Use of an Amplatzer Device for Endoscopic Closure of a Large Bronchopleural Fistula following Lobectomy for a Stage I Squamous Cell Carcinoma

Bronchopleural fistulas can occur as a rare but severe complication after pulmonary resection. Established guidelines for the proper treatment of patients with bronchopleural fistulas do not exist. Apart from attempts to close the fistula, emphasis is placed on preventive measures, early treatment with antibiotics, drainage of the empyema and aggressive nutritional and rehabilitative support. For inoperable patients, endoscopic procedures are the only therapeutic option. Unfortunately, large (>8 mm) or central bronchopleural fistulas are usually not suitable for such endoscopic management. Recently, some groups have published a few case reports about a novel technique for the endobronchial closure of bronchopleural fistulas, using an Amplatzer device, originally designed for transcatheter closure of cardiac septal defects. We applied the same technique as a life-saving treatment in a ventilated patient who was considered inoperable due to a high oxygen need. The operation was successful. The patient could be weaned from ventilation and was eventually discharged from the hospital to a rehabilitation facility several weeks after the insertion of the device. Until now, endoscopic techniques have only been useful for the treatment of small, peripheral, bronchopleural fistulas and even then only as a bridge to surgery in high-risk surgical patients. In this case report, we demonstrate that the use of an Amplatzer device can expand the importance of endoscopic techniques in the treatment of bronchopleural fistulas. An Amplatzer device, for endobronchial closure, can indeed be administered for large and central bronchopleural fistulas. Moreover, it can be considered as a definite alternative to surgery in inoperable patients

Functional respiratory imaging to assess the interaction between systemic roflumilast and inhaled ICS/LABA/LAMA

Wim Vos,1 Bita Hajian,2 Jan De Backer,1 Cedric Van Holsbeke,1 Samir Vinchurkar,1 Rita Claes,2 Annemie Hufkens,2 Paul M Parizel,3 Lieven Bedert,4 Wilfried De Backer2 1FLUIDDA nv, Groeningenlei, Kontich, 2Department of Respiratory Medicine, 3Department of Radiology, University Hospital Antwerp, Wilrijkstraat, Edegem, 4Department of Respiratory Medicine, ZNA Middelheim Hospital, Antwerp, Belgium Background: Patients with COPD show a significant reduction of the lobar hyperinflation at the functional residual capacity level in the patients who improved >120 mL in forced expiratory volume in 1 second (FEV1) after 6 months of treatment with roflumilast in addition to inhaled corticosteroids (ICSs)/long-acting beta-2 agonists (LABAs)/long-acting muscarinic antagonists (LAMAs).Methods: Functional respiratory imaging was used to quantify lobar hyperinflation, blood vessel density, ventilation, aerosol deposition, and bronchodilation. To investigate the exact mode of action of roflumilast, correlations between lobar and global measures have been tested using a mixed-model approach with nested random factors and Pearson correlation, respectively.Results: The reduction in lobar hyperinflation appears to be associated with a larger blood vessel density in the respective lobes (t=−2.154, P=0.040); lobes with a higher percentage of blood vessels reduce more in hyperinflation in the responder group. Subsequently, it can be observed that lobes that reduce in hyperinflation after treatment are better ventilated (t=−5.368, P<0.001). Functional respiratory imaging (FRI)-based aerosol deposition showed that enhanced ventilation leads to more peripheral particle deposition of ICS/LABA/LAMA in the better-ventilated areas (t=2.407, P=0.024). Finally, the study showed that areas receiving more particles have increased FRI-based bronchodilation (t=2.564, P=0.017), leading to an increase in FEV1 (R=0.348, P=0.029).Conclusion: The study demonstrated that orally administered roflumilast supports the reduction of regional hyperinflation in areas previously undertreated by inhalation medication. The local reduction in hyperinflation induces a redistribution of ventilation and aerosol deposition, leading to enhanced efficacy of the concomitant ICS/LABA/LAMA therapy. FRI appears to be a sensitive tool to describe the mode of action of novel compounds in chronic obstructive pulmonary disease. Future studies need to confirm the enhanced sensitivity and the potential of FRI parameters to act as surrogates for clinically relevant, but more difficult to measure, end points such as exacerbations. Keywords: modeling, physiology, ventilation, aerosol distribution, FR

HCV upregulates ADRP expression in Huh-7 cells.

<p><b>(A)</b> Representative immunoblot of ADRP, core and β-actin in Huh-7 cells untransfected or transfected with Jc1 full length RNA (HCV). <b>(B)</b> Graph represents ADRP protein quantifications of at least three independent experiments. <b>(C)</b> ADRP mRNA levels were assessed by RT-qPCR.</p

Effect of ADRP overexpression on HCV particle production.

<p>GFP and ADRP transduced cells were infected by Jc1 viral particles. Intracellular proteins, RNA and particles were harvested 48 h post infection to assess the level of HCV core protein expression by immunoblot <b>(A,B)</b>, the relative number of intracellular HCV RNA copies by RT-qPCR <b>(C)</b> and the intracellular particle infectivity by infecting naive Huh-7.5 cells and calculating the TCID/50 <b>(D)</b>. In parallel, supernatants were collected to determine the relative number of extracellular HCV RNA copies by RT-qPCR <b>(E)</b> and the extracellular particle infectivity by infecting naive Huh-7.5 cells and calculating the TCID₅₀/ml <b>(F).</b></p

ADRP overexpression, but not ADRP silencing, modifies morphology and lipid content of lipid droplets.

<p><b>(A)</b> ORO staining of ADRP overexpressing (b) or silenced (d) cells Huh-7 cells compared to their respective controls (GFP, a and siCont, c). <b>(B,C)</b> Determination of morphometric features of LDs in control vs. ADRP overexpressing or silenced cells. LD number per cell <b>(B)</b> and total surface of LDs per cell <b>(C)</b> were quantified using the Metamorph software. Cellular TG <b>(D)</b> and CE and free cholesterol <b>(E)</b> contents normalized by the total protein amount are shown. <b>(F)</b> mRNA levels of genes implicated in lipid uptake, neosynthesis, β-oxidation and secretion were determined by RT-qPCR.</p

ADRP increases the HCV entry through an upregulation of occludin.

<p><b>(A)</b> Control and ADRP-overexpressing cells were transfected with the subgenomic replicon <i>in vitro</i> transcript. <b>(B)</b> Cells were transduced with HCVpp or VSV-Gpp used as control. After transfection or transduction, luciferase activity was measured by a luciferase assay. <b>(C)</b> mRNA levels of HCV receptors were measured by RT-qPCR in control and ADRP-overexpressing cells. <b>(D and E)</b> Occludin protein level in GFP or ADRP transduced cells was determined by immunoblotting. <b>(F)</b> mRNA level of occludin in GFP and PLIN5 transduced cells was determined by RT-qPCR.</p

HCV increases occludin expression level <i>via</i> a mechanism dependent on ADRP.

<p>Huh-7 were transfected with Jc1 RNA (HCV) and occludin mRNA <b>(A)</b> and protein <b>(B-C)</b> expressions were determined 48 h post transfection by RT-qPCR and immunoblotting respectively. <b>(D)</b> Control or ADRP-silenced cells were transfected or not with Jc1 full length RNA (HCV). Forty eight hours post infection, mRNA occludin level was determined by RT-qPCR. <b>(E)</b> Correlation between ADRP and occludin mRNA expression level in HCV infected patients (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146000#pone.0146000.t001" target="_blank">Table 1</a>). Spearman’s r = 0.5236.</p