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Toxicology
 
March 21 - 23
Paulo Oliveira, João Laranjinha e Leonor Almeida
International Courses on Toxicology 2007
Center for Neurosciences and Cell Biology
University of Coimbra, Portugal
Auditório da Reitoria, March 21-23, 2007
“Stem Cells as a Tool in Toxicology”

Organized by the Cell and Molecular Toxicology Research Area
INFORMATIONS: Paulo J. Oliveira (pauloliv@ci.uc.pt)
The course is the 2007 edition of the “International Courses on Toxicology”, organized by the Cell and Molecular Toxicology Group of the CNC on a yearly basis. The 2007 edition will be held at the Auditório da Reitoria of the University of Coimbra.
Objectives: The objective of this course is to provide theoretical information on the use of stem cells as a tool in toxicology. Stem cells are an important new tool for the development of unique in vitro model systems to test drugs and chemicals and also a potential to predict or anticipate toxicity in humans.

Invited Speakers
  1. Ernest Arenas (Karolinska Institute, Sweden)
  2. Sérgio Dias (Instituto Português de Oncologia, Portugal)
  3. Horst Spielmann (Federal Institute for Risk Assessment, Berlin, Germany)
  4. Edward Perkins (University of Minnesota-Duluth, USA
Opening Lecture
João Ramalho-Santos (Center for Neurosciences and Cell Biology, Coimbra, Portugal)

LECTURERS BIO

Ernest Arenas (Karolinska Institute, Sweden)
Full Professor of Stem Cell Neurobiology
Present Address:         
Laboratory of Molecular Neurobiology,
Department of Medical Biochemistry and Biophysics,
Karolinska Institute, BZ Lab., Scheeleväg, 1
Retzius building A1
17 177 Stockholm - Sweden
E-mail: ernest@ ki.se
Selected publications:
Castelo-Branco G, Sousa KM, Bryja V, Pinto L, Wagner J, Arenas E. Ventral midbrain glia express region-specific transcription factors and regulate dopaminergic neurogenesis through Wnt-5a secretion. Mol Cell Neurosci (2006) 31:251-262.
Kele J, Simplicio N, Ferri AL, Mira H, Guillemot F, Arenas E, Ang SL. Neurogenin 2 is required for the development of ventral midbrain dopaminergic neurons. Development (2006) 133:495-505.
Rawal N, Castelo-Branco G, Sousa KM, Kele J, Kobayashi K, Okano H, Arenas E. Dynamic temporal and cell type-specific expression of Wnt signaling components in the developing midbrain. Exp Cell Res (2006) 312:1626-1636.
Parish CL, Parisi S, Persico MG, Arenas E, Minchiotti G. Cripto as a target for improving embryonic stem cell-based therapy in Parkinson´s disease. Stem Cells (2005) 23:471-476.

Edward Perkins (University of Minnesota-Duluth, USA)
Assistant Professor, Biochemistry and Molecular Biology
UMD Biochem/Molecular Biology
249 Medical School, D602
1035 University Dr
Duluth, MN 55812 - USA
e-mail: eperkins@d.umn.edu
Selected publications
Holy, J., Lamont, G., and Perkins, E. (2006) Disruption of nucleocytoplasmic trafficking of cyclin D1 and topoisomerase II by sanguinarine. BMC Cell Biol 7, 13
Lindenbaum, M., Perkins, E., Csonka, E., Fleming, E., Garcia, L., Greene, A., Gung, L., Hadlaczky, G., Lee, E., Leung, J., MacDonald, N., Maxwell, A., Mills, K., Monteith, D., Perez, C.F., Shellard, J., Stewart, S., Stodola, T., Vandenborre, D., Vanderbyl, S., Ledebur, H.C. Jr. A mammalian artificial chromosome engineering system (ACE System) applicable to biopharmaceutical protein production, transgenesis and gene-based cell therapy. Nucleic Acids Res. 2004 Dec 7;32(21):e172. 
Vanderbyl, S., MacDonald, G. N., Sidhu, S., Gung, L., Telenius, A., Perez, C., and Perkins, E. (2004) Transfer and stable transgene expression of a mammalian artificial chromosome into bone marrow-derived human mesenchymal stem cells. Stem Cells 22, 324-333
Stewart, S., MacDonald, N., Perkins, E., DeJong, G., Perez, C., Lindenbaum, M. (2002) Retrofitting of a satellite repeat DNA-based murine artificial chromosome (ACes) to contain loxP recombination sites. Gene Ther. 9(11):719-23. 
Sérgio Dias (Instituto Português de Oncologia, Portugal)
Researcher, Group Leader
CIPM, Instituto Portugues de Oncologia Francisco Gentil, Lisboa - Portugal
e-mail: sergidias@ipolisboa.min-saude.pt.

Selected publications:
Fragoso, R., Pereira, T., Wu, Y., Zhu, Z., Cabeçadas, J., Dias, S. (2006) VEGFR-1 (FLT-1) activation modulates acute lymphoblastic leukemia localization and survival within the bone marrow, determining the onset of extramedullary disease. Blood. 107(4):1608-16.
Dias, S., Silva, H. Jr., Cumano, A., Vieira, P. (2005) Interleukin-7 is necessary to maintain the B cell potential in common lymphoid progenitors. J Exp Med. 2005;201(6):971-9.
Avecilla, S. T., Hattori, K., Heissig, B., Tejada, R., Liao, F., Shido, K., Jin, D. K., Dias, S., Zhang, F., Hartman, T. E., Hackett, N. R., Crystal, R. G., Witte, L., Hicklin, D. J., Bohlen, P., Eaton, D., Lyden, D., de Sauvage, F., and Rafii, S. (2004) Chemokine-mediated interaction of hematopoietic progenitors with the bone marrow vascular niche is required for thrombopoiesis. Nat Med 10, 64-71
Hattori, K., Heissig, B., Wu, Y., Dias, S., Tejada, R., Ferris, B., Hicklin, D. J., Zhu, Z., Bohlen, P., Witte, L., Hendrikx, J., Hackett, N. R., Crystal, R. G., Moore, M. A., Werb, Z., Lyden, D., and Rafii, S. (2002) Placental growth factor reconstitutes hematopoiesis by recruiting VEGFR1(+) stem cells from bone-marrow microenvironment. Nat Med 8, 841-849

 Horst Spielmann (Federal Institute for Risk Assessment, Berlin, Germany)
Director
Department 3 - Scientific Service
Federal Institute for Risk Assessment
Diedersdorfer Weg 1, 12277 Berlin - Germany
e-mail: Horst.Spielmann@bfr.bund.de
Selected references
Seiler, A.E., Buesen, R., Visan, A., Spielmann, H. (2006) Use of murine embryonic stem cells in embryotoxicity assays: the embryonic stem cell test. Methods Mol Biol. 329:371-95.
Bremer, S., Cortvrindt, R., Daston, G., Eletti, B., Mantovani, A., Maranghi, F., Pelkonen, O., Ruhdel, I., Spielmann, H. (2005) Reproductive and developmental toxicity. Altern Lab Anim. 33 Suppl 1:183-209.
Spielmann, H. (2005) Predicting the risk of developmental toxicity from in vitro assays.
Toxicol Appl Pharmacol. 207(2 Suppl):375-80.
Genschow, E., Spielmann, H., Scholz, G., Pohl, I., Seiler, A., Clemann, N., Bremer, S., Becker, K. (2004) Validation of the embryonic stem cell test in the international ECVAM validation study on three in vitro embryotoxicity tests. Altern Lab Anim. 32(3):209-44.

João Ramalho-Santos (Center for Neurosciences and Cell Biology, Coimbra, Portugal)
Associate Professor, Department of Zoology, University of Coimbra
Center for Neurosciences and Cell Biology
Department of Zoology, University of Coimbra
e-mail: jramalho@ci.uc.pt
Selected publications:
St John, J.C., Amaral, A., Bowles, E., Oliveira, J.F., Lloyd, R., Freitas, M., Gray, H.L., Navara, C.S., Oliveira, G., Schatten, G.P., Spikings, E., Ramalho-Santos, J. (2006) The analysis of mitochondria and mitochondrial DNA in human embryonic stem cells. Methods Mol Biol;331:347-74.
Lloyd, R.E., Lee, J.H., Alberio, R., Bowles, E.J., Ramalho-Santos, J., Campbell, K.H., St John, J.C. (2006) Aberrant nucleo-cytoplasmic cross-talk results in donor cell mtDNA persistence in cloned embryos. Genetics. 172(4):2515-27.
St John, J.C., Ramalho-Santos, J., Gray, H.L., Petrosko, P., Rawe, V.Y., Navara, C.S., Simerly, C.R., Schatten, G.P. (2005) The expression of mitochondrial DNA transcription factors during early cardiomyocyte in vitro differentiation from human embryonic stem cells. Cloning Stem Cells.;7(3):141-53.
Thompson, W.E., Ramalho-Santos, J., Sutovsky, P. (2003) Ubiquitination of prohibitin in mammalian sperm mitochondria: possible roles in the regulation of mitochondrial inheritance and sperm quality control. Biol Reprod. ;69(1):254-60.
 
Schedule
Day I – Wednesday, March 21
10:00-11:00 – Opening lecture: João Ramalho-Santos Portuguese Ph.D. students followed by general discussion). Moderators: Sérgio Dias, Ernest Arenas Interesting artifacts: The many lives and times of embryonic stem cells
11:00-12:00 – Sérgio Dias - Stem and progenitor cells in health and disease
12:00-12:15 - Coffee break
12:15-13:15 – Sérgio Dias - Endothelial progenitor cells in angiogenesis
14:45-17:00 – Afternoon presentations I (includes five short presentations from
Day II – Thursday, March 22
09:00-10:00 – Horst Spielmann - The use of stem cells in toxicologychromosomes for stem cell transfection
11:00-11:15 - Coffee break
11:15-12:15 – Ernest Arenas - Stem cells as tools for neuroprotection, cellreplacement and toxicology
14:30-17:00 – Afternoon presentations II (includes four presentations of Ph.D. students from the CNC doctoral programme). Moderators: Edward Perkins, Horst Spielmann

10:00-11:00 – Edward Perkins - Design and synthesis of mammalian artificial
 Day III – Friday, March 23
09:00-10:00 – Horst Spielmann - Improvement of the embryonic stem cell testby establishing molecular endpoints of tissue-specificdevelopment
10:00-11:00 – Ernest Arenas - Regulation of dopaminergic neuron development.by Wnts
11:15-12:15 - Coffee break
11:30-12:30 – Edward Perkins - Chromosome-based gene expression platforms for stem cell engineering
16:00 – 17:00 –Closing seminar: Ernest Arenas - Stem cells and Parkinson´sdisease

AFTERNOON SESSIONS
SHORT COMMUNICATIONS FROM Ph.D. STUDENTS – March 21st, 2007
1 - Co-culture models: understanding and regulation of cell proliferation and differentiation mechanisms,
Rogério P. Pirraco1,2, AP Marques1,2, RL Reis1,2
(1) 3B´s Research Group – Biomaterials, Biodegradables and Biomimetics, Dept. of Polymer Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal.
(2) IBB – Institute for Biotechnology and Bioengineering, PT Government Associated Laboratory, Braga, Portugal.
2 – Scale-up of mouse embryonic stem cell expansion in a stirred culture system.
Ana Margarida Fernandes1, Tiago Fernandes1, Margarida Diogo1, Cláudia Lobato da Silva1, Domingos Henrique2, Joaquim MS Cabral1
(1) - Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Lisboa, Portugal
(2) - Instituto de Medicina Molecular, Faculdade de Medicina, Lisboa, Portugal
3 - Fat tissue: A natural reserve of Stem cells - isolation methods and osteogenic differentiation of Adipose Stem Cells (ASC).
Tommaso Rada1,2, M.E. Gomes1,2, R.L. Reis1,2
(1) 3B´s Research Group – Biomaterials, Biodegradables and Biomimetics, Dept. of Polymer Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal.
(2) IBB – Institute for Biotechnology and Bioengineering, PT Government Associated Laboratory, Braga, Portugal.
4 - Role of nitric oxide produced by microglia on the proliferation of neural stem cells from the subventricular zone
Bruno P. Carreira1, Gabriel Costa1, Aida Costa1, Ângela Inácio1, Fabienne Agasse1, Denis Soulet, António F. Ambrósio1,3, Caetana M. Carvalho1, Inês M. Araújo1.
(1) Center for Neuroscience and Cell Biology, Department of Zoology, University of Coimbra, Coimbra, Portugal
(2) Wallenberg Neuroscience Center, Neuronal Survival Unit, Lund University, Lund, Sweden
(3) Center of Ophthalmology of Coimbra, IBILI, Faculty of Medicine, University of Coimbra, 3000-354 Coimbra, Portugal.
5 - Role of Growth Factors and Extracellular Matrix on the Differentiation of
Neural Stem/Precursor Cells
Ana Silva1, J. Pereira2, J. Bettencourt-Relvas2, C. R. Oliveira1 , A. C. Rego1.
(1) Center for Neuroscience and Cell Biology and Institute of Biochemistry, Faculty of Medicine, University of Coimbra, Portugal,
(2) Institute of Cell Biology, Swiss Federal Institute of Technology, Zurich, Switzerland
6 - Rat retinal cells and toxicity: role of neuropeptide Y
Ana Rita Álvaro1,2, Inês Araújo1, João Martins1,2, Joana Rosmaninho-Salgado1, Ana Tellechea1, António F. Ambrósio1,2, Cláudia Cavadas1,3
(1) Center for Neuroscience and Cell Biology, University of Coimbra, Portugal
(2) Center of Ophthalmology of Coimbra, IBILI, Faculty of Medicine, University of Coimbra, Portugal
(3) Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal
7 - Neurogenesis in the intact and damaged adult brain: focus on inflammation
Liliana Bernardino1, Agasse F. 1, Grade S. 1, Ferreira R. 1, Silva B.A. 1, Malva J.O.1
(1) Center for Neuroscience and Cell Biology, University of Coimbra, Portugal
(to be confirmed)
10 minutes presentations followed by questions from the moderator and audience

BEB STUDENTS PRESENTATIONS – March 22st, 2007
1) Evaluation of the haematotoxicity of Taxol and Adriamycin in frozen human umbilical blood chord cells – protection by antioxidants
Ana Burgeiro, Alexandra Rosa, Vera Ribeiro
2) Functional activity of mitochondria in cultured neural precursor cells – role of inhibitors and pro-oxidant agents
Sofia Grade, Ana Soares, Joana Gaspar
3) Alterations in mitochondrial generation of free radicals, antioxidant enzymes and mass during the spontaneous differentiation of human embryonic stem cells
Sara Sousa, Pedro Branco, Marina Brito
4) Generation of hepatocyte-line from human embryonic stem cells and comparison of cell toxicity of ethanol, bile acids and mitochondrial poisons between the two cell states.Gil Costa, Daniel Nunes, Joana Neves

O objectivo da tarefa que vos propomos consiste em escrever uma proposta para financiamento de um determinado projecto de investigação. Supondo que vocês queriam obter dinheiro de uma das “muitas” entidades que existem em Portugal para o efeito, como o fariam?
A proposta será apresentada oralmente, em 15 minutos máximo (Inglês, claro). Não se esqueçam das coisas chave: hipótese e/ou objectivos, introdução, interesse para a ciência/prática clínica e tarefas específicas.
Estarei ao vosso dispor.
Boa sorte
 
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