrator-Author/Ângela Pralini and observing the process of building the character like an image that is constituted "by" the other makes up the specific objectives of this study; it is also our purpose to investigate and analyze the effects of discursive meanings that recover the images of God, of the "you", of the mirror and the death in relation to the narrator-Author and Ângela Pralini. Guiding us, then, through reflections, by Rank, Clément Rosset, Bravo, Bakhtin, among others, the research presented here proposes, as a questioning, to investigate how the image of the double emerges as a structuring element of Clarice's narrative, more specifically in Um sopro de vida (pulsações), and to what extent this discursive feature manifests itself in the narrator-Author/Ângela Pralini counterpoint, his double, also investigating how this narrator is manifested in personal oscillations, in the expansion of the narrating function, in the lexical choices, in the adopted bias: who is the double of whom? / Iniciado em 1974, o livro póstumo Um sopro de vida (pulsações), de Clarice Lispector, vem a público em 1978. Escrito &#8213;em agonia&#8214; (Borelli, 1981), a obra retoma a densidade introspectiva que caracteriza a escrita clariceana, repete o paradoxo existencial vida e morte tão recorrente em sua obra, revela-se como obra de princípio e fim, uno e múltiplo, enfim, nos guia por vários caminhos conhecidos, ou pouco explorados. Reconhecendo que leitura envolve relação empática e motivada a continuar a pesquisa de mestrado, também acerca da obra clariceana, senti-me provocada a enfrentar a leitura de obra tão densa e buscar esses caminhos ainda pouco trilhados. Assim, neste projeto, retomamos aspectos os quais não foram explorados na nossa pesquisa de mestrado por não fazerem parte dos objetivos daquele trabalho, mas que foram percebidos como relevantes para um estudo posterior. Dentre esses aspectos, citamos a manifestação do duplo como elemento estruturante da narrativa Um sopro de vida (pulsações),1978, escolhida como corpus desta pesquisa. A temática do duplo sempre esteve presente na cultura ocidental, variando de acordo com o momento histórico. Renovado na modernidade, o mito do duplo também revela o homem estilhaçado, tendo como uma de suas possíveis representações a cisão do Eu em um &#8213;ego&#8214; e um &#8213;alter ego&#8214;. Para melhor entendermos a relação do duplo com a obra, podemos dizer que muitos séculos separam a lenda grega, contada por Aristófanes, do livro Um sopro de vida (pulsações), de Clarice Lispector. A lenda procura explicar a necessidade do ser humano de buscar eternamente a metade que o completa; a obra clariceana materializa, entre outros, o questionamento do homem em busca de si mesmo, refletindo acerca do &#8213;quem sou eu?&#8214; &#8213;quem é o outro?&#8214; Desse modo, compõem os objetivos específicos deste estudo descrever e analisar a figura do narrador-Autor, estabelecendo-se um contraponto entre narrador-Autor/Ângela Pralini, observando-se o processo de construção da personagem como imagem que se constitui &#8213;pelo&#8214; outro; também é nosso propósito investigar e analisar os efeitos de sentido discursivos que recobrem as imagens de Deus, do vós, do espelho e da morte em relação ao narrador-Autor e Ângela Pralini. Orientando-nos, então, pelas reflexões de Rank, Clément Rosset, Bravo, Bakhtin, entre outros, a pesquisa que aqui se apresenta propõe, como questionamento, investigar de que modo a imagem do duplo se materializa como um elemento estruturante da narrativa clariceana, mais especificamente em Um sopro de vida (pulsações), e em que medida esse recurso discursivo se manifesta no contraponto narrador-Autor/Ângela Pralini, seu duplo, investigando também como esse narrador se manifesta nas oscilações de pessoa, na ampliação da função de narrar, nas escolhas vocabulares, no viés adotado: quem é o duplo de quem?

O uso nocivo do álcool acarreta problemas de saúde para o usuário e prejuízos na sociedade. Vários espécimes biológicos podem ser utilizados para verificar a exposição a bebidas alcoólicas. No trabalho foi investigada a utilização da saliva na determinação de etanol através das técnicas: enzimática Q.E.D. A-150 de triagem - e cromatografia em fase gasosa, após separação por aquecimento, Headspace de confirmação. O método foi padronizado, validado e aplicado em amostras de saliva provenientes de voluntários que ingeriram de maneira controlada bebida alcoólica. Em todas as amostras analisadas foi possível detectar a concentração de etanol, observando a existência de uma diferença entre os respectivos resultados das duas técnicas. Os resultados interindividuais apresentaram grande variabilidade nos mesmos tempos de coleta, apesar da ingesta equivalente de etanol. No estudo foi verificado que a saliva é uma matriz alternativa adequada para verificar a concentração de etanol em indivíduos expostos às bebidas alcoólicas. / The abuse of alcohol leads to health injury and causes damage to the society. Several biological matrices can be used to monitor exposure to alcoholic beverages. In this work, saliva was used in the determination of ethanol for both techniques: enzymatic Q.E.D. A-150 screening test -, and headspace-gas chromatography - confirmation test -. The method was developed, validated and used for the analysis of saliva samples from volunteers who ingested alcoholic beverages in a controlled schedule. Ethanol was detected and in all samples, noticing a difference between the results of both techniques. Inter-volunteers results showed high variability for the same collection times, although the amount of ethanol ingested was equivalent. These results suggest that saliva is a good option for the in field determination of ethanol in exposed individuals to alcoholic beverages.

<p>The aim of this study is to examine the knowledge and experience of the bad breath among high school students in Kristianstad. A further objective was to measure the amount of VSC (Volatile Sulfur Compounds) among those who perceive themselves to have a bad breath. A questionnaire with 21 closed questions was distributed to 120 high school students from Kristianstad municipality who were between the ages of 17-20. The results of this study revealed that most of the students had good knowledge of halitosis. The majority of all participants thought it was important to smell fresh in the mouth and also experienced most of their breath as very good or good. Only a few felt that they had bad breath. However, these persons did think their breath affect their everyday life. Furthermore, they have never felt embarrassed or caught in embarrassing situations due to their breath. The results from the clinical examination was carried out on 8 of the 120 students and was designed to measure the amount of sulphurous gases in the oral cavity which revealed that the average values that emerged in the examination below is the limited values for having a poor breath.</p> / <p>Syftet med denna studie var att undersöka kunskapen och upplevelsen av dålig andedräkt hos gymnasieelever i Kristianstad, ytterligare ett syfte var att mäta mängden VSC (reaktiva svavelföreningar)  hos dem som upplever sig ha dålig andedräkt. En enkät med 21 slutna frågor delades ut till 120 gymnasieelever som var mellan 17-20 år och studerade i Kristanstads kommun. Av resultatet i denna studie framkom att de flesta av gymnasieeleverna hade goda kunskaper om halitosis. Nästan alla 120 som besvarade enkäten tyckte att det var viktigt att lukta fräsch i munnen och dessutom upplevde de flesta sin andedräkt som mycket bra eller bra. Endast ett fåtal (14% ) kände att de ibland hade dålig andedräkt. Hos dessa  påverkades inte andedräkten deras vardagliga liv och de hade aldrig blivit generade eller hamnat i pinsamma situationer på grund av sin andedräkt. Resultatet från den kliniska undersökningen som gjordes på 8 av de 120 elever som besvarade enkäten där mängden av de svavelhaltiga gaserna i munhålan mättes visade att i genomsnitt ligger värden som framkom i undersökningen under gränsvärden för att man skall ha dålig andedräkt.</p>