Search

To elucidate the role of gene candidates involved in pulmonary hypertension (PH) associated with systemic sclerosis (SSc).Gene candidates were identified through microarray experiments performed on Affymetrix GeneChip Human Exon 1.0 ST arrays in endothelial progenitor cell (EPC)-derived endothelial cells (ECs) obtained from patients with SSc-associated PH, patients with SSc without PH, and healthy control subjects. Expression of identified gene candidates was assessed by quantitative sandwich enzyme-linked immunosorbent assay in the serum, and by immunohistochemistry in lesional lung tissue. The functional importance of the identified gene candidates was then evaluated in fos-related antigen 2-transgenic (Fra-2-Tg) mice that spontaneously develop SSc-like features associated with an intense pulmonary vascular remodeling.Microarray experiments revealed that the matrix metalloproteinase 10 (MMP-10) gene was the top up-regulated gene in SSc-associated PH EPC-derived ECs. Circulating serum proMMP10 concentrations were markedly increased in patients with SSc-associated PH compared to SSc patients without PH and healthy controls. Consistent with these observations, a strong MMP10 staining of the thickened wall of distal pulmonary arteries was found both in the lungs of patients with SSc-associated PH and in the lungs of Fra-2-Tg mice. Daily treatment of Fra-2-Tg mice with neutralizing anti-MMP10 antibodies did not significantly affect the development and severity of pulmonary fibrosis, but did reverse established PH and markedly reduced pulmonary vascular remodeling by reducing cell proliferation, cell survival, and the platelet-derived growth factor signaling axis.Gene expression profiling of EPC-derived ECs identified MMP10 as a novel candidate gene in SSc-associated PH. MMP10 is overexpressed in the serum and pulmonary arteries of patients with SSc-associated PH, and its blockade alleviates PH in the Fra-2-Tg mouse model. MMP10 appears to be a prospective treatment target for this devastating disorder.

Treatment for fibrosis represents a critical unmet need, because fibrosis is the leading cause of death in industrialized countries, and there is no effective therapy to counteract the fibrotic process. The development of fibrosis relates to the interplay between vessel injury, immune cell activation, and fibroblast stimulation, which can occur in various tissues. Immunotherapies have provided a breakthrough in the treatment of immune diseases. The glycoprotein OX40-OX40 ligand (OX40L) axis offers the advantage of a targeted approach to costimulatory signals with limited impact on the whole immune response. Using systemic sclerosis (SSc) as a prototypic disease, we report compelling evidence that blockade of OX40L is a promising strategy for the treatment of inflammation-driven fibrosis. OX40L is overexpressed in the fibrotic skin and serum of patients with SSc, particularly in patients with diffuse cutaneous forms. Soluble OX40L was identified as a promising serum biomarker to predict the worsening of lung and skin fibrosis, highlighting the role of this pathway in fibrosis. In vivo, OX40L blockade prevents inflammation-driven skin, lung, and vessel fibrosis and induces the regression of established dermal fibrosis in different complementary mouse models. OX40L exerts potent profibrotic effects by promoting the infiltration of inflammatory cells into lesional tissues and therefore the release of proinflammatory mediators, thereafter leading to fibroblast activation.

Background Systemic sclerosis (SSc) is a severe fibrotic disease with no effective treatment. The OX40-OX40L axis is emerging as a new potential target in autoimmune diseases offering the advantage of a targeted approach on co-stimulatory signals with limited impact on the whole immune response and very preliminary data suggested its involvement in SSc. Objectives To investigate the contribution of OX40L in SSc Methods We assessed the expression of OX40L in fibrotic skin and in the serum of SSc-patients. The efficacy of a targeted therapy against OX40L in vivo was evaluated using complementary experimental mouse models. Results Expression of OX40L protein was higher in fibrotic skin of SSc-patients as compared to healthy controls (p=0.003), particularly in diffuse cutaneous forms. Soluble OX40 levels were significantly higher in SSc-patients (n=177) than in healthy controls (n=100). Longitudinal analyses of our cohort showed that high level of serum OX40L at baseline was highly predictive of worsening of dermal and lung fibrosis during the follow-up (HR: 8.28 [2.11–32.50] and 4.60 [1.52–12.18], P Invalidation of OX40L prevented dermal fibrosis in the bleomycin mouse model with a decrease in dermal thickness, hydroxyprolin content and myofibroblasts count. OX40L invalidation was associated with a decrease in T cell, B cell and macrophage infiltrates and in cytokine release (IL-6, TNF-α) in lesional skin of mice. A transcriptomic approach identified two main pathways regulated by OX40L blockade: NF kappa B and AP 1 signaling. Then pharmacologic approaches, using a monoclonal anti-OX40L antibody, demonstrated that the blockade of OX40L not only prevented dermal fibrosis, but also induced regression of established fibrosis. Thereafter, the use of this monoclonal antibody demonstrated in the fra-2 transgenic mouse model marked protection from fibrosing alveolitis (measured by CT-scan (Figure) and histology) and vessel remodeling leading to pulmonary arterial hypertension. Conclusions Altogether, our results show that OX40L is as an attractive target in inflammation-driven fibrosis. OX40L appears as a very promising biomarker in SSc and an appealing potential therapy to reduce both skin and lung fibrosis. Disclosure of Interest None declared

Primary care interventions directed at drinking and alcohol problems today constitute a highly important issue for service development, training and research. WHO has clone much to encourage work in this area, and in this journal feature we publish a summary by Thomas Babor, Marcus Grant et al. of a WHO multi-national clinical trial on the treatment of alcohol problems in the primary care setting. This summary is followed by comments from, Canada, Australia, Scotland, Norway, Denmark and Sweden and a reply from THomas Babor. We are grateful to the WHO investigators and the expert group of commentators, and to Robert West who took responsibility for organizing this series of contributions

Ambient ozone has been linked to the worsening of symptoms of patients with obstructive diseases such as chronic obstructive pulmonary disease (COPD) and asthma. We investigated the role of cathepsin S on ozone-induced airway hyperresponsiveness (AHR) and inflammation, using the selective cathepsin S inhibitor, Compound A. Balb/c mice were exposed to ozone at a concentration of 3 ppm or air for 3 h, following administration by gavage of Compound A or vehicle. Bronchoalveolar lavage (BAL) was performed 3 h and 20-24 h following exposure, AHR was measured at 20-24 h only. Ozone exposure, compared to air exposure increased BAL cathepsin S levels, AHR and BAL inflammatory cells. Compound A (30 mg kg(-1) p.o.) dosing compared to vehicle dosing inhibited ozone-induced AHR (-logPC100 vehicle: -0.70+/-0.12, n=8 vs. cathepsin S inhibitor: -1.30+/-0.06, P

Acute chest syndrome (ACS), the most common cause of death among adults with sickle cell disease and a significant threat to children, is often triggered by bone marrow embolization, especially among adults. We have recently developed a sickle transgenic mouse model for acute chest that consists of injecting a mixture of bone marrow and mouse fat into the tail vein of a sickle transgenic partial knockout mouse. The mouse used was a NY1 partial KO expressing 100% human α and 55% βS globins. Mice were injected with bone marrow (BM) from two femurs (about 15 mg) and purified peritoneal fat (about 15 mg, as murine BM lacks fat), and examined at 0, 6, 24, and 48 hours post-injection. Creatine kinase (CK), predominantly the MM form, increased above baseline levels at 6, 24, and 48 hours in NY1 partial KO mice; whereas, in control C57Bl mice, there was an initial increase at 6 hours with a return to baseline values at 24 and 48 hours. sVCAM levels were not significantly different between C57Bl and NY1 partial KO mice at baseline: 717±203 in C57Bl mice and 752±147 in NY1 partial KO mice (N=15 and 11, respectively); after injection of fat/BM sVCAM levels nearly doubled in NY1 partial KO mice and remained elevated at 48 hours. In fat/BM injected C57Bl mice, sVCAM levels remained essentially constant from 0 to 48 hours. Mouse lungs were harvested at 48 hours after saline perfusion and inflated with 10% phosphate buffered formalin. The hematoxylin and eosin stained lung sections were normal among the C57Bl control mice examined 48 hours after fat/BM injection, but among NY1 partial KO mice edema, inflammation, and congestion were found with numerous red cells entrapped, despite saline perfusion. In summary, this preparation shows promise as an animal model of ACS. Pulmonary edema and elevated levels of sVCAM are characteristic features of ACS in sickle cell patients and are found, after fat/BM injection, in our sickle mice but not in control mice. The mice survived the procedure, indicating that it will be possible to study the natural course of lung injury and interventions that may ameliorate injury.

Theoretical incompressible velocity distributions for three airfoils in various cascade arrangements and for different entering-air angles are presented. Design charts and design formulas relating cascade angle, entering-air angle, turning angle, and blade camber are presented for 10 per cent thick airfoils at solidity 1. An empirical turning-angle formula is developed and is found to be reasonably accurate and rather generally applicable.

Electrical and magnetic properties have been measured in the tantalum‐carbon system from Ta2C to TaC1.0. A large variation of these properties has been found within the wide homogeneity range of TaC: the resistivity increases with carbon deficiency from 25 to about 150 μΩ. cm, and the magnetic susceptibility changes its sign, the material being paramagnetic at stoichiometry and diamagnetic at the low‐carbon phase boundary. The temperature coefficient of the resistivity has also been found to be characteristic for each composition and to depend strongly on the Ta:C ratio.A method for preparation of homogeneous samples of a predetermined carbon content has been developed. The experimental results are discussed in the light of previous theories.