Transportadores de iones en pulmón: Uso como dianas terapéuticas / Ion transporters in the lungs. Use as therapeutic targets

Los canales de cloruros, de sodio, de bicarbonato y los de agua (aquaporinas) se coordinan para mantener la cubierta líquido superficial de las vías respiratorias, que es necesaria para el aclaramiento mucociliar. El mecanismo general para el transporte de electrolitos y agua depende principalmente de la expresión diferencial y distribución de los transportadores y bombas de iones. Los iones y el agua se mueven a través de las vía paracelular o transcelular. La ruta transcelular del transporte de electrolitos requiere un transporte activo (dependiente de ATP) o pasivo (siguiendo gradientes electroquímicos) de iones. La ruta paracelular es un proceso pasivo que está controlado, en última instancia, por los gradientes electroquímicos transepiteliales predominantes. La fibrosis quística es una enfermedad hereditaria que se produce por mutaciones en el gen que codifica la proteína reguladora de la conductibilidad transmembrana de la fibrosis quística (CFTR) que actúa como un canal de cloro y cumple funciones de hidratación del líquido periciliar y mantenimiento del pH luminal. La disfunción del canal de cloro en el epitelio respiratorio determina una alteración en las secreciones bronquiales, con aumento de su viscosidad y alteración de la depuración mucociliar y que asociado a procesos infecciosos puede conducir a daño pulmonar irreversible. La disfunción del CFTR, también se ha visto implicado en la patogénesis de la pancreatitis aguda, en la enfermedad pulmonar obstructiva crónica y la hiperreactividad en el asma. Existen fármacos que aprovechan los mecanismos fisiológicos en el transporte de iones, con un objetivo terapéutico.

The chloride channels, sodium and bicarbonate channels, and aquaporin water channels are coordinated to maintain the airway surface liquid that is necessary for mucociliary clearance. The general mechanism for the transport of electrolytes and fluids depends mainly on the differential expression and distribution of ion transporters and pumps. Ions and water move through the paracellular or transcellular pathways. The transcellular route of electrolyte transport requires an active transport (dependent on ATP) or passive (following electrochemical gradients) of ions. The paracellular pathway is a passive process that is ultimately controlled by the predominant transepithelial electrochemical gradients. Cystic fibrosis is a hereditary disease that is produced by mutations in the gene that encode cystic fibrosis transmembrane conductance regulatory protein (CFTR) that acts as a chloride channel and performs functions of hydration of periciliary fluid and maintenance of luminal pH. The dysfunction of the chlorine channel in the respiratory epithelium determines an alteration in the bronchial secretions, with an increase in its viscosity and alteration of the mucociliary clearance and that associated with infectious processes can lead to irreversible lung damage. CFTR dysfunction has also been implicated in the pathogenesis of acute pancreatitis, chronic obstructive pulmonary disease, and bronchial hyperreactivity in asthma. There are drugs that exploit physiological mechanisms in the transport of ions with a therapeutic objective.

Nasal mucociliary transportability of male and female smokers / Transportabilidade mucociliar nasal de homens e mulheres tabagistas

Abstract Introduction: Female smoker's present increased susceptibility to several diseases when compared to the opposite gender. However, there are no studies showing differences in nasal mucociliary transport behavior between male and female smokers. Objective: To compare the nasal mucociliary transportability in male and female smokers and non-smokers, taking into consideration age, anthropometric data, smoking load and pulmonary function. Methods: The analysis included 139 individuals (33 men and 37 women smokers and 32 men and 37 women non-smokers). All participants answered an initial interview to obtain personal data and smoking load. Anthropometric data and carbon monoxide in the exhaled air were assessed. Individuals also performed pulmonary function test and Saccharin Transit Time test. To compare saccharin transit time values between men and women, smokers and non-smokers, stratification of all independent variables was performed (sociodemographic, smoking and respiratory variables) into two categories: below and above the median values. Results: There was no difference between men and women, smokers and non-smokers, regarding nasal mucociliary transportability. Significant differences were only observed between non-smokers. Among those with less forced vital capacity values (<97.37% of predicted), women presented mucociliary transport faster than men. Moreover, it was observed influence of BMI and COex (women smokers), FCV and FEV1 (men non-smokers) and FEF25-75% (women non-smokers) on saccharin transit time values. Conclusion: Based on the findings of this study, nasal mucociliary transport in male and female adult smokers, apparently healthy, are similar.

Resumo Introdução: Mulheres tabagistas apresentam maior susceptibilidade à diversas doenças quando comparadas ao sexo masculino. No entanto, não há estudos mostrando diferenças no comportamento do transporte mucociliar nasal entre tabagistas do sexo masculino e feminino. Objetivo: Comparar a transportabilidade mucociliar nasal em homens e mulheres fumantes e não fumantes, levando em consideração idade, dados antropométricos, carga tabágica e função pulmonar. Método: A análise incluiu 139 indivíduos (33 homens e 37 mulheres fumantes e 32 homens e 37 mulheres não fumantes). Todos os participantes responderam a uma entrevista inicial para a obtenção de dados pessoais e a carga tabágica. Dados antropométricos e monóxido de carbono no ar expirado foram avaliados. Os indivíduos também fizeram teste de função pulmonar e o teste de trânsito de sacarina. Para comparar os valores do teste de trânsito de sacarina entre homens e mulheres, fumantes e não fumantes, foi feita a estratificação de todas as variáveis independentes (variáveis sociodemográficas, tabágicas e respiratórias) em duas categorias: abaixo e acima dos valores médios. Resultados: Não houve diferenças entre homens e mulheres, fumantes e não fumantes, em relação à transportabilidade mucociliar nasal. Diferenças significativas foram observadas apenas entre não fumantes. Entre os que apresentaram valores menores de capacidade vital forçada (< 97,37% do previsto), as mulheres apresentaram transporte mucociliar mais rápido do que os homens. Além disso, observou-se influência do IMC e COex (mulheres fumantes), CVF e VEF1 (homens não fumantes) e FEF25%-75% (mulheres não fumantes) sobre os valores do teste de trânsito de sacarina. Conclusão: Com base nos achados deste estudo, o transporte mucociliar nasal em tabagistas masculinos e femininos adultos, aparentemente saudáveis, são semelhantes.

Effects of passive inhalation of cigarette smoke on structural and functional parameters in the respiratory system of guinea pigs / Efeitos da inalação passiva da fumaça de cigarro em parâmetros estruturais e funcionais no sistema respiratório de cobaias

ABSTRACT Objective: To evaluate the effects of passive inhalation of cigarette smoke on the respiratory system of guinea pigs. Methods: Male guinea pigs were divided into two groups: control and passive smoking, the latter being exposed to the smoke of ten cigarettes for 20 min in the morning, afternoon and evening (30 cigarettes/day) for five days. After that period, inflammatory parameters were studied by quantifying mesenteric mast cell degranulation, as well as oxidative stress, in BAL fluid. In addition, we determined MIP, MEP, and mucociliary transport (in vivo), as well as tracheal contractility response (in vitro). Results: In comparison with the control group, the passive smoking group showed a significant increase in mast cell degranulation (19.75 ± 3.77% vs. 42.53 ± 0.42%; p < 0.001) and in the levels of reduced glutathione (293.9 ± 19.21 vs. 723.7 ± 67.43 nM/g of tissue; p < 0.05); as well as a significant reduction in mucociliary clearance (p < 0.05), which caused significant changes in pulmonary function (in MIP and MEP; p < 0.05 for both) and airway hyperreactivity. Conclusions: Passive inhalation of cigarette smoke caused significant increases in mast cell degranulation and oxidative stress. This inflammatory process seems to influence the decrease in mucociliary transport and to cause changes in pulmonary function, leading to tracheal hyperreactivity.

RESUMO Objetivo: Avaliar os efeitos da inalação passiva da fumaça de cigarro no sistema respiratório de cobaias. Métodos: Foram utilizadas cobaias machos, divididas em dois grupos: controle e tabagismo passivo, esse último exposto à fumaça de dez cigarros por 20 min pela manhã, tarde e noite (30 cigarros/dia) por 5 dias. Após esse período, parâmetros inflamatórios foram estudados através da contagem de degranulação de mastócitos no mesentério e de estresse oxidativo a partir do LBA. Adicionalmente, foram verificadas PImáx, PEmáx, transporte mucociliar (in vivo) e contratilidade traqueal (in vitro). Resultados: Na comparação com o grupo controle, o grupo tabagismo passivo apresentou um aumento significativo na degranulação de mastócitos (19,75 ± 3,77% vs. 42,53 ± 0,42%; p < 0,001), nos níveis de glutationa reduzida (293,9 ± 19,21 vs. 723,7 ± 67,43 nM/g de tecido; p < 0,05) e uma redução significativa no transporte mucociliar (p < 0,05), provocando alterações significativas na função pulmonar, tanto na PImáx como na PEmáx (p < 0,05 para ambas), e hiper-reatividade nas vias aéreas. Conclusões: A inalação passiva da fumaça de cigarro ocasionou aumentos significativos na degranulação de mastócitos e no estresse oxidativo. Esse processo inflamatório parece ter influenciado a diminuição do transporte mucociliar e causado alterações na função pulmonar, proporcionando um quadro de hiper-reatividade traqueal.

Dinámica y patología del barrido mucociliar como mecanismo defensivo del pulmón y alternativas farmacológicas de tratamiento / Mucociliary clearance system in lung defense

The first line of defense to maintain the sterility of the intrapulmonary airways is the mucociliary clearance system. This rapidly responding, non-specific, immune defensive system is challenged by approximately 12,000 liters of air filtered by the lungs each day, containing millions of particles like spores, environmental and eventually toxic pollutants. The main components of this system are the floating mucus "blanket", the underlying mucus propelling cilia with periciliary fluid, and the airway epithelium with secretory and ciliated type of cells. The fine and coordinated regulation of these three components is critical for an effective performance. Deregulation resulting from continuous insults, inherited ion channel disease, infections or abnormal immune response may lead to mucus alterations, goblet cell hyperplasia and metaplasia, airway obstruction, air trapping and chronic lung disease. Options for treatment are limited. Understanding this system may reveal new targets for treatment of lung disease.

The effects on mucociliary clearance of prednisone associated with bronchial section

OBJECTIVE: Infections have been and remain the major cause of morbidity and mortality after lung transplantation. Because mucociliary clearance plays an important role in human defense mechanisms, the influence of drugs on the mucociliary epithelium of patients undergoing lung transplantation must be examined. Prednisone is the most important corticosteroid used after lung transplantation. The aim of this study was to evaluate the effects of bronchial transection and prednisone therapy on mucociliary clearance. METHODS: A total of 120 rats were assigned to 4 groups according to surgical procedure or drug therapy: prednisone therapy (1.25 mg/kg/day); bronchial section and anastomosis + prednisone therapy (1.25 mg/kg/day); bronchial section + saline solution (2 ml/day); and saline solution (2 ml/day). After 7, 15, or 30 days, the animals were sacrificed, and the lungs were removed from the thoracic cavity. The in situ mucociliary transport velocity, ciliary beat frequency and in vitro mucus transportability were evaluated. RESULTS: Animals undergoing bronchial section surgery and anastomosis had a significant decrease in the ciliary beat frequency and mucociliary transport velocity 7 and 15 days after surgery (p<0.001). These parameters were normalized 30 days after the surgical procedure. Prednisone improved mucous transportability in the animals undergoing bronchial section and anastomosis at 15 and 30 days (p<0.05). CONCLUSION: Bronchial section and anastomosis decrease mucociliary clearance in the early postoperative period. Prednisone therapy improves mucus transportability in animals undergoing bronchial section and anastomosis.

Adesividade e purulência de secreções respiratórias: implicações no transporte mucociliar em pacientes com bronquiectasias / Adhesiveness and purulence of respiratory secretions: implications for mucociliary transport in patients with bronchiectasis

OBJETIVO: Analisar e comparar as propriedades de transporte de secreções respiratórias, classificadas através de parâmetros selecionados, de indivíduos com bronquiectasias não secundárias à fibrose cística. MÉTODOS: Foram avaliadas amostras de muco respiratório, classificadas como com propriedades de superfície adesivas ou não adesivas, assim como com aspecto mucoide ou purulento, de 35 participantes com bronquiectasias não secundárias à fibrose cística, quanto a velocidade relativa de transporte (VRT), deslocamento em máquina simuladora de tosse (MST) e ângulo de contato (AC). Para as comparações propostas, foram utilizados modelos de ANOVA, com nível de significância estabelecido em 5 por cento. RESULTADOS: Houve uma diminuição significativa no deslocamento em MST, assim como um aumento significativo no AC, das amostras adesivas quando comparadas às não adesivas (6,52 ± 1,88 cm vs. 8,93 ± 2,81 cm e 27,08 ± 6,13º vs. 22,53 ± 5,92º, respectivamente; p < 0,05 para ambos). O mesmo ocorreu na comparação entre as amostras purulentas e mucoides (7,57 ± 0,22 cm vs. 9,04 ± 2,48 cm e 25,61 ± 6,12º vs. 21,71 ± 5,89º; p < 0,05 para ambos). Não houve diferença na VRT entre os grupos, embora os valores estivessem diminuídos, independentemente da adesividade (adesivas: 0,81 ± 0,20; não adesivas: 0,68 ± 0,24) ou do aspecto (purulentas: 0,74 ± 0,22; mucoides: 0,82 ± 0,22) das amostras. CONCLUSÕES: A secreção respiratória de pacientes com bronquiectasia apresentou uma diminuição do transporte ciliar. Maior adesividade e purulência favorecem a piora das propriedades de transporte, demonstradas pela diminuição do deslocamento em MST e pelo aumento do AC.

OBJECTIVE: To analyze and compare the transport properties of respiratory secretions, classified by selected parameters, in individuals with bronchiectasis unrelated to cystic fibrosis. METHODS: We collected mucus samples from 35 individuals with bronchiectasis unrelated to cystic fibrosis. The samples were first classified by their surface properties (adhesive or nonadhesive), as well as by their aspect (mucoid or purulent). We then tested the samples regarding relative transport velocity (RTV), displacement in a simulated cough machine (SCM), and contact angle (CA). For the proposed comparisons, we used ANOVA models, with a level of significance set at 5 percent. RESULTS: In comparison with nonadhesive samples, adhesive samples showed significantly less displacement in the SCM, as well as a significantly higher CA (6.52 ± 1.88 cm vs. 8.93 ± 2.81 cm and 27.08 ± 6.13º vs. 22.53 ± 5.92º, respectively; p < 0.05 for both). The same was true in the comparison between purulent and mucoid samples (7.57 ± 0.22 cm vs. 9.04 ± 2.48 cm and 25.61 ± 6.12º vs. 21.71 ± 5.89º; p < 0.05 for both). There were no significant differences in RTV among the groups of samples, although the values were low regardless of the surface properties (adhesive: 0.81 ± 0.20; nonadhesive: 0.68 ± 0.24) or the aspect (purulent: 0.74 ± 0.22; mucoid: CONCLUSIONS: The respiratory secretions of patients with bronchiectasis showed decreased mucociliary transport. Increased adhesiveness and purulence cause the worsening of transport properties, as demonstrated by the lesser displacement in the SCM and the higher CA.

Efeito do óxido nítrico no transporte mucociliar / Effects of nitric oxide in mucociliary transport

As vias aéreas, constituídas por epitélio ciliado e secretor de muco, promovem ao trato respiratório mecanismo de defesa que livra esta superfície das partículas inaladas durante a respiração. É de fundamental importância o entendimento da fisiologia e dos mecanismos envolvidos com a atividade mucociliar. A literatura sugere que o NO, em especial o produzido pela expressão da iNOS, mantém a função mucociliar e a defesa imune da cavidade nasal. Objetivo: Avaliar o envolvimento do NO e das vias enzimáticas da produção do NO no transporte mucociliar, utilizando inibidores da NO sintase constitutiva e indutiva, L-NAME e aminoguanidina, respectivamente. Materiais e métodos: Preparações de palatos de rã foram imersos em soluções de ringer (controle), L-NAME ou aminoguanidina. Os palatos foram imersos nestas soluções por quatro períodos de 15 minutos. Medidas da velocidade do transporte mucociliar foram feitas antes e após cada exposição. Resultos: Palatos controles mantiveram estável a velocidade do transporte. O L-NAME aumentou, enquanto a aminoguanidina reduziu a velocidade de transporte do muco. Conclusão: O bloqueio inespecífico da cNOS com L-NAME e bloqueio relativamente específico da iNOS com aminoguanidina permitiu propor que dependendo da via o NO pode aumentar ou diminuir o transporte mucociliar em palatos de rã.

The airways are made up of ciliated epithelium which secretes mucous, protecting the respiratory tract from particles inhaled during breathing. Its is paramount to understand the physiology and the mechanisms involved in mucociliary activity. Literature suggests that Nitric oxide (NO), especially the one produced by iNOS expression, maintains the mucociliary function and the immune defense of the nasal cavity. AIM: to assess NO participation and the enzymatic pathways in the production of NO and mucociliary transport, using constructive and inductive NO synthetase inhibitors, L-NAME and aminoguanidine, respectively. Materials and methods: frog palates were prepared and immerse in ringer (control), L-NAME or aminoguanidine solutions. The palates were immerse in these solutions for four periods of 15 minutes. Mucociliary transport measures were carried out before and after each exposure. Results: control palates maintained stable their transportation speed. L-NAME increased, while aminoguanidine reduced mucous transportation velocity. Conclusion: unspecific cNOS block with L-NAME and relatively specific iNOS block with aminoguanidine results leads us to propose that depending on the pathway, the NO can increase or reduce mucociliary transport in frog palates.

Influência da técnica de pressão expiratória positiva oscilante utilizando pressões expiratórias pré-determinadas na viscosidade e na transportabilidade do escarro em pacientes com bronquiectasia / Influence that oscillating positive expiratory pressure using predetermined expiratory pressures has on the viscosity and transportability of sputum in patients with bronchiectasis

OBJETIVO: Verificar a efetividade da técnica de pressão expiratória positiva oscilante (PEPO) utilizando pressões expiratórias pré-determinadas sobre a viscosidade e a transportabilidade do escarro em pacientes com bronquiectasia. MÉTODOS: Foram incluídos no estudo 15 pacientes estáveis com bronquiectasia (7 homens; média de idade = 53 ± 16 anos), submetidos a duas intervenções PEPO consecutivas, com 24 h de intervalo entre si, utilizando pressões expiratórias de 15 cmH2O (P15) e 25 cmH2O (P25). O protocolo consistiu de tosse voluntária; nova expectoração voluntária após 20 min, denominado tempo zero (T0); repouso de 10 min; e utilização da técnica em duas séries de 10 min (S1 e S2) de PEPO em P15 e P25, com intervalo de 10 min entre si. A viscosidade e transportabilidade do escarro foram avaliadas pela viscosimetria, velocidade relativa de transporte no palato de rã, deslocamento em máquina simuladora de tosse e ângulo de adesão. As amostras de escarro foram coletadas em T0, após S1 e após S2. Testes estatísticos específicos foram aplicados de acordo com a distribuição dos dados. RESULTADOS: Houve diminuição significante da viscosidade do escarro após S1 em P15 e após S2 em P25. Não houve diferenças significantes entre todas as amostras para a transportabilidade. CONCLUSÕES: Houve diminuição da viscosidade do escarro quando a PEPO foi realizada em P15 e P25, o que sugere que não seja necessário gerar alta pressão expiratória para obter o resultado desejado.

OBJECTIVE: To determine the effectiveness of oscillating positive expiratory pressure (OPEP) using predetermined expiratory pressures on the viscosity and transportability of sputum in patients with bronchiectasis. METHODS: The study involved 15 stable patients with bronchiectasis (7 males; mean age = 53 ± 16 years), submitted to two consecutive OPEP interventions, with a 24-h interval between the two, using positive expiratory pressures set at 15 cmH2O (P15) and 25 cmH2O (P25). The protocol consisted of a voluntary cough; another voluntary cough 20 min later, designated time zero (T0); a 10-min rest period; and two 10-min series (S1 and S2, using OPEP at P15 and P25 in both), with a 10-min interval between the two. The viscosity and transportability of sputum were evaluated by viscometry, relative transport velocity on frog palate, transport in a simulated cough machine and contact angle. Sputum samples were collected at T0, after S1 and after S2. Specific statistical tests were performed depending on the type of data distribution. RESULTS: In comparison with the values obtained at T0, sputum viscosity decreased significantly after S1 at P15 and after S2 at P25. There were no significant differences among all of the samples in terms of transportability. CONCLUSIONS: The fact that sputum viscosity decreased whether OPEP was performed at P15 or at P25 suggests that there is no need to generate high expiratory pressure to achieve the desired result.

Analysis of the sputum and inflammatory alterations of the airways in patients with common variable immunodeficiency and bronchiectasis

INTRODUCTION: Common variable immunodeficiency is characterized by defective antibody production and recurrent pulmonary infections. Intravenous immunoglobulin is the treatment of choice, but the effects of Intravenous immunoglobulin on pulmonary defense mechanisms are poorly understood. OBJECTIVE: The aim of this study was to verify the impact of intravenous immunoglobulin on the physical properties of the sputum and on inflammatory alterations in the airways of patients with Common variable immunodeficiency associated with bronchiectasis. METHOD: The present study analyzed sputum physical properties, exhaled NO, inflammatory cells in the sputum, and IG titers in 7 patients with Common variable immunodeficiency and bronchiectasis with secretion, immediately before and 15 days after Intravenous immunoglobulin. A group of 6 patients with Common variable immunodeficiency and bronchiectasis but no sputum was also studied for comparison of the basal IgG level and blood count. The 13 patients were young (age=36±17 years) and comprised predominantly of females (n=11). RESULTS: Patients with secretion presented significantly decreased IgG and IgM levels. Intravenous immunoglobulin was associated with a significant decrease in exhaled NO (54.7 vs. 40.1 ppb, p<0.05), sputum inflammatory cell counts (28.7 vs. 14.6 cells/mm³, p<0.05), and a significant increase in respiratory mucus transportability by cough (42.5 vs. 65.0 mm, p < 0.05). CONCLUSION: We concluded that immunoglobulin administration in Common variable immunodeficiency patients results in significant improvement in indexes of inflammation of the airways with improvement in the transportability of the respiratory mucus by cough.

Métodos de estudo do transporte mucociliar: [revisão] / Methods for studying mucociliary transport: [review]

Disfunções no transporte mucociliar trazem desde queda na qualidade de vida dos pacientes, como nas rinites e rinossinusites crônicas, até graves conseqüências com risco de seqüelas irreversíveis e mesmo letais, como nos casos de fibrose cística e das discinesias ciliares primárias. Desta forma, torna-se fundamental o conhecimento do funcionamento normal do aparelho mucociliar e de como alterações em seus componentes (cílio, muco-fluido periciliar e interação dinâmica entre ambos) afetam o transporte das secreções respiratórias. OBJETIVOS: Este artigo visa a revisar e discutir as diferentes técnicas de avaliação do transporte mucociliar descrevendo suas peculiaridades e aplicabilidades clínicas e experimentais. CONCLUSÕES: Os métodos citados nesta revisão nos fornecem informações importantes sobre os diferentes aspectos do transporte mucociliar. Alguns apresentam uma maior facilidade de realização e resultados reprodutíveis, já outros apenas mostraram-se com aplicabilidade em protocolos de pesquisa em virtude de dificuldades técnicas e limitações financeiras. Há que se considerar a inexistência de métodos que avaliem ambulatorialmente a freqüência de batimento ciliar (FBC) "in vivo" e "in situ", o que se tornaria uma ferramenta importante, tanto no âmbito científico, quanto na prática clínica, auxiliando no diagnóstico das discinesias ciliares e evitando a realização de procedimentos mais invasivos para a sua confirmação diagnóstica.

Mucociliary transport dysfunctions can impair the quality of life of patients suffering from chronic rhinossinusitis and lead to severe consequences such as alterations in respiratory physiology or even death as in cases of cystic fibrosis and primary ciliary dyskinesia. Therefore, it is crucial to understand the physiology of the mucociliary apparatus and how its components (cilia, mucus-periciliary layer and its interaction) affect the clearance of respiratory secretions. AIMS: This paper aims to review and to discuss different techniques for studying mucociliary transport and their clinical and experimental applicability. CONCLUSIONS: The methods listed in this revision provide us with valuable information about different aspects of the mucociliary transport. Some of the methods listed are more suitable for clinical practice and present reproducible results. Others, show only applicability in experimental settings due to technical difficulties or financial limitations. However, it is important to emphasize that up to now there is no method that can evaluate ciliary beating frequency (CBF) in vivo and in situ. Such a method would become a valuable tool in the scientific scenario and in the clinical practice, supporting the diagnosis of ciliary dyskinesias and avoiding the use of invasive procedures to corroborate the clinical suspicion.

O muco traqueobrônquico humano mantido em temperatura ambiente e suas propriedades físico-químicas / Physicochemical properties of human tracheobronchial sputum maintained at room temperature

OBJETIVO: Verificar a influência da permanência em temperatura ambiente na análise da transportabilidade por ação ciliar e por tosse e do ângulo de contato do muco traqueobrônquico. MÉTODOS: Foi coletado muco hialino de 30 indivíduos sem doença pulmonar, e purulento de vinte pacientes com bronquiectasia. As amostras foram analisadas logo após a coleta e novamente após 24 h. RESULTADOS: Para o muco purulento, após 24 h em temperatura ambiente, houve aumento no deslocamento por tosse (96 ± 50 vs. 118 ± 61 mm) e diminuição do ângulo de contato (32 ± 6 vs. 27 ± 6 graus) (p < 0,05). Para o muco hialino não houve alterações nas medidas analisadas. CONCLUSÃO: O muco traqueobrônquico hialino pode ser armazenado em temperatura ambiente por 24 h sem que haja alterações em sua transportabilidade por ação ciliar ou em seu ângulo de contato. Por outro lado, o muco purulento não deve permanecer em temperatura ambiente por muitas h para que não se altere seu ângulo de contato e sua transportabilidade por tosse.

OBJECTIVE: To evaluate the effect that maintaining tracheobronchial sputum at room temperature has on the analysis of ciliary transport and cough, as well as on the contact angle. METHODS: Hyaline sputum was collected from 30 individuals without pulmonary diseases, and purulent sputum was collected from patients with bronchiectasis. The samples were analyzed immediately after collection and again after 24 h. RESULTS: After 24 h at room temperature, the purulent sputum presented an increase in cough-induced dislodgment (96 ± 50 vs. 118 ± 61 mm) and a decrease in the contact angle (32 ± 6 vs. 27 ± 6 degrees) (p < 0.05). For the hyaline sputum, there were no alterations in the parameters analyzed. CONCLUSION: Hyaline tracheobronchial sputum can be stored in room temperature for 24 h without presenting alterations in ciliary transport or contact angle. However, purulent sputum should not be stored at room temperature for many hours, since ciliary transport and contact angle might be altered as a result.

Ação do estrógeno e progesterona na mucosa nasal humana: avaliação do transporte mucociliar nasal de sacarina e pesquisa de receptores hormonais através de método imuno-histoquímico / Estrogen and progesterone influence in human nasal mucosa: evaluation of nasal saccharin mucociliary transport and test for hormone receptors with immunohistochemical staining

Apesar do estudo exaustivo do transporte mucociliar nasal, ainda há dados controversos sobre a influência direta dos hormônios sexuais femininos nesse mecanismo. O presente estudo teve por objetivos: 1. avaliar o transporte mucociliar nasal de sacarina nos sexos masculino e feminino, comparando-o nas fases folicular, periovulatória e lútea de ciclos ovarianos consecutivos e 2. identificar a expressão e a localização dos receptores para estrógeno e progesterona na mucosa nasal humana em conchas nasais inferiores de indivíduos dos sexos masculino e feminino na idade reprodutiva. O transporte mucociliar nasal de sacarina foi avaliado prospectivamente em 14 voluntários não fumantes, sem queixas nasais, com idades entre 15 e 30 anos (7 homens e 7 mulheres, com média de idade 23,5 anos). Nas mulheres, o transporte mucociliar nasal de sacarina foi medido nas fases folicular, periovulatória e lútea durante dois ciclos ovarianos consecutivos (em cinco casos) ou três ciclos consecutivos (em dois casos). Nos homens, o transporte mucociliar nasal de sacarina foi avaliado em medidas repetidas aleatoriamente três vezes (em dois casos) ou seis vezes (em cinco casos). A expressão dos receptores para estrógeno e progesterona na mucosa nasal humana foi avaliada por método imunohistoquímico, de modo retrospectivo, em conchas nasais inferiores conservadas em formaldeído e fixadas na parafina, arquivadas após a cirurgia de turbinectomia parcial da concha inferior a que foram submetidos 20 pacientes da mesma faixa etária dos voluntários (10 pacientes do sexo masculino e 10 do sexo feminino, com idades entre 15 e 33 anos, média de idade 22,1 anos). Para a imuno-histoquímica utilizaram-se anticorpos monoclonais de camundongo contra receptores para estrógeno (clone 6F11, Novocastra) e para progesterona (clone 16, Novocastra) separadamente. Não houve diferenças significativas no transporte mucociliar nasal de sacarina entre as fases folicular...

Although nasal mucociliary clearance has been thoroughly studied, there is controversial evidence that it is directly influenced by female sex hormones. This study focused on: 1. evaluating saccharin nasal mucociliary transport in both sexes and during the follicular, periovulatory and luteal phases of consecutive ovarian cycles, and 2. identifying the expression and localisation of estrogen and progesterone receptors in human nasal mucosa from inferior turbinates of patients in reproductive age. Saccharin nasal mucociliary transport was prospectively evaluated in 14 nonsmoking healthy volunteers aged 15 to 30 years (7 males and 7 females, mean age 23.5 years) who had no nasal complaints. In females, saccharin nasal mucociliary transport was measured in the follicular, periovulatory and luteal phases during two consecutive ovarian cycles (five cases) or three consecutive cycles (two cases). In males, the saccharin nasal mucociliary transport was randomly repeated three times (two cases) or six times (five cases). Estrogen and progesterone receptor expression in human nasal mucosa was retrospectively assessed by immunohistochemistry in archival, formalin-fixed, paraffin-embedded inferior nasal conchae from 20 patients submitted to partial inferior turbinectomy whose ages were matched to their of the volunteers (10 male and 10 female patients aged 15 to 33 years, mean age 22.1 years). Immunohistochemistry used mouse monoclonal antibodies against estrogen receptor (6F11 clone, Novocastra) and progesterone receptor (16 clone, Novocastra) separately. There were no significant differences in saccharin nasal mucociliary transport among follicular, periovulatory and luteal phases in consecutive ovarian cycles, nor between sexes (p=.08). Even though, considering the first ovarian cycle only, saccharin nasal mucociliary transport was faster during the follicular phase (p=.03). Estrogen and progesterone receptors were found in the cytoplasm of serous...

Nasal mucociliary clearance in adenotonsillar hypertrophy.

The nasal mucociliary clearance time was studied using Andersen saccharin method in 50 normal children and 50 age and sex matched patients of adenotonsillar hypertrophy, which was repeated one month after adenotonsillectomy. The normal mucociliary clearance time in healthy children was found to be 8.55 +/- 2.11 minutes. A significant impairment in nasal mucociliary clearance time was noted in children suffering with adenotonsillar hypertrophy which was 16.97 +/- 3.1 minutes, and early adenotonsillectomy restored the mucociliary clearance to a normal 8.7 +/- 2.14 minutes.

Tiempo de tránsito nasal de sacarina en escolares sanos / Nasal transit time determined with the saccharine test in healthy children

Introducción. La prueba de tránsito nasal de sacarina es un medio simple, no invasivo e indoloro para evaluar la depuración mucociliar nasal. Ha sido empleada en varias condiciones como en la evaluación de los efectos de intervenciónes quirúrgicas y medicamentosas directas en la región aérea nasal y también de aquéllas que sin estar directamente relacionadas alternan la depuración mucociliar nasal. Sin embargo, existe muy poca información acerca de los valores de dicha prueba en la edad pediátrica. El objetivo de este estudio fue determinar los valores de tiempo de tránsito nasal de sacarina en escolares sanos. Material y métodos. Este estudio se realizó en escolares sanos. La sacarina sódica en polvo fue colocada en el cornete inferior y se computó el tiempo transcurrido hasta que los niños relataban la aparición de un sabor dulce en la lengua. El análisis estadístico de los tiempos de tránsito nasal de sacarina, según edad y sexo, se realizó mediante pruebas no paramétricas (ANOVA), chi cuadrada y la regresión lineal. Resultados. Se estudiaron 84 niños, con edades entre 4 y 14 años. El tiempo promedio de tránsito nasal de sacarina fue de 16.7 ñ 9.2 min y se observó una tendencia significativa del tiempo de sacarina a ser mayor con el aumento de la edad y en niñas (P< 0.05). Conclusión. Los valores descritos en el presente estudio se encuentran dentro del límite referido en publicaciones de otros países y contribuyen a aumentar la escasa información disponible aceraca de los valores de tiempo de tránsito nasal de sacarina en escolares sanos

Estudio del transporte mucociliar nasal en sujetos normales en la ciudad de Arequipa

Se realizó un estudio en 100 adultos normales residentes en la ciudad de Arequipa, con el objeto de determinar el tiempo de transporte mucociliar nasal in vivo, haciendo uso del polvo de carbón vegetal y la inspección faringea como método. Se planteó la hipótesis de que existirían alteraciones en el tiempo de transporte mucociliar nasal dependientes de las condiciones climáticas de temperatura y humedad del medio. Los valores encontrados y presentados en cuadros y figuras se comparan con los obtenidos en otros medios más cálidos y húmedos; y son relacionados con algunas variables dentro de los criterios de normalidad, como son la edad, el sexo, la hora en que se realizaron las observaciones así como el ciclo ovulatorio de la mujer. Se determinó un promedio de 9.0105 minutos como tiempo de transporte mucociliar nasal en sujetos normales en Arequipa, con una desviación estándar de 4.3110. El amplio rango en que se ubicaron estos valores se encontró entre los 3 y 22 minutos. Los aclaradores lentos fueron encontrados en una proporción del 5 por ciento. Al comparar los resultados obtenidos con los observados en trabajos similares ya mencionados, se encontró que el promedio del tiempo de transporte mucociliar nasal fue más alto en Arequipa y que la proporción de aclaradores lentos también fué más alta, sin embargo estas diferencias no fueron estadísticamente significativas. Tampoco se demostraron diferencias significativas al relacionar el tiempo de transporte mucociliar nasal con las variables de edad, sexo, ciclo ovulatorio de las mujeres ni con la hora en que se realizaron las observaciones.

Methods for studying respiratory mucus and mucus clearance

We describe some basic procedures for studying the properties of mucus. These techniques can be applied in both clinical and physiological studies to improve the understanding of the mechanisms related to epithelial defense in health and disease. Mucus collection - A major difficulty is the lack of simple and noninvasive methods for collecting normal mucus in sufficient quantity for later analysis. Physical properties of mucus - A. Rheology: Mucus exhibits both solid and liquid properties and the important factor governing the actual behavior is time. The magnetic microrheometer provides an elegant method for measuring rheological properties of microsamples. B. Adhesivity: It characterizes the forces of attraction between an adherent surface and an adhesive system and can be calculated by measuring the contact angle between a mucus drop and a surface. Mucus Transport - A. Mucus transport by cilia: Mucus is primarily cleared by the continuous ciliary beating, which can be studied using techniques such as the frog palate preparation as well as direct measurement. i.e., in situ mucus clearance. B. Cough clearance: It is essential for elimination of secretions in diseases leading to hypersecretory states. The cough machine simulates the flow-time profile of human coughing. Transepithelial potential difference - A potential difference exists between the epithelial surface and the submucosa and is the net result of the activity of the ion-transport system of the pulmonary epithelium. The potential can be measured using appropriate microelectrodes. Quantitative morphology - Methods may be used to characterize the epithelial surface condition that continuously changes during aggressive conditions.

Mecanismos de defensa del aparato respiratorio / Defensive mechanisms of the respiratory tract

El aparato respiratorio debe evitar la agresión y el daño causado por vapores, gases, irritantes, partículas inorgánicas y orgánicas, alérgenos y microorganismos que ingresan con el aire inspirado. Para cumplir esta función posee mecanismos defensivos altamente especializados que comprenden factores mecánicos, reflejos y elementos humorales y celulares. El filtro nasal, la filtración aerodinámica y el transporte mucociliar representan importantes mecanismos de arrera. El estornudo, la tos y la broncoconstricción son los principales reflejos. Entre los factores humorales inespecíficos deben destacarse a lisozima, fibronectina, antiproteasas, factores de complemento y al surfactante alveolar. Además del epitelio ciliado pseudoestratificado, entre los elementos celulares de importancia en los mecanismos de defensa, deben nombrarse a los mastocitos, los polimorfonucleares neutrófilos y eosinófilos, los macrófagos alveolares y los linfocitos. La respuesta inmune con sus características de específicidad, memoria y perfeccionamiento es el nivel más especializado de defensa. En la vertiente humoral destaca la IgA secretoria que cumple funciones de neutralización, bloqueo y opnonización. Finalmente, los linfocitos T ejercen la inmunidad celular mediante citotoxicidad y liberación de linfokinas que activan a células accesorias. Todos los mecanismos descritos deben actuar en forma coordinada y regulada para lograr su objetivo y evitar el daño tisular y la infección. Existen múltiples condiciones que pueden alterar estos mecanismos defensivos y facilitar la acción de los agentes agresores

Effect of steam inhalation on mucociliary activity in patients of chronic pulmonary disease.

Increased mucociliary activity has been observed with several modalities, including inhalation which seems to achieve faster drug delivery. In patients of chronic lung disease, the mucociliary activity is known to be impaired. The effect of steam inhalation on mucociliary transport time was studied in these patients. It was found that steam inhalation improved mucociliary activity significantly (p < 0.001) in both groups receiving either only bronchodilators, or bronchodilators as well as steroids.

High frequency resonant oscillations technique to improve nasal muco-ciliary transport.

A modified high frequency resonant oscillations (HFRO) technique was developed to improve the nasal muco-ciliary transport in patients with impaired ciliary functions due to various chronic diseases of the upper respiratory tract. Twelve healthy males (group I) and ten patients suffering from malignancy of the upper respiratory tract (group II) were exposed to HFRO in order of 200, 250, 300, 400, 500 Hz respectively. The muco-ciliary transport (MCT) was assessed by using saccharine clearance time technique. A significant improvement in MCT was observed in both the groups at all the frequency levels, more so in the range of 400 and 500 Hz. An overall relative improvement of 174 per cent (P less than 0.01) in group I and 145 per cent (P less than 0.001) in group II was observed. No significant side effects were noted.