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Domenico Ribatti - One of the best experts on this subject based on the ideXlab platform.

  • tryptase and chymase are angiogenic in vivo in the Chorioallantoic Membrane Assay
    The International Journal of Developmental Biology, 2011
    Co-Authors: Domenico Ribatti, Beatrice Nico, Girolamo Ranieri, Vincenzo Benagiano, Enrico Crivellato
    Abstract:

    Human mast cells (MCs) are divided in two types depending on the expression of tryptase and chymase in their granules. Literature data indicate that both tryptase and chymase are angiogenic, but there is currently no evidence of their direct angiogenic activity in vivo. In this study, we have investigated the capacity of tryptase and chymase to promote vasoproliferation in chick embryo Chorioallantoic Membrane (CAM), a well established in vivo Assay to study angiogenesis and anti-angiogenesis. The results showed that both tryptase and chymase stimulate angiogenesis and that the response is similar to that obtained with vascular endothelial growth factor (VEGF), a well-known angiogenic cytokine, and confirm the angiogenic activity of these two proteases stored in MC granules.

  • vascular endothelial growth factors synthesized by human lung mast cells exert angiogenic effects
    The Journal of Allergy and Clinical Immunology, 2009
    Co-Authors: Aikaterini Detoraki, Nella Prevete, Rosaria I Staiano, Giorgio Giannattasio, Francescopaolo Granata, Arturo Genovese, Amato De Paulis, Massimo Triggiani, Domenico Ribatti, Gianni Marone
    Abstract:

    Background Angiogenesis and lymphangiogenesis are critical for several allergic, inflammatory, and neoplastic disorders. Mast cells infiltrate the sites of inflammation and tumors. Objective We sought to characterize the expression and functions of vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) in human mast cells. Methods VEGF expression was evaluated by means of RT-PCR and Western blotting in primary human lung mast cells and in the mast cell lines LAD-2 and HMC-1. Angiogenic activity of mast cell supernatants was determined by using the chick embryo Chorioallantoic Membrane Assay. VEGFR expression was assessed by means of RT-PCR and flow cytometry. Modified Boyden chambers were used for chemotaxis Assay. Results Human mast cells express VEGF-A, VEGF-B, VEGF-C, and VEGF-D at both the mRNA and protein level. Prostaglandin E 2 (PGE 2 ) enhanced the expression of VEGFA , VEGFB , and VEGFC , whereas an adenosine analog (5′-[N-ethylcarboxamido] adenosine [NECA]) increased VEGFA , VEGFC , and VEGFD expression. In addition, PGE 2 and NECA enhanced VEGF-A release, and supernatants of PGE 2 - and NECA-activated human lung mast cells induced angiogenic responses in the Chorioallantoic Membrane Assay that were inhibited by an anti-VEGF-A antibody. Mast cells expressed mRNA for VEGFR1 and VEGFR2 . These receptors were present on the mast cell surface. VEGF-A 165 , VEGF-B 167 , VEGF-C, VEGF-D, and placental growth factor 1 induced mast cell chemotaxis. These chemotactic effects were mediated by the activation of both VEGFR-1 and VEGFR-2. Conclusion Our data indicate that human mast cells are both a source and a target of angiogenic and lymphangiogenic factors and therefore might play a role in inflammatory and neoplastic angiogenesis through the expression of several forms of VEGFs and their receptors.

  • in vivo silencing of aquaporin 1 by rna interference inhibits angiogenesis in the chick embryo Chorioallantoic Membrane Assay
    Cellular and Molecular Biology, 2006
    Co-Authors: Giulia Maria Camerino, Domenico Ribatti, Grazia Paola Nicchia, Maria Maddalena Dinardo, Maria Svelto, Antonio Frigeri
    Abstract:

    : Aquaporin-1 (AQP1) is a water channel protein mainly expressed in endothelial and epithelial cells of many tissues, including the vasculature where it serves to increase cell Membrane water permeability. Previous studies in active multiple myeloma patients and in AQP1 KO mice indicated an involvement of AQP1 in physiological and tumor angiogenesis. To understand the physiological role of AQP1 in angiogenesis, we used a 21-nucleotide small interfering RNA duplexes (siRNA) to knockdown AQP1 in the chick embryo Chorioallantoic Membrane (CAM), a commonly used in vivo Assay to study both angiogenic and angiostatic molecules. Chicken AQP1 sequence was identified and utilized to synthesize a siRNA directed to the AQP1 sequence. We then tested the efficiency of the siRNA in vitro, using an AQP1 transfected cell line. The level of AQP1 protein reduction obtained using siRNA was 98 % and 92 % after 1 and 2 day transfection respectively. RNA interference experiments were then performed in vivo by using the CAM Assay. Results showed that after 4 days of treatment, AQP1 siRNA was able to strongly inhibit angiogenesis. This is the first study showing the in vivo use of RNA interference technique in the CAM Assay. Our results strongly support the hypothesis that AQP1 could have a key role in physiological and pathological angiogenesis.

  • the gelatin sponge Chorioallantoic Membrane Assay
    Nature Protocols, 2006
    Co-Authors: Domenico Ribatti, Beatrice Nico, Angelo Vacca, Marco Presta
    Abstract:

    Here we present a method for the quantification of angiogenesis and antiangiogenesis in the chick embryo Chorioallantoic Membrane (CAM) based on the implantation of a gelatin sponge on the top of the growing CAM on day 8 of development. After implantation, the sponge is treated with a stimulator of blood vessel formation in the absence or presence of an angiogenesis inhibitor. On day 12, blood vessels that are growing into the sponge are counted at macroscopic and microscopic levels. The estimated timeline for carrying out this protocol is 10 d. The presence of a vascular network in the CAM requires a careful analysis to distinguish new capillaries from pre-existing ones. This limitation does not occur in the avascular cornea Assay, which may also take advantage of different genetic backgrounds when carried out in transgenic or knockout mice. Nevertheless, the gelatin sponge–CAM Assay is simple, inexpensive and suitable for large-scale screening. *Note: In the version of the article initially published online, references 6 and 7 were incorrect. The correct references are: 6. Serbedzija, G.N., Flynn, E. & Willet, C.E. Zebrafish angiogenesis: a new model for drug screening. Angiogenesis 3, 519–528 (2000). 7. Ribatti, D., Vacca, A., Roncali, L. & Dammacco, F. The chick embryo Chorioallantoic Membrane as a model for in vivo research on angiogenesis. Int. J. Dev. Biol. 40, 1189–1897 (1996). The error has been corrected in all versions of the article.

  • the first evidence of the tumor induced angiogenesis in vivo by using the Chorioallantoic Membrane Assay dated 1913
    Leukemia, 2004
    Co-Authors: Domenico Ribatti
    Abstract:

    The first evidence of the tumor-induced angiogenesis in vivo by using the Chorioallantoic Membrane Assay dated 1913

Jen Hwey Chiu - One of the best experts on this subject based on the ideXlab platform.

  • chrysin suppresses il 6 induced angiogenesis via down regulation of jak1 stat3 and vegf an in vitro and in ovo approach
    Journal of Agricultural and Food Chemistry, 2010
    Co-Authors: Kou-gi Shyu, Bao Wei Wang, Hang Chang, Yenhsu Chen, Jen Hwey Chiu
    Abstract:

    Chrysin, 5,7-dihydroxyflavone, possesses many biologic properties. This study aimed to investigate the effects and molecular mechanisms of chrysin on IL-6-induced angiogenesis in vitro and in ovo. Chicken Chorioallantoic Membrane Assay, an in ovo angiogenesis Assay, showed chrysin significantly suppressed IL-6-induced neovascularization. Furthermore, chrysin significantly suppressed human umbilical vein endothelial cell (HUVECs) migration and tube formation. The signaling pathway involved in chrysin-related antiangiogenesis was also investigated. The data indicated that chrysin is able to down-regulate the expression of glycoprotein 130 (gp130), soluble IL-6 receptor (IL-6R), phosphorylated JAK1 and STAT3, and VEGF in HUVECs. The IL-6-induced binding of STAT3 was significantly suppressed by chrysin. Moreover, chrysin did not further suppress VEGF expression with STAT3 knocked down. Taken together, the results show that chrysin suppresses IL-6-induced angiogenesis through modulation of the sIL-6R/gp130/JAK...

  • chrysin suppresses il 6 induced angiogenesis via down regulation of jak1 stat3 and vegf an in vitro and in ovo approach
    Journal of Agricultural and Food Chemistry, 2010
    Co-Authors: Chiu Mei Lin, Bao Wei Wang, Hang Chang, Yenhsu Chen, Kou-gi Shyu, Jen Hwey Chiu
    Abstract:

    Chrysin, 5,7-dihydroxyflavone, possesses many biologic properties. This study aimed to investigate the effects and molecular mechanisms of chrysin on IL-6-induced angiogenesis in vitro and in ovo. Chicken Chorioallantoic Membrane Assay, an in ovo angiogenesis Assay, showed chrysin significantly suppressed IL-6-induced neovascularization. Furthermore, chrysin significantly suppressed human umbilical vein endothelial cell (HUVECs) migration and tube formation. The signaling pathway involved in chrysin-related antiangiogenesis was also investigated. The data indicated that chrysin is able to down-regulate the expression of glycoprotein 130 (gp130), soluble IL-6 receptor (IL-6R), phosphorylated JAK1 and STAT3, and VEGF in HUVECs. The IL-6-induced binding of STAT3 was significantly suppressed by chrysin. Moreover, chrysin did not further suppress VEGF expression with STAT3 knocked down. Taken together, the results show that chrysin suppresses IL-6-induced angiogenesis through modulation of the sIL-6R/gp130/JAK1/STAT3/VEGF signaling pathway. Chrysin may provide new therapeutic potential for IL-6-induced pathological angiogenesis.

Kou-gi Shyu - One of the best experts on this subject based on the ideXlab platform.

  • chrysin suppresses il 6 induced angiogenesis via down regulation of jak1 stat3 and vegf an in vitro and in ovo approach
    Journal of Agricultural and Food Chemistry, 2010
    Co-Authors: Kou-gi Shyu, Bao Wei Wang, Hang Chang, Yenhsu Chen, Jen Hwey Chiu
    Abstract:

    Chrysin, 5,7-dihydroxyflavone, possesses many biologic properties. This study aimed to investigate the effects and molecular mechanisms of chrysin on IL-6-induced angiogenesis in vitro and in ovo. Chicken Chorioallantoic Membrane Assay, an in ovo angiogenesis Assay, showed chrysin significantly suppressed IL-6-induced neovascularization. Furthermore, chrysin significantly suppressed human umbilical vein endothelial cell (HUVECs) migration and tube formation. The signaling pathway involved in chrysin-related antiangiogenesis was also investigated. The data indicated that chrysin is able to down-regulate the expression of glycoprotein 130 (gp130), soluble IL-6 receptor (IL-6R), phosphorylated JAK1 and STAT3, and VEGF in HUVECs. The IL-6-induced binding of STAT3 was significantly suppressed by chrysin. Moreover, chrysin did not further suppress VEGF expression with STAT3 knocked down. Taken together, the results show that chrysin suppresses IL-6-induced angiogenesis through modulation of the sIL-6R/gp130/JAK...

  • chrysin suppresses il 6 induced angiogenesis via down regulation of jak1 stat3 and vegf an in vitro and in ovo approach
    Journal of Agricultural and Food Chemistry, 2010
    Co-Authors: Chiu Mei Lin, Bao Wei Wang, Hang Chang, Yenhsu Chen, Kou-gi Shyu, Jen Hwey Chiu
    Abstract:

    Chrysin, 5,7-dihydroxyflavone, possesses many biologic properties. This study aimed to investigate the effects and molecular mechanisms of chrysin on IL-6-induced angiogenesis in vitro and in ovo. Chicken Chorioallantoic Membrane Assay, an in ovo angiogenesis Assay, showed chrysin significantly suppressed IL-6-induced neovascularization. Furthermore, chrysin significantly suppressed human umbilical vein endothelial cell (HUVECs) migration and tube formation. The signaling pathway involved in chrysin-related antiangiogenesis was also investigated. The data indicated that chrysin is able to down-regulate the expression of glycoprotein 130 (gp130), soluble IL-6 receptor (IL-6R), phosphorylated JAK1 and STAT3, and VEGF in HUVECs. The IL-6-induced binding of STAT3 was significantly suppressed by chrysin. Moreover, chrysin did not further suppress VEGF expression with STAT3 knocked down. Taken together, the results show that chrysin suppresses IL-6-induced angiogenesis through modulation of the sIL-6R/gp130/JAK1/STAT3/VEGF signaling pathway. Chrysin may provide new therapeutic potential for IL-6-induced pathological angiogenesis.

Marco Presta - One of the best experts on this subject based on the ideXlab platform.

  • the gelatin sponge Chorioallantoic Membrane Assay
    Nature Protocols, 2006
    Co-Authors: Domenico Ribatti, Beatrice Nico, Angelo Vacca, Marco Presta
    Abstract:

    Here we present a method for the quantification of angiogenesis and antiangiogenesis in the chick embryo Chorioallantoic Membrane (CAM) based on the implantation of a gelatin sponge on the top of the growing CAM on day 8 of development. After implantation, the sponge is treated with a stimulator of blood vessel formation in the absence or presence of an angiogenesis inhibitor. On day 12, blood vessels that are growing into the sponge are counted at macroscopic and microscopic levels. The estimated timeline for carrying out this protocol is 10 d. The presence of a vascular network in the CAM requires a careful analysis to distinguish new capillaries from pre-existing ones. This limitation does not occur in the avascular cornea Assay, which may also take advantage of different genetic backgrounds when carried out in transgenic or knockout mice. Nevertheless, the gelatin sponge–CAM Assay is simple, inexpensive and suitable for large-scale screening. *Note: In the version of the article initially published online, references 6 and 7 were incorrect. The correct references are: 6. Serbedzija, G.N., Flynn, E. & Willet, C.E. Zebrafish angiogenesis: a new model for drug screening. Angiogenesis 3, 519–528 (2000). 7. Ribatti, D., Vacca, A., Roncali, L. & Dammacco, F. The chick embryo Chorioallantoic Membrane as a model for in vivo research on angiogenesis. Int. J. Dev. Biol. 40, 1189–1897 (1996). The error has been corrected in all versions of the article.

  • new model for the study of angiogenesis and antiangiogenesis in the chick embryo Chorioallantoic Membrane the gelatin sponge Chorioallantoic Membrane Assay
    Journal of Vascular Research, 1997
    Co-Authors: Domenico Ribatti, Angelo Vacca, Luisa Roncali, Anna Gualandris, Maria Bastaki, Monica Iurlaro, Marco Presta
    Abstract:

    Several methods for the in vivo study of angiogenesis are available, and each angiogenic Assay presents distinct advantages and disadvantages. In this study, we present a new method for the quantitati

Yenhsu Chen - One of the best experts on this subject based on the ideXlab platform.

  • chrysin suppresses il 6 induced angiogenesis via down regulation of jak1 stat3 and vegf an in vitro and in ovo approach
    Journal of Agricultural and Food Chemistry, 2010
    Co-Authors: Kou-gi Shyu, Bao Wei Wang, Hang Chang, Yenhsu Chen, Jen Hwey Chiu
    Abstract:

    Chrysin, 5,7-dihydroxyflavone, possesses many biologic properties. This study aimed to investigate the effects and molecular mechanisms of chrysin on IL-6-induced angiogenesis in vitro and in ovo. Chicken Chorioallantoic Membrane Assay, an in ovo angiogenesis Assay, showed chrysin significantly suppressed IL-6-induced neovascularization. Furthermore, chrysin significantly suppressed human umbilical vein endothelial cell (HUVECs) migration and tube formation. The signaling pathway involved in chrysin-related antiangiogenesis was also investigated. The data indicated that chrysin is able to down-regulate the expression of glycoprotein 130 (gp130), soluble IL-6 receptor (IL-6R), phosphorylated JAK1 and STAT3, and VEGF in HUVECs. The IL-6-induced binding of STAT3 was significantly suppressed by chrysin. Moreover, chrysin did not further suppress VEGF expression with STAT3 knocked down. Taken together, the results show that chrysin suppresses IL-6-induced angiogenesis through modulation of the sIL-6R/gp130/JAK...

  • chrysin suppresses il 6 induced angiogenesis via down regulation of jak1 stat3 and vegf an in vitro and in ovo approach
    Journal of Agricultural and Food Chemistry, 2010
    Co-Authors: Chiu Mei Lin, Bao Wei Wang, Hang Chang, Yenhsu Chen, Kou-gi Shyu, Jen Hwey Chiu
    Abstract:

    Chrysin, 5,7-dihydroxyflavone, possesses many biologic properties. This study aimed to investigate the effects and molecular mechanisms of chrysin on IL-6-induced angiogenesis in vitro and in ovo. Chicken Chorioallantoic Membrane Assay, an in ovo angiogenesis Assay, showed chrysin significantly suppressed IL-6-induced neovascularization. Furthermore, chrysin significantly suppressed human umbilical vein endothelial cell (HUVECs) migration and tube formation. The signaling pathway involved in chrysin-related antiangiogenesis was also investigated. The data indicated that chrysin is able to down-regulate the expression of glycoprotein 130 (gp130), soluble IL-6 receptor (IL-6R), phosphorylated JAK1 and STAT3, and VEGF in HUVECs. The IL-6-induced binding of STAT3 was significantly suppressed by chrysin. Moreover, chrysin did not further suppress VEGF expression with STAT3 knocked down. Taken together, the results show that chrysin suppresses IL-6-induced angiogenesis through modulation of the sIL-6R/gp130/JAK1/STAT3/VEGF signaling pathway. Chrysin may provide new therapeutic potential for IL-6-induced pathological angiogenesis.