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Several clinical manifestations attributed to syphilitic infection have been described in the literature. This is important because of the increased incidence of this infection mainly among men who have sex with men. We report the case of an HIV-infected patient who was hospitalized due to severe cholestasis, initially interpreted as drug-induced liver disease. It evolved slowly, with increasing intrahepatic cholangitis confirmed by magnetic resonance cholangiography and the appearance of several pulmonary nodules. In lung and liver biopsies, the presence of Treponema pallidum was detected by molecular and immunohistochemistry techniques, and a diagnosis of cholangitis and pulmonary syphilis was made.

Current guidelines recommend vancomycin and linezolid as first-line agents against methicillin-resistant Staphylococcus aureus (MRSA) nosocomial pneumonia. Telavancin is a potential new therapeutic alternative, specifically in monomicrobial MRSA pneumonia. This study compared the efficacies of telavancin versus linezolid in a porcine model of severe MRSA pneumonia. In 18 mechanically ventilated pigs (32.11 ± 1.18 kg), 75 ml of 10(6) CFU/ml of MRSA was administered into each pulmonary lobe. After the onset of pneumonia, pigs were randomized into three groups: a control group, a group receiving 22.5 mg/kg of body weight every 24 h (q24h) of telavancin, and a group receiving 10 mg/kg q12h of linezolid intravenously. Tracheal aspirate and bronchoalveolar lavage (BAL) fluids were cultured every 24 h. After 48 h of treatment, tissue samples were collected from the ventral and dorsal sections of each lobe. Microbiological and histopathological analyses were performed. Lung tissue concentrations differed among the groups (P = 0.019), with the lowest MRSA lung burden in the telavancin group (P

COVID-19 pandemic has dramatically impacted the health national systems, interfering with the access to standard care of other illnesses. However, the real impact of COVID-19 crisis on the cancer care remains unknown. We assessed the access to invasive procedures and critically ill patient units (intensive and intermediate care) in patients with lung cancer hospitalized during pandemic period (during-Covid) compared to the same period in 2019 (before-Covid).

Long intergenic noncoding RNA-p21 (lincRNA-p21) is a long noncoding RNA transcriptionally activated by tumor protein p53 (TP53) and hypoxia inducible factor 1 alpha subunit (HIF1A). It is involved in the regulation of TP53-dependent apoptosis and the Warburg effect. We have investigated the role of lincRNA-p21 in NSCLC.LincRNA-p21 expression was assessed in tumor and normal tissue from 128 patients with NSCLC and correlated with time to relapse and cancer-specific survival (CSS). H23, H1299, and HCC-44 cell lines were cultured in hypoxic conditions after silencing of lincRNA-p21. The TaqMan human angiogenesis array was used to explore angiogenesis-related gene expression. Levels of the protein vascular endothelial growth factor A were measured by enzyme-linked immunosorbent assay in the cell supernatants. Angiogenic capability was measured by human umbilical vein endothelial cell tube formation assay. Microvascular density in tumor samples was analyzed by immunohistochemistry.LincRNA-p21 was down-regulated in tumor tissue, but no association was observed with TP53 mutational status. High lincRNA-p21 levels were associated with poor CSS in all patients (p = 0.032). When patients were classified according to histological subtypes, the impact of lincRNA-p21 was confined to patients with adenocarcinoma in both time to relapse (p = 0.006) and CSS (p0.001). To explain the poor outcome of patients with high lincRNA-p21 expression, we studied the role of lincRNA-p21 in angiogenesis in vitro and observed a global downregulation in the expression of angiogenesis-related genes when lincRNA-p21 was inhibited. Moreover, supernatants from lincRNA-p21-inhibited cells were significantly less angiogenic and had lower levels of secreted vascular endothelial growth factor A than controls did. Finally, tumor samples with high lincRNA-p21 levels had higher microvascular density.Our findings suggest that lincRNA-p21 affects outcome in patients with NSCLC adenocarcinoma through the regulation of angiogenesis.

Background:Tofacitinib (TOFA) is the first JAKi approved for psoriatic arthritis (PsA) in Europe (July 2018). TOFA has shown efficacy in refractory patients to anti-TNF in Randomized Clinical Trials (RCT) (Gladman D. NEJM 2017; 377: 1525-36).Objectives:To assess efficacy and safety of TOFA in clinical practice (CP). To compare the profile of CP with RCTMethods:Study of 87 patients of CP with PsA treated with TOFA; Results are expressed as percentage, mean±SD or median [IRQ].Results:87 patients (28♀/59♂), mean age of 52.8±11.4 years (Table 1). Pattern of joint involvement was: peripheral (n=60), axial (1) and mixed (26). Presented also enthesitis (49.4%), nail involvement (30.2%) and dactylitis (31%).Prior TOFA, most patients (80%) received oral prednisone, synthetic immunosuppressants (mean 2.3±0.9) and biological therapy (BT) (3.6±1.9): etanercept (n=58), adalimumab (54), infliximab (31), golimumab (37), certolizumab (30), secukinumab (54), ustekinumab (39) and ixekizumab (2). Apremilast was used in 17.After a mean follow-up of 12.3±9.3 years from PsA diagnosis, TOFA was started (5 mg/12 h). In 48 (55.2%) TOFA was used in combined therapy: methotrexate (30) and leflunomide (15). In the remaining 39, monotherapy was prescribed.Patients of CP compared with RCT have a longer duration of PsA, worst functional disability (HAQ) and received a higher proportion of corticosteroids and BT (anti-TNF and non-anti-TNF) (Table 1).Patients improved in activity indexes (PASI, DAS28, DAPSA) and laboratory test (table 2). Minor side effects were reported in 21 patients (gastrointestinal symptoms), and TOFA was discontinued in 29 due to inefficiency mainly.Conclusion:Patients of CP had a longer evolution and received a greater number of biologics than those of RCT. TOFA as in RCT seems effective, rapid and relatively safe for refractory PsA.Table 1.Baseline featuresCLINICAL TRIALGladmanN=131CLINICAL PRACTICEN=87Age, years (mean±SD)49.5±12.352.8±11.4Sex, n (%)67M/64F (51/49)59M/28F (68/32.2)Duration PsA, years (mean±SD)9.6±7.612,3±9.3HAQ-DI1.3±0.71.4±0.7 (n=26)Swollen joint count, mean±SD12.1±10.65.7±5.8Painful joint count, mean±SD20.5±13.08.0±6.6Elevated CRP, n (%)85 (65)55 (63.2)PASI score, median [IQR]7.6 [0.6-32.2]9.0 [4.2-15]Oral glucocorticoid, n (%)37(28)44(50.5)Concomitant synthetic DMARDs, n (%)- Methotrexate98 (75)30 (34.4)- Leflunomide12 (9)15 (17.2)- Sulfasalazine21 (16)6 (6.9)- Others2 (2)N. of previous TNF inhibitors, mean±SD1.7±1.02.4±1.4Previous use of other biological no anti-TNF, n (%)11 (8)68 (78.2)Table 2.Table 2. Improvement at 1st, 6thand 12thmonthBaselinen=871st monthn=776th monthn=5212th monthn=20Nail involvement, n (%)17 (19.5)Improvement, n (%)5 (35.7)6 (60)5 (83.3)Enthesitis, n (%)28 (32.2)Improvement, n (%)8 (47.1)10 (58.8)3 (50)Dactylitis, n (%)16 (18.4)Improvement, n (%)9 (69.2)6 (85.7)0 (0)CRP mg/dl, median [IQR]1.9 [0.3-5]0.5 [0.1-2.2]0.5 [0.3-1.2]0.4 [0.4-3.7]p (vs baseline)0.0040.0050.66DAS28, median [IQR]4.8 [4.1-5.403.7 [2.8-4.6]2.8 [2.2-3.8]2.9 [2.2-3.7]p (vs baseline)DAPSA, median [IQR]28 [18.41-34.05]15.5 [10.1-25.7]9 [6.07-15]4.3 [2.4-8]p (vs baseline)PASI, median [IQR]5 [1-14]1.4 [0-7]0 [0-4]0.05 [0-2.7]p (vs baseline)0.1920.1050.300Disclosure of Interests:E. Galindez: None declared, D. Prieto-Peña: None declared, José Luis Martín-Varillas Grant/research support from: AbbVie, Pfizer, Janssen and Celgene, Speakers bureau: Pfizer and Lilly, Beatriz Joven-Ibáñez Speakers bureau: Abbvie, Celgene, Janssen, Merck Sharp & Dohme, Novartis, Pfizer, Olga Rusinovich: None declared, RAQUEL ALMODOVAR Speakers bureau: Abbvie, Celgene, Janssen, Lilly, Novartis, Pfizer., Juanjo J Alegre-Sancho Consultant of: UCB, Roche, Sanofi, Boehringer, Celltrion, Paid instructor for: GSK, Speakers bureau: MSD, GSK, Lilly, Sanofi, Roche, UCB, Actelion, Pfizer, Abbvie, Novartis, LARA MENDEZ DIAZ: None declared, Agusti Sellas-Fernández Speakers bureau: Abbott, Lilly, Celgene, Pfizer, Schering-Plough, Janssen, Novartis, and Nordic Pharma, À Martínez-Ferrer: None declared, Rosario Garcia de Vicuna Grant/research support from: BMS, Lilly, MSD, Novartis, Roche, Consultant of: Abbvie, Biogen, BMS, Celltrion, Gebro, Lilly, Mylan, Pfizer, Sandoz, Sanofi, Paid instructor for: Lilly, Speakers bureau: BMS, Lilly, Pfizer, Sandoz, Sanofi, Clara Ventín-Rodríguez: None declared, Julio Ramirez: None declared, Manuel Moreno: None declared, Maria jose Moreno: None declared, María del Carmen Castro Villegas: None declared, Antia Crespo Golmar: None declared, Natalia Palmou-Fontana: None declared, FRANCISCO ORTIZ SANJUAN: None declared, Ximena Elizabeth Larco Rojas: None declared, Antonio Juan Mas: None declared, Christian Y Soleto: None declared, Iñigo Gorostiza: None declared, Miguel A González-Gay Grant/research support from: Pfizer, Abbvie, MSD, Speakers bureau: Pfizer, Abbvie, MSD, Ricardo Blanco Grant/research support from: AbbVie, MSD, and Roche, Speakers bureau: AbbVie, Pfizer, Roche, Bristol-Myers, Janssen, and MSD

Background Pulmonary vascular abnormalities are a characteristic feature of chronic obstructive pulmonary disease (COPD). Cigarette smoking is the most important risk factor for COPD. It is believed that its constant exposure triggers endothelial cell damage and vascular remodelling. Under pathological conditions, progenitor cells (PCs) are mobilized from the bone marrow and recruited to sites of vascular injury. The aim of the study was to investigate whether in COPD the number of circulating PCs is related to the presence of bone marrow-derived cells in pulmonary arteries and the association of these phenomena to both systemic and pulmonary endothelial dysfunction. Methods Thirty-nine subjects, 25 with COPD, undergoing pulmonary resection because of a localized carcinoma, were included. The number of circulating PCs was assessed by flow cytometry using a triple combination of antibodies against CD45, CD133 and CD34. Infiltrating CD45+ cells were identified by immunohistochemistry in pulmonary arteries. Endothelial function in systemic and pulmonary arteries was measured by flow-mediated dilation and adenosine diphosphate-induced vasodilation, respectively. Results COPD patients had reduced numbers of circulating PCs (p

Objectives: Tumor associated fibroblasts (TAFs) are essential contributors of the progression of non-small cell lung cancer (NSCLC). Most lung TAFs exhibit an activated phenotype characterized by the expression of α-SMA and fibrillar collagens. However, the prognostic value of these activation markers in NSCLC remains unclear. Material and methods: We conducted a quantitative image analysis of α-SMA immunostaining and picrosirius red staining of fibrillar collagens imaged by bright-field and polarized microscopy, respectively, using tissue microarrays with samples from 220 surgical patients, which elicited a percentage of positive staining area for each marker and patient. Results: Kaplan-Meier curves showed that all TAF activation markers were significantly associated with poor survival, and their prognostic value was independent of TNM staging as revealed by multivariate analysis, which elicited an adjusted increased risk of death after 3 years of 129% and 94% for fibrillar collagens imaged with bright-field (p = 0.004) and polarized light (p = 0.003), respectively, and of 89% for α-SMA (p = 0.009). We also found a significant association between all TAF activation markers and tumor necrosis, which is often indicative of hypoxia, supporting a pathologic link between tumor desmoplasia and necrosis/hypoxia. Conclusions: Our findings identify patients with large histologic coverage of fibrillar collagens and α-SMA + TAFs to be at higher risk of recurrence and death, supporting that they could be considered for adjuvant therapy. This work was further supported bygrants from the Ministerio de Economía y Competitividad (MINECO/FEDER, UE) (PI13/02368 and SAF2016-79527-R to JA, FIS 12/02040to EM and FIS 12/02534 to EB), Fundació Privada Cellex (to JA),Generalitat de Catalunya AGAUR (SGR 661 to JA, SGR 801 to EM),Junta Provincial de Barcelona de l’Associació Espanyola Contra elCàncer (AECC B16-917 to JA), Fundació Catalana de Pneumologia (toEM), Sociedad Española de Neumología y Cirugía Torácica–SEPAR (PIIOncología Torácica to EM, < /GN6 > SEPAR 437 < /GN6 > to NR),and beca CONICYT (to N.D-V.)

Background c-Kit + lung stem cells have been described in the human healthy lung. Their potential relation with smoking and/or chronic obstructive pulmonary disease (COPD) is unknown. Methods We characterized and compared c-Kit+ cells in lung tissue of 12 never smokers (NS), 15 smokers with normal spirometry (S) and 44 COPD patients who required lung resectional surgery. Flow cytometry (FACS) was used to characterize c-Kit+ cells in fresh lung tissue disaggregates, and immunofluorescence (IF) for further characterization and to determine their location in OCT- embedded lung tissue. Results We identified 4 c-Kit+ cell populations, with similar proportions in NS, S and COPD: (1) By FACS, c-Kithigh/CD45+ cells (4.03 ± 2.97% (NS), 3.96 ± 5.30% (S), and 5.20 ± 3.44% (COPD)). By IF, these cells were tryptase+ (hence, mast cells) and located around the airways; (2) By IF, c-Kitlow/CD45+/triptase- (0.07 ± 0.06 (NS), 0.03 ± 0.02 (S), and 0.06 ± 0.07 (COPD) cells/field), which likely correspond to innate lymphoid cells; (3) By FACS, c-Kitlow/CD45-/CD34+ (0.95 ± 0.84% (NS), 1.14 ± 0.94% (S) and 0.95 ± 1.38% (COPD)). By IF these cells were c-Kitlow/CD45-/CD31+, suggesting an endothelial lineage, and were predominantly located in the alveolar wall; and, (4) by FACS, an infrequent c-Kitlow/CD45-/CD34- population (0.09 ± 0.14% (NS), 0.08 ± 0.09% (S) and 0.08 ± 0.11% (COPD)) compatible with a putative lung stem cell population. Yet, IF failed to detect them and we could not isolate or grow them, thus questioning the existence of c-Kit+ lung stem-cells. Conclusions The adult human lung contains a mixture of c-Kit+ cells, unlikely to be lung stem cells, which are independent of smoking status and/or presence of COPD. Electronic supplementary material The online version of this article (10.1186/s12890-018-0688-3) contains supplementary material, which is available to authorized users.

©2014 AACR. The crucial role of tumor-associated fibroblasts (TAF) in cancer progression is now clear in non-small cell lung cancer (NSCLC). However, therapies against TAFs are limited due to a lack of understanding in the subtype-specific mechanisms underlying their accumulation. Here, the mechanical (i.e., matrix rigidity) and soluble mitogenic cues that drive the accumulation of TAFs from major NSCLC subtypes: adenocarcinoma (ADC) and squamous cell carcinoma (SCC) were dissected. Fibroblasts were cultured on substrata engineered to exhibit normal- or tumor-like stiffnesses at different serumconcentrations, and critical regulatory processes were elucidated. In control fibroblasts from nonmalignant tissue, matrix stiffening alone increased fibroblast accumulation, and this mechanical effect was dominant or comparable with that of soluble growth factors up to 0.5% serum. The stimulatory cues ofmatrix rigidity were driven by β1 integrin mechanosensing through FAK (pY397), and were associated with a posttranscriptionally driven rise in β1 integrin expression. The latter mechano-regulatory circuit was also observed in TAFs but in a subtype-specific fashion, because SCC-TAFs exhibited higher FAK (pY397), β1 expression, and ERK1/2 (pT202/Y204) than ADC-TAFs. Moreover, matrix stiffening induced a larger TAF accumulation in SCC-TAFs (>50%) compared with ADC-TAFs (10%-20%). In contrast, SCC-TAFs were largely serum desensitized, whereas ADC-TAFs responded to high serum concentration only. These findings provide the first evidence of subtype-specific regulation of NSCLC-TAF accumulation. Furthermore, these data support that therapies aiming to restore normal lung elasticity and/or β1 integrin-dependent mechano regulation may be effective against SCC-TAFs, whereas inhibiting stromal growth factor signaling may be effective against ADC-TAFs. Implications: This study reveals distinct mechanisms underlying the abnormal accumulation of tumor-supporting fibroblasts in two major subtypes of lung cancer, which will assist the development of personalized therapies against these cells., Ministerio de Economía y Competitividad (SAF2009-13243 and PI13/02368, to J. Alcaraz; PS09/01377, to N. Reguart; SAF2012-34916, to F. Rodríguez-Pascual), AECC (10/103; to N. Reguart and J. Alcaraz), SEPAR (2009-853; to N. Reguart), and predoctoral fellowships from the Fundacio Cellex (to M. Puig),Ministerio de Educación (to A. Giménez), CONACYT (to R. Lugo), and COLCIENCIAS (to A. Velasquez).

In 2011, the Spanish Society of Medical Oncology and the Spanish Society of Pathology started a joint project to establish recommendations on biomarker testing in patients with advanced non-small-cell lung cancer based on the current evidence. Most of these recommendations are still valid, but new evidence requires some aspects to be updated. Specifically, the recommendation about which biomarkers to test in which patients is being amended, and the best way to manage tumour samples and minimum requirements for biomarker test material are defined. Suitable techniques for testing for epidermal growth factor receptor mutations and anaplastic lymphoma kinase rearrangement are also reviewed, and a consensus is reached on which situations warrant re-biopsy.

Background In resected non-small cell lung cancer (NSCLC) considerable recurrence rates has been reported (30%). Tumor secretes small extracellular vesicles known as exosomes, which plays a role in the metastasis process. Pulmonary tumor-draining vein blood has proved to be better source than peripheral vein to analyze the prognosis impact of exosomes (Navarro A, et al. Cancers 2019). LincRNA-p21 is a long non-coding RNA which upregulation in tumor tissue induced by hypoxia was related to poor prognosis in NSCLC (Castellano JJ. et al. JTO 2016). We aimed to evaluate the presence of lincRNA-p21 in exosomes and its potential as prognosis biomarker in resected NSCLC patients. Methods H23 and H1299 NSCLC cell lines were cultured in exosome-free medium 48h under normoxic/hypoxic conditions. LincRNA-p21 was inhibited using a siRNA and quantified using a TaqMan assay. Exosomes were purified by ultracentrifugation and characterized by electron microscopy, western blot and Nanosight. LincRNA-p21 was analyzed in exosomes from pulmonary tumor-draining vein, obtained during surgery, from 54 NSCLC patients. Results LincRNA-p21 was present in exosomes showing 18 times more representation than in their origin cells. Under hypoxic conditions exosomal lincRNA-p21 was upregulated in both cell lines (p Conclusions Tumor exosomes carry LincRNA-p21, which increases under hypoxia, and its analysis in tumor-draining vein arises as a promising prognosis marker. Legal entity responsible for the study The authors. Funding SAF2017-88606-P (AEI/FEDER, UE). Disclosure All authors have declared no conflicts of interest.

Background Little is known about the structure and function relationships of pulmonary vessels in the most severe chronic obstructive pulmonary disease (COPD) spectrum. We investigated morphometric, cellular, and physiologic characteristics of pulmonary arteries from COPD patients undergoing bilateral lung transplant. Methods Seventeen patients with very severe COPD (forced expiratory volume in 1 second, 24% ± 7%) were assessed using inert gas exchange and pulmonary hemodynamics while breathing ambient air and 100% oxygen. Morphometry, in vitro reactivity to hypoxia, and inflammatory cell counts of pulmonary arteries were measured in explanted lungs. Results Patients had moderate ventilation-perfusion imbalance along with mild release of hypoxic pulmonary vasoconstriction. Mild pulmonary hypertension was observed in 7 patients. Explanted lungs had predominant emphysema with mild small airway involvement. In vitro reactivity was modestly altered, with relatively preserved endothelium-dependent relaxation, and vascular remodelling was discrete, with intense CD8 + T lymphocytes infiltrate. In vitro reactivity correlated with pulmonary vascular resistance (on ambient air) and oxygen-induced pulmonary artery pressure changes. Patients with pulmonary hypertension had more severe morphologic and physiologic emphysema. Conclusions In end-stage COPD patients undergoing lung transplant, pulmonary vascular involvement is unexpectedly modest, with low-grade endothelial dysfunction. In this sub-set of COPD patients, pulmonary emphysema may constitute the major determinant of the presence of pulmonary hypertension.

NKX2-1, a key molecule in lung development, is strongly expressed in lung cancer, especially in adenocarcinoma patients. Recently, two microRNAs interacting with NKX2-1 have been described: miR-365 that down-regulate NKX2-1; and miR-33a that is directly upregulated. To date, in non-small cell lung cancer (NSCLC) contradictory prognostic information about NKX2-1 and its microRNAs has been described. Aim: Elucidate the prognostic role of NKX2-1 and its microRNAs in resected NSCLC patients. Methods: mRNA and microRNA expression was determined using TaqMan assay in 142 NSCLC patients. The cohort was molecularly characterized including mutational status of TP53, EGFR and KRAS. Results: Locally advanced stages (III) had higher NKX2-1 expression than early stages (I-II) (p=0.002). High NKX2-1 expression was associated with longer disease-free survival (DFS) only in early stages (p=0.0389). Regarding to TP53 status, NKX2-1 was significantly downregulated in TP53-mutated patients (p=0.0087). Interestingly, TP53-WT patients with high expression of NKX2-1 had longer overall survival (OS) (p=0.0274) and longer DFS (p=0.00276). A negative correlation between NKX2-1 and miR-365 was found (r=-0.342, p Conclusions: In our cohort NKX2-1 impacts prognostic in early stage NSCLC particularly in patients without TP53 mutation, but no prognostic value was confirmed for its associated microRNAs. Supported by PFR Hospital Clinic de Barcelona, Becario SEPAR 2015, Mutual Medica 2015.

Aim To evaluate the clinical relevance of the presence of lymphocytes in transbronchial needle aspiration (TBNA) samples from pathological mediastinal lymph nodes in patients with suspected lung cancer. Methods Retrospective observational study evaluating the negative predictive value (NPV) of TBNA samples with evidence of lymphocytes but no atypical cells. Results A total of 266 TBNA were performed in 252 patients with pathological mediastinal lymphadenopathy. One hundred and fifteen TBNA samples showed evidence of malignant cells (43%) and 94 (35%) samples were considered inadequate (absence of adequate cytological material or exclusive presence of bronchial epithelial cells). Out of the 57 TBNA samples remaining that contained lymphocytes without atypia (21%), 15 could not be confirmed. Thirty-two TBNA samples were confirmed with alternative diagnostic techniques and 10 were confirmed after clinical and radiological follow-up. The NPV of the 32 samples that were confirmed with alternative diagnostic techniques was 84%, which decreased to 76% when we included the 10 TBNA samples confirmed after clinical and radiological follow-up. Conclusions The presence of lymphocytes without atypia in TBNA samples does not rule out the neoplastic invasion of the lymph node analyzed.

The objectives of the study were to validate a model of methicillin-resistant Staphylococcus aureus (MRSA) pneumonia in ventilated piglets and to study the time-course of biological markers and histopathological changes. 12 piglets were intubated and inoculated with 15 mL of a suspension of 10 6 colony forming units of MRSA in every lobe through the bronchoscope channel. The piglets were ventilated for 12 h (n = 6) and 24 h (n = 6). Clinical parameters were assessed every 6 h and pro-inflammatory cytokines were measured in serum and in bronchoalveolar lavage (BAL) at baseline and sacrifice. Histopathology of each lobe and cultures from blood, lungs and BAL were performed. Animals developed histopathological evidence of pneumonia at necropsy. At 12 h, pneumonia was present in all animals and was severe pneumonia at 24 h. Microbiological studies confirmed the presence of MRSA. A significant increase in interleukin (IL)-6, IL-8 and tumour necrosis factor-α values was seen in BAL at 24 h and IL-6 at 12 h. In serum, only IL-6 levels had increased significantly at 24 h. In ventilated piglets, bronchoscopic inoculation of MRSA induces pneumonia at 12 h and severe pneumonia at 24 h. This severity was associated with a corresponding increase in systemic and local inflammatory response.

Potassium channels are important in pulmonary circulation because they have been closely related to hypoxic pulmonary vasoconstriction (HPV). The objective of the study was to determine whether structural changes in pulmonary arteries (PA), such as those observed in patients with chronic obstructive pulmonary disease (COPD), might be associated with changes in the mRNA expression of both BK(Ca) and K(V) channels and their potential relationship with HPV.PA (about 1.5 mm in diameter) were obtained from 16 patients who underwent resective lung surgery. Intimal thickening was evaluated morphometrically. mRNA expression of BK(Ca), K(V)1.2, K(V)1.5, K(V)2.1 and K(V)3.1 was evaluated by RT-PCR in PA homogenates. Endothelial function and HPV were assessed in vitro in isolated PA using an organ bath.Intimal enlargement was closely associated with an increase in the expression of BK(Ca) channel (r=0.57, p0.05). Pulmonary arteries incubated with charybdotoxin, a BK(Ca) channel blocking agent, showed lower response to endothelium-dependent vasodilators indicating its contribution to reduce vascular tone. Pulmonary arteries with more pronounced responses to hypoxia were those with greater gene expression of BK(Ca) channels, suggesting a potential role in attenuating HPV (r=0.52, p0.05). No changes in the expression of K(V) channels were found in remodeled arteries.Structural changes of PA in COPD could alter the response to hypoxia due to changes in BK(Ca) potassium channel distribution. Since BK(Ca) channels contribute to diminish vascular tone, their increased expression in remodeled PA might play a role in attenuating HPV.

Streptococcus pneumoniae is the most common causative pathogen of community-acquired pneumonia (CAP). Often, patients with pneumococcal CAP are admitted into intensive care units, particularly when affected by invasive serotypes.

Progenitor cells of bone marrow origin migrate to injured vessels, where they may contribute to endothelial maintenance and vessel remodeling through vascular endothelial growth factor (VEGF)-related signals. To what extent progenitor cells may play a role in vascular changes occurring in patients with chronic obstructive pulmonary disease (COPD) remains undetermined. In this study we sought to identify vascular progenitor cells in pulmonary arteries of patients with COPD and to investigate whether the presence of these cells could be related to changes in endothelial function or the expression of VEGF. Pulmonary arteries of nine patients with COPD and six control subjects were studied. Scanning electron microscopy demonstrated areas of denuded endothelium in the arteries of patients with COPD. Vascular progenitor cells were identified by immunohistochemistry and immunogold using antibodies against AC133, CD34, and CD45. AC133+ cells were localized in the endothelial surface, close to denuded areas. The number of AC133+ and CD45+ cells in pulmonary arteries was greater in patients with COPD than in control subjects. The number of AC133+ cells correlated with the response of pulmonary artery rings to hypoxic stimulus. AC133+ and CD45+ cells were also identified in the intimal layer. The wall thickness correlated with the number of progenitor cells in the intima and with VEGF and VEGF receptor-2 mRNA expression. We conclude that patients with COPD show an increased number of bone marrow-derived progenitor cells in pulmonary arteries. These cells seem to contribute to ongoing endothelial repair, but they might also be involved in the pathogenesis of pulmonary vascular remodeling.

Background: Sensitivity to glucocorticoids may be related to the concentration of glucocorticoid receptors α (GRα) and s (GRs). A study was undertaken to assess GRα and GRs expression in steroid insensitive interstitial lung disease (idiopathic pulmonary fibrosis (IPF)) and steroid sensitive interstitial lung diseases (sarcoidosis and cryptogenic organising pneumonia (COP)). Methods: Lung tissue was obtained from control subjects and from patients with IPF, sarcoidosis, and COP. Pulmonary function tests were carried out at the time of lung biopsy and every 3 months. GRα and GRs expression was evaluated by both competitive RT-PCR and immunohistochemistry. Data are presented as median and 25–75th percentile. Results: GRα mRNA expression (105 cDNA copies/µg total RNA) was higher in patients with steroid sensitive interstitial lung diseases (10.0; 7.8–14.9; n = 11) than in patients with IPF (4.4; 3.2–6.6; n = 19; p

Chronic obstructive pulmonary disease (COPD) is associated with structural and functional changes in the pulmonary circulation that commence at an early stage. To investigate whether vascular endothelial growth factor (VEGF) might be implicated as a mediator in COPD-associated pulmonary vascular changes, we studied surgical specimens obtained from 19 nonsmokers, 21 smokers with normal lung function, 28 patients with moderate COPD, and 10 patients with severe emphysema. The expression of VEGF in pulmonary muscular arteries was evaluated by immunohistochemistry, its protein content in lung tissue by Western blot analysis, and VEGF mRNA and its isoforms were analyzed by reverse transcription-polymerase chain reaction. The immunohistochemical expression of VEGF was increased in pulmonary arteries of smokers (median, 68% [interquartile range, 60-88]) and patients with moderate COPD (77% [63-82]), compared with nonsmokers (53% [40-63]) (p0.05 each). The expression of VEGF in smooth muscle cells correlated with the thickness of the vessel wall (r = 0.38, p0.01). VEGF protein content in lung tissue was reduced in severe emphysema, where reverse transcription-polymerase chain reaction demonstrated a lower proportion of the VEGF189 isoform. In conclusion, the expression of VEGF varies according to the severity of COPD and might be involved in the pathogenesis of pulmonary vascular remodeling at early stages of the disease.

// Alfons Navarro 1 , Rut Tejero 1 , Nuria Vinolas 2 , Anna Cordeiro 1 , Ramon M. Marrades 3 , Dolors Fuster 1 , Oriol Caritg 1 , Jorge Moises 3 , Carmen Munoz 1 , Laureano Molins 5 , Josep Ramirez 4 , Mariano Monzo 1 1 Molecular Oncology and Embryology Laboratory, Human Anatomy Unit, School of Medicine, University of Barcelona, IDIBAPS, Barcelona, Spain 2 Department of Medical Oncology, Institut Clinic Malalties Hemato-Oncologiques (ICMHO), Hospital Clinic de Barcelona, University of Barcelona, IDIBAPS, Barcelona, Spain 3 Department of Pneumology, Institut Clinic del Torax (ICT), Hospital Clinic de Barcelona, University of Barcelona, IDIBAPS, CIBER de Enfermedades Respiratorias (CIBERES), Barcelona, Spain 4 Department of Pathology, Centro de Diagnostico Biomedico (CDB), Hospital Clinic de Barcelona, University of Barcelona, IDIBAPS, CIBERES, Barcelona, Spain 5 Department of Thoracic Surgery, Institut Clinic del Torax (ICT), Hospital Clinic de Barcelona, University of Barcelona, Barcelona, Spain Correspondence to: Alfons Navarro, e-mail: anavarroponz@ub.edu Keywords: PIWI proteins, piwiRNAs, PIWIL1, PIWIL4, NSCLC Received: November 14, 2014 Accepted: December 21, 2014 Published: January 23, 2015 ABSTRACT The expression of Piwi-interacting RNAs, small RNAs that bind to PIWI proteins, was until recently believed to be limited to germinal stem cells. We have studied the expression of PIWI genes during human lung embryogenesis and in paired tumor and normal tissue prospectively collected from 71 resected non-small-cell lung cancer patients. The mRNA expression analysis showed that PIWIL1 was highly expressed in 7-week embryos and downregulated during the subsequent weeks of development. PIWIL1 was expressed in 11 of the tumor samples but in none of the normal tissue samples. These results were validated by immunohistochemistry, showing faint cytoplasmic reactivity in the PIWIL1 -positive samples. Interestingly, the patients expressing PIWIL1 had a shorter time to relapse (TTR) ( p = 0.006) and overall survival (OS) ( p = 0.0076) than those without PIWIL1 expression. PIWIL2 and 4 were downregulated in tumor tissue in comparison to the normal tissue ( p < 0.001) and the patients with lower levels of PIWIL4 had shorter TTR ( p = 0.048) and OS ( p = 0.033). In the multivariate analysis, PIWIL1 expression emerged as an independent prognostic marker. Using 5-Aza-dC treatment and bisulfite sequencing, we observed that PIWIL1 expression could be regulated in part by methylation. Finally, an in silico study identified a stem-cell expression signature associated with PIWIL1 expression.

Intimal enlargement of pulmonary arteries is an early change in chronic obstructive pulmonary disease (COPD). The cellular and extracellular components that are involved in this enlargement are unknown. The present study was designed to characterize the structural changes occurring in pulmonary muscular arteries in the initial disease stages. Lung specimens from patients with moderate COPD (n=8; forced expiratory volume in one second (FEV1), 66 +/- 10% predicted) and smokers without airflow obstruction (n=7; FEV1, 86 +/- 6% pred), were investigated by histochemistry to characterize extracellular matrix proteins and by immunohistochemistry to identify intrinsic cells of the vascular wall. In both COPD patients and smokers, the majority of cells present in the enlarged intimas were stained by specific smooth muscle cell (SMC) markers. No staining with endothelial or fibroblast markers was shown. A proportion of SMCs did not stain with desmin, suggesting cellular heterogeneity in this population. Elastin was the most abundant extracellular matrix protein and collagen was seen in a lower proportion. The amount of collagen was related to the intimal thickness (p

To the Editors: Obstructive sleep apnoea (OSA) is a very prevalent syndrome that induces or aggravates cardiovascular, metabolic and neurocognitive disorders. Among the various challenges imposed on a patient by OSA as a consequence of apnoeas (sleep disruption, increased inspiratory efforts and recurrent hypoxaemia), intermittent hypoxia plays a major role in the pathophysiology of this sleep breathing disorder. It has been well established that hypoxia plays an important role in regulating the various stages of tumour formation and progression [1]. Accordingly, the aim of the experimental study reported here was to test the hypothesis that high-rate intermittent hypoxia with a time course similar to that found in OSA (one or more hypoxic event per minute) enhances tumour growth. To avoid the interaction of any comorbidity, this investigation was carried out in a well-controlled animal model where the main variable under study was intermittent hypoxia. This study, which was approved by the Ethical Committee for Animal Research of the University of Barcelona (Barcelona, Spain), was conducted in 15 pathogen-free male C57BL/6J mice (25–30 g) using a conventional murine melanoma model consisting of tumour induction by subcutaneous injection of 106 B16F10 melanoma cells (ATCC-CRL-6475; American Type Culture Collection, Manassas, VA, USA) in the left flank region of the mouse [2]. This is a widely used cancer model in experimental research [3, 4], which has a response to hypoxia that is representative of a variety of cancer types …

BACKGROUND A study was undertaken to assess the diagnostic value of different clinical criteria and the impact of microbiological testing on the accuracy of clinical diagnosis of suspected ventilator associated pneumonia (VAP). METHODS Twenty five deceased mechanically ventilated patients were studied prospectively. Immediately after death, multiple bilateral lung biopsy specimens (16 specimens/patient) were obtained for histological examination and quantitative lung cultures. The presence of both histological pneumonia and positive lung cultures was used as a reference test. RESULTS The presence of infiltrates on the chest radiograph and two of three clinical criteria (leucocytosis, purulent secretions, fever) had a sensitivity of 69% and a specificity of 75%; the corresponding numbers for the clinical pulmonary infection score (CPIS) were 77% and 42%. Non-invasive as well as invasive sampling techniques had comparable values. The combination of all techniques achieved a sensitivity of 85% and a specificity of 50%, and these values remained virtually unchanged despite the presence of previous treatment with antibiotics. When microbiological results were added to clinical criteria, adequate diagnoses originating from microbiological results which might have corrected false positive and false negative clinical judgements (n = 5) were countered by a similar proportion of inadequate diagnoses (n = 6). CONCLUSIONS Clinical criteria had reasonable diagnostic values. CPIS was not superior to conventional clinical criteria. Non-invasive and invasive sampling techniques had diagnostic values comparable to clinical criteria. An algorithm guiding antibiotic treatment exclusively by microbiological results does not increase the overall diagnostic accuracy and carries the risk of undertreatment.

Fundacion Mutua Madrilena; Fondo de Investigaciones Sanitarias [PI11/02866], Conde, E., Suárez-Gauthier, A., Benito, A., Garrido, P., García-Campelo, R., Biscuola, M., Paz-Ares, L., Hardisson, D., De Castro, J., Camacho, M.C., Rodriguez-Abreu, D., Abdulkader, I., Ramirez, J., Reguart, N., Salido, M., Pijuán, L., Arriola, E., Sanz, J., Folgueras, V., Villanueva, N., Gómez-Román, J., Hidalgo, M., López-Ríos, F.

Permanent cell cycle arrest through senescence has been previously regarded as a protective mechanism against tumor progression. In contrast, there is evidence that senescence in tumor-associated fibroblasts (TAFs) enhances tumor growth. Senescence has been reported in tumor associated fibroblasts (TAFs) from a growing list of selected cancer types. However, the presence of senescent TAFs in lung cancer remains undefined. To address this gap of knowledge, we examined common markers of senescence in primary TAFs from the 3 major non-small cell lung cancer (NSCLC) subtypes: adenocarcinoma (ADC), squamous cell carcinoma (SCC) and large cell carcinoma (LCC). Given the difficulties in gathering LCC-TAFs owing to the lower prevalence of LCC compared to the other subtypes, primary fibroblasts from 2 independent cell collections were used. We found an enrichment of the myofibroblast-like phenotype in TAFs regardless their histologic subtype, yet senescence was observed in LCC-TAFs only. Likewise, co-culturing normal lung fibroblasts with LCC (but not ADC or SCC) cancer cells was sufficient to induce senescence, and this induction was prevented in the presence of an antioxidant, indicating that it is mediated through oxidative stress. Of note, senescent fibroblasts provided growth and invasive advantages to LCC cells in culture and in vivo beyond those provided by control (non-senescent) fibroblasts. These results expand recent evidence that challenges the common assumption that lung TAFs are a heterogeneous myofibroblast-like cell population regardless their histologic subtype. Of note, because LCC often distinguishes itself in the clinic by its aggressive nature, our findings support that senescent or senescent-like TAFs may contribute to the selective aggressive behavior of LCC tumors. Citation Format: Roberto Lugo, Marta Gabasa, Francesca Andriani, Marta Puig, Federica Facchinetti, Josep Ramírez, Abel Gómez-Caro, Ugo Pastorino, Pere Gascón, Albert Davalos, Noemí Reguart, Luca Roz, Jordi Alcaraz. Cancer cell-stromal cell crosstalk drives fibroblast senescence and tumor progression in large cell carcinoma of the lung in culture and in vivo. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4103.

Cigarette smoke (CS) induces an inflammatory process in the lung that may underlie the development of chronic obstructive pulmonary disease (COPD). The nature and characteristics of this process have not been fully established in animal models. We aimed to evaluate the pulmonary inflammatory reaction and its involvement in structural changes in guinea pigs chronically exposed to CS. 19 Hartley guinea pigs were exposed to 7 cigarettes/day, during 3 or 6 months. 18 control guinea pigs were sham-exposed. Numbers of neutrophils, macrophages and eosinophils and lymphoid follicles were assessed in different lung structures. Airway and vessel morphometry, alveolar space size and collagen deposition were also quantified. After 6 months of exposure, CS-exposed guinea pigs showed increased numbers of neutrophils, macrophages and eosinophils in the airways, intrapulmonary vessels and alveolar septa, as well as lymphoid follicles. Increased numbers of muscularized intrapulmonary vessels were apparent at 3 months. After 6 months of exposure, the airway wall thickened and the alveolar space size increased. Collagen deposition was also apparent in airway walls and alveolar septa after 6 months' exposure. The magnitude of airway wall-thickening correlated with the number of infiltrating inflammatory cells, and the extension of collagen deposition correlated with alveolar space size. We conclude that in the guinea pig, 6 months of CS exposure induces inflammatory cell infiltrate in lung structures, at an intensity that correlates with airway remodelling. These changes resemble those observed in COPD, thus endorsing the pathogenic role of CS and the usefulness of this animal model for its study.

To assess the false-negative (FN) rate of positron emission tomography (PET)-chest computed tomography (CT) scan in clinical non-central cIA and cIB non-small-cell lung cancer (NSCLC) for mediastinal staging.Between January 2007 and December 2010, 402 patients with potentially operable NSCLC were assessed by thoracic CT scan and 18-fluoro-2-deoxy-d-glucose PET-CT for mediastinal staging and to detect extrathoracic metastases, of which 153 surgically treated patients (79 cIA and 74 cIB cases) were prospectively included in the study. Central tumours were excluded on the basis of CT scan criteria, defined as contact with the intrapulmonary main bronchi, pulmonary artery, pulmonary veins or the origin of the first segmental branches. CT scan was considered negative if lymph nodes were1 cm at the smaller diameter. 18FDG PET-CT was considered negative when the high maximum standard uptake value (SUVmax) was2.5. Non-invasive surgical staging was carried out in this group, and curative resection plus systematic mediastinal dissection was performed except in the event of unexpected oncological contraindication.Composite non-invasive staging (CT scan, PET-CT) showed a negative predictive value (NPV) of 92% (CI 83.6-96.8) in the cIA group and 85% (CI 74-92) in the cIB group. There were 6 of 79 (7.6%) false-negatives (FNs) in cIA and 11 of 74 (14.8%) in cIB. Multilevel pN2 were detected in four cases, all of them in the cIB group. The most frequently involved N2 was subcarinal (two cases) in cIA and right lower paratracheal (R4) and seven (five cases) in cIB. Occult (pN2) lymph nodes were more frequent in tumour sizes≥5 cm (pT2b, nine cases, four FNs, P=0.03), pN1, adenocarcinoma [excluding minimally invasive adenocarcinoma (MIA) and lepidic predominant growth (LPA)] (P=0.029) and female patients, but no other risk factors for mediastinal metastases were identified (age, clinical stage, tumour location, central or peripheral, P0.05). Multilevel pN2 was significantly more frequent in the cIB group (P0.03). In pT≤1 cm (T1a), NPV was significantly better (NPV=100%, P0.05) than the other subgroups studied (IA1 cm and IB).Composite results for non-invasive mediastinal staging (CT scan, PET-CT) showed 11% of FNs in cI stage (7.6% in non-central cIA and 14.8% in cIB). In tumours≤1 cm, NPV makes surgical staging unnecessary. In women with adenocarcinoma and non-central cIB, however, the high FN rate makes invasive staging necessary, particularly in pT2b to decrease the incidence of unexpected pN2 in thoracotomy.

In this report, we describe a patient who developed acute renal failure and fatal pulmonary hemorrhage. Autopsy examination revealed pauci-immune necrotizing glomerulonephritis and pauci-immune hemorrhagic alveolar capillaritis. No evidence of granulomatous inflammation or vasculitis other than capillaritis was found in either lungs or kidneys. In addition, histopathologic examination of the upper respiratory tract, liver, spleen, digestive tract, and skeletal muscle was completely normal. The patient's serum sample disclosed antiproteinase 3 antibodies with an immunofluorescence titer of 1:1,600. Although there have been reports of patients with pulmonary-renal syndrome in association with antiproteinase 3 antibodies in the absence of a well-known vasculitic disease, we have not so far seen a case documented by a detailed autopsy study such as is presented here. In view of our findings and those of some other reports on this clinicopathologic condition, we suggest that this might represent a distinct form of vasculitis with exclusive capillary involvement.

Background Obstructive sleep apnea (OSA) is associated with cardiovascular disorders, but the different comorbidities in OSA patients make it difficult to know their specific effects on the development of cardiovascular injury. The aim of the present study was to investigate whether recurrent obstructive apneas could lead to myocardial injury. Methods Thirty-six male Sprague–Dawley rats (300–350 g) were either acutely (3 h) or sustainably (5 h/day, for 10 days) subjected to obstructive apneas with a pattern of 15 s each, 60 apneas/h. Corresponding control groups were formed for the acute and sustained models. To assess the induction of systemic inflammation, IL1-β was measured in plasma. Ventricular tissue injury was evaluated by histological techniques (presence of inflammatory cell infiltration, eosin autofluorescence, and detection of apoptosis). Results After 3 h of obstructive apneas, a significant increase in IL1-β (64.9 ± 29.6 ng/μl) were observed with respect to the controls (7.3 ± 1.0 ng/μl), but no myocardial injury was present. Conversely to the acute model, the systemic inflammation triggered by obstructive apneas for 10 days was reduced. However, the percentage of area with enhanced eosin autofluorescence and of apoptotic cells (1.83 ± 0.35% and 24.4 ± 1.5%, respectively) was increased when compared to the control group (0.72 ± 0.20% and 5.0 ± 2.8%, respectively). Conclusions This study suggests that obstructive apneas are a potential source of early systemic and ventricular inflammation and myocardial cell injury after sustained apneas application, which could represent an initial phase in the progression of heart disease associated with OSA.

The upper airway of obstructive sleep apnoea patients is subjected to recurrent negative pressure swings promoting its collapse and reopening. The aim of the present study was to ascertain whether this mechanical stress induces upper airway inflammation in a rat model. The upper airway of Sprague-Dawley rats was subjected to a periodic pattern of recurrent negative (-40 cmH2O, 1 s) and positive (4 cmH2O, 2 s) pressures inducing collapse and reopening for 5 h. Rats that were instrumented but not subjected to negative pressure swings were used as controls. The gene expression of the pro-inflammatory biomarkers macrophage inflammatory protein (MIP)-2, tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta and P-selectin in the soft palate and larynx tissues was assessed by real-time PCR. A marked overexpression of MIP-2, TNF-alpha, IL-1beta and P-selectin (approximately 40-, 24-, 47- and 7-fold greater than controls, respectively) was observed in the larynx tissue; similar results were found in the soft palate tissue (approximately 14-, 7-, 35- and 11-fold greater than controls, respectively). Recurrent upper airway collapse and reopening mimicking those experienced by obstructive sleep apnoea patients triggered an early local inflammatory process. These results could explain the inflammation observed in the upper airway of obstructive sleep apnoea patients.

Extensive tracheal stenotic lesions caused by tracheomalacia or neoplasms represent a surgical challenge. Segmental tracheal substitution is sometimes required to obtain radical cure. We present an experimental study of 27 dogs undergoing replacement of the cervical trachea using a vascularized small bowel segment as a tubular graft. A silicone stent was placed in the lumen of the intestinal fragment and was removed the second week after operation. Endoscopic and histological examinations were performed between the first week and second month after operation, and rigidity of the graft was assessed in all cases. No evidence of anastomotic stricture or mucous formation was found. Microscopic examination showed the substitution of bowel mucosa by squamous epithelium as well as the development of connective tissue favoring the fixation of the skeletal muscular structures of the neck to the serous layer of the graft, thus avoiding collapse of the new airway.

Objectives The aim of this study was to evaluate the temporal association between the diagnosis of neoplasia and sarcoidosis in a large cohort of Spanish sarcoidosis patients. Methods We included 170 patients consecutively diagnosed with sarcoidosis. The diagnosis of sarcoidosis was based on clinical criteria and imaging studies, together with the histopathological demonstration of non-necrotizing granulomas, and excluding other granulomatous diseases. The diagnosis of neoplasia was recorded before and after the diagnosis of sarcoidosis. Hematological neoplasias were classified according to the 2008 WHO Classification. Results There were 111 (65%) women and 59 (35%) men (mean age at diagnosis, 47 years); 23 (13.5%) patients developed 25 neoplasias (two patients developed 2 neoplasias, before and after the diagnosis of sarcoidosis). There were 13 (52%) solid neoplasias, including pulmonary (n=3), thyroid (n=2), breast (n=2), melanoma (n=2) and cervix, testicular, renal and gastric cancer (1 case each). The remaining 12 (48%) were hematological neoplasias, mainly mature B-cell neoplasms (3 chronic lymphocytic leukemia, 3 extranodal MALT lymphomas and 2 diffuse large B-cell lymphoma), but also 2 acute leukemias (1 acute promyelocytic PML-PARA, 1 acute myeloid leukemia), 1 mature T-cell neoplasia (mycosis fungoide) and 1 myeloid neoplasia (essential thrombocytopenia). No significant differences were found in the main epidemiological, clinical or radiological features of sarcoidosis in patients with or without associated neoplasia, except for a statistical trend to a higher frequency of extrathoracic sarcoidosis in patients with associated cancer (70% vs 50%, p=0.11). Of the 23 patients with cancer, 5 were diagnosed before sarcoidosis and 16 after the diagnosis of sarcoidosis (10 solid and 6 hematological neoplasia); the 2 remaining patients developed a second neoplasia after the diagnosis of sarcoidosis. Conclusions We identified four states of association between sarcoidosis and malignancy: i) patients with sarcoidosis who develop solid tumors (43%); ii) development of hematologic malignancies, mainly sarcoidosis-lymphoma syndrome (26%); iii) development of sarcoidosis in patients with neoplasia, including paraneoplastic sarcoidosis (cancer discovered concurrently ±1 year of the diagnosis of sarcoidosis) in 3 patients (22%); iv) patients with sarcoidosis and previous cancer who developed a second and different neoplasia (9%). Disclosure of Interest None declared

Objectives To evaluate extrapulmonary activity and correlate baseline features at diagnosis with survival in a large cohort of patients with sarcoidosis. Methods We studied 170 patients consecutively diagnosed with sarcoidosis in the last 10 years. The diagnosis of sarcoidosis was based on clinical criteria and imaging studies, together with the histopathological demonstration of non-necrotizing granulomas, and the exclusion of other granulomatous diseases, especially infections (tuberculosis). Extrapulmonary involvement was defined according to the criteria defined by Judson et al, 2011. Results There were 111 (65%) women and 59 (35%) men, with a mean age at diagnosis of 47.49 years (range 17-92). The main clinical manifestations present at diagnosis were respiratory symptoms in 112 (66%) patients, erythema nodosum in 35 (21%), articular involvement in 35 (21%) and fever in 34 (20%) patients; incidental imaging findings in asymptomatic patients was found in 12% of cases. After a mean follow-up of 107.4 months, 21 (12%) patients died. The main causes of death were infections in 4 patients, cardiovascular disease in 2, pulmonary sarcoidosis in 1, neoplasia in 1 and other causes in 2 patients; in the remaining 11 patients, the cause of death was unknown or unclassifiable. Survivors had a lower mean age (44.82 yrs. vs. 65.95 yrs. p Conclusions Extrapulmonary involvement in sarcoidosis is not associated with mortality. An older age at diagnosis was the principal risk factor associated with death, while patients born outside Spain had a lower mortality rate, and mean ACE levels at diagnosis were inversely associated with survival. The very-low rate of deaths caused directly by sarcoidosis might explain our results. Disclosure of Interest None declared

Patients with chronic obstructive pulmonary disease (COPD) show impaired hypoxic pulmonary vasoconstriction that might contribute to abnormal gas exchange and could be related to endothelial dysfunction in pulmonary arteries. The aim of the study was to investigate the response of PA to hypoxic stimulus in vitro in COPD, and the role of endothelium-derived nitric oxide (NO) in this response. The pulmonary arteries of 25 patients who underwent lung resection were studied. Patients were divided into controls, COPD+normoxaemia (COPDN) and COPD+ hypoxaemia (COPDH). Hypoxic vasoconstriction (HV) was evaluated before and after stimulation or inhibition of the endothelial release of NO, and in the presence of exogenous NO. Compared with the other groups, HV was reduced in COPDH. The magnitude of HV correlated with the oxygen tension in arterial blood. The hypoxic stimulus induced greater contraction after stimulating endothelial release of NO, whereas its inhibition practically abolished HV. Exogenous NO completely inhibited HV. Maximal relaxation induced by endothelium-dependent vasodilators correlated with the magnitude of HV. In conclusion, pulmonary arteries of patients with chronic obstructive pulmonary disease and hypoxaemia have an impaired response to hypoxic stimulus, and the endothelial release of nitric oxide modulates hypoxic vasoconstriction. The depressed response of pulmonary arteries to hypoxia may contribute to abnormal gas exchange in chronic obstructive pulmonary disease.

The mechanisms underlying the abundance of tumor associated fibrobasts (TAFs) in non-small cell lung cancer (NSCLC) remain poorly understood. To address this gap of knowedge, we examined the relative contribution of mechanical (i.e. matrix rigidity) and soluble biochemical (i.e. serum) cues in the accumulation of primary TAFs from either adenocarcinoma (ADC) or squamous cell carcinoma (SQC) NSCLC patients. For this purpose, fibroblasts were cultured on collagen-I coated gels engineered to exhibit either normal- or tumor-like stiffnesses at different serum concentrations. In fibroblasts from non-malignant tissue, matrix stiffening alone was sufficient to increase fibroblast accumulation by 20%. In addition, matrix stiffening-induced accumulation was dominant or comparable to that due to soluble growth factors up to moderate serum concentrations. These mechanical stimulatory effects were β1 integrin-dependent, and were associated with both increased integrin mechanosensing through FAKpY397and a post-transcriptionally-driven rise in β1 integrin expression. The latter mechanoregulatory circuit could be extended to TAFs in a subtype-dependent fashion, since SQC-TAFs exhibited higher FAKpY397 and β1 expression than ADC-TAFs. Remarkably, SQC-TAF density was enhanced > 50% in response to matrix stiffening, whereas they were largely desensitized to serum. Conversely, substantial accumulation of ADC-TAFs required large serum concentration, whereas it was poorly induced by either matrix stiffening or low serum concentration. These results reveal that a therapeutic strategy aiming to inhibit soluble growth factors may be effective against ADC-TAFs, whereas a strategy aiming to rescue normal lung elasticity may be more efficient against SQC-TAFs. Citation Format: Jordi Alcaraz, Marta Puig, Roberto Lugo, Alícia Giménez, Adriana Velásquez, Roland Galgoczy, Josep Ramírez, Abel Gómez-Caro, Pere Gascón, Noemi Reguart. Matrix stiffening and β1 integrin promote fibroblast accumulation in lung squamous cell carcinomas but not in adenocarcinomas. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1089. doi:10.1158/1538-7445.AM2014-1089

Aim: In the SLCG large-scale screening, 16.6% of EGFR mutations were found, median PFS and OS for pts receiving erlotinib was 14 mos and 27 mos, respectively. The HR for the duration of PFS was 1.68 for the presence of L858R mutation in paired serum DNA (P = 0.02) (NEJM 2009; 361:958-67). Mutant EGFR NSCLC selectively activates STAT3 signaling which promotes cell survival and single EGFR inhibition can even enhance STAT3 activation. Growing evidence shows that targeting STAT3 or upstream or downstream components could improve the outcome. Hence, the resistance mechanisms should be revised, T790M is detected in 60% before therapy (reported by SLCG) and MET signaling can cause inter-receptor crosstalk with AXL, EPHA2, JAK1 and CDCP1. BIM mRNA expression is an independent predictive marker of response to erlotinib (reported by SLCG). High BIM can predict response and early adaptive resistance via IL6-STAT3-Bcl2. In those patients with low BIM, STAT3 can be activated through over-expression of AXL and EPHA2. Crosstalk between TGFb and NOTCH signaling can also be involved. Methods: Transcripts involving STAT3 signaling were examined from 80 EGFR mutant NSCLC pts, including BIM, STAT3 and SHP2. Also MET, AXL, Mer, EphA2 and Gas6 were analyzed. Together with TGFb and NOTCH constituents: ADAM17, NUMB, HES1, DUSP1, RBJ, LncRNA-ATB, SLUG, GLI and PKCi. Repurposing drugs, in combination with gefitinib, erlotinib and afatinib, were examined in cell cultures and in subcutaneous and orthotopic xenograft mice models. Results: Data will be presented on the correlation of BIM and SHP2 with regard to the mechanism of activation of STAT3 and the influence in response, PFS and OS in the 80 erlotinib-treated EGFR mutant NSCLC pts, and the influence of the above mentioned signaling constituents. Early data shows synergism of gefitinib with AXL inhibitor in the PC9-R cells. Also, activity of niclosamide and paclitaxel. Conclusions: Based on the results, the SLCG-GFCP will carry out a trial with erlotinib in combination with the most active repurposing drugs found, customized by BIM expression. Disclosure: All authors have declared no conflicts of interest.

25 consecutive inoperable or extended non-small cell lung cancer (NSCLC) patients (19 non-chemotherapy pretreated, 6 non-heavily pretreated) were given oral etoposide, 50 mg/m2/day for 21 successive days, every 4 weeks. 5 partial responses (PR), 9 disease stabilizations were achieved; the overall response rate of 20% (95% confidence interval, 4% to 36%) or 26% in non-pretreated patients. Median survival and PR duration probabilities were 6.7 months and 6.3 months, respectively. Alopecia excepted (96% of patients), non-haematological toxicity was mild. Haematological toxicity WHO grade II+III mainly consisted of leukopenia (28%).

Objective: To validate sampling techniques (tracheobronchial aspirates, protected specimen brush, and bronchoalveolar lavage, both conventional and protected) for the detection of ventilator-associated pneumonia (VAP) and causative microorganisms according to different histologic and microbiological references. Design: Immediate, multiple bilateral lung biopsy, postmortem study. Setting: Respiratory intensive care unit of a 1,000-bed teaching hospital. Patients: Twenty-five mechanically ventilated patients (>72 hrs) who died in our intensive care unit. Measurements: Lung tissue histologic examination and quantitative cultures (16 specimens/patient). The following four references for the diagnostic techniques were used: histology of guided lung biopsy, histology of blind lung biopsy, combined guided and blind lung biopsy histology and microbiology of lung tissue, and microbiology of lung tissue. Results: Sensitivities when histologic reference tests were used ranged from 16% to almost 40%, whereas specificity rates were always

Mediastinal metastasis of ovarian tumors are not rare as autopsy findings. Ovarian carcinomas usually spread by transcaelomic, lymphatic or haematogenous dissemination to peritoneum, pelvic and para-aortic lymph nodes, lung and pleura. A case of mediastinal metastasis of ovarian carcinoma is reported. A retrosternal mass was identified by CT scan and resected by VTC surgery.

Endothelial dysfunction and intimal thickening have been shown in pulmonary arteries (PA) of patients with mild chronic obstructive pulmonary disease (COPD). To investigate whether an inflammatory process related to tobacco smoking might be involved in the development of pulmonary vascular abnormalities in COPD, we characterized the inflammatory cell infiltrate and the endothelium-dependent relaxation in PA of 39 patients who underwent lung resection, divided into three groups: "nonsmokers" (n = 7); "smokers," with normal lung function (n = 12); and "COPD" (n = 20). Endothelium-dependent relaxation was assessed in vitro by exposing PA rings to adenosine diphosphate (ADP). Inflammatory cell types were identified by immunohistochemistry. PA of COPD patients developed lower relaxation in response to ADP than nonsmokers and smokers. The number of inflammatory cells was increased in PA of COPD compared with the other two groups. This cell infiltrate was largely constituted by T lymphocytes. The CD8(+) T-cell subset was increased in both smokers and COPD compared with nonsmokers, yielding a reduction of the CD4(+)/CD8(+) ratio. The intensity of the inflammatory infiltrate correlated with both the endothelium-dependent relaxation and the intimal thickness. We conclude that cigarette smoking induces a CD8(+) T-lymphocyte infiltrate in PA, which is associated with the impairment of the vessel's structure and function, suggesting the potential involvement of an inflammatory process in the pathogenesis of pulmonary vascular abnormalities in the early stage of COPD.

Langerhans’ cell histiocytosis (LCH) is a proliferative histiocytic disorder of unknown origin. The lesions of LCH contain single or multifocal proliferation of histiocytes similar in phenotype to dendritic Langerhans’ cells (1). The disorder ranges in clinical severity from a solitary eosinophilic granuloma of bone to a generalised disease with multiple organ involvement. Primary involvement of the lung is rare and only few isolated cases have been reported (2). Occasionally, pulmonary LCH has been reported to be associated with Hodgkin’s disease (3); the underlying pathogenic mechanism of such association remains unknown. Here, we report the case of a 19-year-old patient who developed pulmonary LCH 2 months after receiving an autologous haemopoietic progenitor cell transplantation (AHPCT) for Hodgkin’s disease.

The diagnosis of pulmonary candidiasis is still controversial. We undertook a prospective study on 25 non-neutropenic, mechanically ventilated (> 72 h) patients who died in our ICU with the aim of assessing the incidence and significance of the isolation of Candida species from quantitative cultures of immediate postmortem lung biopsies and different respiratory sampling techniques. Immediate postmortem respiratory samples (endotracheal aspirate, protected specimen brush [PSB], bronchoalveolar lavage [BAL], blind biopsies [average 14/patient], and bilateral bronchoscopically guided biopsies [two per patient]) were taken from all patients. Lung tissue specimens were histologically examined. Respiratory samples were classified as having Candida or otherwise. Ten (40%) patients had at least one pulmonary biopsy yielding Candida spp. Among these 10 patients with Candida isolates, only two had definite pulmonary candidiasis. A total of 470 microorganisms were isolated from 280 of 375 (77%) lung biopsy samples in all 25 patients. Candida species represented 9% (n = 40) of the isolates, corresponding to 10 patients (40%). In the 10 patients in whom Candida species was isolated from pulmonary biopsies, this was always associated with the isolation of the same microorganism from one of the sampling methods. Quantitative cultures of Candida species from different sampling methods correlated well among each other but could not discriminate the presence from absence of Candida pneumonia. A logistic regression model adjusted for the presence of antibiotics, days of antibiotic treatment, mechanical ventilation period, age, ARDS, parenteral nutrition, and gender did not show any independent risk factor for developing positive pulmonary samples for Candida species. The incidence of Candida isolation from pulmonary biopsies in critically ill mechanically ventilated, non-neutropenic patients who die is high (40%). However, the incidence of definite Candida pneumonia was 8%. We also found that Candida colonization is uniform throughout the different lung regions, and that the presence of Candida in respiratory samples, independently of quantitative cultures, is not a good marker of Candida pneumonia in critically ill, non-neutropenic, non-AIDS patients.