Purification and Immunoprotection Evaluation of AaHIV from Complex Venom Metalloproteinases of Deinagkistrodon acutus

The aim of this study was to investigate the immunoprotective effects of AaHIV in mice. After purification, a 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS‐PAGE) was performed. Bicinchoninic acid was used to determine the molecular weight and concentration of AaHIV. AaHIV, venom complex (VC), and phosphate buffered saline (PBS) were subsequently used to immunize the mice three times, and the blood was sampled 1 week after the third immunization to determine the serum immunoglobulin G (IgG) antibody titer. A skin‐bleeding inhibition assay and toxin‐eliminating assay were performed on the immunized mice. The purity and concentration of AaHIV were 86.6% and 1.20 mg/mL, respectively. The AaHIV group exhibited higher antibody titers than the VC group. The survival rate of the AaHIV group (7/10) was significantly higher than that of the PBS group (0/10) (P = 0.0031). The high titer of antibodies induced by AaHIV partially neutralized the bleeding activity of the Deinagkistrodon acutus venom complex.     

New methods for the assessment of mite numbers and results obtained for several textile objects

Summary The assessment of mite numbers in textile objects is important with regard to measures aimed at mite control. The standard method is to take a dust sample and to separate the mites from the dust by means of flotation. The results of mite assessment can be improved using methods based on the natural mobility of the mites; the so-called Mobility Test makes use of the intrinsic movement tendencies of the mites, whereas the Heat Escape Method uses the mobility of the mites in connection with the effects of heat. An example concernig two jackets shows the difference in results of mite assessment when using the conventional method (dust sample and flotation) on the one hand, and the new methods (Mobility Test and Heat Escape Method) on the other. It is shown that the conventional method can only supply relative mite numbers. The new methods also enable the user to monitor more efficiently the use of acaricide agents in mite control. This is demonstrated using the example of two beds and a carpet taken from a children's bedroom. The considerably higher numbers of mites suggest that the risk of the occurence of mite allergens and their penetration into the indoor air is even greater than previously assumed.     

利用各世代小区平均数的加权最小二乘法在玉米数量性状遗传分析上的应用

本文根据作者在参考文献[3]所提出的利用各世代小区平均数估计遗传参数的加权最小二乘法对玉米穗粒重的遗传特性进行了分析。结果表明:(1)在本次试验中,可用加性-显性-二基因互作模型来描述玉米穗粒重的遗传特性;其加性效应[d]、加性效应与显性效应的互作[j]在α=0.01水平上显著,显性效应[h]、显性效应与显性效应的互作[l]在α=0.10水平上显著,加性效应与加性效应的互作[i]不显著;[d]与[h]的差异在α=0.10水平上不显著,但([d] [h])与([i] [j] [l])在α=0.05水平上显著;(2)与利用6个世代平均数相比,利用8个世代小区平均数的加权最小二乘法既节约、又灵敏。     

温阳化瘀法对月经周期紊乱患者p53,p21,MDM2蛋白表达的影响

目的:通过对月经周期紊乱患者的细胞周期相关基因的表达水平进行分析,得出温阳化瘀法在此病上的治疗效果。方法:选取我院妇科收治的月经周期紊乱患者120例,参照随机原则共分为2组,其中西医治疗组59例,给予醋酸甲羟孕酮片和克罗米芬口服;实验组61例,在西医治疗基础上给予温阳化瘀中药治疗,每日1剂;另外随机选取同期体检健康的60名女性作为正常对照组,在治疗结束后,应用免疫组化方法对全部受试者进行p53、p21以及MDM2蛋白表达检测,同时应用统计学软件对相关结果进行分析。结果:1p53和p21蛋白的阳性表达主要在细胞核,MDM2蛋白阳性表达定位在细胞核和(或)胞质;2正常子宫内膜组织中p53蛋白阳性表达率为(36.67%),p21表达率为(33.33%),显著高于月经周期紊乱患者,中药治疗组的p53蛋白阳性表达率为(19.67%),p21达率为(18.03%),显著高于西医治疗组,P0.05,差异有统计学意义;3正常子宫内膜组织中MDM2蛋白阳性表达率为(1.67%),显著低于月经周期紊乱患者,中药治疗组的MDM2蛋白阳性表达率为(32.77%),显著低于西医治疗组(50.85%),P0.05,差异有统计学意义。结论:与健康妇女相比,月经周期紊乱患者的p53和p21蛋白阳性表达率显著降低,MDM2蛋白阳性率显著升高,应用温阳化瘀法能够显著改善月经周期紊乱患者细胞周期相关基因的表达水平。     

Pitfalls in histone propionylation during bottom‐up mass spectrometry analysis

Despite their important role in regulating gene expression, posttranslational histone modifications remain technically challenging to analyze. For identification by bottom‐up MS, propionylation is required prior to and following trypsin digestion. Hereby, more hydrophobic peptides are generated enabling RP HPLC separation. When histone dynamics are studied in a quantitative manner, specificity, and efficiency of this chemical derivatization are crucial. Therefore we examined eight different protocols, including two different propionylation reagents. This revealed amidation (up to 70%) and methylation (up to 9%) of carboxyl groups as a side reaction. Moreover, incomplete (up to 85%) as well as a specific propionylation (up to 63%) can occur, depending on the protocol. These results highlight the possible pitfalls and implications for data analysis when doing bottom‐up MS on histones.     

一种利用通用图像处理软件提高黑白生物特征图质量的方法

生物分类学工作者发表分类学文章常常需要制作和处理黑白生物特征图.本文利用通用图像处理软件Photoshop和Ilhustrator的不同功能特点,结合扫描仪、数位板等电脑外设,总结了一套专门适用生物分类特征草图覆墨的操作方法.方法包括草图输入、绘制墨线路径、覆墨、绘制阴影、排版输出等步骤.     

The Dry Aerosol Deposition Device (DADD): An instrument for depositing microbial aerosols onto surfaces

Concerns surrounding the contamination of infrastructure and equipment with biowarfare agents have led to the development of antimicrobial surfaces/coatings that are designed to “self-sterilize.” Surfaces will likely be contaminated via an aerosol exposure and thus antimicrobial efficacy measurements should also be performed using biological aerosols. Standard methods that use microbial agents suspended in aqueous buffers may provide misleading results that overestimate the performance of the surface. A settling chamber is the most common instrument for applying biological aerosols to surfaces. However, settling chambers have some drawbacks (e.g., slow loading times, large footprint, variable loading, etc.) that make them undesirable for many applications. We have developed a Dry Aerosol Deposition Device (DADD) that uses impaction rather than settling to load surfaces with biological aerosols. The use of impaction allows for rapid and highly reproducible loading of microorganisms onto surfaces. We have demonstrated that the DADD can deliver both Bacillus atrophaeus spores and Staphylococcus aureus vegetative cells to glass coupons at concentrations exceeding 1 × 10⁴ CFU/cm². The average coefficient of variation (CV) for sample-to-sample loading within an experiment was 13.6% for spores and 6.1% for S. aureus cells. The DADD is also a relatively simple and inexpensive device that can easily be contained within a 4-foot biological safety cabinet.     

Flow cytometry-based assay for the activity of NAD(P)H oxidoreductases of the outer mitochondrial membrane

NAD(P)H oxidoreductases of the outer mitochondrial membrane (OMM) are able to activate various xenobiotics and stimulate the production of reactive oxygen species and the opening of the mitochondrial permeability transition pore. However, the role of these systems in the cell damage by xenobiotics and chemotherapeutic drugs is poorly understood because the methods for the selective assessment of their activity have not been elaborated and specific inhibitors are unknown. Here we propose a method for the semiquantitative assessment of the activity of NAD(P)H oxidoreductases of the OMM in intact and permeabilized cells that is based on the flow cytometry detection of dimethylbiacridene, a fluorescent product of two-electron reduction of lucigenin. The method uses the structural feature of mitochondrial organization: the proximity of the sites of one-electron reduction of lucigenin to cation radical (NAD(P)H oxidoreductases of the OMM) to the sites of its subsequent oxidation (cytochrome c oxidase). The inhibition of cytochrome c oxidase by cyanide selectively activates the dimethylbiacridene formation by oxidoreductases of the OMM but not by other cellular oxidoreductases. The proposed protocol allows one to assess the lucigenin reductase (two-electron) activity of NAD(P)H oxidoreductases of the OMM and to compare it with the activity of other cellular systems that can be used for the analysis of the role of these systems in the cell damage by xenobiotics and antitumor drugs.     

Interlaboratory comparison study of serum total testoserone measurements performed by mass spectrometry methods

Background

Though mass spectrometry (MS) assays are increasingly used for routine clinical measurements of serum total testosterone (TT), information about the variability of results is limited. This study assessed the variability of TT measurement results from routine MS assays.

Methods

Twenty serum samples (12 females, 8 males) were analyzed on 2 days by seven high performance liquid chromatography (HPLC), and one gas chromatography (GC)-tandem mass spectrometry (HPLC-MS/MS, GC-MS/MS) assays. Two samples (male and female) were provided in five replicates to assess the within-run variability. Results were compared against those obtained at National Institute of Standards and Technology (NIST). The within- and between-laboratory variability was assessed for each sample. Comparisons to the NIST results were performed using bias plot and Deming regression analysis.

Results

The overall coefficient of variation of the results obtained with MS assays was <15%CV at >1.53 nmol/L and <34%CV at 0.3 nmol/L. The between-assay variability was the major contributor to the overall variability. The assay precision was the highest (<3%CV) with assays using liquid-liquid extraction for sample preparation or GC-MS/MS. The mean percent difference to the reference assay was 11%. The slopes of Deming regression analysis of the MS assays were between 0.903 and 1.138 (correlation coefficient: >0.996). TT concentrations for one assay were above the measurement range.

Conclusions

The variability of TT measurement results among MS assays is substantially smaller than that reported for immunoassays. The type of sample preparation may affect assay precision. Standardizing assays can further reduce the variability of measurement results.