Einfluss von ZNS-Verletzung auf die m-RNA-Expression der Protease-Aktivierenden Rezeptoren (PARs) und Indentifizierung eines Interaktionspartners für PAR-2 in Retina

Protease-activated receptor, optic nerve crush, focal ischemia, protein-protein interaction, alpha crystallin A

Protease-activated receptor (PAR)-1 and PAR-2 protect rat astrocytes from apoptotic cell death via differentially regulating JNK isoform-specific release of the chemokine GRO/CINC-1 : two novel protective pathways in brain

Protease-activated receptor; c-Jun N-terminal kinase (JNK); thrombin; neuroprotection; siRNA

Analysis of two newly identified protease-activated receptor-2-interacting proteins, Jab1 and p24A, and their role in receptor signalling

Protease-activated receptor, receptor signalling, receptor trafficking, receptor resensitization, protein interaction

Synergistic inflammatory signaling in airway epithelial cells : control of expression levels of protease activated receptors and interleukin-8 release

Protease-activated receptor, interleukin, thrombin, trypsin, asthma

Protease-activated receptor 2 and [alpha]-crystallin : interactions and functional implications

Magdeburg, Univ., Fak. für Naturwiss., Diss., 2010

Regulation der "Secretory Leukocyte Protease Inhibitor"-Expression in Magenepithelzellen unter dem Einfluss der Infektion mit Helicobacter pylori

Magdeburg, Univ., Med. Fak., Diss., 2011

Distribution of the human intracellular serpin protease inhibitor 8 in human tissues.

Ovalbumin-like serine protease inhibitors are mainly localized intracellularly and their in vivo functions are largely unknown. To elucidate their physiological role(s), we studied the expression of one of these inhibitors, protease inhibitor 8 (PI-8), in normal human tissues by immunohistochemistry using a PI-8-specific monoclonal antibody. PI-8 was strongly expressed in the nuclei of squamous epithelium of mouth, pharynx, esophagus, and epidermis, and by the epithelial layer of skin appendages, particularly by more differentiated epithelial cells. PI-8 was also expressed by monocytes and by neuroendocrine cells in the pituitary gland, pancreas, and digestive tract. Monocytes showed nuclear and cytoplasmic localization of PI-8, whereas neuroendocrine cells showed only cytoplasmic staining. In vitro nuclear localization of PI-8 was confirmed by confocal analysis using serpin-transfected HeLa cells. Furthermore, mutation of the P(1) residue did not affect the subcellular distribution pattern of PI-8, indicating that its nuclear localization is independent of the interaction with its target protease. We conclude that PI-8 has a unique distribution pattern in human tissues compared to the distribution patterns of other intracellular serpins. Additional studies must be performed to elucidate its physiological role.

Flippase (FLP) recombinase-mediated marker recycling in the dermatophyte Arthroderma vanbreuseghemii.

Biological processes can be elucidated by investigating complex networks of relevant factors and genes. However, this is not possible in species for which dominant selectable markers for genetic studies are unavailable. To overcome the limitation in selectable markers for the dermatophyte Arthroderma vanbreuseghemii (anamorph: Trichophyton mentagrophytes), we adapted the flippase (FLP) recombinase-recombination target (FRT) site-specific recombination system from the yeast Saccharomyces cerevisiae as a selectable marker recycling system for this fungus. Taking into account practical applicability, we designed FLP/FRT modules carrying two FRT sequences as well as the flp gene adapted to the pathogenic yeast Candida albicans (caflp) or a synthetic codon-optimized flp (avflp) gene with neomycin resistance (nptII) cassette for one-step marker excision. Both flp genes were under control of the Trichophyton rubrum copper-repressible promoter (PCTR4). Molecular analyses of resultant transformants showed that only the avflp-harbouring module was functional in A. vanbreuseghemii. Applying this system, we successfully produced the Ku80 recessive mutant strain devoid of any selectable markers. This strain was subsequently used as the recipient for sequential multiple disruptions of secreted metalloprotease (fungalysin) (MEP) or serine protease (SUB) genes, producing mutant strains with double MEP or triple SUB gene deletions. These results confirmed the feasibility of this system for broad-scale genetic manipulation of dermatophytes, advancing our understanding of functions and networks of individual genes in these fungi.

Expression of protease-activated receptors in arthritic synovial tissues

Clinical and experimental evidence suggests that synovial thrombin formation in arthritic joints is prominent and deleterious, leading to exacerbation of rheumatoid arthritis (RA). In this context, cellular effects of thrombin mediated by the protease-activated receptors (PARs) in arthritic joints may be of paramount significance. Four PARs have now been identified. PAR1, PAR3, and PAR4 can all be activated by thrombin whereas PAR2 is activated by trypsin and few other proteases.We first explored PARs expression in RA synovial tissues. Synovial membranes from 11 RA patients were analyzed for PARs expression by RT-PCR and by immunohistology. PAR4 was found in all the biopsies, whereas the expression of PAR1, PAR 2 and PAR3 was more restricted (8/11, 5/11 and 3/11 respectively). In the arthritic synovial membrane of murine antigen-induced arthritis (AIA) we found coexpression of the four different PARs. Next, we explored the functional importance of PAR1 during AIA in vivo using PAR-1 deficient mice. The phenotype of PAR1-deficient mice (n = 22), based on the analysis of arthritis severity (as measured by 99 m tecnetium uptake, histological scoring and intra-articular fibrin measurements) was similar to that of wild-type mice (n = 24). In addition, the in vivo production of antibodies against mBSA was also similar. By contrast, the mBSA-induced in vitro lymph node cell proliferation was significantly decreased in PAR1-deficient mice as compared with controls. Accordingly, mBSA-induced production of interferon-γ by lymph node cells in culture was significantly decreased in PAR1-deficient mice as compared with controls, whereas opposite results were observed for production of IL-10.

A peptide inhibitor of C-jun N-terminal kinase modulates hepatic damage and the inflammatory response after hemorrhagic shock and resuscitation

Hemorrhage and resuscitation (H/R) leads to phosphorylation of mitogen-activated stress kinases, an event that is associated with organ damage. Recently, a specific, cell-penetrating, protease-resistant inhibitory peptide of the mitogen-activated protein kinase c-JUN N-terminal kinase (JNK) was developed (D-JNKI-1). Here, using this peptide, we tested if inhibition of JNK protects against organ damage after H/R. Male Sprague-Dawley rats were treated with D-JNKI-1 (11 mg/kg, i.p.) or vehicle. Thirty minutes later, rats were hemorrhaged for 1 h to a MAP of 30 to 35 mmHg and then resuscitated with 60% of the shed blood and twice the shed blood volume as Ringer lactate. Tissues were harvested 2 h later. ANOVA with Tukey post hoc analysis or Kruskal-Wallis ANOVA on ranks, P < 0.05, was considered significant. c-JUN N-terminal kinase inhibition decreased serum alanine aminotransferase activity as a marker of liver injury by 70%, serum creatine kinase activity by 67%, and serum lactate dehydrogenase activity by 60% as compared with vehicle treatment. The histological tissue damage observed was blunted after D-JNKI-1 pretreatment both for necrotic and apoptotic cell death. Hepatic leukocyte infiltration and serum IL-6 levels were largely diminished after D-JNKI-1 pretreatment. The extent of oxidative stress as evaluated by immunohistochemical detection of 4-hydroxynonenal was largely abrogated after JNK inhibition. After JNK inhibition, activation of cJUN after H/R was also reduced. Hemorrhage and resuscitation induces a systemic inflammatory response and leads to end-organ damage. These changes are mediated, at least in part, by JNK. Therefore, JNK inhibition deserves further evaluation as a potential treatment option in patients after resuscitated blood loss.

La responsabilidad social corporativa en las empresas del IBEX-35

La responsabilitat social corporativa és un concepte molt genèric i, en moltes ocasions, bastant abstracte i desconegut que no significa que no estigui molt present i citat avui en dia. El treball de la responsabilitat social corporativa en les empreses de l’IBEX-35 té per objectiu ensenyar i educar al lector sobre què consisteix exactament aquest concepte i conèixer més profundament les raons per les quals és imprescindible incorporar-lo en el món empresarial, sota un context de globalització i de competència permanent. Tanmateix, una vegada interioritzat aquesta matèria és molt important per als clients, proveïdors i membres afectats per les activitats de l’empresa rebre la informació de forma detallada sobre quina metodologia, accions i missió que pren l’empresa per aconseguir complir amb els reptes i objectius que se li plantegen i es marca sobre RSC cosa que fa necessari per a assolir una bona conducta, coordinació i feed-back positiu entre l’empresa i els diferents grups d’interès una bona comunicació amb aquests grups d’interès. Això es possible gràcies a l’elaboració d’un informe de RSC. Per dur-lo a terme amb coherència, claredat i, sobretot, perquè pugui arribar a ser útil per a la gestió i govern de l’organització, serà necessari definir-lo i emmarcar-lo en els àmbits i temàtiques que té que contenir i el procediment per elaborar-lo. Tot informe precisa d’anàlisis per a avaluar la qualitat de la informació reflexada i, d’aquesta manera, poder realitzar accions correctores oportunes. Mitjançant la definició d’una sèrie d’indicadors de desenvolupament i altres, intentarem aproximar-nos a l’anàlisi dels continguts de les memòries de les empreses que componen l’IBEX-35 de l’any 2006, ja que podem considerar-les com les més representatives i més influents d’Espanya. Amb tot això sabrem quina és la situació en l’àmbit general dels informes que presenten aquestes empreses i podrem recomanar per a que les empreses puguin completar-lo i respondre més eficientment les necessitats dels futurs lectors d’aquest informe i obtenir millor imatge corporativa.

Mutations of the Serine Protease CAP1/Prss8 Lead to Reduced Embryonic Viability, Skin Defects, and Decreased ENaC Activity.

CAP1/Prss8 is a membrane-bound serine protease involved in the regulation of several different effectors, such as the epithelial sodium channel ENaC, the protease-activated receptor PAR2, the tight junction proteins, and the profilaggrin polypeptide. Recently, the V170D and the G54-P57 deletion mutations within the CAP1/Prss8 gene, identified in mouse frizzy (fr) and rat hairless (fr(CR)) animals, respectively, have been proposed to be responsible for their skin phenotypes. In the present study, we analyzed those mutations, revealing a change in the protein structure, a modification of the glycosylation state, and an overall reduction in the activation of ENaC of the two mutant proteins. In vivo analyses demonstrated that both fr and fr(CR) mutant animals present analogous reduction of embryonic viability, similar histologic aberrations at the level of the skin, and a significant decrease in the activity of ENaC in the distal colon compared with their control littermates. Hairless rats additionally had dehydration defects in skin and intestine and significant reduction in the body weight. In conclusion, we provided molecular and functional evidence that CAP1/Prss8 mutations are accountable for the defects in fr and fr(CR) animals, and we furthermore demonstrate a decreased function of the CAP1/Prss8 mutant proteins. Therefore, fr and fr(CR) animals are suitable models to investigate the consequences of CAP1/Prss8 action on its target proteins in the whole organism.