Increased expression of YAP1 in prostate cancer correlates with extraprostatic extension.

Objective:Yes associated protein 1 (YAP1) is a member of the Hippo pathway, acting as a transcriptional coactivator. To elucidate the role of YAP1 and phosphorylated (p)YAP1 in prostate cancer (PCa) tumorigenesis, we investigated their expression in clinical samples of PCa and cell lines. Methods:Fifty-four tumor, adjacent nontumor, and prostate intraepithelial neoplasia (PIN) tissues from patients with PCa after radical prostatectomy were selected from a retrospective cohort and studied using immunohistochemistry (IHC). Protein and mRNA expression levels of YAP1 were evaluated by Western blot analysis and quantitative real-time reverse transcription PCR, respectively, in cancer cell lines. Publicly available gene expression datasets were downloaded to analyze YAP1 mRNA and protein levels in PCa tissue samples. Results:IHC analysis of PCa tissues revealed that YAP1 staining intensities were moderate to weak in the nucleus and cytoplasm of tumor cells, whereas adjacent normal epithelia showed strong staining. We observed that benign prostates were characterized by higher expression levels of both nuclear (P=0.004) and cytosolic (P=0.005) YAP1. pYAP1 staining was weak in the cytoplasm and absent in the nucleus of all the tissues investigated. YAP1 expression was an indicator of extraprostatic extension (EPE). The level of YAP1 was negatively correlated with the level of the androgen receptor (AR) in The Cancer Genome Atlas dataset and Western blot analysis of cell lines. Conclusions:Our study suggested that YAP1 expression is heterogeneous in PCa tissue samples; therefore, YAP1 might play different roles in different aspects of PCa progression. This might involve AR-YAP1 interplay in PCa

Interkulturelle Medien und politische Partizipation am Beispiel der MigrantInnen aus der Türkei in Wien

Diese Arbeit beinhaltet das Leben, den Lebensstil, die Hoffnung, Wünsche, Ängste, Erlebnisse usw. der Zugewanderten aus der Türkei nach Wien, wobei folgende Punkte wie: Integration, Migration, die Gesellschaft und politische Partizipation genauer unter die Lupe genommen wurden, sind im Hinblick auf den Einfluss der türkischsprachigen Printmedien in Österreich auf die politische Partizipation ausgerichtet. Um die Fragestellungen zum Thema „Politische Partizipation der MigrantInnen aus der Türkei in Österreich und den Einfluss der türkischsprachigen Medien bei der politischen Partizipation der Einwanderer“ zu bearbeiten, ist eine empirische Untersuchung durchgeführt worden. Diese Art der Untersuchung ist gewählt worden um die Ergebnisse möglichst standardisiert zu erfassen

Cloning expression and purification of durum wheat metallothionein domains and structural modelling

Metallothioneins (MTs) are small proteins with high cysteine content and high binding capacity for metals including Zn, Cu and Cd. They exist in a wide range of organisms and are classified in one super-family according to the distribution of cysteine motifs in their sequences. Type I plant MTs, similar to mammalian MTs, have the cysteine motifs clustered in the N-and C-termini constituting the β- and α-domains, respectively. In type I plant MTs the two domains are connected by a long (about 42 amino acids) hinge region whose structural and functional properties are unclear. A mt gene in Cd resistant durum wheat coding for a type I MT (dMT) was identified and the recombinant protein (dMT) was overexpressed in E. coli as GST fusion (GSTdMT) (Bilecen et al., 2005). In the present study, for detailed structural investigations; GST-fusion constructs of β-hinge, α-hinge and the isolated hinge domains of dMT were overexpressed in E. coli. Proteins were purified and were characterized by size exclusion chromatography, SDS- and native-PAGE, limited trypsinolysis, inductively coupled plasma optical emission spectroscopy (ICP-OES), UV-vis absorption spectroscopy, dynamic light scattering (DLS) and small-angle solution X-ray scattering (SAXS). GSTdMT fusion constructs were purified as stable dimeric forms in solution. Cd2+/protein ratio were found to be 1.53 ± 0.6 for GSTβ-hingedMT and 1.5 ±0.8 for GSTα-hingedMT. GSThingedMT and GSTα-hingedMT were easily cleaved from the hinge region confirming its susceptibility to proteolytic attack. Hinge region in the GSTβ-hingedMT construct, on the other hand, was protected. Cleavage was not observed in the constructs within the β- and α-domains which strongly indicated their compact structure with the bound Cd2+ ions. SAXS measurement results indicated that GSThingedMT, GSTβ-hingedMT and GSTα-hingedMT have symmetric, elongated shapes. ab initio models revealed structures with GST molecules forming electron dense regions at the center of mass and the partner dMT domains extending from this region. Evidence from Cd-binding, tryptic cleavage and SAXS models suggest that hingedMT, β-hingedMT and α-hingedMT structures fold independently of GST in the fusion constructs. The combination of SAXS results with biochemical data indicated extended structures for the dMT domains and supports the dumbbell model previously proposed for durum wheat MT (Bilecen et al., 2005)

Common and Distinctive Functions of the Hippo Effectors Taz and Yap in Skeletal Muscle Stem Cell Function

Funded by MRC grant G11001931 to HW and PSZ, European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement number 262948–2 (BIODESIGN), with additional support from AssociationFrançaise Contre les Myopathies, BBSRC, Sarcoma UK and Friends of Anchor.Peer reviewedPublisher PD

MST1 is a multifunctional caspase-independent inhibitor of androgenic signaling

The MST1 serine - threonine kinase, a component of the RASSF1-LATS tumor suppressor network, is involved in cell proliferation and apoptosis and has been implicated in cancer. However, the physiologic role of MST1 in prostate cancer (PCa) is not well understood. Here, we investigated the possibility of a biochemical and functional link between androgen receptor (AR) and MST1 signaling. We showed that MST1 forms a protein complex with AR and antagonizes AR transcriptional activity as shown by coimmunoprecipitation (co-IP), promoter reporter analysis, and molecular genetic methods. In vitro kinase and site-specific mutagenesis approaches indicate that MST1 is a potent AR kinase; however, the kinase activity of MST1 and its proapoptotic functions were shown not to be involved in inhibition of AR. MST1 was also found in AR - chromatin complexes, and enforced expression of MST1 reduced the binding of AR to a well-characterized, androgen-responsive region within the prostate-specific antigen promoter. MST1 suppressed PCa cell growth in vitro and tumor growth in mice. Because MST1 is also involved in regulating the AKT1 pathway, this kinase may be an important new link between androgenic and growth factor signaling and a novel therapeutic target in PCa. ©2011 AACR

Possibilities and limitations of capillary gas chromatography and mass spectrometry in the analysis of polychlorinated biphenyls

The possibilities and limitations of analyses of polychlorinated biphenyls (PCBs) by capillary gas chromatography and capillary gas chromatography-mass spectrometry have been investigated. Metal capillary columns (WCOT) coated with Apiezon L and OV-101 were not suitable for PCB analyses. Good results were obtained in the separation of model mixtures of PCBs and of Aroclor 1242 on a glass capillary column coated with OV-101. Sources of error are indicated that may be encountered in the characterization of PCB components in Aroclor 1242 by standard additions, Kováts' retention indices and mass spectrometry. The direct coupling of a capillary column (WCOT) to a mass spectrometer produced spectra of the main PCB components, many of which could be used in the identification of isomeric PCBs