Validation,use, and disadvantages of enzyme-linked immunosorbent assay kits for detection of cortisol in channel catfish,largemouth bass,red pacu,and golden shiners

Measurement of cortisol response is an important tool to asses stress in fisheries research. Radioimmunoassay (RIA) is a common method for the measure of cortisol in fish. Use of enzyme-linked immunosorbent assays (ELISA) to detect cortisol would eliminate health hazards, costs of handling radioisotopes, and the short stability time associated with RIA. Enzyme-linked immunosorbent assays have been used for the determination of cortisol in several fish species. However, the ELISA method of cortisol determination in fish lacks proper validation testing. We conducted validation procedures for multiple commercial cortisol ELISA kits and compared the results to RIA. The assays were tested for four species: (1) channel catfish Ictalurus punctatus, (2) largemouth bass Micropterus salmoides, (3) red pacu Piaractus brachypomus, and (4) golden shiners Notemigonus crysoleucas. We evaluated the ELISA methods against RIA, and determined that at least one kit is suitable (accuracy: mean recovery of spiked samples, 102.8%; reproducibility: interassay coefficient of variation < 10.5% for all species; precision: intra-assay coefficient of variation < 16.8% for all species; linearity: R ² > 0.96 for all species) for the measurement of cortisol response in fish and comparative determination of stress. All of the ELISA assay results varied by more than 10% from the cortisol concentrations detected by the RIA. The high variability of the kit results indicates that commercial ELISA kits could be utilized for qualitative determination of cortisol in fish, but should be fully validated in each laboratory for each species before being used for research.     

刺参(Apostichopus japonicus)重要经济性状相关SNP标记的验证分析

本研究在前期构建的刺参(Apostichopus japonicus)高密度遗传连锁图谱和QTL分析的基础上,筛选出26个与体长、体重、体宽、棘刺总数、抗病力相关的SNP位点,设计出可用于HRM检测的SNP扩增引物13对。在扩大群体中利用HRM小片段法对这13个刺参重要经济性状相关的候选SNP位点进行分型和多态性检测,并结合扩大群体的相关性状数据进行了QTL位点验证。多态性结果显示,13个位点中有3个单态性位点,其余10个多态性位点中有3个位点为低等位多态性,7个位点为中等位多态性。10个多态性位点的最小等位基因频率(MAF)介于0.016(SNP113)~ 0.332(SNP160)之间,平均值为0.173;各位点的观测杂合度(Ho)介于0.031(SNP113)~0.818(SNP9)之间,平均值为0.433;期望杂合度(He)介于0.031(SNP113)~0.834(SNP9)之间,平均值为0.402;多态信息含量(PIC)介于0.030(SNP113)~0.393(SNP160)之间,平均为0.248,有6个位点偏离Hardy- Weinberg平衡。QTL验证结果表明,SNP40和SNP160位点为与生长(体长、体重、体宽)相关的位点,各位点的优势基因型分别为SNP40(CC)和SNP160(AA);SNP88、SNP112和SNP126这3个位点为与抗病力相关的位点,各位点的优势基因型为SNP88(CC)、SNP112(AA)和SNP126(TT)。基于这5个位点构建生长和抗病二倍型,发现二倍型K1(CC AA TT)抗病力最强,S1(CC AA)、S3(CC AC)在生长方面优势显著,相关研究结果可为分子标记辅助育种在生产中应用提供基础数据。     

车用磁流变液制动器制动效果分析与研究

以磁流变液(MRF)的流变特性为基础,推导了双盘式圆盘型磁流变液制动器的制动力矩计算公式;在整车环境下对磁流变液制动器进行了匹配设计,并在Matlab/Simulink软件环境下建立ABS制动控制系统模型;在ABS模型中,对磁流变液制动器的性能进行了仿真和分析,其结果证实磁流变液制动器具有优越的综合制动效果。     

Estimating chlorophyll content from hyperspectral vegetation indices: Modeling and validation

Leaf chlorophyll content, a good indicator of photosynthesis activity, mutations, stress and nutritional state, is of special significance to precision agriculture. Recent studies have demonstrated the feasibility of retrieval of chlorophyll content from hyperspectral vegetation indices composed by the reflectance of specific bands. In this paper, a set of vegetation indices belonged to three classes (normalized difference vegetation index (NDVI), modified simple ratio (MSR) index and the modified chlorophyll absorption ratio index (MCARI, TCARI) and the integrated forms (MCARI/OSAVI and TCARI/OSAVI)) were tested using the PROSPECT and SAIL models to explore their potentials in chlorophyll content estimation. Different bands combinations were also used to derive the modified vegetation indices. In the sensitivity study, four new formed indices (MSR[705,750], MCARI[705,750], TCARI/OSAVI[705,750] and MCARI/OSAVI[705,750]) were proved to have better linearity with chlorophyll content and resistant to leaf area index (LAI) variations by taking into account the effect of quick saturation at 670 nm with relatively low chlorophyll content. Validation study was also conducted at canopy scale using the ground truth data in the growth duration of winter wheat (chlorophyll content and reflectance data). The results showed that the integrated indices TCARI/OSAVI[705,750] and MCARI/OSAVI[705,750] are most appropriate for chlorophyll estimation with high correlation coefficients R² of 0.8808 and 0.9406, respectively, because more disturbances such as shadow, soil reflectance and nonphotosynthetic materials are taken into account. The high correlation between the vegetation indices obtained in the developmental stages of wheat and Hyperion data (R² of 0.6798 and 0.7618 for TCARI/OSAVI[705,750] and MCARI/OSAVI[705,750], respectively) indicated that these two integrated index can be used in practice to estimate the chlorophylls of different types of corns.     

产表面活性剂菌的筛选和验证实验研究

[目的]探究废弃钻井液中产表面活性剂菌株的性能.[方法]以废弃的钻井液为原材料,利用生物表面活性剂的性质进行产表面活性剂菌株的培养和筛选,并且将筛选出来的产表面活性剂菌进行排油活性和乳化性能的测定.[结果]筛选出的产表面活性剂菌排油圈的直径约为4 cm,乳化层初始较厚,1d后变薄.[结论]筛选出的产表面活性剂菌排油活性较好,但乳化性能不稳定.     

Permeability determination and pharmacokinetic study of nobiletin in rat plasma and brain by validated high-performance liquid chromatography method

In the present study, we are reporting permeability and pharmacokinetics of nobiletin in rat plasma and brain, using a validated reverse phase high performance liquid chromatographic method. Protein precipitation method was used for the extraction of nobiletin and coumarin (IS) from rat plasma and brain tissue. The system was run in isocratic mode with mobile phase consisting of potassium dihydrogen ortho-phosphate (pH 4.5; 0.04 mM) and acetonitrile in ratio of 50:50, v/v. The total chromatographic run time was 9.0 min. The method was proved to be accurate and precise at linearity range of 0.05–10 μg/mL with a correlation coefficient (r) of ≥ 0.994 in rat plasma and ≥ 0.995 in rat brain. The intra- and inter-day precision and accuracy values are found to be within the assay variability limits as per the FDA guidelines. Nobiletin was found stable in the battery of stability studies viz., bench-top, auto-sampler, freeze/thaw cycles and long term storage in a freezer at − 70 ± 10 °C. Maximum concentrations of nobiletin in both plasma and brain were observed at 1 h after single oral dosing (50 mg/kg). The maximum concentration in plasma and brain were 1.78 and 4.20 μg/mL, respectively. The AUC_0–t in plasma and brain were 7.49 and 20.66 μg·h/mL, respectively. The mean elimination half life (t_1/2) in plasma and brain were 1.80 and 11.42 h, respectively. The Parallel Artificial Membrane Permeability Assay (PAMPA) permeability of nobiletin was found to be high at both pH 4.0 and 7.0.     

Copper in the terrestrial environment: Verification of a laboratory-derived terrestrial biotic ligand model to predict earthworm mortality with toxicity observed in field soils

This study was set up for validation of a regression model to predict mortality in the terrestrial earthworm Aporrectodea caliginosa following exposure to copper. This model was derived from a terrestrial biotic ligand model and incorporates the protective effects of H⁺ and Na⁺ on copper toxicity.Three soil sets were used for the experiments, all of which had a different copper contamination history over more than 20 years and were considered to be aged field soils. The soils were characterized by analysis of various copper pools in the solid phase and in the pore water, analysis of physical and chemical soil properties and by regression analysis. Measured and calculated copper activities (expressed as pCu) correlated reasonably well. Measured copper activities correlated with the total copper concentration in the pore water and the pH. The organic matter in the solid phase had no influence on the pCu in these soil sets.Earthworms were exposed to the soils for 28 days, after which survival was scored. Observed earthworm mortality after 28-days exposure was plotted as a function of the log-transformed difference between predicted (log₁₀ transformed) LC50-values and measured values of pCu for validation of the regression model. The results obtained were in agreement with the assumption that mortality is to be observed in those soils where the predicted LC50 exceeds the measured pCu. However, a structural underestimation of toxicity was apparent, which is most likely due to mixture effects related to the presence of additional substances in field soils. Nevertheless, the trend of the results in the validation tests demonstrates that the newly developed toxicity model is a useful tool in predicting lethality of copper contamination to earthworms in field soils.     

塑料大棚小气候的模拟和检测

为了研究大棚内外空气热量的物质交换的机理，该文把用在大玻璃温室的让布鲁温室动态模型应用于小塑料大棚。此模型考虑了温室内三层土壤、作物层、空气层、覆盖物和保温幕之间的传导、对流、太阳辐射和热辐射以及潜热交换，在此基础上进行了在不同气候条件下模拟的实测结果的比较分析。其结果表明，模型达到较高精度。     

Development of a composite orthopaedic pain scale in horses

This study addresses development and validation of a composite multifactorial pain scale (CPS) in an experimental equine model of acute orthopaedic pain. Eighteen horses were allocated to control (sedation with/without epidural analgesia - mixture of morphine, ropivacaine, detomidine and ketamine) and experimental groups: amphotericin-B injection in the tarsocrural joint induced pain and analgesia was either i.v. phenylbutazone administered post-induction of synovitis, or pre-emptive epidural mixture, or a pre-emptive combination of the 2. Inter- and intra-observer reproducibility was good (0.8相似文献