Analytical and Clinical Validation of a Time-resolved Immunofluorometric Assay (TR-IFMA) for Canine C-reactive Protein in Serum

A time-resolved immunofluorometric assay (TR-IFMA) was developed for the determination of C-reactive protein (CRP) in canine serum. CRP was isolated from canine acute-phase serum by affinity chromatography on agarose coupled with phosphorylethanolamine. This isolated dog CRP was used as standard to calibrate the assay. Intra-assay and inter-assay coefficients of variation were in the ranges 5.3–7.1% and 4.8–13.3%, respectively. Accuracy, evaluated by adding 2 and 10 μg/ml of CRP to serum samples, provided recoveries of 99.9% and 106.8%. High correlation was found between CRP measurements by TR-IFMA and a by commercial enzyme-linked immunosorbent assay (R² = 0.98). The limit of detection for the TR-IFMA method was 0.000067 μg/ml and the measurement of CRP in serial dilutions of acute-phase dog sera generated curves with the same slope as the one constructed with purified CRP. The TR-IFMA provides a precise, accurate and highly sensitive assay for CRP determination in dog samples. CRP levels in dogs with different diseases ranged between 10.2 and 210.7 μg/ml and were significantly higher than those observed in healthy dogs (< 7.1 μmg/ml).     

Optimization and validation of a flow cytometric method for immunophenotyping peripheral blood lymphocytes from cynomolgus monkeys (Macaca fascicularis)

BACKGROUND: Guidelines published by the Food and Drug Administration and Center for Human Medicinal Products describe the need to assess immunotoxic effects in nonclinical studies that evaluate drug toxicity, including the use of immunophenotyping to measure immunotoxicity. We are not aware of previous studies, however, that have validated methods for immunophenotyping peripheral blood lymphocyte subsets in whole blood samples from cynomolgus monkeys. OBJECTIVE: The purpose of this study was to optimize and validate a flow cytometric assay for immunophenotyping lymphocytes in the peripheral blood of cynomolgus monkeys. METHODS: A series of prevalidation experiments were done to determine optimal reagents, volumes, timing, and other procedural details of the flow cytometric assay. Using the optimized method, we then determined precision, interindividual variation, laboratory-to-laboratory variability, and sample stability. Stabilized human blood was used as a positive control for staining, processing, and analysis. The percentage and number of pan-T cells (CD3+), T-helper cells (CD3+4+), T cytotoxic/suppressor cells (CD3+8+), natural killer cells (CD3-16+), and B-cells (CD3-20+) were determined in 146 male and 140 female, clinically healthy monkeys and reference intervals were calculated. RESULTS: By doing 4-color staining with a lyse-wash method, intra- and interassay precision were <5% for all lymphocyte subsets. Variability between technicians and laboratories was minimal (CVs<3%). Samples were stable for up to 24 hours after staining and fixing. CONCLUSIONS: The validated method is extremely robust and can be performed under good laboratory practice conditions to support nonclinical studies. Reference intervals for lymphocyte subsets were similar to those previously reported.     

Validation of an immunoturbidimetric D-dimer assay in canine citrated plasma

Abstract: D-dimer is a neoantigen formed when thrombin initiates the transformation of fibrinogen to fibrin; it is derived from plasmin digestion of cross-linked fibrin. In human medicine, the usefulness of this analyte in diagnosing disseminated intravascular coagulation (DIC) has been assessed in patients fulfilling the clinical and laboratory requirements for this disorder. In canine medicine, the use of D-dimer is relatively new. Detailed studies are needed to understand the relationship between D-dimer concentration in plasma and DIC status in dogs. We validated a D-dimer immunoturbidimetric assay (Tina-quant [a] D-Dimer, Boehringer Mannheim) in canine citrated plasma samples. Intra-assay and interassay variability (coefficient of variation) was 5.63% and 8.82%, respectively. The assay was linear, using 2 samples with low and high D-dimer concentrations (r = .996 and .998). Accuracy was 102.2% and 95.7% based on a recovery study in which 2 samples were assessed. Reference values for D-dimer were established using 70 healthy dogs that were assessed clinically and evaluated on the basis of a complete laboratory workup. The reference range was set between 0.02 and 0.28 μg/mL (chi-square test for normal distribution, P > .05).     

应用形态度量学方法对中华纹胸鮡和福建纹胸鮡物种有效性的研究

采用形态度量学方法对分布于长江、珠江、九龙江、海南岛和闽江的福建纹胸和中华纹胸 12 6个个体框架结构测量中的 2 3个指标进行了主成分分析 ,以探求中华纹胸和福建纹胸的物种有效性。初步研究结果表明各样本贡献值较大的前 4个主成分在数值上并没有明显的差异。显然 ,以上 2个物种在形态上并没有明显的区别 ,福建纹胸应为中华纹胸的同物异名 ,而不应为一个独立的物种     

Evaluation of the agreement of two oscillometric blood pressure devices with invasive blood pressure in anaesthetized chimpanzees (Pan troglodytes)

ObjectiveTo evaluate the agreement of two noninvasive blood pressure devices: a human device with the cuff placed on the wrist (Omron R1) and a veterinary device with the cuff placed on the upper brachium (Surgivet Advisor Vital Signs Monitor) with invasive blood pressure (IBP) measurement in anaesthetized chimpanzees.Study designProspective clinical study.AnimalsA convenience sample of 11 adult chimpanzees undergoing anaesthesia for translocation and routine health checks.MethodsSystolic (SAP) and diastolic arterial pressures (DAP) were continuously recorded via a transducer connected to a femoral artery cannula, and at 5 minute intervals from the two oscillometric devices. Agreement was explored using Bland-Altman analysis and bias defined as the mean difference between the two measurement methods. Spearman correlation coefficients were calculated. Significance was set at p < 0.05.ResultsBias and standard deviation for the Surgivet compared with IBP were 8.6 ± 18 for SAP and 8.4 ± 9.9 for DAP, showing a significant underestimation of both variables. Limits of agreement (LOA) were from –27 to 44 for SAP and from –11 to 28 for DAP. Correlation coefficients between the Surgivet and IBP values were 0.86 for SAP and 0.85 for DAP (p < 0.0001). Bias and standard deviation for the Omron compared with the IBP were –21 ± 25 for SAP and –18 ± 15 for DAP, showing a significant overestimation of both variables. LOA were from –70 to –28 for SAP and from –47 to 11 for DAP. Spearman correlation coefficients between the Omron and IBP values were 0.64 for SAP and 0.72 for DAP (p < 0.0001).Conclusions and clinical relevanceAlthough neither device met all the criteria for device validation, the Surgivet presented better agreement with IBP values than the Omron in adult anaesthetized chimpanzees.     

Across-breed validation study confirms and identifies new loci associated with sexual precocity in Brahman and Nellore cattle

The aim of this study was to identify candidate regions associated with sexual precocity in Bos indicus. Nellore and Brahman were set as validation and discovery populations, respectively. SNP selected in Brahman to validate in Nellore were from gene regions affecting reproductive traits (G1) and significant SNP (p ≤ 10^–3) from a meta-analysis (G2). In the validation population, early pregnancy (EP) and scrotal circumference (SC) were evaluated. To perform GWAS in validation population, we used regression and Bayes C. SNP with p ≤ 10^–3 in regression and Bayes factor ≥3 in Bayes C were deemed significant. Significant SNP (for EP or SC) or SNP in their ±250 Kb vicinity region, which were in at least one discovery set (G1 or G2), were considered validated. SNP identified in both G1 and G2 were considered candidate. For EP, 145 SNP were validated in G1 and 41 in G2, and for SC, these numbers were 14 and 2. For EP, 21 candidate SNP were detected (G1 and G2). For SC, no candidate SNP were identified. Validated SNP and their vicinity region were located close to quantitative trait loci or genes related to reproductive traits and were enriched in gene ontology terms related to reproductive success. These are therefore strong candidate regions for sexual precocity in Nellore and Brahman.     

基于优化支持向量机的采煤机故障诊断技术

为了对采煤机故障进行准确诊断研究,本文提出了一种基于优化支持向量机的采煤机故障诊断新方法,首先采用主成分分析法(PCA)对采煤机故障特征参数进行特征提取,其次应用特征数据进行基于支持向量机(SVM)的采煤机故障诊断模型训练,再次采用交叉验证方法对SVM模型参数进行优化,建立最优SVM采煤机故障诊断模型,最后通过对比实验,验证了基于优化SVM采煤机故障诊断模型的可行性和优越性。     

基于响应面法的棉秆压捆试验研究

针对棉秆压缩打捆密度低、压缩打捆装备可靠性差等问题,采用设计的棉秆压捆试验平台,将棉秆含水率、棉秆切断长度、棉秆喂入量和压缩活塞压缩频率作为试验因素,以压缩活塞端面压力、压缩室压力和棉秆压捆密度作为试验性能指标进行棉秆压缩打捆单因素和中心组合试验,建立各试验因素和试验性能指标之间的回归方程,确定各试验因素对性能指标的影响规律,并进行优化计算,对优化结果进行试验验证。结果表明:棉秆压缩打捆的最优组合为棉秆含水率30%,棉秆切断长度25 cm,棉秆喂入量2.15 kg/s,压缩活塞压缩频率110次/min,压缩活塞端面压力13.84 kN,压缩室压力3.36 kN,棉秆打捆密度145.83 kg/m~3,为棉秆压缩打捆机械的设计提供理论指导。     

Life-history characteristics of an age-validated established invasive African sharptooth catfish,Clarias gariepinus,population in a warm–temperate African impoundment

It has been suggested that the invasive, omnivorous Clarias garipienus is capable of rapid invasions and long-term persistence in recently inhabited freshwater systems. To test this hypothesis, the life history of the established, extralimital Darlington Dam (33°10’31”S, 25°09’13”E) population was investigated. By counting post-fluorescent mark increments on otoliths from 21 chemically tagged wild fish recaptured 244–537 days later, the deposition of growth zones, comprising alternating opaque and translucent bands, was validated as annual. Examination of sectioned otoliths from 175 fish revealed that the oldest fish, two males of 840 and 1074 mm total length (TL), were 25 years old – 10 years older than previously described for any C. gariepinus population. The oldest female was 885 mm TL and 21 years old. Length-at-age was subsequently described using the von Bertalanffy growth model. Combined-sex growth was best described as L_t = 931.7(1– exp(–0.15(t+2.43))) mm TL. Total mortality (Z) was calculated using catch curve analysis and the Chapman & Robson estimator to be 0.35/yr. The presence of specimens 15 years and older indicates that these fish established quicklyand supports the finding that mortality rates are low, which, in turn, suggests likely long-term population persistence.