| 1. |
- Blomström, Anne-Lie, et al.
(författare)
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Astrovirus as a possible cause of congenital tremor type AII in piglets?
- 2014
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Ingår i: Acta Veterinaria Scandinavica. - : Springer Science and Business Media LLC. - 0044-605X .- 1751-0147. ; 56
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Tidskriftsartikel (refereegranskat)abstract
- Background: Congenital tremor is associated with demyelination of the brain and spinal cord and is clinically noted as outbreaks of trembling and shaking in newborn piglets during a limited time-period. Six forms of the disease have been described, where form AII may be caused by an, as yet, unidentified viral infection. This study aimed to investigate the presence of astrovirus and circovirus by sequencing and polymerase chain reaction (PCR) analysis and by relating the findings to the occurrence of disease and lesions in the brain, in 4-6 days-old piglets obtained from a clinical outbreak of congenital tremor.Results: In piglets with congenital tremor, there were mild to moderate vacuolar changes of the white matter in the cerebrum, brain stem and cerebellum. In healthy piglets, less conspicuous vacuolar changes were detected. One healthy and one diseased piglet were positive for porcine circovirus type 2. The nested pan-PCR showed the presence of astrovirus in at least one brain region in all piglets and by sequencing, two different porcine astrovirus lineages were identified.Conclusions: The results do not support previous studies identifying porcine circovirus type 2 as the cause of congenital tremor. The demonstration of astrovirus in the brain of piglets suffering from congenital tremor is interesting. However, astrovirus was demonstrated in both healthy and diseased individuals and therefore, further studies are warranted to determine the possible involvement of astrovirus in the pathogenesis of congenital tremor in pigs.
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| 2. |
- Ley, Cecilia, et al.
(författare)
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Effects of high mobility group box protein-1, interleukin-1 beta, and interleukin-6 on cartilage matrix metabolism in three-dimensional equine chondrocyte cultures
- 2011
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Ingår i: Connective Tissue Research. - : Informa UK Limited. - 0300-8207 .- 1607-8438. ; 52, s. 290-300
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Tidskriftsartikel (refereegranskat)abstract
- The effects of high mobility group box protein (HMGB)-1, interleukin (IL)-1 beta, and IL-6 on equine articular chondrocytes were investigated, with emphasis on detecting differences between anatomical sites exposed to different loading in vivo, using three-dimensional (3D) cell cultures established with chondrocytes from dorsal radial facet (DRF, highly loaded) and palmar condyle (PC, less loaded) of the third carpal bone (C3). Expression of important genes involved in cartilage metabolism, presence of glycosaminoglycans and cartilage oligomeric matrix protein (COMP) in pellets, and concentrations of matrix metalloproteinase (MMP)-13 and aggrecan epitope CS 846 were evaluated. Compared to controls, IL-1 beta treatment increased gene expression of versican, matrix-degrading enzymes, and tissue inhibitor of metalloproteinase (TIMP)-1, and decreased aggrecan and collagen type I and type II expression. In addition, IL-1 beta-treated pellets showed decreased safranin O staining and increased COMP immunostaining and MMP-13 concentrations in culture supernatants. Effects of IL-6 and HMGB-1 on gene expression were variable, although upregulation of Sry-related high-mobility group box 9 (Sox9) was often present and statistically increased in HMGB-1-treated pellets. Response to cytokines rarely differed between DRF and PC pellets. Thus, site-associated cartilage deterioration in equine carpal osteoarthritis (OA) is not explained by topographically different responses to inflammatory mediators. Differences in gene expressions of structural matrix proteins in untreated DRF and PC pellets were noted in the youngest horses, which may indicate differences in the chondrocytes potential to produce matrix in vivo. Overall, a strong catabolic response was induced by IL-1 beta, whereas slight anabolic effects were induced by IL-6 and HMGB-1.
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| 3. |
- Ley, Cecilia, et al.
(författare)
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Effects of high mobility group box protein-1, interleukin-1β, and interleukin-6 on cartilage matrix metabolism in three-dimensional equine chondrocyte cultures.
- 2011
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Ingår i: Connective tissue research. - : Informa UK Limited. - 1607-8438 .- 0300-8207. ; 52:4, s. 290-300
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Tidskriftsartikel (refereegranskat)abstract
- The effects of high mobility group box protein (HMGB)-1, interleukin (IL)-1β, and IL-6 on equine articular chondrocytes were investigated, with emphasis on detecting differences between anatomical sites exposed to different loading in vivo, using three-dimensional (3D) cell cultures established with chondrocytes from dorsal radial facet (DRF, highly loaded) and palmar condyle (PC, less loaded) of the third carpal bone (C3). Expression of important genes involved in cartilage metabolism, presence of glycosaminoglycans and cartilage oligomeric matrix protein (COMP) in pellets, and concentrations of matrix metalloproteinase (MMP)-13 and aggrecan epitope CS 846 were evaluated. Compared to controls, IL-1β treatment increased gene expression of versican, matrix-degrading enzymes, and tissue inhibitor of metalloproteinase (TIMP)-1, and decreased aggrecan and collagen type I and type II expression. In addition, IL-1β-treated pellets showed decreased safranin O staining and increased COMP immunostaining and MMP-13 concentrations in culture supernatants. Effects of IL-6 and HMGB-1 on gene expression were variable, although upregulation of Sry-related high-mobility group box 9 (Sox9) was often present and statistically increased in HMGB-1-treated pellets. Response to cytokines rarely differed between DRF and PC pellets. Thus, site-associated cartilage deterioration in equine carpal osteoarthritis (OA) is not explained by topographically different responses to inflammatory mediators. Differences in gene expressions of structural matrix proteins in untreated DRF and PC pellets were noted in the youngest horses, which may indicate differences in the chondrocytes potential to produce matrix in vivo. Overall, a strong catabolic response was induced by IL-1β, whereas slight anabolic effects were induced by IL-6 and HMGB-1.
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| 4. |
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| 5. |
- Ley, Cecilia
(författare)
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Inflammatory response in equine joints : studies on proinflammatory cytokines in diseased joints and chondrocyte cultures
- 2010
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Proinflammatory cytokines mediate inflammatory responses as well as regulate tissue metabolism. Thus, they may provide a link between inflammation and other pathologic findings seen in equine joint disease. The aims of this thesis were to gain a deeper understanding of the development of chondral pathology in equine osteoarthritis (OA) by obtaining increased knowledge of inflammatory processes in the joint, and to investigate proinflammatory cytokines as markers of joint pathology. Measurements of interleukin (IL)-6 and tumour necrosis factor (TNF) activities in synovial fluid from clinical cases of carpal joint disease revealed increased concentrations of bioactive IL-6 in joints with osteochondral fragments (OCF). Immunohistochemical staining for IL-6 and high mobility group box protein (HMGB)-1 in tissues from diseased carpal and fetlock joints identified both OCF and synovial membranes as cellular sources of IL-6 and extracellular/cytoplasmic HMBG-1. Histological assessment of synovial membrane biopsies from immunostained joints showed increased grade of synovitis in diseased compared to healthy joints. The effects of IL-6, IL-1β and HMGB-1 on chondrocyte metabolism was studied in vitro, and focused on gene expression analyses. A catabolic response to IL-1β was detected, whereas the response to IL-6 and HMGB-1 was varied but indicated promotion of cartilage formation. Analyses of the ultrastructural immunolocalisation of HMGB-1 in IL-1β and IL-6 treated pellets, showed a trend for decreased nuclear/cytoplasmic ratio, and increased extracellular matrix density of HMGB-1 after IL-6 treatment. Theses studies gained insight to presence of TNF, IL-6 and HMGB-1 in equine joints, and indicated an increased inflammatory response in joints with OCF. The difference in metabolic response of chondrocytes to IL-1β versus IL-6 or HMGB-1 in vitro, and differences in HMGB-1 translocation after IL-1β and IL-6 treatment suggest that the outcome of cartilage pathology in equine joint disease may relate to the individual contributions of cytokine activity and cytokine interactions.
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| 6. |
- Löfgren, Maria, et al.
(författare)
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Cell and matrix modulation in prenatal and postnatal equine growth cartilage, zones of Ranvier and articular cartilage
- 2014
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Ingår i: Journal of Anatomy. - : Wiley. - 0021-8782 .- 1469-7580. ; 225:5, s. 548-568
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Tidskriftsartikel (refereegranskat)abstract
- Formation of synovial joints includes phenotypic changes of the chondrocytes and the organisation of their extracellular matrix is regulated by different factors and signalling pathways. Increased knowledge of the normal processes involved in joint development may be used to identify similar regulatory mechanisms during pathological conditions in the joint. Samples of the distal radius were collected from prenatal and postnatal equine growth plates, zones of Ranvier and articular cartilage with the aim of identifying Notch signalling components and cells with stem cell-like characteristics and to follow changes in matrix protein localisation during joint development. The localisation of the Notch signalling components Notch1, Delta4, Hes1, Notch dysregulating protein epidermal growth factor-like domain 7 (EGFL7), the stem cell-indicating factor Stro-1 and the matrix molecules cartilage oligomeric matrix protein (COMP), fibromodulin, matrilin-1 and chondroadherin were studied using immunohistochemistry. Spatial changes in protein localisations during cartilage maturation were observed for Notch signalling components and matrix molecules, with increased pericellular localisation indicating new synthesis and involvement of these proteins in the formation of the joint. However, it was not possible to characterise the phenotype of the chondrocytes based on their surrounding matrix during normal chondrogenesis. The zone of Ranvier was identified in all horses and characterised as an area expressing Stro-1, EGFL7 and chondroadherin with an absence of COMP and Notch signalling. Stro-1 was also present in cells close to the perichondrium, in the articular cartilage and in the fetal resting zone, indicating stem cell-like characteristics of these cells. The presence of stem cells in the articular cartilage will be of importance for the repair of damaged cartilage. Perivascular chondrocytes and hypertrophic cells of the cartilage bone interface displayed positive staining for EGFL7, which is a novel finding and suggests a role of EGFL7 in the vascular infiltration of growth cartilage.
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| 7. |
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| 8. |
- Svala, Emilia, et al.
(författare)
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Effects of interleukin-6 and interleukin-1 beta on expression of growth differentiation factor-5 and Wnt signaling pathway genes in equine chondrocytes
- 2014
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Ingår i: American Journal of Veterinary Research. - : American Veterinary Medical Association (AVMA). - 0002-9645 .- 1943-5681. ; 75:2, s. 132-140
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Tidskriftsartikel (refereegranskat)abstract
- Objective-To determine the effects of interleukin (IL)-6 and IL-1 beta stimulation on expression of growth differentiation factor (GDF)-5 and Wnt signaling pathway genes in equine chondrocytes. Sample-Macroscopically normal articular cartilage samples from 6 horses and osteochondral fragments (OCFs) from 3 horses. Procedures-Chondrocyte pellets were prepared and cultured without stimulation or following stimulation with IL-6 or IL-1 beta for 1, 2, 12, and 48 hours; expression of GDF-5 was determined with a quantitative real-time PCR assay. Expression of genes in various signaling pathways was determined with microarrays for pellets stimulated for 1 and 2 hours. Immunohistochemical analysis was used to detect GDF-5, glycogen synthase kinase 3 beta (GSK-3 beta), and beta-catenin proteins in macroscopically normal cartilage samples and OCFs. Results-Chondrocytes stimulated with IL-6 had significantly higher GDF-5 expression within 2 hours versus unstimulated chondrocytes. Microarray analysis of Wnt signaling pathway genes indicated expression of GSK-3 beta and coiled-coil domain containing 88C increased after 1 hour and expression of beta-catenin decreased after 2 hours of IL-6 stimulation. Results of immunohistochemical detection of proteins were similar to microarray analysis results. Chondrocytes in macroscopically normal articular cartilage and OCFs had immunostaining for GDF-5. Conclusion and Clinical Relevance-Results indicated IL-6 stimulation decreased chondrocyte expression of the canonical Wnt signaling pathway transactivator beta-catenin, induced expression of inhibitors of the Wnt pathway, and increased expression of GDF-5. This suggested IL-6 may inhibit the Wnt signaling pathway with subsequent upregulation of GDF-5 expression. Anabolic extracellular matrix metabolism in OCFs may be attributable to GDF-5 expression. This information could be useful for development of cartilage repair methods.
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