Effect of circadian rhythm, age, training and acute lameness on serum concentrations of cartilage oligomeric matrix protein (COMP) neo-epitope in horses.

BACKGROUND: Molecular serum markers that can identify early reversible osteoarthritis (OA) in horses are lacking. OBJECTIVES: We studied serum concentrations of a novel cartilage oligomeric matrix protein (COMP) neo-epitope in horses subjected to short-term exercise and with acute lameness. The effects of circadian rhythm and age were also evaluated. STUDY DESIGN: Longitudinal studies in healthy horses and cross-sectional comparison of lame and non-lame horses. METHODS: Sera were collected from five horses before and after short-term interval exercise and during full-day box rest. Sera from 32 acutely lame horses were used to evaluate age-related effects. Independent samples from control horses (n = 41) and horses with acute lameness (n = 71) were included. COMP neo-epitope concentrations were analysed using custom-developed inhibition ELISAs validated for equine serum. The presence of COMP neo-epitope was delineated in healthy and osteoarthritic articular cartilage with immunohistochemistry. RESULTS: COMP neo-epitope concentrations decreased after speed training but returned to baseline levels post-exercise. No correlations between age and serum COMP neo-epitope concentrations were found (r = 0.0013). The mean (±s.d.) serum concentration of COMP neo-epitope in independent samples from non-lame horses was 0.84 ± 0.38 µg/mL, and for lame horses was 5.24 ± 1.83 µg/mL (P<0.001). Antibodies against COMP neo-epitope did not stain normal articular cartilage, but intracytoplasmic staining was found in superficial chondrocytes of mild OA cartilage and in the extracellular matrix of moderately osteoarthritic cartilage. MAIN LIMITATIONS: ELISA was based on polyclonal antisera rather than a monoclonal antibody. There is a sex and breed bias within the groups of horses, also it could have been of value to include horses with septic arthritis and tendonitis and investigated joint differences. CONCLUSIONS: This COMP neo-epitope can be measured in sera, and results indicate that it could be a biomarker for pathologic fragmentation of cartilage in connection with acute joint lameness.

Precision of bone density and micro-architectural properties at the distal radius and tibia in children: an HR-pQCT study.

Precision errors need to be known when monitoring bone micro-architecture in children with HR-pQCT. Precision errors for trabecular bone micro-architecture ranged from 1 to 8% when using the standard evaluation at the radius and tibia. Precision errors for cortical bone micro-architecture ranged from 1 to 11% when using the advanced cortical evaluation. INTRODUCTION: Our objective was to define HR-pQCT precision errors (CV%RMS) and least significant changes (LSCs) at the distal radius and tibia in children using the standard evaluation and the advanced cortical evaluation. METHODS: We scanned the distal radius (7% of ulnar length) and tibia (8% of tibia length) of 32 children (age range 8-13; mean age 11.3; SD 1.6 years) twice (1 week apart) using HR-pQCT (XtremeCT1). We calculated root-mean-squared coefficients of variation (CV%RMS) to define precision errors and LSC to identify differences required to detect change. RESULTS: Precision errors ranged between 1-8 and 1-5% for trabecular bone outcomes (obtained with standard evaluation) and between 1.5-11 and 0.5-6% for cortical bone outcomes (obtained with advanced cortical evaluation) at the distal radius and tibia, respectively. Related LSCs ranged between 3-21 and 3-14% for trabecular bone outcomes and between 4-30 and 2-16% for cortical bone outcomes at the distal radius and tibia, respectively. CONCLUSIONS: HR-pQCT precision errors were between 1 and 8% (LSC 3-21%) for trabecular bone outcomes and 1 and 11% (LSC 2-30%) for cortical bone outcomes at the radius and tibia in children. Cortical bone outcomes obtained using the advanced cortical evaluation appeared to have lower precision errors than cortical outcomes derived using the standard evaluation. These findings, combined with better-defined cortical bone contours with advanced cortical evaluation, indicate that metrics from advanced cortical evaluation should be utilized when monitoring cortical bone properties in children.

Precision of pQCT-measured total, trabecular and cortical bone area, content, density and estimated bone strength in children.

OBJECTIVES: To define pQCT precision errors, least-significant-changes, and identify associated factors for bone outcomes at the radius and tibia in children. METHODS: We obtained duplicate radius and tibia pQCT scans from 35 children (8-14yrs). We report root-mean-squared coefficient of variation (CV%RMS) and 95% limits-of-agreement to characterize repeatability across scan quality and least-significant-changes for bone outcomes at distal (total and trabecular area, content and density; and compressive bone strength) and shaft sites (total area and content; cortical area content, density and thickness; and torsional bone strength). We used Spearman's rho to identify associations between CV% and time between measurements, child's age or anthropometrics. RESULTS: After excluding unanalyzable scans (6-10% of scans per bone site), CV%RMS ranged from 4% (total density) to 19% (trabecular content) at the distal radius, 4% (cortical content) to 8% (cortical thickness) at the radius shaft, 2% (total density) to 14% (trabecular content) at the distal tibia and from 2% (cortical content) to 6% (bone strength) at the tibia shaft. Precision errors were within 95% limits-of-agreement across scan quality. Age was associated (rho -0.4 to -0.5, p⟨0.05) with CV% at the tibia. CONCLUSION: Bone density outcomes and cortical bone properties appeared most precise (CV%RMS⟨5%) in children.

Cartilage oligomeric matrix protein neoepitope in the synovial fluid of horses with acute lameness: A new biomarker for the early stages of osteoarthritis.

BACKGROUND: Clinical tools to diagnose the early changes of osteoarthritis (OA) that occur in the articular cartilage are lacking. OBJECTIVES: We sought to identify and quantify a novel cartilage oligomeric matrix protein (COMP) neoepitope in the synovial fluid from the joints of healthy horses and those with different stages of OA. STUDY DESIGN: In vitro quantitative proteomics and assay development with application in synovial fluids samples obtained from biobanks of well-characterised horses. METHODS: Articular cartilage explants were incubated with or without interleukin-1ß for 25 days. Media were analysed via quantitative proteomics. Synovial fluid was obtained from either normal joints (n = 15) or joints causing lameness (n = 17) or with structural OA lesions (n = 7) and analysed for concentrations of the COMP neoepitope using a custom-developed inhibition enzyme-linked immunosorbent assay (ELISA). Explants were immunostained with polyclonal antibodies against COMP and the COMP neoepitopes. RESULTS: Semitryptic COMP peptides were identified and quantified in cell culture media from cartilage explants. A rabbit polyclonal antibody was raised against the neoepitope of the N-terminal portion of one COMP fragment (sequence SGPTHEGVC). An inhibition ELISA was developed to quantify the COMP neoepitope in synovial fluid. The mean concentration of the COMP neoepitope significantly increased in the synovial fluid from the joints responsible for acute lameness compared with normal joints and the joints of chronically lame horses and in joints with chronic structural OA. Immunolabelling for the COMP neoepitope revealed a pericellular staining in the interleukin-1ß-stimulated explants. MAIN LIMITATIONS: The ELISA is based on polyclonal antisera rather than a monoclonal antibody. CONCLUSIONS: The increase in the COMP neoepitope in the synovial fluid from horses with acute lameness suggests that this neoepitope has the potential to be a unique candidate biomarker for the early molecular changes in articular cartilage associated with OA.

Amazon River enhances diazotrophy and carbon sequestration in the tropical North Atlantic Ocean.

The fresh water discharged by large rivers such as the Amazon is transported hundreds to thousands of kilometers away from the coast by surface plumes. The nutrients delivered by these river plumes contribute to enhanced primary production in the ocean, and the sinking flux of this new production results in carbon sequestration. Here, we report that the Amazon River plume supports N(2) fixation far from the mouth and provides important pathways for sequestration of atmospheric CO(2) in the western tropical North Atlantic (WTNA). We calculate that the sinking of carbon fixed by diazotrophs in the plume sequesters 1.7 Tmol of C annually, in addition to the sequestration of 0.6 Tmol of C yr(-1) of the new production supported by NO(3) delivered by the river. These processes revise our current understanding that the tropical North Atlantic is a source of 2.5 Tmol of C to the atmosphere [Mikaloff-Fletcher SE, et al. (2007) Inverse estimates of the oceanic sources and sinks of natural CO(2) and the implied oceanic carbon transport. Global Biogeochem Cycles 21, doi:10.1029/2006GB002751]. The enhancement of N(2) fixation and consequent C sequestration by tropical rivers appears to be a global phenomenon that is likely to be influenced by anthropogenic activity and climate change.

A novel method for the measurement of dissolved adenosine and guanosine triphosphate in aquatic habitats: applications to marine microbial ecology.

A novel method for the measurement of dissolved adenosine-5'-triphosphate and guanosine-5'-triphosphate (D-ATP and D-GTP, respectively) in marine and freshwater habitats was developed and applied to samples collected from the oligotrophic North Pacific Ocean. Both D-ATP and D-GTP are co-precipitated by authigenically formed Mg(OH)(2) and can be concentrated by factors greater than 200-fold, for subsequent measurement by the firefly luciferin-luciferase bioluminescence reaction. The detection limit for this method was 2-3 pmol ml(-1) of concentrated sample (equivalent to an in situ concentration of 10 pM) with a 5% precision at concentrations of 10 pmol ml(-1) or above.A significant positive correlation (P<0.001) was observed between particulate ATP (P-ATP) and D-ATP in water samples collected from Station ALOHA (22.75 degrees N, 158 degrees W; depth profiles 0-1000 m). The highest concentrations of dissolved nucleotides were found in the euphotic zone (0-175 m) below which the concentrations were low and relatively invariant. The dissolved nucleotide pools generally exceeded their corresponding particulate pools.Using radioisotopic tracer techniques and the new concentration method, turnover times for both particulate and dissolved nucleotides can be determined. The ability to measure concentrations and follow nucleotide tracers accurately in a very dilute environment provides a unique opportunity to address questions on microbial community metabolism, nutrient dynamics and energy flux.

Microorganisms in the accreted ice of Lake Vostok, Antarctica.

Analysis of a portion of Vostok ice core number 5G, which is thought to contain frozen water derived from Lake Vostok, Antarctica (a body of liquid water located beneath about 4 kilometers of glacial ice), revealed between 2 x 10(2) and 3 x 10(2) bacterial cells per milliliter and low concentrations of potential growth nutrients. Lipopolysaccharide (a Gram-negative bacterial cell biomarker) was also detected at concentrations consistent with the cell enumeration data, which suggests a predominance of Gram-negative bacteria. At least a portion of the microbial assemblage was viable, as determined by the respiration of carbon-14-labeled acetate and glucose substrates during incubations at 3 degrees C and 1 atmosphere. These accreted ice data suggest that Lake Vostok may contain viable microorganisms.

Prostaglandin synthetase inhibitors and dysmenorrhea. A survey and personal clinical experience.

A survey of earlier published studies on treatment of dysmenorrhea with prostaglandin synthetase inhibitors is given and personal clinical experiences are presented. The time when treatment should start in relation to the onset of bleeding is also discussed. A survey of studies published in English and Scandinavian literature yielded 532 patients. Pain relief was experienced in 64 to 100 per cent of the patients in these studies. The incidence of side-effects has generally been low but in a few studies a high incidence was reported. In the current study 34 patients with prinary dysmenorrhea completed the douule-blind, placebo controlled study on naproxen. The patients were treated for two cycles, 16 with naproxen and 18 with placebo. The mean relief score indicated a "slight to good" pain relief in the naproxen group and "no alleviation" in the placebo group. The difference is statistically significant (p = 0.003). Supplementary medication was much more used in the placebo group compared to the naproxen group (p = 0.01). In the placebo group no change whatsoever was demonstrated in alleviation of interference with every-day life, whereas there was a statistically significant improvement in the naproxen group. No major side-effect was registered. Thus none of the subjects withdrew from the study.