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Tissue inhibitor of metalloproteinase-1 (TIMP-1) is an important regulator of extracellular matrix turnover that has been traditionally regarded as a potential tumor suppressor owing to its inhibitory effects of matrix metalloproteinases. Intriguingly, this interpretation has been challenged by the consistent observation that increased expression of TIMP-1 is associated with poor prognosis in virtually all cancer types including lung cancer, supporting a tumor-promoting function. However, how TIMP-1 is dysregulated within the tumor microenvironment and how it drives tumor progression in lung cancer is poorly understood. We analyzed the expression of TIMP-1 and its cell surface receptor CD63 in two major lung cancer subtypes: lung adenocarcinoma (ADC) and squamous cell carcinoma (SCC), and defined the tumor-promoting effects of their interaction. We found that TIMP-1 is aberrantly overexpressed in tumor-associated fibroblasts (TAFs) in ADC compared to SCC. Mechanistically, TIMP-1 overexpression was mediated by the selective hyperactivity of the pro-fibrotic TGF-β1/SMAD3 pathway in ADC-TAFs. Likewise, CD63 was upregulated in ADC compared to SCC cells. Genetic analyses revealed that TIMP-1 secreted by TGF-β1-activated ADC-TAFs is both necessary and sufficient to enhance growth and invasion of ADC cancer cells in culture, and that tumor cell expression of CD63 was required for these effects. Consistently, in vivo analyses revealed that ADC cells co-injected with fibroblasts with reduced SMAD3 or TIMP-1 expression into immunocompromised mice attenuated tumor aggressiveness compared to tumors bearing parental fibroblasts. We also found that high TIMP1 and CD63 mRNA levels combined define a stronger prognostic biomarker than TIMP1 alone. Our results identify an excessive stromal TIMP-1 within the tumor microenvironment selectively in lung ADC, and implicate it in a novel tumor-promoting TAF-carcinoma crosstalk, thereby pointing to TIMP-1/CD63 interaction as a novel therapeutic target in lung cancer.

Fibrillar collagens are the most abundant extracellular matrix components in nonesmall cell lung cancer (NSCLC). However, the potential of collagen fiber descriptors as a source of clinically relevant biomarkers in NSCLC is largely unknown. Similarly, our understanding of the aberrant collagen organization and associated tumor-promoting effects is very scarce. To address these limitations, we identified a digital pathology approach that can be easily implemented in pathology units based on CT-FIRE software (version 2; https://loci.wisc.edu/software/ctfire) analysis of Picrosirius red (PSR) stains of fibrillar collagens imaged with polarized light (PL). CT-FIRE settings were pre-optimized to assess a panel of collagen fiber descriptors in PSR-PL images of tissue microarrays from surgical NSCLC patients (106 adenocarcinomas [ADC] and 89 squamous cell carcinomas [SCC]). Using this approach, we identified straightness as the single high-accuracy diagnostic collagen fiber descriptor (average area under the curve ¼ 0.92) and fiber density as the single descriptor consistently associated with a poor prognosis in both ADC and SCC independently of the gold standard based on the TNM staging (hazard ratio, 2.69; 95% CI, 1.55-4.66; P

Background The TGF-β1 transcription factor SMAD3 is epigenetically repressed in tumour-associated fibroblasts (TAFs) from lung squamous cell carcinoma (SCC) but not adenocarcinoma (ADC) patients, which elicits a compensatory increase in SMAD2 that renders SCC-TAFs less fibrotic. Here we examined the effects of altered SMAD2/3 in fibroblast migration and its impact on the desmoplastic stroma formation in lung cancer. Methods We used a microfluidic device to examine descriptors of early protrusions and subsequent migration in 3D collagen gels upon knocking down SMAD2 or SMAD3 by shRNA in control fibroblasts and TAFs. Results High SMAD3 conditions as in shSMAD2 fibroblasts and ADC-TAFs exhibited a migratory advantage in terms of protrusions (fewer and longer) and migration (faster and more directional) selectively without TGF-β1 along with Erk1/2 hyperactivation. This enhanced migration was abrogated by TGF-β1 as well as low glucose medium and the MEK inhibitor Trametinib. In contrast, high SMAD2 fibroblasts were poorly responsive to TGF-β1, high glucose and Trametinib, exhibiting impaired migration in all conditions. Conclusions The basal migration advantage of high SMAD3 fibroblasts provides a straightforward mechanism underlying the larger accumulation of TAFs previously reported in ADC compared to SCC. Moreover, our results encourage using MEK inhibitors in ADC-TAFs but not SCC-TAFs.

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.

7 páginas, 4 tablas., Finding alternatives to deal with the lack of feed during the dry period is a challenge for animal research in tropical areas. The effect of age of regrowth (60, 120 or 180 days), chemical composition and secondary metabolites on Leucaena leucocephala digestibility was evaluated. A randomized block design with six replicates was used. The statistical procedures included analysis of variance and correlation and regression analysis. Multiple linear equations between in vitro and in situ digestibility, as dependent variables, and the age of regrowth, NDF, ADF, lignin, Ca, Si, NDF/N, ADF/N, tannins, condensed tannins and saponins, as explanatory variables, were established. An increase in cell wall content and its components with age of regrowth, and a variable response in mineral concentrations were observed. The linear equations relating digestibility with the age of regrowth, NDF, ADF and Si (R2[0.78) or with the secondary metabolites tannins, condensed tannins and saponins (R2[ 0.79) were statistically significant (p\0.05) explaining a high proportion of variance. The regrowth age, cell wall components, polyphenol compounds, saponins contents determined the forage digestibility and these aspects must be taken into consideration to estimate the nutritional value of Leucaena roughage to be used as feedstuff for animals.

Large cell carcinoma (LCC) is a rare and aggressive lung cancer subtype with poor prognosis and no targeted therapies. Tumor-associated fibroblasts (TAFs) derived from LCC tumors exhibit premature senescence, and coculture of pulmonary fibroblasts with LCC cell lines selectively induces fibroblast senescence, which in turn drives LCC cell growth and invasion. Here we identify MMP1 as overexpressed specifically in LCC cell lines, and we show that expression of MMP1 by LCC cells is necessary for induction of fibroblast senescence and consequent tumor promotion in both cell culture and mouse models. We also show that MMP1, in combination with TGF-β1, is sufficient to induce fibroblast senescence and consequent LCC promotion. Furthermore, we implicate PAR-1 and oxidative stress in MMP1/TGF-β1-induced TAF senescence. Our results establish an entirely new role for MMP1 in cancer, and support a novel therapeutic strategy in LCC based on targeting senescent TAFs., This work was further supported by grants from the Agencia Estatal de Investigaci´on (AEI/FEDER) (PI13/02368, SAF2016-79527-R and PID2019-110944RB-I00 (AEI/ 10.13039/501100011033) to JA, PI16/ 00890 to NR), Fundaci´o Privada Cellex (to JA), National Institutes of Health (grant R01 GM132100 to ER), Generalitat de Catalunya (AGAUR SGR661 and CERCA Programme to JA), Junta Provincial de Barcelona de l’Associaci´o Espanyola Contra el C`ancer (AECC-B16-917 to JA), Italian Ministry of Health (RF-2016-02362946 to LR), Italian Association for Cancer Research (AIRC-IG21431 to LR), Sociedad Espa˜nola de Neumología y Cirugía Tor´acica – SEPAR (SEPAR 437 to NR), COST Action (PROTEOSTASIS BM1307), and by fellowships from Ciˆencia sem Fronteiras CNPq (to RI), and Universitat de Barcelona/beca APIF (to PD).

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).

10 páginas, 4 tablas., This experiment has examined the effects of age of regrowth (60, 120 or 180 days) and season (dry or rainy period) on the nutritional quality of roughage from Erythrina variegata harvested in the Cauto Valley, Cuba. A randomized block design with three replicates was used. Proximate and mineral composition, in vitro and in situ dry matter digestibility were evaluated, in addition to the secondary metabolites (tannins, phenols, verbascose, stachyose, raffinose, flavonoids, triterpenes and steroids) at different ages of regrowth, in both seasons of the year. There was a significant interaction between treatments for most of the nutritional components or digestibility coefficients analyzed, although the pattern followed with age of regrowth was similar in both seasons. The gas production increased with the age of regrowth and the higher value was obtained during the dry season, while the lag phase was longer in the rainy season. The results of this study show the marked effect of climatic factors on the nutritive quality of E. variegata. In general, as maturity was increased the concentrations of secondary metabolites also increased, but only condensed tannins were close to the limits for which ruminal fermentations can be affected. All these aspects should be considered in this species when it is used in animal feeding.

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.)

The present work was developed with the objective of evaluating the effect of the regrowth age on the chemical composition, digestibility and polyphenol content of the Jatropha curcas in an area of Cauto Valley. Was used piece of ground of Jatropha curcaswith two years of establishment. A randomized block design with six replicas was used. The treatments were regrowth ages of60, 120 and 180 days. At the beginning of each seasonal period a homogeneity cut was made at 1 m above the ground level. Fromthere the sampling was carried out in 10 random plants eliminating the edge effect in an area of 0.5 ha, according to the treatments.The chemical composition of Jatropha curcas the dry matter content, DNF, ADF, ADL, increase with age with its best results at 180 days with 28.82 ; 51.53 ; 33.44 and 20.22 % ; while the CP and cellular content decrease with its highest results at 60 days 27.78 and58.79 %. The minerals showed a very variable behavior. The highest energy contribution and digestibility percentage at 60 days of age with a decrease of 11.04 ; 7.40 ; 8.90 % ; 1.58 and 1.11 MJ / kg. Polyphenolic compounds increase with age with the best results at180 days of age with 11.26, 21.10, 113.93, 111.49 and 2.46 g / Kg, respectively ; for total tannins, total phenols, TCT, TBCT and FTC. The results of the present investigation evidenced the marked effect of climatic factors and age on the quality of the Jatropha curcas, as the maturity of the forage progressed, its nutritional contribution decreased with a decrease in the CP, the nutrient content, digestibility and energy, as well as an increase in the fibrous fraction and the content of polyphenolic compounds.

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.

Background In the context of complex diseases like psoriatic arthritis (PsA), in which patients are often followed by different professionals, it is important for health professionals, providers and patients to have tools for delivering and demanding optimal care. One way to organise and evaluate health care quality is by the use of validated standards of care and quality indicators. Objectives To develop nationally accepted standards of care and quality indicators for care in PsA. Methods Qualitative methodology was followed that included: 1)Two focus groups (one with patients with PsA and another with non-rheumatologists specialists involved in the care of patients with PsA; 2) A narrative literature review of published documents related to the quality of care in PsA (including the QUANTUM Report1); 3) A nominal group meeting in which 15 expert rheumatologists generated and consensuated, a series of quality criteria as well as formulas or quantifiable objective measures to evaluate them; 4) A Delphi to establish the feasibility, priority and agreement with the quality criteria; 5) A final generation of standards of care and their attributes (including quality indicators). A descriptive analysis of the results was carried out. Results A total of 59 standards of care were generated, 19 of mandatory compliance, grouped into 4 blocks according to specific objectives: 1) Early diagnosis (n=6); 2) optimising the management of the disease (n=26), 3) multidisciplinary collaboration (n=9); 4) monitoring improvement (n=18). To assess the compliance of these standards of care, in many cases the medical records will be reviewed. Other sources will be the memory of the service and hospital and bibliographic databases. Regarding to the level compliance, for some of the standards of care this is yes/no, for others the compliance will range are from 50% to 100%, and in this range for many this will be by 80%. Conclusions This set of standards of care should help improve quality of care in APs patients. Reference [1] Favier G, Gladman DD, Merola JF, Armstrong AW, Boehncke WH, Helliwell PS. Benchmarking Care in Psoriatic Arthritis – The QUANTUM Report: A Report from the GRAPPA 2016 Annual Meeting. J Rheumatol. 2017;44(5):674–8. Disclosure of Interest None declared

©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).

Introduction: In pleural thickening, study ultrasound as well as serving as a guide, provides useful information to predict the nature of the same. We consider that it is possible to describe the set of echographic findings that define pleural thickening due to extrapleural fat (EPF). These findings, that we will call Extrapleural Fat Signs (EPFS), are: homogeneity, edges regularity, hypo or anecogenicity plus hyperechogenic lower border. The objective of the study is to determine whether such EPFS are indicative of EPF or not. Method: Since 2009, we performed at two third level hospitals, a transversal, prospective study, that analyzed thoracic ultrasounds with pleural thickening. We collected ultrasound characteristics of pleural thickening. All lesions were studied by CT and checked the etiology of no EPF thickening with biopsy. We considered as EPF any lesion with negative Hounsfield units in CT. Results: 66 patients were studied;obtaining the following results:27(41%)malignant lesions and 34(52%)benign lesions. In 5(7%)of the patients studied, diagnosis was not possible. The ultrasound diagnosed 25 lesions suggesting EPF, which were confirmed in all cases by CT, implying 100% Sensitivity. No EPF lesions were observed in CT that did not have a compatible ultrasound resulting in a negative predictive value of 100%.None of the lesions oriented as malignant or benign lesions different from EPF showed EPFS on ultrasound, implying 100% Specificity. Conclusions: 1.Detection using thoracic ultrasound of EPFS is a sensitive and specific method with a high NPV for the identification of pleural thickening compatible with EPF.2. The presence of EPFS would allow us to avoid performing invasive techniques such as pleural biopsy.

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.

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. To address this question we conducted a retrospective multicentric study of the prognostic value of the standard markers of activated fibroblasts. For this purpose, 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. 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. 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. Moreover it supports that antifibrotic drugs aiming to target tumor fibrosis may be an effective therapeutic approach to improve survival in NSCLC. Citation Format: Jordi Alcaraz, Josep-Lluís Carrasco, Laura Millares, Iuliana-Cristiana Benchea, Francisco-Javier Fernández, Anabel Martínez, María José Pajares, Julio Sánchez de Cos, Ramon Rami-Porta, Luis Seijo, Josep Ramírez, Noemí Reguart, Esther Barreiro, Eduard Monsó. Stromal markers of activated tumor associated fibroblasts predict poor survival and are associated with necrosis in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3160.

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.

Resumen Objetivo Analizar el significado de la presencia de linfocitos en las muestras de puncion transbronquial aspirativa (PTA) de adenopatias mediastinicas (AM) en pacientes con sospecha de cancer de pulmon (CP). Metodos Estudio observacional retrospectivo que evalua el valor predictivo negativo (VPN) de las muestras de PTA con evidencia de linfocitos pero sin celulas atipicas. Resultados Se realizaron 266 PTA a 252 pacientes con AM patologicas. En 115 PTA se evidencio la presencia de metastasis ganglionares (43%), y 94 (35%) fueron consideradas como no valorables (ausencia de material citologico evaluable o presencia exclusiva de celulas epiteliales bronquiales). De las 57 muestras de PTA restantes que contenian linfocitos sin atipias (21%), en 15 no se pudo confirmar el diagnostico; en 32 se confirmo mediante tecnicas diagnosticas alternativas y en 10 mediante seguimiento clinico-radiologico. El VPN de las 32 muestras confirmadas con tecnicas diagnosticas alternativas fue del 84% y descendio al 76% cuando se incluyeron las 10 PTA en las que se disponia de seguimiento clinico-radiologico. Conclusiones La presencia de linfocitos sin atipias en la muestra de PTA no excluye la invasion neoplasica del ganglio analizado.

7 páginas, 4 tablas., Ensiled king grass (Cenchrus purpureus (Schumach.) Morrone) is commonly used as feed in tropical ruminant production. However, ruminant performance can be limited by low nitrogen (N) content in tropical grass silage. A mixed feed of legume–king grass silage may be an option to improve ruminant production. We investigated the effects of feeding an ensiled mixture of king grass forage and foliage of a browse tree legume, either leucaena (Leucaena leucocephala (Lam.) de Wit) or gliricidia (Gliricidia sepium (Jacq.) Walp), on dry matter (DM) intake, DM digestibility, organic matter (OM) digestibility and N balance of Pelibuey sheep. Twelve male lambs 9–11 months old were blocked by initial bodyweight (33 ± 3 kg, mean ± s.d.) into three groups of four lambs and randomly assigned to one of three feed treatments: king grass silage (KS); mixed king grass and leucaena silage (KLS); and mixed king grass and gliricidia silage (KGS). Results showed that DM and OM were similar among silages. The N content was higher (P < 0.001) in KLS and KGS than in KS. DM intake and DM digestibility were higher (P < 0.001) in KLS and KGS than in KS. OM digestibility was higher (P < 0.001) in KLS than in KS. Nitrogen intake, faecal N, urine volume, urinary N, digested N, N digestibility, N retention and retained N : N intake ratio from KLS and KGS treatments were higher (P < 0.05) than from KS. A lower (P < 0.01) urinary N : N intake ratio was detected from KGS and KLS than from KS. This study demonstrated that, compared with silage made from king grass alone, legume–king grass silage mixtures can improve intake and nutrient digestibility in Pelibuey lambs. Further, N retention can be enhanced when legume–king grass silages are used as feed compared with king grass silage alone., The Startup Research Fund offered by Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, to Long Cheng is acknowledged to support this international collaboration. We thank Miss Jiawen Li (Chengdu University of Information Technology) for useful discussion and help with formatting the manuscript for submission.

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.

Background: Little evidence is available concerning the impact of virtual microscopy (VM) in the undergraduate teaching of pathology. We aimed: (1) to determine the impact in student scores when moving from conventional microscopy (CM) to VM; (2) to assess the students′ impressions and changes in study habits regarding the impact of this tool. Methods: We evaluated two groups taking the discipline of pathology in the same course, one using CM and the other VM. The same set of slides used in the CM classes was digitized in a VENTANA iScan HT (Roche Diagnostics, Sant Cugat, Spain) at ×20 and observed by the students using the Virtuoso viewer (Roche Diagnostics). We evaluated the skill level reached by the students with an online test. A voluntary survey was undertaken by the VM group to assess the students′ impressions regarding the resource. The day and time of any accession to the viewer were registered. Results: There were no differences between the two groups in their marks in the online test (mean marks for the CM and the VM groups: 9.87 ± 0.34 and 9.86 ± 0.53, respectively; P = 0.880). 86.6% of the students found the software friendly, easy-to-use and effective. 71.6% of the students considered navigation easier with VM than with CM. The most appreciated feature of VM was the possibility to access the images anywhere and at any time (93.3%). 57.5% of the accesses were made on holidays and 41.9% later than 6:00 pm. Conclusions: Virtual microscopy can effectively replace the traditional methods of learning pathology, providing mobility and convenience to medical students.

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 …

Cigarette smoking has been associated with alterations in the structure and endothelial function of pulmonary arteries. Nitric oxide (NO) and endothelin-1 are endothelium-derived mediators with opposite effects on vascular tone and cell growth. To investigate whether cigarette smoking could induce changes in the synthesis of these mediators in pulmonary arteries, we compared the expression of both endothelial NO synthase (eNOS) and endothelin-1 in the lungs of smokers with that in nonsmokers. Lung tissue samples of 23 smokers and nine nonsmokers were studied. Expression of eNOS and endothelin-1 in pulmonary artery endothelium was evaluated by immunohistochemistry. In protein extracts of lung tissue, the content of eNOS protein was assessed by Western blot analysis and that of endothelin-1 by radioimmunoassay. The immunohistochemical expression of eNOS in arterial endothelium and the eNOS protein content in lung tissue were lower in the smokers than in the nonsmokers. No differences were shown in cell expression and protein content of endothelin-1 between both groups. We conclude that cigarette smoking is associated with reduced expression of eNOS in pulmonary arteries. The diminished synthesis of nitric oxide may contribute to the alterations in the structure and endothelial function of pulmonary vessels in cigarette-smoke-induced respiratory disease.

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.