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Jaime Freitas completed his PhD at Wageningen University and Research (The Netherlands) and the University of California-Davis (USA) on the development of assays to increase the insight into the mechanisms of toxic interference with the thyroid hormone (TH) receptors and provide a basis for the high-throughput screening of TH-like drugs. While finishing his PhD thesis, Jaime Freitas joined the EU-funded project NANOFOL at the IBMC (Portugal), with the aim of developing a nanodevice to target activated immune cells in Rheumatoid Arthritis. Due to his growing interest in the molecular mechanisms of RA, Jaime Freitas joined the Gene Regulation group at the IBMC (Portugal) and later moved to the University of Rutgers (USA), to learn how to use state-of-the-art bioinformatic tools to elucidate the role of transcription kinetics in essential RNA-processing mechanisms using custom sequencing methods. Currently, Jaime Freitas is a Junior Researcher in the Microenvironment in New Therapies group at i3S (Portugal) working in the translational-oriented R2Bone project, specifically, in non-coding RNA-based therapies for bone regeneration and inflammation in RA.
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Identification

Personal identification

Full name
Jaime Freitas

Citation names

  • Freitas, Jaime

Author identifiers

Ciência ID
1413-D5C3-05B3
ORCID iD
0000-0003-3229-3293

Email addresses

  • jaime.freitas@i3s.up.pt (Professional)
  • freitas.jaime@gmail.com (Personal)

Telephones

Fax
  • 226099157 (Professional)
Mobile phone
  • 919149257 (Personal)
Telephone
  • 226074900 (Professional)

Addresses

  • I3S-Instituto de Investigação e Inovação em Saúde, Rua Alfredo Allen 208, lab 105S1, 4200-135, Porto, Porto, Portugal (Professional)

Websites

Knowledge fields

  • Natural sciences - Biological Sciences

Languages

Language Speaking Reading Writing Listening Peer-review
Portuguese Advanced (C1) Advanced (C1) Advanced (C1) Advanced (C1)
English Advanced (C1) Advanced (C1) Advanced (C1) Advanced (C1)
Spanish; Castilian Beginner (A1) Beginner (A1) Beginner (A1) Beginner (A1)
French Beginner (A1) Beginner (A1) Beginner (A1) Beginner (A1)
Education
Degree Classification
2017/10/14 - 2017/12/16
Concluded
 Clinical Research Associate Training Course (Pós-Graduação)
CTI Clinical Trial and Consulting Services Lisbon, Portugal
2016/09/28 - 2016/09/30
Concluded
 Hands-on workshop on Image Analysis for Imaging flow Cytometry (Pós-Graduação)
Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
2013/10/08 - 2013/10/11
Concluded
 Whole Exome Sequencing and RNA-seq data analysis (Pós-Graduação)
Instituto Gulbenkian de Ciência, Portugal
2012/12/17
Concluded
 doctor (PhD) (Doutoramento)
Major in Toxicology
Wageningen Universiteit, Netherlands
"Development and Validation of in vitro Bioassays for Thyroid Hormone Receptor Mediated Endocrine Disruption" (THESIS/DISSERTATION)
2009/10/05 - 2009/10/09
Concluded
 Toxicological Pathology (Curso de doutoramento (conclusão de unidades curriculares))
Universitair Medisch Centrum Utrecht, Netherlands
2009/07/19 - 2009/07/24
Concluded
 Entrepreneurial Boot Camp (Curso de doutoramento (conclusão de unidades curriculares))
University of Wisconsin-Madison School of Business, United States
2009 - 2009
Concluded
 Environmental Toxicology (Curso de doutoramento (conclusão de unidades curriculares))
Wageningen Universiteit en Research, Netherlands
2006/07/19
Concluded
 Química Aplicada - Ramo de Biotecnologia (Licenciatura)
Universidade Nova de Lisboa Faculdade de Ciências e Tecnologia, Portugal
"Polysome Profiling. An Improved Method for IL-6 Translational State Analysis" (THESIS/DISSERTATION)
Affiliation

Science

Category
Host institution 		Employer
2019/05/01 - Current Contracted Researcher (Research) Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
2016/01/01 - 2019/04/30 Postdoc (Research) Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
2014 - 2015 Postdoc (Research) Universidade do Porto Instituto de Biologia Molecular e Celular, Portugal
Universidade do Porto Instituto de Biologia Molecular e Celular, Portugal
2013 - 2014 Postdoc (Research) Universidade do Porto Instituto de Biologia Molecular e Celular, Portugal
Universidade do Porto Instituto de Biologia Molecular e Celular, Portugal
2012 - 2013 Researcher (Research) Universidade do Porto Instituto de Biologia Molecular e Celular, Portugal
Universidade do Porto Instituto de Biologia Molecular e Celular, Portugal
2006 - 2012/12/17 Contracted Researcher (Research) Wageningen Universiteit en Research, Netherlands
Wageningen Universiteit en Research, Netherlands
2005 - 2006 Research Trainee (Research) Universidade do Porto Instituto de Ciências Biomédicas Abel Salazar, Portugal
Universidade do Porto Instituto de Ciências Biomédicas Abel Salazar, Portugal
Projects

Grant

Designation Funders
2016 - 2019 FOLSMART - Folate-target nanodevices to activated macrophages for rheumatoid arthritis.
H2020-NMP-PILOTS-2015
Post-doc Fellow
Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
 EU Framework Programme for Research and Innovation Industrial Leadership
Concluded
2014 - 2015 Molecular mechanisms of transcription termination between closely spaced genes.
PTDC/BIA-BCM/119795/2010
Post-doc Fellow
Universidade do Porto Instituto de Biologia Molecular e Celular, Portugal
 Fundação para a Ciência e a Tecnologia
Concluded
2013 - 2014 NANOFOL - Folate-based nanobiodevices for integrated diagnosis/therapy targeting chronic inflammatory diseases.
FP7-NMP-2008-LARGE-2
Post-doc Fellow
Universidade do Porto Instituto de Biologia Molecular e Celular, Portugal
 European Commission Seventh Framework Programme for Research and Technological Development
Concluded
2012 - 2013 NANOFOL - Folate-based nanobiodevices for integrated diagnosis/therapy targeting chronic inflammatory diseases.
FP7-NMP-2008-LARGE-2
PhD Student Fellow
Universidade do Porto Instituto de Biologia Molecular e Celular, Portugal
 European Commission Seventh Framework Programme for Research and Technological Development
Concluded
2005 - 2006 Deciphering the mechanisms of rabbit genetic resistance to myxomatosis and rabbit haemorrhagic viral disease.
POCI/BIA-BDE/61553/2004
Integration into Research Grant Fellow
Universidade do Porto Instituto de Ciências Biomédicas Abel Salazar, Portugal
 Fundação para a Ciência e a Tecnologia
Concluded

Other

Designation Funders
2019 - Current R2Bone - Non-coding RNA-based therapies for bone regeneration.
POCI-01-0145-FEDER-031402
Researcher
Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
 Fundação para a Ciência e a Tecnologia
Ongoing
2006 - 2012 Development and validation of in vitro bioassays for thyroid hormone receptor mediated endocrine disruption.
ZonMW - NWO grant (11.400.0075) part of the Alternatives to Animal Experiments Program
Researcher
Wageningen Universiteit en Research, Netherlands
 ZonMw
Concluded
2004 - 2005 Controlling inflammatory responses in the airways.
Netherlands Asthma Foundation; contract grant number: 99.27.
Scientific Initiation Fellow
Amsterdam Universitair Medische Centra, Netherlands
 Long Fonds
Concluded
Outputs

Publications

Journal article
  1. Brás, João Paulo; Bravo, Joana; Freitas, Jaime; Barbosa, Mário Adolfo; Santos, Susana Gomes; Summavielle, Teresa; Almeida, Maria Inês. "TNF-alpha-induced microglia activation requires miR-342: impact on NF-kB signaling and neurotoxicity". Cell Death & Disease 11 6 (2020): http://dx.doi.org/10.1038/s41419-020-2626-6.
    10.1038/s41419-020-2626-6
  2. Moura, Sara Reis; Bras, Joao Paulo; Freitas, Jaime; Osório, Hugo; Barbosa, Mario Adolfo; Santos, Susana Gomes; Almeida, Maria Ines. "miR-99a in bone homeostasis: Regulating osteogenic lineage commitment and osteoclast differentiation". Bone 134 (2020): 115303. http://dx.doi.org/10.1016/j.bone.2020.115303.
    10.1016/j.bone.2020.115303
  3. Plácido, Alexandra; Bueno, João; Barbosa, Eder A.; Moreira, Daniel C.; Dias, Jhones do Nascimento; Cabral, Wanessa Felix; Albuquerque, Patrícia; et al. "The Antioxidant Peptide Salamandrin-I: First Bioactive Peptide Identified from Skin Secretion of Salamandra Genus (Salamandra salamandra)". Biomolecules 10 4 (2020): 512. http://dx.doi.org/10.3390/biom10040512.
    10.3390/biom10040512
  4. Freitas, Jaime; Santos, Susana Gomes; Gonçalves, Raquel Madeira; Teixeira, José Henrique; Barbosa, Mário Adolfo; Almeida, Maria Inês. "Genetically Engineered-MSC Therapies for Non-unions, Delayed Unions and Critical-size Bone Defects". International Journal of Molecular Sciences 20 14 (2019): 3430. http://dx.doi.org/10.3390/ijms20143430.
    10.3390/ijms20143430
  5. Oliveira, Marta S.; Freitas, Jaime; Pinto, Pedro A. B.; de Jesus, Ana; Tavares, Joana; Pinho, Mafalda; Domingues, Rita G.; et al. "Cell Cycle Kinase Polo Is Controlled by a Widespread 3' Untranslated Region Regulatory Sequence inDrosophila melanogaster". Molecular and Cellular Biology 39 15 (2019): http://dx.doi.org/10.1128/mcb.00581-18.
    10.1128/mcb.00581-18
  6. Carvalho, D.O.; Freitas, J.; Nogueira, P.; Henriques, S.N.; Carmo, A.M.; Castro, M.A.; Guido, L.F.. "Xanthohumol inhibits cell proliferation and induces apoptosis in human thyroid cells". Food and Chemical Toxicology 121 (2018): 450-457. http://www.scopus.com/inward/record.url?eid=2-s2.0-85054021358&partnerID=MN8TOARS.
    10.1016/j.fct.2018.09.021
  7. Braz, Sandra Oliveira; Cruz, Andrea; Lobo, Andrea; Bravo, Joana; Moreira-Ribeiro, Joana; Pereira-Castro, Isabel; Freitas, Jaime; et al. "Expression of Rac1 alternative 3' UTRs is a cell specific mechanism with a function in dendrite outgrowth in cortical neurons". Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms 1860 6 (2017): 685-694. http://dx.doi.org/10.1016/j.bbagrm.2017.03.002.
    10.1016/j.bbagrm.2017.03.002
  8. Nogueira, E.; Freitas, J.; Loureiro, A.; Nogueira, P.; Gomes, A.C.; Preto, A.; Carmo, A.M.; Moreira, A.; Cavaco-Paulo, A.. "Neutral PEGylated liposomal formulation for efficient folate-mediated delivery of MCL1 siRNA to activated macrophages". Colloids and Surfaces B: Biointerfaces 155 (2017): 459-465. http://www.scopus.com/inward/record.url?eid=2-s2.0-85019046361&partnerID=MN8TOARS.
    10.1016/j.colsurfb.2017.04.023
  9. Liu, X.; Freitas, J.; Zheng, D.; Oliveira, M.S.; Hoque, M.; Martins, T.; Henriques, T.; Tian, B.; Moreira, A.. "Transcription elongation rate has a tissue-specific impact on alternative cleavage and polyadenylation in Drosophila melanogaster". RNA 23 12 (2017): 1807-1816. http://www.scopus.com/inward/record.url?eid=2-s2.0-85034084061&partnerID=MN8TOARS.
    10.1261/rna.062661.117
  10. Nogueira, Eugénia; Cruz, Célia F.; Loureiro, Ana; Nogueira, Patrícia; Freitas, Jaime; Moreira, Alexandra; Carmo, Alexandre M.; et al. "Assessment of liposome disruption to quantify drug delivery in vitro". Biochimica et Biophysica Acta (BBA) - Biomembranes 1858 2 (2016): 163-167. http://dx.doi.org/10.1016/j.bbamem.2015.11.008.
    10.1016/j.bbamem.2015.11.008
  11. Nogueira, Eugénia; Mangialavori, Irene C.; Loureiro, Ana; Azoia, Nuno G.; Sárria, Marisa P.; Nogueira, Patrícia; Freitas, Jaime; et al. "Peptide Anchor for Folate-Targeted Liposomal Delivery". Biomacromolecules 16 9 (2015): 2904-2910. http://dx.doi.org/10.1021/acs.biomac.5b00823.
    10.1021/acs.biomac.5b00823
  12. Nogueira, E.; Lager, F.; Le Roux, D.; Nogueira, P.; Freitas, J.; Charvet, C.; Renault, G.; et al. "Enhancing Methotrexate Tolerance with Folate Tagged Liposomes in Arthritic Mice". Journal of biomedical nanotechnology 11 12 (2015): 2243-2252. http://www.scopus.com/inward/record.url?eid=2-s2.0-84941551653&partnerID=MN8TOARS.
    10.1166/jbn.2015.2170
  13. Freitas, J.; Miller, N.; Mengeling, B.J.; Xia, M.; Huang, R.; Houck, K.; Rietjens, I.M.C.M.; Furlow, J.D.; Murk, A.J.. "Identification of thyroid hormone receptor active compounds using a quantitative high-throughput screening platform". Current Chemical Genomics and Translational Medicine 8 1 (2014): 36-46. http://www.scopus.com/inward/record.url?eid=2-s2.0-84989270930&partnerID=MN8TOARS.
    10.2174/1389557515666150709104018
  14. Nogueira, Eugénia; Loureiro, Ana; Nogueira, Patrícia; Freitas, Jaime; Almeida, Catarina R.; Härmark, Johan; Hebert, Hans; et al. "Liposome and protein based stealth nanoparticles". Faraday Discussions 166 (2013): 417. http://dx.doi.org/10.1039/c3fd00057e.
    10.1039/c3fd00057e
  15. Gutleb, A.C.; Freitas, J.; Murk, A.J.; Verhaegen, S.; Ropstad, E.; Udelhoven, T.; Hoffmann, L.; Audinot, J.-N.. "NanoSIMS50 - A powerful tool to elucidate cellular localization of halogenated organic compounds". Analytical and Bioanalytical Chemistry 404 9 (2012): 2693-2698. http://www.scopus.com/inward/record.url?eid=2-s2.0-84870927176&partnerID=MN8TOARS.
    10.1007/s00216-012-6066-8
  16. Freitas, J.; Cano, P.; Craig-Veit, C.; Goodson, M.L.; David Furlow, J.; Murk, A.J.. "Detection of thyroid hormone receptor disruptors by a novel stable in vitro reporter gene assay". Toxicology in Vitro 25 1 (2011): 257-266. http://www.scopus.com/inward/record.url?eid=2-s2.0-78650522817&partnerID=MN8TOARS.
    10.1016/j.tiv.2010.08.013
  17. Muller, A.; Freitas, J.; Silva, E.; Le Gall-Reculé, G.; Zwingelstein, F.; Abrantes, J.; Esteves, P.J.; et al. "Evolution of rabbit haemorrhagic disease virus (RHDV) in the European rabbit (Oryctolagus cuniculus) from the Iberian Peninsula". Veterinary Microbiology 135 3-4 (2009): 368-373. http://www.scopus.com/inward/record.url?eid=2-s2.0-62149127401&partnerID=MN8TOARS.
    10.1016/j.vetmic.2008.09.057
  18. van den Berg, A.; Freitas, J.; Keles, F.; Snoek, M.; van Marle, J.; Jansen, H.M.; Lutter, R.. "Cytoskeletal architecture differentially controls post-transcriptional processing of IL-6 and IL-8 mRNA in airway epithelial-like cells". Experimental Cell Research 312 9 (2006): 1496-1506. http://www.scopus.com/inward/record.url?eid=2-s2.0-33646137375&partnerID=MN8TOARS.
    10.1016/j.yexcr.2006.01.010
Thesis / Dissertation
  1. Freitas, Jaime. "Development and validation of in vitro bioassays for thyroid hormone receptor mediated endocrine disruption. (ISBN 9789461734372)". PhD, Wageningen Universiteit en Research, 2012. https://library.wur.nl/WebQuery/wurpubs/431578.

Intellectual property

Patent
  1. Freitas, Jaime; Alexandra moreira; Ana Jesus; Marta Oliveira. 2019. "iPLUS, a nucleic acid comprising a 28-nucleotide sequence that activates gene expression and protein production in vitro and in vivo. (INIP 20181000064673)". Portugal.
    Pending
Activities

Oral presentation

Presentation title Event name
Host (Event location)
2019/02/01 Efficacy of GMP-graded FBN, in vivo testing – Blood biological parameters (FOLSMART M36 Steering Committee Meeting) FOLSMART M36 Steering Committee Meeting
MUW Medical University of Vienna (Viena, Austria)
2018/06/20 In vitro testing of GLP/GMP Grade materials, Genotoxicity studies (FOLSMART M30 Steering Committee Meeting) FOLSMART M30 Steering Committee Meeting
Aptuit (Verona, Italy)
2017/06/07 Effect of sulfasalazine (SSZ) in human activated macrophages (FOLSMART M18 Steering Committee Meeting) FOLSMART 3rd Steering Committee Meeting
MUW Medical University of Vienna (Viena, Austria)
2013 Folate-based nanodevice uptake specificity in FRß- expressing macrophages. (NANOFOL M42 Steering Committee Meeting) NANOFOL M42 Steering Committee Meeting
MUW Medical University of Vienna (Viena, Austria)
2012/06/01 Folate-based nanodevice uptake specificity in FRß- expressing macrophages. (NANOFOL M30 Steering Committee Meeting) NANOFOL M30 Steering Committee Meeting
INSERM (Paris, France)
2012 Folate-based nanodevice uptake specificity in FRß- expressing macrophages. (NANOFOL M36 Steering Committee Meeting) NANOFOL M36 Steering Committee Meeting
EXBIO (Praga, Czech Republic)
2009 Development and validation of a highly sensitive reporter gene assay for thyroid hormone receptor specific endocrine disruptors. (Conference) Dutch Society of Toxicology annual meeting
Dutch Society for Toxicology (Veldhoven, Netherlands)
2008 Detection of thyroid hormone receptor disruptors by a novel in vitro reporter gene assay. (Conference) SETAC Europe annual meeting
SETAC (Varsovia, Poland)

Supervision

Thesis Title
Role 		Degree Subject (Type)
Institution / Organization
2014 - 2019/07/23 Plk-1, a candidate regulator of RNAPII. (PhD thesis)
Co-supervisor
 Doctoral Program in Molecular and Cell Biology (PhD)
Universidade do Porto, Portugal
2016 - 2017 Upstream sequence element as a regulator of gene expression in different cellular contexts. (Licenciatura project)
Supervisor
 Licenciatura em Bioquímica (Other)
Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
2013 - 2014 Integrating RNA polymerase II dynamics with alternative polyadenylation. (MSc Thesis)
Co-supervisor
 MSc in Bioengineering (Master)
Universidade do Porto, Portugal
2013 - 2014 Unravelling the molecular mechanisms of polo alternative polyadenylation. (MSc thesis)
Co-supervisor
 MSc in Cell and Molecular Biology (Master)
Universidade do Porto, Portugal
2012 - 2013 Xanthohumol exerts chemopreventive effects on a papillary thyroid carcinoma cell line. (MSc thesis)
Co-supervisor
 MSc in Bioengineering
Universidade do Porto Instituto de Biologia Molecular e Celular, Portugal
2012 - 2012 Effect of fetal calf serum on cytotoxicity, size and cellular uptake of nanoparticles. (MSc thesis)
Co-supervisor
 MSC in Food Safety (Master)
Wageningen Universiteit en Research, Netherlands
2010 - 2010 Evaluation of compounds disrupting thyroid hormone receptor by using GH3.TRE-LUC cells. (MSc thesis)
Co-supervisor
 MSc in Toxicology (Master)
Wageningen Universiteit en Research, Netherlands
2010 - 2010 Effect of fetal calf serum on cytotoxicity, size and cellular uptake of nanoparticles. (MSc thesis)
Co-supervisor
 MSc in Toxicology (Master)
Wageningen Universiteit en Research, Netherlands
2009 - 2009 Development of a TRa.HELA-CMV-Luc cell line to enable the detection of Thyroid Receptor-isoform specific disruption. (MSc thesis)
Co-supervisor
 MSc in Toxicology (Master)
Wageningen Universiteit en Research, Netherlands
2008 - 2008 The effect of known thyroid hormone disrupting compounds on the newly developed GH3.TRE-Luc cell line (MSc thesis)
Co-supervisor
 MSc in Toxicology (Master)
Wageningen Universiteit en Research, Netherlands
2008 - 2008 New bioluminescence assay to detect thyroid hormone receptor disrupters. (MSc thesis)
Co-supervisor
 MSc in Toxicology (Master)
Wageningen Universiteit en Research, Netherlands
2007 - 2008 New detection system for thyroid hormone disrupters with a stably transfected GH3 cell line. (MSc thesis)
Co-supervisor
 MSc in Toxicology (Master)
Wageningen Universiteit en Research, Netherlands

Event organisation

Event name
Type of event (Role) 		Institution / Organization
2019/01/24 - 2019/01/25 VIII Portuguese Meeting: ptRNA2019. (Conference) (2019/01/24 - 2019/01/25)
Congress (Co-organisor)
 Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal

Jury of academic degree

Topic
Role 		Candidate name (Type of degree)
Institution / Organization
2018/10/17 Gene Regulation at the Chromatin Level in Arabidopsis: DNA-RNA hybrids formation at DOG1 promoter controls sense and antisense expression. (MSc thesis)
(Thesis) Main arguer
 Miguel Montez Mariano Coelho (Master)
Universidade do Porto, Portugal
2017/12/04 Cloning and molecular characterization of mouse S4DSRCRB. (MSc thesis)
(Thesis) Main arguer
 Helena Kalaitzopoulos (Master)
Universidade do Porto, Portugal
2014 Fst e Muc68E na resposta a chill coma e na resposta geral ao stress em Drosophila melanogaster. (Licenciatura project)
(Thesis) Main arguer
 João Rodrigues (Other)
Universidade do Porto, Portugal
2014 Frost e Muc68E como genes reguladores da resposta ao stress em Drosophila melanogaster. (Licenciatura project)
(Thesis) Main arguer
 Lisete Moutinho (Other)
Universidade do Porto, Portugal

Conference scientific committee

Conference name Conference host
2017/11/10 - 2017/11/10 6th i3S Annual Meeting Axis Vermar Conference & Beach Hotel in Póvoa de Varzim

Course / Discipline taught

Academic session Degree Subject (Type) Institution / Organization
2018/01/29 - 2018/02/02 Regulation of Gene Expression (GABBA PhD course) GABBA doctoral programme course (Curso de doutoramento (conclusão de unidades curriculares)) Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
2017/05/05 - 2017/05/09 Regulation of Gene Expression (GABBA PhD course) GABBA doctoral programme course (Curso de doutoramento (conclusão de unidades curriculares)) Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
2016/05/09 - 2016/05/13 Regulation of Gene Expression (GABBA PhD course) GABBA doctoral programme (Curso de doutoramento (conclusão de unidades curriculares)) Universidade do Porto Instituto de Investigacao e Inovacao em Saude, Portugal
2006 - 2012 Food Toxicology (MSc practical lectures) Food Toxicology (Mestrado) Wageningen Universiteit en Research, Netherlands
2006 - 2012 Environmental Toxicology (MSc practical lectures) Environmental Toxicology (Mestrado) Wageningen Universiteit en Research, Netherlands
2006 - 2012 Cell Biology & Health (MSc practical lectures) Cell Biology & Health (Mestrado) Wageningen Universiteit en Research, Netherlands