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1.
J Comp Pathol ; 148(4): 335-44, 2013 May.
Article in English | MEDLINE | ID: mdl-23063106

ABSTRACT

Chronic osteoarthritis (OA) is a degenerative disease of the articular cartilage. DNA-binding high mobility group protein B1 (HMGB1) is released on cellular death/activation and acts as an endogenous danger signal and a proinflammatory cytokine. Matrix metalloproteinase (MMP)-2 and in MMP-9 are induced to mediate proteolytic degradation/remodelling of joint tissues. Collagen degradation in the bone and synovium leads to release of type I collagen-derived cross-linked carboxy-terminal telopeptide (ICTP). These molecules have been linked to the pathogenesis of OA and could have potential as synovial fluid (SF) biomarkers in OA. Cartilage and SF were obtained from 27 dairy bulls (30-61 months old) and control cartilage from six young healthy dairy bulls. OA lesions were evaluated grossly (five grades), histologically (seven Osteoarthritis Research Society International [ORSI] grades) and immunohistochemically (four HMGB1 grades). The OARSI lesion score was calculated as the product of the OARSI grade and the OARSI score (the total area of the lesions). SF concentrations of HMGB1, MMP-2 and -9 and ICTP were measured by enzyme-linked immunosorbent assay, gelatin zymography and radioimmunoassay, respectively. Seventy-two percent (39/54) of stifle joints and 85% (23/27) of the dairy bulls had at least one gross OA lesion and 94% of the lesions were localized to the distal end of the femur, with the patellar groove and the lateral trochlear ridge being predilection sites. Gross and histological grades correlated with the HMGB1 grade, but SF total cell count, percent neutrophils or the measured biomarkers did not correlate with the tissue lesions, with the exception of ICTP concentration, which correlated with the total joint score. The switch of HMGB1 from DNA-binding nuclear protein to an extracellular alarmin/cytokine correlates with the gross and histological grades of OA tissue lesions. However, the activity and extent of the tissue lesions did not correlate with other SF biomarkers, perhaps because the histological grades represent outcome measures, while SF reflects process parameters. The only exception was ICTP concentration, which reflects enhanced destruction/remodelling.


Subject(s)
Cartilage, Articular/metabolism , Knee Joint/metabolism , Osteoarthritis/veterinary , Synovial Fluid/metabolism , Animals , Biomarkers/metabolism , Cartilage, Articular/pathology , Cattle , Collagen Type I/metabolism , Femur/metabolism , Femur/pathology , HMGB1 Protein/metabolism , Knee Joint/pathology , Male , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Osteoarthritis/metabolism , Osteoarthritis/pathology , Peptides/metabolism
2.
Theriogenology ; 77(1): 201-5, 2012 Jan 01.
Article in English | MEDLINE | ID: mdl-21924472

ABSTRACT

Multiple ovulation embryo transfer (MOET) is used to make more rapid progress in animal breeding schemes. On dairy farms, where female calves are more desired, embryo sex diagnosis is often performed before embryo transfer. Fresh transfers have been favored after biopsy due to cumulative drop in pregnancy rates following cryopreservation. The aim of this study was to explore whether exposure to ascorbic acid (AC) during biopsy and freezing increases the viability of biopsied embryos after cryopreservation. Data on presumptive pregnancy and calving rates of biopsied and cryopreserved/overnight-cultured embryos were gathered. Results showed differences in presumptive pregnancy rates between the groups: 45% for both biopsied-cryopreserved groups (control and AC), 51% for biopsied-overnight-cultured embryos and 80% for intact-fresh embryos. Differences between the groups were also apparent in calving rates: 22% for biopsied-cryopreserved control embryos, 31% for biopsied-cryopreserved AC-embryos, 23% for biopsied-overnight-cultured embryos and 63% for intact-fresh embryos. It is concluded that manipulated embryos are associated with lower presumptive pregnancy and calving rates compared with intact-fresh embryos. The highest calving rates for groups of manipulated embryos were achieved in the AC-group. Therefore, addition of AC can be recommended if biopsy is combined with freezing before transfer.


Subject(s)
Ascorbic Acid/pharmacology , Cattle/embryology , Cryopreservation/veterinary , Cryoprotective Agents/pharmacology , Embryo, Mammalian/drug effects , Animals , Cryopreservation/methods , Embryo Transfer/veterinary , Female , Male , Pregnancy , Pregnancy Rate , Sex Determination Analysis/veterinary
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