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Platelet Aggregation Inhibitors from Philippine Marine Invertebrate Samples Screened in a New Microplate Assay

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Abstract

A new microplate assay for Ca2+-induced platelet aggregation as detected by Giemsa dye was used to screen marine invertebrate samples from the Philippines for inhibitors of human platelet aggregation. Out of 261 crude methanol extracts of marine sponges and tunicates, 25 inhibited aggregation at 2 mg/ml. Inhibition of agonist-induced aggregation in an aggregometer was used to confirm results of the microplate assay and to determine the specific mode of inhibition of 2 samples. The marine sponge Xestospongia sp. yielded a xestospongin/araguspongine-type molecule that inhibited collagen-induced aggregation by 87% at 2 µg/ml, and epinephrine-induced aggregation by 78% at 20 µg/ml, while the marine sponge Aplysina sp. yielded 5,6-dibromotryptamine, which inhibited epinephrine-induced aggregation by 51% at 20 µg/ml. In this study we have found that the microplate assay is a simple, inexpensive, yet useful preliminary tool to qualitatively screen a large number of marine samples for antiplatelet aggregation activity.

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Acknowledgements

This work was supported by the International Foundation for Science grant F1760 (G.P.C.), a UPMSI grant (G.P.C.), and collaborative projects of NCDDG, NCI, US NIH (G.P.C. and C.M.I). Marine samples were collected by P.A. Aliño and his group at UPMSI, and identified by M.K. Harper, University of Utah. The Chronolog aggregometer of the UNILAB Clinical Pathology Lab was used for the aggregometer assay. The authors thank I.T. Sandoval and F. Chung for technical support and Dr. M. Garson for her comments and suggestions on the paper.

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Authors and Affiliations

  1. Marine Science Institute, University of the Philippines, Quezon City 1101, Philippines

    Sheila Marie V. Pimentel, Zenaida P. Bojo, Amy V. D. Roberto, Jose Enrico H. Lazaro, Gina C. Mangalindan & Gisela P. Concepcion

  2. Medical Affairs Division, United Laboratories, Mandaluyong City, Philippines

    Leila M. Florentino & Pilar Lim-Navarro

  3. Department of Medicinal Chemistry, University of Utah, Salt Lake City, USA

    Deniz Tasdemir & Chris M. Ireland

Authors
  1. Sheila Marie V. Pimentel
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  2. Zenaida P. Bojo
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  3. Amy V. D. Roberto
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  4. Jose Enrico H. Lazaro
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  5. Gina C. Mangalindan
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  6. Leila M. Florentino
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  7. Pilar Lim-Navarro
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  8. Deniz Tasdemir
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  9. Chris M. Ireland
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  10. Gisela P. Concepcion
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Correspondence to Gisela P. Concepcion.

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Pimentel, S.V., Bojo, Z.P., Roberto, A.V. et al. Platelet Aggregation Inhibitors from Philippine Marine Invertebrate Samples Screened in a New Microplate Assay . Mar. Biotechnol. 5, 395–400 (2003). https://doi.org/10.1007/s10126-002-0080-3

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  • DOI: https://doi.org/10.1007/s10126-002-0080-3

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