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.     

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.     

DEHP经sirt1/pgc-1a通路诱导的HepG2细胞线粒体损伤效应

为研究邻苯二甲酸二（2-乙基己基）酯（DEHP）是否通过sirt1/pgc-1a通路对细胞产生损伤效应， 通过体外培养HepG2细胞，在1.6， 8， 40， 200， 1000mmol/L DEHP处理24或48h后，采用CCK-8测定细胞活力，ATP试剂盒检测细胞内ATP含量，NO试剂盒检测细胞上清NO含量，ELISA试剂盒检测细胞上清炎症因子TNF-a、IL-6的含量；同时在DEHP作用于HepG2细胞24或48h后，用Western blot检测线粒体调控基因sirt1、pgc-1a，nrf1、tfam的蛋白表达.结果表明：不同浓度的DEHP处理24， 48h后，细胞活力在高剂量都呈显著下降趋势.DEHP能引起ATP含量的显著下降、炎症因子含量的显著提高，但NO含量无显著变化.4种线粒体调控基因的蛋白表达水平sirt1、pgc-1a，nrf1、tfam在24h时无显著变化.而在48h时，随着剂量增加蛋白含量呈上升后下降趋势，并且sirt1在8mmol/L呈显著上升趋势，随后每个蛋白都在1000mmol/L显著下降（P < 0.05）.DEHP能引起HepG2细胞氧化应激，并通过影响sirt1/pgc-1a信号通路表达来影响线粒体生物合成从而造成细胞损伤.     

三环式开发草类资源生产蛋白质食物技术

草类资源分布广、贮量大、成分复杂；至今大部分仍处于自生自灭之中，而人类所需蛋白质营养十分紧张，赖以生存的肉乳蛋品生产所需高蛋白饲料也很缺乏．传统农业、畜牧业、水产业已不能满足这两大需要。作者在科学实验和生产实践的基础上，从工业化生产角度提出以草类为基础原料．经蕈菌－蚯蚓－禽畜３个环节生产蛋白质食物的新工艺路线，对补充上述两大需求的来源提出了一条有效的途径。     

Population dynamics of beech scale (Cryptococcus fagisuga) (Coccina,Pseudococcidae) related to physiological defence reactions of attacked beech trees (Fagus sylvatica)

Summary Changes in the density of beech scale infestation (Cryptococcus fagisuga) of a mature beech stand (Fagus sylvatica) were investigated after gap-cutting in limed and unlimed areas bordering on the gaps and in untreated areas over a 5-year period. Parallelly the concentrations of sucrose, protein amino acids, and procyanidins were examined in the inner and outer bark of non-infested beech trees and beeches infested by beech scale.Irrespective of liming in the areas bordering on the gaps the proportion of beech trees with increasing beech scale infestation was significantly higher, the proportion of beeches with diminishing infestation was significantly lower than in the untreated control areas. Trees with the same infestation tendency occurred in clusters.Physiological defence reactions in the inner and outer bark of the infested beech trees were registrated as an increase in the procyanidin content and a decrease in the protein amino acid content. A change in the outer bark—inner bark—ratio of the infested beech trees showed a transfer of compounds between the inner and outer bark. During the period of observation long-lasting shifts occurred in the pattern of compounds of the bark irrespective of actual infestation intensity.     

Cu和Cd胁迫下接种外生菌根真菌对油松根际耐热蛋白固持重金属能力的影响

铜和镉是我国土壤重金属常见种类,生物固定过量重金属以降低其生理毒害是修复土壤重金属污染的有效方法之一.外生菌根能够通过分泌大量有机物质来降低土壤中重金属的生物有效性.在Cu或Cd胁迫下,外生菌根真菌接种油松幼苗根际分泌的耐热蛋白具有固持重金属的潜力.结果表明,纯培养条件下接种并没有增加耐热蛋白的分泌,但是在Cu或Cd的胁迫培养下,接种幼苗耐热蛋白的分泌量增加.不同Cu浓度处理下,接种油松幼苗总提取耐热蛋白(total thermostable protein,TTP)和易提取耐热蛋白(easily extracted thermostable protein,EETP)的分泌量为未接种油松的2.64~11.79倍;Cd处理下接种油松TTP和EETP的分泌量,分别是未接种油松的1.49~7.56倍.虽然Cu处理下无论是接种还是未接种,根际耐热蛋白和根系细胞中固持的总Cu量显著大于空白对照,但接种显著增加了根际的重金属固持量,其Cu总相对含量是未接种的1.81~2.75倍,且绝大部分重金属固持在耐热蛋白中,使得根际耐热蛋白中的Cu相对含量是根系细胞含量的4.19~43.00倍.Cd处理下也得到了相似的结论.说明接种菌根真菌促进了植物分泌耐热蛋白量,加强了根系对重金属的固持能力,从而达到减少过量重金属的生物有效性,缓解土壤重金属污染的目的.     

含水介质中胞外聚合物的影响因素研究

明确含水介质中微生物胞外聚合物(EPS)生成量的影响因素,是解决含水层生物堵塞的关键问题.本文在现场样品采集和微生物培养的基础上,通过砂柱试验模拟人工回灌过程,系统研究了进水碳、氮、磷浓度、盐度和温度对含水介质中EPS组成和含量的影响.结果表明,砂柱中EPS最主要的成分为多糖和蛋白质,其中多糖占总量的72%~99%;砂柱生物堵塞程度与介质上附着的EPS含量呈正相关关系,当回灌水碳、氮、磷浓度、盐度和温度分别为50 mg·L-1、5 mg·L-1、1 mg·L-1、0.5%和30℃时,微生物EPS生成量最高,砂柱渗透系数降低最显著,堵塞程度最严重.此外,碳、磷浓度、盐度和温度是影响EPS生成量的主要因素,而氮浓度对其影响不大;碳浓度和盐度对EPS中多糖、蛋白质的组成影响较大,而氮、磷以及温度则影响不明显.     

环境因子对脱氯功能念珠藻Nostoc PD-2降解多氯联苯过程中基因表达的影响

以分离筛选自多氯联苯污染水稻田中的脱氯功能蓝藻Nostoc PD-2为材料,两种典型的三氯代多氯联苯PCB28和PCB30为目标污染物,在不同的氮源、碳源和培养温度条件下,研究了脱氯功能藻种Nostoc PD-2中双加氧酶基因和细胞色素b6f复合体铁硫蛋白基因的表达情况及两种基因对脱氯作用的影响.结果显示,以硝酸钠作为氮源时,与氮气氮源组相比,PCB28和PCB30降解组中的两种基因均显著上调表达,双加氧酶基因的上调倍数分别为1.9和5.7倍,铁硫蛋白基因的上调倍数分别为1.1和1.7倍;添加碳酸钠时,与对照相比,双加氧酶基因最高上调了2.2倍,铁硫蛋白基因最高上调了3.4倍;提高温度对双加氧酶基因和铁硫蛋白基因的表达均有促进作用.相较于铁硫蛋白,双加氧酶基因相对表达量与PCB28和PCB30的脱氯百分比之间的相关系数分别为0.872和0.832,表明双加氧酶基因活性与功能藻种PD-2脱氯降解PCBs的相关性更高.本研究结果表明,不同环境因子可引起脱氯功能藻种Nostoc PD-2降解PCBs过程中双加氧酶基因和铁硫蛋白基因的差异表达,添加碳源对降解效果的促进作用最明显,脱氯功能藻种在工程应用中采用优化的环境条件有利于提高降解效率.