Citologia dos lavados traqueobrônquico (LTB) e broncoalveolar (LBA) de bezerros holandeses sadios durante o primeiro mês de vida / Cytology of tracheobronchial and bronchoalveolar lavage in healthy Holteins calves during the first month of life

O período neonatal dos bezerros é um momento crítico para adaptação do recém-nascido à vida extra uterina e o sistema respiratório, um dos mais exigidos funcionalmente, é frequentemente afetado por enfermidades, redundando no prejuízo direto da sua função e acarretando perdas econômicas importantes na pecuária. O ponto básico para reduzir estas perdas, é representado pela adequada avaliação clínica dos neonatos, todavia o diagnóstico baseado exclusivamente no exame físico é muito difícil de ser estabelecido. O uso de exames complementares como a citologia do trato respiratório torna-se uma ferramenta diagnóstica importante nestes casos, porém faz-se necessário, padronizar seus achados frente às diferentes técnicas empregadas para a sua obtenção. Assim, o presente estudo propôs-se acompanhar as variações dos constituintes celulares da região traqueobrônquica e broncoalveolar obtidos por lavados respiratórios pelos métodos de traqueocentese e por colheita nasotraqueal respectivamente, durante o primeiro mês de vida de bezerros sadios. Observou-se alteração no quadro citológico ao longo do tempo, quando a região traqueobrônquica foi lavada, expresso por diminuição da porcentagem de macrófagos alveolares, com aumento de neutrófilos, possivelmente, por maior irritação local provocada pela técnica, que se repetiu sequencialmente e/ou por maior estimulo de microorganismos inalados depositados nesta região. Na região broncoalveolar, não encontraram-se variações nos constituintes celulares em função do tempo. Os resultados permitiram a conclusão que a população celular da região traqueobrônquica modificou-se ao longo das semanas de vida dos bezerros, possivelmente pela técnica empregada e/ou fisiologia normal da região, sendo representadas por maiores magnitudes de neutrófilos. De modo diverso, na região broncolaveolar, as células evidenciaram um comportamento estável durante o primeiro mês de vida dos bezerros neonatos, apresentando predomínio numérico dos macrófagos alveolares.

The neonatal calf is a critical moment for adaptation of the newborn to extra uterine life. The respiratory tract is functionally very demanded and often affected by disease, resulting in direct loss of their function and causing serious economic losses in livestock. The basic point to reduce these losses is appropriate clinical evaluation of neonates; but the diagnosis based solely in physical examination is very difficult to establish. The use of complementary analysis such cytology of the respiratory tract becomes an important diagnostic tool; however their findings must be standardized in the face of different techniques employed. This research studied the dynamics of the cellularity of the bronchoalveolar and tracheobronchial region obtained through lung lavage harvested by nasotracheal catheterization technique and tracheocenthesis respectively, during the first month of life of healthy calves. The tracheobronchial cytology was influenced by the time, showing decreased number of alveolar macrophages and greater number of neutrophils, possibly increased by local irritation caused by the technique, which was repeated sequentially, and/or through greater stimulation of inhaled microorganisms deposited in this region. In the bronchoalveolar region no variation in the cellular constituents in function of time was found. The results allowed the conclusion the cell population of the tracheobronchial region has changed over the week-old calves, possibly due to the technique used and/or to the normal region physiology, represented by higher magnitudes of neutrophils. Otherwise, the cells of the broncholaveolar region showed a stable behavior during the first month of life of newborn calves, presenting numerical predominance of alveolar macrophages.

Comparative Quantitative and Qualitative Attributes of the Surface Respiratory Macrophages in the Domestic Duck and the Rabbit / Atributos Cualitativos, Cuantitativos y Comparativos de los Macrófagos de la Superficie Respiratoria en el Pato Doméstico y el Conejo

In mammals, surface respiratory macrophages (SM) are known to play a foremost role in protecting the respiratory system by providing first line of defense through engulfing pathogens and particulate matter respired with air. It has been reported that the pulmonary cellular defense system of domestic birds is inadequate. In particular, low number of SM and even lack of the cells in a healthy avian respiratory system have been associated with susceptibility of domestic birds to respiratory diseases. In an endeavor to resolve the existing controversy, the quantitative and qualitative attributes of the surface respiratory macrophages of the domestic duck and of the domestic rabbit were compared under similar experimental conditions. Quantitatively, the rabbit SM were on average approximately fourteen times more than the duck SM. The SM were found to have comparable diameters measuring about 12 µm in the duck and 13 µm in the rabbit. Similarly, the duck and the rabbit SM were structurally similar. Typically, they were round granular cells possessing filopodial extensions and variable electron dense bodies in the cytoplasm. The phagocytic capacity measured using polystyrene particles revealed that the duck SM had a higher phagocytic capacity than the rabbit SM. The volume density of the engulfed polystyrene particles, i.e. the volume of the particles per unit volume of the cell was estimated at 20 percent in the duck and 9 percent in the rabbit. These results suggest that the comparatively low numbers of SM in domestic birds may contribute to susceptibility of the birds to diseases. However, given the high phagocytic capacity of the avian SM, susceptibility of the domestic birds may not be due to dearth of the SM alone but some other factor (s) such as persistent exposure of the birds to particulate matter which is known to reduce robustness of the SM may be involved.

En los mamíferos, los macrófagos de la superficie respiratoria (SM) son conocidos por jugar el papel más importante en la protección del sistema respiratorio, proporcionando la primera línea de defensa en contra de agentes patógenos y envolviendo las partículas de aire respirado. Se ha informado que el sistema de defensa celular pulmonar de las aves domésticas es insuficiente. En particular, el bajo número de SM, e incluso las células del sistema respiratorio de las aves domésticas, en un ambiente sano, se ha asociado con susceptibilidad a enfermedades respiratorias. En un esfuerzo para resolver la controversia existente, los atributos cuantitativos y cualitativos de los macrófagos de la superficie respiratoria del pato doméstico y el conejo doméstico fueron comparados en las mismas condiciones experimentales. Cuantitativamente, los SM del conejo fueron en promedio aproximadamente catorce veces más que los SM en el pato. Los SM se encontraron con un diámetro comparable, al medir alrededor de 12 micras en el pato y 13 micras en el conejo. Del mismo modo, en el pato y el conejo los SM eran estructuralmente similares. Por lo general, correspondieron a células granulares con extensiones filopodiales y organismos electrodensos variables en el citoplasma. La capacidad fagocítica medida utilizando partículas de poliestireno reveló que los SM del pato tenían una mayor capacidad fagocítica que el conejo. La densidad de volumen de las partículas de poliestireno envueltas, es decir, el volumen de las partículas por unidad de volumen se estimó en 20 por ciento en el pato y 9 por ciento en el conejo. Estos resultados sugieren que el número comparativamente bajo de los SM en las aves domésticas puede contribuir a su susceptibilidad a enfermedades. Sin embargo, dada la alta capacidad fagocítica de los SM aviares, la susceptibilidad de las aves domésticas no puede deberse solamente a la escasez de SM, sino a algunos otros factores pueden estar involucrados, como la exposición persistente de las aves a partículas, las cuales se sabe reducen la robustez de los SM.

The Expression of Adiponectin Receptors and the Effects of Adiponectin and Leptin on Airway Smooth Muscle Cells

PURPOSE: Obesity is a major risk factor for asthma and it influences airway smooth muscle function and responsiveness. Adiponectin is inversely associated with obesity and its action is mediated through at least 2 cell membrane receptors (AdipoR1 and AdipoR2). Leptin is positively associated with obesity. We investigated whether human airway smooth muscle (ASM) cells express adiponectin receptors and whether adiponectin and leptin regulate human ASM cell proliferation and vascular endothelial growth factor (VEGF) release. MATERIALS AND METHODS: Human ASM cells were growth-arrested in serum-deprived medium for 48 hours and then stimulated with PDGF, adiponectin and leptin. After 48 hours of stimulation, proliferation was determined using a cell proliferation ELISA kit. Human AdipoR1 and -R2 mRNA expressions were determined by RT-PCR using human-specific AdipoR1 and -R2 primers. Concentrations of VEGF, monocyte chemotactic protein (MCP)-1 and macrophage inflammatory protein (MIP)-1alpha in cell culture supernatant were determined by ELISA. RESULTS: Both AdipoR1 and AdipoR2 mRNA were expressed in the cultured human ASM cells. However, adiponectin did not suppress PDGF-enhanced ASM cell proliferation, nor did leptin promote ASM cell proliferation. Leptin promoted VEGF release by human ASM cells, while adiponectin did not influence VEGF release. Neither leptin nor adiponectin influenced MCP-1 secretion from human ASM cells. Adiponectin and MIP-1alpha were not secreted by human ASM cells. CONCLUSION: Human ASM cells expressed adiponectin receptors. However, adiponectin did not regulate human ASM cell proliferation or VEGF release, while leptin stimulated VEGF release by human ASM cells.

Effect of platelet-activating factor on cell proliferation & NF-kappaB activation in airway smooth muscle cells in rats.

BACKGROUND & OBJECTIVE: Despite the established pro-inflammatory actions of platelet activating factor (PAF) observed on chronic obstructive pulmonary disease (COPD), its action on airway remodeling has been still unclear. It has been reported that nuclear factor-kappa B (NF-kappaB) activity is necessary for ASMC proliferation. Further, PAF has been identified as the proximal inducer of NF-kappaB. The present study was thus aimed to investigate the effect of PAF on airway smooth muscle cells (ASMC) proliferation and to evaluate the potential role of NF-kappaB in this regulation. METHODS: Healthy male Sprague-Dowley rats of 6-8 wk age were used for obtaining ASMCs. 3-(4,5- dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) assay was to investigate the effects of PAF on ASMC proliferation and to confirm its optimum concentration for action. Additionally, cell proliferation was also examined using cell cycle assay by flow cytometry and immunocytochemical staining for proliferating cell nuclear antigen (PCNA). And NF-kappaB DNA-binding activity was assayed by electrophoretic mobility shift assay (EMSA). RESULTS: PAF stimulated ASMC proliferation with its peak at 100 nM. At this optimum concentration, PAF significantly increased the cell proliferation index (PI) and the PCNA-positive rate in the ASMCs, as well as NF-kappaB DNA- binding activity. Whereas, 20 mM N-acetylcysteine (NAC) pre-treatment effectively blocked all of these events. INTERPRETATION & CONCLUSION: The present findings demonstrated that PAF could promote ASMC proliferation, suggesting its potential involvement in airway remodeling. Our study also suggested the promising action of 20 mM NAC on the alleviation of airway remodeling due to direct inhibition of ASMC proliferation. The involved mechanism would be relevant to the change of NF-kappaB activation in ASMCs.

Achados endoscópico e citológico das vias respiratórias de potros puro sangue inglês em início de treinamento no Jóquei Clube do Paraná / Respiratory endoscopic and cytological findings in english thoroughbred foals in the beginning of their training at Jóquei Clube do Paraná / Hallazgos endoscópicos y citológicos de las vias respiratorias de potros purasangre inglés a principio de entrenamiento en el jóquei clube do paraná

O sistema respiratório de dezesseis potros PSI, com dois anos, em início de treinamento no Jóquei Clube do Paraná, foi avaliado através da videoendoscopia e citologia do aspirado traqueobrônquico. Observou-se que em sete destes animais (43,75%) foi diagnosticada hiperplasia folicular linfóide de grau III (intenso) e, nos demais (56,25%), grau II (moderado). No momento do exame, houve deslocamento do palato mole de forma persistente em dois animais (12,5%) e de forma intermitente, em outros três (18,75%). Foi constatada epiglote com aspecto frágil em apenas um animal e observada hemiplegia laríngea (grau II) em outros dois casos. Ainda, constatou-se secreção traqueobrônquica em dez animais (62,5%), em sete dos quais (70%) foi realizado aspirado e exame citológico.

The respiratory system of sixteen 2-year-old thoroughbred foals in the beginning of their training at Jóquei Club do Paraná was evaluated through videoendoscopy and tracheobronchial cytology. Seven horses (43.75%) presented (severe) level III follicular hyperplasia, and the others (56.25%) level II (moderate). At the moment of the exam there was persistent soft palate displacements in two animals (12.5%), and intermittent in three (18.75%). The epiglottis was found to be fragile in only one animal, and the level II laryngeal hemiplegia was diagnosed in two other cases. Ten animals (62.5%) presented tracheobronchial secretion, and the aspirate and cytological exams were carried out in seven (70%).

Disquinesia ciliar primaria: experiencia en 6 pacientes / Primary ciliary dyskinesia: a experience in 6 patients

Background: Primary ciliary dyskinesia is characterized by a congenital alteration of the ciliary ultrastructure and function. As a consequence, their respiratory tract sweeping action is lost and recurrent respiratory infections ensue. Aim: To analyze a clinical series of patients with primary ciliary dyskinesia, their clinical and laboratory features. Patients and methods: a retrospective review of patients with primary ciliary dyskinesia seen a university hospital, between 1994 and 1998. Bronchial biopsies were obtained with 3.6 mm diameter Olympus fibrobronchoscope, using a cayman type forceps. Ultrastructural alterations of respiratory tract ciliated cells were recorded. Results: six patients (four male) aged 9 months to 13 years old were reviewed. Three patients had situs inversus. All had repeated bouts of obstructive bronchitis and pneumonia, five had sinusitis, four atelectasis, three recurrent otitis and three had bronchiectasis. Cystic fibrosis and immunological alterations were ruled out in five children. Ultrastructural analysis revealed absence of dynein arms in three cases, absence of the internal dynein arm in one, additional peripheral microtubules and absence of dynein arms in one case. Conclusions: primary ciliary dyskinesia must be considered in the differential diagnosis of recurrent respiratory infections. Ultrastructural analysis of ciliary structure can be done in bronchial biopsies obtained through bronchoscopy

Corpos neuroepiteliais: estrutura e função no aparelho respiratório / Ultrastructure and function of neuroepithelial bodies in the respiratory system

O epitélio respiratório de alguns vertebrados, como o homem, revela a presença de células neuroendócrinas que parecem funcionar como quimioreceptores sensíveis à hipoxia, denominados corpos neuroepiteliais (CNE). Esta revisão visa abordar alguns aspectos fisiológicos e patológicos dos CNE. Discutimos sua ultra-estrutura, relação com tecidos adjacentes, inervação, hormônios relacionados e a função destas células no sistema respiratório. Tentaremos elucidar mais sobre um tema pouco pesquisado e que pode ter grande importância clínica no futuro

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