Innate immune response gene expression profiles characterize primary antiphospholipid syndrome.

Primary antiphospholipid syndrome (PAPS) is a systemic autoimmune disorder characterized by thromboembolic episodes and pregnant morbidity with an increasing clinical importance. To gain insight into the pathogenesis of PAPS, we have investigated the gene expression profiles that characterize peripheral blood mononuclear cells derived from PAPS patients. We show that the transcriptional activity of genes involved in innate immune responses, such as toll-like receptor 8 and CD14, as well as downstream genes of this pathway, such as STAT1, OAS2, TNFSF13 and PLSCR1 are significantly increased in PAPS patients. In addition, the expression of monocyte-specific cytokines is also elevated in PAPS mononuclear cells stimulated in vitro with lipopolysaccharide. Taken together, these results reveal a 'response to pathogen' signature in PAPS, which could reflect an altered monocyte activity. Finally, microarray analyses also revealed a reduced expression of genes coding for proteins involved in transcriptional control. Interestingly, a significant proportion of them exhibit E2F-binding sites in their promoter, suggesting that a deregulated RB/E2F activity could play a role in the pathogenesis of antiphospholipid syndrome.

Fabrication of SU-8 multilayer microstructures based on successive CMOS compatible adhesive bonding and releasing steps.

This paper describes a novel fabrication process based on successive wafer-level bonding and releasing steps for stacking several patterned layers of the negative photoresist EPON SU-8. This work uses a polyimide film to enhance previous low temperature bonding technology. The film acts as a temporary substrate where the SU-8 is photopatterned. The poor adhesion between the polyimide film and SU-8 allows the film to be released after the bonding process, even though the film is still strong enough to carry out photolithography. Using this technique, successive adhesive bonding steps can be carried out to obtain complex 3-D multilayer structures. Interconnected channels with smooth vertical sidewalls and freestanding structures are fabricated. Unlike previous works, all the layers are photopatterned before the bonding process yielding sealed cavities and complex three-dimensional structures without using a sacrificial layer. Adding new SU-8 layers reduces the bonding quality because each additional layer decreases the thickness uniformity and increases the polymer crosslinking level. The effect of these parameters is quantified in this paper. This process guarantees compatibility with CMOS electronics and MEMS. Furthermore, the releasing step leaves the input and the output of the microchannels in contact with the outside world, avoiding the usual slow drilling process of a cover. Hence, in addition to the straightforward integration of electrodes on a chip, this fabrication method facilitates the packaging of these microfluidic devices.

Quantitative analysis of two-dimensional gel electrophoresis protein patterns: a method for studying genetic relationships among Globodera pallida populations.

A method based in two-dimensional protein gel electrophoresis has been developed in order to improve the analysis of genetic relationships among populations of Globodera. It has been used to estimate genetic divergence among nine Globodera pallida nematode populations. Sixty-one anonymous polypeptide spots were resolved using silver-stained high-resolution 2D gels and they were quantified in each population to establish genetic variation among G. pallida populations. The results of this analysis were compared with those obtained after a study of allelic frequency variation, which was carried out using seven previously described loci. Genetic distances among populations were calculated by means of both studies, the quantitative analysis and the allelic frequency variation, and phylogenetic trees were constructed for each type of analysis. A correlation analysis between the two distance matrices was carried out and a bootstrap analysis was performed to determine the strength of the clusters obtained with each method. The results obtained support the idea that quantitative protein analysis can be successfully applied to phylogenetic analysis of G. pallida populations.