Paediatric Behçet's disease presenting with recurrent papillitis and episcleritis: a case report

<p>Abstract</p> <p>Introduction</p> <p>Behçet's disease is a chronic multisystem vasculitis characterized by mucocutaneous, articular, neurological, gastrointestinal and ophthalmological lesions. Ocular involvement is mainly represented by recurrent uveitis, especially posterior uveitis; however, iridocyclitis, retinal and choroidal vasculitis, optic neuritis and retinal vascular occlusion can also occur.</p> <p>Case presentation</p> <p>A 12-year-old Caucasian boy with a history of recurrent buccal aphthosis and nonspecific gastrointestinal symptoms was admitted to our hospital with blurred vision associated with acute episcleritis and papillitis. The patient's pathergy test was positive, suggesting a diagnosis of Behçet's disease. Corticosteroid and cyclosporine therapy was started, but further episodes were noted in both eyes. The patient was then switched to intravenous infliximab, with complete resolution of the inflammation after the second infusion.</p> <p>Conclusion</p> <p>Episcleritis and papillitis should be added to the list of uncommon manifestations of pediatric Behçet's disease. Infliximab is an effective, new therapeutic approach for Behçet's disease that is refractory to the conventional corticosteroid and immunosuppressive therapy.</p

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

Expression/Activation of PAR-1 in Airway Epithelial Cells of COPD Patients: Ex Vivo/In Vitro Study

The role of PAR-1 expression and activation was described in epithelial cells from the central and distal airways of COPD patients using an ex vivo/in vitro model. PAR-1 immunoreactivity was studied in epithelial cells from surgical specimens of the central and distal airways of COPD patients and healthy control (HC). Furthermore, PAR-1 expression and activation were measured in both the human bronchial epithelial cell line (16HBE) and normal human bronchial epithelial cells (NHBEs) exposed to cigarette smoke extract (CSE) (10%) or thrombin. Finally, cell proliferation, apoptosis, and IL-8 release were detected in stimulated NHBEs. We identified higher levels of PAR-1 expression/activation in epithelial cells from the central airways of COPD patients than in HC. Active PAR-1 increased in epithelial cells from central and distal airways of COPD, with higher levels in COPD smokers (correlated with pack-years) than in COPD ex-smokers. 16HBE and NHBEs exposed to CSE or thrombin showed increased levels of active PAR-1 (localized in the cytoplasm) than baseline conditions, while NHBEs treated with thrombin or CSE showed increased levels of IL-8 proteins, with an additional effect when used in combination. Smoking habits generate the upregulation of PAR-1 expression/activation in airway epithelial cells, and promoting IL-8 release might affect the recruitment of infiltrating cells in the airways of COPD patients

Functional Inactivation of pRB Results in Aneuploid Mammalian Cells After Release From a Mitotic Block

The widespread chromosome instability observed in tumors and in early stage carcinomas suggests that aneuploidy could be a prerequisite for cellular transformation and tumor initiation. Defects in tumor suppressors and genes that are part of mitotic checkpoints are likely candidates for the aneuploid phenotype. By using flow cytometric, cytogenetic, and immunocytochemistry techniques we investigated whether pRB deficiency could drive perpetual aneuploidy in normal human and mouse fibroblasts after mitotic checkpoint challenge by microtubule-destabilizing drugs. Both mouse and human pRB-deficient primary fibroblasts resulted, upon release from a mitotic block, in proliferating aneuploid cells possessing supernumerary centrosomes. Aneuploid pRB-deficient cells show an elevated variation in chromosome numbers among cells of the same clone. In addition, these cells acquired the capability to grow in an anchorage-independent way at the same extent as tumor cells did suggesting aneuploidy as an initial mutational step in cell transformation. Normal Mouse Embryonic Fibroblasts (MEFs) harboring LoxP sites flanking exon 19 of the Rb gene arrested in G2/M with duplicated centrosomes after colcemid treatment. However, these cells escaped the arrest and became aneuploid upon pRB ablation by CRE recombinase, suggesting pRB as a major component of a checkpoint that controls cellular ploidy

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

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

Demographic characteristics of the subjects.

*<p>p<0.01 vs ex-COPD.</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

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