Sandwich ELISA system for cartilage oligomeric matrix protein in equine synovial fluid and serum

Reasons for performing study: Measurement of cartilage oligomeric matrix protein (COMP) in serum has potential for diagnosis of equine osteoarthritis (OA), but clinical use is currently limited by the lack of specificity of an inhibition ELISA as well as by baseline increases due to exercise. Improved methods for ELISA with increased antigen specificity and sensitivity are therefore required for reliable measurement. Hypothesis: Measurement of the serum level of COMP by sandwich ELISA allows identification of horses with OA. Methods: New monoclonal antibodies (mAbs) were elicited against equine cartilage COMP, their epitopes were determined and a sandwich ELISA was developed. The concentrations of COMP in synovial fluid (SF; n = 100) and sera (n = 100) from OA cases were measured by sandwich ELISA as well as by inhibition ELISA and compared with concentrations in normal joints (n = 95) and horses (n = 50). Results: Immunoblots of enzymatically cleaved COMP showed that the new mAbs recognised different epitopes located on a 20 kDa fragment between K⁶³ and K²³⁸ of the EGF‐like repeats. Inhibition ELISA with any mAb detected significantly increased levels of COMP in OA SF compared with normal SF, whereas no significant difference was detected between serum levels of COMP in OA and normal horses. Conversely, sandwich ELISA with the combination of unlabelled 2A11 × biotinylated 11F10 mAbs detected a significant increase in COMP levels in both serum and SF from OA cases compared with levels in normal animals. Conclusions and potential relevance: Measurement of serum COMP with sandwich ELISA may be useful in identifying horses with OA.     

Efficacy of open patch-grafting under cardiopulmonary bypass for pulmonic stenosis in small dogs

Objective   To assess the efficacy of an open patch-graft technique under cardiopulmonary bypass (CPB) in small dogs.
Design and methods   A retrospective analysis of 10 dogs with pulmonic stenosis. Records between 1992 and 2002 were reviewed. The effect of surgical correction was evaluated and perioperative parameters were compared between survivors and non-survivors.
Results   The postoperative pulmonary pressure gradient was reduced in all seven surviving patients. Mean ± SE was 21.5 ± 7.4 mmHg (range 3.0–54.2 mmHg) and 6/7 dogs were < 40 mmHg at 3 months postoperatively. Comparing the data between those patients that survived and those that did not, the preoperative pressure gradient (P = 0.04) and volume of the Glucose-Insulin-Kalium solution used (P = 0.001) were significantly higher in those that did not survive.
Conclusion   Open patch-grafting can be performed in small-breed dogs and decreased the pulmonary pressure gradient in survivors at 3 months postoperatively. However, this technique is more invasive than balloon valvuloplasty and should be used cautiously in severely stenosed patients.     

Water adsorption process of bamboo heated at low temperature

Water adsorption capacities were evaluated for moso bamboo samples that were heated at 200°C for various times and conditioned in a closed container at 97% relative humidity at 20°C. Logistic regression analysis was used for curve fitting to the adsorption data and its parameters were analyzed. These parameters were compared with those derived previously from the Dubinin and Radushkevich theory. The properties of the heat-treated samples changed after 5 h of heating. With less than 5 h of heating, hydroxyl groups provided the main adsorption sites but their numbers decreased on heating. After 5 h, gasifi cation of the bamboo increased and capillaries formed. Part of this article was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, August 2007     

First application of PCR-Luminex system for molecular diagnosis of fungicide resistance and species identification of fungal pathogens

Fungicide resistance in plant pathogens is often caused by a single point mutation in a gene encoding fungicide target proteins. Such is the case for resistance to MBI-D (inhibitors of scytalone dehydratase in melanin biosynthesis) fungicides in rice blast fungus (Magnaporthe oryzae), which is caused by a mutation in the scytalone dehydratase gene that results in a replacement of valine with methionine at codon 75 of the fungicide target protein. PCR-Luminex, a novel system developed for high-throughput analysis of single nucleotide polymorphisms (SNPs) was successfully introduced to diagnose MBI-D resistance using specific oligonucleotide probes coupled with fluorescent beads. The PCR-Luminex system was further tested for its potential in identifying species causing Fusarium head blight on wheat. Four major pathogens, Fusarium graminearum (=F. asiaticum), F. culmorum, F. avenaceum, and Microdochium nivale, known to cause the disease, were tested, and the species were identified using the PCR-Luminex method. So far, this report is the first on the application of the DNA-based PCR-Luminex system in the area of crop protection and/or agricultural sciences.     

Relation between pathogenicity and genetic variation within <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis>

The relation between diversity of pathogenicity on clubroot-resistant (CR) cultivars of Chinese cabbage (Brassica rapa subsp. pekinensis) bred in Japan and DNA polymorphisms in 17 populations of Plasmodiophora brassicae from cruciferous plants was examined by inoculation tests and random amplified polymorphic DNA (RAPD) analysis using 18 arbitrary primers. Four pathotypes (A–D) were identified after inoculation of six CR cultivars of Chinese cabbage in the 17 populations from cruciferous crops. A relatively high level of genetic diversity was also detected among these populations in the RAPD analysis. Although the four pathotypes could not be clearly differentiated using the RAPD data, most populations of three pathotypes had a consistent location on the dendrogram. All pathotype B (virulent on five cultivars except Utage 70) and D (avirulent on all cultivars) populations, which were common in incompatible interactions with cv. Utage 70, were located in a single subcluster. All five pathotype C populations (virulent only on cv. Utage 70) except for one population grouped in another single subcluster. Because four pathotype A populations (virulent on all six cultivars, races 4 and 9) fell in different subclusters, the populations may be genetically polyphyletic. Populations from cruciferous weed Cardamine flexuosa differed remarkably from those from cruciferous crops in pathogenicity on common cultivars of Chinese cabbage and turnip and C. flexuosa, but they grouped in a single cluster with all race 9 populations from crops. Race 9 populations from crops may thus be closely related to populations from the weed rather than to races 1 and 4 from crops.     

Effects of tightening speed on torque coefficient in lag screw timber joints with steel side plates

To investigate the effects of tightening speed on the torque coefficient in lag screw timber joints with steel side plates, tightening tests were conducted on main timber members made from Cryptomeria japonica, Chamaecyparis obtusa and Pseudotsuga menziesii, under four tightening speed conditions (1, 4, 10, and 20 rpm). Major stick-slip behavior was observed in C. obtusa based on the relationship of tightening angle with clamp force, tightening torque, and thread torque at tightening speeds of 1 and 4 rpm. In addition, tightening speed’s effects on the torque coefficient (K) varied depending on the wood species of the main member. In P. menziesii, K was not affected by the tightening speed: the ratio of torque expended on tightening was 25% on average, and the ratio of torque expended on bearing surface friction was higher than the ratio of torque expended on thread friction.     

Direct observation and quantification of CO₂ binding within an amine-functionalized nanoporous solid

Understanding the molecular details of CO(2)-sorbent interactions is critical for the design of better carbon-capture systems. Here we report crystallographic resolution of CO(2) molecules and their binding domains in a metal-organic framework functionalized with amine groups. Accompanying computational studies that modeled the gas sorption isotherms, high heat of adsorption, and CO(2) lattice positions showed high agreement on all three fronts. The modeling apportioned specific binding interactions for each CO(2) molecule, including substantial cooperative binding effects among the guest molecules. The validation of the capacity of such simulations to accurately model molecular-scale binding bodes well for the theory-aided development of amine-based CO(2) sorbents. The analysis shows that the combination of appropriate pore size, strongly interacting amine functional groups, and the cooperative binding of CO(2) guest molecules is responsible for the low-pressure binding and large uptake of CO(2) in this sorbent material.     

The relationship between the size of caudolateral curvilinear osteophyte of the canine femoral neck and the radiographic view

Caudolateral curvilinear osteophyte (CCO), an osteophyte at the site of joint capsule attachment on the caudal aspect of the femoral neck, has been advocated as a radiographic criterion for coxofemoral subluxation. The correlation between the presence of CCO on radiographs (radiographic-CCO), the size of the CCO (CCO index) on three-dimensional computed tomographic (CT) images, and hip evaluation using transverse CT images was assessed in 22 Border Collies. CCOs were detected on the radiographs and CT images of 32% and 100% femurs, respectively. The CCO index correlated significantly with radiographic-CCO, but a large CCO index did not necessarily imply that the CCO was visible on radiographs. Hence, radiographic-CCO findings should be used cautiously in hip evaluation of Border Collies.     

End-of-day far-red treatment enhances responsiveness to gibberellins and promotes stem extension in chrysanthemum

SUMMARY

A brief end-of-day (EOD) far-red (FR) exposure promotes extension growth in plants. This change suggests a role for gibberellins (GA). For chrysanthemum (Chrysanthemum morifolium Ramat.), we show that EOD-FR exposure enhances shoot extension and is mediated, at least in part, by increased responsiveness (sensitivity) to GA. In addition, our analysis showed how light, especially EOD-FR, influenced shoot extension. The effect of EOD-FR on promoting extension growth was maintained, not only during the dark period, but also in the subsequent light period. Furthermore, our observations suggest that dark reversion of certain phytochromes is involved in the determination of flowering time. We also discuss the applicability of EOD-FR treatment to reduce the overall duration of cultivation, while maintaining a sufficient stem height during cut chrysanthemum production.