金针菇中蛋白质含量的变化和其中一个蛋白质的生物活性

在对金针菇(Flammulinavelutipes)子实体活性蛋白质纯化过程中发现,金针菇在不同环境条件下不同蛋白质的表达量有差异.通过阴离子交换层析、凝胶层析和高压液相色谱,从金针菇中分离纯化获得了一种蛋白质Zb.经SDS-PAGE测定该蛋白分子量(Mr)为 30×103,等电聚焦电泳表明该蛋白质的等电点为 4. 7,且该蛋白质不含糖.N-端测序表明,该蛋白质N-端的 20个氨基酸为PQVKTSWEDLANLGWPIQQV.此外,在HepG2. 2. 2. 15细胞株上检测该蛋白质体外对肝炎病毒的抑制作用,发现对HBsAg抑制作用明显,抑制中浓度为 0. 117μg/mL,但对HBeAg几乎无作用.以胃癌细胞株MGC80-3为研究对象,检测表明,该蛋白质对MGC80-3抑制率在 50%时的蛋白质浓度约为 0. 75μg/mL.以起始浓度为 0. 296mg/mL的Zb检测它的血凝活性时发现,Zb对兔血红细胞和人A、B、O血红细胞的血凝滴度为 2-12.但同时观察到Zb处理后的兔血红细胞形态发生改变. 图 6表 1参 15     

新型多级冲击采样器切割粒径的标准粒子实验分析

大气溶胶对环境污染、气候变化以及人体健康有着重要的影响,大气溶胶的采集和分析已成为当前大气环境研究领域中的一个重要课题.论述的新型多级冲击采样器能够实现大气溶胶颗粒物按空气动力学粒径的大小7级分离采集,提供大致均匀的平面样品以进行后续的物理、化学分析,同时保证各分级切割粒径的稳定性.利用振动孔溶胶发生器产生均匀、粒径大致相同、单分散相的1.5～17μm空气动力学粒径标准粒子,对新型多级冲击采样器前三级的切割粒径和收集效率进行鉴定性分析实验,结果表明,能够满足对大气溶胶颗粒物分级采集的要求.     

羟基致死船舶压载水微生物的生化影响研究

试验表明羟基比值浓度达到0．63mg／L左右时，足以杀死压载水微生物。在此浓度下也会引起微生物的糖、脂质、蛋白质、核酸、酶等发生强烈的化学反应。脂质过氧化物含量上升为原来的3倍；总蛋白质减少了33．4％；核酸中RNA、DNA分别减少了46．9％、77．0％；葡萄糖却增加了1倍，羟基破坏了多糖，使之降解为单糖；过氧化物酶含量减少了21．9％；超氧化物歧化酶减少了54．2％；分离微生物后海水中过氧化氢酶、超氧化物歧化酶分别减少了83．1％、33．2％，上述生化试验资料清楚表明了羟基对微生物三大基本分子(核酸、蛋白质、葡萄糖)、抗氧化酶系(过氧化酶、过氧化氢酶、超氧化物歧化酶)以及脂质均有强烈的破坏作用，这也是致死压载水微生物的主要原因。     

久效磷对扁藻的生理效应

运用急性毒性实验和生化分析方法对久效磷胁迫下扁藻细胞的生理变化进行了研究，结果表明：（１）久效磷对扁藻生长有较强的抑制作用。其７２ｈ半抑制剂量为１．４６ｍｇ／Ｌ；（２）久效磷胁迫下，扁藻细胞的３种大分子物质－－蛋白质、ＤＮＡ和ＲＮＡ的含量降低；（２）久效磷引起扁藻叶绿素ａ的降解和光合速率的下降，这说明久效磷处理抑制了扁藻的生理代谢。     

稀土金属离子对梨形四膜虫的24小时生长影响

应用４种方法（细胞直计计数法、中性红试验、重蛋白及核酸含量测定）研究不同浓度稀土金属离子Ｌａ＾２＋、Ｓｍ＾３＋、Ｙ＾３＋、Ｇｄ＾３＋处理２４ｈ后，对梨形四膜虫的生长影响，结果表明，在低浓度时Ｌａ＾３＋、Ｓｍ＾３＋、Ｙ＾３＋，对四膜虫具有刺激生长的作用。随着浓度的升高，４种稀土金属离子均表现出毒性效应；高浓度时，抑制生长艇特别明显。计算Ｌａ＾３＋、Ｓｍ＾３＋、Ｙ＾３＋、Ｇｄ＾３＋的半数抑制浓度（ＩＣ     

Lipophilic organic pollutants induce changes in phospholipid and membrane protein composition leading to Vero cell morphological change

Membrane damage related to morphological change in Vero cells is a sensitive index of the composite biotoxicity of trace lipophilic chemicals. However, judging whether the morphological change in Vero cells happens and its ratio are difficult because it is not a quantitative characteristic. To find biomarkers of cell morphological change for quantitatively representing the ratio of morphological changed cell, the mechanism of cell membrane damage driven by typical lipophilic chemicals, such as trichlorophenol (TCP) and perfluorooctanesulphonate (PFOS), was explored. The ratio of morphologically changed cells generally increased with increased TCP or PFOS concentrations, and the level of four major components of phospholipids varied with concentrations of TCP or PFOS, but only the ratio of phosphatidylcholine (PC)/phosphatidylethanolamine (PE) decreased regularly as TCP or PFOS concentrations increased. Analysis of membrane proteins showed that the level of vimentin in normal cell membranes is high, while it decreases or vanishes after TCP exposure. These variations in phospholipid and membrane protein components may result in membrane leakage and variation in rigid structure, which leads to changes in cell morphology. Therefore, the ratio of PC/PE and amount of vimentin may be potential biomarkers for representing the ratio of morphological changed Vero cell introduced by trace lipophilic compounds, thus their composite bio-toxicity.     

Sorptive interactions and binding of δ‐endotoxin protein from bacillus thuringiensis subsp. kurstaki inforest soils

Abstract

The adsorption, desorption and binding of the insecticidal protein from Bacillus thuringiensis subsp. kurstaki (Btk toxin) onto autoclaved sandy and clay loam forest soils were studied at 23°C in a buffer medium (pH 10.2) using the precipitated protein mixture (active + inactive) obtained from a commercial Btk formulation. The active protein in the buffer solution was quantified by ELISA technique. Maximum adsorption of the toxin onto the sandy (301 μg/g) and clay (474 μg/g) loam soils was found to occur after 3 and 4 hours of agitation, respectively. Adsorption of the toxin was higher in the clay loam soil than in sandy loam. Adsorption parameters were calculated using the Freundlich and linear isotherm equations. The K_F and 1/n values for the soils were 1.12 and 1.48 (sandy), and 20.42 and 0.874 (clay), respectively, indicating stronger affinity of the toxin for the clay compared to the sandy loam soil. The linear model showed deviations at higher concentrations, nevertheless using the best fit, K_D and K_OC values were computed for the two soils. For sandy loam, the K_D and K_OC values were 9.38 and 391, respectively; the corresponding values for clay loam were 13.19 and 425, confirming the higher sorption affinity of the toxin for clay loam. The adsorption data did not fit the Langmuir equation because of heterogeneity of the soil surface. Desorption studies showed that more than half of the adsorbed toxic protein remained firmly attached to sandy (162.6 μg/g or 54.5%) and clay (314.0 μg/g or 67.4%) loam soils after six 0.5‐h washes (total 3.0 h wash time). Although the toxin appears to be a non‐leacher, its lateral mobility, soil persistence and biological consequences, including bioavailability of the bound residues, are poorly understood and require further investigation.     

Evolution of Nesting Height in an Endangered Hawaiian Forest Bird in Response to a Non‐Native Predator

Abstract: The majority of bird extinctions since 1800 have occurred on islands, and non‐native predators have been the greatest threat to the persistence of island birds. Island endemic species often lack life‐history traits and behaviors that reduce the probability of predation and they can become evolutionarily trapped if they are unable to adapt, but few studies have examined the ability of island species to respond to novel predators. The greatest threat to the persistence of the Oahu Elepaio (Chasiempis ibidis), an endangered Hawaiian forest bird, is nest predation by non‐native black rats (Rattus rattus). I examined whether Oahu Elepaio nest placement has changed at the individual and population levels in response to rat predation by measuring nest height and determining whether each nest produced offspring from 1996 to 2011. Average height of Oahu Elepaio nests increased 50% over this 16‐year period, from 7.9 m (SE 1.7) to 12.0 m (SE 1.1). There was no net change in height of sequential nests made by individual birds, which means individual elepaios have not learned to place nests higher. Nests ≤3 m off the ground produced offspring less often, and the proportion of such nests declined over time, which suggests that nest‐building behavior has evolved through natural selection by predation. Nest success increased over time, which may increase the probability of long‐term persistence of the species. Rat control may facilitate the evolution of nesting height by slowing the rate of population decline and providing time for this adaptive response to spread through the population.     

Immunoassay of red dyes based on the monoclonal antibody of β-naphthol

The objective of the present study was to develop a multi-analyte immunoassay for the determination of eight red dyes in food samples. Two dye intermediates (2-hydroxy-1-naphthoic acid and 1-amino-2-naphthol) were used as the haptens to produce the monoclonal antibodies. The obtained monoclonal antibodies recognized Sudan 1–4, Para red, Sudan red G, Sudan red B and Acid orange II simultaneously. After evaluation of different antibody/coating antigen combinations, a heterologous indirect competitive enzyme linked immunosorbent assay was developed to determine the eight red dyes in food samples (chili oil, chili powder, tomato sauce, hotpot seasoning). The crossreactivities to the eight analytes were in the range of 61%–79% (with β-naphthol as 100%), and the limits of detection were in the range of 1.3–1.9 ng/mL. The recoveries of the eight analytes from the fortified blank samples were in the range of 84.2%–115% with coefficients of variation lower than 18.3%. Therefore, this method could be used as a rapid and simple tool to detect the residues of the eight red dyes in foods.     

Time aggregation of nitrogen leaching in relation to critical threshold values

Leaching of nitrate contributes to the deterioration of groundwater and can consequently have a negative influence on the quality of our drinking water. Critical threshold values for nitrogen leaching are established to preserve groundwater quality. A critical threshold value for nitrate leaching of 50 mg 1⁻¹ (11.3 mg N 1⁻¹), similar to the drinking water standard, serves as a threshold value for European countries. However, the temporal aggregation scale on which this threshold value should be considered is unknown. A well tested simulation model was used to evaluate the exceedance of the threshold value at different time aggregation levels, ranging from one day till 30 yr. For three different soil structure types within one soil type and a selected fertilisation regime, the aggregated nitrogen leaching over 30 yr was 11.4, 19.2 and 10.6 mg 1⁻¹. Considering an aggregation level of one day, the critical threshold value of 11.3 mg N 1⁻¹ was exceeded 2973, 5801 and 2556 times, respectively, for the three structure types during 30 yr. By considering other time aggregation levels, a clear relation resulted between time aggregation level and the number of time elements during which the critical level was exceeded. Results strongly indicate that a critical threshold value for leaching should include an associated time-aggregation level.