General characteristics of cases and controls.

<p>General characteristics of cases and controls.</p

Etiology of the mitral regurgitation in the 54 control patients.

<p>Etiology of the mitral regurgitation in the 54 control patients.</p

Supplemental material for Riociguat treatment for portopulmonary hypertension: a subgroup analysis from the PATENT-1/-2 studies

<p>Supplemental material for Riociguat treatment for portopulmonary hypertension: a subgroup analysis from the PATENT-1/-2 studies by Rodrigo Cartin-Ceba, Michael Halank, Hossein-Ardeschir Ghofrani, Marc Humbert, John Mattson, Arno Fritsch and Michael Krowka in Pulmonary Circulation</p

Temporal Asthma Patterns Using Repeated Questionnaires over 13 Years in a Large French Cohort of Women

<div><p>Variable expression is one aspect of the heterogeneity of asthma. We aimed to define a variable pattern, which is relevant in general health epidemiological cohorts. Our objectives were to assess whether: 1) asthma patterns defined using simple asthma questions through repeated measurements could reflect disease variability 2) these patterns may further be classified according to asthma severity/control. Among 70,428 French women, we used seven questionnaires (1992–2005) and a comprehensive reimbursement database (2004–2009) to define three reliable asthma patterns based on repeated positive answers to the ever asthma attack question: “never asthma” (n = 64,061); “inconsistent” (“yes” followed by “no”, n = 3,514); “consistent” (fully consistent positive answers, n = 2,853). The “Inconsistent” pattern was related to both long-term (childhood-onset asthma with remission in adulthood) and short-term (reported asthma attack in the last 12 months, associated with asthma medication) asthma variability, showing that repeated questions are relevant markers of the variable expression of asthma. Furthermore, in this pattern, the number of positive responses (1992–2005) predicted asthma drug consumption in subsequent years, a marker of disease severity. The “Inconsistent” pattern is a phenotype that may capture the variable expression of asthma. Repeated answers, even to a simple question, are too often neglected.</p></div

Figure 5

<p>Mouse lung PECAM-1 immunostaining (<b>A</b>). (<b>B</b>) Number of vessels per 100 alveoli. (<b>C</b>) Mouse lung F4/80 immunostaining. (<b>D</b>) Number of macrophages per 100 alveoli. Values are mean±SEM. *<i>P</i><0.02 compared with wild-type mice exposed to normoxia and ^σ<i>P</i><0.02 compared with wild-type mice exposed to hypoxia.</p

Key Role of the Endothelial TGF-β/ALK1/Endoglin Signaling Pathway in Humans and Rodents Pulmonary Hypertension

<div><p>Mutations affecting transforming growth factor-beta (TGF-β) superfamily receptors, activin receptor-like kinase (ALK)-1, and endoglin (ENG) occur in patients with pulmonary arterial hypertension (PAH). To determine whether the TGF-β/ALK1/ENG pathway was involved in PAH, we investigated pulmonary TGF-β, ALK1, ALK5, and ENG expressions in human lung tissue and cultured pulmonary-artery smooth-muscle-cells (PA-SMCs) and pulmonary endothelial cells (PECs) from 14 patients with idiopathic PAH (iPAH) and 15 controls. Seeing that ENG was highly expressed in PEC, we assessed the effects of TGF-β on Smad1/5/8 and Smad2/3 activation and on growth factor production by the cells. Finally, we studied the consequence of ENG deficiency on the chronic hypoxic-PH development by measuring right ventricular (RV) systolic pressure (RVSP), RV hypertrophy, and pulmonary arteriolar remodeling in ENG-deficient (<i>Eng^+/−</i>) and wild-type (<i>Eng^+/+</i>) mice. We also evaluated the pulmonary blood vessel density, macrophage infiltration, and cytokine expression in the lungs of the animals. Compared to controls, iPAH patients had higher serum and pulmonary TGF-β levels and increased ALK1 and ENG expressions in lung tissue, predominantly in PECs. Incubation of the cells with TGF-β led to Smad1/5/8 phosphorylation and to a production of FGF2, PDGFb and endothelin-inducing PA-SMC growth. Endoglin deficiency protected mice from hypoxic PH. As compared to wild-type, <i>Eng^+/−</i> mice had a lower pulmonary vessel density, and no change in macrophage infiltration after exposure to chronic hypoxia despite the higher pulmonary expressions of interleukin-6 and monocyte chemoattractant protein-1. The TGF-β/ALK1/ENG signaling pathway plays a key role in iPAH and experimental hypoxic PH via a direct effect on PECs leading to production of growth factors and inflammatory cytokines involved in the pathogenesis of PAH.</p></div

Figure 2

<p>Effect of increasing TGF-β doses on Smad 1,5,8 and Smad 2,3 phosphorylation, respectively, in pulmonary endothelial cells (PECs) from controls and from patients with idiopathic pulmonary hypertension (iPAH) (<b>A</b> and <b>B</b>). Protein levels were normalized for β-actin. Values are means±SEM normalized for results without TGF-β. *<i>P</i><0.05 compared to relevant controls without TGF-β, ^§<i>P</i><0.05 compared to control PECs under the same conditions. (<b>C</b>) Growth of pulmonary-artery smooth-muscle cells (PA-SMCs) from controls in response to serum-free media derived from cultured PECs from controls and stimulated by TGF-β with or without anti-ENG antibody (ENG Ab).Values are mean±SEM. *<i>P</i><0.05 compared to basal condition, ^§<i>P</i><0.05 compared to PA-SMCs stimulated with PEC medium.</p

Figure 5

<p>Mouse lung PECAM-1 immunostaining (<b>A</b>). (<b>B</b>) Number of vessels per 100 alveoli. (<b>C</b>) Mouse lung F4/80 immunostaining. (<b>D</b>) Number of macrophages per 100 alveoli. Values are mean±SEM. *<i>P</i><0.02 compared with wild-type mice exposed to normoxia and ^σ<i>P</i><0.02 compared with wild-type mice exposed to hypoxia.</p

The open bars indicate the results in controls and the closed bars in patients with idiopathic pulmonary hypertension (iPAH).

<p>TGF-β was assayed using an ELISA in serum and lung homogenates (controls, n = 15; iPAH patients, n = 14) (<b>A</b> and <b>B</b>). (<b>C</b>) TGF-β mRNA measured in pulmonary endothelial cells (PECs) and pulmonary-artery smooth-muscle cells (PA-SMCs; controls, n = 7; iPAH patients, n = 7). (<b>D</b>) TGF-β receptor expression: <i>ALK1, ALK5</i>, and <i>ENG</i> mRNA measured in PECs and PA-SMCs (controls, n = 7; iPAH patients, n = 7). (<b>E</b>): ENG, ALK1, and ALK5 protein expression in LUNG, PECs and PA-SMCs. Protein levels were normalized for β-actin (controls, n = 7; iPAH patients, n = 7). Values are mean±SEM. *<i>P</i><0.05 and **<i>P</i><0.01 compared with controls.</p

Effect of pulmonary endothelial cells (PECs) from controls incubated with TGF-β with or without anti-ENG antibody (ENG Ab) on mRNA expression of A) <i>preproET-1</i>, (B) <i>PDGFa</i>, (C) <i>PDGFb</i>, (D) <i>FGF2</i>, (E) <i>EGF</i>, (F) <i>MCP-1</i>, and (G) <i>IL-6</i>.

<p>Values are mean±SEM.*<i>P</i><0.05 compared to PECs without TGF-β treatment.</p