Equine synovial fluid small non-coding RNA signatures in early osteoarthritis.

BACKGROUND: Osteoarthritis remains one of the greatest causes of morbidity and mortality in the equine population. The inability to detect pre-clinical changes in osteoarthritis has been a significant impediment to the development of effective therapies against this disease. Synovial fluid represents a potential source of disease-specific small non-coding RNAs (sncRNAs) that could aid in the understanding of the pathogenesis of osteoarthritis. We hypothesised that early stages of osteoarthritis would alter the expression of sncRNAs, facilitating the understanding of the underlying pathogenesis and potentially provide early biomarkers. METHODS: Small RNA sequencing was performed using synovial fluid from the metacarpophalangeal joints of both control and early osteoarthritic horses. A group of differentially expressed sncRNAs was selected for further validation through qRT-PCR using an independent cohort of synovial fluid samples from control and early osteoarthritic horses. Bioinformatic analysis was performed in order to identify putative targets of the differentially expressed microRNAs and to explore potential associations with specific biological processes. RESULTS: Results revealed 22 differentially expressed sncRNAs including 13 microRNAs; miR-10a, miR-223, let7a, miR-99a, miR-23b, miR-378, miR-143 (and six novel microRNAs), four small nuclear RNAs; U2, U5, U11, U12, three small nucleolar RNAs; U13, snoR38, snord96, and one small cajal body-specific RNA; scarna3. Five sncRNAs were validated; miR-223 was significantly reduced in early osteoarthritis and miR-23b, let-7a-2, snord96A and snord13 were significantly upregulated. Significant cellular actions deduced by the differentially expressed microRNAs included apoptosis (P < 0.0003), necrosis (P < 0.0009), autophagy (P < 0.0007) and inflammation (P < 0.00001). A conservatively filtered list of 57 messenger RNA targets was obtained; the top biological processes associated were regulation of cell population proliferation (P < 0.000001), cellular response to chemical stimulus (P < 0.000001) and cell surface receptor signalling pathway (P < 0.000001). CONCLUSIONS: Synovial fluid sncRNAs may be used as molecular biomarkers for early disease in equine osteoarthritic joints. The biological processes they regulate may play an important role in understanding early osteoarthritis pathogenesis. Characterising these dynamic molecular changes could provide novel insights on the process and mechanism of early osteoarthritis development and is critical for the development of new therapeutic approaches.

Reliability of the Tardieu Scale for assessing spasticity in children with cerebral palsy.

OBJECTIVE: To measure the Tardieu Scale's reliability in children with cerebral palsy (CP) when used by raters with and without experience in using the scale, before and after training. DESIGN: Single-center, intrarater and interrater reliability study. SETTING: Institutional ambulatory care. PARTICIPANTS: Referred children with CP in the pretraining phase (n=5), during training (n=3), and in the posttraining phase (n=15). INTERVENTIONS: The Tardieu Scale involves performing passive muscle stretch at 2 velocities, slow and fast. The rater derives 2 parameters; the Spasticity Angle X is the difference between the angles of arrest at slow speed and of catch-and-release or clonus at fast speed; the Spasticity Grade Y is an ordinal variable that grades the intensity (gain) of the muscle reaction to fast stretch. In phase 1, experienced raters without formalized training in the scale graded elbow, knee, and ankle plantar flexors bilaterally, without and with a goniometer. In phase 2, after training, the experienced and nonexperienced raters graded the same muscles unilaterally. MAIN OUTCOME MEASURES: Intrarater and interrater reliability of the Tardieu Scale. RESULTS: After training, nonexperienced raters had mean +/- SD intrarater and interrater agreement rates across all joints and parameters of 80%+/-14% and 74%+/-16%, respectively. For experienced raters, intrarater and interrater agreement rates before training were 77%+/-13% and 66%+/-15%, respectively, versus 90%+/-8% and 81%+/-13%, respectively, after training (P<.001 for both). Specific angle measurements at the knee were less reliable for the angles of catch measured at fast speed. Across all joints, agreement rates were similar using visual or goniometric measurements. CONCLUSIONS: Both parameters of the Tardieu Scale have excellent intrarater and interrater reliability when assessed at the elbow and ankle joints of children with CP, with no difference noted between visual and goniometric measurements. Angle measurements were less reliable at the knee joints. Training was associated with a highly significant improvement in reliability.

Patient attitudes toward screening.

BACKGROUND: Physician organizations recommend screening for health care behaviors. Despite these recommendations, health care providers worry that questions on sensitive topics may not be accepted by their patients. To determine if there is a relationship between health care screening by providers and acceptance of that screening by patients, a survey of female patients was analyzed. METHOD: Two telephone surveys were conducted two years apart. Each was a cross-sectional sample of female patients over the age of 18 years who had been seen by their primary care provider (PCP) in the previous 12 months. Patients were asked if they had been screened for eight different health behaviors (exercise, smoking, use of alcohol or drugs, excessive stress, sexual functioning concerns, safety or violence in the home, guns in the home) in the past year. They were also asked about their attitudes toward screening for those behaviors by health care providers. Odds ratios were calculated for patients who both agreed that screening should occur and reported having been screened in the last year. RESULTS: 3,175 women were surveyed. There was high acceptance of routine screening for exercise (75%), smoking (72%), alcohol/drugs (68%), and stress (62%), but less for sexual functioning (40%), safety/violence (40%), or guns (23%). There was a higher likelihood of agreeing with routine screening if the patient reported having been screened in the past year: exercise (OR 2.3, 95% CI 1.8-2.9), smoking (OR 1.6, 95% CI 1.3-1.9), alcohol/drugs (OR 2.3, 95% CI 1.9-2.7), stress (OR 1.7, 95% CI 1.4-1.9), sexual functioning (OR 2.7, 95% CI 2.2-3.4), safety/violence (OR 3.4, 95% CI 2.8-4.2), and guns (OR 4.4, 95% CI 3.4-5.8). LIMITATIONS: Only women in established relationships with primary care providers were surveyed. The cross-sectional nature of the survey prevents determination of the causality of the relationship. CONCLUSION: Women who had been screened for a health behavior had greater acceptance of routine screening for that behavior. Although further research is needed to determine the casual relationship, providers should not worry about offending their patients when screening for sensitive health behaviors.