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Reducing suffering in experimental autoimmune encephalomyelitis (EAE)

This report is based on discussions and submissions from an expert working group consisting of veterinarians, animal care staff and scientists with expert knowledge relevant to the field. It aims to facilitate the implementation of the Three Rs (replacement, reduction and refinement) in the use of animal models or procedures involving experimental autoimmune encephalomyelitis (EAE), an experimental model used in multiple sclerosis research. The emphasis is on refinement since this has the greatest potential for immediate implementation. Specific welfare issues are identified and discussed, and practical measures are proposed to reduce animal use and suffering. Some general issues for refinement are summarised to help achieve this, with more detail provided on a range of specific measures to reduce suffering. © 2013 Elsevier Inc.

Cuesta Arriba. Antecedentes epistemológicos y metodológicos para una Evaluación Ambiental Estratégica -EAE- en la ejecución de Políticas Planes y Programas ambientales en el Area del Gran Catamarca -AGC-

En esta comunicación se informa sobre estudios socio ambientales realizados desde las perspectivas epistemológica y metodológica, en el Area del Gran Catamarca. El propósito es contribuir a una Evaluación Estratégica Ambiental. Se examinan antecedentes y se identifica la percepción de la problemática ambiental en diferentes sectores de población.. Para la construcción del Marco teórico se analizaron información escrita (libros, revistas periódicos) y webgrafia e infografia, de origen local, nacional e internacional. El abordaje transversal de la investigación revela la complejidad epistemológica del objeto de estudio cuando se busca alcanzar una visión contemporánea, global y contextualizadora del problema. Complejidad que se evidencia en los resultados de los análisis, de contenidos, multicriterial y hermeneutico. Las indagaciones sobre percepción de la problemática ambiental recogieron testimonios orales, cuestionarios y encuestas cuyas características se describen en el trabajo. Integrantes y alumnos de la cátedra Introducción a la Investigación (Filosofía y Ciencias de la Educación), realizaron actividades de educación ambiental y prácticas solidarias desde 1999 a 2002, como forma de extensión, transferencia e inserción social de la Universidad. La metodología de investigación-acción participativa optimizó resultados. Otra fuente de datos fueron entrevistas radiales (2001-2003), durante la emisión del Programa del LIPA- Hombre y Ambiente, emitido por Radio Universidad. En la Conclusión estimamos que los antecedentes de EAE revelan que conviene aplicarla en nuestro medio y que la percepción de los problemas de mayor impacto o riesgo ambiental en pobladores del AGC evidencia conocimiento, pero falta mayor compromiso para asumir la propia responsabilidad. Enunciamos fortalezas, debilidades y recomendaciones es por tales antecedentes que las tareas de transferencia y aplicaciones hace "Cuesta Arriba" por lo dificultosa, ya que deben remontarse una serie de inconvenientes algunos de los cuales se han enunciado en la presente comunicación

Cuesta Arriba. Antecedentes epistemológicos y metodológicos para una Evaluación Ambiental Estratégica -EAE- en la ejecución de Políticas Planes y Programas ambientales en el Area del Gran Catamarca -AGC-

En esta comunicación se informa sobre estudios socio ambientales realizados desde las perspectivas epistemológica y metodológica, en el Area del Gran Catamarca. El propósito es contribuir a una Evaluación Estratégica Ambiental. Se examinan antecedentes y se identifica la percepción de la problemática ambiental en diferentes sectores de población.. Para la construcción del Marco teórico se analizaron información escrita (libros, revistas periódicos) y webgrafia e infografia, de origen local, nacional e internacional. El abordaje transversal de la investigación revela la complejidad epistemológica del objeto de estudio cuando se busca alcanzar una visión contemporánea, global y contextualizadora del problema. Complejidad que se evidencia en los resultados de los análisis, de contenidos, multicriterial y hermeneutico. Las indagaciones sobre percepción de la problemática ambiental recogieron testimonios orales, cuestionarios y encuestas cuyas características se describen en el trabajo. Integrantes y alumnos de la cátedra Introducción a la Investigación (Filosofía y Ciencias de la Educación), realizaron actividades de educación ambiental y prácticas solidarias desde 1999 a 2002, como forma de extensión, transferencia e inserción social de la Universidad. La metodología de investigación-acción participativa optimizó resultados. Otra fuente de datos fueron entrevistas radiales (2001-2003), durante la emisión del Programa del LIPA- Hombre y Ambiente, emitido por Radio Universidad. En la Conclusión estimamos que los antecedentes de EAE revelan que conviene aplicarla en nuestro medio y que la percepción de los problemas de mayor impacto o riesgo ambiental en pobladores del AGC evidencia conocimiento, pero falta mayor compromiso para asumir la propia responsabilidad. Enunciamos fortalezas, debilidades y recomendaciones es por tales antecedentes que las tareas de transferencia y aplicaciones hace "Cuesta Arriba" por lo dificultosa, ya que deben remontarse una serie de inconvenientes algunos de los cuales se han enunciado en la presente comunicación

Cuesta Arriba. Antecedentes epistemológicos y metodológicos para una Evaluación Ambiental Estratégica -EAE- en la ejecución de Políticas Planes y Programas ambientales en el Area del Gran Catamarca -AGC-

En esta comunicación se informa sobre estudios socio ambientales realizados desde las perspectivas epistemológica y metodológica, en el Area del Gran Catamarca. El propósito es contribuir a una Evaluación Estratégica Ambiental. Se examinan antecedentes y se identifica la percepción de la problemática ambiental en diferentes sectores de población.. Para la construcción del Marco teórico se analizaron información escrita (libros, revistas periódicos) y webgrafia e infografia, de origen local, nacional e internacional. El abordaje transversal de la investigación revela la complejidad epistemológica del objeto de estudio cuando se busca alcanzar una visión contemporánea, global y contextualizadora del problema. Complejidad que se evidencia en los resultados de los análisis, de contenidos, multicriterial y hermeneutico. Las indagaciones sobre percepción de la problemática ambiental recogieron testimonios orales, cuestionarios y encuestas cuyas características se describen en el trabajo. Integrantes y alumnos de la cátedra Introducción a la Investigación (Filosofía y Ciencias de la Educación), realizaron actividades de educación ambiental y prácticas solidarias desde 1999 a 2002, como forma de extensión, transferencia e inserción social de la Universidad. La metodología de investigación-acción participativa optimizó resultados. Otra fuente de datos fueron entrevistas radiales (2001-2003), durante la emisión del Programa del LIPA- Hombre y Ambiente, emitido por Radio Universidad. En la Conclusión estimamos que los antecedentes de EAE revelan que conviene aplicarla en nuestro medio y que la percepción de los problemas de mayor impacto o riesgo ambiental en pobladores del AGC evidencia conocimiento, pero falta mayor compromiso para asumir la propia responsabilidad. Enunciamos fortalezas, debilidades y recomendaciones es por tales antecedentes que las tareas de transferencia y aplicaciones hace "Cuesta Arriba" por lo dificultosa, ya que deben remontarse una serie de inconvenientes algunos de los cuales se han enunciado en la presente comunicación

Prenatal Vitamin D deficiency induces an early and more severe experimental autoimmune encephalomyelitis in the second generation

In a previous study, we demonstrated that mouse adult F(1) offspring, exposed to a vitamin d deficiency during pregnancy, developed a less severe and delayed Experimental Autoimmune Encephalomyelitis (EAE), when compared with control offspring. We then wondered whether a similar response was observed in the subsequent generation. To answer this question, we assessed F(2) females whose F(1) parents (males or females) were vitamin d-deprived when developing in the uterus of F(0) females. Unexpectedly, we observed that the vitamin d deficiency affecting the F(0) pregnant mice induced a precocious and more severe EAE in the F(2) generation. This paradoxical finding led us to assess its implications for the epidemiology of Multiple Sclerosis (MS) in humans. Using the REFGENSEP database for MS trios (the patient and his/her parents), we collected the parents' dates of birth and assessed a potential season of birth effect that could potentially be indicative of the vitamin d status of the pregnant grandmothers. A trend for a reduced number of births in the Fall for the parents of MS patients was observed but statistical significance was not reached. Further well powered studies are warranted to validate the latter finding.

Gene expression studies in multiple sclerosis

Multiple sclerosis (MS) is a serious neurological disorder affecting young Caucasian individuals, usually with an age of onset at 18 to 40 years old. Females account for approximately 60x of MS cases and the manifestation and course of the disease is highly variable from patient to patient. The disorder is characterised by the development of plaques within the central nervous system (CNS). Many gene expression studies have been undertaken to look at the specific patterns of gene transcript levels in MS. Human tissues and experimental mice were used in these gene-profiling studies and a very valuable and interesting set of data has resulted from these various expression studies. In general, genes showing variable expression include mainly immunological and inflammatory genes, stress and antioxidant genes, as well as metabolic and central nervous system markers. Of particular interest are a number of genes localised to susceptible loci previously shown to be in linkage with MS. However due to the clinical complexity of the disease, the heterogeneity of the tissues used in expression studies, as well as the variable DNA chips/membranes used for the gene profiling, it is difficult to interpret the available information. Although this information is essential for the understanding of the pathogenesis of MS, it is difficult to decipher and define the gene pathways involved in the disorder. Experiments in gene expression profiling in MS have been numerous and lists of candidates are now available for analysis. Researchers have investigated gene expression in peripheral mononuclear white blood cells (PBMCs), in MS animal models Experimental Allergic Encephalomyelitis (EAE) and post mortem MS brain tissues. This review will focus on the results of these studies.

Mimicry of 21. 5 KDa myelin basic protein peptide by a Maedi Visna virus polymerase peptide does not contribute to the pathogenesis of encephalitis in sheep

Epitope mimicry is the theory that an infectious agent such as a virus causes pathological effects via mimicry of host proteins and thus elicits a cross-reactive immune response to host tissues. Weise and Carnegie (1988) found a region of sequence similarity between the pol gene of the Maedi Visna virus (MVV), which induces demyelinating encephalitis in sheep, and myelin basic protein (MBP), which is known to induce experimental allergic encephalitis (EAE) in laboratory animals. In this study, cross-reactions between sera raised in sheep against synthetic peptides of MVV (TGKIPWILLPGR) and 21.5 kDa MBP (SGKVPWLKRPGR) were demonstrated using enzyme-linked immunosorbant assay (ELISA) and thin layer chromatography (TLC) immunoprobing. The antibody responses of MVV-infected sheep were investigated using ELISA against the peptides, and MBP protein, immunoprobing of the peptides on TPC plates and Western blotting against MBP. Slight significant reactions to the 21.5 kDa MBP peptide (P < 0.001) and to a lesser extent sheep MBP (P < 0.004) were detected in ELISA. The MBP peptide evoked stronger responses from more sera than the MVV peptide on immunoprobed TLC plates. On the Western blots, eight of the 23 sheep with Visna had serum reactivity to MBP. This slight reaction to MBP in MVV-infected sheep is of interest because of the immune responses to MBP evident in multiple sclerosis and EAE, but its relevance in Visna is limited since no correlation with disease severity was observed. The cell-mediated immune responses of MVV-infected sheep against similar peptides was assessed. The peptides did not stimulate proliferation of peripheral blood lymphocytes of MVV-infected sheep. Since the MVV peptide was not recognised by antibodies or T lymphocytes from MVV-infected and encephalic sheep, it was concluded that epitope mimicry of this 21.5 kDa MBP peptide by the similar MVV pol peptide was not contributing to the immunopathogenesis of Visna. The slight antibody response to MBP and the MBP peptide can be attributed to by-stander effects of the immunopathology of MVV-induced encephalitis.

Novel Multiple Sclerosis Predisposing Genetic Variants Outside the HLA Region

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS). Both environmental factors and several predisposing genes are required to generate MS. Despite intensive research these risk factors are still largely unknown, the pathogenesis of MS demyelination is poorly understood, and no curative treatment exists. Both prevalence and familial occurrence of MS are exceptionally high in a Finnish population subisolate, Southern Ostrobothnia, presumably due to enrichment of predisposing genetic variants within this region. Previous linkage scan on MS pedigrees from Southern Ostrobothnia detected three main MS loci on chromosomes 5p, 6p (HLA) and 17q. Linkage studies in other populations have also provided independent evidence for the location of MS susceptibility genes in these regions. Further, these loci are syntenic to the experimental autoimmune encephalomyelitis (EAE) susceptibility loci of rodents. In this thesis work an effort was made to localize MS predisposing alleles of the linked loci outside the HLA region by studying familial MS cases from the Southern Ostrobothnia isolate. Analysis of the 5p locus revealed one region, flanking the complement component 7 (C7) gene. The identified relatively rare haplotype seems to have a fairly large effect on genetic susceptibility of MS (frequency MS 12%, controls 4%; p=0.000003, OR=2.73). Evidence for association with alleles of the region and MS was seen also in more heterogeneous populations. Convincingly, plasma C7 protein levels and complement activity correlated with the risk haplotype identified. The finding stimulated us to study other complement cascade genes in MS. No evidence for association could be observed with the complement component coding genes outside 5p. A scan of the 17q locus provided evidence for association with variants of the protein kinase C alpha (PRKCA) gene (p=0.0001). Modest evidence for association with PRKCA was observed also in Canadian MS families. Finally we used a candidate gene based approach to identify potential MS loci. Mutations of DAP12 and TREM2 cause a recessively inherited CNS white matter disease PLOSL. Interestingly, DAP12 and TREM2 are located in MS regions on 6p and 19q, and we tested them as potential candidate genes in the Finnish MS sample. No evidence for association with MS was observed. This thesis provides an example of how extended families from special populations can be utilized in fine-mapping of the linked loci. A first relatively rare MS variant was identified utilizing the strength of a Finnish population subisolate. This variant seems to have an effect on activity of the complement system, which has previously been suggested to have an important role in the pathogenesis of MS.

Increased expression of cystine/glutamate antiporter in multiple sclerosis

Background: Glutamate excitotoxicity contributes to oligodendrocyte and tissue damage in multiple sclerosis (MS). Intriguingly, glutamate level in plasma and cerebrospinal fluid of MS patients is elevated, a feature which may be related to the pathophysiology of this disease. In addition to glutamate transporters, levels of extracellular glutamate are controlled by cystine/glutamate antiporter x(c)(-), an exchanger that provides intracellular cystine for production of glutathione, the major cellular antioxidant. The objective of this study was to analyze the role of the system x(c)(-) in glutamate homeostasis alterations in MS pathology. -- Methods: Primary cultures of human monocytes and the cell line U-937 were used to investigate the mechanism of glutamate release. Expression of cystine glutamate exchanger (xCT) was quantified by quantitative PCR, Western blot, flow cytometry and immunohistochemistry in monocytes in vitro, in animals with experimental autoimmune encephalomyelitis (EAE), the animal model of MS, and in samples of MS patients. -- Results and discussion: We show here that human activated monocytes release glutamate through cystine/glutamate antiporter x(c)(-) and that the expression of the catalytic subunit xCT is upregulated as a consequence of monocyte activation. In addition, xCT expression is also increased in EAE and in the disease proper. In the later, high expression of xCT occurs both in the central nervous system (CNS) and in peripheral blood cells. In particular, cells from monocyte-macrophage-microglia lineage have higher xCT expression in MS and in EAE, indicating that immune activation upregulates xCT levels, which may result in higher glutamate release and contribution to excitotoxic damage to oligodendrocytes. -- Conclusions: Together, these results reveal that increased expression of the cystine/glutamate antiporter system x(c)(-) in MS provides a link between inflammation and excitotoxicity in demyelinating diseases.