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Objective. SSc is a disease characterized by inflammation and fibrosis. Heme Oxygenase-1 (HO-1) is a haem-degrading enzyme that mediates resolution of inflammation and is induced upon mediators abundantly present in SSc. We aimed to assess whether HO-1 expression/function is disturbed in SSc patients and could therefore be contributing to the ongoing inflammation. Methods. In total, 92 SSc patients and 48 healthy controls were included. By measuring total bilirubin in plasma in vivo, HO-activity was assessed. HO-1 expression levels were determined with western blot in monocytes before and after induction of HO-1 with cobalt protoporphyrin (CoPP) with or without CXCL4. Monocyte-derived dendritic cells (DCs) were stimulated with several Toll-like receptor (TLR) ligands with or without pre-stimulation with CoPP for 24 h. Cytokine levels were measured in the supernatants using the Luminex Bead Array. Results. SSc patients have lower plasma levels of bilirubin, suggestive of an aberrant HO-1 function. We demonstrated low HO-1 expression in immune cells from SSc patients, whereas induction with CoPP was able to restore HO-1 levels in DCs from SSc patients, almost normalizing the increased TLR response observed in SSc. Co-exposure to CXCL4 completely abrogated CoPP-induced HO-1 expression, suggesting that the high CXCL4 levels present in SSc patients block the normal induction of HO-1 and its function. Conclusion. We demonstrate that HO activity in SSc patients is decreased and show its functional consequences. Since CXCL4 blocks the induction of HO-1 expression, neutralization of CXCL4 in SSc patients could have clinical benefits by diminishing overactivation of immune cells and other anti-inflammatory effects of HO-1. [ABSTRACT FROM AUTHOR]

To gain insight into the different extraction processes used in laboratories in various European countries and the effect these methods might have on the number of Meloidogyne second-stage juveniles (J2) extracted from soil samples, a ring test with 2 elements was conducted. The first element used the Baermann funnel as a method performance test. The second element was a proficiency test, in which the laboratory's standard technique was compared with the Baermann method. Chilled samples of 100 cm³ (10 per extraction method) were sent to participants in insulated containers. In the nematode counts distinction was made between Meloidogyne J2, other plant-parasitic nematodes and/or saprophytes. A total number of 18 participants took part in the method performance test and 13 institutes took part in the proficiency test using 11 different extraction techniques. The initial population density in the soil was 2025 J2 (100 cm³ soil)-1, determined by Oostenbrink elutriator and 4 weeks incubation. In the method performance test the institutes found Meloidogyne J2 numbers varied between 0 and 705 J2 (100 cm³)-1. Using the Baermann extraction method the median number per institute ranged from 0 to 377 J2 (100 cm³)-1. In the proficiency test comparison amongst the different methods showed an increase of more than 100-fold, with the highest median of 3733 J2 (100 cm³)-1 and the lowest of 34 J2 (100 cm³)-1; the difference was caused mainly by the incubation effect. For the other plant-parasitic nematodes and the saprophytes, the differences between the most and least efficient extraction methods were much smaller. The variance of the Meloidogyne counts was the highest for the standard Baermann extraction technique; other similar techniques also had high variances. The automated zonal centrifuge had the lowest variance. The Baermann method is not advisable for survey purposes. [ABSTRACT FROM AUTHOR]