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Purpose: The vascular endothelial growth factors (VEGF-A, -C, -D) and the VEGF receptors (VEGFR-1, -2, and -3) are important molecular markers in angiogenesis and lymphangiogenesis. This study elucidates the prognostic significance of these molecular markers in tumor cells as well as in the tumor stroma of resected non–small cell lung cancer tumors. Experimental Design: Tumor tissue samples from 335 resected patients with stage I to IIIA disease were obtained and tissue microarrays were constructed from duplicate cores of tumor cells and surrounding stromal tissue from each resected specimen. Immunohistochemistry was used to evaluate the expression of each molecular marker. Microvessel density was assessed by CD34 immunohistochemical staining. Results: In univariate analyses, high tumor cell expression of VEGF-A (P = 0.0005), VEGFR-1 (P = 0.013), VEGFR-2 (P = 0.006), and VEGFR-3 (P = 0.0003) were negative prognostic indicators for disease-specific survival (DSS). In tumor stroma, however, high expression of VEGF-A (P = 0.017), VEGF-C (P = 0.003), VEGF-D (P = 0.009), VEGFR-1 (P = 0.01), and VEGFR-2 (P = 0.019) correlated with good prognosis. There was no significant correlation between microvessel density and DSS. In multivariate analyses, high expression in tumor cells of VEGFR-3 (P = 0.007) was an independent negative prognostic factor for DSS, whereas in stromal cells, high VEGF-C (P = 0.004) expression had an independent positive survival impact. Conclusion: These are the first tissue microarray data in non–small cell lung cancers showing a positive prognostic impact by highly expressed angiogenic markers in tumor stroma, with VEGF-C as a major independent prognostic indicator.

Background: The Atlantic cod (Gadus morhua) is a groundfish of great economic value in fisheries and an emerging species in aquaculture. Genetic markers are needed to identify wild stocks in order to ensure sustainable management, and for marker-assisted selection and pedigree determination in aquaculture. Here, we report on the development and evaluation of a large number of Single Nucleotide Polymorphism (SNP) markers from the alignment of Expressed Sequence Tag (EST) sequences in Atlantic cod. We also present basic population parameters of the SNPs in samples of North-East Arctic cod and Norwegian coastal cod obtained from three different localities, and test for SNPs that may have been targeted by natural selection.

Protein kinase D (PKD), a family of serine/threonine kinases, can be activated by a multitude of stimuli in a protein kinase C-dependent or -independent manner. PKD is involved in signal transduction pathways controlling cell proliferation, apoptosis, motility, and protein trafficking. Despite its versatile functions, few genuine in vivo substrates for PKD have been identified. In this study we demonstrate that the transcription factor cAMPresponse element-binding protein (CREB) is a direct substrate for PKD. PKD1 and CREB interact in cells, and activated PKD1 provokes CREB phosphorylation at Ser-133 both in vitro and in vivo. A constitutive active mutant of PKD1 stimulates GAL4-CREB-mediated transcription in a Ser-133-dependent manner, activates CRE-responsive promoters, and increases the expression of CREB target genes. PKD1 also enhances transcription mediated by two other members of the CREB family, ATF-1 and CREM. Our results describe a novel mechanism for PKD-induced signaling through activation of the transcription factor CREB and suggest that stimulus-induced phosphorylation of CREB, reported to be mediated by protein kinase C, may involve downstream activated PKD.The mammalian PKD 3 family of serine/threonine kinases includes the isoforms PKD1 (mouse PKD and human PKC), PKD2, and PKD3 (also named PKC). PKD was originally considered to be a member of the PKC family (1, 2) but is now classified in the calcium/calmodulin-dependent kinase group based on sequence similarities in the kinase domain (3). The PKDs share a similar architecture consisting of a C-terminal catalytic domain, an N-terminal regulatory domain that encompasses two cysteine-rich regions (C1a and C1b), and a pleckstrin homology (PH) domain (4 -7). A comparison of the amino acid sequences of PKD1-3 reveals that the highest homology lies in the catalytic domain, followed by C1a, C1b, and PH domain, suggesting that isoform-specific functions may be due to the regulatory part of the kinase (8).PKD1 can be activated by several mechanisms. The most studied mechanism involves a sequential activation of phospholipase C that results in the generation of the second messengers inositol 1,4,5-triphosphate and diacylglycerol (DAG) and subsequent activation of the classical (␣, ␤I, ␤II, and ␥) and novel (␦, ⑀, , and ) PKC isoforms. Binding of DAG to the C1b domain of PKD1 directs its translocation to the plasma membrane where activated novel PKC phosphorylates PKD1 at Ser-744/748 in the activation loop, causing activation of the enzyme. The activated enzyme can be imported via its C1b motif into the nucleus, where it transiently accumulates before being exported to the cytosol through a CRM1-dependent nuclear export pathway that requires the PH domain of PKD (4 -7). Mutations and deletions in the regulatory domain induce activation of PKD to various extents, and the entire regulatory domain has an inhibitory effect on the kinase activity (9). Indeed, PKD can also be activated through caspase-mediated cleavage of the regulatory domain (10). G␤␥ subunits ...