Comparative Toxicity of Nanomaterials to Air-blood Barrier Permeability Using an In Vitro Model / 生物医学与环境科学（英文）

OBJECTIVE@#To comparatively study the toxicity of four metal-containing nanoparticles (MNPs) and their chemical counterparts to the air-blood barrier (ABB) permeability using an in vitro model.@*METHODS@#ABB model, which was developed via the co-culturing of A549 and pulmonary capillary endothelium, was exposed to spherical CuO-NPs (divided into CuO-40, CuO-80, and CuO-100 based on particle size), nano-Al2O3 (sheet and short-rod-shaped), nano-ZnO, nano-PbS, CuSO4, Al2(SO4)3, Zn(CH3COO)2, and Pb(NO3)2 for 60 min. Every 10 min following exposure, the cumulative cleared volume (ΔTCL) of Lucifer yellow by the model was calculated. A clearance curve was established using linear regression analysis of ΔTCL versus time. Permeability coefficient (P) was calculated based on the slope of the curve to represent the degree of change in the ABB permeability.@*RESULTS@#The results found the increased P values of CuO-40, CuO-80, sheet, and short-rod-shaped nano-Al2O3, Al2(SO4)3, and Pb(NO3)2. Among them, small CuO-40 and CuO-80 were stronger than CuO-100 and CuSO4; no difference was observed between Al2(SO4)3 and sheet and short-rod-shaped nano-Al2O3; and nano-PbS was slightly weaker than Pb(NO3)2. So clearly the MNPs possess diverse toxicity.@*CONCLUSION@#ABB permeability abnormality means pulmonary toxicity potential. More studies are warranted to understand MNPs toxicity and ultimately control the health hazards.

Avaliação do citoesqueleto e da barreira endotelial pulmonar na malária experimental / Evaluation of the cytoskeleton and pulmonary endothelial barrier in experimental malaria

Infecções por Plasmodium sp. podem levar a um quadro respiratório grave, com complicações pulmonares denominadas lesão pulmonar aguda e síndrome do desconforto respiratório agudo (LPA/SDRA). Inflamação aguda, lesão do endotélio alveolar e do parênquima pulmonar, disfunção e aumento da permeabilidade da barreira alvéolo-capilar e, consequente, formação de edema, caracterizam esta síndrome. O modelo experimental, que utiliza o parasita murino Plasmodium berghei ANKA e camundongos da linhagem DBA/2, é empregado no estudo de mediadores imunológicos e fatores que propiciam o estabelecimento das lesões pulmonares associados à LPA/SDRA. Diversos estímulos podem atuar diretamente no aumento da permeabilidade endotelial por meio da desestabilização dos microtúbulos, rearranjo dos microfilamentos de actina e contração das células endoteliais, via sinalização de Rho-GTPases, causando disfunção da barreira endotelial. Desta forma, este trabalho tem como objetivo avaliar as alterações do citoesqueleto em células endoteliais primárias pulmonares de camundongos DBA/2 (CEPP-DBA/2), as vias de sinalização das principais Rho-GTPases e o estresse oxidativo, causados pela presença de eritrócitos parasitados com esquizontes de P. berghei ANKA (EP-PbA). As CEPP-DBA/2 foram estimuladas com TNF, VEGF ou IFNγ, em diferentes tempos de exposição, seguido da incubação com EP-PbA. Assim, foram realizados ensaios de imunofluorescência para análise do rearranjo de microfilamentos de actina e da desestabilização de microtúbulos. As vias de sinalização das Rho-GTPases foram avaliadas por Western blot, para as expressões proteicas de RhoA, Cdc42 e MLC. Além disso, ensaio fluorométrico foi realizado para detectar a produção de espécies reativas de oxigênio, resultantes do estímulo com eritrócitos parasitados. CEPP-DBA/2 estimuladas por EP-PbA, VEGF, TNF ou IFNγ, em associação ou não, apresentaram alterações morfológicas nos microfilamentos de actina e aumento dos espaços interendoteliais. Imagens de imunofluorescência também mostram desestabilização de microtúbulos e desfosforilação de FAK, causadas por EP-PbA. Os ensaios de permeabilidade validam que os eritrócitos parasitados com formas maduras de P. berghei induziram aumento da permeabilidade microvascular nas CEPP-DBA/2. Além disso, estas células, estimuladas com EP-PbA, demonstraram elevada produção de espécies reativas de oxigênio (EROs), o que pode estar contribuindo com o desenvolvimento de estresse oxidativo e com a injúria endotelial, assim como, com o aumento da permeabilidade vascular. O mais interessante é que estas alterações endoteliais podem estar relacionadas ao aumento da razão RhoA/Cdc42, da expressão proteica de MLC fosforilada e do sinal de ativação de RhoA. Em conjunto, estes resultados mostram envolvimento dos eritrócitos parasitados com esquizontes de Plasmodium berghei ANKA na desorganização do citoesqueleto e na disfunção da barreira alvéolo-capilar, via RhoA/Rho-kinase, o que pode estar contribuindo com a patogênese da LPA/SDRA associada à malária

Infections by Plasmodium sp. can lead to a serious respiratory condition with pulmonary complications, named acute lung injury and acute respiratory distress syndrome (ALI/ARDS). Acute inflammation, alveolar endothelium and lung parenchyma injuries, dysfunction and increased permeability of the pulmonary alveolar-capillary barrier and consequent formation of edema characterize this syndrome. Several stimuli can directly increase endothelial permeability through actin microfilaments rearrangement, via Rho- GTPases signaling, leading to endothelial barrier dysfunction. DBA/2 mice infected with Plasmodium berghei ANKA develop ALI/ARDS similar to that observed in humans. The purpose of this research was to assess cytoskeletal changes in DBA/2 mice primary microvascular lung endothelial cells (PMLEC), verify the signaling pathways of the Rho- GTPases and analyze the oxidative stress on these cells in the presence of P. berghei ANKA-infected red blood cells (PbA-iRBC). PMLEC were stimulated by TNF, VEGF or IFNγ followed by incubation with PbA-iRBC. Immunofluorescence assays were performed to analyze actin microfilaments rearrangement and microtubules destabilization. Western blot for RhoA, Cdc42 and MLC proteins were conducted to assess alterations in signaling pathways of Rho-GTPases. In addition, a fluorimetric assay was performed to detect the production of reactive oxygen species resulting from PbA-iRBC stimulus. P. berghei ANKA, VEGF, TNF and IFNγ stimuli, in association or not, caused morphological disturbances in actin microfilaments of PMLEC and an increase of intercellular spaces. Moreover, immunofluorescence images showed microtubules destabilization and FAK dephosphorylation in these cells, caused by PbA-iRBC. The permeability assay showed that PbA-iRBC induced an increase of microvascular permeability in PMLEC. In addition, PMLEC stimulated by PbA-iRBC, showed elevated production of ROS, which may be contributing to oxidative stress and increasing the damage of endothelial cells, as well as an increase of vascular permeability. Interestingly, these endothelial changes may be related to the increased RhoA/Cdc42 protein expressions ratio, augmented protein expression of phosphorylated MLC and RhoA activation signal. Taken together, these data demonstrate the involvement of P. berghei ANKA-infected red blood cells in cytoskeleton disorganization and alveolar-capillary barrier dysfunction, through of RhoA / Rho-kinase signaling pathway, which may contribute to ALI/ARDS pathogenesis

High postoperative serum levels of surfactant type B as novel prognostic markers for congenital heart surgery / Níveis séricos pós-operatórios tipo surfactante B elevados como novos marcadores prognósticos para cirurgia cardíaca congênita

Objective: Congenital heart diseases are observed in 5 to 8 of every 1000 live births. The presence of a valuable biomarker during the surgical periods may aid the clinician in a more accurate prognosis during treatment. Methods: For this reason, surfactant protein B plasma levels may help to evaluate patients with cardiac problems diminishing the alveolocapillary membrane stability. In this study, plasma levels of this biomarker were measured in the preoperative and postoperative periods. This study was conducted to detect the differences between pulmonary hypertensive and normotensive patients. The differences before and after cardiopulmonary bypass were examined. Results: The differences in cardiopulmonary bypass time, cross-clamp time , inotropic support dose, and duration of intensive care of patients with and without pulmonary hypertensive were found to be statistically significant (P<0.05). The results revealed that this pathophysiological state was related to other variables that were studied. We believe that the differences in preoperative and postoperative SPB levels could be attributed to alveolocapillary membrane damage and alveolar surfactant dysfunction. We found that this pathophysiological condition was significantly associated with postoperative parameters. Conclusion: The findings of the current study showed that surfactant protein B was present in the blood of patients with a congenital heart disease during the preoperative period. Long by-pass times may exert damage to the alveolocapillary membrane in patients with pulmonary hypertension and preoperative heart failure, and it is recommended to keep the option of surfactant therapy in mind during the postoperative course at the intensive care unit before preparing the patients for extubation. .

Objetivo: As cardiopatias congênitas são observadas em 5 a 8 em cada 1.000 nascidos vivos. A presença de um biomarcador importante durante os períodos cirúrgicos pode auxiliar o clínico a um prognóstico mais preciso durante o tratamento. Métodos: Por esta razão, os níveis plasmáticos de proteína B do surfactante podem ajudar a avaliar os pacientes com problemas cardíacos, diminuindo a estabilidade da membrana alvéolo-capilar. Neste estudo, os níveis plasmáticos deste biomarcador foram medidos nos períodos pré-operatório e pós-operatório. Este estudo foi realizado para detectar as diferenças entre pacientes hipertensos e normotensos em nível pulmonar. As diferenças antes e depois da circulação extracorpórea foram examinadas. Resultados: As diferenças no tempo de circulação extracorpórea, tempo de pinçamento, a dose de drogas vasoativas, e a duração da terapia intensiva de pacientes com e sem hipertensão pulmonar foram estatisticamente significativas (P<0,05). Os resultados revelaram que este estado fisiopatológico foi relacionado a outras variáveis que foram estudadas. Acreditamos que as diferenças nos níveis de SPB pré-operatório e pós-operatório pode ser atribuída a danos na membrana alvéolo-capilar e disfunção do surfactante alveolar. Descobrimos que esta condição fisiopatológica foi significativamente associada com parâmetros pós-operatórios. Conclusão: Os resultados do estudo mostraram que a proteína B surfactante estava presente no sangue de pacientes com doença cardíaca congênita no pré-operatório. Longos tempos de circulação extracorpórea podem exercer danos na membrana alvéolo-capilar em pacientes com ...

Efeito do tempo de aplicação da pressão positiva expiratória sobre a depuração pulmonar do 99mTc-DTPA / Effect of application time of positive expiratory pressure on lung clearance of 99mTc-DTPA

Introdução: A taxa de depuração pulmonar do 99mTcDTPAconstitui um índice da permeabilidade pulmonar. A aplicação dapressão positiva expiratória (EPAP), um recurso fisioterapêutico,promove aumento da expansão pulmonar, entretanto são escassosos estudos abordando a associação entre o tempo de aplicação desterecurso e a permeabilidade pulmonar. Objetivo: Comparar o tempode depuração pulmonar do 99mTc-DTPA no 15º e 30º minuto deaplicação da EPAP nas pressões de 10 e 15 cmH2O. Hipotetiza-seque a aplicação de apenas 15 minutos da técnica já poderia influenciarpositivamente o aumento do volume pulmonar. Material e métodos:Mensurou-se a taxa de depuração pulmonar do 99mTc-DTPAna posição sentada em indivíduos hígidos (n = 20) respirando sob oefeito de 10 e 15 cmH2O de EPAP, após avaliou-se o comportamentodo T1/2 no 15º e 30º minuto. Resultados: Observou-se o aumentoda taxa de depuração quando 15 cmH2O de EPAP foi aplicada (p= 0,012), porém não houve alteração com 10 cmH2O (p = 0,064).Não houve diferença estatisticamente significativa no T1/2 do 99mTc--DTPA do 15º minuto (p = 0,182) para o 30º (p = 0,489). Conclusão:Os resultados demonstraram o efeito de 15 cmH2O da EPAP noaumento da depuração pulmonar do 99mTc-DTPA sugerindo nãohaver diferença entre o 15º e 30º minuto.

Introduction: The pulmonary clearance rate of 99mTc-DTPA isindex of lung permeability. The application of expiratory positiveairway pressure (EPAP), a physical therapy tool, promotes increasein pulmonary volume, however there are few studies addressingthe association between the application time of this resource andlung permeability. Objective: To compare the time of 99mTc-DTPAlung clearance in the 15º and 30º minute of EPAP at pressures 10cmH2O and 15 cmH2O. It is hypothesized that the applicationof 15 minutes of EPAP could positively influence the increase inpulmonary volume. Methods: We measured the pulmonary clearancerate of 99mTc-DTPA in a sitting position in healthy individuals (n= 20) breathing under the effect of 10 and 15 cmH2O of EPAP,after that we evaluate the behavior of T1/2 at 15º and 30º minute.Results: We observed an increase in clearance rate with 15 cmH2OEPAP (p = 0,012), but there was no change with 10 cmH2O (p= 0,064). There was no statistically significant difference in T1/2 of99mTc-DTPA in the 15º minute (p = 0,182) to the 30º minute (p =0,489). Conclusion: The results demonstrated the effect of 15 cmH2OEPAP in the increase of the pulmonary clearance of 99mTc-DTPAsuggesting no difference between 15º and 30º minutes.

Entendendo a estrutura microscópica e submicroscópica da barreira alvéolo-capilar na lesão pulmonar aguda / Understanding the microscopic and submicroscopic structure of the alveolar-capillary membrane in acute lung injury

A lesão pulmonar aguda/síndrome do desconforto respiratório agudo (LPA/SDRA) pode ser induzida por diferentes causas. A lesão pulmonar ocorre por efeito direto sobre as células epiteliais pulmonares ou em decorrência de efeito indireto sobre as células endoteliais, onde o dano pulmonar decorre da liberação de mediadores inflamatórios em órgãos distais. Neste artigo, enfatizamos as diferenças microscópicas e submicroscópicas no pulmão envolvido na LPA e SDRA, ambas caracterizadas por intensa resposta inflamatória local com acúmulo de diferentes tipos celulares.O remodelamento do parênquima pulmonar caracterizado por fibroelastogênese ocorre em paralelo com o processo inflamatório. O prognóstico do paciente dependerá da resolução do evento inicial e do balanço entre a intensidade das respostas inflamatória e de remodelamento. Diferentes protocolos tentam modificar ambas as respostas, mas todos com resultados negativos. Postulamos que, para uma melhor compreensão da fisiopatologia da SDRA, diferenças microscópicas e submicroscópicas devem ser consideradas. Logo, para se estabelecer uma conduta clínica mais precisa e melhorar o prognóstico desses pacientes, deve-se considerar a etiologia da LPA/SDRA.

Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) can be induced by various causes. Lung injury can occur through direct or indirect effects on the lung epithelial cells, the latter caused by the release of inflammatory mediators in distal organs. In this article, we emphasize the lung microscopy and ultrastructural changes seen in ALI and ARDS, both of which are characterized by an intense inflammatory process with cell infiltration. The lung parenchyma remodeling process is characterized by fibroelastogenesis occurring in parallel with the inflammatory process. The prognosis depends on the resolution of the initial event and on the balance between the inflammatory response and the remodeling process. Although various protocols have been developed in attempts to modify those aspects, none have produced positive results.We postulate that a better understanding of ARDS cannot be gained without taking lung microscopy and ultrastructural analysis of the lung parenchyma into account. Therefore, in order to improve the clinical management and the prognosis of patients with ALI/ARDS, the etiology of the syndrome should be considered.

Como determinar a volemia em pacientes com lesão pulmonar aguda/síndrome do desconforto respiratório agudo / Determining blood volume in patients with acute lung injury/acute respiratory distress syndrome

A morbidade e a mortalidade de pacientes com lesão pulmonar aguda (LPA) ou síndrome do desconforto respiratório agudo (SDRA) permanecem elevadas. Devido às alterações na membrana alvéolo-capilar, assim como uma possível elevação da pressão hidrostática, a reposição volêmica passa a ter extrema importância. Para tal, são necessárias avaliações precisas do estado volêmico e da predição da resposta hemodinâmica.Estudos recentes enfatizam a indução de um balanço negativo em pacientes com LPA que não se encontram em choque circulatório. Novos estudos têm focado em alternativas menos invasivas e mais precisas na abordagem de fluidos em pacientes com LPA/SDRA. Nesse contexto, recentes estudos têm demonstrado a superioridade de parâmetros dinâmicos sobre parâmetros estáticos na responsividade de fluidos. Além disso, parâmetros que forneçam informações importantes de pré-carga cardíaca e do grau de edema pulmonar têm sido enfatizados por alguns ensaios clínicos de pequeno porte.Indica-se a restrição de fluidos em pacientes com LPA/SDRA desde que não haja choque circulatório. Nessa situação, considera-se o uso de parâmetros dinâmicos para determinar a quantidade e o tipo de fluido administrados. Para a determinação de edema pulmonar, lança-se mão da medida da água extravascular pulmonar, sabendo de suas potencialidades e limitações.

The morbidity and mortality remain elevated in patients with acute lung injury/acute respiratory distress syndrome (ALI/ARDS). Hemodynamic stabilization in such patients may require fluid resuscitation, but the increase in hydrostatic pressure may worsen lung edema in presence of increased permeability of the alveolar-capillary membrane. Therefore, accurate evaluation of the fluid state and prediction of the hemodynamic response are essential. Recent studies have focused on the induction of a negative fluid balance in ALI patients who are not in circulatory shock. Other studies of ALI/ARDS patients have focused on fluid management strategies that are less invasive and more accurate. In this context, recent studies have demonstrated the superiority of dynamic parameters over static parameters in determining the hemodynamic response. In addition, parameters that furnish useful data regarding cardiac preload and the degree of pulmonary edema have been emphasized in recent small clinical trials. In ALI/ARDS patients, fluid restriction is indicated if there are no clinical signs of circulatory shock. In such cases, the nature and quantity of fluid administered should be determined on the basis of the dynamic parameters. To screen for pulmonary edema, extravascular lung water can be measured, assuming that its potential and limitations are borne in mind.

Diffusing capacity for lung carbon monoxide [DLCO] in chemical lung injuries due to the use of mustard gas in the poisoned soldiers of Iran-Iraq war 2006

To assess the Mustard gas exposure effects on pulmonary system, particularly on diffusing capacity for lung carbon monoxide [DLCO] and simple spirometry. Sixty-five sulfur mustard- poisoned soldiers from Mostazafan and Janbazan organization were referred to our center in 2005. Complete history, physical examination, chest X ray, Echocardiography, Arterial blood gas, high - resolution computerized tomography, diffusion capacity for lung carbon monoxide and spirometry of these were performed and compared this result with normal value. The mean value of indices in studied injured subjects was: Spirometry: forced expiratory volume in one second [FEV1] = 70.4, Forced vital capacity [FVC] = 66.5, EFE 25-75=81.1, FEV1/FVC=101.9, Flow 25% = 28.7, Flow 50%= 72.9, Flow 75%= 100.1, Sample volume: Functional residual capacity of lungs [FRC] = 131.5, residual volume [RV] = 157.3, RV/TLC= 169.1, Total lung capacity [TLC] = 91.3, KCO= 131.6, TLCO= 116.3. No significant correlation was observed between TLCO values with HRCT, echocardiography, ABG and spirometry values [P>0.05]. We recommend TLCO and RV/TLV tests to assess severity of Injuries as there is no a suitable criterion to measure the real consequences of mustard gas on affected combatants and Biological markers are also needed to determine cause- effect relations

Morphological Study of Respiratory Portion of Mouse Lung following Chemical Glue-Vapor Inhalation / 대한해부학회지

Industrial glues contain many kinds of organic solvents and glue sniffing by young people has become a social problem in Korea. Glue vapor may induce chronic toxicities different from those induced by exposures to the solvent of single component. We studied the effects of the inhalation of glue vapor on the primary target organ, the pulmonary epithelium of the respiratory system. Vapor samples of glue were collected for analysis; the components were acetone, n-hexane, methyl cyclopentane, c-hexane, and toluene. For the inhalation of glue vapor, experimental mice were exposed in a whole body chamber for 20 min/d for 3, 5, 7, and 14 d. Control groups were exposed to room air. Animals were euthanized and lung tissues were fixed in 10% neutral formalin for light microscopy, and in 2.5% glutaraldehyde plus 1.5% paraformaldehyde for electron microscopy. The results are as follows. 1. Alcianophilic bands were not detected in the normal alveolar epithelium, but weak alcianophilic bands were detected in bronchioles. Alcian blue-PAS and PAS positive cells were found in the mucosae of mice exposed to glue vapor for 5 and 7 d. 2. Types I and II pneumocytes and capillary endothelial cells were found in the normal alveolar epithelium. The blood-air barrier consists of Type I pneumocytes, a common basal lamina, and the capillary endothelium. 3. The alveolar epithelium of vapor-exposed mice showed more type II pneumocytes. In the longerexposed group, Type I pneumocytes and endothelial cells contained many pinocytotic vesicles. 4. The vapor-exposed lungs showed macrophages in the alveolar space, mild interstitial swelling, and increased numbers of collagenous fibers. Clearly, ultrastructural changes in pulmonary epithelia can occur following glue sniffing.

Ultrastructure of the camel's pulmonary alveoli with special reference to the air-blood barrier [with 7 figures]

The present study was conducted to characterize the cellular population lining the alveoli of the camel's lung. It focused on the ultrastructural findings and their reflection on the physiological role in gas exchange and transcytosis through the air-blood barrier [ABB]. It was carried on adult camel using electron microscope. Ultrastructural examination revealed that the pulmonary alveoli were lined with a continuous epithelium comprising two major cell types; the predominant, attenuated pneumocyte type I and the less popular, irregularly cuboidal pneumocyte type II. Two 'forms of fibroblasts were distinguished; the most remarkable feature of the first form was its well-developed and abundant rough endoplasmic reticulum. The second form was characterized by large, irregular, dark stained nucleus and little amount of cytoplasm. The most obvious feature of endothelial cells was the concentration of small vesicles [pinocytotic vesicles] adjacent to the endothelial cell membranes. They were circumscribed by a continuous basal lamina. Along the same endothelial cell, two cytoplasmic zones were existing; a thin cytoplasmic area containing few or no plasmalemmal vesicles [avesicular area] and another thicker cytoplasmic area with numerous plasmalemmal vesicles and endocytotic pits [vesicular area]

Protective effects of bactericidal/permeability increasing protein simulated peptide on murine acute lung injury induced by lipopolysaccharide / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the protective effects of bactericidal/permeability increasing protein simulated peptide (bactericidal neutralizing endotoxin protein, BNEP) on murine acute lung injury (ALI) induced by lipopolysaccharide (LPS).</p><p><b>METHODS</b>A murine model of ALI was reproduced by lipopolysaccharide via intranasal instillation. The Balb/c mice were randomly divided into control (n = 20, with nasal instillation of isotonic saline), LPS instillation (n = 20, with nasal instillation of isotonic saline and LPS) and BNEP treatment (n = 20, with nasal instillation of isotonic saline plus LPS and BNEP) groups. The ratio of lung wet weight to dry weight, the permeability of pulmonary capillary vessels and the histopathology of pulmonary tissue were determined in all groups. The change in the expression of Toll-like receptor 2 and 4 (TLR2/4) in the pulmonary tissue was detected by immunohistochemistry.</p><p><b>RESULTS</b>Compared with LPS instillation group, the ratio of lung wet weight to dry weight and the permeability of pulmonary capillary vessel was decreased significantly in the BNEP group, and the inflammatory infiltration in the pulmonary tissue induced by neutrophil influx was alleviated markedly with BNEP treatment. The expression of TLR2 and TLR4 in pulmonary vascular endothelial cells, macrophages and alveolar type II epithelial cells in BNEP group were lower than those in LPS group (TLR2: 128 +/- 10 vs 214 +/- 12, P < 0.01).</p><p><b>CONCLUSION</b>BNEP, as a simulated peptide of BPI, exerted a remarkable protective effect on ALI induced by LPS.</p>

Expressions of Laminin-1 in Lung Alveolar Septa after CS gas Exposure in Rats / 결핵및호흡기질환

BACKGROUND: Laminin-1 is known to have regular functions in the development and course of differentiation of the lungs. The morphogenesis and distribution of laminin-1 still remains as a mystery and its distribution and changes in the molecular structure of laminin-1 in the pathogenesis of the lung still is a subject of great controversy. In this study, experiments were done to delineate the distribution and changes in the amount of laminin-1 after inducing inflammation of the lungs by exposing experimental animals to CS gas and especially, to find compositions of laminin-1 within type II pneumocytes. MATERIALS AND METHODS: The experimental subjects of study were newborn rats and the extracted tissue from the experimental rats were viewed under light microscope and electron microscope after the sections were treated with immunohistochemical methods and immunogold reaction methods using bounded gold particles. RESULTS: 1) Lymphocytes and mononuclear phagocytes invaded the alveolar septa in the 2 day group rats after CS gas exposure and intense interstitial inflammation was seen in the 3 day group. 2) Laminin immunoreactions decreased to a moderate degree in the 2 and 3 day group rats after CS gas exposure and strong laminin immunoreactions were seen again in the 5 and 7 day group rats. 3) Gold particles in basal lamina of the lung blood-air barrier decreased and in the type I pneumocytes decreased in the 2 and 3 day group rats after CS gas exposure. 4) Gold particles were seen only on the surface of the cell membranes of type II pneumocytes of the 1 and 2 day group rats after CS gas exposure. 5) Few gold particles around the lamellar bodies and cytoplasm of type II pneumocytes in the control rat group and at 12 hours after CS gas exposure. Gold particles are seen only on the surface of type II pneumocytes of the 1 and 2 day group rats after CS gas exposure and are evenly distributed in small amounts in the cells of the 3 day group after CS gas exposure. CONCLUSION: CS gas exposure in the rats caused inflammation of lung alveolar septa and also induced a decrease in laminin-1 in basal lamina and loss of laminin-1 in the cytoplasm of type II pneumonocytes. As the inflammatory cells disappeared, an increase in the distribution of laminin-1 occurred. This reflects tissue regeneration functions of laminin-1 in the pneumocytes of rats and the distribution of laminin-1 in type II pneumocytes can be seen through the electron microscope using immunogold methods.

Effects of sevoflurane on membrane permeability of alveolar capillaries in rats with acute lung injury caused by endotoxin / 中华外科杂志

<p><b>OBJECTIVE</b>To observe the effect of sevoflurane on membrane permeability of alveolar capillaries in rats with acute lung injury and the ratio of inflammatory cells in bronchoalveolar lavage fluid in rats with acute endotoxin lung injury.</p><p><b>METHODS</b>48 Wistar rats were selected and divided into group C, L, S1L and S2L after injection evans blue 50 mg/kg in random with 12 rats in each group. Group C was taken as control group, 1.2 ml normal saline was injected into the rats via femoral vein and then the rats were mechanically ventilated for 4 hours; The rats in group L were also mechanically ventilated for 4 hours after injection of endotoxin 5 mg/kg via the same vein. For the rats in group S1L and S2L, 1.0 or 1.5 minimal alveolar concentration (MAC) sevoflurane was inhaled with mechanical ventilation after injection of endotoxin 5 mg/kg. Evans blue was not injected into 6 rats of each group in order that the 6 rats could be used for pathological examination and alveoli lavage, lung pathomorphological score of the lung, lung wet/dry weight ratio, the content of lung water, lung permeability index, content of evans blue, total amount and ratio of inflammatory cells in bronchoalveolar lavage fluid (BALF) were all determined.</p><p><b>RESULTS</b>Sevoflurane of 1.0 MAC and 1.5 MAC made lung wet/dry weight ratio and content of lung water change insignificantly; lung permeability index, content of evans blue and pathomorphological score in group S1L decreased from 4.86 +/- 0.82, 112.21 +/- 11.44 ng/mg, 9.17 +/- 0.90 to 3.98 +/- 0.50, 92.85 +/- 11.80 ng/mg, 7.50 +/- 0.96; group S2L decreased to 3.91 +/- 0.34, 96.33 +/- 8.79 ng/mg, 7.67 +/- 0.75. Sevoflurane of 1.0 MAC and 1.5 MAC did not have a significantly effect on total amount and ratio of inflammatory cells in BALF.</p><p><b>CONCLUSION</b>Membrane permeability of alveolar capillaries after acute endotoxin lung injury decreased by inhalation of sevoflurane of 1.0 MAC and 1.5 MAC and pathological injury of lung tissue relieved.</p>

Inhibition of Viability and Genetic Change in Hypoxia-treated Lung Pericytes / 결핵및호흡기질환

BACKGROUND: Lung pericytes are important constituent cells of blood-air barrier in pulmonary microvasculature. These cells take part in the control of vascular contractility and permeability. In this study, it was hypothesized that change of lung pericytes might be attributable to pathologic change in microvasculature in acute lung injury. The purpose of this study was how hypoxia change proliferation and genetic expression in lung pericytes. METHODS: From the lungs of several Sprague-Dawley rats, performed the primary culture of lung pericytes and subculture. Characteristics of lung pericytes were confirmed with stellate shape in light microscopy and immunocytochemistry. 2% concentration of oxygen and 200muM CoCl2 were treated to cells. Tryphan blue method and reverse transcription-polymerase chain reaction were done. RESULTS: 1. We established methodology for primary culture of lung pericytes. 2. Hypoxia inhibited cellular proliferation in pericytes. 3. Hypoxia could markedly induce vascular endothelial growth factor(VEGF) and smad-2. 4. Hypoxia-inducible factor-1alpha (HIF-1alpha)was also induced by 2% oxygen. CONCLUSION: Viability of lung pericytes are inhibited by hypoxia. Hypoxia can stimulate expression of hypoxia-responsive genes. Pericytic change may be contributed to dysfunction of alveolar-capillary barrier in various pulmonary disorders.

Utilidad de la medición de permeabilidad alveolocapilar con TC99m-DTPA en el compromiso pulmonar de pacientes portadores del virus de inmunodeficiencia adquirida / Value of Tc99m-DTPA alveolar permeability in lung involvement detection of patients with HIV infection

Se evaluó la utilidad de la medición de permeabilidad alveolocapilar con Tc99m -DTPA en pacientes VIH positivo con posible compromiso pulmonar e infección por pneumocystis carinii (PC) . Se incluyeron 20 pacientes con síntomas respiratorios y 4 con síntomas sistémicos usando como control a un grupo de 11 asintomáticos con similar valor de linfocitos CD, todos con suspensión de tabaco previa. Se realizaron radiografía de tórax, hemograma, esputo inducido y/o fibrobroncoscopía, obteniéndose confirmación de presencia o ausencia de PC en 16 pacientes sintomáticos y 3 asintomáticos. Para detección de PC la sensibilidad fue 78 por ciento, la especificidad 40 por ciento y la seguridad diagnóstica 58 por ciento. Para procesos inflamatorios pulmonares los valores fueron 85 por ciento, 60 por ciento y 79 por ciento, respectivamente. Cuatro de seis pacientes falsos positivos para PC tenían cuadros que explicaban la alteración del DTPA. Concluyendo, el DTPA es sensible pero poco específico para detectar infección pulmonar por PC, siendo superior para procesos inflamatorios

The Ultrastructural Development of Air-Blood Barrier in Human Fetal Lung / 대한해부학회지

The air-blood barrier represents the maturity of developing lung. The development of air-blood barrier in human fetal lung was studied by transmission electron microscopy. The results obtained were as follows. 1. The formation of air-blood barrier started at 16 week of postcoitum, which was the end of pseudoglandular period. The basement membranes began to be fused with each other as the capillaries penetrated between epithelial cells of primitive alveoli. 2. The flattening of the type II alveolar cells was observed only around the site of fused basement membranes, which seemed to be developed not by mechanical force but by induction of the fused basement membrane. 3. The basement membranes of capillaries and alveoli were relatively flat until the fusion occurred, but they showed severe folds with the occurrence of fusion. But with the proceeding of the terminal sac period, the folds greatly decreased. In summary, the air-blood barrier began to develop at the end of pseudoglandular period and was formed as capillaries penetrated the cytoplasms of epithelial cells devoided of the nuclei. The fused basement membranes seems to play an important role in the development of air-blood barrier.

Daño pulmonar con amiodarona y su seguimiento con la prueba de permeabilidad alvéolo-capilar con T c99m-DTPA: comunicación en dos casos / Pulmonary damage due to amiodarone and it follow up through alveolar-capillary permeability test: report of 2 cases

La amiodarona es una potente droga antiarrítmica de importante uso en nuestro medio, caracterizada por su prolongada vida media de eliminación. Se le ha descrito una reacción adversa pulmonar que se manifiesta como una enfermedad difusa, intersticial y/o alveolar, con una incidencia promedio de 5 a 7 por ciento de los casos. Esta alveolitis presenta una importante complejidad diagnóstica, tanto por la multiplicidad de diagnósticos diferenciales, como por la ausencia de una prueba específica y precoz. Se presentan dos casos de alveolitis por amiodarona cuyo diagnóstico y seguimiento se realizó con la prueba de Tc99m-DTPA. Esta prueba resultó significativamente disminuida (menos del 50 por ciento del valor normal) en el momento del diagnóstico y se mantuvo anormalmente baja por tiempo prolongado a pesar del tratamiento esteroidal. Se discuten algunos aspectos de esta prueba radioisotópica y sus alcances diagnósticos y de seguimiento en el daño pulmonar unducido por amiodarona

Efectos de la contaminación atmosférica sobre la permeabilidad alvéolo-capilar usando Tc99m DTPA / Effects of air pollution on alveolar-capillary permeability using 99mTc-DTPA

Es un hecho conocido que diversas condiciones pueden afectar la permeabilidad alvéolo-capilar (PAC) tales como algunas enfermedades inflamatorias activas (neumonitis, síndrome de "distress" respiratorio del adulto, membrana hialina, sarcoidosis, asbestosis, infección por neumocistis carinii y otras infecciones) y también el tabaco y el ozono. En esta revisión se analizan algunos métodos capaces de evaluar la integridad de la PAC; difusión de O2 y CO, lavado broncoalveolar, Gn 67 y tomografía axial computarizada de cortes delgados. Otra prueba no invasiva es la medición de la difusión del Tc99m-DTPA. Corresponde a una molécula pequeña, hidrosoluble (490 Dalton) la cual si es inhalada desde el micronebulizador puede depositarse en la parte distal del árbol broncoalveolar, siendo posible entonces medir su difusión al lecho capilar sanguíneo. La exposición experimental de animales a contaminantes atmosféricos altera la permeabilidad traqueal y broncoalveolar y también lo hace la exposición a O3. Santiago de Chile es una de las ciudades de mayor contaminación atmosférica, especialmente en su área central. Se comunica un trabajo preliminar acerca del efecto del smog sobre la salud de jóvenes voluntarios durante el invierno (período de mayor smog) comparando en área central de Santiago con una zona semirural no contaminada. Hubo diferencias significativas entre ambos grupos. El grupo de Santiago fue también estudiado en verano, encontrándose una diferencia estacional significativa. Con los hechos expuestos queda abierta una forma fácil de evaluar no invasivamente la población expuesta a contaminantes atmosféricos