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We investigated whether human peripheral blood mononuclear cells (PBMC) express prothrombinase following stimulation with bacterial lipopolysaccharide (LPS). LPS-stimulated PBMC devoid of contaminating platelets failed to activate prothrombin directly. Addition of platelets did not result in expression of prothrombinase in the absence of factor X whereas the combination of platelets and factor X resulted in strong prothrombinase activity on LPS-activated cells. The induced prothrombinase was dependent on tissue factor, as the activity was completely inhibited by an anti-tissue factor antibody. Our data suggest that platelet/monocyte cooperation is important in the generation of prothrombinase activity in response to endotoxin.

Extensive expression of craniofacial related homeobox genes in canine mammary sarcomas Journal Breast Cancer Research and Treatment Publisher Springer Netherlands ISSN 0167-6806 (Print) 1573-7217 (Online) Issue Volume 118, Number 2 / November, 2009 Category Preclinical Study DOI 10.1007/s10549-008-0243-7 Pages 333-343 Subject Collection Medicine SpringerLink Date Tuesday, December 02, 2008 Preclinical Study Extensive expression of craniofacial related homeobox genes in canine mammary sarcomas Helena Wensman1 , Hanna Göransson2, Karl-Johan Leuchowius3, Sara Strömberg3, Fredrik Pontén3, Anders Isaksson2, Gerard Roel Rutteman4, Nils-Erik Heldin3, Gunnar Pejler1 and Eva Hellmén1 (1) Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, P.O.Box 7011, 750 07 Uppsala, Sweden (2) Department of Medical Sciences, Uppsala University, Uppsala, Sweden (3) Department of Genetics and Pathology, Uppsala University, Uppsala, Sweden (4) Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands Received: 10 September 2008 Accepted: 30 October 2008 Published online: 2 December 2008 Abstract The global gene expression in three types of canine mammary tumors: carcinoma, fibrosarcoma and osteosarcoma were investigated by Affymetrix gene array technology. Unsupervised clustering analysis revealed a close clustering of the respective tumor types, with fibrosarcomas clustering close to the osteosarcomas and the carcinomas clustering closer to non-malignant mammary tissues (NMTs). A number of epithelial markers were expressed in both carcinomas and NMTs, whereas the sarcomas expressed genes related to mesenchymal differentiation. A comparison of the gene expression profile of the sarcomas versus carcinoma/NMTs revealed that the sarcomas, in particular the osteosarcomas, showed a striking upregulation of a panel of homeobox genes previously linked to craniofacial bone formation. In line with this finding, osteosarcomas showed an upregulation of bone morphogenetic proteins (BMPs) and of genes associated with retinoic acid signaling. Increased homeobox gene expression in sarcomas was also confirmed at the protein level by immunohistochemical analysis of tumor tissue, and in an osteosarcoma cell line after stimulation by BMP-2. These findings suggest that the development of mammary sarcomas specifically involves triggering of a set of homeobox genes related to neural crest and craniofacial bone development. Electronic supplementary material The online version of this article (doi:10.1007/s10549-008-0243-7) contains supplementary material, which is available to authorized users. Keywords Canine mammary tumors - Mammary sarcoma - Mammary osteosarcoma - Homeobox transcription factor - Gene expression profiling - Craniofacial

Purpose/Objective: It becomes increasinglyclear that mast cells (MCs) are not only key players in allergic diseases (e.g.asthma), but seem to play a complex role in neuroinflammatory diseases such as multiple sclerosis and stroke. However, their role during and after mechanical CNS trauma is not clear. In the present study, we have investigated the effectsof MC-deficiency on the histological and clinical outcome after spinal cordhemisection at thoraic level T8 resulting in a complete transection of the dorsomedial and ventral corticospinal tract. Functional recovery in SCI mice was tested with the Basso Mouse Scale. Spinal cord sections were analyzed by immunofluorescence. RTPCR and Western blotting were used to analyze cytokine/chemokine mRNA and protein levels. In degradation assays murine recombinant IL-1b, IL-4, IL-6, IL-10, IL-13, TNF-a and MCP-1 were incubated with supernatant from BMCMC derived from either C57BL/6 or from mMCP4 -/-mice. Cleaved fragments were identified using tris-tricine SDS-PAGE and analyzed by intensity analysis. Results: We show that MC-deficient kitW-sh/W-sh mice display significantly increased astrogliosis and T cell infiltration as well as significantly reduced clinical outcome after SCI compared to wildtype mice. In addition, MC-deficient mice show significantly increased levels of MCP-1, TNF-a, IL-10 and IL-13 protein levels in the spinal cord after SCI. Mice deficient in mMCP-4, a MC-specific chymase, also showed increased MCP-1, IL-6 and IL-13 protein levels in spinal cord samples and a decreased functional outcome after SCI. A degradation assay using supernatant from MCs derived from either mMCP4 -/- or wildtype mice revealed that mMCP-4 cleaves MCP-1, IL-6 and IL-13, suggesting a protective role for MC proteases in neuroinflammation. These data indicate that MCs may be protective after SCI and that they may reduce CNS inflammation by degrading inflammation-associated cytokines via the mast cell-specific chymase mMCP-4. Conclusions: In summary, our results suggest a new and complex mechanism how MCs and their proteases may protect the CNS from exacerbated and/or chronic inflammation after damage. Publication appears at doi: 10.1111/imm.12002