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Inventory of the Spanish Institutions and Scientists Involved in Alternatives to the use of Laboratory Animals (Refinement, Reduction or Replacement)+

Inventory: 18b. Inventory of Institutions

Type of Institution: Governmental research facility

Staff:

Margarita Alía Díaz, Email: arias@cc.csic.es, Dr Farmacia, Investigador Científico, 40 % time devoted to altern. meth.
José Antonio González Pérez, Dr Ciencias Biológicas, Investigador Contratado, 40 % time devoted to altern. meth.
Susana Cobacho Arcos, Ld Ciencias Biológicas, Becaria.
Lucia Irene Herencia Abendaño, Ingeniero Agrónomo, Becaria, 40 % time devoted to altern. meth.

Total staff involved in alternative methods is 4 people.

Activities / aims: This governmental research facility department is mainly involved in basic research, non-regulated applied research, method development and method validation; it performs toxicology testing (mechanisms of toxicity, reproductive cytotoxicity) on a routine basis and monitoring --chemical, biological-- studies at a research level. Pesticides and environmental pollutants are occasionally evaluated.

The main use of alternative methods is for complementary and replacement studies.

Model systems: The model systems used are animal models -- invertebrates (nematodes) and in vitro methods.
The endpoints employed are in vivo faunistic study of nematodes and in vitro cell viability (loss of sensitive groups).

Experimental systems (examples): 1 nematodes, order Dorylainmida, viability, soil contamination by nitrates, replacement test

Type of Institution: Governmental research facility

 Staff:

Eduardo de la Peña de Torres, Email: epena@ccma.csic.es, Dr C. Biológicas, 80 % time devoted to altern. meth.
Ana Guadaño Larrauri, Email: aguadano@ccma.csic.es, Dr C. Biológicas, 80 % time devoted to altern. meth.
Antonia Martínez López. Ayudante Diplomada CSIC.

Total staff involved in alternative methods is 3 people.

Activities / aims: This governmental research facility department is mainly involved in basic research; it performs toxicology testing (genotoxicity / mutagenicity) on a routine basis. Pesticides and new bioactive substances are routinely evaluated.

Model systems: The model systems used are in vitro methods -- micro-organisms (Salmonella typhimurium) and primary culture of dispersed cells (peripheral lymphocytes).

Sources of financing: CICYT- PGC, Ministerio de Agricultura

Type of Institution: Governmental research facility

Staff:

Jesús del Mazo Martínez, Email: cibjm26@fresno.csic.es, Dr Ciencias Biológicas, Jefe de Grupo, 90 % time devoted to altern. meth.
Luis Andrés López Fernández, Email: ciba257@fresno.csic.es, Dr Ciencias Biológicas, Becario Postdoctoral, 90 % time devoted to altern. meth.
Mario Párraga San Román, Email: m.parraga@fresno.csic.es, Dr Ciencias Biológicas, Becario Postdoctoral, 90 % time devoted to altern. meth.
Edmundo Bonilla González, Email: cibb106@fresno.csic.es, Ld Ciencias Biológicas, Becario Predoctoral, 90 % time devoted to altern. meth.
Fernando Escolar Antúnez, Email: cibea8s@fresno.csic.es, Técnico de Laboratorio, Personal Técnico, 90 % time devoted to altern. meth.

Total staff involved in alternative methods is 5 people.

Activities / aims: This governmental research facility department is mainly involved in basic research, non-regulated applied research, method development and method validation; it performs cell biology, molecular biology, toxicology (reproductive cytotoxicity), genetics, production (fertility) and culture methodology testing on a routine basis, and biochemistry studies at a research level. Pharmaceuticals, diverse chemical compounds, pesticides and environmental pollutants are routinely evaluated.

The main use of alternative methods is for screening and replacement test.

Model systems: The model systems used are conventional animal models and transgenics-, embryos (mouse) and in vitro methods --culture of explants, reaggregates, reconstituted organs (ovary), primary culture of dispersed cells (ovocytes, Sertoli cells, mouse sperm cells) and cell lines culture (15P-1 / Sertoli).

The endpoints employed are in vivo gene expression and in vitro nucleic acids and organ-specific indicators (stress proteins, gene expression).

Experimental systems (examples):

1 in vitro culture of germ cell lines, differential gene expression, gene expression markers

Work lines: Development of in vitro germ cell culture systems of and their use on reprotoxicity tests in mammals.

Quality assurance / Validation programmes: The team have previously been involved in the following alternative method validation programmes: Biotechnology

Sources of financing: UE, Comunidad Autónoma de Madrid, DGICyT

Type of Institution: Governmental research facility

Staff:

Cinta Porte Visa, Email: cpvqam@cid.csic.es, Dr Biología, Colaborador Cientifico, 50 % time devoted to altern. meth., duties related to altern. meth.: person in charge
Montserrat Sole Rovira, Email: msrqam@cid.cis.es, Dr Biología, Colaboradora Científica Contratada, 20 % time devoted to altern. meth.
Verónica Borghi, Email: vebqam@cid.csic.es, Ld Biología, Becaria Investigación, 80 % time devoted to altern. meth.
Amaya Albalai Rives, Email: aarqam@cid.csic.es, Ld Biología, Contratada, 80 % time devoted to altern. meth.

Total staff involved in alternative methods is 4 people.

Activities / aims: This governmental research facility department is mainly involved in basic research and method development; it performs toxicology testing (mechanisms of toxicity) and monitoring --chemical, biological-- on a routine basis. Pesticides and environmental pollutants are routinely evaluated.

The main use of alternative methods is for complementary studies.

Model systems: The model systems used are animal models -- invertebrates (mussel, fresh water crayfish),-- and in vitro methods - primary culture of dispersed cells (digestive glands, Mytilus and Prokambarus clarkii) and cell-free systems (microsomes of aquatic organisms).

The endpoints employed in vitro are cell viability (photometry), biotransformation systems (cyt P450 (EROD), GST), defence systems (antioxidative enzymes) and AChE inhibition.

Experimental systems (examples):

1 primary culture of mussel digestive glands and gills, eosin-y exclusion by photometry; Ache and carboxylesterases, effects of organophosphate pesticides, complementary test

2 primary culture of the fresh water crayfish Prokambarus clarkii, eosin-y exclusion by photometry; Ache and carboxylesterases, effects of organophosphate pesticides, complementary test

3 primary cultures of mussel digestive glands, antioxidant enzymes and GST by spectrophotometry, effects of the pesticide iragrol and possible detoxification, screening

4 primary culture of mussel, viability by spectrophotometry, toxicity of naphthalene sulphonates

Work lines: Study the bioaccumulation and biotransformation of organic contaminants in marine organisms. Identify potential biomarkers of exposure/effect in order to assess the impact of pollution in aquatic ecosystems. Development and use of primary cell cultures (bivalves and crustaceans) as a tool to investigate the toxicity and mechanism of action of pollutants in aquatic organisms.

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes.

Sources of financing: MOPT, Evaluación del impacto del vertido de petróleo del "Aegean Sea" en poblaciones de bivalvos de la costa de La Coruña mediante el uso de índices bioquímicos de estrés" (Marzo 93 - Abril 94); Comunidad Europea-Programa ENVIRONMENT "Biological markers of environmental contamination in marine ecosystems" EV5V-CT94-0550 (Julio 94 - Julio 96); Comunidad Europea -Programa MAST-II- "Risk assessment of organotin antifouling on key benthic organisms of European coastal habitats. MAS2-CT94-099" (Enero 95 - Enero 97); DGICYT "Indices bioquímicos de contaminación ambiental en ecosistemas marinos. Acción Integrada con Portugal.HP94-022" (Octubre 95 - Octubre 96); PLANICYT "Marcadores biológicos de contaminación ambiental en ecosistemas marinos. AMB95-1092-CE" (Octubre 95 - Octubre 96); PLANICYT "Bioacumulación y toxicidad de compuestos organoestánnicos en organismos bentónicos de la costa mediterránea. AMB95-1978-CE" (Enero 96 - Enero 97); FAO-MED POL-Pase II "Biomonitoring of pollution effects in the Barcelona and Valencia coastal areas" (Enero 96 - Enero 97); DGICYT "Cultivos celulares primarios como sistemas modelo para investigar el metabolismo y toxicidad de pesticidas organofosforados. Acción Integrada en el Reino Unido". HB1995-0056 (Abril 96 - Marzo 97); Comunidad Europea-Programa ENVIRONMENT "Biological Markers of environmental contamination in marine ecosystems. Biomar II ENV4-CT96-0300" (Julio 96 - Julio 98); CSIC/PAS "Utilización de proteínas de estrés como biomarcadores de contaminación en zonas costeras 1997/98"; PLANICYT "Marcadores Biológicos de contaminación ambiental en ecosistemas marinos. AMB 97-1800-CE" (Enero 98 - Enero 99).

Type of Institution: University

Staff:

Maria Cascales Angosto, Email: cascales@eucmax.sim.ucm.es, Dra. Farmacia, Investigador Científico CSIC
Carmen Díez Fernandez, Email: carmen.diez@jrc.it, Dra. Farmacia, Contrato C E ISPRA ECVAM, Italy
Asunción Zaragoza Castellano, Email: cascales@eucmax.sim.ucm.es, Ld. Farmacia, Becaria UCM
David Andres García, Email: cascales@eucmax.sim.ucm.es, Ld. Farmacia, Becario
Alberto Alvarez Barrientos, Dr. Biología, Técnico Citometría UCM

Total staff involved in alternative methods is 5 people.

Activities / aims: This university department is mainly involved in basic research; it performs cell biology, molecular biology, toxicology (hepatotoxicity) and culture methodology studies on a routine basis. Pharmaceuticals and drugs of abuse are routinely evaluated.

Model systems: The model systems used are animal models and in vitro methods -- primary culture of dispersed cells and cell-free systems.

The endpoints employed in vitro are cell viability, cellular proliferation, metabolic activity and biotransformation systems.

Work lines: Development of a liver specific in vitro model for the identification of non-genotoxic carcinogens and compounds with tumour promoting activity (Sandoz-Novartis). Oxygen free-radicals involved in the mechanisms of xenobiotic-induced hepatotoxicity in murins. Influence of age (FIS 95/0032/01). Modulation of hepatotoxicity by xenobiotic interactions. Ageing effect and influence of dietary antioxidants (CICYT PM 96-0010).

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes.

Sources of financing: SANDOZ (NOVARTIS), 1994; FIS 95/0032/01, 1995-1997; CICYT PM 96-0010, 1997-1999

Type of Institution: Governmental research facility

Staff:

Cristina Suñol Esquirol, Email: csenqi@cid.csic.es, Dr Ciencias, Ld Ciencias Químicas, Ingeniero Químico IQS, Titulado , 80 % time devoted to altern. meth.
Elena Fonfria Subiró, Email: efsnqi@cid.csic.es, Ld Farmacia, Becaria, 100 % time devoted to altern. meth.
Total staff involved in alternative methods is 2 people.

Activities / aims: This governmental research facility department is mainly involved in basic research; it performs toxicology (mechanisms of toxicity, neurotoxicity) testing on a routine basis. Pesticides and environmental pollutants are routinely evaluated.

Model systems: The model systems used are in vitro methods - primary culture of dispersed cells (neuronal cells, rat/mouse).

The endpoints employed are in vitro cell viability (LDH, MTT reduction) and cell signalling (receptor binding, function, neurotransmitter release).

Experimental systems (examples):

1 primary culture of neurones, uptake and release of neurotransmitter (HPLC, and isotopes); receptor binding and ionic flux, neurotoxicity assays

Work lines: The use of in vitro neural systems to study mechanisms of neurotoxicity of environmental xenobiotics in relation to their interaction with specific neural activities, such as uptake and release of neurotransmitters and action through receptors (especially receptors for inhibitory neurotransmitters, like GABA and glycine) by determining their status and function (interaction with binding sites and effects on channels or messenger pathways).

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes in relation to pharmaceutical and toxic agents evaluation in neural in vitro systems

Sources of financing: CE-BIOMED, FIS, CICYT, pharmaceutical industry.

Type of Institution: Governmental research facility

Staff:

Coral Sanfeliu Pujol, Email: cspfat@cid.csic.es, Dr Biología, Colaborador Científico, 90 % time devoted to altern. meth.
Rosa Cristofol Martínez, Email: rcmfat@cid.csic.es, Dr Biología, Colaborador Científico, 100 % time devoted to altern. meth.
Eduardo Rodríguez-Farre, Dr Medicina, Investigador Científico, 25 % time devoted to altern. meth.
Sergi Gasso Pons, Email: sgpfat@cid.csic.es, Ld Biología, Becario Predoctoral, 100 % time devoted to altern. meth.
Jordi Sebastia Palleja, Email: jspfat@cid.csic.es, Ld Biología, 100 % time devoted to altern. meth.

Total staff involved in alternative methods is 7 people.

Activities / aims: This governmental research facility department is mainly involved in basic research and method development; it performs toxicology (mechanisms of toxicity, neurotoxicity) and culture methodology testing on a routine basis, and cell biology and pharmacology studies at a research level. Pesticides, environmental pollutants, neurotransmitters and neuroactive pharmaceuticals are routinely evaluated; pharmaceuticals also being occasionally studied.

The main use of alternative methods is for replacement studies.

Model systems: The model systems used are in vitro methods -- organ culture (rat brain slices), primary culture of dispersed cells (neurones, astrocytes, microglia (rat, mice, human)) and cell lines culture (neuroblastoma, PC12).

The endpoints employed in vitro are cell viability (propidium iodide, calcein), metabolic activity (MTT reduction, enzyme activities), nucleic acids (apoptosis detection) and organ-specific indicators (neurotransmitter release, oxidative stress).

Experimental systems (examples):

1 primary culture of neurones and/or astrocytes, membrane permeability to propidium iodide, cell viability and pharmaceutical protection, replacement test
2 primary culture of neurones and/or astrocytes, MTT reduction, cell viability and pharmaceutical protection, replacement test
3 primary culture of neurones, oxygen free-radical production, by oxidation of DCFH-DA, oxidative stress, replacement test
4 brain slices, neurotransmitter release, synaptic function, replacement test

Work lines: Neurotoxicity mechanisms of xenobiotics in neuronal cultures

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes in relation to neurotoxicity tests

Sources of financing: "Evaluación de la Neurotoxicidad selectiva y de los mecanismos de acción de xenobioticos ambientales en modelos neurales in vitro" FIS, Ministerio Sanidad y Consumo (No, 97/0656) 1997-1999; "Desarrollo de sistemas neurales in vitro para el estudio de los efectos tóxicos de xenobioticos"- FIS, Ministerio Sanidad y Consumo (No, 95/1955) 1995/1996; "Factores de Neurotoxicidad selectiva de xenobioticos" Comisión Interministerial de Ciencia y Tecnología (No SAF 94-0076) 1994-1997; "Development of in vitro neural and related immune systems for the study of potentially toxic effects of novel and highly specific compounds during cell differentiation" Programa BIOTECHNOLOGY Commission de Les Comunitats Europees (No BIOT-CT93-0224) 1994-1996. "Ajut per a potenciar els grups de qualitat en el marc del pla de recerca de Catalunya. Comissio Interdepartamental de Recerca i Innovacio Tecnologica (1995GR-00551) 1995; "Desarrollo de sistemas neurales in vitro para la identificación de agentes con potencial neurotoxicológico y farmacológico". FIS Ministerio de Sanidad y Consumo (Nº 93/0899E) 1993-1994. "Caracterización de mecanismos de acción y efectos neurotóxicos de policlorocicloalcanos" Comisión Interministerial de Ciencia y Tecnología (No SAL91-0707) 1992-1994.

Type of Institution: Governmental research facility

Staff:

Jaime Sancho Lopez, Email: granada@ipb.csic.es, Dr. Ciencias Biológicas, Jefe del Laboratorio y del Departamento.
Mercedes Zubiaur Marcos, Email: mzubiaur@ipb.csic.es, Dra. Ciencias Biológicas, Investigador Contratado.
Maria Guirado, Email: mguirado@ipb.csic.es, Ld. Farmacia, Becaria Predoctoral
Teresa Orta, Email: tereorta@ipb.cisc.es, Ld. Medicina, Becaria Predoctoral

Total staff involved in alternative methods is 4 people.

Activities / aims: This governmental research facility department is mainly involved in basic research; it performs cell biology, molecular biology and biochemistry testing on a routine basis, and pharmacodynamics (inhibition of cell physiological responses) and pathology studies at a research level.

The main use of alternative methods is for replacement studies.

Model systems: The model systems used are conventional animal models (immunization), transgenics (experimental model of diseases) and in vitro methods -- micro-organisms (bacteria) and cell lines culture (T lymphocyte, fibroblast, myeloid cell lines).

The endpoints employed in vitro are cell viability (trypan blue exclusion), cellular proliferation (cell count), cell signalling (tyrosine phosphorylation, intracellular associations, cytoquines), nucleic acids (transfection of kinases and tyrosine-kinases) and flow cytometry (FACS).

Experimental systems (examples):

1 human Jurkat T lymphocytes, tyrosine phosphorylation by immunoprecipitation, western blot, identification of new routes of activation and components

2 cos monkey fibroblasts, cDNAs transfection by FACS, western blot, immunofluorescence, possible interactions and biological effects

3 human Jurkat T lymphocytes cytokines, transcription factors by CAT, ELISA, gel-shift, immunoregulation studies

Work lines: Strategies for blocking the association of signalling proteins containing tandem SH2 domains with an activation motif present in T cells: in some systems, phosphatidilinositol 3-kinase (PI 3-K) constitutes the major link in the control of DNA synthesis. PI 3-K comprised a p85 regulatory subunit coupled to a p110 catalytic subunit. p85 contains two SH2 domains in tandem and constitutes a putative candidate for interacting with doubly phosphorylated ITAMs. CD38-mediated activation of tyrosine kinase signalling pathways: the lab has characterized some of the early signalling events triggered by CD38 engagement in Jurkat T cells. CD38 ligation with the specific mAb IB4 induced rapid and transient tyrosine phosphorylation of different patterns of cytoplasmic proteins, including PLC-1, c-Cbl, ZAP-70, Shc, and Erk-2 Mitogen-activated Protein (MAP) kinase.

Quality assurance / Validation programmes: The institution applies Good Laboratory Practices rules according to the the EU. The team are available to participate in EU validation programmes, in relation to Biomed and Biotech.

Sources of financing: "Identificación y clonaje de proteinas intracelulares asociadas al complejo TCR/CD (PB92-0123)", DGICYT; "Estructura y función de los receptores para la fracción Fc de la IgA (94/0666)", FIS; "Estudio de los antígenos HLA en tumores humanos y del complejo CD3-TCR en TIL y linfocitos autólogos (Expte: 95/1505)", FIS; "Estrategia para bloquear la asociación de proteínas que contienen dominios SH2 en tandem con un motivo de activación presente en los linfocitos T (SAF96-0117)", CICYT; "Requerimientos moleculares para la inducción de proliferación y activación celular mediados por CD38. Posible asociación con otros receptores y consecuencias funcionales (Expte: 96/49)", Consejeria de Salud. Junta de Andalucía; Consejería de Salud. Junta de Andalucia. Ayuda de apoyo a grupos de investigación. "Interacción de CD38 con el receptor de alta afinidad para la IgG, CSIC y CNR. "Reconstitution of interactions between signaling molecules and the TCR/CD3 complex, NATO; "Requerimientos moleculares de la señalización mediada por CD38" CSIC, CNR; "Señalización mediada por CD38 e identificación de las kinasas asociadas, CSIC e INSERM; "Señalización del receptor para el antígeno", CSIC y CNRS.

Type of Institution: University

 Staff:

Paulino Garcia Partida, Dr, Catedrático de Universidad.

Total staff involved in alternative methods is 1 person.

Activities / aims: This university department is mainly involved in basic research and alternatives to animals in education; it performs toxicology and pathology testing on a routine basis, and toxicology (nephrotoxicity, carcinogenicity, hepatotoxicity) and surgery studies at a research level. Pharmaceuticals are routinely evaluated; biomaterials and environmental pollutants also being occasionally studied.

The main use of alternative methods is for screening.

Model systems: The model systems used are conventional animal models and transgenics, and education models

Work lines: Education and training of scientists using experimental animals. Experimental pathology (oncology, toxicology, metabolism). Primates, alpha-farm-animals.

Quality assurance / Validation programmes: The team have previously been involved in alternative method validation programmes

Fuente / Source: Guillermo Repetto, Ana del Peso, Manuel Salguero, Manuel Repetto (1999) Inventory of the Spanish Institutions and Scientists Involved in Alternatives to the use of Laboratory Animals (Refinement, Reduction or Replacement) Revista de Toxicología 16: 50-127.