Thèse de doctorat : Physiologie et physiopathologie des appareils respiratoires : Paris 12 : 2006. / Titre provenant de l'écran-titre. Pagination : 106 f. Bibliogr. f. 81-100.

Airborne disease transmission is a prominent problem facing an increasingly mobilized world. It involves small droplets (bioaerosols) containing pathogens which form in the lungs and are expelled to the environment, where they may persist in the air until inhaled by others. Conceptually, there are two basic approaches to preventing transmission: protect the potential target, or eliminate the source. To this end, the effectiveness of modifying mucus viscoelasticity, through cation exposure, to prevent pathogen transport via bioaerosols was investigated. In vitro models were developed to explore the proposed mechanisms for droplet formation: shear-induced surface-wave instabilities in the airway lining fluid (ALF) of the upper airways; and film formation during the re-opening of collapsed bronchioles in the lower airways. Droplet formation during tidal breathing was shown to be an inhalation process for both upper and lower airway models, and the bifurcation angle of the first bronchi was relevant to the upper airway model. A simulated cough system was also developed and produced the largest number of droplets. COPD sputum viscoelasticity was characterized and its response to cation presence measured: low concentrations of calcium resulted in increased complex modulus and decreased loss tangent (indicating increased fluid stiffness resulting from higher elasticity). Higher concentrations of calcium had the reverse effect. Using the cough system, calcium treated (low concentration) and untreated sputum were compared: treated sputum produced fewer droplets. Droplet concentration (number per liter of air) correlated well with the magnitude of the complex modulus. Once the reduction in total droplets was established, pathogen transport experiments, in which human rhinovirus (HRV) was added to calcium-treated and untreated COPD sputum, were performed. Cell culture media was exposed to cough-air from the samples and then placed on HRV-sensitized HeLa cells, which were then monitored for cell death. Cell death was observed for untreated sputum samples, but not for cation-treated samples, indicating that reducing bioaerosol formation (through cationic modification of mucus viscoelasticity) prevented airborne transport of the virus. / Engineering and Applied Sciences

Made available in DSpace on 2014-06-11T19:30:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-06-15Bitstream added on 2014-06-13T20:00:45Z : No. of bitstreams: 1 teixeira_pc_dr_jabo.pdf: 1193730 bytes, checksum: 293b8714f4d153a9f87c8fa55a7e9e3d (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / O objetivo deste estudo foi avaliar a resposta fisiológica de estresse no clímax e logo após a metamorfose de rã-touro (Lithobates catesbeianus) em diferentes densidades e triagens aplicadas sendo conduzidos no laboratório e no campo. No experimento Densidade foram testadas as densidade de pré-engorda: 70 animais m-2 (D70), 100 animais m-2 (D100) controle, 150 animais m-2 (D150) e 200 animais m-2 (D200), com três réplicas simultâneas. No Experimento Triagem foram testados os manejos de seleção por tamanho (triagem): Sem Triagem (ST), Triagem Parcial a cada quinze dias (TP), Triagem Total a cada quinze dias (TT), com quatro réplicas simultâneas. Foram avaliados o parâmetro hormonal da corticosterona plasmática (Radioimunoensaio - RIE), os parâmetros hematológicos da série vermelha e branca, bem como a morfologia das células cromafins (rim) e glândulas de muco (pele). A corticosterona plasmática respondeu às distintas condições de densidade e triagem aumentando para as densidades D150 e D200 e para as triagens TP e TT ao longo do tempo experimental no laboratório. Porém no campo, a alta densidade e a situação de cativeiro não foram suficientes para provocar estresse nos animais que apresentaram uma diminuição dos níveis de corticosterona aos 30 dias. Na experimentação de Triagem campo os níveis de corticosterona apresentaram menos expressivos se comparado ao laboratório com diminuição aos 30 dias para TT. A série vermelha respondeu aos distintos estresses de densidade e triagem com o aumento do VCM e HCM de forma semelhante, no laboratório. Já no campo este resultado foi menos expressivo A série branca respondeu ao estresse de densidade com aumento de neutrófilos, basófilos, eosinófilos e monócitos, no laboratório e alterações menos expressivas no campo. Porém respondeu ao... / This work aimed evaluate the physiological response of the stress in the climax and immediately after metamorphosis of bullfrogs (Lithobates catesbeianus) submitted to different densities (D) and to selective handling by animal size (S) conducted in laboratory and field. In the Density experiment four different densities for animal were tested: 70 animals.m-2 (D70), 100 animals.m-2 (D100), density control, 150 animals.m-2 (D150) and 200 animals.m-2 (D200), using 3 simultaneous replicates. In the handling experiment three selective handling by animal size were tested: without selective handling (NS), Partial Selective handling each 15 days (PS), Total Selective handling each 15 days (TS), using 4 simultaneous replicates. For the stress response evaluation hormonal parameter for plasmatic corticosterone (radioimmuneassay - RIE) were quantified. In addition, hematological parameters in red and white blood cells, and the morphology of cromafins cells (kidneys) and mucus glands (epiderm) were determined. The plasma corticosterone responded to the different conditions of density and separate by size (screening), increasing to the D150 and D200 and to the PS and TS throughout the experimental period. In the laboratory, but in the field high density and captive situation were not enough to cause stress in animals that showed a decrease in corticosterone levels at 30 days. In the field, levels of corticosterone in screenig stress showed less significance compared to the laboratory with a decrease to 30 days for TS. The red series responded to different stresses, density and screening with the increase MCV and MCH in a similar way in the laboratory, but in the field this result was less significant. The white series responded to density stress with an increase of neutrophils, basophils, eosinophils and monocytes in the... (Complete abstract click electronic access below)

Thesis (Masters Diploma (Medical Technology) -- Cape Technikon, Cape Town, 1992 / Gastric cancer, a fatal malignancy, is endemic in the Coloured population of the Western Cape region of South Africa. Diagnosis is based mainly on histologic investigation with patients of either sex being mainly between 40-60 years of age. The extent to which genetic and environmental influences play a role in the aetiology of the disease is unknown. This study is an attempt to biochemically characterise gastric mucins or mucus glycoproteins, (the main gel forming components of crude mucus scrapings off the mucosal surface), in carcinoma of the stomach (HCA), as compared to those in ulcer disease (HGU), post mortem specimens (PM) and samples obtained from organ transplant donor stomachs (HD). The aim of this study is the development of a diagnostic marker within mucus secretions for the detection of pre-malignant disease amongst the high risk population of the Western Cape region of South Africa. Mucins were extracted from crude mucus gel scrapings according to a carefully designed technique in which proteolytic inhibitors were used to minimise the possibility of endogenous proteolysis in the laboratory through possible contamination. Two density gradient ultra-centrifugation steps for 48 hours each at 105,000g in caesium chloride, a well established standard isolation procedure for mucins, gave a yield of pure mucins which fractionated at a density of approximately 1.41gjml in all groups. These mucins, from the HO, PH, HGU and HCA groups eluted mainly in the included volume of a Sepharose 2B column as broad, polydisperse peaks, suggesting that they were degraded and comprised mainly lower molecular weight PAS positive material in relation to large polymeric gel forming mucin.

Orientador: Carlos Alberto de Magalhães Lopes / Banca: Silvio Luis de Oliveira / Banca: Vidal Haddad Junior / Banca: Mônica Valdyrce dos Anjos Lopes Ferreira / Banca: Itamar Alves Martins / Resumo: Nossos estudos objetivaram caracterizar as respostas imune inata e específica induzidas pelas peçonhas do muco e do ferrão do bagre Cathorops agassizii . A coleta dos espécimes foi realizada no complexo Baía-Estuário de Santos e São Vicente, localizado no litoral sul do Estado de São Paulo. As peçonhas (do Muco e do Ferrão) apresentaram perfil eletroforético similar entre si. Induzida a inflamação aguda em um modelo experimental murino, as peçonhas apresentaram igualmente a capacidade de induzir aumento da permeabilidade vascular e também edema de pata. A detecção de Leucotrieno B4 e Prostaglandina E2 no lavado da cavidade peritoneal dos camundongos injetados, com ambas as peçonhas, corroboram esta hipótese. Nossos resultados através da microscopia intravital mostraram que as peçonhas induzem um grande número de leucócitos rolantes nas vênulas pós-capilares com focos de extravasamento leucocitário, principalmente de neutrófilos seguido pelo influxo de macrófagos. Além disso, a peçonha do Ferrão induziu uma resolução mais rápida do influxo leucocitário ao contrário da peçonha do Muco que manteve o infiltrado macrofágico por até 7 dias. De maneira interessante, somente a citocina IL-6 foi detectada no lavado peritoneal induzida principalmente pela peçonha do Muco e as quimiocinas KC e MCP-1, por ambas as peçonhas, expressando naquele momento, a participação destes mediadores no recrutamento de neutrófilos e macrófagos para o sítio da lesão. As peçonhas foram eficazes ao induzir uma produção primária e secundária de anticorpos das classes IgM e IgG anti-venenos. Observamos ainda, uma especificidade dos anticorpos produzidos para os componentes das próprias peçonhas e também uma reatividade antigênica cruzada entre elas. / Abstract: Our studies aimed to characterize the innate and specific immune responses induced by poisons of the mucus and sting of the catfish Cathorops agassizii. The collection of specimens was accomplished in the complex Bay-Estuary System of Santos and São Vicente located in the south coast of the São Paulo State. Poisons of the Mucus and Sting presented similarities in their electrophorectical profiles. In basis of an Induced process of inflammation throughout an experimental murine model, both poisons equally showed the capacity to increase the vascular permeability and also paw edema. The Leucotrien B4'S and Prostaglandine E2 detection in washings of mice peritoneal cavity injected with both poisons corroborates this hypothesis. Our results through an intravital microscopy procedure also showed that the poisons induced a great number of rolling leukocytes in the post-capillary venules with focus on the leukocyte overflow mainly neutrophiles followed by macrophage influx. Besides, the sting poison Sting induced a faster resolution of the leukocyte influx, as well as, the Mucus poison unlike maintained a macrophage infiltrated for up to 7 days. In an interesting way, only the cytokine IL-6 was detected in peritoneal washings induced mainly by the Mucus poison although Quimiokinins KC and MCP-1 were for both poisons, expressing on that moment the participation of these mediators in the neutrophile and macrophage recruitment for the lesion site. The poisons were also effective in the induction of a primary and secondary production of IgM and IgG classes of antibodies. In this research we still observed a specificity of the antibodies produced for the components of the own poisons and also a crossed antigenic reactivity among them. / Doutor

The mucus coat on the skin and gill of fishes serves a variety of protective functions including lubrication of the body surface, prevention of infection, and deterrence of parasites. Since mucus secretion in some teleosts appears to be controlled by the osmoregulatory hormones, prolactin and Cortisol, it was thought that the mucus coat may participate in teleost hydromineral balance. The primary objective of this study was to -examine the possible role of mucus in-osmoregulation of Leptocottus arma- tus Girard 1854 (Teleostei, Cottidae) and to relate these findings to the -endocrine control of mucus secretion. Whereas Leptocottus skin includes three types of secretory cells (eosinophilic granular, goblet, and cuticle-secreting cells) , the gill epithelium -has only goblet cells. Of ten cottid species examined histochemically, only seawater sculpins produce neutral mucins, while fresh water, seawater, and euryhaline species produce acidic mucins. Leptocottus primarily secretes a sialoglycoprotein, though sulphated mucins are present in gill goblet cells and the cuticle-secreting cells. The granular cells produce a tryptophan-rich basic protein. - Hypophysectomy or seawater adaptation reduce the number of gill goblet cells, compared to respective sham-operated or 5% seawater-acclimated controls. Ovine prolactin treatment of hypophysectomized fish prevented this decrease. In seawater-acclimated Leptocottus prolactin injections increase the number of gill mucus cells, while Cortisol injections of 5% "seawater-adapted fish had the opposite effect. The cuticle-secreting cells were most active in 5% seawater-adapted fish and this state was maintained by prolactin. A moderately active state was typical of seawater-adapted fish and this condition could be maintained by Cortisol or ovine growth hormone. The lowest activity of the cuticle-secreting cells occurred in hypophysectomized fish. Though prolactin and Cortisol appear to control the gill goblet cells and the cuticle-secreting cells, neither hy-pophysectomy nor treatment with prolactin or Cortisol significantly affected the skin goblet cells. The mucus coat appears incapable of reducing integumental permeability through steric interference of diffusion or through Donnan exclusion of ions from the mucus coat. Further, it is argued that enhancement of unstirred layers by mucus could not significantly affect hydromineral balance. Instead, alterations of the mucus coat with environmental salinity may be associated with the lubricating properties of mucus. Leptocottus mucus is a viscous non-newtonian fluid when concentrated but is an efficient lubricant when dilute. Seawater-adapted Leptocottus produce a more efficient lubricating mucus than do 5% seawater-acclimated fish; this effect appears to involve the cuticle-secreting cells. In an associated study I showed that the skin of the goby Gillichthys mirabilis participates actively in osmoregulation through extra-renal ion excretion. Thus the effects of prolactin and Cortisol may be primarily to control active transport in the skin. / Science, Faculty of / Zoology, Department of / Graduate

Cystic fibrosis (CF) is a common lethal hereditary disease resulting from mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR dysfunction affects multiple organ systems and most morbidity and mortality in CF results from lung disease. The CF lung appears healthy at birth, but spontaneously develops airway disease characterized by infection, inflammation, mucus plugging and airway remodeling. A CF pig model was recently generated to determine the events that initiate lung disease. CF pigs recapitulate many findings seen in humans with CF, including the spontaneous development of lung disease. I used newborn CF pigs to investigate two leading hypotheses regarding CF disease initiation: abnormal airway surface liquid (ASL) composition and defective mucociliary transport (MCT). I developed an assay to study ASL composition and found that CF ASL contained similar sodium concentrations, elevated potassium concentrations, and a decreased fraction of volatile material. I developed an assay to measure MCT in vivo. By tracking individual particles in 3-dimensions I found that newborn pigs exhibit a ventrally directed cilia orientation in the trachea. I also found that MCT is highly heterogeneous and particles traveled at different speeds within airways and between airways, challenging the classic view that airway mucus exists as continuous blanket. Comparing particle transport revealed that non-CF and CF newborn pigs exhibit similar basal particle clearance and speeds. Cholinergic stimulation induces mucus and fluid secretion. Particles became stuck in newborn CF pigs after cholinergic stimulation and stasis persisted with tissue submersion. This challenged the leading hypothesis that attributes CF airway disease pathogenesis to ASL depletion. I hypothesized that adherent mucus impairs mucociliary transport in CF airways and I developed an assay to visualize mucus stasis in submerged tracheal segments ex vivo. CF trachea stimulated in vivo exhibited highly adhesive mucus entities that emerged exclusively from submucosal gland ducts. These adherent entities impaired MCT even with extremely high ASL depths. Non-CF trachea with combinatorial disruption of HCO3- and Cl- transport reproduced the defect in CF signifying that anion transport disruption was responsible for adherent mucus. These data suggest that CFTR disruption directly produces multiple host defense defects, including defective bacterial killing and abnormally adherent mucus. Therapeutic targeting of the described defects may provide new opportunities to intervene early and improve the lives of those with CF.

An improved understanding of the mucus biophysical alteration in chronic obstructive pulmonary disease (COPD) and cystic fibrosis (CF) patients is a milestone towards a more accurate and effective treatment of these common and highly debilitating conditions. Little is known about the variations in mucus structure between ill and healthy individuals. An established fact, however, is the role of the mucus obstruction in the exacerbation of the two conditions which impairs the physiological cleaning mechanism of the airways (mucociliary clearance) and is assessed as the strongest predictor of mortality. Therefore, it appears relevant to investigate which properties and structural changes are responsible of the impaired clearance of airway mucus. This thesis presents investigations of mucus on the basis of mucin structure observed in healthy, COPD and cystic fibrosis bronchial mucus samples. AFM (atomic force microscopy) and synchrotron SAXS (small angle X-ray scattering) techniques were used to characterise the structural features of the mucin molecules and allowed to identify the dumbbell structure of airways mucin monomers. The analysis of structural and dimensional features of mucins, highlighted a greater similarity of COPD with the healthy sample rather than with cystic fibrosis. The water sorption analysis using QCM-D (quartz crystal microbalance with dissipation monitoring), established a divergent behaviour between COPD and cystic fibrosis. Compared to healthy specimen, the mucus from COPD donors, showed a greater tendency to absorb water while cystic fibrosis mucus, in contrast, displayed the lowest water absorption.

A microcomputer system called ovulocomputer is proposed. The system is used to predict and detect ovulation of women. It will use image processing and analysis, pattern recognition theory, spectrophotometry, ionometry and conductometry to measure various physical and chemical properties of cervical mucus that are related to blood hormone levels. The functions and characteristics of cervical mucus are introduced in this thesis. Among them, the identification of cervical mucus images, named the ferning test, and related features, are well described. A new idea, called ferning test automation, which has been neither proposed nor developed before, is presented and explained in detail in this thesis.

According to USDA-FSIS reports, L. monocytogenes and Salmonella are two important foodborne pathogens that are prevalent throughout catfish environment. Channel catfish (Ictalurus punctatus) is the most important aquaculture species in the United States accounting for more than 60% of its aquaculture production. The objectives of this study were to determine the effect of strain, temperature concentration on the growth, survival and biofilm formation of L. monocytogenes and Salmonella using catfish mucus extract on different food-contact surfaces. Growth and survival of L. monocytogenes and Salmonella was greater at the higher concentration of mucus extract at both 10°C and 22°C. In 15 micrograms/ml catfish mucus extract L. monocytogenes and Salmonella counts increased to 4.5 log CFU/ml after 72 h at 10°C and 5-7 log CFU/ml counts after 32-48 h at 22°C. In 375 micrograms/ml catfish mucus extract L. monocytogenes and Salmonella counts increased to 6-7 log CFU/ml counts after 72 h at 10°C and 8-9 log CFU/ml counts after 32-48 h at 22°C. L. monocytogenes and Salmonella were able to grow and survive for more than 63 days with at least 4-6 log CFU/ml at 10°C and 6-8 log CFU/ml in 375 micrograms/ml and 15 micrograms/ml of catfish mucus extract respectively. No differences (P > 0.05) among L. monocytogenes and Salmonella strains were observed to form biofilms in the presence catfish mucus extract on the stainless steel surface. The biofilm formation by L. monocytogenes and Salmonella in catfish mucus extract was less (P < 0.05) on buna-n rubber when compared to stainless steel, polyethylene and polyurethane surfaces. Therefore, the findings in this study show that catfish mucus promotes L. monocytogenes and Salmonella to grow and subsequently form biofilms on different food-contact surfaces also promotes growth and survival of these pathogens for longer period of time in catfish processing industries.