Guanylyl cyclase activation reverses resistive breathing–induced lung injury and inflammation

Inspiratory resistive breathing (RB), encountered in obstructive lung diseases, induces lung injury. The soluble guanylyl cyclase (sGC)/cyclic guanosine monophosphate (cGMP) pathway is down-regulated in chronic and acute animal models of RB, such as asthma, chronic obstructive pulmonary disease, and in endotoxin-induced acute lung injury. Our objectives were to: (1) characterize the effects of increased concurrent inspiratory and expiratory resistance in mice via tracheal banding; and (2) investigate the contribution of the sGC/cGMP pathway in RB-induced lung injury. Anesthetized C57BL/6 mice underwent RB achieved by restricting tracheal surface area to 50% (tracheal banding). RB for 24 hours resulted in increased bronchoalveolar lavage fluid cellularity and protein content, marked leukocyte infiltration in the lungs, and perturbed respiratory mechanics (increased tissue resistance and elasticity, shifted static pressure–volume curve right and downwards, decreased static compliance), consistent with the presence of acute lung injury. RB down-regulated sGC expression in the lung. All manifestations of lung injury caused by RB were exacerbated by the administration of the sGC inhibitor, 1H-[1,2,4]oxodiazolo[4,3-]quinoxalin-l-one, or when RB was performed using sGCα1 knockout mice. Conversely, restoration of sGC signaling by prior administration of the sGC activator BAY 58-2667 (Bayer, Leverkusen, Germany) prevented RB-induced lung injury. Strikingly, direct pharmacological activation of sGC with BAY 58-2667 24 hours after RB reversed, within 6 hours, the established lung injury. These findings raise the possibility that pharmacological targeting of the sGC–cGMP axis could be used to ameliorate lung dysfunction in obstructive lung diseases

Possible role of glycosaminoglycans in the differential diagnostic approach of prostate adenocarcinoma

It is already known that chondroitin sulphate (CS) is accumulated around acini in benign prostatic hyperplasia (BPH) and in prostatic adenocarcinoma (PCa). In addition CS is a marker of poor prognosis in PCa. MCs are responsible directly or indirectly -by interacting with fibroblasts- for CS production, whilst CS influences MCs differentiation and degranulation. The aim of this study was to investigate CS and MCs’ alterations in BPH and PCa.The study comprised 67 prostate specimens (16,4% fine needle biopsies, 31,3% transurethral resections, 22,4% suprapubic prostatectomies and 29,9% radical prostatectomies) from patients aged 50-87 years old with benign prostatic hyperplasia (BPH)(46,3%) or prostatic adenocarcinoma (PCa)(53,7%). Multiple serial paraffin sections were stained for Alcian Blue pH 2.5, May Grünwald Giemsa and immunostained for the monoclonal mouse anti-chondroitin sulphate (anti-CS), clone CS-56 (Sigma-Aldrich, Inc.) and the monoclonal mouse anti-mast cell tryptase (anti- tryptase), clone AA1 (Neomarkers, Fremont, USA). By using a computer image analysis system (Image-Pro®Plus for Windows, Media Cybernetics, L.P.), we calculated the area of CS deposition (CS-MA/field, CS-MA%), the absorbance of CS deposition (CS-MOD), MCs’ mean number (MC-MN/field, MC-MN/Sd area), the mean diameter (MC-MD) and the mean area (MC-MA) of MCs’ cytoplasm. The results (mean ± SE) were analyzed by using non-parametric methods (SPSS 10.0 for Windows). All p values <0.05 were statistically significant.MCs’ mean number between acini was lower in BPH than in PCa (p<0,05). MCs’ cytoplasm size was smaller in BPH than in PCa, between acini (p<0,05) and around acini (p<0,05). In all cases, MCs’ mean number was higher around acini than between acini (p<0,001). In PCa group, MCs’ cytoplasm size around neoplastic acini was bigger than between neoplastic acini (p<0,05). When pT2-staged PCa cases were compared to the pT3-staged cases, the former had lower CS-MOD (p<0,05) and more MCs between acini (p<0,05) as well as around acini (p<0,05). In BPH group, CS- MOD showed weak negative relationship to MCs’ mean number around acini. In PCa group, the area of CS deposition showed weak positive relationship to the MCs’ cytoplasm size between acini, and CS-MOD showed weak negative relationship to the MCs’ cytoplasm size around acini. In neoplastic areas, MCs with anti-CS stained granules were found.CS and MCs show parallel alterations. MCs around acini are more responsible -than the MCs between acini- for CS production in prostatic diseases, and also MCs’ cytoplasm size seems to be a good marker of their activation.Είναι γνωστή η περιαδενική συσσώρευση της θειϊκής χονδροϊτίνης (CS) στην . καλοήθη υπερπλασία (ΒΡΗ) και στο αδενοκαρκίνωμα του προστάτη (PCa) καθώς και ο ρόλος της ως κακό προγνωστικό δείκτη στο PCa. Τα μαστοκύτταρα (MC) είναι υπεύθυνα άμεσα ή έμμεσα -μέσω των ινοβλαστών- για την παραγωγή της CS, ενώ η CS επηρεάζει τη διαφοροποίηση και την αποκοκκίωση των MC. Σκοπός της μελέτης αυτής είναι η διερεύνηση των μεταβολών της CS και των MC στην ΒΡΗ και στο PCa.Η μελέτη περιλαμβάνει 67 προστατικά παρασκευάσματα (16,4% διορθικές βιοψίες, 31,3% διουρηθρικές προστατεκτομές, 22,4% διακυστικές προστατεκτομές, 29,9% ριζικές προστατεκτομές) από ασθενείς ηλικίας 50-87 ετών με ΒΡΗ (46,3%) και PCa (53,7%). Πολλαπλές συνεχόμενες τομές παραφίνης βάφτηκαν με τις χρώσεις Alcian Blue pH 2.5 και May Grünwald Giemsa, και έγινε ανοσοϊστοχημεία για τα μονοκλωνικά αντισώματα mouse anti-chondroitin sulphate (anti-CS), clone CS-56 (Sigma-Aldrich, Inc.) και mouse anti-mast cell tryptase (anti-tryptase), clone AA1 (Neomarkers, Fremont, USA). Με ψηφιακή ανάλυση εικόνας έγινε προσδιορισμός της μέσης έκτασης του στρώματος που καταλαμβάνει η CS (CS-MA/field, CS- ΜΑ%), της μέσης οπτικής πυκνότητας της CS (CS-MOD), του μέσου αριθμού των MC (MC-MN/field, MC-MN/Sd area), της μέσης διαμέτρου και του μέσου εμβαδόν επιφάνειας (MC-MA) του κυτταροπλάσματος των MC. Τα αποτελέσματα (meaniSE) επεξεργάστηκαν στατιστικά με μη παραμετρικές μεθόδους (SPSS 10.0 for Windows). Όλες οι τιμές ρ<0,05 θεωρήθηκαν στατιστικά σημαντικές.Ο μέσος αριθμός των MC στο στρώμα μεταξύ των αδενίων ήταν μικρότερος στην ΒΡΗ απ’ό,τι στο PCa (ρ<0,05). Το μέγεθος των MCs ήταν μικρότερο στην ΒΡΗ απ’ό,τι στο PCa, στην περιοχή μεταξύ των αδενίων (ρ<0,05) αλλά και γύρω από τα αδένια (ρ<0,05). Σε όλα τα περιστατικά, ο μέσος αριθμός των MCs ήταν μεγαλύτερος γύρω από τα αδένια απ’ό,τι μεταξύ των αδενίων (ρ<0,001). Στην ομάδα PCa, το μέγεθος των MC γύρω από τα νεοπλασματικά αδένια ήταν μεγαλύτερο απ’ό,τι στην μεταξύ των νεοπλασματικών αδενίων περιοχή (ρ<0,05). Τα PCa σταδίου ρΤ2 σε σχέση με τα ρΤ3, είχαν μικρότερη CS-MOD (ρ<0,05) και περισσότερα MC τόσο γύρω από τα αδένια (ρ<0,05) όσο και μεταξύ των αδενίων (ρ<0,05). Στην ομάδα ΒΡΗ, η CS-MOD εμφανίζει ασθενή αρνητική συσχέτιση με τον αριθμό των MC γύρω από τα αδένια. Στην ομάδα PCa, η έκταση του στρώματος που καταλαμβάνει η CS εμφανίζει ασθενή θετική συσχέτιση με το μέγεθος των MC γύρω από τα αδένια, ενώ η CS-MOD εμφανίζει ασθενή αρνητική συσχέτιση με το μέγεθος των MC μεταξύ των αδενίων. Στις νεοπλασματικές περιοχές, εντοπίστηκαν MC με κοκκία σημασμένα με anti-CS.CS και MC εμφανίζουν παράλληλες μεταβολές. Τα MC του προστατικού στρώματος γύρω από τα αδένια φαίνεται να ευθύνονται περισσότερο -σε σχέση με τα MC του στρώματος μεταξύ των αδενίων- για την παραγωγή CS στις παθήσεις του προστάτη, και επιπλέον το μέγεθος τους φαίνεται να αντιπροσωπεύει το βαθμό ενεργοποίησής τους

Immunophenotype Heterogeneity in Nasal Glomangiopericytoma

Nasal glomangiopericytoma is rare. The immunophenotype is heterogeneous, more frequently smooth-muscle-actin and CD34-positive. We report expression patterns for several vascular-related proteins such as CD99, CD146, Bcl2, and WT1 as well as for treatment-related proteins such as mTOR and EGFR in a nasal glomangiopericytoma. The patient (woman, 86 years) presented with a left nasal tumefaction. The resected specimen (1.5-cm) showed a glomangiopericytoma. Tumor cells expressed smooth-muscle-actin, CD31, CD34, and progesterone receptor. They also expressed the vascular-cell-related proteins Bcl2, CD99, CD146, and WT1, as well as mTOR and EGFR. Nasal glomangiopericytomas show immunohistochemical heterogeneity for vascular-related markers, suggesting a possible extensive pericytic differentiation. The expression of potential targets for drug treatments such as mTOR and EGFR may impact on the clinical follow-up of these tumors occurring at advanced ages, which may require complex surgery

Highly Selective Endothelin-1 Receptor A Inhibition Prevents Bleomycin-Induced Pulmonary Inflammation and Fibrosis in Mice

Background: Pulmonary fibrosis is a chronic disease, which progressively leads to respiratory failure and ultimately death. Endothelin-1 (ET-1), a vasoconstrictor secreted by endothelial cells, promotes vasoconstriction by activation of its receptors A and B. Objectives: We addressed the role of highly selective ET-1 receptor A (ETA) inhibition in the pathogenesis of experimental pulmonary fibrosis by bleomycin (BLM). Methods: BLM sulfate (2 U/mL) or saline was intratracheally administered to C57/Bl6 mice (4 groups; n = 5-11/group). Pretreatment with the highly selective ETA receptor inhibitor sitaxentan (15 mg/kg/day) was started 1 day prior to BLM injection and continued for the duration of the experiment. Lung mechanics were assessed prior to sacrifice at days 7, 14, and 21 after BLM, followed by procurement of bronchoalveolar lavage fluid (BALF), blood, and lung tissue samples. Results: Time-dependent effects of BLM exposure included decreased static compliance and increased lung elastance, airspace inflammation and microvascular permeability, histological acute lung injury and fibrosis, and lung collagen deposition. Pretreatment with highly selective ETA receptor inhibitor had no adverse effect on control mice but improved lung mechanics and lung injury score in addition to decreasing BALF pleocytosis, protein content, and collagen deposition in BLM-treated mice. Mortality from BLM reached 40% and occurred primarily during the inflammatory stage of the model but was abrogated by sitaxentan pretreatment. Conclusions: We conclude that in our BLM-induced pulmonary fibrosis model, prophylactic highly selective ETA inhibition improves survival, preserves lung function, attenuates lung injury, and reduces collagen deposition. (c) 2017 S. Karger AG, Base