Beta defensin-2 is reduced in central but not in distal airways of smoker COPD patients

Background: Altered pulmonary defenses in chronic obstructive pulmonary disease (COPD) may promote distal airways bacterial colonization. The expression/activation of Toll Like receptors (TLR) and beta 2 defensin (HBD2) release by epithelial cells crucially affect pulmonary defence mechanisms. Methods: The epithelial expression of TLR4 and of HBD2 was assessed in surgical specimens from current smokers COPD (s-COPD; n = 17), ex-smokers COPD (ex-s-COPD; n = 8), smokers without COPD (S; n = 12), and from non-smoker non-COPD subjects (C; n = 13). Results: In distal airways, s-COPD highly expressed TLR4 and HBD2. In central airways, S and s-COPD showed increased TLR4 expression. Lower HBD2 expression was observed in central airways of s-COPD when compared to S and to ex-s-COPD. s-COPD had a reduced HBD2 gene expression as demonstrated by real-time PCR on micro-dissected bronchial epithelial cells. Furthermore, HBD2 expression positively correlated with FEV1/FVC ratio and inversely correlated with the cigarette smoke exposure. In a bronchial epithelial cell line (16 HBE) IL-1β significantly induced the HBD2 mRNA expression and cigarette smoke extracts significantly counteracted this IL-1 mediated effect reducing both the activation of NFkB pathway and the interaction between NFkB and HBD2 promoter. Conclusions: This study provides new insights on the possible mechanisms involved in the alteration of innate immunity mechanisms in COPD. © 2012 Pace et al

Significance of the effective remnant liver volume in major hepatectomies

The aim of this study is to identify the minimum safe amount of effective remnant liver volume (ERLV) in patients undergoing a major hepatectomy. Thirty-eight consecutive major hepatectomies (resection of > or = 3 Couinaud segments) performed between July 1999 and March 2004 in which a frozen section liver biopsy was obtained were included. No patient had chronic viral hepatitis, cirrhosis, or cholestasis. The total liver volume (TLV) was calculated using the Vauthey formula, and the postsurgical liver volume (PSLV) was derived by subtracting the estimated volume of liver resected from the TLV. The PSLV minus the percentage of macrovesicular steatosis as nonfunctional liver was defined as the effective remnant liver volume (ERLV). Three groups of ERLV/TLV ratios (60%) were correlated with liver resection type, mortality, complications, intraoperative blood transfusions, operative time, length of hospitalization, and mean value of liver function tests in the first 5 postoperative days. Comparisons between clinical parameters were performed by Pearson chi2 test. There was significant correlation between ERLV/TLV ratios and surgical resection type (P < 0.001), early postoperative mortality (P < 0.01), and complications (P < 0.003). The ERLV/TLV ratio may be a useful predictor of surgical outcome after major hepatectomy

Significance of the effective remnant liver volume in major hepatectomies

The aim of this study is to identify the minimum safe amount of effective remnant liver volume (ERLV) in patients undergoing a major hepatectomy. Thirty-eight consecutive major hepatectomies (resection of > or = 3 Couinaud segments) performed between July 1999 and March 2004 in which a frozen section liver biopsy was obtained were included. No patient had chronic viral hepatitis, cirrhosis, or cholestasis. The total liver volume (TLV) was calculated using the Vauthey formula, and the postsurgical liver volume (PSLV) was derived by subtracting the estimated volume of liver resected from the TLV. The PSLV minus the percentage of macrovesicular steatosis as nonfunctional liver was defined as the effective remnant liver volume (ERLV). Three groups of ERLV/TLV ratios (60%) were correlated with liver resection type, mortality, complications, intraoperative blood transfusions, operative time, length of hospitalization, and mean value of liver function tests in the first 5 postoperative days. Comparisons between clinical parameters were performed by Pearson chi2 test. There was significant correlation between ERLV/TLV ratios and surgical resection type (P < 0.001), early postoperative mortality (P < 0.01), and complications (P < 0.003). The ERLV/TLV ratio may be a useful predictor of surgical outcome after major hepatectomy

Cystic echinococcosis of the liver and lung treated by radiofrequency thermal ablation: An ex-vivo pilot experimental study in animal models

AIM: To evaluate radiofrequency thermal ablation (RTA) for treatment of cystic echinococcosis in animal models (explanted organs)

Expression of HBD2 in distal and in central airways.

<p>Immunohistochemistry for HBD2 in distal (A) and in central airways (B) from surgical samples of Controls (n = 13), S (n = 12), s-COPD (n = 17) and ex-s-COPD (n = 8) subjects. Cells were stained with an anti-HBD2 antibody. Negative control were performed using rabbit immunoglobulins negative control (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033601#s2" target="_blank">materials and methods</a> for details). <b>A</b>) Individual counts for the number of positive epithelial cells/mm basement membrane in distal airways. Horizontal bars represent median values. * p<0.05 values in figure represent Mann-Whitney U test analyses. <b>B</b>) Individual counts for the number of positive epithelial cells/mm basement membrane in central airways. Horizontal bars represent median values. * p<0.05 values in figure represent Mann-Whitney U test analyses.</p

Expression of HBD2 m-RNA in central airways.

<p>HBD2 m-RNA expression was assessed by Real time PCR in microdissected bronchial epithelium from s-COPD (n = 3) and from ex-s-COPD (n = 3) (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033601#s2" target="_blank">materials and methods</a> for details). <b>A</b>) Representative images showing the bronchial epithelium before (on the left) and after (on the right) laser microdissection (LMD). <b>B</b>) Expression of HBD2 m-RNA in microdissected bronchial epithelium. GAPDH gene expression was used as endogenous control for normalization. Relative quantitation of mRNA was carried out with comparative CT method. (mean±SD). * p<0.05.</p

Effects of CSE in bronchial epithelial cells (16-HBE).

<p>16-HBE cells were cultured in the presence and in the absence of IL-1 β and of CSE (10%) (n = 3) (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033601#s2" target="_blank">materials and methods</a> for details). <b>A</b>) Expression of HBD2 m-RNA in 16-HBE by real time PCR. GAPDH gene expression was used as endogenous control for normalization. Relative quantitation of mRNA was carried out with comparative CT method. (mean±SD). * p<0.05 versus baseline. ** p<0.05 versus IL-1 beta. <b>B</b>) Representative experiment (one out of three experiments) showing the expression of HBD2 protein in 16-HBE by flow cytometry. The expression of HBD2 is expressed as percentage of HBD2 positive cells. <b>C</b>) Evaluation of p-IkBa or t-IkBa by western blot analysis. Membranes were then stripped and incubated with goat polyclonal anti–ß-actin. Representative western blot analysis (one out of three experiments). Lane1 = baseline; lane 2 = CSE 10%;lane 3 = IL1 beta; lane 4 = CSE+IL1 beta. <b>D</b>) ChiP assay using anti-NFkB antibody and PCR using primers (forward 5′-GGTGTGAATGGAAGGAACTCA-3′ reverse 5′-TTCAGCTCCTGGGGATGATAC-3′) spanning the promoter region of HBD2 gene were performed (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033601#s2" target="_blank">Materials and Methods</a> for details) One out of two experiments is shown. Lane 1 = DNA marker; Lane 2 = baseline; lane 3 = CSE 10%; lane 4 = IL1 β; lane 5 = CSE+IL1 β.</p

TLR4 immunostaining in distal and in central airways.

<p><b>A</b>) Representative negative control and representative TLR4 immunostaining (red stain) in distal airways of a Control, of a Smoker, of a s-COPD and of an ex-s-COPD. <b>B</b>) Representative negative control and representative TLR4 immunostaining (red stain) in central airways of a Control, of a Smoker, of a s-COPD and of an ex-s-COPD. For central airways a particular from a 400× magnification was selected and showed.</p

HBD2 immunostaining in distal and in central airways.

<p><b>A</b>) Representative negative control and representative HBD2 immunostaining (red stain) in distal airways of a Control, a Smoker, a s-COPD and an ex-s-COPD. <b>B</b>) Representative negative control and representative HBD2 immunostaining (red stain) in central airways of a Control, of a Smoker, of a s-COPD and of ex-s-COPD. For central airways a particular from a 400× magnification was selected and showed.</p

Correlations between the expression of HBD2 in central airways and functional parameters.

<p>The expression of HBD2 in central airways of Controls (n = 13), S (n = 12), s-COPD (n = 17) and ex-s-COPD (n = 8) was correlated with FEV1/FVC ratio (<b>A</b>) and packs/year (<b>B</b>) by Spearman Correlation test.</p