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Alberto Ricci,
Claudia De Vitis,
Alessia Noto,
Luigi Fattore, Salvatore Mariotta,
Emanuela Cherubini,
Giuseppe Roscilli,
Giuseppina Liguori,
Giosuè Scognamiglio,
Gaetano Rocco,
Gerardo Botti,
Enrico Giarnieri,
Maria Rosaria Giovagnoli,
Giorgio De Toma,
Gennaro Ciliberto,
Rita Mancini
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ABSTRACT: Lung cancer is the leading cause of cancer-related mortality worldwide. Recent evidence indicates that tumors contain a subpopulation of cancer stem cells (CSCs) that are responsible for tumor maintenance and spread. CSCs have recently been linked to the occurrence of epithelial-to-mesenchymal transition (EMT). Neurotrophins (NTs) are growth factors that regulate the biology of embryonic stem cells and cancer cells, but still little is known about the role NTs in the progression of lung cancer. In this work, we investigated the role of the NTs and their receptors using as a study system primary cell cultures derived from malignant pleural effusions (MPEs) of patients with adenocarcinoma of the lung. We assessed the expression of NTs and their receptors in MPE-derived adherent cultures vs. spheroids enriched in CSC markers. We observed in spheroids a selectively enhanced expression of TrkB, both at the mRNA and protein levels. Both K252a, a known inhibitor of Trk activity, and a siRNA against TrkB strongly affected spheroid morphology, induced anoikis and decreased spheroid forming efficiency. Treatment with neurotrophins reversed the inhibitory effect of K252a. Importantly, TrkB inhibition caused loss of vimentin expression as well as that of a set of transcription factors known to be linked to EMT. These ex vivo results nicely correlated with an inverse relationship between TrkB and E-cadherin expression measured by immunohistochemistry in a panel of lung adenocarcinoma samples. We conclude that TrkB is involved in full acquisition of EMT in lung cancer, and that its inhibition results in a less aggressive phenotype.
Cell cycle (Georgetown, Tex.) 05/2013; 12(11). · 5.36 Impact Factor
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Damiano Magrì,
Piergiuseppe Agostoni,
Agnese Ricotta,
Lara Pisani,
Filippo Maria Cauti,
Alessandro Onofri,
Pierdonato Bruno,
Alberto Ricci,
Massimo Volpe,
Simona Marchitti, Salvatore Mariotta,
Speranza Rubattu
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ABSTRACT: BACKGROUND: Lung diffusion for carbon monoxide (DL(CO)) has been shown to associate with the risk of pulmonary arterial hypertension development and, most likely, with right ventricular (RV) myocardial dysfunction in sarcoidosis patients. Besides its known role as a marker of left ventricular dysfunction, experimental evidence suggests a role of NT-proAtrial Natriuretic Peptide (NT-proANP) also in modulating pulmonary circulation. We therefore investigated possible relationships between NT-proANP, lung diffusion impairment and RV dysfunction. METHODS: Thirty-two pulmonary sarcoidosis outpatients and eighteen volunteers underwent full clinical assessment, including full lung function tests and Doppler echocardiography integrated with tissue Doppler imaging (TDI) study. Resting circulating NT-proBNP and NT-proANP plasma levels were also determined. RESULTS: NT-proANP and RV-myocardial performance index (RV-MPI) were significantly higher in those patients with the greatest DL(CO) impairment, whereas no differences were found for NT-proBNP values. At multivariable analysis, only DL(CO) (β: -0.496; standard error: 3.38; p=0.000) and RV-MPI (β: 0.373; standard error: 6.56; p=0.031) remained significantly associated with NT-proANP levels. CONCLUSIONS: Our finding may support a key role of NT-proANP in the complex mechanisms underlying modulation of lung function. An early increase in pulmonary vascular resistance may stimulate NT-proANP increase, thus explaining its association with signs of early RV myocardial dysfunction. This hypothesis warrants further confirmation.
European Journal of Internal Medicine 01/2013; · 2.00 Impact Factor
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Enrico Giarnieri,
Claudia De Vitis,
Alessia Noto,
Giuseppe Roscilli,
Gerardo Salerno, Salvatore Mariotta,
Alberto Ricci,
Bruno Pierdonato,
Giuseppe Russo,
Andrea Laurenzi,
Maria Rosaria Giovagnoli,
Gennaro Ciliberto,
Rita Mancini
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ABSTRACT: Epithelial-to-Mesenchymal Transition (EMT) is a process in which cells undergo a developmental switch from epithelial to mesenchymal phenotype. This process has been related to embryologic morphogenesis but also to cancer progression and metastasis. The aim of the current study was to investigate the expression of EMT related markers in adherent and spheroid cell cultures derived from Malignant Pleural Effusions (MPEs) of patients affected by lung adenocarcinoma. On the basis of efficient in vitro propagation, six cases of MPEs were selected and analyzed by immunocytochemistry staining for EMT markers and by RT-PCR for transcription factors known to orchestrate EMT. EMT markers immunostaining showed in spheroids a statistically significant correlation between the loss of E-cadherin immunoreactivity and overexpression of N-cadherin (P < 0.001). Likewise loss of EpCAM epithelial marker was coincident with Vimentin overexpression (P < 0.001). RT-PCR analysis of transcription factors Snail, Slug and Twist showed a highly variable expression, although a general trend to increase was observed. Importantly in some selected cases it was possible to establish a precise relationship between spheroid formation, EMT switch and increased upregulation of the marker related to cancer stemness such as ALDH positivity. Therefore MPE-derived cell cultures, while recapitulating the heterogeneity of lung cancer, are a suitable system to study the mechanisms at the basis of EMT and to understand its relationship with the generation of cancer stem cells. J. Cell. Physiol. © 2012 Wiley Periodicals, Inc.
Journal of Cellular Physiology 12/2012; · 3.87 Impact Factor
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Alberto Ricci,
Emanuela Cherubini,
Davide Scozzi,
Vittorio Pietrangeli,
Luca Tabbì,
Salvatore Raffa,
Laura Leone,
Vincenzo Visco,
Maria Rosaria Torrisi,
Pierdonato Bruno,
Rita Mancini,
Gennaro Ciliberto,
Claudio Terzano, Salvatore Mariotta
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ABSTRACT: Autophagy is the main cellular pathway for degradation of long-lived proteins and organelles and regulates cell fate in response to stress. Beclin 1 is a key regulator of this process. In some settings autophagy and apoptosis seems to be interconnected. Recent reports indicate that fibroblasts in idiopathic pulmonary fibrosis (IPF) acquire resistance to apoptosis. Here we examined the expression of beclin 1, and of the anti apoptotic protein Bcl-2 in human IPF fibroblasts using immunohistochemistry and molecular biology in both bioptic sections, in primary cultured of fibroblasts taken from patients with IPF and in fibroblast cell lines. Expression of beclin 1 and in fibroblasts from IPF was down-regulated in comparison with fibroblasts from normal lungs while the anti-apoptotic protein Bcl-2 expression was over-expressed. Treatment of fibroblasts cell cultures with cisplatin induced a significant increase in beclin 1, and caspase 3 protein levels but a reduction in Bcl-2 expression. These observations were confirmed by the analysis of acid compartments and transmission electron microscopy. Our results demonstrate a modified expression of the apoptotic beclin1 and of the oncogene Bcl-2 proteins in human IPF fibroblasts suggesting the existence of an autophagy/apoptosis system dysfunction. J. Cell. Physiol. © 2012 Wiley Periodicals, Inc.
Journal of Cellular Physiology 12/2012; · 3.87 Impact Factor
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Pierdonato Bruno,
Giovanna Gentile,
Rita Mancini,
Claudia De Vitis,
Maria Cristina Esposito,
Davide Scozzi,
Mario Mastrangelo,
Alberto Ricci,
Ibrahim Mohsen,
Gennaro Ciliberto,
Maurizio Simmaco, Salvatore Mariotta
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ABSTRACT: CpG island hypermethylation of gene promoters and regulatory regions is a well-known mechanism of epigenetic silencing of tumor suppressors and is directly linked to carcinogenesis. Wilm's tumor gene (WT1) is a tumor suppressor protein involved in the regulation of human cell growth and differentiation and a modulator of oncogenic K Ras signaling in lung cancer. Changes in the pattern of methylation of the WT1 gene have not yet been studied in detail in human lung cancer. In this study we compared the methylation profile of WT1 gene in samples of neoplastic and non-neoplastic lung tissue taken from the same patients.
DNA was extracted from neoplastic and normal lung tissue obtained from 16 patients with non small cell lung cancer (NSCLC). The methylation status of 29 CpG islands in the 5' region of WT1 was determined by pyrosequencing. Statistical analysis was carried out by T test and Mann Whitney test.
The mean percentage of methylation, considering all CpG islands of WT1 in the neoplastic tissues of the 16 NSCLC patients, was 16.2±3.4, whereas in the normal lung tissue from the same patients it was 5.6±1.7 (p<0.001). Adenocarcinomas presented higher methylation levels than squamous cell carcinomas (p<0,001).
Methylation of WT1 gene is significantly increased in NSCLC. Both histotype and exposure to cigarette smoke heavily influence the pattern of CpG islands which undergo hypermethylation.
Biochemical and Biophysical Research Communications 08/2012; 426(3):306-9. · 2.48 Impact Factor
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Alberto Ricci,
Emanuela Cherubini,
Alessandra Ulivieri,
Luca Lavra,
Salvatore Sciacchitano,
Davide Scozzi,
Rita Mancini,
Gennaro Ciliberto,
Armando Bartolazzi,
Pierdonato Bruno,
Paolo Graziano, Salvatore Mariotta
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ABSTRACT: Homeodomain-interacting protein kinase 2 (Hipk2) is an emerging player in cell response to genotoxic agents that contributes to the cell's decision between cell cycle arrest or apoptosis. HIPK2 acts as co-regulator of an increasing number of transcription factors and modulates many different basic cellular processes such as apoptosis, proliferation, DNA damage response, differentiation. Idiopathic pulmonary fibrosis (IPF) is characterized by an anatomical disarrangement of the lung due to fibroblast proliferation, extracellular matrix deposition and lung function impairment. Although the role of inflammation is still debated, attention has been focused on lung cell functions as fibroblast phenotype and activity. Aim of the present study was to analyze the loss of heterozygosity (LOH) at HIPK2 locus 7q32.34 in human lung fibroblasts and the HIPK2 expression in 15 IPF samples and in four primary fibroblast cell cultures isolated from IPF biopsies using semi-quantitative RT-PCR, Western blots and immunohistochemistry. We demonstrated a frequency of LOH in IPF fibroblasts of 46% for the internal D7S6440 microsatellite and 26.6% for the external D7S2468 microsatellite. Furthermore, we demonstrated low HIPK2 protein expression in those fibroblasts from IPF patients that present the HIPK2 LOH. The restoration of HIPK2 expression in IPF derived cells induced a significant reduction of chemoresistance after treatment with cisplatin. The results obtained allow us to hypothesize that HIPK2 dysfunction may play a role in fibroblasts behavior and in IPF pathogenesis. HIPK2 may be considered as a novel potential target for anti-fibrosis therapy. J. Cell. Physiol. 228: 235-241, 2013. © 2012 Wiley Periodicals, Inc.
Journal of Cellular Physiology 06/2012; 228(1):235-41. · 3.87 Impact Factor
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ABSTRACT: The results of a recent study have shown the superiority of treatment with gefitinib or erlotinib in lung tumors positive for epidermal growth factor receptor (EGFR) mutation. As a consequence, the complete diagnosis of lung cancer cannot be limited to histotype classification, but should include a series of molecular biology analyses. In most cases, the diagnosis of lung cancer is performed on cytological specimens; therefore, there is a need to obtain a complete and reliable molecular diagnosis on cytologic specimens. Brushing, transbronchial needle aspiration (TBNA) and broncho alveolar lavage during fibro-bronchoscopy allow the sampling of the lung and the mediastinal lymph node. The aim of this study was to demonstrate that direct sequencing of exons 19 and 21 of EGFR in lung tumors, carried out on the cytological samples obtained through fibro-bronchoscopy, is as reliable as the same analysis carried out on a histological surgical sample obtained from the same individual. We considered 50 patients with a histological diagnosis of lung adenocarcinoma whose cytological samples, obtained by fibro-bronchoscopy and histological samples, obtained by surgical resection were available. A comparison of the sensitivity and reliability of the molecular biology analyses carried out on histological and cytological samples of the same patient was carried out. The combined mutation percentage of exons 19 and 21 of EGFR was 10%. The results of the analyses carried out on cytological samples matched those obtained from the histological samples. The feasibility of EGFR analysis on cytological samples has already been demonstrated in previous studies, however these studies referred to the method of fluorescence in situ hybridization, or did not perform any comparison between histological samples from the same patient; our work, on the other hand, shows that direct sequencing of exons 19 and 21 of the EGFR gene is feasible on fibro-bronchoscopy cytological samples with the same reliability offered by the histological samples obtained from the same patient.
Anticancer research 12/2011; 31(12):4207-10. · 1.73 Impact Factor
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Maurizio Luisetti,
Pierdonato Bruno,
Zamir Kadija,
Takuji Suzuki,
Salvatore Raffa,
Maria Rosaria Torrisi,
Ilaria Campo,
Francesca Mariani,
Ernesto Pozzi,
Bruce C Trapnell, Salvatore Mariotta
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ABSTRACT: Extensive pulmonary fibrosis is a rare occurrence in pulmonary alveolar proteinosis. We report 2 cases that have interesting implications. A female patient was diagnosed with autoimmune pulmonary alveolar proteinosis that evolved over 7 years into diffuse fibrosis. In a male patient with diffuse fibrosis we incidentally detected electron microscopic features of alveolar surfactant accumulation and positive autoantibodies to granulocyte-macrophage colony stimulating factor. In the male patient we speculated that the pulmonary fibrosis might have been preceded by an asymptomatic phase of autoimmune pulmonary alveolar proteinosis, and that we should investigate the involvement of surfactant dysfunction in the pathogenesis of fibrotic lung disease.
Respiratory care 04/2011; 56(10):1608-10. · 2.01 Impact Factor
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Rita Mancini,
Enrico Giarnieri,
Claudia De Vitis,
Donatella Malanga,
Giuseppe Roscilli,
Alessia Noto,
Emanuele Marra,
Carmelo Laudanna,
Pietro Zoppoli,
Pasquale De Luca, [......],
Giuseppe Mesiti,
Luigi Aurisicchio,
Alberto Ricci, Salvatore Mariotta,
Lara Pisani,
Claudio Andreetti,
Giuseppe Viglietto,
Erino A Rendina,
Maria Rosaria Giovagnoli,
Gennaro Ciliberto
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ABSTRACT: Malignant pleural effusions (MPEs) could represent an excellent source to culture a wide variety of cancer cells from different donors. In this study, we set up culture conditions for cancer cells deriving from MPEs of several patients affected by the most frequent form of lung cancer, namely the subset of non small cell lung cancers (NSCLC) classified as Lung Adenocarcinomas (AdenoCa) which account for approximately 40% of lung cancer cases. AdenoCa malignant pleural effusions gave rise to in vitro cultures both in adherent and/or in spheroid conditions in almost all cases analyzed. We characterized in greater detail two samples which showed the most efficient propagation in vitro. In these samples we also compared gene profiles of spheroid vs adherent cultures and identified a set of differentially expressed genes. Finally we achieved efficient tumor engraftment in recipient NOD/SCID mice, also upon inoculation of small number of cells, thus suggesting indirectly the presence of tumor initiating cells.
PLoS ONE 01/2011; 6(7):e21320. · 4.09 Impact Factor
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ABSTRACT: Transbronchial needle aspiration (TBNA) is a bronchoscopic technique allowing the sampling of cytological/histological material from mediastinal lymph nodes. TBNA is routinely used only in few centers for the staging of lung cancer, and even less frequently for the diagnosis of mediastinal metastases from extrapulmonary tumors. We illustrate 5 cases of mediastinal metastases from extrapulmonary tumors observed at our center in order to emphasize the usefulness of cytology and TBNA in the diagnosis of these pathologies. The 5 cases illustrated were: seminoma, uterine cervical carcinoma, pleural mesothelioma, pancreatic carcinoma, pericardial mesothelioma. In these 5 cases, albeit not of lung cancer, the cytology on TBNA allowed the rapid formulation of the correct diagnosis; its main advantage is that it can be performed during a simple fiberbronchoscopy under local anesthesia with less risk and at a lower cost than a computed tomography-guided needle biopsy or mediastinoscopy.
Anticancer research 11/2010; 30(11):4769-72. · 1.73 Impact Factor
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Claudio Terzano,
Vittoria Conti,
Fabio Di Stefano,
Angelo Petroianni,
Daniela Ceccarelli,
Elda Graziani, Salvatore Mariotta,
Alberto Ricci,
Antonio Vitarelli,
Giovanni Puglisi,
Corrado De Vito,
Paolo Villari,
Luigi Allegra
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ABSTRACT: We evaluated comorbidity, hospitalization, and mortality in chronic obstructive pulmonary disease (COPD), with special attention to risk factors for frequent hospitalizations (more than three during the follow-up period), and prognostic factors for death. Two hundred eighty-eight consecutive COPD patients admitted to respiratory medicine wards in four hospitals for acute exacerbation were enrolled from 1999 to 2000 in a prospective longitudinal study, and followed up until December 2007. The Charlson index without age was used to quantify comorbidity. Clinical and biochemical parameters and pulmonary function data were evaluated as potential predictive factors of mortality and hospitalization. FEV(1), RV, PaO(2), and PaCO(2) were used to develop an index of respiratory functional impairment (REFI index). Hypertension was the most common comorbidity (64.2%), followed by chronic renal failure (26.3%), diabetes mellitus (25.3%), and cardiac diseases (22.1%). Main causes of hospitalization were exacerbation of COPD (41.2%) and cardiovascular disease (34.4%). Most of the 56 deaths (19.4%) were due to cardiovascular disease (67.8%). Mortality risk depended on age, current smoking, FEV(1), PaO(2), the REFI index, the presence of cor pulmonale, ischemic heart disease, and lung cancer. Number and length of hospital admissions depended on the degree of dyspnea and REFI index. The correct management of respiratory disease and the implementation of aggressive strategies to prevent or treat comorbidities are necessary for better care of COPD patients.
Beiträge zur Klinik der Tuberkulose 08/2010; 188(4):321-9. · 1.90 Impact Factor
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Alberto Ricci, Salvatore Mariotta,
Elena Pompili,
Rita Mancini,
Elena Bronzetti,
Claudia De Vitis,
Lara Pisani,
Emanuela Cherubini,
Pierdonato Bruno,
Giorgetta Gencarelli,
Maria R Giovagnoli,
Claudio Terzano,
Gennaro Ciliberto,
Enrico Giarnieri,
Lorenzo Fumagalli
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ABSTRACT: Neurotrophins (NTs) expression was assessed in malignant and non-malignant pleural effusions (inflammatory exudates and transudates). Enzyme-linked immunosorbent assay, in malignant exudates from small and non-small cell lung cancer (SCLC and NSCLC), detected nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3), and their levels are higher as compared with inflammatory and transudative effusions. By immunoblots, in cultured cancer cells coming from malignant pleural effusions, NTs and low- and high-affinity NT receptors were detected in a percentage of SCLC and NSCLC. Proliferation assay demonstrated that BDNF significantly increased cancer cell proliferation in vitro, on the contrary, NT-3 reduced cancer cell growth rate and NGF did not modify cell growth. Moreover, NGF protects cells from death during starvation. These effects are reverted by the addition of NT receptor antagonists. Cultured cancer cells injected into the lung of immunodeficient mice generate lung tumors expressing NTs and NT receptors. These findings suggest that NTs may be able to modulate cancer cell behavior and their growth.
Growth factors (Chur, Switzerland) 03/2010; 28(4):221-31. · 2.47 Impact Factor
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ABSTRACT: A 30-year-old woman was admitted to our hospital with severe shortness of breath. A transthoracic echocardiogram showed moderate pericardial effusion with a lesion in the right atrium, confirmed by chest CT scan and cardiac RMN. Pleural and middle lobe involvement occurred within one month. Middle lobe biopsy was performed and pathological examination confirmed the diagnosis of metastatic angiosarcoma. After two months, because of recurrent pleural effusions, chemical pleurodesis was performed. Chemotherapy was started but the patient died four months after the diagnosis. This case highlights the misdiagnosis at initial clinical presentation, available diagnostic approaches and therapeutic options for cardiac angiosarcoma.
Case Reports in Oncology 01/2010; 3(1):24-29.
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ABSTRACT: Abstract Alveolar macrophages play a crucial role in regulating lung immune responses and in maintaining the integrity of the respiratory tract. Neurotrophins (NTs), besides to their neurotrophic activities, exhibit physiological effects in the immune system. In this study, nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), NT-3 and low- (p75) and high affinity (Trks) NT receptors were investigated by immunocytochemistry in cytospin centrifuged preparations of human alveolar macrophages. Approximately 2.5% alveolar macrophages were immunoreactive for NGF, whereas no macrophages displaying immunoreactivity for BDNF or NT-3 were observed. A 3.5% macrophages displayed immunoreactivity for TrkA-receptor protein, 10% for TrkB-receptor protein (full length isoform), and 2% for TrkC-receptor protein. No low-affinity p75 NT and TrkB[-] truncated isoform receptor immunoreactive macrophages were found. These findings support the hypothesis that NTs and the corresponding receptors may play a role in regulating immunological and functional activity of alveolar macrophages via paracrine/autocrine mechanisms.
Growth Factors 07/2009; 18(3):193-202. · 1.65 Impact Factor
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ABSTRACT: The pharmacological profile and the anatomical localization of Ca2+channels of the L-type were investigated in the human pulmonary artery to identify possible mechanisms involved in the regulation of the pulmonary vascular tone. Analysis was performed on slide-mounted frozen sections of human pulmonary artery using radioligand binding assay techniques associated with light microscope autoradiography. [3H]-Nicardipine was used as ligand. Human renal and right coronary arteries also were used as systemic reference arteries. Binding of [3H]-nicardipine to sections of human pulmonary artery was time-, temperature- and concentration-dependent, saturable and reversible. In the human pulmonary artery, the apparent equilibrium dissociation constant (Kd) was 0.12±0.02 nM and the maximum density of binding sites (Bmax) was 38.15±2.25 fmol/mg tissue. Kdvalues were 0.3±0.01 nM and 0.5±0.02 in the human renal artery and right coronary artery respectively. Bmm values were 248 5 16 fmol/mg tissue and 173±9.5 fmol/mg tissue in the human renal artery and right coronary artery respectively. The pharmacological profile of [3H]-nicardipine binding to sections of human pulmonary artery was consistent with the labeling of Ca2+channels of the L-type. It was similar in the pulmonary artery and in the human renal and right coronary arteries. Light microscope autoradiography revealed a high density of [3H]-nicardipine binding sites within smooth muscle of the tunica media of human pulmonary artery as well as of human renal and right coronary arteries. A lower accumulation of the radioligand occurred in the tunica adventitia. No specific binding was noticeable in the tunica intima. Our data suggest that human pulmonary artery expresses Ca2+channels of the L-type sensitive to dihydropyridines. These sites have similar affinity and lower density than those expressed by systemic arteries. The presence of Ca2+channels of the L-type in human pulmonary artery suggests that their pharmacological manipulation may be considered in the treatment of pulmonary hypertension
Clinical and Experimental Hypertension 07/2009; 20(4):389-402. · 1.07 Impact Factor
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Paola Rogliani,
Marco Mura,
Paolo Mattia,
Amedeo Ferlosio,
Gianfranco Farinelli, Salvatore Mariotta,
Paolo Graziano,
Gabriella Pezzuto,
Alberto Ricci,
Cesare Saltini,
Augusto Orlandi
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ABSTRACT: Idiopathic pulmonary fibrosis has been associated with emphysema in cigarette smokers as a new clinical entity: combined pulmonary fibrosis and emphysema (CPFE). In order to compare histomorphometrical, roentgenological and immunohistochemical aspects of usual interstitial pneumonia (UIP) with and without associated pulmonary emphysema, 17 patients with biopsy-proven UIP were evaluated. Morphometrical evaluation of lung parenchyma destruction was used to divide patients in two subgroups: emphysema/UIP (n=9) and UIP alone (n=8); four patients with biopsy-proven emphysema without fibrosis were also evaluated. At HRTC scan, emphysematous lesions were prevalent in the upper fields of both emphysema/UIP and emphysema groups and the distribution of fibrotic lesions was similar in emphysema/UIP compared to UIP alone. The semiquantitative histopathological fibrotic score was also similar in emphysema/UIP and UIP alone. In addition, the expression of tumor necrosis factor (TNF)-alpha, matrix metalloproteinase (MMP)-2, MMP-9, MMP-7 and membrane type 1-metalloproteinase (MT1-MMP) by fibroblasts of myofibroblastic foci was similar in emphysema/UIP and UIP alone patients. In contrast, fibroblasts in areas of parenchymal destruction of emphysema/UIP expressed MMP-2, MMP-9, MMP-7 and MT1-MMP at variable but significantly higher levels when compared to emphysema subjects, in the presence of similar levels of TIMP-1, TIMP-2 and TNF-alpha. Fibrotic and emphysematous lesions in emphysema/UIP patients appear to follow the roentgenological and histopathological patterns expected for either UIP or emphysema. Interstitial fibroblast activation is more pronounced in the areas of lung destruction in emphysema/UIP compared to those with emphysema alone, as for exaggerated tissue remodeling.
Respiratory medicine 09/2008; 102(12):1753-61. · 2.33 Impact Factor
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ABSTRACT: Subjects with bronchial asthmatic symptoms, forced expiratory volume in 1 second (FEV(1)) and FEV(1)/forced vital capacity [FVC] > 80% could show a positive reversibility test with salbutamol in about 25% of the cases. The aim of this study was to evaluate if a limit functional value for spirometry exists where a reversibility test using salbutamol, over this limit, is not necessary to confirm the diagnosis of asthma. Four hundred patients (mean age 31.12 +/- 10.99) with asthmatic symptoms and normal spirometry (mean FEV(1) 96.06 +/- 11.82%, mean FEV(1)/FVC 98.89 +/- 6.03%) were recruited. The number of subjects with reversible airflow obstruction (RAO) was evaluated using the following different criteria of reversibility: FEV(1) and FVC >or= slant 12%, peak expiratory flow (PEF) >or= slant 15%, FEF(25 -75) >or= slant 35%, and at least one of these (ANY). In subjects with baseline FEV(1) > 100% (150 patients), 26 (17.3%) patients showed a FEV(1) increase > 12% and, among the criteria used, 45 patients (30%) showed ANY. In subjects with baseline FEV(1)/FVC > 100% (204 patients), 36 patients (17.6%) showed a FEV(1) increase > 12% and 53 patients (26.8%) showed ANY. In subjects with baseline FEF(25 -75) > 70% (209 patients), 26 (12.44%) and 49 (23.44%) patients, respectively, showed an increase in FEV(1) > 12%, and ANY. In 56 patients with baseline cut-offs (evaluated together) FEV(1) > 100%, FEV(1)/FVC > 100% and FEF(25-75) > 70%, 10 patients (17.85%) showed a RAO with ANY. FEV(1) > 121% or FEV(1)/FVC > 110.8% or FEF(25 -75) > 110% were cut-off values identified in this study where no RAO subjects were found over these limits. In conclusion, baseline FEV(1) > 100%, FEV(1)/FVC > 100% and FEF(25 -75) > 70% cannot be considered cut-offs where it is not necessary to perform a reversibility test with a bronchodilator over these limits. It is improbable to find a positive reversibility test using salbutamol in patients with asthmatic symptoms and FEV(1) > 121% or FEV(1)/FVC > 110.8% or FEF(25 -75) > 110% to confirm the diagnosis of asthma.
Journal of Asthma 08/2008; 45(6):479-83. · 1.52 Impact Factor
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ABSTRACT: Serous effusions are frequently a clinical manifestation of metastatic disease, with lung, breast and ovarian carcinoma and mesothelioma leading the list. The diagnosis of malignant effusion signifies disease progression and is associated with a worsening patient prognosis. The ability to grow in a dense exudative fluid suggests that the malignant cells are capable of acquiring nutrients, surviving and proliferating, despite the lack of a solid-phase scaffold. During proliferation, neoplastic cells release ligands and matrix metalloproteinases (MMPs) into their environment, which dissolve the extracellular matrix (ECM). Tissue inhibitors of metalloproteinase (TIMPs) are endogenous regulators of MMPs, the principal enzymes responsible for the degradation of ECM in metastasis, and reduce their proteolytic activity. TIMP-2 has demonstrated an association between high tumor tissue expression levels and poor prognosis. The purpose of this preliminary study is to investigate, by immunocytochemistry, TIMP-2 expression in non-neoplastic and metastatic adenocarcinoma pleural effusions. We selected 16 cases of reactive mesothelio, 7 of normal mesothelio, 14 of lung adenocarcinoma, 9 from the ovary, 4 from the gastrointestinal tract and 3 from the breast. In 23/30 cases (76%), we detected adenocarcinoma cells with strong TIMP-2 expression. Positive TIMP-2 expression was found in 2/7 cases (28%) of normal and 2/16 (12%) of reactive mesothelio. A statistical association was detected between TIMP-2 expression and metastatic adenocarcinoma cells compared to reactive and normal mesothelial cells (p<0.00003). The calculated sensitivities for TIMP-2 compared to CEA and Ber-EP4 were, respectively, 76.7, 80.0 and 93.3%, and the specificities 82.6, 95.7 and 87.0%. In conclusion, immunocytochemical detection of TIMP-2 could be considered an interesting marker in metastatic adenocarcinoma pleural effusions, and could possibly be used as a component of an antibody panel in diagnostic cytopathology.
Oncology Reports 03/2008; 19(2):483-7. · 1.84 Impact Factor
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ABSTRACT: Parasympathetic nerves provide the dominant autonomic innervation of the upper and lower airways. They release acetylcholine that, activating post-junctional muscarinic receptors, causes bronchoconstriction, mucous secretion and vasodilation. Dysfunction of the upper and lower airways frequently coexist, and they appear to share key elements of pathogenesis.
The present study has assessed the expression and pattern of cholinergic muscarinic receptor subtypes in peripheral blood lymphocytes harvested from allergic rhinitis patients with different degree of bronchial hyperresponsiveness detected by methacholine challenge test.
Radioligand binding assay for determining the density of muscarinic cholinergic receptor subtypes; immunoblot analysis for assessing the characteristic of muscarinic cholinergic receptor subtype protein and immunocytochemical techniques for investigating the cellular localization of receptors.
An increased expression of M2 and M5 receptor proteins was observed in peripheral blood lymphocytes of allergic rhinitis patients in comparison with healthy control individuals. M3 receptor subtype decreased in allergic rhinitis patients with normal or mild responses to methacholine. A trend versus a return to normal value was found in moderate and severe responders. No changes of the M4 receptor subtype were found.
Increase in M2 receptor expression correlated with disease severity and bronchial hyperreactivity. Changes in muscarinic cholinergic receptor expression in allergic rhinitis underline a role of cholinergic system of immune cells in allergic airway disease.
Studies addressed to rhinitis and asthma have identified many similarities. Our results indicate that changes in peripheral blood lymphocyte muscarinic receptor expression may reflect the cholinergic involvement into allergic airway diseases.
Pulmonary Pharmacology & Therapeutics 02/2008; 21(1):79-87. · 2.80 Impact Factor
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ABSTRACT: Idiopathic interstitial pneumoniae (IIPs) are characterized by fibroblast proliferation, extracellular matrix deposition and progressive lung function impairment. Because effective therapeutic strategies still remain limited, research has been directed toward the identification of novel targets for additional therapeutic options. The neurotrophins (NTs) nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and NT-3, beside their importance in nervous, endocrine and immune system activities, participate in chronic inflammatory disorders and in repair processes.
We have investigated NT and high and low affinity NT receptor expression in IIPs using immunoblots and immunohistochemistry. Fourteen idiopatic pulmonary fibrosis/usual interstitial pneumoniae (IPF/UIP), eight non specific pneumoniae (NSIP) and eight respiratory bronchiolitis-associated interstitial lung disease (RB-ILD) were analyzed.
Immunoblots revealed that NT and high affinity NT receptor proteins were more abundantly expressed in IPF/UIP than NSIP and RB-ILD patients. In RB-ILD, a faint expression of NT-3 and NT receptors were detected. NT and NT receptor immunostaining was detected in interstitial cells from IPF/UIP, NSIP and RB-ILD patients by immunohistochemistry. Fibroblastic foci in IPF/UIP strongly stained for BDNF and its high affinity receptor TrkB and in lesser amount for NGF, NT-3 and their respective high affinity receptors TrkA and TrkC. Furthermore, in fibroblast culture derived from IPF/UIP patients, the proliferation rate of primary culture and clones derived from primary lines was stimulated by BDNF but down regulated by NT-3. In contrast, NGF did not influence IPF/UIP fibroblasts proliferation.
Our data suggest that that NTs may exert differential activities on lung fibroblasts and may be considered as potential regulatory molecules influencing fibroblast behavior in IPF/UIP patients. Therefore, NTs may play a role in IIPs patho-physiology representing novel potential therapeutic targets.
Sarcoidosis, vasculitis, and diffuse lung diseases: official journal of WASOG / World Association of Sarcoidosis and Other Granulomatous Disorders 04/2007; 24(1):13-23. · 1.27 Impact Factor
