Comparing intra-articular CTXII levels assessed via magnetic capture or lavage in a rat knee osteoarthritis model.

OBJECTIVE: Parallel measures of osteoarthritis (OA) across species can help evaluate OA models relative to humans. Toward this need, our group recently developed a magnetic nanoparticle-based technology, termed magnetic capture, to analyze biomarkers within a rat knee. The objectives of this study were to directly compare magnetic capture to lavage, and assess c-telopeptide of collagen type II (CTXII) in the rat medial meniscus transection (MMT) model of knee OA. DESIGN: MMT surgery was performed in 30 male Lewis rats (3 months, 250 g). Using lavage or magnetic capture, CTXII was assessed in the OA-affected and contralateral knee at 1 week (n = 6 per group) or 4 weeks (n = 8 per group) after surgery. RESULTS: While lavage detected elevated CTXII concentrations in the OA-affected knee at 1 week (P = 0.002), magnetic capture detected elevated CTXII levels in the OA-affected knee at 4 weeks (P = 0.016). While magnetic capture did not detect significant elevation of CTXII at week 1, five of six rats evaluated with magnetic capture had higher CTXII levels in the OA-affected joint relative to the contralateral limb. Moreover, with magnetic capture, CTXII levels increased from 1 week to 4 weeks, corresponding to histological damage. CTXII concentrations evaluated via lavage were relatively constant across time. CONCLUSIONS: Magnetic capture and lavage evaluate CTXII in different ways: Magnetic capture measures total CTXII in the joint, while lavage measures concentration. Our data indicate magnetic capture may be advantageous at later time points, where CTXII can be diluted by effusions.

Unique spatiotemporal and dynamic gait compensations in the rat monoiodoacetate injection and medial meniscus transection models of knee osteoarthritis.

OBJECTIVE: In rodent osteoarthritis (OA) models, behavioral changes are often subtle and require highly sensitive methods to detect these changes. Gait analysis is one assay that may provide sensitive, quantitative measurement of these behavioral changes. To increase detection sensitivity of gait assessments relative to spatiotemporal gait collection alone, we combined our spatiotemporal and dynamic gait collection systems. Using this combined system, gait was assessed in the rat medial meniscus transection (MMT) model and monoiodoacetate (MIA) injection model of knee OA. DESIGN: 36 male Lewis rats were separated into MMT (n = 8), medial collateral ligament transection (MCLT) (n = 8), skin incision (n = 4), MIA injection (n = 8), and saline injection (n = 8) groups. After initiation of OA, gait data were collected weekly in each group out to 4 weeks. RESULTS: The MMT and MIA injection models produced unique pathologic gait profiles, with MMT animals developing a shuffling gait and MIA injection animals exhibiting antalgic gait. Spatiotemporal changes were also observed in the MMT model at week 1 (P < 0.01), but were not observed in the MIA injection model until week 3 (P < 0.01). Dynamic gait changes were observed in both models as early as 1 week post-surgery (P < 0.01). CONCLUSION: Combined analysis of spatiotemporal and dynamic gait data increased detection sensitivity for gait modification in two rat OA models. Analyzing the combined gait data provided a robust characterization of the pathologic gait produced by each model. Furthermore, this characterization revealed different patterns of gait compensations in two common rat models of knee OA.

OARSI Clinical Trials Recommendations: Design and conduct of implementation trials of interventions for osteoarthritis.

Rigorous implementation research is important for testing strategies to improve the delivery of effective osteoarthritis (OA) interventions. The objective of this manuscript is to describe principles of implementation research, including conceptual frameworks, study designs and methodology, with specific recommendations for randomized clinical trials of OA treatment and management. This manuscript includes a comprehensive review of prior research and recommendations for implementation trials. The review of literature included identification of seminal articles on implementation research methods, as well as examples of previous exemplar studies using these methods. In addition to a comprehensive summary of this literature, this manuscript provides key recommendations for OA implementation trials. This review concluded that to date there have been relatively few implementation trials of OA interventions, but this is an emerging area of research. Future OA clinical trials should routinely consider incorporation of implementation aims to enhance translation of findings.

Symptoms of the knee and hip in individuals with and without limb length inequality.

OBJECTIVE: This cross-sectional study examined the association of limb length inequality (LLI) with chronic joint symptoms at the hip and knee in a large, community-based sample, adjusting for the presence of radiographic osteoarthritis (OA) and other confounders. METHODS: The total study group comprised 3012 participants with complete knee symptoms data, 3007 participants with complete hip symptoms data, and 206 with LLI>or=2 cm. Presence of chronic knee symptoms was defined as report of pain, aching, or stiffness (symptoms) of the knee on most days. Presence of chronic hip symptoms was defined as hip pain, aching, or stiffness on most days or groin pain. Multiple logistic regression models were used to examine the relationship of LLI with knee and hip symptoms, while adjusting for demographic and clinical factors, radiographic knee or hip OA and history of knee or hip problems (joint injury, fracture, surgery, or congenital anomalies). RESULTS: Participants with LLI were more likely than those without LLI to have knee symptoms (56.8% vs 43.0%, P<0.001), and hip symptoms (49.5% vs 40.0%, P=0.09). In adjusted models, knee symptoms were significantly associated with presence of LLI (adjusted odds ratio [aOR]=1.41, 95% confidence interval, [95% CI] 1.02-1.97), but the relationship between hip symptoms and LLI (aOR=1.20, 95% CI 0.87-1.67) was not statistically significant. CONCLUSION: LLI was moderately associated with chronic knee symptoms and less strongly associated with hip symptoms. LLI may be a new modifiable risk factor for therapy of people with knee or hip symptoms.

Relationship of limb length inequality with radiographic knee and hip osteoarthritis.

OBJECTIVE: This study examined the relationship of limb length inequality (LLI) with radiographic hip and knee osteoarthritis (OA) in a large, community-based sample. METHODS: The total study group comprised 926 participants with radiographic knee OA, 796 with radiographic hip OA, and 210 (6.6%) with LLI >or=2cm. The presence of radiographic OA was defined as Kellgren/Lawrence (K/L) grade >or=2. Multiple logistic regression models were used to examine the relationship of LLI with hip and knee OA, while controlling for age, gender, race, body mass index, and history of hip or knee problems (joint injury, fracture, surgery, or congenital anomalies). RESULTS: In unadjusted analyses, participants with LLI were more likely than those without LLI to have radiographic knee OA (45.1% vs 28.3%, P<0.001) and radiographic hip OA (35.2% vs 28.7%, P=0.063). In multiple logistic regression models, knee OA was significantly associated with presence of LLI (adjusted Odds Ratio [aOR]=1.80, 95% Confidence Interval [95% CI] 1.29-2.52), but there was no significant relationship between hip OA and LLI (aOR=1.20, 95% CI 0.86-1.67). Among participants with LLI, right hip OA was more common when the contralateral limb was longer than when the ipsilateral limb was longer (30.3% vs 17.5%, P=0.070). CONCLUSION: LLI was associated with radiographic knee OA, controlling for other important variables. Future research should examine the relationship of LLI with hip or knee OA incidence, progression, and symptom severity, as well as the efficacy for LLI corrective treatments in OA.

Biochemical Characterization of Photosystem II Antenna Polypeptides in Grana and Stroma Membranes of Spinach.

The photosystem (PS) II antenna system comprises several biochemically and spectroscopically distinct complexes, including light-harvesting complex II (LHCII), chlorophyll-protein complex (CP) 29, CP26, and CP24. LHCII, the most abundant of these, is both structurally and functionally diverse. The photosynthetic apparatus is laterally segregated within the thylakoid membrane into PSI-rich and PSII-rich domains, and the distribution of antenna complexes between these domains has implications for antenna function. We report a detailed analysis of the differences in the polypeptide composition of LHCII, CP29, and CP26 complexes associated with grana and stroma thylakoid fractions from spinach (Spinacia oleracea L.), making use of a very high-resolution denaturing gel system, coupled with immunoblots using monospecific antibodies to identify specific antenna components. We first show that the polypeptide composition of the PSII antenna system is more complex than previously thought. We resolved at least five type I LHCII apoproteins and two to three type II LHCII apoproteins. We also resolved at least two apoproteins each for CP29 and CP26. In state 1-adapted grana and stroma thylakoid membranes, the spectrum of LHCII apoproteins is surprisingly similar. However, in addition to overall quantitative differences, we saw subtle but reproducible qualitative differences in the spectrum of LHCII apoproteins in grana and stroma membrane domains, including two forms of the major type II apoprotein. The implications of these findings for models of PSII antenna function in spinach are discussed.

Biogenesis of thylakoid membranes is controlled by light intensity in the conditional chlorophyll b-deficient CD3 mutant of wheat.

Biogenesis of thylakoid membranes in the conditional chlorophyll b-deficient CD3 mutant of wheat is dramatically altered by relatively small differences in the light intensity under which seedlings are grown. When the CD3 mutant is grown at 400 microE/m2 S (high light, about one-fifth full sunlight) plants are deficient in chlorophyll b (chlorophyll a/b ratio greater than 6.0) and lack or contain greatly reduced amounts of the chlorophyll a/b-binding complexes CPII/CPII (mobile or peripheral LHCII), CP29, CP24 and LHCI, as shown by mildly denaturing 'green gel' electrophoresis, by fully denaturing SDS-PAGE, and by Western blot analysis. High light CD3 chloroplasts display an unusual morphology characterized by large, sheet-like stromal thylakoids formed into parallel unstacked arrays and a limited number of small grana stacks displaced toward the edges of the arrays. Changes in the supramolecular organization of CD3 thylakoids, seen with freeze-fracture electron microscopy, include a reduction in the size of EFs particles, which correspond to photosystem II centers with variable amounts of attached LHCII, and a redistribution of EF particles from the stacked to the unstacked regions. When CD3 seedlings are grown at 150 microE/m2 S (low light) there is a substantial reversal of all of these effects. Thus, chlorophyll b and the chlorophyll a/b-binding proteins accumulate to near wild-type levels (chlorophyll a/b ratio = 3.5-4.5) and thylakoid morphology is more nearly wild type in appearance. Growth of the CD3 mutant in the presence of chloramphenicol stimulates the accumulation of chlorophyll b and its binding proteins (Duysen, M. E., T. P. Freeman, N. D. Williams, and L. L. Huckle. 1985. Plant Physiol. 78:531-536). We show that this partial rescue of the CD3 high light phenotype is accompanied by large changes in thylakoid structure. The CD3 mutant, which defines a new class of chlorophyll b-deficient phenotype, is discussed in the more general context of chlorophyll b deficiency.

Detection of type 1 insulinlike growth factor (IGF) receptors in Wilms' tumors.

Insulinlike growth factors (IGFs) are peptide hormones that regulate proliferation and differentiation of several types of normal and neoplastic cells. Recent studies of Wilms' tumor, a childhood kidney neoplasm, have detected increased expression of IGF-2 mRNA and protein. The present report describes detection of Type 1 IGF receptors in specimens of Wilms' tumor and adjacent nonneoplastic kidney tissue. These receptors recognize both IGF-1 and IGF-2, and binding of either peptide activates endogenous tyrosine kinase activity of the Type 1 IGF receptor beta subunit. These data indicate that Wilms' tumors contain receptors that recognize and respond to exogenous IGF in vitro, and that autocrine IGF production might contribute to the increased proliferation and abnormal differentiation of these cells in vivo.