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Biomedical subjects

A Goodship

Publications and source records attributed to A Goodship.

9 recordsLinked to original sources

Cytokine induced metalloproteinase expression and activity does not correlate with focal susceptibility of articular cartilage to degeneration.

OBJECTIVE: To determine whether the focal susceptibility to cartilage degeneration in joints is related to a differential response to cytokine stimulation. METHODS: Compare aggrecan and collagen catabolism in in-vitro models of cartilage degradation induced by retinoic acid (RA), interleukin-1 (IL-1), tumor necrosis factor alpha (TNF) and IL-1 plus oncostatin M (OSM). Glycosaminoglycan (GAG) and hydroxyproline (HyPro) quantification and Western immunoblot analyses of aggrecan and collagen degradation products were undertaken in explant cultures of normal cartilage from regions of equine joints with a known high and low susceptibility to degeneration in disease. RNA isolation and semi quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis were performed to determine the expression of aggrecanases, matrix metalloproteinases (MMPs) and their inhibitors. RESULTS: Although the rate of basal cartilage aggrecan turnover was dependent on joint region there was no difference in the response of different cartilages to cytokines. Individual animals did show a significant difference in the response of certain cartilages to cytokines, with both decreased and increased aggrecan loss in cartilage with a low susceptibility to degeneration. Aggrecan release in both short- and long-term cultures from all cartilages was associated with increased cleavage by aggrecanases rather than MMPs. There was a poor correlation between expression of aggrecanases, MMPs or their inhibitors and cytokine induced aggrecan catabolism. IL-1 alone was able to stimulate collagen breakdown in equine articular cartilage and surprisingly, significantly more collagen loss was induced in cartilage from regions less susceptible to degeneration. CONCLUSIONS: Collectively, these studies suggest that a regional difference in response to catabolic cytokines is unlikely to be a factor in the initiation of focal cartilage degeneration in osteoarthritis (OA).

Aggrecans↗

The effects of hyaluronic acid on articular chondrocytes.

The purpose of this study was to examine the effects of hyaluronic acid supplementation on chondrocyte metabolism in vitro. The clinical benefits of intra-articular hyaluronic acid injections are thought to occur through improved joint lubrication. Recent findings have shown that exogenous hyaluronic acid is incorporated into articular cartilage where it may have a direct biological effect on chondrocytes through CD44 receptors.Bovine articular chondrocytes were isolated and seeded into alginate constructs. These were cultured in medium containing hyaluronic acid at varying concentrations. Samples were assayed for biochemical and histological changes. There was a dose-dependent response to the exposure of hyaluronic acid to bovine articular chondrocytes in vitro. Low concentrations of hyaluronic acid (0.1 mg/mL and 1 mg/mL) significantly increase DNA, sulphated glycosaminoglycan and hydroxyproline synthesis. Immunohistology confirmed the maintenance of cell phenotype with increased matrix deposition of chondroitin-6-sulphate and collagen type II. These findings confirm a stimulatory effect of hyaluronic acid on chondrocyte metabolism.

Animals↗

Numerical simulations of subsurface probing in diffusely scattering media using spatially offset Raman spectroscopy.

We present the first elementary model predicting how Raman intensities vary for a range of experimental variables for spatially offset Raman spectroscopy (SORS), a recently proposed technique for the effective retrieval of Raman spectra of subsurface layers in diffusely scattering media. The model was able to reproduce the key observations made from the first SORS experiments, namely the dependence of Raman signal intensities on the spatial offset between the illumination and collection points and the relative contributions to the overall spectrum from the top layer and sub-layer. The application of the SORS concept to a three-layer system is also discussed. The model also clearly indicates that an annular geometry, rather than a point-collection geometry, which was used in the earlier experiments, would yield much improved data.

Algorithms↗

Gene expression array profile of human osteosarcoma.

The aims of this pilot study were to determine whether needle and open biopsies from osteosarcoma (OS) provide sufficient quality of mRNA for cDNA array analyses to gain insights into the expression profile of OS. A total of 22 samples collected from OS were used for array analyses. A primary cell culture was also established from one of the OS biopsies. Total RNA was extracted and probes were generated for cDNA arrays. cDNA probes were made for all the 22 samples. Two of these samples were needle core bone biopsies. Statistical analysis confirmed the reliability of array data obtained in 16 of the 22 samples. Known genes involved in bone metabolism and osteosarcoma were identified as highly expressed, and the putative new marker Ezrin was also identified. Confirmatory immunohistochemical staining using the Ezrin antibody was performed in a selection of samples.

Adolescent↗

The distribution of cartilage oligomeric matrix protein (COMP) in equine carpal articular cartilage and its variation with exercise and cartilage deterioration.

Based on previous studies where tendons receiving the most load have been shown to have the highest levels of cartilage oligomeric matrix protein (COMP), we hypothesized that COMP distribution in articular cartilage may be influenced by mechanical loading. This investigation aimed (a) to describe the pattern of COMP immunoreactivity in middle carpal joint cartilage of two-year-old Thoroughbred horses; (b) to determine topographical variations; (c) to compare high (group 1) and low (group 2) intensity training and (d) to describe COMP immunoreactivity at sites with early osteoarthritis. Group 1 (n =6) underwent a 19 week high-intensity treadmill training programme and group 2 (n =6) were given daily walking until euthanasia. Dorsal and palmar sites on radial and third carpal articular surfaces were prepared. Immunohistochemistry was performed with polyclonal rabbit anti-equine COMP antiserum using a biotin-streptavidin/peroxidase method. Results showed: (a) intracellular immunoreactivity was present in all cartilage zones, but the distribution of COMP staining within the matrix varied between cartilage zones; (b) differences in distribution between sites were not observed, but total COMP levels in exercised horses (n =2) did vary between sites with dorsal sites containing less COMP than palmar sites on the radial, intermediate and third carpal lateral facet; (c) group 1 cartilage showed marked interterritorial distribution in the deep layer compared to group 2 where staining was more generalized throughout the matrix and (d) fibrillated cartilage showed increased local immunoreactivity in the matrix. These findings demonstrate zonal variations in equine COMP distribution which may be influenced by loading.

Animals↗

Investigation of bone changes in microgravity during long and short duration space flight: comparison of techniques.

BACKGROUND: Loss of bone mass is a continuing problem in long-term space flight. Although counter-measure programmes have been developed, effective assessment of these programmes is hampered by a lack of monitoring techniques that can be used in-flight. MATERIALS AND METHODS: Three techniques were used to evaluate changes in bone during two missions of 180 and 20 days to the MIR space station, involving three subjects. Dual energy X-ray absorptiometry (DXA) was used before and after flight to measure whole body and regional bone mineral density (BMD). Ultrasonic measurements of velocity (SOS) and broadband attenuation (BUA) of the calcaneus were measured during the 180 day mission and before and after the 20 day mission. Phase velocity of flexural waves in the tibia was also measured on the same days as the ultrasonic measurements of the calcaneus. RESULTS: DXA measurements demonstrated significant variation between different sites in the body for changes in BMD, with the greatest changes occurring in the lumbar spine and proximal femur. There was a trend for increasing phase velocity in the tibia during the 180 mission, but this was not significant. BUA and SOS measurements of the calcaneus showed consistent but divergent patterns of changes during the mission. CONCLUSION: Although in-flight measurements of bone using ultrasound or phase velocity may provide information on the kinetics of bone loss in space flight, the heterogeneity of response in the skeleton means that it is difficult to predict overall bone loss from measurements at one particular site.

Absorptiometry, Photon↗

Equine carpal articular cartilage fibronectin distribution associated with training, joint location and cartilage deterioration.

Processes involved in equine carpal osteochondral injury have not been established. In other species, fibronectin appears important in chondrocyte-matrix interactions, and levels are increased in osteoarthritis. This investigation aimed to (a) describe fibronectin immunoreactivity in the middle carpal joint of 2-year-old Thoroughbreds, (b) determine topographical variations, (c) compare strenuously trained (Group 1) or gently exercised horses (Group 2) and (d) describe sites with early osteoarthritis. Group 1 (n = 6) underwent a 19 week high intensity treadmill training programme. Group 2 (n = 6) underwent 40 min walking until euthanasia. Dorsal and palmar sites on radial, intermediate and third carpal articular surfaces were prepared. Immunohistochemistry was performed using a biotin-streptavidin/peroxidase method. Cross-reactivity of rabbit antihuman fibronectin antiserum with equine fibronectin was confirmed using Western blotting. Results showed: (a) fibronectin was present primarily in pericellular and interterritorial matrix locations, (b) dorsal sites had zonal immunoreactivity compared to palmar sites, (c) Group 1 dorsal radial carpal cartilage had increased superficial staining compared to Group 2 and (d) fibrillated cartilage showed increased intracellular and local matrical immunoreactivity (superficial zone). These findings suggest topographical and exercise-related variations in fibronectin distribution, and indicate equine fibronectin is localised at sites of cartilage degeneration and released into the matrix by chondrocytes in the local area.

Animals↗

Bone stress in the horse forelimb during locomotion at different gaits: a comparison of two experimental methods.

Longitudinal stresses acting in the cranial and caudal cortices of the radius and the dorsal and palmar cortices of the metacarpus in the horse were determined using two independent methods simultaneously. One approach involved the use of rosette strain gauges to record in vivo bone strain; the other involved filming the position of the horse's forelimb as it passed over a force plate. Agreement between the two analyses was better for the radius than for the metacarpus. Both methods showed the radius to be loaded primarily in sagittal bending, acting to place the caudal cortex in compression and the cranial cortex in tension. At each gait the magnitude of peak stress in each cortex based on the film/force analysis was 1.5-2 times higher than that determined from the bone strain recordings. In the metacarpus, the magnitude of stress in each cortex calculated from the film/force method was 2-3 times greater at each gait than that shown by the bone strain recordings. However, whereas the film/force analysis indicated that the metacarpus was loaded in sagittal bending (acting to place the palmar cortex in compression and the dorsal cortex in tension), the bone strain recordings showed the metacarpus to be loaded primarily in axial compression at each gait. Because the film/force method depends on an accurate measure of limb segment orientation relative to the direction of ground reaction force, comparatively small errors in calculations of bending moments may lead to a significant difference in the level and distribution of stress determined to act in the bone's cortices. The discrepancy in metacarpal loading obtained by the two methods may be explained in part by the simplicity of the biomechanical model which, for instance, neglected the force exerted by the sesamoids on the distal end of the metacarpus. The records of stress determined from the in vivo bone strain recordings showed that each bone was subjected to a consistent loading regime despite changes of gait. Such a consistent strain distribution should allow these bones to maximize economy in the use of tissue required to support the dynamic loads applied. Peak stresses measured from the bone strain recordings in the radius during locomotion at constant speed (-40.8 +/- 4.1 MN m-2) were significantly larger than those in the metacarpus (-25.1 +/- 2.8 MN m-2), regardless of speed and gait. During acceleration and deceleration, however, peak stress rose dramatically in the metacarpus (-40.6 +/- 3.4 MN m-2) but remained constant in the radius (-37.8 +/- 5.8 MN m-2).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

ATP loss with exercise in muscle fibres of the gluteus medius of the thoroughbred horse.

Muscle ATP loss with exercise has implications both to the causes of fatigue and muscle damage. To study this at the single muscle fibre level, five trained thoroughbred horses performed consecutive 90 second gallops on an inclined treadmill followed by a final gallop to fatigue. Biopsies of the m. gluteus medius were taken at rest, post-exercise and during 24 hour recovery. Blood lactate was 20.0 mmol litre-1 or more, and plasma NH3 300-800 mumol litre-1, following the final gallop. Minimal changes occurred in the plasma markers, CK and AST. ATP loss with exercise was 32.2 (SD 12.2) per cent. Following exercise single fibre ATP contents showed a much broader distribution than at rest, with contents in some close to zero. Following five and 24 hour recovery, however, frequency distribution curves were close to those seen at rest. There was no difference in the ATP contents of types I, IIa and IIb at rest of with exercise or recovery. The results pointed to marked heterogeneity between individual fibres in their biochemical response with exercise, independent of fibre type.

Adenosine Triphosphate↗