Há diversas evidências epidemiológicas de correlações positivas entre indicadores de morbidade e mortalidade pulmonar e cardiovascular e aumentos na concentração atmosférica de MP2,5 (material particulado fino). O objetivo deste trabalho foi avaliar os efeitos da exposição subcrônica de MP2,5 sobre o tônus cardíaco autonômico, a inflamação pulmonar e sistêmica; o estresse oxidativo e a homeostase sanguínea, após oito semanas de repetidas instilações nasais de suspensão aquosa de MP2,5 da cidade de São Paulo em concentração ambiental. Dividiram-se os animais em dois grupos: salina e MP2,5 e avaliaram-se os seguintes parâmetros: frequência cardíaca (FC), variabilidade da frequência cardíaca (VFC), pressão arterial sistólica (PA), hemograma, contagem de plaquetas e reticulócitos, fibrinogênio plasmático, tempo de protrombina (TP), tempo de tromboplastina parcialmente ativada (TTPA), mielograma, citologia do lavado broncoalveolar (LBA), análise histopatológica e imuno-histoquímica (15-F2tisoprostano e -actina) de pequenas arteríolas pulmonares e coronarianas. Não houve alterações na FC e na PA (p > 0,05). Houve interação estatisticamente significante entre grupos e semanas em relação à VFC. O SDNN (desvio padrão dos intervalos R-R normais), a r-MSSD (raiz quadrada da média dos quadrados das diferenças sucessivas entre intervalos R-R normais adjacentes) e a AF (alta frequência) do grupo MP2,5 aumentaram significativamente na 7ª semana em comparação à 1ª semana (p < 0,05), enquanto a BF (baixa frequência) não se alterou (p > 0,05). A porcentagem de macrófagos no LBA do MP2,5 diminuiu significativamente (p < 0,05). Não se observaram alterações no sangue, mielograma e análise histopatólogica e imuno-histoquímica dos vasos (p > 0,05). Concluiu-se que a exposição subcrônica pela instilação nasal de suspensão aquosa de MP2,5 em concentração ambiental causou inflamação pulmonar tênue e alterou o equilíbrio cardíaco autonômico / There are several epidemiological evidences of positive correlation between indicators of pulmonary and cardiovascular morbidity and mortality and increases of PM2.5 (fine particulate matter) air concentration. The aim of this experiment was to evaluate the effects of subchronic exposure of PM2.5 on cardiac autonomic tone, pulmonary and systemic inflammation, oxidative strees and blood homeotasis of healthy rats after eight weeks of repeated nasal instillations of suspended PM2.5 from Sao Paulo city in environmental concentration. Rats were divided in two groups: saline and PM2.5. The following parameters were evaluated: heart rate (HR), heart rate variability (HRV), systolic blood pressure (BP), hemogram, platelets and reticulocytes count, plasmatic fibrinogen, prothrombin time (PT), activated partial thromboplastin time (APTT), bone marrow cells, bronchoalveolar lavage cells (BAL), histopathological and immunohistochemical analysis (15-F2tisoprostane and -actin) of pulmonary and coronary small arterioles. No changes were detected in HR and BP (p > 0.05). There were a statistically significant interaction between groups and weeks in relation to HRV. SDNN (standard deviation of normal RR intervals), r-MSSD (square root of the mean of the squared differences between adjacent normal RR intervals) and HF (high frequency) of PM2.5 group significantly increased on 7th week compaired to 1st week (p < 0.05), while LF (low frequency) did not alter (p > 0.05). BAL macrophages porcentage of PM2.5 group significantly decreased (p < 0.05). No alterations were observed in blood, bone marrow cells, histopathological and immunohistochemical analysis of vessels (p > 0.05). We concluded that subchronic exposure by nasal instillation of aquous suspension of PM2.5 in environmental concentration caused tenuous pulmonary inflammation and altered cardiac autonomic balance

L'objectif de cette thèse a été de développer une technologie pour l'assainissement des sols / sédiments sélénifères et d’étudier la réduction microbienne des oxy-anions de sélénium dans différentes conditions de respiration et de configurations du bioréacteur.Le sol sélénifère prélevé, dans les terres agricoles cultivées de blé au Pendjab (Inde), a été caractérisé et son lavage a été optimisé en faisant varier les paramètres tels que le temps de réaction, la température, le pH et le rapport liquide / solide. Afin de maximiser l'élimination et la récupération du sélénium à partir de ce sol, l'effet des ions compétiteurs et les composés oxydants comme les agents d'extraction pour le lavage du sol, ont également été étudiés. Bien que les agents oxydants aient montré une efficacité maximale d'élimination du sélénium (39%), la présence d'agents oxydants dans le lixiviat et le sol agricole peut augmenter le coût de leur post-traitement. Les plantes aquatiques, Lemma minor et Egeria densa ont été utilisées pour étudier la phyto-remédiation du lixiviat du sol contenant des agents oxydants. Cependant, l'efficacité d'élimination du sélénium par les plantes aquatiques a été significativement affectée par les fortes concentrations de ces agents oxydants dans le lixiviat du sol.Le rinçage du sol sélénifère a révélé un motif de migration du sélénium à travers la colonne du sol. La migration de la fraction de sélénium soluble de la couche supérieure vers la couche inférieure et sa réduction et son accumulation subséquentes dans les couches inférieures de la colonne de sol, ont été observées pendant le rinçage du sol. L'efficacité d'élimination du sélénium par la méthode de rinçage du sol a diminué avec une augmentation de la hauteur de la colonne. De plus, le lixiviat contenant des oxy-anions de sélénium obtenus à partir du lavage du sol, a été traité dans un réacteur UASB en faisant varier l'alimentation organique. Des effluents contenant moins de 5 μg de sélénium L-1 ont été obtenus, ce qui est conforme aux normes de l'USEPA pour la limite de rejet de sélénium dans les eaux usées.De plus, la bio-remédiation ex situ des oxy-anions de sélénium a été étudiée dans des conditions variables. Une bactérie aérobie (Delftia lacustris) capable de transformer le sélénate et le sélénite en sélénium élémentaire, mais aussi en composés d'ester de sélénium solubles jusque-là inconnus, a été isolée et caractérisée de manière fortuite. Alternativement, la bio-réduction anaérobie du sélénate couplé au méthane en tant que donneur d'électrons, a été étudiée dans des bouteilles de sérum et un filtre percolateur en utilisant des sédiments marins comme inoculum. Enfin, l'effet de la contamination d'autres oxy-anions chalcogènes, en plus du sélénium, a été étudié. La réduction simultanée de la sélénite et de la tellurite par un consortium microbien mixte ainsi que la rétention des nanostructures de Se et de Te biogènes dans l'EPS, ont été réalisées durant une opération de 120 jours dans un bioréacteur UASB / The aim of this Ph.D. was to develop a technology for the remediation of seleniferous soils/sediments and to explore microbial reduction of selenium oxyanions under different respiration conditions and bioreactor configurations.Seleniferous soil collected from the wheat-grown agricultural land in Punjab (India) was characterized and its soil washing was optimized by varying parameters such as reaction time, temperature, pH and liquid to solid ratio. In order to maximize selenium removal and recovery from this soil, effect of competing ions and oxidizing agents as chemical extractants for soil washing were also studied. Although oxidizing agents showed a maximum selenium removal efficiency (39%), the presence of oxidizing agents in the leachate and the agricultural soil may increase the cost of their post-treatment. Aquatic plants, Lemma minor and Egeria densa were used to study phytoremediation of the soil leachate containing oxidizing agents. However, the selenium removal efficiency by aquatic weeds was significantly affected by the high concentrations of these oxidizing agents in the soil leachate.Seleniferous soil flushing revealed the selenium migration pattern across the soil column. Migration of soluble selenium fraction from the upper to the lower layers and its subsequent reduction and accumulation in the lower layers of the soil column was observed during soil flushing. The selenium removal efficiency by the soil flushing method decreased with an increase in the column height. Furthermore, the soil leachate containing selenium oxyanions obtained from soil washing was treated in a UASB reactor by varying the organic feed. Effluent containing less than 5 μg L-1 selenium was achieved, which is in accordance with the USEPA guidelines for selenium wastewater discharge limit.Moreover, ex situ bioremediation of selenium oxyanions was studied under variable conditions. An aerobic bacterium (Delftia lacustris) capable of transforming selenate and selenite to elemental selenium, but also to hitherto unknown soluble selenium ester compounds was serendipitously isolated and characterized. Alternatively, anaerobic bioreduction of selenate coupled to methane as electron donor was investigated in serum bottles and a biotrickling filter using marine sediment as inoculum. Finally, the effect of contamination of other chalcogen oxyanions in addition to selenium was studied. Simultaneous reduction of selenite and tellurite by a mixed microbial consortium along with the retention of biogenic Se and Te nanostructures in the EPS was achieved during a 120-day UASB bioreactor operation

Les fines de lavage de carrières sont à l’heure actuelle peu valorisées. Pourtant, leurs caractéristiques physiques font de ces fines une matière première intéressante pour le domaine de la construction. L’objectif de cette thèse est de développer la formulation de produits préfabriqués non porteurs à base de fines de lavage, en considérant ces fines comme de la terre crue. Deux catégories de produits préfabriqués sont visées : un produit dit « lourd » (brique de terre comprimée) et un produit « léger » (carreau). Pour la valorisation en BTC, une étude de compacité par essai Proctor a permis de cibler la teneur en eau de fabrication et la masse volumique sèche à obtenir. Les performances mécaniques des briques ont été améliorées par surcompactage, renforcement granulaire et traitement au liant hydraulique. L’étude de formulation du carreau a associé les fines de lavage et la chènevotte. La consistance des mélanges à l’état frais a été étudiée avec le consistomètre VEBE. Les performances mécaniques en flexion et en compression des mélanges fibrés ont été mesurées. De la chaux et un superplastifiant ont été ajoutés dans la formulation pour satisfaire aux conditions de tenue mécanique. Les deux voies de valorisation ont été validées par mesures des performances physico-mécaniques sur produits fabriqués à l’échelle industrielle. La formulation optimale de BTC valorise près de 80% de fines de lavage et se classe dans la catégorie BTC 40 de la norme expérimentale XP13-901 (brique faiblement capillaire et résistante à la projection en eau). La formulation optimale de carreau se compose de 60% de fines de lavage, et répond aux exigences mécaniques des carreaux de plâtre. / Currently, few re-use ways are developed with clay fines from washing units in quarries. However, these clayey fines represent interesting materials for construction domain. This research work aims to develop non-load bearing precast products and to re-use these fines without thermal treatment as for unfired clay products. Two types of precast products are wished: a “heavy” product (compressed earth brick) and a “light” product (tile hemp-clay).For the CEB re-use way, the level of compaction has to be high. Proctor tests have been done to define the moisture content and dry density to obtain. Mechanical performances of bricks have been increased by overcompaction, granular reinforcement and lime treatment.For the tile re-use way, mixes with quarry fines and hemp have been studied. The behaviour of fresh material has been studied with VEBE consistometer. Flexural and compression strengths have been measured on hardened mixes. Lime and water-reducing agent have been necessary for a good mechanical behaviour.The two re-use ways have been validated by measuring mechanical and physical performances of products manufactured at industrial scale. The optimal mix for CEB includes almost 80% of quarry fines. The CEB is classed in BTC40 category according to experimental standard XP13-901 (brick with a low water absorption level and resistant to water spray). The optimal mix for tile includes 60% of quarry fines and satisfies mechanical requirements for gypsum blocks.

A constant flow of various pathogens enters the respiratory system on daily basis through the involuntary mechanism of breathing. Respiratory viral infections are common yet can be fatal in vulnerable populations. Respiratory syncytial virus (RSV) is one of the first and most common viruses that the human population acquire in the first two years of life. Despite the ability of most infants to recover from a RSV infection, many require hospitalization and, in few cases, die from such an infection. The pattern of seasonality of respiratory viruses also applies to RSV. In this work the temperature dependence of infectivity was studied in Hep-2 cells infected with RSV that had been incubated with bronchoalveolar lavage (BAL) fluid. The results indicate a temperature dependence of infectivity. Inhibition of the viral infectivity was observed at three different temperatures 37 ̊C, 40 ̊C and 42 ̊C. The inhibition appears to be linked to the appearance of large agglutinates that appear to reduce the infectivity of RSV. Such a study found that viral neutralization is dependent on a temperature-dependent agglutination reaction. The causality of agglutination formation requires further investigation in order to conclusively confirm the immunological component(s) of this reaction, and how temperature is contributing to this reaction.

This diploma thesis deals with the issue of left-handed retraining in the context of the research of Professor Miloš Sovák. It shows his contribution in this area and how he specifically contributed to the formation of teaching in the following years. The work is characterized by left-handedness, laterality. Examples of diagnostics of laterality of individual tests are given. In the theoretical part, the author also considers the breadth of the context of re-teaching not only the left-handed, where any violent re-learning can lead to consequences for life. In the practical part, a qualitative research was chosen, which is conducted on the basis of a semi-structured interview conducted with five respondents who answer open and closed questions about methods and approaches to their re-learning and their perception of the re- learning process. Research shows that there is no trait that is common to a smaller number of trainees, and therefore it is highly unlikely that there is a trait that would characterize a large group of trainees. However, given the research sample, it is not possible to draw a broader conclusion. KEYWORDS Milos Sovak, left-haned, retraining, reeducation, lavage, life story

Le processus d'hygiène des mains à l'urgence est une faille reconnue comme étant une source d'infections nosocomiales associées à la morbidité et la mortalité des patients. Malgré la mobilisation des organismes de santé pour résoudre le problème, le taux d'hygiène des mains (THM) chez les professionnels de la santé demeure insuffisant. Une analyse systémique du processus, en tenant compte de l'historique et du contexte socioculturel des établissements de santé, peut sans doute identifier les motifs susceptibles de freiner le sain processus d'hygiène des mains. Par conséquent, un laboratoire du changement (Lab-C) fut instauré dans une urgence d'un établissement du CHU de Québec en conformité avec les principes inhérents à la théorie historico-socioculturelle de l'activité (CHAT). Un sondage sur le processus d'hygiène des mains fut également réalisé auprès des professionnels de la santé afin de saisir leur compréhension du processus d'hygiène des mains et dans quelle mesure la pandémie à COVID-19 a influencé leurs habitudes. À la suite de ces analyses, nous constatons que les professionnels de la santé connaissent très bien le processus d'hygiène des mains. Ils disposent d'un niveau d'agentivité élevé leur permettant d'être réceptifs aux analyses et aux changements soutenant la modélisation d'un sain processus d'hygiène des mains. Cependant, malgré ces qualités et cette compréhension juste du processus, le THM demeure faible. Ceci peut s'expliquer notamment par l'impossibilité de prendre une pause pour effectuer le lavage des mains parce que les professionnels doivent maintenir une performance optimale dans une urgence qui fonctionne à plein régime. D'autre part, les principales raisons qui expliquent le faible THM sont la non-reconnaissance du besoin du lavage avant et après contact avec le patient ainsi que l'enjeu organisationnel dont notamment celui du matériel. Il appert que l'implantation d'un Lab-C dans une urgence est un bon outil technopédagogique et un bon agent modulateur du changement qui soutient l'analyse et la compréhension des problèmes systémiques trop souvent retrouvés dans nos établissements de santé. Il est proposé un outil technologique qui agit comme stimulus pour favoriser l'action du lavage des mains lorsque le professionnel pénètre ou sort de la bulle du patient. De plus, le patient peut stimuler l'action en contribuant au processus d'audit. / Hand hygiene process in the emergency department is recognized as a source of nosocomial infections associated with patient morbidity and mortality. Despite the mobilization of health organizations to solve the problem, the hand hygiene rate (THM) among health professionals remains insufficient. A systemic analysis of the process, considering the history and sociocultural context of healthcare facilities, would identify the recurring reasons that may hinder the healthy process of hand hygiene. Therefore, a change laboratory (Lab-C) was set up in an emergency room at the CHU de Québec in accordance with the principles inherent to Cultural Historical Activity Theory (CHAT). A hand hygiene process survey was also conducted among healthcare professionals to better understand the process and how the COVID-19 pandemic has influenced their hand hygiene habits. A hand hygiene process survey was also conducted among healthcare professionals to understand their opinion of the process and how the COVID-19 pandemic has influenced their hand hygiene habits. They have a high level of agency likely to challenge them in the modeling of a healthy hand hygiene process. However, despite these qualities and this correct understanding of the process, THM in healthcare professionals remains low. This may be explained by the inability to take time out to wash hands because the professionals must maintain peak performance in an emergency that is running at full speed. On the other hand, the main reasons for the low THM are the non-recognition of the need for washing before and after contact with the patient as well as the organizational issue, including that of the equipment. It appears that the implementation of a Lab-C in an emergency room is a good educational tool and a good modulating agent of change that allows the analysis and understanding of the systemic problems too often found in our health establishments. A technological tool is proposed that acts as a stimulus to promote the action of hand washing when the professional enters or leaves the patient's bubble. Additionally, patients could stimulate action by contributing to the audit process.

Le processus d'hygiène des mains à l'urgence est une faille reconnue comme étant une source d'infections nosocomiales associées à la morbidité et la mortalité des patients. Malgré la mobilisation des organismes de santé pour résoudre le problème, le taux d'hygiène des mains (THM) chez les professionnels de la santé demeure insuffisant. Une analyse systémique du processus, en tenant compte de l'historique et du contexte socioculturel des établissements de santé, peut sans doute identifier les motifs susceptibles de freiner le sain processus d'hygiène des mains. Par conséquent, un laboratoire du changement (Lab-C) fut instauré dans une urgence d'un établissement du CHU de Québec en conformité avec les principes inhérents à la théorie historico-socioculturelle de l'activité (CHAT). Un sondage sur le processus d'hygiène des mains fut également réalisé auprès des professionnels de la santé afin de saisir leur compréhension du processus d'hygiène des mains et dans quelle mesure la pandémie à COVID-19 a influencé leurs habitudes. À la suite de ces analyses, nous constatons que les professionnels de la santé connaissent très bien le processus d'hygiène des mains. Ils disposent d'un niveau d'agentivité élevé leur permettant d'être réceptifs aux analyses et aux changements soutenant la modélisation d'un sain processus d'hygiène des mains. Cependant, malgré ces qualités et cette compréhension juste du processus, le THM demeure faible. Ceci peut s'expliquer notamment par l'impossibilité de prendre une pause pour effectuer le lavage des mains parce que les professionnels doivent maintenir une performance optimale dans une urgence qui fonctionne à plein régime. D'autre part, les principales raisons qui expliquent le faible THM sont la non-reconnaissance du besoin du lavage avant et après contact avec le patient ainsi que l'enjeu organisationnel dont notamment celui du matériel. Il appert que l'implantation d'un Lab-C dans une urgence est un bon outil technopédagogique et un bon agent modulateur du changement qui soutient l'analyse et la compréhension des problèmes systémiques trop souvent retrouvés dans nos établissements de santé. Il est proposé un outil technologique qui agit comme stimulus pour favoriser l'action du lavage des mains lorsque le professionnel pénètre ou sort de la bulle du patient. De plus, le patient peut stimuler l'action en contribuant au processus d'audit. / Hand hygiene process in the emergency department is recognized as a source of nosocomial infections associated with patient morbidity and mortality. Despite the mobilization of health organizations to solve the problem, the hand hygiene rate (THM) among health professionals remains insufficient. A systemic analysis of the process, considering the history and sociocultural context of healthcare facilities, would identify the recurring reasons that may hinder the healthy process of hand hygiene. Therefore, a change laboratory (Lab-C) was set up in an emergency room at the CHU de Québec in accordance with the principles inherent to Cultural Historical Activity Theory (CHAT). A hand hygiene process survey was also conducted among healthcare professionals to better understand the process and how the COVID-19 pandemic has influenced their hand hygiene habits. A hand hygiene process survey was also conducted among healthcare professionals to understand their opinion of the process and how the COVID-19 pandemic has influenced their hand hygiene habits. They have a high level of agency likely to challenge them in the modeling of a healthy hand hygiene process. However, despite these qualities and this correct understanding of the process, THM in healthcare professionals remains low. This may be explained by the inability to take time out to wash hands because the professionals must maintain peak performance in an emergency that is running at full speed. On the other hand, the main reasons for the low THM are the non-recognition of the need for washing before and after contact with the patient as well as the organizational issue, including that of the equipment. It appears that the implementation of a Lab-C in an emergency room is a good educational tool and a good modulating agent of change that allows the analysis and understanding of the systemic problems too often found in our health establishments. A technological tool is proposed that acts as a stimulus to promote the action of hand washing when the professional enters or leaves the patient's bubble. Additionally, patients could stimulate action by contributing to the audit process.

Multiple biomarkers of oxidative stress have been measured and used in human medicine to diagnose and monitor airway disease. The purpose of the study was to determine if similar relationships existed between inflammatory and oxidative stress biomarkers in exhaled breath condensate (EBC), bronchoalveolar lavage fluid (BALF), red blood cells, white blood cells, and plasma; and cytokine expression in airway inflammatory cells and mucosal biopsies of RAO-affected horses and their controls. Sixteen horses in pairs were used: 8 non-RAO-affected (controls) and 8 RAO-affected horses. Samples from all horses were collected at remission (S1), during environmental challenge (S2) and at recovery (S3). RAO-affected horses had significant alterations in cellular glutathione peroxidase (cGPx) activity, ascorbic acid and pH in a number of biological samples. Concentrations of 8-isoprostanes, isofurans, amino acids and mRNA expression of interleukin 4 (IL4), gamma interferon (INFγ), inducible nitric oxide synthase (iNOS), extracellular glutathione peroxidase (GPx-3), and cytosolic superoxide dismutase (SOD-1) were not significantly different or were at the limits of detection. Conductivity was measured and assessed as a potential correctional factor for respiratory fluid dilution. The alterations in biomarker concentrations demonstrate that oxidative stress is an important component of airway inflammation in RAO-affected horses. Further research is warranted in the use of biomarkers and the effects of dietary interventions. / Master of Science

O tabagismo é um problema mundial de saúde pública e é considerado a principal causa de morte evitável no mundo associado com câncer de pulmão, doença pulmonar obstrutiva crônica e infarto agudo do miocárdio. O tabagismo induz alterações morfológicas e funcionais no sistema respiratório. O transporte mucociliar (TMC) é um dos principais mecanismos de defesa do sistema respiratório que pode ser alterado com a fumaça e outros produtos do cigarro. O objetivo desse estudo foi avaliar os efeitos do tabagismo e da cessação do tabagismo no TMC nasal, nas propriedades do muco e sobre marcadores inflamatórios. Trinta e três indivíduos tabagistas foram incluídos no estudo após concordância com o termo de consentimento livre e esclarecido. O recrutamento de voluntários foi realizado na Faculdade de Medicina da Universidade de São Paulo (FMUSP) e no Ambulatório de Cessação do Tabagismo da Disciplina de Pneumologia do Hospital das Clínicas da FMUSP. As variáveis desfecho foram o TMC nasal analisado por meio do teste de trânsito da sacarina, as propriedades do muco por meio do ângulo de contato e da transportabilidade da tosse por alto fluxo e a quantificação de células inflamatórias e concentração de interleucinas (IL)-6 e IL-8 em lavado nasal. Vinte cessadores (idade média: 51 anos, 9 do sexo masculino) foram avaliados no tempo basal do estudo, 1o mês, 3o mês e 12o mês de cessação do tabagismo e 13 tabagistas (média de idade: 52 anos, 6 do sexo masculino) foram avaliados no tempo basal e 12 meses após o basal. As características demográficas, hábito tabágico inicial e morbidades de tabagistas e de cessadores foram similares. No tempo basal do estudo, os tabagistas e cessadores apresentaram disfunção do TMC nasal (17,9 ± 10,1 min e 17,4 ± 7,7 min, respectivamente, p=0,880). A cessação do tabagismo induziu melhora significativa do TMC nasal no 1o mês, 3º mês e 12o mês em 63%, 76% e 85% dos indivíduos, respectivamente. Somente aos 12 meses, foi possível observar melhora na transportabilidade do muco por alto fluxo (~ 23%), porém com aumento do número de macrófagos (2x) em lavado nasal. Não observamos alterações no ângulo de contato do muco e nas concentrações de citocinas em lavado nasal. Concluímos que a cessação do tabagismo induz melhora rápida no TMC nasal, porém melhora nas propriedades do muco foi observada somente após 12 meses de cessação do tabagismo / Smoking is a health problem in the world. It is considered a main cause of preventable death and is associated with lung cancer, chronic obstructive pulmonary disease and myocardium infarction. Smoking induces morphological and functional changes in the respiratory system. Mucociliary clearance (MCC) is one of the main defense mechanisms of the respiratory system that can be affected by smoke and other cigarette products. The aim of this study was to assess the effects of smoking and smoking cessation on nasal MCC, mucus properties and inflammatory biomarkers. Thirty three smokers were included in this study after agreement with the written informed consent. Subject´s recruitment was performed at Faculdade de Medicina da Universidade de São Paulo (FMUSP) and Ambulatório de Cessação do Tabagismo da Disciplina de Pneumologia do Hospital das Clínicas da FMUSP. The outcome variables were nasal MCC evaluated by saccharin transit test, mucus properties using contact angle and mucus transportability by high airflow and quantification of inflammatory cells number and interleukin (IL)-6 and IL-8 in the nasal lavage fluid. Twenty volunteers in the smoking cessation program (mean age: 51 years, 9 male) were assessed at baseline, month 1, month 3 and month 12 after of the smoking cessation and 13 smokers (mean age: 52 years, 6 male) were assessed at baseline and 12 months after baseline. Demographic characteristics, smoking history and morbidities were similar between the two groups. At baseline, smokers showed impaired nasal MCC (17.9 ± 10.1 min and 17.4 ± 7.7 min, respectively, p=0.880). Smoking cessation significantly improved nasal MCC at 1 month, 3 months and 12 months in 63%, 76% and 85% of the subjects, respectively. Only after 12 months of smoking cessation, improvement in mucus transportability by high airflow (~ 23%) was observed, however, with increased number of macrophages (2-fold) in nasal lavage fluid. No changes were observed in mucus contact angle and cytokines concentrations in nasal lavage fluid. We concluded that smoking cessation induces rapid improvement in nasal MCC, however, improvement in mucus properties were observed only after 12 months of smoking cessation

Este trabalho teve por objetivo avaliar as alterações respiratórias e neurológicas resultantes da infecção por via intranasal das diferentes estirpes nacionais do herpesvírus equino tipo 1 (EHV-1) em hamsters comparando sua susceptibilidade com estudos sobre infecção do EHV-1 em camundongos e equinos. Para isso, hamsters sírios machos, três semanas de idade, foram infectados por via intranasal com as estirpes do EHV-1 obtidas a partir de fetos abortados e potro neonato infectados (A4/72, A9/92, A3/97 e Iso/72). Os animais foram pesados e examinados diariamente em busca de sinais clínicos e neurológicos. Conforme os sintomas neurológicos apareceram, grupos de cinco hamsters foram eutanasiados por overdose de isoflurano na primeira etapa do projeto, e, de cetamina/xilazina por via intraperitoneal na segunda etapa. Na necropsia, sistema nervoso central (SNC), pulmão, fígado, baço e timo foram coletados para isolamento viral em cultura de células E-dermal e para análise histopatológica. Na segunda etapa do projeto, a lavagem broncoalveolar (LBA) com 3 ml de PBS por hamsters foi realizada para determinar a resposta inflamatória pulmonar através da contagem total e diferencial de leucócitos. Similar aos experimentos com camundongos, hamsters desafiados com as estirpes virais A4/72 e A9/92 apresentaram manifestações clínicas severas no terceiro dia pós-inoculação (dpi), tais como perda de peso aguda, dispnéia, desidratação, decúbito e morte. Observou-se também sinais neurológicos como hiperexcitabilidade, paralisia, espasmos, perda de propriocepção, andar em círculos e convulsões. Ao contrário dos camundongos, que não desenvolveram a doença; hamsters inoculados com as estirpes virais A3/97 e Iso/72 manifestaram sintomas respiratórios e neurológicos agudos entre quarto e quinto dpi, sendo as alterações respiratórias as mais evidentes, principalmente hemoptise e conjuntivite. O isolamento do vírus a partir do SNC foi positivo em todos os animais infectados; no entanto, todas as amostras de pulmões foram positivas apenas nos grupos infectado pelas estirpes virais A9/92 e A4/72. Todas as estirpes do EHV-1 foram isoladas a partir do baço, no entanto, a partir do timo foram isoladas apenas as estirpes A9/92 e A4/72, e, de fígado somente a estirpe viral Iso/72 não foi isolada. No LBA, a contagem total de leucócitos não demonstrou diferenças de valores significativas entre os grupos experimentais, sendo apenas evidente a presença de eritrócitos, macrófagos e neutrófilos na maioria dos esfregaços dos hamsters infectados. Porém, na contagem diferencial de leucócitos observou-se um aumento significativo de neutrófilo com consequente diminuição significativa de macrófagos nos hamsters infectados pelas estirpes virais A3/97, Iso/72 e A4/72 quando comparados ao grupo controle. Os hamsters infectados pela estirpe viral A9/92 mantiveram valores próximos aos do grupo controle. Ao avaliar microscopicamente o SNC, fígado e pulmão dos hamsters infectados foi observado infiltrado inflamatório, necrose e manguito perivascular em SNC; leucocitose e necrose no parênquima hepático com discreta pericolangite e proliferação de ducto biliar; e em pulmão observou inflamação, necrose, edema e hemorragia em bronquíolos e alvéolos. Concluindo, os resultados apontaram o hamster como a espécie mais susceptível à infecção pelo EHV-1 servindo como um modelo experimental complementar para estudos de doenças respiratória e neurológica provocadas por este agente em equinos / This study aimed to evaluate the respiratory and neurological disorders resulting from intranasal infection with different national strains of equine herpesvirus type 1 (EHV-1) in hamsters comparing their susceptibility to studies about EHV-1 infection in mice and horses. Therefore, male Syrian hamsters, three weeks of age, were infected via intransal with EHV-1 strains obtained from aborted fetuses and neonatal foal infected (A4/72, A9/92, A3/97 and Iso/72). The animals were weighed and examined daily for clinical search and neurological signs. As neurological symptoms appeared five hamsters groups were euthanized by isoflurane overdose in the first stage of the project, and ketamine/xylazine via intraperitoneal in the second stage. At necropsy, central nervous system (CNS), lung, liver, spleen and thymus were collected for virus isolation in E-dermal cell culture and histopathological analysis. In the second stage of the project, bronchoalveolar lavage (BAL) with 3 ml of PBS per hamsters was performed to determine the pulmonary inflammatory response through the total and differential leukocyte count. Similar to the experiments with mice, hamsters challenged with viral strains A4/72 and A9/92 had severe clinical manifestations in the third day post-inoculation (dpi), such as acute weight loss, dyspnea, dehydration, recumbency, and death. It was also observed neurological signs such as hyperexcitability, paralysis, spasms, loss of proprioception, circling and convulsions. Unlike mice, which did not develop disease; hamsters inoculated with viral strains A3/97 and Iso/72 showed acute respiratory and neurological symptoms between fourth and fifth dpi, and the most evident respiratory distress was hemoptysis and conjunctivitis. The virus isolation from CNS was positive in all infected animals; however, all lung samples were positive only in groups infected by A9/92 and A4/72 viral strains. All EHV-1 strains were recovered from spleen, however, only A9/92 and A4/72 viral strains were recovered from thymus, but only Iso/72 viral strain was not recovered from liver. BAL total leukocyte count showed no significant differences in values between the experimental groups but the presence of erythrocytes, macrophages and neutrophils were evident in most smears of infected hamsters. However, in the leukocyte differential count it was observed a significant increase in neutrophils with consequent significant reduction of macrophages in hamsters infected by viral strains A3/97, Iso/72 and A9/92 when compared with the control group. Hamsters infected with the A9/92 viral strain maintained values similar to the control group. When evaluating microscopically the CNS, liver and lungs of infected hamsters it was observed inflammatory infiltrate, necrosis and perivascular cuff in CNS; leukocytosis and necrosis in the liver parenchyma with discrete pericholangitis and bile duct proliferation; and inflammation, necrosis, edema and hemorrhage in the bronchioles and alveoli. Concluding, the results showed the hamster as the most susceptible species to EHV-1 infection serving as a complementary experimental model for studies of respiratory and neurological diseases caused by this agent in horses