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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: 18d. Inventory of Institutions

Type of Institution: Hospital

Staff:

Damián García Olmo, Email: dgolmo@arrakis.es, Dr Medicina.
Dolores García Olmo, Email: logol@chospab.es, Ld Veterinaria.
Esperanza Martínez Navarro, Email: emarnav@chospab.es, Ld Biología.
Elena Navarro García, Email: eng_jcs@chospab.es, Ld Biología.
Jesús Ontañón Rodríguez, Email: jontanon@chospab.es, Ld Biología.

Total staff involved in alternative methods is 5 people.

Activities / aims: This hospital department is mainly involved in basic research; it performs cell biology, molecular biology and pathology testing on a routine basis, and surgery studies at a research level. Pharmaceuticals are occasionally studied.

Model systems: The model systems used are conventional animal models and in vitro methods -- cell lines culture (DHD/K12- rat colon tumor cell line).

Work lines: Colon adenocarcinoma-metastasis

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

Sources of financing: Fondo de Investigaciones Sanitarias (1995-97), Programa I+D en Salud (1997-98), Junta Comunidades Castilla-La Mancha (1995-97)

Type of Institution: Hospital

Staff:

Jorge Cannata Andia, Dr Medicina, Jefe de Servicio.
Carlos Gómez Alonso, Dr Medicina, Adjunto.
Jose Luis Fernández Martín, Ld Química, Adjunto, 50 % time devoted to altern. meth.
Jose Bernardino Díaz López, Ld Medicina, Adjunto.
Manuel Javes Díaz, Dr Biología, Adjunto, 75 % time devoted to altern. meth.
Carmen Diaz Corte, Ld Medicina, Becario, 25 % time devoted to altern. meth.
Ana Weruaga Rey, Lda Medicina, 25 % time devoted to altern. meth.
Primitiva Menendez, Dr Medicina, Adjunto, 25 % time devoted to altern. meth.

Total staff involved in alternative methods is 8 people.

Activities / aims: This hospital department is mainly involved in basic research; it performs cell biology, molecular biology, biochemistry, toxicology (nephrotoxicity) testing, pathology, quality control and culture methodology studies on a routine basis. Hormones, pharmaceuticals, diverse chemical compounds and drugs of abuse are routinely evaluated.

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

Model systems: The model systems used are conventional animal models (reduction, refinement) and in vitro methods -- culture of explants, reaggregates, reconstituted organs (femoral, neck, tibial), primary culture of dispersed cells (human osteoblasts) and cell lines culture (ROJ 17/28, M6-63).

The endpoints employed in vitro are cytoskeleton/membranes/enzyme release (alkaline phosphatase, osteocalcein), cellular proliferation (Tyd uptake), metabolic activity and nucleic acids (traduction).

Experimental systems (examples):

1 primary culture of osteoblasts, osteocalcein by RIA, effect of calcitriol, complementary test
2 M6-63 osteoblastic cell line culture, osteocalcein by RIA, effect of aluminum, replacement test
3 primary culture of osteoblasts, alkaline phosphatase, proliferation and osteocalceine, effect of aluminum, replacement test
4 rat parathyroid gland culture, PTH, RNApth, effect of aluminum, replacement test

Work lines: Renal osteodistrophy. Toxicology of aluminum. Bone metabolism.

Sources of financing: Hospital Central de Asturias, FIS, CICYT, Fundación Renal Iñigo Alvarez de Toledo

Type of Institution: Hospital

Staff:

Pablo Aldazabal, Email: paldaza@hnsa.es, Ld Medicina, Coordinador EUX


Total staff involved in alternative methods is 1 person.

Activities / aims: This hospital department is mainly involved in basic research and non-regulated applied research; it performs physiology, toxicology (acute systemic toxicity, ocular cytotoxicity, dermal irritation and corrosivity) and nutrition testing on a routine basis, and cell biology, biochemistry and pathology studies at a research level. Biomaterials, wastes, surgery are routinely evaluated; hormones and medical devices also being occasionally studied.

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

Model systems: The model systems used are conventional animal models, in vitro methods -- cell lines culture-- , human volunteers (clinical trials phases >III) and education models (animal, video, simulator, mechanical models).

The endpoints employed are in vivo (histopathological and physiological parameters) and in vitro morphology, cell signalling and defence systems.

Work lines: Experimental surgery, total parenteral nutrition in the rat, immunohystochemistry, neurogenetics, biomaterials.

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

Sources of financing: FIS, CICYT, Gobierno Vasco, Pharmaceutical Industry.

Type of Institution: Hospital

Staff:

Miguel Blanca Gómez, Email: mblanca@eurociber.es, Dr Medicina, Jefe Unidad, 20 % time devoted to altern. meth.
Cristobalina Mayorga Mayorga, Dr Biología, Becario Investigación , 20 % time devoted to altern. meth.
Manuel Rosal Sánchez, del, Dr Medicina, Becario Investigación , 30 % time devoted to altern. meth., duties related to altern. meth.: Inmunólogo
Sinfoniano Posadas Mañanes, Ld Químicas, Becario Investigación, 30 % time devoted to altern. meth., duties related to altern. meth.: Responsable Biología Molecular
Maria Jose Torres Jaen, Dr Medicina, Responsable Clínico (Adjunto), 40 % time devoted to altern. meth.

Total staff involved in alternative methods is 6 people.

Activities / aims: This hospital department is mainly involved in basic research, method development and method validation; it performs molecular biology, biokinetics, biotransformation and toxicology (mechanisms of toxicity, acute systemic toxicity, dermal irritation, corrosivity, immunotoxicity and sensitisation) testing on a routine basis, and monitoring --chemical, biological-- studies at a research level. Pharmaceuticals are routinely evaluated; cosmetics, diverse chemical compounds, biomaterials, food additives and colourings being also occasionally studied.

The main use of alternative methods is for replacement studies.

Model systems: The model systems used are in vitro methods - organ culture primary culture of cells (lymphocytes).

The endpoints employed in vitro are cellular proliferation, nucleic acids and organ-specific indicators.

Experimental systems (examples):

1 lymphocyte cell lines, biological response
2 lymphocyte cell lines, reaction

Work lines: In vitro methods for quantitating IgE antibodies to betalactams. Adverse reactions to drugs with immunological basis. Studies of the mechanisms involved. Methods for IgE production in vitro using peripheral cells from sensitized subjects.

Quality assurance / Validation programmes: The team have previously been involved in the following alternative method validation programmes: validation of RIA and ELISA methods. They are available to participate in EU validation programmes in relation to the evaluation of immunotoxicological methods, metabolic transformation, and detoxification mechanisms.

Sources of financing: Project Biomed, Project Biotech, Project FIS, CICYC, Consejeria Salud, Ministerio Educación y Ciencia, PAI

Others: General interest in the development of in vitro methods for the study of immunotoxicological reactions to pharmaceuticals and chemical compounds.

Type of Institution: Governmental research facility

Staff:

Maria Jose Gómez-Lechón, Email: mjgomez@san.gva.es, Dra Biología, Adjunto Investigación, 100 % time devoted to altern. meth., duties related to altern. meth.: Vocal Junta de ESTIV
Jose Vicente Castell Ripoll, Email: jose.castell@gong.ci.uv.es, Dr Química y Farmacia, Jefe de Sección, 100 % time devoted to altern. meth., duties related to altern. meth.: Consejo Científico ESTIV, Representante Español en ESAC-ECVAM
Ramiro Jover, Email: ramiro.jover@uv.es, Dr Biología, Contratado, 100 % time devoted to altern. meth.
Teresa Donato, Dr Farmacia, Adjunto Interino, 100 % time devoted to altern. meth.
Xavier Ponsoda, Dr. Biología, Colaborador.

Total staff involved in alternative methods is 11 people.

Activities / aims: This governmental research facility department is mainly involved in basic research, non-regulated applied research and method development; it performs toxicology testing on a routine basis, and cell biology, molecular biology, biochemistry, biokinetics and biotransformation and culture methodology studies at a research level. Pharmaceuticals, cosmetics, diverse chemical compounds, drugs of abuse, biomaterials and food additives are occasionally evaluated.

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

Model systems: The model systems used are in vitro methods --organ culture (liver), primary culture of dispersed cells (hepatocytes), cell lines culture (several hepatoma cell lines), cell-free systems (microsomes) and human volunteers (metabolism studies).

The endpoints employed in vitro are cell viability (MTT reduction, neutral red uptake, XTT reduction, LDH leakage, ATP), metabolic activity (liver), biotransformation systems (phase I [CYPA] and phase II), defence systems (GSH), organ-specific indicators (from liver [glucuronoconjugation, plasmatic proteins]) and toxicity mechanisms (Ca+2, lipid peroxidation).

Experimental systems (examples):

1 primary culture of hepatocytes from several species, hepatic physiological criteria, hepatotoxicity replacement test
2 primary culture of hepatocytes from several species, metabolic profile, HPLC/RMN, metabolism
3 transgenic cell lines expressing only one CYP, metabolism of specific substrate, identification of CYPS related to metabolism
4 primary culture of keratinocytes and fibroblasts, MTT reduction, phototoxicity, photodegradation

Work lines: Hepatotoxicity of drugs in cultured hepatocytes (several species). Mechanisms of hepatotoxicity. Evaluation of new drugs safety. Metabolism of drugs using primary cultured hepatocytes (including human) and CYP expressing transgenic cell lines. Development of stable and competent immortal hepatic cellular models.

Quality assurance / Validation programmes: The team have previously been involved in the following alternative method validation programme: MEIC. They are available to participate in EU validation programmes in relation to Biomed, Biotech, AIR, FLAIR.

Sources of financing: CEE (6 projects): AIR, Biomed I, Biomed II, Biotech; FIS (2); Pharmaceutical industry

Type of Institution: Hospital

Staff:

Fernando Yelamos Rodríguez, Dr Medicina, 35 % time devoted to altern. meth.
Francisco Laguez Britones, Dr Medicina, 25 % time devoted to altern. meth.
D.C. Martín Rubí, Ld Medicina, 15 % time devoted to altern. meth.
D.L. Blanco Coronado, Dr Medicina, 10 % time devoted to altern. meth.
Federico Orozco Rodríguez , 10 % time devoted to altern. meth.
Mª Carmen Marfil Montoya, Ld Medicina, 5 % time devoted to altern. meth.

Total staff involved in alternative methods is 6 people.

Activities / aims: This hospital department is mainly involved in regulatory testing; it performs toxicology (mechanisms of toxicity, basal cytotoxicity, acute systemic toxicity, ocular cytotoxicity, dermal irritation and corrosivity, nephrotoxicity, respiratory cytotoxicity, reproductive cytotoxicity, neurotoxicity, immunotoxicity and sensitisation, haematoxicity, carcinogenicity, phototoxicity, hepatotoxicity, chronic toxicity) and pathology studies on a routine basis. Pesticides are routinely evaluated.

Work lines: Acute and chronic effects of organophosphate pesticides

Type of Institution: Hospital

Staff:

Carmen Domínguez, Email: cdoming@hg.vhebron.es, Dr Biología, 30 % time devoted to altern. meth.
María Rosa Leiva, Diplomada en Enfermería, 80 % time devoted to altern. meth.
Eduard Hidalgo Albert, Ld Farmacia, 30 % time devoted to altern. meth.

Total staff involved in alternative methods is 3 people.

Activities / aims: This hospital department is mainly involved in non-regulated applied research and method development; it performs cell biology, biochemistry, toxicology (mechanisms of toxicity) and diagnostic (metabolopaties) studies on a routine basis and pharmacodynamics (cytotoxicity of antiseptics) studies at a research level. Pharmaceuticals and metals are occasionally evaluated.

Model systems: The model systems used are conventional animal models and in vitro methods -- culture of explants, reaggregates, reconstituted organs, (dermal fibroblasts) and cell lines culture (MDCK).

The endpoints employed in vitro are cell viability (MTT and XTT reduction, neutral red uptake) cytoskeleton /membranes/enzyme release (LDH, hexosaminidase), cellular proliferation (cell count, total protein content), metabolic activity (lysosomal, oxidative stress) and defence systems (antioxidative mechanisms).

Work lines: Study of cytotoxicity mechanisms of aluminum and lead in human dermal fibroblasts. Study of the most frequently applied topical antiseptics: antimicrobial activity related to cytotoxicity in cultured human fibroblasts. Growth modulating effects of radical oxygen species.

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes in relation to the techniques used

Sources of financing: Self financing, pharmaceutical industry.

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.