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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: 18k. Inventory of Institutions

Type of Institution: University

Staff:

C. Oscar Pintado Sanjuán, Email: cpiea@svq.servicom.es, Dr Veterinaria, Director Técnico del Servicio de Animales de experimentación, duties related to altern. meth.: Member of ethical Commitee Universidad de Sevilla

Total staff involved in alternative methods is 1 person.

Activities / aims: This University department is mainly involved in basic research; it performs on a routine basis cell biology, molecular biology, biochemistry, pharmacodynamics, pharmacology, physiology, biokinetics and biotransformation, toxicology and surgery studies and at a research level toxicology (genotoxicity / mutagenicity, carcinogenicity, hepatotoxicity, endocrine disruption, chronic toxicity), genetics, production, nutrition, diagnostics and monitoring -chemical, biological- studies.

Model systems: The model systems used are conventional animal models (rat, mouse and rabbit SPF), transgenics- and in vitro methods - cell lines culture (embryo).

Work lines: Animal breeding, laboratory animal pathology, policlonal antibodies, monoclonal antibodies, transgenic, knock out, wistar rat, WKY rat, SHR rat, C57BL/6 mouse, Swiss mouse, rabbit.

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

Sources of financing: New centre created with Fondos FEDER, University, and for research projects.

Others: Interest in alternative methods applied to basic research, for the promotion of alternatives in the animal room facilities of the university.

Type of institution: University

Staff:

Joaquín Piñero Bustamante, Email: pinero@cica.es, Dr Biología, Profesor Titular de Universidad, Biología Celular, 50 % time devoted to altern. meth.
Trinidad Ortiz Salles, Email: salles@cica.es, Dr Biología, Profesor Titular de Biología Celular, 50 % time devoted to alt. met.
Felipe Cortés Benavides, Email: cortes@cica.es, Dr Biología, Catedrático de Biología Celular, 50 % time devoted to altern. meth.
Santiago Mateos Codero, Email: smateo@cica.es, Dr Biología, Profesor Asociado Univers., 50 % time devoted to altern. meth.
Inmaculada Dominguez García, Email: idomin@cica.es, Dr Biología, Profesor Asociado de Univ., 50 % time devoted to altern. meth.

Total staff involved in alternative methods is about 5 persons.

Activities / aims: This university department is mainly involved in basic research; it performs cell biology and toxicology (genotoxicity / mutagenicity, carcinogenicity) studies on a routine basis. Diverse chemical compounds and physical agents are routinely evaluated, being environmental pollutants also occasionally studied.

The main use of alternatives methods is for complementary studies.

Model systems: The model systems used are in vitro methods -- cell lines culture (radiosensitives, tumor, wild and mutants).

The endpoints employed in vitro are cell viability (SRB, flow cytometry), cellular proliferation (cell count, flow cytometry), nucleic acids (degradation, apoptosis) and comet assay.

Experimental systems (examples):

1 radiosensitive and wild cell lines, RX, comet assay, lesion and repair, complementary test
2 cell lines, lesions by restriction enzymes, pulsed field electrophoresis, repair mechanisms, complementary test
3 cell lines, chromosomal aberrations, DNA lesions and repair, complementary test 

Type of Institution: University

Staff:

Ana Mª Cameán Fernández, Email: camean@fafar.us.es, Dr Farmacia, Profesor Titular Universidad, 30 % time devoted to altern. meth.
Olga Caneo López, Email: caneo@fafar.us.es, Ld Química, Becaria , 30 % time devoted to altern. meth.
Manuel Repetto Jiménez, Dr Ciencias y Medicina, Profesor Universidad, 10 % time devoted to altern. meth.

Total staff involved in alternative methods is 3 people.

Activities / aims: This university department is mainly involved in basic research; it performs toxicology (hepatotoxicity) testing on a routine basis. Toxins and environmental pollutants are occasionally evaluated.

The main use of alternative methods is for complementary studies.

Model systems: The endpoints employed in vitro are cell viability (MTT), and cytoskeleton/membranes/enzyme release (LDH release).

Work lines: Development of analytical methods for the determination of trace metals in biological and environmental samples, food and beverages. Development of chromatographic methods for the determination of toxins from cyanobacteria algae in water and other matrices (fish, molluscs). In vitro assays to study the toxic properties of microcystins.

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The team have previously been involved in alternative method validation programmes. They are available to participate in EU validation programmes.

Sources of financing: Toxicología de metales, Junta de Andalucia; Aplicaciones de las estratégicas quimioterápicas y en especial de las funciones hiperbólicas al estudio de los equilibrios simultáneos y estimación y optimización de parámetros de relevancia y significación analítica y biomédica, CICYT; Efecto de la Hipertensión arterial sobre los sistemas de transporte de monosacáridos en intestino y riñón, FIS; Desarrollo y aplicación de métodos químicos para la determinación de microcistinas en aguas destinadas al consumo, EMASESA; Fundación Avenzoar.

Type of Institution: University

Staff:

Francisco Javier Bedoya Bergua, Email: bedoya@cica.es, Dr Medicina, Prof. T.U, 100 % time devoted to altern. meth.
Remedios Ramírez Cárdenas, Dra Farmacia, Prof. Escuela Universitaria , 100 % time devoted to altern. meth.
Julio César Bernabé Ortiz, Ld Biología, Doctorando, 100 % time devoted to altern. meth.
Juan Tejedo Huamán, Ld Biología, Doctorando , 100 % time devoted to altern. meth.
Gladys Cahuana Maudo, Ld Biología.

Total staff involved in alternative methods is 5 people.

Activities / aims: This university department is mainly involved in non-regulated applied research; it performs molecular biology and biochemistry testing on a routine basis.

Model systems: The model systems used are in vitro methods -- cell lines culture (RINm5F cells).

The endpoints employed are in vitro cell viability (cell death).

Experimental systems (examples):

1 RINm5F cell line insuline producers, protein expresssion studied by immunoblotting, cell death, complementary test

Work lines: Mechanisms involved in the destruction of pancreatic ß-cells by the immune system

Sources of financing: FIS, DGCYT

Type of Institution: University

Staff:

Virginia Motilva Sánchez, Email: motilva@fafar.us.es, Dr Farmacia, Ld Biología, Profesor Titular

Total staff involved in alternative methods is 1 person.

Activities / aims: This university department is mainly involved in basic research; it performs molecular biology and pharmacological studies at a research level. Pharmaceuticals are routinely evaluated.

Model systems: The model systems used are conventional animal models and in vitro methods -- cell lines culture (CACO II colon cells).

Work lines: Application of natural products in gastric digestion pathology. Mechanisms of pharmaceutical gastric protection.

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

Sources of financing: "Investigación y desarrollo de nuevos fármacos útiles en patología digestiva" Programa ALFA Comisión Europea, Dirección General IB., Preproyecto; Subprograma B2 Relaciones exteriores: América Latina nº 4,0108,7, Nombre de la Red: RIMEPAD. 1997-1998; "Investigación y desarrollo de nuevos fármacos útiles en patología digestiva" Programa ALFA Comisión Europea, Dirección General IB., Preproyecto; Subprograma B2 Relaciones exteriores: América Latina nº6,0210,8, RIMEPAD 1998-1999; "Mechanisms involved in the gastroprotective effect mediated by Ibuprofen/L-arginine" Proyecto de Investigación Universidad de Sevilla-Laboratorios Zambon S.A. (Zambon Group), 1996-1998; "Mecanismos patogénicos de las lesiones gastrointestinales inducidas por dexketoprofeno vs ketoprofeno racémico. Modificaciones de las funciones absortivas del intestino delgado proximal" 1997-1998; "Mecanismos implicados en los efectos de metamizol sobre la mucosa gástrica" Proyecto de investigación Universidad de Sevilla-Laboratorios Boehringer Ingelheim, 1997-1998; Programa de Investigación y desarrollo Tecnológico de la Junta de Andalucía. Grupo de Investigación, Facultad de Farmacia Universidad de Sevilla. Area 30, Código 3119, 1988-92 y 1993-94; Programa de Investigación y Desarrollo Tecnológico de la Junta de Andalucía. Grupo de Investigación Mecanismos implicados en la gastropatia yatrogénica. Facultad de Farmacia Universidad de Sevilla. Area 30 Código CTS259, 1995-1997.

Type of Institution: University

Staff:

Maria José Medina Hernández, Email: maria.j.medina@uf.es, Dr, Profesor Titular Universidad, 80 % time devoted to altern. meth., duties related to altern. meth.: Investigador Principal Proyecto CICyT
Rosa María Villanueva Camañas, Email: rosa.m.villanueva@uf.es, Dr, Profesor Titular Universidad, 80 % time devoted to altern. meth., duties related to altern. meth.: Investigador Proyecto Investigación CICyT
Salvador Sagrado Vives, Email: salvador.sagrado@uf.es, Dr, Profesor Titular Universidad, 80 % time devoted to altern. meth., duties related to altern. meth.: Investigador Proyecto Investigación CICyT
Yolanda Martín Biosca, Email: yolanda.martin@uf.es, Dr, 80 % time devoted to altern. meth., duties related to altern. meth.: Investigador Proyecto Investigación CICyT
Jose María Sanchis Mallol, Dr, 50 % time devoted to altern. meth.
Laura Escuder Gilabert, Doctoranda, 100 % time devoted to altern. meth.
Mónica Molero Monfort, Doctoranda, 100 % time devoted to altern. meth.
Carmen Quiñones Torrelo, Doctoranda, 100 % time devoted to altern. meth.

Total staff involved in alternative methods is 8 people.

Activities / aims: This university department is mainly involved in basic research, non-regulated applied research and method development; it performs biokinetics and biotransformation and toxicology testing on a routine basis. Pharmaceuticals are routinely evaluated.

Model systems: The model systems used are mathematical modelling (liner/non-linear regression, chromatographic retention).

The endpoints employed in vitro are studied by HPLC

Experimental systems (examples):

1 micellar liquid , chromatography

Work lines: Quantitative retention-structure (hydrophobicity), retention-activity, retentition-pharmacokinetic studies by micellar liquid chromatography. Development and validation of empirical models used in prediction of new molecule properties/activities.

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

Sources of financing: Proyecto CICyT Plan Nacional de Salud y Farmacia: SAF 96-1709

Type of institution: University

Staff:

José Enrique O´Connor Blasco Email: jose.e.oconnor@uv.es, Dr Farmacia, Profesor Titular de Universidad, 50 % time devoted to altern. meth.
Guadalupe Herrera Martín, Email: jose.e.oconnor@uv.es, Lda en Ciencias Biológicas, Becaria de Investigación, 100 % time devoted to altern. meth.
Efrain Huerga Pérez, Ldo en Bioquímica, Becario de Investigación, 50 % time devoted to altern. meth.

Total staff involved in alternative methods is about 3 persons.

Activities / aims: This university department is mainly involved in basic research, non-regulated applied research and development of methods; it performs cell biology, biochemistry, toxicology (mechanisms of toxicity, basal cytotoxicity, neurotoxicity, genotoxicity / mutagenicity) testing on a routine basis, and diagnostics (immunological or coagulation alterations) and monitoring --chemical, biological- studies at a research level. Diverse chemical compounds are routinely evaluated; pesticides and toxins are also ocassionally evaluated.

The main use of alternatives methods is for screening, complementary and replacement studies.

Model systems: The model systems used are in vitro methods -- micro-organisms (WP2 modified E coli), primary culture of dispersed cells (human and rat isolated lymphocytes), cell lines culture (N13 rat hepatoma, Neuro-2a mouse neuroblastoma cells, L-132 human lung fibroblasts, human Jurkat leukemic, MOLT-4) and cell-free systems (isolated nuclei or mitochondria).

The endpoints employed are in vitro morphology (flow cytometry, laser dispersion), cell viability (flow cytometry by fluorochromes uptake), cytoskeleton / membranes / enzyme release (flow cytometry), celular proliferation (flow cytometry by DNA quantification and BrDU uptake), metabolic activity (mitochondrial activity, lipids, free-radicals), cell signalling (Ca+2 movements, pH, membrane potential), nucleic acids (aneuploidia, micronuclei, mutagenesis, SCE) and biotransformation systems (gluthation, GST, CYP450).

Experimental systems (examples):

1 WP2 E coli with mutations on oxidative stress control genes, H2O2 intracellular levels, gluthation, detection and quantification oxidative stress, screening and replacement test
2 cell lines culture: Neuro-2a, N13 rat hepatoma, cell viability, cytoplasmic and mitochondrial membrane potential, intracellular calcium levels, lipids, superoxide, peroxide, GSH, cell cycle, DNA ploidia, in vitro cytotoxicity (neurotoxicity, hepatotoxicity), screening and replacement studies
3 human lymphocytes culture, early (CD69) or late (CD25) antigen expression, adhesion molecules, flow cytometry, in vitro or ex vivo immunotoxicity study, screening and complementary studies

Work lines: The group is involved in the development of flow cytometric assays based upon functional parameters (signaling, metabolism, cell proliferation) to detect and quantify cytotoxicity using prokaryotic and eukariotic organisms as indicators / targets.

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

Sources of financing: Fondo de Investigaciones Sanitarias, Consellería de Educación, Izasa, S. A, C. E. Durviz, S.L, Ingelheim Diagnóstica y Tecnológica 

Type of Institution: University

Staff:

M. Anunciación Lafuente Giménez, Email: lafuente@uvigo.es, Dr CC Biológicas, Profesora Titular de Toxicología.
Nuria Márquez Almuiña, Email: nmarquez@uvigo.es, Ld Ciencia y Tecnología de los Alimentos, Becaria, Doctoranda.
Susana Piquero Villar, Email: spiquer@uvigo.es, Ld Ciencia y Tecnología de los Alimentos, Doctoranda.
Rubén Seara Escudero, Ld CC Químicas, Doctorando.

Total staff involved in alternative methods is 4 people.

Activities / aims: This university department is mainly involved in basic research; it performs toxicology (immunotoxicity and sensitisation, endocrine disruption) testing on a routine basis and biokinetic and biotransformation studies at a research level. Hormones and metals are routinely evaluated.

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

Model systems: The model systems used are conventional animal models and in vitro methods -- primary culture of dispersed cells (hepatocytes, splenocytes).

The endpoints employed are in vivo parameters for characterizing the secretion of hormones from adenohypophysis, catecholamines and indolamine levels and their metabolites in different brain regions and in vitro cell viability (LDH, trypan blue), cellular proliferation (ODC) and organ-specific indicators (lymphocyte phenotype).

Experimental systems (examples):

1 rat lymphocyte culture, lymphocyte subpopulations by flow cytometry, cytokines and interferon, by ELISA, immunotoxicity of heavy metals and pesticides, replacement test
2 primary cultures of trout hepatocytes, cell viability by trypan blue, and LDH leakage, protection of Zn and Se of the toxicity produced by Cd, complementary test
3 primary cultures of trout hepatocytes, determination of metabolites by HPLC with BR detector., study of biotransformation of xenobiotics, complementary test

Work lines: Neuroendocrine toxicology, immunotoxicology, toxicology of metals.

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes in relation to the V Programme on Health and Nutrition

Sources of financing: "Mejora de la producción piscícola: influencia de la contaminación medioambiental por cadmio" Fundación La Caixa, 1995; "Estudio in vitro de la acumulación de xenobioticos medioambientales utilizando el hepatocito como modelo cellular", Conselleria de Educación y Ordenación Universitaria de la Xunta de Galicia, 1992-1994 "Cronofarmacología de la ciclosporina", Sandoz, Pharma, SAE, 1994-1995 ; Liberación de octreotida en el sistema nervioso central" Sandoz, Pharma, SAE 1994-1995; Mecanismo de acción del interferon gamma en los ganglios linfáticos submaxilares. Papel de la inervación autonómica" DGICYT PB-94-260, 1995-1998; Efectos del cadmio sobre la secreción adenohipofisaria: papel del sistema inmunológico y neuromoduladores hipotalámicos" Xunta de Galicia, 1996-1998

Type of Institution: University

Staff:

Berta Rodríguez Campíns, Email: spinola@cece.es, Ld Ciencias Biológicas, Prof. Titular y Responsable del área de Toxicogía.
Iñigo Fajardo López-Cuervo, Email: spinola@cece.es, Dr Zoología y Biólogo, Jefe del Departamento de Biodiversidad Animal.
Virginia Pividal García, Email: spinola@cece.es, Ld Ciencias Biológicas, Especialidad Zoología, Jefe del departamento de Medio Natural.
Justo Martín Martín, Email: spinola@cece.es, Biólogo Especialidad Botánica y Zoología, Profesor Titular.
José Rafael Garrido López, Email: spinola@cece.es, Ld. Biología. Especialidad Zoología, Profesor Titular.
Manuela Segovia, Email: spinola@cece.es, Ld. Estadística, Responsable del área de análisis estadísticos.
Manuel Jiménez López, Ldo Veterinaria, Responsable del área de patología.

Total staff involved in alternative methods is 7 people.

Activities / aims: This university department is mainly involved in regulatory testing and alternatives to animals in education; it performs toxicology (neurotoxicity, ecotoxicity), pathology, diagnostics, monitoring --chemical, biological studies at a research level. Pesticides, toxins and environmental pollutants are routinely evaluated; diverse chemical compounds and wastes also being occasionally studied.

Model systems: The model systems used are invertebrate conventional animal models, vegetables, in vitro methods -- micro-organisms, education models and mathematical modelling.

The endpoints employed in vivo are applied to sea water invertebrates and birds of prey.

Work lines: The team is basically focussed on the long term effects of pesticides and pollutants in the ecosystem, and would like to restrict the research to protected areas or utterly degraded zones.

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

Sources of financing: Self financing

Others: The team has just started work, after preparing the study basis.

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.