Proteomic biomarkers for psoriasis and psoriasis arthritis.

UNLABELLED: Although several new biomarkers have been recently proposed for psoriasis (Ps) and psoriasis arthritis (PsA), nothing is known about their diagnostic sensitivity and specificity, and their routine use. We therefore searched in-depth for new biomarker candidates using a biobank with EDTA-plasma from 158 individuals, patients and healthy controls. Samples from 6 selected pairs (patients against healthy controls) were searched proteomically using a workflow of extensive and precise design that is highly comprehensive. Subsequent verification was performed using ELISA and the entire biobank. By proteomic methods, 208 altered proteins were identified. Of these, 15 biomarker candidates were selected for verification. Of these 15, 4 individual parameters and 11 combinations significantly discriminated between patient and control groups. These individual parameters were Zn-α2-glycoprotein, complement C3, polymeric immunoglobulin receptor, and plasma kallikrein. Significant discrimination was obtained by combinations of 2 or 3 parameters. One combination seemed suitable for diagnosing PsA. Moreover, several candidates desmoplakin, complement C3, polymeric immunoglobulin receptor, and cytokeratin 17, correlated with PASI in all patients. This first comprehensive proteomic study on non-depleted plasma identified several biomarker candidates that have not been described before as well as some known from previous studies. BIOLOGICAL SIGNIFICANCE: Our non-gel proteomic analysis is based on the highly comprehensive and significantly optimized chromatographic protein pre-fractionation. The method allows a biomarker search in non-depleted plasma. The subsequent verification by ELISA identifies several biomarker-candidates for the unbiased diagnosis of psoriasis and psoriasis arthritis. Four of the identified candidate markers might be used individually. Combinations of several parameters improve the diagnostic sensitivity and specificity. The still not validated candidates form a reserve for further evaluation. Moreover, mass spectrometric data uncover several biomarker-candidates which show diverse protein species of the same protein with opposing changes in the same sample.

A recessive mutation leading to vertebral ankylosis in zebrafish is associated with amino acid alterations in the homologue of the human membrane-associated guanylate kinase DLG3.

We describe the characterization of the zebrafish homologue of the human gene DLG3. The zebrafish dlg3 gene encodes a membrane-associated guanylate kinase containing a single PDZ domain. This gene was cloned using a gene-trap construct inserted in the gene's first intron. The insertion co-segregates with a viable mutation called humpback (hmp), which leads to formation of ankylotic vertebrae in adult fishes. Insertion and mutation have both been mapped to chromosome 12, in a segment which is syntenic with region p12 to q12 of human chromosome 17. The hmp mutant phenotype, however, appears to be due to two point mutations in the guanylate kinase domain rather than to the transgene insertion itself. The results of this study are discussed in the light of the possible function of the guanylate kinase domain.

Cortical rotation is required for the correct spatial expression of nr3, sia and gsc in Xenopus embryos.

Beta-catenin, a component of the wnt-signal-transduction pathway, is essential for the formation of the dorsal axis in Xenopus laevis embryos. On the dorsal side of the embryo, beta-catenin is translocated into the nuclei via a process linked to cortical rotation. When cortical rotation is blocked by UV-irradiation, nuclear beta-catenin is found in the vegetal pole of the embryo. Here we show that overexpression of beta-catenin in animal cap explants, in the absence of mesoderm induction, is sufficient to activate the expression of genes with dorsalizing activity such as siamois (sia) and nodal-related 3 (nr3) but not goosecoid (gsc). In embryos ventralized by UV-treatment, the expression of the dorsal-specific genes sia, nr3 and gsc is induced at the vegetal pole after the Mid-Blastula-Transition (MBT). While nr3 and sia expression continues in these embryos until gastrula stages, gsc transcription cannot be maintained. We propose that the spatial separation of the expression domains of genes with dorsalizing activities and the prospective mesodermal region results in the loss of dorsal structures in the embryo. The role of cortical rotation is to generate an overlap of the region with dorsal axis-forming activity, indicated by nuclear translocation of beta-catenin, and the prospective mesoderm in the marginal zone to assure the correct positioning of the Spemann organizer.

Determination of the hydraulic conductivity of Lamprothamnium by use of the pressure clamp.

By use of the pressure-clamp technique, the hydraulic conductivity of the brackish-water alga Lamprothamnium was found to be 5·10(-6) cm s(-1) bar(-1). The dimensions of the internodes are so small that it is possible, for the first time, to measure a complete volume relaxation upon clamping the turgor pressure to a preset value by a feedback control of the pressure probe. As theoretically predicted, the values of the hydraulic conductivity obtained from the initial slope of the volume relaxation induced by the pressure clamp are in agreement (within experimental error) with those obtained from the half-time of the relaxation process. The cell volume also obtained from the analysis of the volume relaxation is the osmotically effective cell volume and is therefore slightly smaller than the value obtained by taking the dimensions of the cell including the cell wall.

A new method for the determination of hydraulic conductivity and cell volume of plant cells by pressure clamp.

Internodes of Chara corallina were used for experiments in which cell turgor pressure was clamped by means of the pressure probe technique. Essentially, the procedure consisted of a combination of volume and turgor pressure relaxations. This technique permits the determination of the cell volume by nonoptical means. The values obtained are in agreement with the ones determined by optical means. Furthermore, the hydraulic conductivity (L(p)) was determined from the initial slope of the volume relaxation; the values thus obtained are in agreement with those calculated from the half-times of pressure relaxations. The determination of L(p) from volume relaxation measurements has the advantage that the cell volume, the volumetric elastic modulus of the cell wall, and the internal osmotic pressure do not have to be known. Furthermore, the half-time of volume relaxation is longer than that of pressure relaxation, as shown by theory and experiment. This may be used to enhance the resolution of the relaxation measurement and, thus, to improve the accuracy of L(p) determinations for higher plant cells which exhibit a very fast pressure relaxation.