五氯苯酚与腐殖酸作用的荧光猝灭效应研究

通过对五氯苯酚与腐殖酸作用的荧光猝灭效应研究,探讨了五氯苯酚与腐殖酸之间的相互作用机理.结果表明,五氯苯酚可以有规律地猝灭腐殖酸的内源荧光,其猝灭机理可认为是五氯苯酚和腐殖酸形成复合物的静态猝灭,五氯苯酚与腐殖酸的结合常数较大,说明腐殖酸能够有效吸附五氯苯酚分子,形成复合物;并获得了不同温度下,五氯苯酚与腐殖酸作用的结合常数和热力学参数.根据所得结果可推断五氯苯酚与腐殖酸的主要作用力为氢键和范德华力.     

基于光谱学和电化学方法的莠去津毒理作用评价

为探索莠去津对DNA的损伤及其毒性作用机制,采用紫外吸收光谱法、荧光光谱法以及电化学方法研究了莠去津与鲱鱼精DNA的相互作用,探讨了二者形成DNA加合物的作用方式.结果表明,莠去津与鲱鱼精DNA作用后,莠去津的紫外光谱呈现减色效应,并有轻微红移现象,而其荧光光谱强度明显增强;循环伏安法显示莠去津与DNA作用能引起莠去津还原电位正移,峰电流减小.以上实验结果表明,莠去津平面分子能够嵌插到DNA双螺旋链中,形成较稳定的加合物.采用电化学方法测得莠去津与鲱鱼精DNA的结合比为1:1,结合常数为1.2×105.该研究为莠去津毒理作用评价提供了一种简便的方法。     

采用光谱学技术分析邻苯二甲酸酯与DNA互作机制

邻苯二甲酸酯(PAEs)作为增塑剂被广泛使用,易渗透到环境中产生环境毒性,多种PAEs已被美国环境保护署和中国环境监测中心列为优先控制污染物,其生态毒理学机制被广泛关注。本试验以小牛胸腺DNA(ct DNA)为供试材料,旨在通过光谱学技术探究PAEs与DNA的相互作用机制。在紫外可见光谱试验中,随着PAEs浓度的增加,DNA紫外光谱出现增色效应,证明DNA的部分碱基对被PAEs破坏,且PAEs与DNA通过嵌插作用结合。荧光光谱试验中,随着PAEs浓度的增加,EBct DNA体系荧光强度逐渐降低,邻苯二甲酸二甲酯(DMP)、邻苯二甲酸二乙酯(DEP)和邻苯二甲酸二丁酯(DBP)的最大荧光抑制率分别为55%、50%和36%;KI荧光猝灭试验中,在加入DNA之后KI对DMP、DEP和DBP的荧光猝灭常数Ksv分别从7.992(R2=0.9970)、10.270(R2=0.9960)和13.52(R2=0.9806)降低到6.721(R2=0.9963)、7.047(R2=0.9599)和11.03(R2=0.9803),荧光猝灭常数均降低,实验结果证明PAEs与DNA通过嵌插作用结合。盐离子试验中,随着Na Cl浓度的增加,PAEsct DNA的荧光强度没有发生变化。试验结果排除了PAEs与DNA分子之间沟槽作用的可能,确定了其结合方式为嵌插作用,随着PAEs侧链的逐渐增长,嵌插作用逐渐减弱。     

环糊精对腐植酸-芘、蒽相互作用影响初探

利用吸收光谱、荧光猝灭法、圆二色谱法及芘荧光探针法,探讨了环糊精(CDs)对腐植酸(HA)与以芘为代表的多环芳烃(PAHs)相互作用的影响.β-CD与HA的吸收光谱和荧光光谱测定结果表明:β-CD能够包络HA的芳环部分;荧光猝灭法和圆二色谱法测定结果表明:HA能够与被β-CD空腔包络的芘分子发生相互作用,CDs和HA之间存在与芘相互作用的竞争;用荧光猝灭法测定了四种不同CDs存在下HA和芘的结合常数,并用芘为荧光探针进行测定,结果表明:CDs浓度较低时,HA对芘的作用占优势;反之,CDs占优势.与其它三种CDs不同,γ-CD存在下HA与芘的结合常数随其浓度的升高一直增大.     

渗滤液中溶解性荧光有机物与铅的作用

采用荧光光谱和猝灭滴定实验,结合二维相关分析和区域体积积分分析,研究了生活垃圾渗滤液不同处理阶段样品中溶解性荧光有机物(DFOM)络合Pb(Ⅱ)的特性.结果显示:渗滤液中无机阴离子对Pb(Ⅱ)猝灭DFOM荧光影响较大,它们的存在致使Pb(Ⅱ)对DFOM的荧光猝灭效果不明显.在渗滤液DFOM的各种荧光基团中,Pb(Ⅱ)对类蛋白类物质的荧光猝灭效率高,而对类腐殖质物质的荧光猝灭效率低,这种猝灭特征不随渗滤液处理进行而改变;但Pb(Ⅱ)对调节池和厌氧处理中可溶性微生物降解副产物的荧光猝灭效率低而对氧化沟和MBR出水中DFOM的荧光猝灭效率高.分析结果表明,渗滤液DFOM中类蛋白物质络合Pb(Ⅱ)能力大于类腐殖质物质,经好氧生物处理后,可溶性微生物降解副产物络合Pb(Ⅱ)的能力有所提高.     

4-硝基苯胺与人血清白蛋白相互作用的研究

通过荧光和紫外光谱法研究了4-硝基苯胺与人血清白蛋白（HSA）的作用.结果表明,4-硝基苯胺对HSA的内源荧光具有强烈的猝灭作用.猝灭机理为静态猝灭,同时伴随有非辐射能量转移的发生.根据双对数方程计算其结合常数和结合位点数.确定4-硝基苯胺与HSA有一类结合部位.根据热力学参数得出4-硝基苯胺与HSA之间的主要作用力为氢键和疏水作用力.同步荧光的结果表明,作用点位靠近色氨酸,并且使色氨酸的疏水环境增强.     

荧光光谱法研究草甘膦与腐殖酸的相互作用

应用荧光光谱法研究了草甘膦(PMG)与腐殖酸(HA)的相互作用.荧光猝灭实验表明,PMG对HA的荧光产生静态猝灭;通过对不同温度下反应热力学参数的计算,发现二者相互间作用力主要是氢键和范德华力.受到PMG作用的影响,HA的同步荧光光谱出现了蓝移现象,初步说明HA在作用后共轭度降低.采用平行因子分析技术对加入PMG前后的HA三维荧光光谱进行解析,发现加入PMG后HA荧光组分不仅出现在区域Ⅴ(难降解有机物荧光区)中,而且有部分出现在了区域Ⅲ(可降解有机物荧光区)中,总荧光强度明显减弱,而且组分C3、C4有明显的蓝移现象.三维荧光光谱分析验证了荧光猝灭及同步荧光的研究结果,并进一步表明,PMG与HA的相互作用使HA共轭度降低,并且可能有可降解物质生成.     

掺氮碳量子点的制备及对偶氮类色素的荧光检测

以苯酚为碳源、乙二胺为氮源,采用一步水热法制备了水溶性的掺氮碳量子点(N-CQDs),用于偶氮类色素的荧光检测.首先对N-CQDs的形貌、组成和荧光特性进行了表征,然后研究了色素柠檬黄和胭脂红对N-CQDs体系显著的荧光猝灭效应,优化了实验条件.结果表明,该氮掺杂方法当苯酚-乙二胺摩尔比接近1∶1时,在N-CQDs表面可形成大量强供电子基团,有效提高了其荧光效率;柠檬黄、胭脂红对N-CQDs的荧光猝灭在5min内达到平衡,在磷酸盐缓冲溶液(pH=7.5)的条件下荧光猝灭效率最高,且均为静态猝灭.最佳条件下,柠檬黄浓度[Q]在0—80 nmol·L~(-1)范围内与N-CQDs体系的荧光猝灭程度(lgF_0/F)呈现良好的线性关系:lg F_0/F=0.0047[Q]-0.015,R~2=0.994,检出限为8.20 nmol·L~(-1);胭脂红浓度[Q]在0—40 nmol·L~(-1)范围内与N-CQDs体系的荧光猝灭程度(lg F_0/F)也呈现良好的线性关系:lg F_0/F=0.0252[Q]-0.0052,R~2=0.996,检出限为1.54 nmol·L~(-1).本研究为简便、快捷、灵敏地检测废水中偶氮类色素柠檬黄和胭脂红提供了新的方法.     

结合高分辨质谱法、荧光光谱法及分子对接研究全氟化合物与白蛋白的相互作用

本文结合电喷雾离子源-四极杆串联时间飞行高分辨质谱法(ESI-QTOF HRMS)、荧光光谱法(FL)以及分子对接(MD)实验手段,研究了全氟辛酸(PFOA)、全氟十二酸(PFDoA)与牛血清白蛋白(BSA)的相互作用机制.首先,采用HRMS方法检测到了PFOA、PFDoA与BSA结合物的分子量信息,证实了这两种污染物与BSA能形成稳定复合物;利用荧光光谱法证实了两种污染物对BSA的荧光猝灭为静态猝灭,进一步验证了PFOA、PFDoA与BSA之间复合物的形成,同时计算了两种污染物对BSA的结合常数和结合位点数,得出PFDoA与BSA的结合常数更高的结论,这一实验结果也与其他研究工作结果互为印证,即全氟化合物的C—F链长对其与生物分子的分配常数的正比关系.另外,使用分子对接研究手段进一步验证了PFOA、PFDoA与BSA的3个结合位点之间均存在相互作用,两种污染物的极性端与BSA氨基酸残基直接形成氢键,疏水端则与非极性残基有疏水相互作用,氢键作用与疏水作用共同促进PFCs有机污染物与蛋白质的相互结合.     

2-脱氧-3-磷酸胞嘧啶-苯醌加合物的确定

用~(32)P后标记法,HPLC及PEI·TLC同谱层析等方法对DNA-苯醌(BQ)加合物进行了研究,并对其中一个主要加合物进行了碱基定位,确证该斑点系BQ与分子DNA上的2′-脱氧-3′-磷酸胞嘧啶作用形成的加合物(dC-BQ加合物)。     

应用紫外及荧光光谱法研究三种醛类污染物对牛腺DNA的损伤作用

应用紫外光谱移动法和溴乙锭荧光法对三种醛类化合物与牛腺ＤＮＡ的加合以及它们引起的ＤＮＡ－ＤＮＡ交联进行了检测。结果表明,甲醛和乙醛能够与ＤＮＡ形成共价加合物,与四种脱氧核苷加合顺序分别为ｄＧ〉ｄＣ〉ｄＡ〉ｄＴ和ｄＡ〉ｄＧ〉ｄＣ〉ｄＴ,丙烯醛与ＤＮＡ的加合不能用紫外光谱移动法测定。甲醛能够显著地引起ＤＮＡ－ＤＮＡ的交联;乙醛也有这种作用,但不显著;丙烯醛的交联作用未测出,可能是由于羰基碳正电性较弱,     

涕灭威及其有毒代谢产物对DNA潜在损伤研究

采用生物遗传监测系统,研究了涕灭威及其两个有毒代谢产物(涕灭威亚砜,涕灭威砜)对蚕豆及大蒜根尖细胞微核率的影响,以及对小牛胸腺DNA的加合作用．结果表明,两种植物根尖细胞微核试验测得的高浓度剂量诱导的微核率与对照组相比有显著性差异(p<0．05),说明涕灭威、涕灭威亚砜及涕灭威砜可能对蚕豆及大蒜根尖细胞有致突变性．并且配对资料差异的显著性检验表明,蚕豆根尖微核试验的敏感性明显高于大蒜根尖微核试验的敏感性．涕灭威及其两个有毒代谢产物与小牛胸腺DNA结合引起吸收光谱波峰的移位,而且存在着剂量关系,说明涕灭威、涕灭威亚砜及涕灭威砜可能对小牛胸腺DNA造成加合作用．     

Evidence for DNA-diquat interaction and cytotoxicity in in vitro rat cells

Over 98% of sprayed insecticides and 95% of herbicides reach non-target species in air, soil, and water. Numerous studies have reported that pesticide residues can cause acute and chronic toxicity. Pesticide residues can be carcinogenic, mutagenic, and immunotoxic. There are actually too few studies that bridge the disciplines of chemobioanalysis and environmental toxicology. Here, we assessed the cytotoxicity of a bipyridilium herbicide diquat in rat adrenal pheochromocytoma cells (PC12). Our results show that diquat caused the decrease in cell viability with a lethal concentration 50 (LC₅₀) of 1.4 × 10⁻⁵ mol/L. This cytotoxicity may result from diquat-induced apoptosis, characterized by nuclear fragmentation and chromatin condensation by Hoechst 33324 staining. To explore the possible mechanisms, the interaction between herbicide diquat and calf thymus DNA (ctDNA) was further investigated using fluorescence quenching. The detection of static quenching showed that diquat was linked with ctDNA by electrostatic interaction with a binding constant of 9.288 × 10⁴ L/mol. This is the first study on the interaction of DNA with herbicide diquat by fluorometric method as well as on the evaluation of cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Hoechst staining. Given the widespread use of synthetic pesticides, the data would be valuable for the risk assessment of pesticide residues.     

荧光光谱法研究木栓酮与牛血清白蛋白的相互作用

为了解三萜类化合物在体内的结合转运机制,采用荧光光谱法结合紫外吸收光谱研究从中药爵床中分离的有效成分木栓酮(Friedelin,FDL)与牛血清白蛋白(Bovine serum albumin,BSA)的结合反应.通过研究木栓酮对牛血清白蛋白内源性荧光的淬灭作用,发现木栓酮可以与牛血清白蛋白相互结合,且其作用是一个静态猝灭的过程.通过计算木栓酮与牛血清白蛋白的结合常数和结合位点,发现在生理弱碱性溶液中二者具有较强的结合作用,且以1:1结合.根据热力学参数推测木栓酮与牛血清白蛋白的作用力类型为氢键和范德华力.根据F rster非辐射能量转移机理,计算出木栓酮与牛血清白蛋白结合距离为2.86 nm.研究表明木栓酮在体内可以通过血清白蛋白进行结合和转运.     

Effect of the chemical composition of organic extracts from environmental and industrial atmospheric samples on the genotoxicity of polycyclic aromatic hydrocarbons mixtures

Particulate organic matter (PM) present in the atmosphere is a complex mixture of chemicals like polycyclic aromatic hydrocarbons (PAH) that may exert adverse health effects including respiratory and cardiovascular disturbances and cancer. In this study, airborne samples from environmental or industrial areas exhibiting different physicochemical composition were compared for their capacities to induce DNA damage in human hepatocytes HepG2. DNA strand breaks and DNA adducts formed by benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE), the most reactive metabolite of the carcinogenic benzo[a]pyrene (B[a]P), were measured with the comet assay and by high-performance liquid chromatography (HPLC)/mass spectrometry, respectively. Cells were exposed to organic matter extracted from PM. Experiments were performed either at a constant concentration of B[a]P or at concentrations corresponding to fixed air volumes. Results show that industrial extracts tend to produce more benzo[a]pyrene diol epoxide-N ²-2′-deoxyguanosine (BPDE-N ²-dGuo) DNA adducts than strand breaks, whereas the opposite was observed with environmental extracts. The chemical composition of the extracts significantly impacts the nature and levels of DNA damage. The amount of B[a]P and interaction with other contaminants in the extracts need to be considered to explain the formation of DNA damage. These results emphasize the use of in vitro tests as promising and complementary tools to widely used toxic equivalent factor (TEF) approach in order to assess health hazards related to chemical exposure of the general population.     

Spectrophotometric study of DNA interactions with ftorafur and its elementoorganic derivatives

Ftorafur is an antimetabolite antitumour drug successfully used for treatment of various tumours. It is generally accepted that ftorafur is converted to 5-fluoruracil. However, some data indicate direct interactions of the compound with DNA. To test this hypothesis we have performed spectrophotometric study of DNA interactions of ftorafur and some of its elementoorganic derivatives with DNA. UV-VIS spectra of the tested compounds were recorded in absence and presence of increasing amounts of DNA. DNA caused a hypochromic effect in spectra of ftorafur, similar, but weaker effect was observed in 5-fluoruracil spectra. Trimethylgermyl derivative of ftorafur manifested a higher DNA-binding capacity compared to ftorafur. To reveal possible mechanism of interaction between the tested nucleosides and DNA ethidium bromide extrusion experiments were performed. It was shown that the compounds did not compete with EBr for intercalation. Fourier transformation infrared spectroscopy analyses revealed decrease in intensity of several bands in spectra of ftorafur, its trimethylgermyl derivative and fluorouracil in presence of DNA indicating again interactions between the compounds and DNA. Docking experiments reveal interactions of the tested nucleosides with the DNA minor groove. Thus ftorafur is capable to interact directly with DNA; further modifications of the molecule enhance this capacity.     

西维因对雄性罗非鱼(GIFT Oreochromis niloticus)内分泌干扰效应的研究

在实验室条件下研究了西维因对罗非鱼的急性毒性以及在不同暴露浓度下的西维因(0.064,0.8和5mg·L-1)对雄性罗非鱼性腺系数、血清雌二醇、睾酮含量的影响以及对卵黄蛋白原的诱导效应,以期对其环境雌激素效应研究有所贡献。结果表明:西维因对罗非鱼的24、48、72和96h的LC50分别为30.21、25.16、19.76和17.86mg·L-1。长期暴露于低剂量的西维因中,会使雄性罗非鱼精巢系数降低;对其血清雌二醇的合成具有诱导作用,当浓度为0.8mg·L-1时,诱导作用最强,此时血清雌二醇的含量为218.83pg·mL-1;卵黄蛋白原的含量随着西维因浓度的升高,逐渐增多,当处理浓度为5mg·L-1时,达到最大为393.01μg·L-1;随着西维因浓度的升高,睾酮含量逐渐降低,西维因的浓度为5mg·L-1时,睾酮含量达到10.58mol·L-1,差异极显著(p<0.01),这可能是由于西维因对雄性罗非鱼具有雌激素效应。     

Carotenoid-based ornamentation as a dynamic but consistent individual trait

Carotenoid-based ornaments act as signals of quality in many animal species. In contrast to feathers, which are relatively stable structures, carotenoid-pigmented integuments (e.g. bills, lores, tarsi) can change colour rapidly and may better reflect changes in physiological condition. I studied the seasonal variations in plasma carotenoids in red-legged partridges (Alectoris rufa) kept on a constant diet and free of intestinal parasites. Furthermore, I analyzed whether seasonal changes in circulating carotenoids were mirrored by the carotenoid-based coloration of eye rings and bill of this species. Plasma carotenoids showed seasonal variation, with higher levels coinciding with the end of the mating and the start of the laying season. Eye ring pigmentation was related to plasma carotenoid levels, and changes in bill hue (but not changes in UV or red bill chroma) mirrored the variation in plasma carotenoids during the breeding season. Despite the seasonal variation, individual differences in eye ring pigmentation and bill hue, UV and red chroma were consistent throughout the breeding season. Similarly, individual differences in eye ring pigmentation and bill hue and red chroma remained consistent between consecutive years. These results suggest that carotenoid based integumentary colorations act as dynamic traits that accurately reflect the carotenoid-status of individuals, thus reliably indicating consistent differences in individual quality. Furthermore, variability in signal expression appears to have a relevant genetic/phenotypic basis independently of environmental conditions. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.