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.

Aberrant gene methylation and bronchial dysplasia in high risk lung cancer patients.

INTRODUCTION: The risk for lung cancer is incremented in high degree dysplasia (HGD) and in subjects with hypermethylation of multiple genes. We sought to establish the association between them, as well as to analyze the DNA aberrant methylation in sputum and in bronchial washings (BW). METHODS: Cross sectional study of high risk patients for lung cancer in whom induced sputum and autofluorescence bronchoscopy were performed. The molecular analysis was determined on DAPK1, RASSF1A and p16 genes using Methylation-specific PCR. RESULTS: A total of 128 patients were enrolled in the study. Dysplasia lesions were found in 79 patients (61.7%) and high grade dysplasia in 20 (15.6%). Ninety eight patients out of 128 underwent molecular analysis. Methylation was observed in bronchial secretions (sputum or BW) in 60 patients (61.2%), 51 of them (52%) for DAPK1, in 20 (20.4%) for p16 and in three (3.1%) for RASSF1A. Methylated genes only found in sputum accounted for 38.3% and only in BW in 41.7%, and in both 20.0%. In the 11.2% of the patients studied, HGD and a hypermethylated gene were present, while for the 55.1% of the sample only one of both was detected and for the rest of the subjects (33.6%), none of the risk factors were observed. CONCLUSIONS: Our data determines DNA aberrant methylation panel in bronchial secretions is present in a 61.2% and HGD is found in 15.6%. Although both parameters have previously been identified as risk factors for lung cancer, the current study does not find a significative association between them. The study also highlights the importance of BW as a complementary sample to induced sputum when analyzing gene aberrant methylation.

[Calcific myonecrosis of the leg]. / Mionecrosis calcificante de la pierna.

Calcific myonecrosis is a rare post-traumatic sequela almost exclusively located in the lower extremity, which can be mistaken for an aggressive primary neoplasm. This lesion, initially described by Gallei and Thompson in 1960, is characterized by the formation of a calcified mass that appears decades after trauma. The pathophysiologic mechanism is not fully understood, although the lesion most likely results from post-traumatic ischemia and it may be associated with a common peroneal nerve injury. The typical radiographic image is a fusiform soft tissue mass with linear calcifications. The treatment of choice is conservative in asymptomatic patients because the surgical treatment has a high complication rate. We report four cases of calcific myonecrosis treated surgically in our hospital. Three of the cases had an infection as a complication that required subsequent debridement and special therapies to achieve the resolution of the cases.

Mionecrosis calcificante de la pierna / Calcific myonecrosis of the leg

La mionecrosis calcificante es una rara secuela postraumática que se localiza casi exclusivamente en la extremidad inferior, y que puede ser confundida con una neoplasia primaria agresiva. Esta lesión, descrita inicialmente por Gallei y Thompson en 1960, se caracteriza por la formación de una masa calcificada que aparece varias décadas después de un traumatismo. El mecanismo fisiopatológico no es conocido, sin embargo la lesión parece que es debida a una isquemia postraumática y puede asociarse con una lesión del ciático poplíteo externo. La imagen radiográfica típica es una masa de partes blandas fusiforme con calcificaciones lineales. El tratamiento de elección es conservador en los casos asintomáticos ya que el tratamiento quirúrgico tiene un alto porcentaje de complicaciones. Presentamos 4 casos de mionecrosis calcificante tratados quirúrgicamente en nuestro hospital. Tres de los casos se infectaron por lo que precisaron sendos desbridamientos y terapias especiales para su resolución definitiva (AU)

Calcific myonecrosis is a rare post-traumatic sequela almost exclusively located in the lower extremity, which can be mistaken for an aggressive primary neoplasm. This lesion, initially described by Gallei and Thompson in 1960, is characterized by the formation of a calcified mass that appears decades after trauma. The pathophysiologic mechanism is not fully understood, although the lesion most likely results from post-traumatic ischemia and it may be associated with a common peroneal nerve injury. The typical radiographic image is a fusiform soft tissue mass with linear calcifications. The treatment of choice is conservative in asymptomatic patients because the surgical treatment has a high complication rate. We report four cases of calcific myonecrosis treated surgically in our hospital. Three of the cases had an infection as a complication that required subsequent debridement and special therapies to achieve the resolution of the cases (AU)

Can leptomeningeal myelomatosis be predicted in patients with IgD multiple myeloma?

Involvement of the central nervous system (CNS) by multiple myeloma (MM) is rare. Immunoglobulin D (IgD) MM represents only 1% to 2% of all MM patients. Previous reports show a disproportionate number of patients with IgD MM with leptomeningeal myelomatosis (LMM). Several biological markers have been associated with LMM. The development of LMM in a woman with IgD MM is reported. Our patient should be considered as having a high-risk of CNS disease based on: (i) presence of IgD-lambda MM; (ii) high myeloma burden (stage III); (iii) additional extramedullary disease; (iv) presence of circulating plasma cells, some with plasmablastic morphology; and (v) CD56-negative immunophenotype. The association between these features of MM reported previously and a high risk of LMM is reviewed. Studies including patients with these features are warranted to confirm their attributed LMM risk and to investigate the role of prophylactic chemoradiotherapy in this clinical setting.

Single and repeated bleomycin intratracheal instillations lead to different biomechanical changes in lung tissue.

Single dose of bleomycin induces acute alveolitis followed by a reparative process whilst a repeated dose results in progressive fibrosis, which may lead to distinct lung tissue biomechanical changes. To test this hypothesis, rats were intratracheally instilled with saline (N=11) or bleomycin (2.5U/kg) once (SD, N=8) or three times (RD, N=9) one week apart, and sacrificed 28 days after challenge. Forced oscillatory mechanics as well as the amount of collagen fibre and myeloperoxidase content (MPO(L)) were studied in lung tissue strips. Both elastic modulus (H), tissue damping (G), and MPO(L) increased only in RD-challenged rats. Although fibroblast focus was found in RD, collagen fibre content increased in both challenged groups. However, the amount of collagen fibre in SD group was not enough to induce lung tissue mechanical changes. In conclusion, repeated doses of bleomycin induce inflammatory and fibrogenic behaviour with biomechanical changes mimicking interstitial lung disease in humans.

Linfadenitis necrosante de Kikuchi-Fujimoto y su asociación con el lupus eritematoso sistémico: descripción de 4 nuevos casos / Kikuchi-Fujimoto necrotizing lymphadenitis and its association with systemic lupus erythematosus: description of 4 new cases

Objetivo: Describir las características clínicas de la linfadenitis necrosante de Kikuchi Fujimoto y evaluar su posible asociación con el lupus eritematoso sistémico. Métodos: Diseño retrospectivo. Se revisaron las historias clínicas de aquellos casos de linfadenitis de Kikuchi Fujimoto y lupus eritematoso sistémico que habían sido visitados en la Sección de Reumatología del Hospital Universitari Germans Trias i Pujol durante los años 1984-2001. Resultados: Se identificaron 4 casos de linfadenitis de Kikuchi: 4 mujeres (media de edad, 31 años; límites, 27-34). En todos ellos se llegó al diagnóstico a partir de la biopsia ganglionar. Tres de los casos estaban asociados a lupus eritematoso sistémico. Conclusión: La linfadenitis necrosante de Kikuchi Fujimoto parece estar asociada con el lupus eritematoso sistémico. (AU)

Proliferating trichilemmal tumour: p53 immunoreactivity in association with p27Kip1 over-expression indicates a low-grade carcinoma profile.

AIMS: Alterations of cell-cycle regulatory molecules in tumorigenesis may predict the biological behaviour of neoplasms and greatly contribute to their proper classification. Since the behaviour of proliferating trichilemmal tumour (PTT) is controversial, we decided to explore the possible significance of altered p53 and p27Kip1 immunohistochemical expression patterns in PTT. METHODS AND RESULTS: We evaluated the percentage and distribution of positive tumour cells and compared the results with those obtained from usual trichilemmal cysts (TC) and squamous cell carcinomas with trichilemmal differentiation (SCCT). PTT showed p53 immunoreactivity (50.4 +/- 29.6, mean +/- standard deviation) that was not statistically different from that seen in SCCT (75.2 +/- 36.3). On the other hand, p53 immunostaining was virtually absent in TC cases (positivity for p53 was observed in only one instance in < 1% of cells). As for p27Kip1, the mean percentage of positive cells in PTT (82.7 +/- 9.9) was slightly lower than in TC (90.6 +/- 4.6) but significantly higher than in SCCT (53.4 +/- 30). CONCLUSIONS: The similar p53 immunoreactivity in both PTT and SCCT favour the interpretation of the former as carcinoma, albeit one whose behaviour would be tempered by the well-known regulatory effect exerted by p27Kip1 on the cell cycle.

Towards a 'human-like' model of tuberculosis: intranasal inoculation of LPS induces intragranulomatous lung necrosis in mice infected aerogenically with Mycobacterium tuberculosis.

It is well known that one of the differences between murine and human tuberculosis is the lack of intragranulomatous necrosis in the former. The aim of this study was to create a feasible and reproducible model of an experimental model of murine tuberculosis in which this necrosis should be present. Considering the Shwartzman reaction as a possible explanation for intragranulomatous necrosis in human tuberculosis, C57Bl/6 mice, infected aerogenically with a virulent strain of Mycobacterium tuberculosis, were intranasally inoculated with lipopolysaccharide (LPS) on day 19 postinfection (p.i.). Twenty-four hours later, neutrophils infiltrated the lung parenchyma in a significant level, and 10 days after necrosis could be detected in the centres of primary granulomas, that showed scanty macrophages and large amounts of collagen on an eosinophilic background. On the other hand, a significant decrease in the concentration of colony forming units (CFU) could be appreciated 24 h after the LPS inoculation. Afterwards, nonbronchogenic spreading of granulomas increased and higher levels of interferon (IFN)-gamma mRNA were detected. These results lend support to the Shwartzman reaction as the origin of the intragranulomatous necrosis in the M. tuberculosis infection, and provides a useful tool to improve experimental murine models in tuberculosis.

Palisading and verocay body-prominent dermatofibrosarcoma protuberans: a report of three cases.

UNLABELLED: The aim of this report is to draw attention to nuclear palisading and Verocay body formation as peculiar, previously undescribed histological findings in rare instances of dermatofibrosarcoma protuberans (DFSP). METHODS AND RESULTS: Three indurated, nodular or plaque skin lesions were diagnosed as DFSP on the basis of their storiform proliferation of spindle-shaped cells diffusely infiltrating the dermis and subcutaneous tissue. Sclerosing and giant cell areas were also identified. Unexpectedly, conspicuous nuclear palisading was also noted in all cases and Verocay body formation was present in two. Immunostains were positive for CD34 and negative for S100 protein in every instance. Proliferating cells were seen to display fibroblast-like features by ultrastructural study of one case. CONCLUSIONS: DFSP may rarely show a schwannoid histological appearance as the result of nuclear palisading and even Verocay body formation. In this setting, both the search for DFSP characteristic morphologic features and the performance of CD34 and S100 protein immunohistochemistry will facilitate the correct diagnosis.

Evolution of granulomas in lungs of mice infected aerogenically with Mycobacterium tuberculosis.

Aerogenous infection of C57Bl/6 mice with a virulent strain of Mycobacterium tuberculosis (CL 511) leads to the formation of primary granulomas in the lung where neutrophils, macrophages and subsequently, lymphocytes accumulate progressively around an initial cluster of infected macrophages. The spread of infection through the lung parenchyma gives rise to secondary granulomas featuring numerous lymphocytes that surround a small number of infected macrophages. Afterwards, foamy macrophages add an outer layer to the granulomas, which characteristically respect the pulmonary interstitium and remain confined within the alveolar spaces. This feature, in conjunction with the constant presence of M. tuberculosis in the products of broncho-alveolar lavage, suggests that the upward bronchial migration of infected macrophages may contribute significantly to pulmonary dissemination of mycobacterial infection. The latter would be in agreement with the persistence of chronic pulmonary infection in spite of a concomitant strong T helper 1 cell response.

Varón de 38 años con anemia perniciosa y formaciones polipoides gástricas / Thirty-eight-year-old male with pernicious anemia and gastric polyps

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