Anti-fibrotic effects of pirfenidone and rapamycin in primary IPF fibroblasts and human alveolar epithelial cells.

BACKGROUND: Pirfenidone, a pleiotropic anti-fibrotic treatment, has been shown to slow down disease progression of idiopathic pulmonary fibrosis (IPF), a fatal and devastating lung disease. Rapamycin, an inhibitor of fibroblast proliferation could be a potential anti-fibrotic drug to improve the effects of pirfenidone. METHODS: Primary lung fibroblasts from IPF patients and human alveolar epithelial cells (A549) were treated in vitro with pirfenidone and rapamycin in the presence or absence of transforming growth factor ß1 (TGF-ß). Extracellular matrix protein and gene expression of markers involved in lung fibrosis (tenascin-c, fibronectin, collagen I [COL1A1], collagen III [COL3A1] and α-smooth muscle actin [α-SMA]) were analyzed. A cell migration assay in pirfenidone, rapamycin and TGF-ß-containing media was performed. RESULTS: Gene and protein expression of tenascin-c and fibronectin of fibrotic fibroblasts were reduced by pirfenidone or rapamycin treatment. Pirfenidone-rapamycin treatment did not revert the epithelial to mesenchymal transition pathway activated by TGF-ß. However, the drug combination significantly abrogated fibroblast to myofibroblast transition. The inhibitory effect of pirfenidone on fibroblast migration in the scratch-wound assay was potentiated by rapamycin combination. CONCLUSIONS: These findings indicate that the combination of pirfenidone and rapamycin widen the inhibition range of fibrogenic markers and prevents fibroblast migration. These results would open a new line of research for an anti-fibrotic combination therapeutic approach.

Comparacion de un nuevo agente basado en nanotecnología con el talco convencional para el control de los derrames pleurales malignos / Comparison between a new nanotechnology-based agent and conventional talc in the control of malignant pleural effusions

En este trabajo usamos dióxido de titanio (TiO2), fabricado mediante nanotecnología. Para demostrar su superioridad respecto al talco, realizamos un estudio in vitro comparando la respuesta pro-inflamatoria de ambos agentes sobre células malignas y mesoteliales benignas; investigando la posible inducción de apoptosis y la posible inhibición de angiogénesis también por ambos agentes. Realizamos cultivo de líneas celulares derivadas de mesotelio humano, procedente de mesotelioma bifásico humano y adenocarcinoma bronquial humano. Las células se co-cultivaron con diferentes dosis de talco y de nanopartículas de TiO2. En todas las muestras de sobrenadantes de los cultivos se analizaron los niveles de diferentes mediadores inflamatorios. La tasa de apoptosis se analizó por la expresión de Caspasa-3. Para el estudio de angiostasis se determinaron los niveles de endostatina mediante técnica ELISA. Observamos que la viabilidad de las células mesoteliales benignas es mucho menor al emplear TiO2. En el caso de las células mesoteliales malignas, se observó el mismo efecto con dosis alta de TiO2. En el adenocarcinoma de pulmón, la viabilidad de estas células expuestas al talco fue netamente inferior a la que se observó en la línea celular benigna. La producción de IL-8 fue mucho mayor por parte de las células mesoteliales neoplásicas que por las benignas y aumentó siguiendo un patrón dosis dependiente frente al talco, mientras que cayó con el TiO2. Según estos resultados, se demuestra que el talco es superior al TiO2 en su capacidad de producir mediadores que favorecerían la pleurodesis para el control del derrame pleural maligno

For this study, we used titanium dioxide (TiO2), produced using nanotechnology. To show its superiority with respect to talc, we completed an in vitro study comparing the pro-inflammatory response of both agents towards malignant and benign mesothelial cells; researching the possible apoptosis induction and possible inhibition of angiogenesis for both agents. We took a culture of cell lines derived from human mesothelioma, originating from human biphasic mesothelioma and human bronchial adenocarcinoma. The cells were cocultured with different doses of talc and TiO2 nanoparticles. The levels of different inflammatory mediators were analyzed for each culture supernatant sample. The apoptosis rate was analyzed using caspase-3 expression. The endostatin levels were determined for the angiostasis study using the ELISA technique. We observed that the viability of the benign mesothelial cells is much lower after using TiO2. In the case of malignant mesothelial cells, the same effect was observed with a high dose of TiO2. In adenocarcinoma of the lung, the viability of these cells exposed to talc was distinctly lower than that which was observed in the benign cell line. IL-8 production was much higher in neoplastic mesothelial cells than in benign cells and increased following a dose-dependent pattern with talc, while it decreased with TiO2. According to these results, we can see that talc is superior to TiO2 in its ability to produce mediators which favor pleurodesis for the control of malignant pleural effusions

Evaluación del efecto de la administración endobronquial de ácido tranexámico (ATx) en el sangrado de vía aérea / Assessment of the effect of endobronchial administration of tranexamic acid (ATx) in airway bleeding

INTRODUCCIÓN: la hemoptisis puede ser espontánea o iatrogénica, durante la broncofibroscopia y toma de muestras. Falta por determinar la efectividad del tratamiento antifibrinolítico con ácido tranexámico (ATx) a nivel endobronquial. OBJETIVOS: valorar la efectividad del ATx por vía endobronquial en la hemoptisis y estudiar las repercusiones del ATx sobre los parámetros de coagulación-fibrinolisis. PACIENTES Y MÉTODOS: se incluyeron 69 pacientes y se establecieron tres grupos: grupo-1 de pacientes con hemoptisis, tratados con ATx (n = 29), grupo-2 de pacientes con neoplasia de pulmón, sin tratamiento local con ATx (n = 20) y grupo-3 de pacientes sin patología neoplásica y sin tratamiento local con ATx (n = 20). Se determinaron niveles séricos basales de TAT (complejo trombina-antitrombina) y DD (dímero-D) 1 hora tras la técnica. Los grupos 2-3 no tratados con ATx, son grupos referencia para valorar la significación de datos obtenidos en los pacientes tratados con ATx. RESULTADOS: el ATx fue efectivo en el 90% de los casos. No se observaron complicaciones inmediatas. La repercusión del ATx sobre el equilibrio coagulación-fibrinólisis es evidente. Los niveles basales séricos de TAT y DD están incrementados en todos los grupos, aunque sin diferencia significativa entre estos. La diferencia en los niveles de TAT entre la determinación basal y la obtenida tras una hora, fue significativa en los tres grupos, más evidente en el grupo-1 (11x superior) tratado con ATx. CONCLUSIONES: el ATx por vía endobronquial es efectivo en la hemoptisis. Este fármaco parece influir en los parámetros de coagulación-fibrinólisis aunque de forma subclínica, dada la ausencia de complicaciones tromboembólicas clínicamente significativas

INTRODUCTION: hemoptysis could be spontaneous or iatroge nic during bronchofiberscopy and when harvesting samples. The effectiveness of anti-fibrinolytic treatment using tranexa mic acid at the endobronchial level has yet to be determined. OBJECTIVES: to assess the effectiveness of endobronchial application of ATx in hemoptysis and study the repercussions of ATx on systemic coagulation- fibrinolysis parameters. PATIENTS AND METHODS: 69 patients were included in the study and three groups were established: Group-1 included patients with hemoptysis treated with ATx (n=29), Group-2 included patients with lung neoplasia without ATx local treatment (n=20) while Group-3 included patients without neoplastic pathology and without ATx local treatment (n=20). Basal levels for TAT (thrombin-anti-thrombin complex) and DD (dimer-D) were determined and one hour after the technique was applied. Groups 2 and 3 were not treated with ATx; they were reference groups to assess the signification of data obtained in patients treated with ATx. RESULTS: ATx was effective in 90% of the cases. No immediate complications were observed. The repercussion of ATx on the coagulation-fibrinolysis is evident. Basal levels for TAT and DD in peripheral venous blood were increased in all groups, although without significant differences among them. The differences in TAT levels between the basal determination and that obtained after one hour was significant in all three groups; it was more evident in Group-1 (11 times greater) which was treated with ATx. CONCLUSIONS: endobronchial application of ATx is effective in hemoptysis. This drug could influence the coagulation-fibrinolysis parameters although sub-clinically, given the absence of clinically significant, thromboembolic complications

Interacciones entre stents metálicos desnudos y liberadores de droga con fibroblastos respiratorios co-cultivados in vitro / Interactions between bare metal and drug eluting stents co-cultivated with respiratory fibroblasts in vitro

OBJETIVO: testar una línea de fibroblastos (MRC-5) frente a distintos tipos de stents metálicos, desnudos o con liberación de droga. MÉTODOS: co-cultivamos durante tres semanas dos stents metálicos desnudos (Zilver Flex(R), aleación de níquel-titanio, niti-nol) y Wallstent(R) (aleación de cobalto), y otros tres liberadores de paclitaxel o everolimus: Zilver PTX(R) (nitinol con 3 μg/mm2 paclitaxel), Taxus Liberté(R) (acero 316-L con 1 μg/mm2 paclitaxel) y Promus(R) (aleación de cromo-cobalto con evero-limus). Se determinaron los niveles de interleuquina 8 (IL-8), factor básico de crecimiento de fibroblastos (bFGF), factor de crecimiento vascular endotelial (VEGF), y TGF-β1 (Transfor-ming growth factor-beta). RESULTADOS: se observó gran proliferación celular alrededor de los stents desnudos (Zilver Flex(R) y Wallstent(R)), aunque con distinto ritmo en el crecimiento (más tardío y más intenso en el stent de nitinol, comparado con el de cobalto), mientras que la celularidad decreció rápidamente en los stents liberadores de droga, especialmente en el Zilver PTX(R), comparado con Taxus Liberté(R). Tras un crecimiento inicial relativamente lento, la celularidad se recuperó en el Promus(R), el cual destacó por sus altos niveles de IL-8 y de TGF-β1 en fases precoces. El Taxus Liberté(R)también indujo producción marcada de IL-8 in vitro. En conclusión, ninguno de los stents que hemos testado en el presente estudio sería suficientemente seguro para prevenir su encapsulación fibrótica, aunque el stent de nitinol desnudo sería, por el momento, el menos agresivo en este sentido

OBJECTIVE: test a line of fibroblasts (MRC-5) against different types of bare metal or drug eluting stents. METHODS: we co-cultivated two bare metal stents for three weeks (Zilver Flex(R), nickel-titanium alloy, shape-memory alloy) and Wallstent(R) (cobalt alloy), and other three paclitaxel-eluting or everolimus-eluting stents: Zilver PTX(R) (shape-me-mory alloy with 3 μg/mm2 paclitaxel), Taxus Liberté(R) (steel 316-L with 1 μg/mm2 paclitaxel) and Promus(R) (chromium-cobalt alloy with everolimus). The levels of interleukin-8 (IL-8), basic fibroblast growth factor (bFGF), vascular en-dothelial growth factor (VEGF), and TGF-β1 (Transforming growth factor-beta) were determined.RESULTS: a large proliferation of cells was observed around the bare stents (Zilver Flex(R) and Wallstent(R)), although with different speeds of growth (slower and more intense with the shape-memory nitinol alloy stent, compared with that of cobalt), while the number of cells decreased rapidly with the drug eluting stents, especially the Zilver PTX(R), compared with Taxus Liberté(R). After a relatively slow initial growth, the number of cells recovered in the Promus(R), which stood out in association with its high levels of IL-8 and of TGF-β1 in early phases. The Taxus Liberté(R) also induced a marked pro-duction of IL-8 in vitro. In conclusion, none of the stents we tested in this study would be sufficiently safe to prevent their fibrotic encapsulation, although the bare shape-memory alloy stent (nitinol) would, for the time being, be the least aggressive in this regard

Expresión de aquaporinas en tejido pulmonar de pacientes con enfermedad pulmonar obstructiva crónica / Aquaporin expression in the lung tissue of copd patients

INTRODUCCIÓN: Estudio de la expresión de aquaporinas (AQP1 y AQP5) en el tejido bronquial y parénquima pulmo-nar de pacientes con enfermedad pulmonar obstructiva cróni-ca (EPOC) y fumadores sin la enfermedad. MÉTODO: Utilizando un diseño caso-control, se seleccionó un grupo de 15 pacientes con EPOC (93,3% varones, con una edad media de 68 años, una media de FEV1 del 72% y 26,7% con corticosteroides inhalados) y 15 fumadores sin la enfermedad, a los cuales se les sometió a cirugía de resección pulmonar por neoplasia pulmonar. Se estudió la expresión de AQP1 y AQP5 en el tejido bronquial y en parénquima pulmo-nar mediante reacción en cadena de la polimerasa en tiempo real.RESULTADOS: No encontramos diferencias en la expresión génica de estas AQPs en ambos territorios pulmonares entre los pacientes con EPOC y los fumadores sin la enfermedad. Sin embargo, en los pacientes EPOC, la expresión de AQP1 era 2,41 veces mayor en el parénquima comparado con los controles, mientras que la AQP5 mostraba un patrón inverso, con 7,75 veces mayor expresión en el tejido bronquial de los sujetos control.CONCLUSIÓN: Los resultados del presente trabajo proporcio-nan evidencia inicial respecto a la expresión de AQP1 y AQP5 en pacientes con EPOC

INTRODUCTION: Study of aquaporin expression (AQP1 and AQP5) in the bronchial tissue and lung parenchyma of pa-tients with chronic obstructive pulmonary disease (COPD) and smokers without the disease. METHOD: Using a case-control design, a group of 15 patients with COPD was selected (93.3% males, with an average age of 68 years, an average FEV1 of 72% and 26.7% with inha-led corticosteroids) and 15 smokers without the disease, who underwent lung resection surgery due to lung neoplasm. The expression of AQP1 and AQP5 in the bronchial tissue and in lung parenchyma was studied using real-time polymerase chain reaction (PCR). RESULTS: No differences were found in the gene expression of these AQPs in either lung territories between the patients with COPD and the smokers without the disease. Nevertheless, in the COPD patients, the expression of AQP1 was 2.41 times greater in the parenchyma compared with the controls, while the AQP5 showed an inverse pattern, with 7.75 times greater expression in the bronchial tissue of the control subject. CONCLUSION: The results of this study provide initial evidence regarding the expression of AQP1 and AQP5 in patient with COPD

Estudio de la asociación de los polimorfismos de laglutation -S-transferasa y la enfermedad pulmonar obstructiva crónica en la raza caucásica / Study of the association of polymorphisms of glutathione-s-transferase and chronic obstructive pulmonary disease in caucasians

Introducción: Diversos estudios previos han encontrado una asociación no consistente entre los polimorfismos de la Glutation-S-transferasa (GST) y la enfermedad pulmonar obstructiva crónica (EPOC), con una asociación diferente según el área geográfica estudiada a nivel mundial. El objetivo del presente trabajo fue estudiar esa relación en una muestra caucásica española. Método: Estudio observacional analítico de casos-control en el que se incluyeron pacientes con EPOC y sujetos fumadores sin la enfermedad. A cada sujeto incluido se le recogieron sus datos sociodemográficos y clínicos mediantes un cuestionario estandarizado y se les extrajo una muestra de sangre para el estudio de los polimorfismos GSTP1 Ile105Val (A131G) Exon5, GSTP1 Ala114Val exón 6, GSTM1 de elección y GGSTT1delección.Resultados: La muestra estaba compuesta por 143 casos (64años, FEV1 69%) y 55 controles. El polimorfismo más asociado fue el GSTT que en el análisis bivariante se acercó a la significación estadística, alcanzándola en el análisis de regresión ajustado por sexo, paquetes-año e IMC (p = 0,031). El análisis (..) (AU)

Introduction: Various previous studies have found a non consistent association between polymorphisms of Glutathione-S-transferase (GST) and chronic obstructive pulmonary disease (COPD), with a different association according to the geographic area studied at global level. The objective of the present work was to study that relationship in a Spanish Caucasian sample. Method: Analytical observational case-control study including patients with COPD and smokers without the disease. Socio-demographic and clinical data were recorded by means of a standardized questionnaire, and a blood sample extracted from each participant for the study of the polymorphisms (..) (AU)

Parenchymal trafficking of pleural mesothelial cells in idiopathic pulmonary fibrosis.

Idiopathic pulmonary fibrosis (IPF) is characterised by myofibroblast proliferation leading to architectural destruction. Neither the origin nor the continued proliferation of myofibroblasts is well understood. Explanted human IPF lungs were stained by immunohistochemistry for calretinin, a marker of pleural mesothelial cells (PMCs). Chronic obstructive pulmonary disease (COPD) and cystic fibrosis (CF) lungs acted as controls. The number of PMCs per 100 nucleated cells and per photomicrograph was estimated along with the Ashcroft score of fibrosis. Mouse PMCs expressing green fluorescent protein (GFP) or labelled with nanoparticles were injected into the pleural space of mice given intranasal transforming growth factor (TGF)-ß1. Mouse lungs were lavaged and examined for the presence of GFP, smooth muscle α-actin (α-SMA) and calretinin. Calretinin-positive PMCs were found throughout IPF lungs, but not in COPD or CF lungs. The number of PMCs correlated with the Ashcroft score. In mice, nanoparticle-laden PMCs were recoverable by bronchoalveolar lavage, depending on the TGF-ß1 dose. Fluorescent staining showed α-SMA expression in GFP-expressing PMCs, with co-localisation of GFP and α-SMA. PMCs can traffic through the lung and show myofibroblast phenotypic markers. PMCs are present in IPF lungs, and their number correlates with IPF severity. Since IPF presumably begins subpleurally, PMCs could play a pathogenetic role via mesothelial-mesenchymal transition.

Interleukin-8 activates coagulation and correlates with survival after talc pleurodesis.

The aim of our study was to investigate whether interleukin (IL)-8 activates systemic coagulation after talc pleurodesis in malignant pleural effusion (MPE), and whether levels of IL-8 in plasma are related to early death after talc pleurodesis. IL-8 and tumour necrosis factor (TNF)-alpha were measured in samples from 231 MPE patients before and after talc pleurodesis. Whole blood from 31 healthy volunteers was incubated with IL-8, TNF-alpha and thromboplastin for 3 h in vitro, and thrombin-antithrombin (TAT) levels were measured. The same stimulation of blood samples was repeated using doses of calibrated talc. Nine, 12 and 17 patients died within 7, 10 and 15 days respectively. IL-8 was elevated in 102 patients within 48 h, and thrombotic events were observed in six of those patients. Survival correlated inversely with IL-8 at 24 and 48 h, and a significant correlation was also found between IL-8 and TAT. A positive dose-dependent correlation with TAT production was observed when blood was stimulated with IL-8 in vitro. However, there was no significant response to stimulation with talc, as compared with control blood samples. IL-8 is involved in the activation of coagulation that may occur after talc pleurodesis, and might also be implicated in early death of patients with MPE.

Valor pronóstico de determinados marcadores biológicos en la supervivencia global y la respuesta al tratamiento del cáncer de mama con metástasis pleurales / Prognostic value of specific biological markers in the global survival rate and response to treatment of breast cancer with pleural metastasis

Objetivo: 1. Estudiar qué tipo de marcadores biológicos tienen valor pronóstico en la supervivencia global y la respuesta al tratamiento en el cáncer de mama, a partir del descubrimiento de metástasis pleurales. 2. Examinar la influencia de la aplicación de talco intrapleural. 3. Establecer un esquema de abordaje terapéutico en base a los diferentes factores pronósticos. Pacientes y métodos: estudiamos una serie de 126 pacientes con cáncer de mama y afectación pleural metastásica. Se examinaron exhaustivamente los factores clásicos del cáncer de mama. Así mismo se calcularon el intervalo libre de enfermedad (ILE), el intervalo entre la aparición del derrame pleural y el abordaje toracoscópico del mismo, el intervalo entre la realización de la pleurodesis con talco y el exitus y el intervalo desde el diagnóstico del tumor primario hasta el exitus. Los factores biológicos (polimorfismos)fueron estudiados a partir del ADN de muestras de sangre periférica o líquido pleural crio conservado. Resultados: el derrame pleural era en el 77% de los casos la primera manifestación de recidiva de la enfermedad. El intervalo libre de enfermedad (ILE) fue de 57,9 meses (13,6-83,3). La supervivencia(expresada como mediana) desde el momento del diagnóstico del tumor primario fue de 77,5 meses (0,83-384). La media de seguimiento tras el talcaje fue de 14,6 meses. Encontramos correlación entre la edad temprana de presentación del cáncer de mama (..) (AU)

Objective: 1. To study what type of biological markers have prognostic value in the global survival rate and response to treatment of breast cancer, regarding discovery of pleural metastases. 2. To examine the influence of the application of intrapleural talc. 3. To establish a therapeutic approach outline based on the different prognosticfactors. Patients and methods: we studied a series of 126 patients with breast cancer and pleural metastasis. The classic factors of breast cancer were thoroughly examined. Also, we calculated the disease-free interval(DFI), the interval between the appearance of the pleural effusion and its thoracoscopic approach, the interval between pleurodesis with talc and death and the interval between the diagnosis of the first tumour and death. The biological factors (polymorphisms) were studied starting with the DNA of peripheral blood samples or cryopreservation of pleural liquid samples. Results: the pleural effusion was the first manifestation of a relapse of the disease in 77% of the cases. The disease-free interval (DFI) was57.9 months (13.6-83.3). The survival rate (expressed as median)from the diagnosis of the first tumour was 77.5 months (0.83-384).The average follow-up after the application of talc was 14.6 months. We found a correlation between the appearance of breast cancer(p=0.003) at a young age and the presence of the A2/A2 allele(homozygote of the CYP-17).Conclusions: the ideal profile for candidates for pleurodesis is those patients with an age >50 years, long DFI, short interval of time between the appearance of the pleural effusion and the application oftalc, absence of other metastasis at the time of thoracoscopy, positive ER (estrogen receptors) and PR (progesterone receptors) and a percentage of lymph nodes infiltrated/extracted less than 50%. The presence of mutated GSTM1 will be related with a more effective (..) (AU)

Importancia de los polimorfismos de la ciclooxigenasa-2 en la presentación clínica de la sarcoidosis / Importance of cyclooxigenase-2 polymorphismsin the clinical presentation of sarcoidosis

OBJETIVO: Estudiar la posible relación entre las manifestaciones clínicas de la sarcoidosis y los polimorfismos del gen de laciclooxigenasa-2 (COX-2).MÉTODO: Estudio multicéntrico observacional transversal en el que participaron 7 hospitales de España. Se incluyeron pacientes diagnosticados de sarcoidosis según criterios internacionales. De cada caso se recogió edad, sexo, método diagnóstico, enzima convertidora de angiotensina, pruebas de función respiratoria, estadio radiológico y clínica del paciente en el momento del diagnóstico. Los hallazgos clínicos se agruparon en respiratorios y sistémicos. Los estudios genéticos se realizaron a partir del ADN obtenido de linfocitos de sangre periférica. El ADN se amplificó mediante PCR convencional y los polimorfismos fueron analizados por sondas de hibridación fluorescentes y curvas de disociación. Se determinaron4 variantes alélicas del gen de la COX-2: COX2.5909T>G,COX2.8473T>C, COX2.926G>C y COX2.3050G>C. RESULTADOS: La muestra se compuso de 131 casos de sarcoidosis (63 hombres; edad: 47 ± 15 años), todos con diagnóstico histológico menos 5 casos. El polimorfismo COX2.3050G>C en homocigosis resultó estar significativamente presente entre los pacientes con manifestaciones sistémicas frente al resto de pacientes (4,6% vs 0%;p=0,045). La presencia de manifestaciones sistémicas de la enfermedad estuvo significativamente asociada a los pacientes portadores del alelo C de dicho polimorfismo (34,4% vs. 18,6%; p=0,031; OR:2,3; IC 95%: 1,03-5,12). El resto de polimorfismos estudiados no estuvieron relacionados con la expresión clínica de la enfermedad. CONCLUSIÓN: La presencia de manifestaciones sistémicas parece estar relacionada con los portadores del alelo C del polimorfismoCOX2.3050G>C de la COX-2 (AU)

OBJECTIVE: To study clinical manifestations of sarcoidosis according to cyclooxigenase-2 (COX-2) polymorphisms. METHOD: Observational cross-sectional multicentre trial in which 7 Spanish hospitals participated. Patients diagnosed withs arcoidosis according to international criteria were included. Age, gender, diagnostic method, angiontens in converting enzyme, pulmonary function tests, radiological stage and clinical findings at the moment of the diagnosis were recorded for each case included. Clinical findings were grouped as respiratory or systemic. Genetic studies were performed on DNA extracted from peripheral blood lymphocytes. DNA was amplified by conventional PCR and polymorphisms were studied by Fluorescent Hybridization Probe-Melting Curves. COX-2 polymorphisms genotyped were COX2.5909T>G, COX2.8473 T>C, COX2.926 G>C y COX2.3050 G>C.RESULTS: 131 sarcoidosis patients (63 males, age: 47 ± 15years) were included. All included patients had a histological diagnosis except for 5 patients. COX2.3050G>C homozygote polymorphism resulted to be significantly present in patients with a systemic manifestation of the disease as compared with the rest of the sample(4,6% vs 0%; p = 0,045). Systemic manifestations were significantly associated with allele C carriers of this polymorphism (34.4% vs.18.6%; p = 0.031; OR: 2.3; IC 95%: 1.03 – 5.12). The rest of the studied polymorphisms were not significantly related to the clinical manifestations of the disease. CONCLUSION: Our results suggest that allele C carriers ofCOX2.3050G>C polymorphism are associated with the systemic manifestations of sarcoidosis (AU)

Association of the 3050G>C polymorphism in the cyclooxygenase 2 gene with systemic sarcoidosis.

BACKGROUND: We investigated the potential association between cyclooxygenase-2 (COX-2) gene polymorphisms and clinical manifestations of sarcoidosis. METHODS: This observational cross-sectional study involved seven hospitals in Spain. We diagnosed patients with sarcoidosis according to the International Criteria. The following variables were recorded: age, gender, initial diagnostic methods, serum angiotensin-converting enzyme (ACE) levels, pulmonary function tests, radiological stage, and clinical findings at diagnosis. Manifestations of sarcoidosis were classified as systemic vs. nonsystemic. Genotyping of four COX-2 polymorphisms (COX2.5909T>G, COX2.8473T>C, COX2.926G>C, and COX2.3050G>C) was undertaken on DNA extracted from peripheral blood lymphocytes using fluorescent hybridization probes and melting curves. RESULTS: A total of 131 sarcoid patients (63 males, mean age: 47 +/- 15 years) were studied. One hundred twenty-six of these patients had one or more positive biopsies. The results demonstrated that genotype distribution for the COX2.3050G>C polymorphism was significantly different between patients with systemic sarcoidosis and those with nonsystemic forms (p = 0.046). After adjustment for age, gender, and serum ACE levels, a significant association between the carriage of at least one C allele of the COX2.3050G>C polymorphism and systemic sarcoidosis was observed (odds ratio [OR]: 2.3; 95% confidence interval [CI]: 1.03-5.12, p = 0.031). Other polymorphisms were not associated with either clinical manifestations of the disease or serum ACE levels. CONCLUSIONS: Our results indicate for the first time that the C allele of the COX2.3050G>C polymorphism is associated with systemic sarcoidosis.

Análisis morfométrico y mineralógico de 14 muestras de talco usado para pleurodesis en distintos países de europayamérica* / Morphometric and mineralogical análisis of 14 samples of talc used for pleurodesis in several european and american countries

FUNDAMENTO DEL ESTUDIO: A pesar de que el talco es elagente más utilizado para pleurodesis, hay controversia respecto asu uso a raíz de la publicación de algunas complicaciones relacionadascon el mismo. Bajo la hipótesis de que diferencias en tamañoy composición pudieran asociarse a distinta incidencia de complicaciones,hemos investigado las características físico-químicas demuestras de talco procedentes de varios países de Europa y América.MÉTODOS: Hemos llevado a cabo un estudio morfométrico ymineralógico de 14 talcos diferentes (9 de Brasil, 3 de Francia, 1de España y 1 de USA). El estudio morfométrico se hizo mediantefotografía de microscopía óptica por dos observadores independientesde nuestro grupo, y mediante microscopía electrónica debarrido, y la composición químico-mineralógica se estudiómediante difracción de rayos X y fluorescencia (Instituto de Cienciade Materiales de Sevilla).RESULTADOS: El diámetro menor de las partículas oscilóentre 3,3 μm (Brasil-4) y 18,5 μm (Brasil-2). El porcentaje de partículasmenores de 10 μm osciló entre 10% (Steritalc® aerosol) y97% (Brasil-4). Respecto a la composición química, encontramostalco como componente mayoritario en ocho muestras (Brasil-2, 6y 7, Francia, España y USA); en el resto el componente mayoritariofue dolomita o flogopita, y en menor porcentaje cuarzo y calcita.CONCLUSIÓN: Observamos gran diversidad en cuanto amorfometría y composición química entre las distintas muestras detalco estudiadas; esto nos lleva a recomendar un estricto análisisdel agente esclerosante, no sólo en relación al tamaño de sus partículassino también en relación a su composición química

AIMS OF THE STUDY: Despite talc being the most commonlysclerosant used for pleurodesis, the ocassional reports ofcomplications occurred after its intrapleural instillation have provokedsome controversy. We hypothezised that differences in particlesize and composition might be associated to incidence of complications,and subsequently investigated the physico-chemicalcharacteristics of talc samples from several European and Americancountries.METHODS: A morphometric and mineralogical study wascarried out on 14 samples of talc obtained from different countries(nine from Brazil, three from France, one from Spain and onefrom the U.S.). Morphometry was performed separately by twoindependent observers through computerized image analysis ofphotographs taken by using optical microscopy under polarizedlight. Scan electron microscopy was used also, and X-rays difractionand fluorescence was used for chemical analysis (Instituto deCiencia de Materiales at Sevilla, Spain).RESULTS: The average minor particle diameter ranged from3,3 μm (Brazilian talc #4) and 18,5 μm (Brazilian talc #2). The percentageof particles smaller than 10 μm ranged between 10% (Steritalc® spray French talc) and 97% (Brazilian talc #4). Regardingchemical composition, we found true talc as the main componentonly in eight samples (Brazilian #2, 6 and 7, plus all French, Spanishand North American samples). Dolomite or flogopite were themain components in the remaining samples, and quartz and calcitewere also found as minoritary components in some cases.Conclusion: We have observed wide differences regardingmorphometry and chemical composition of the talcs studied. Therefore,a strict control of this sclerosant agent is advisable, not onlyregarding the size of particles, but also its chemical composition