铅对人Bel-7402肝癌细胞毒性及EGFP基因DNA甲基化的影响

以体外培养人Bel-7402肝癌细胞为模型,研究铅的3种常见化合物氯化铅(Pb Cl2)、乙酸铅(Pb(CH3COO)2)、硝酸铅(Pb(NO3)2)的细胞毒性和去甲基化表观遗传毒性。应用MTS方法检测细胞的存活率,以前期研究建立的评价方法评价铅化合物的去甲基化表观遗传毒性。结果显示,Pb Cl2、Pb(CH3COO)2、Pb(NO3)2均会抑制Bel-7402细胞的增值,计算求得Pb Cl2、Pb(CH3COO)2、Pb(NO3)2相应的50%细胞存活浓度(IC50)值分别为2 524μmol·L~(-1)、1 977μmol·L~(-1)、1 899μmol·L~(-1);80%细胞存活浓度(IC80)值分别为264μmol·L~(-1)、221μmol·L~(-1)、281μmol·L~(-1),通过对3种染毒物不同染毒浓度的细胞存活率进行随机区组设计的方差分析显示3种化合物间的差异无统计学意义(F=0.11;P=0.897)。去甲基化表观遗传毒性检测结果显示,Pb Cl2、Pb(CH3COO)2、Pb(NO3)2均可观察到明显的去甲基化表观遗传毒性,其相对于5-氮杂-2-脱氧胞苷(5-Aza-Cd R)的去甲基化表观遗传毒性当量分别为2.82E-03、1.50E-03、5.09E-04,三者间也无显著性差异。结果表明,铅化合物会使Bel-7402细胞的细胞存活率和转染进细胞的质粒上增强型绿色荧光蛋白基因启动子的DNA甲基化水平下降。     

PFOS对星形胶质细胞表观遗传调控作用初探

脑源性神经营养因子(BDNF)甲基化在全氟辛烷磺酸(PFOS)神经毒性中的作用已证实,但表观遗传修饰中其他调控因子在PFOS对星形胶质细胞毒性中的影响仍有待探索。本文以大鼠原代星形胶质细胞为体外生物体系,建立24 h PFOS(0、25、50和100μmol·L~(-1))暴露模型,通过观察PFOS暴露对星形胶质细胞表观遗传调控主要分子DNA甲基化酶(DNMTs)、组蛋白去乙酰化酶(HDACs)和小泛素化修饰物(SUMOs)的影响,初步明确表观遗传调控机制参与PFOS神经毒性作用。采用Hoechst 33258检测细胞凋亡,利用ELISA试剂盒检测HDACs含量,以实时荧光定量PCR考察DNMTs、HDACs和SUMOs基因表达。结果显示,星形胶质细胞暴露于一定浓度PFOS(≥25μmol·L~(-1))时产生凋亡现象(P0.05),HDACs含量升高(P0.05),且DNMT1、HDAC1/2/4与SUMO-1的基因表达显著升高(P0.05);而当PFOS浓度高于50μmol·L~(-1)时,可显著诱导DNMT3A、SUMO-2的基因表达(P0.05); DNMT3B在PFOS≥25μmol·L~(-1)时,其基因表达具有升高趋势,但不具统计学显著性(P0.05)。结果表明,PFOS可以影响星形胶质细胞的表观遗传修饰;表观遗传修饰可能是PFOS神经毒性作用机制之一。     

环境污染物诱导神经毒性的表观遗传机制研究进展

表观遗传修饰与神经系统功能密切相关,其在环境污染物暴露致神经毒性中的作用机制已引起广泛关注。本文综述了重金属、有机污染物和空气颗粒物等典型环境污染物对人体和模式生物表观遗传修饰(DNA甲基化、组蛋白修饰和ncRNA等)和神经系统功能的影响,指出环境污染物可直接或(通过引起氧化胁迫)间接改变表观遗传修饰状态,导致相关基因表达失调,从而诱导一系列神经毒性,并提出当前研究存在的局限性。建议未来针对污染物的神经毒性机制研究,应着重关注组蛋白修饰和ncRNA以及不同类型表观遗传修饰之间的交互作用;同时,环境污染物复合暴露导致神经毒性的表观遗传机制有待深入研究;如何从表观遗传角度解释环境污染物诱导神经毒性的年龄易感性及性别特异性也值得进一步探讨。     

废弃农药厂污染场地土壤浸出液的急性毒性和遗传毒性筛查

污染土地再开发是城市土地可持续利用的关键。为进行废弃农药厂的生态风险评价,采集常州市某废弃农药厂污染场地6个不同区域的土壤样品(编号为S1、S2、S3、S4、S5和S6),选取发光细菌、大型溞和蚕豆根尖细胞为试验生物,运用成组生物毒性试验对6个土壤样品的浸出液进行生物毒性检测。结果表明:6个土壤样品的浸出液都具有急性毒性和遗传毒性;发光菌和大型溞急性毒性试验表明,S3和S5的土壤浸出液毒性最高,S6的土壤浸出液毒性最低;蚕豆根尖微核遗传毒性试验表明S3和S5的土壤浸出液毒性最高,S4和S6的土壤浸出液毒性最低。蚕豆根尖微核遗传毒性试验结果与发光菌和大型溞急性毒性效应基本相同,表明成组生物毒性测试法可以用于污染土壤浸出液的毒性评价。污染物质和毒性检测结果的相关性分析结果表明,土壤中所含污染物种类和浓度已经对土壤毒性效应产生影响,结合化学分析与生物毒性检测可为污染场地进行综合评价与风险评估提供依据。     

利用改进的SOS/umu方法检测水处理过程中污染物的遗传毒性效应

SOS/umu遗传毒性测试方法被广泛应用于化合物和复杂混合物遗传毒性的评价,但是目前已报道方法的实验周期长达17 h,给实验操作带来了不便,因此亟需对常规方法进行改进。建立了测试周期可小于8 h(2 h预培养,4.5 h暴露培养) 的SOS/umu快速测定方法之后,应用改进后的方法对5种典型遗传毒性物质, 4-硝基喹啉1-氧化物(4-NQO)、丝裂霉素C(MMC)、甲磺酸甲酯(MMS)、2-氨基蒽(2-AA)和苯并芘(BaP)进行测定,结果表明,改进后的SOS/umu方法对4-NQO、MMC、MMS、2-AA和BaP的检测限分别为0.013±0.0031、0.031±0.0028、229.18±60.51、2.29±1.22和1.28±0.0698 μmol·L^-1,优于或者相当于报道方法。采用经典方法和改进后的方法对全国两个地区6个环境水样的遗传毒性进行测定,结果表明,两种方法检测出的6个环境水样的遗传毒性强度无显著性差异。另外,将研究方法应用于北方某市甲乙两个水厂原水、出厂水和管网水的遗传毒性评价,结果表明,两个水厂的原水、出厂水和管网水均表现出一定的直接遗传毒性,对应的4-NQO当量浓度(TEQ4-NQO)在0.0012 μg·L^-1至0.0129 μg·L^-1之间;加入鼠肝微粒体酶系统(S9)后,仅有乙水厂的管网水检测出极其微弱的遗传毒性效应。以上研究结果表明,改进后的SOS/umu快速测定方法能够实现在短时间（8h）内完成环境化合物及环境样品的遗传毒性检测,为筛选和评价化合物和复杂环境样品的遗传毒性提供了更为快捷的手段。     

基于发光细菌的细颗粒物(PM_(2.5))可溶性提取液综合生物效应测试方法及其应用

为科学评估PM_(2.5)对生物体综合生物效应,研究建立了利用费氏弧菌检测PM_(2.5)水溶性提取液的毒性测试方法,确立了PM_(2.5)样品提取液发光细菌毒性测试实验质量控制办法。对春节烟花爆竹燃放和沙尘污染过程的PM_(2.5)实样测试表明:烟花爆竹燃放期间的PM_(2.5)样品提取液发光抑制率值与微量金属元素等有毒有害组分浓度显著相关;沙尘污染期间的PM_(2.5)样本提取液中地壳元素浓度和发光抑制率值显著不相关。     

荧光光谱法快速检测土壤中荧光烃类污染物

探讨了正己烷、石油醚及二氯甲烷∶正己烷(1∶2,体积比)混和液三种溶剂浸泡或超声波萃取与荧光光谱相结合测定土壤中荧光烃类污染物的方法,并为了验证方法有效性,对典型煤矿区93个土壤样品中的荧光烃类进行了快速检测。结果表明,采用二氯甲烷∶正己烷(1∶2,体积比)混和液超声波萃取土壤中荧光烃类,耗时短,萃取能力较强。对于具有大量土壤样品的研究区域,超声波萃取与荧光光谱法相结合可大大缩短土壤样品中总荧光烃类污染物的检测时间,进而可快速获知某一研究区内土壤环境中荧光烃类污染物的空间分异规律及相对污染程度,具有一定的应用价值。     

荧光假单胞菌(Pseudomonas fluorescens)工程菌株田间残留和扩散的追踪检测

分别在北京和江苏省连云港市对携带Btcry基因的荧光假单胞菌工程菌进行了田间残留与扩散的追踪检测 .对越冬前后的试验地和保护地土壤样品进行抗生素抗性平板分离 ,能检测到极少量的具有与出发菌株相同抗性的荧光假单胞菌菌落 ,但没有发现工程菌株的残留与扩散 .对江苏试验地样品还进行了工程菌株质粒卡那霉素和壮观霉素抗性标记基因的抗性菌落分离 ,绝大多数样品中都能分离到包括荧光假单胞菌在内的抗性菌落 ,土样中菌密度n(cfu) =10 4 ～ 10 5g-1,但进行Btcry基因PCR -RFLP检测时没有从样品中得到特异扩增产物 .研究结果表明 :工程菌株抗性标记基因在自然界广泛存在 ,工程菌在株环境中没有残留和扩散 ,具有良好的生物安全性 .表 4参 8     

Cl-BDE-208和BDE-209诱导LINE-1基因低甲基化和ESR1基因高甲基化

十溴联苯醚(BDE-209)是使用最多的一种多溴阻燃剂,由于具有高亲脂性和低挥发性,容易蓄积在生物体内,在人体血清、母乳、肝脏等组织中均有检出;其环境脱溴产物九溴一氯联苯醚(Cl-BDE-208)也在血清样品和环境样品中被频频检出,而目前对这种化合物的毒理学研究,特别是对人体早期健康效应的研究较少。为了评估Cl-BDE-208和BDE-209的早期健康效应,以人乳腺癌细胞MCF-7为模型,环境相关浓度(0.375~3μmol·L-1)为暴露浓度,检测Cl-BDE-208和BDE-209对重复序列LINE-1、全基因组DNA甲基化(GDM)和雌激素受体基因(包括ESR1和ESR2)甲基化水平的改变。结果表明,Cl-BDE-208和BDE-209都可能通过降低DNA甲基化转移酶(DNMTs)活性而诱导了LINE-1和GDM的低甲基化,8-羟基脱氧鸟苷(8-OHd G)水平的升高可能参与了BDE-209引起的低甲基化过程;同时通过促进雌激素受体α(ESR1)基因的高甲基化而显著下调了ESR1基因的表达,而对雌激素受体β(ESR2)基因甲基化水平并没有显著影响。Cl-BDE-208和BDE-209都可能通过改变DNA甲基化水平而影响人体的早期健康效应,同时ESR1基因的高甲基化可能是Cl-BDE-208和BDE-209引起内分泌毒性的机制之一。     

土壤样品中微生物活性的荧光分析方法

以土壤为供试样品,基于荧光素二乙酸酯(FDA)水解反应产生的荧光素含量与土壤中微生物活性呈比例的现象,探讨了荧光光度法测定土样中微生物活性的可行性,提出采用荧光光度法测定环境样品中微生物活性的新方法.对测定所涉及的物理参数和化学参数进行了优化.在最佳实验条件下,方法的线性响应范围为荧光素含量2—2000#g.L-1,线性相关系数(r)=0.9971,方法的检出限(3σ)为3.1×10-2#g.L-1,方法的相对标准偏差(RSD)为2.8%(n=5),对4个土壤样品荧光素含量测定的回收率为90.6%—104%.所建立的方法具有简单快速、灵敏度高、线性范围宽、试样用量少等优点,可用于土壤、水等环境样品中微生物活性的测定.     

Achieving Open Access to Conservation Science

Conservation science is a crisis discipline in which the results of scientific enquiry must be made available quickly to those implementing management. We assessed the extent to which scientific research published since the year 2000 in 20 conservation science journals is publicly available. Of the 19,207 papers published, 1,667 (8.68%) are freely downloadable from an official repository. Moreover, only 938 papers (4.88%) meet the standard definition of open access in which material can be freely reused providing attribution to the authors is given. This compares poorly with a comparable set of 20 evolutionary biology journals, where 31.93% of papers are freely downloadable and 7.49% are open access. Seventeen of the 20 conservation journals offer an open access option, but fewer than 5% of the papers are available through open access. The cost of accessing the full body of conservation science runs into tens of thousands of dollars per year for institutional subscribers, and many conservation practitioners cannot access pay‐per‐view science through their workplace. However, important initiatives such as Research4Life are making science available to organizations in developing countries. We urge authors of conservation science to pay for open access on a per‐article basis or to choose publication in open access journals, taking care to ensure the license allows reuse for any purpose providing attribution is given. Currently, it would cost $51 million to make all conservation science published since 2000 freely available by paying the open access fees currently levied to authors. Publishers of conservation journals might consider more cost effective models for open access and conservation‐oriented organizations running journals could consider a broader range of options for open access to nonmembers such as sponsorship of open access via membership fees. Obtención de Acceso Abierto a la Ciencia de la Conservación     

Rapid,recurring, structured survey versus bioblitz for generating biodiversity data and analysis with a multispecies abundance model

A bioblitz inexpensively and quickly generates biodiversity data, but bioblitzes are often conducted with haphazard, unreplicated sampling. Results tend to be taxonomically, geographically, or temporally biased, lack metadata, and consist of lists of observed taxa that do not enable further analyses or correction for imperfect detection. A rapid, recurring, structured survey (RRSS) uses a structured sampling design and temporal and spatial replication to survey randomly selected sites on a conservation property. We participated in a loosely structured bioblitz and a subsequent RRSS at Big Canoe Creek Nature Preserve in Springville (St. Clair County), Alabama (USA) to compare observed richness derived from the 2 survey approaches. The RRSS data structure enabled us to fit models that accounted for imperfect detection to estimate abundances, occupancy probabilities, and habitat associations. The loosely structured bioblitz data could not be used in such models. We present a new integrated multispecies abundance model that we applied to avian RRSS data. Our model extension enables estimation for the community, employs data augmentation to estimate the number of undetected species, and incorporates covariates. The RRSS generated a more comprehensive and less biased list of observed taxonomic richness than the loosely structured bioblitz (e.g., 73 vs. 45 bird species and 104 vs. 63 insect families from the RRSS vs. loosely structured bioblitz, respectively). Models fit to the RRSS data identified seasonal patterns in avian community composition and allowed for estimation of habitat–occupancy relationships for insect taxa. The RRSS protocol has potential for broad transferability as a standardized, quick, and inexpensive way to inventory biodiversity and estimate ecological parameters while providing an outreach opportunity.     

Compliance with and ecosystem function of biodiversity offsets in North American and European freshwaters

Land-use change via human development is a major driver of biodiversity loss. To reduce these impacts, billions of dollars are spent on biodiversity offsets. However, studies evaluating offset project effectiveness that examine components such as the overall compliance and function of projects remain rare. We reviewed 577 offsetting projects in freshwater ecosystems that included the metrics project size, type of aquatic system (e.g., wetland and creek), offsetting measure (e.g., enhancement, restoration, and creation), and an assessment of the projects’ compliance and functional success. Project information was obtained from scientific and government databases and gray literature. Despite considerable investment in offsetting projects, crucial problems persisted. Although compliance and function were related to each other, a high level of compliance did not guarantee a high degree of function. However, large projects relative to area had better function than small projects. Function improved when projects targeted productivity or specific ecosystem features and when multiple complementary management targets were in place. Restorative measures were more likely to achieve targets than creating entirely new ecosystems. Altogether the relationships we found highlight specific ecological processes that may help improve offsetting outcomes.     

Some polyphagous Homoptera gain protection from their natural enemies by feeding on the toxic plantsSpartium junceum andErythrina corallodendrum (Leguminosae)

Summary Interactions were studied among alkaloid-containing legumes (Erythrina corallodendrum andSpartium junceum) and non-toxic plants (Citrus sinensis, Cucurbita moschata andEuphorbia tirucalli), several polyphagous homopterans,Aphis craccivora (Aphididae),Icerya purchasi, I. aegyptiaca (Margarodidae),Lepidosaphes ulmi (Diaspididae) andPlanococcus citri (Pseudococcidae), and some major natural enemies of these homopterans. Significant reductions in survival due to negative effects of alkaloid containing as compared with non-alkaloidal plants were recorded for the predatorsRodolia cardinalis andChilocorus bipustulatus, but not forCryptolaemus montrouzieri (Coleoptera: Coccinellidae),Chrysoperla carnea (Neuroptera: Chrysopidae) andSympherobius sanctus (Neuroptera: Sympherobiidae). The development time of the larvae or pupae ofR. cardinalis, C. carnea andS. sanctus was longer on the toxic plants than on the non-toxic ones. The percentage of parasitism ofA. craccivora collected from the non-alkaloidal plantsVicia palaestina andMelilotus albus was much higher than that onS. junceum. The parasitoid complexes ofA. craccivora differed between both plant groups. The nutritive value of honeydew ofI. purchasi andA. craccivora, as expressed by the life span ofEncyrtus infelix (Hymenoptera: Encyrtidae) adults, was also investigated. Life spans were significantly longer when the wasps fed on honeydew produced on non-alkaloidal plants (C. sinensis andPittosporum tobira) than on alkaloid containing plants whenI. purchasi — but notA. craccivora — was the producer. It is suggested that the chemical defense ofE. corallodendrum andS. junceum is exploited by polyphagous phytophages to reduce predation. In nature, population growth and density of four of the investigated homopterans are conspicuously high when they developed on the alkaloid containing plant species, and very low on non-alkaloid plants. The efficiency of their natural enemies may be reduced by sequestration of alkaloids (or other toxic plant compounds) or their transfer into excreted honeydew. Therefore it is assumed that a generalist phytophagous homopteran may be protected from its natural enemies, although at different rates of efficiency, if it can safely sequester the host allelochemical when it develops on toxic species within its host range.     

A palaeo-hydrogeological model for arsenic contamination in southern and south-east Asia

An argument is presented in which areas of natural arsenic contamination of modern groundwaters throughout Asia have a common origin. Arsenic originally accumulated in oceanic ferro-manganoan sediments of the eastern Palaeo-Tethys. This was further concentrated through oceanic crustal extinction in what later became the south-east Chinese accreted mineralised terrain. Proto-Himalayan uplift of this area created the palaeo-drainage systems of the Ganges – Brahmaputra, Irrawaddy, Mekong, and Red Rivers, with consequent headwater erosion of arsenic-rich sediments. Their downstream deposition as immature and easily redistributed Neogene sandstones, silts, and iron-rich clays has created secondary and tertiary reservoirs of adsorbed and authigenic arsenic, from which the current arsenic-rich groundwaters have evolved. Considering river basins within the above palaeo-hydrogeological framework provides a basis for assessing the risk of arsenic in groundwater basins of south and south-eastern Asia.     

Sequestration of cucurbitacin analogs by New and Old World chrysomelid leaf beetles in the tribe Luperini

Summary Two South American polyphagous leaf beetles,Diabrotica speciosa andCerotoma arcuata, selectively accumulated the bitter tasting compound 23,24-dihydrocucurbitacin D in their body after ingesting root tissues of cucurbit plants. Similarly, three Asian Cucurbitaceae-feeding specialists in the genusAulacophora were found to sequester the same compound. Cucurbitacin analogs were shown to deter feeding by a bird predator, indicating an allomonal role for these compounds in cucurbitacin-associated chrysomelid leaf beetles both of New and Old Worlds. The strong affinity to cucurbitacins, selective sequestration of the analogs and consequent protection from predators suggested an ecological adaptation mechanism developed in common among these two geographically isolated subtribes in the Luperini.     

Effects of Controlled Fire and Livestock Grazing on Bird Communities in East African Savannas

Abstract: In East Africa fire and grazing by wild and domestic ungulates maintain savannas, and pastoralists historically set fires and herded livestock through the use of temporary corrals called bomas. In recent decades traditional pastoral practices have declined, and this may be affecting biodiversity. We investigated the effects of prescribed fires and bomas on savanna bird communities in East Africa during the first and second dry seasons of the year (respectively before and after the rains that mark the onset of breeding for most birds). We compared abundance, richness, and community composition on 9‐ha burned plots, recently abandoned bomas, and control plots in the undisturbed matrix habitat over a 3‐year period. Generally, recently burned areas and abandoned bomas attracted greater densities of birds and had different community assemblages than the surrounding matrix. The effects of disturbances were influenced by interactions between primary productivity, represented by the normalized difference vegetation index, and time. Bird densities were highest and a greater proportion of species was observed on burned plots in the months following the fires. Drought conditions equalized bird densities across treatments within 1 year, and individuals from a greater proportion of species were more commonly observed on abandoned bomas. Yearly fluctuations in abundance were less pronounced on bomas than on burns, which indicate that although fire may benefit birds in the short term, bomas may have a more‐lasting positive effect and provide resources during droughts. Several Palearctic migrants were attracted to burned plots regardless of rainfall, which indicates continued fire suppression may threaten their already‐declining populations. Most notably, the paucity of birds observed on the controls suggests that the current structure of the matrix developed as a result of fire suppression. Traditional pastoralism appears critical to the maintenance of avian diversity in these savannas.     

Threats to intact tropical peatlands and opportunities for their conservation

Large, intact areas of tropical peatland are highly threatened at a global scale by the expansion of commercial agriculture and other forms of economic development. Conserving peatlands on a landscape scale, with their hydrology intact, is of international conservation importance to preserve their distinctive biodiversity and ecosystem services and maintain their resilience to future environmental change. We explored threats to and opportunities for conserving remaining intact tropical peatlands; thus, we excluded peatlands of Indonesia and Malaysia, where extensive deforestation, drainage, and conversion to plantations means conservation in this region can protect only small fragments of the original ecosystem. We focused on a case study, the Pastaza‐Marañón Foreland Basin (PMFB) in Peru, which is among the largest known intact tropical peatland landscapes in the world and is representative of peatland vulnerability. Maintenance of the hydrological conditions critical for carbon storage and ecosystem function of peatlands is, in the PMFB, primarily threatened by expansion of commercial agriculture linked to new transport infrastructure that is facilitating access to remote areas. There remain opportunities in the PMFB and elsewhere to develop alternative, more sustainable land‐use practices. Although some of the peatlands in the PMFB fall within existing legally protected areas, this protection does not include the most carbon‐dense (domed pole forest) areas. New carbon‐based conservation instruments (e.g., REDD+, Green Climate Fund), developing markets for sustainable peatland products, transferring land title to local communities, and expanding protected areas offer pathways to increased protection for intact tropical peatlands in Amazonia and elsewhere, such as those in New Guinea and Central Africa which remain, for the moment, broadly beyond the frontier of commercial development.     

Cost‐effective conservation of an endangered frog under uncertainty

How should managers choose among conservation options when resources are scarce and there is uncertainty regarding the effectiveness of actions? Well‐developed tools exist for prioritizing areas for one‐time and binary actions (e.g., protect vs. not protect), but methods for prioritizing incremental or ongoing actions (such as habitat creation and maintenance) remain uncommon. We devised an approach that combines metapopulation viability and cost‐effectiveness analyses to select among alternative conservation actions while accounting for uncertainty. In our study, cost‐effectiveness is the ratio between the benefit of an action and its economic cost, where benefit is the change in metapopulation viability. We applied the approach to the case of the endangered growling grass frog (Litoria raniformis), which is threatened by urban development. We extended a Bayesian model to predict metapopulation viability under 9 urbanization and management scenarios and incorporated the full probability distribution of possible outcomes for each scenario into the cost‐effectiveness analysis. This allowed us to discern between cost‐effective alternatives that were robust to uncertainty and those with a relatively high risk of failure. We found a relatively high risk of extinction following urbanization if the only action was reservation of core habitat; habitat creation actions performed better than enhancement actions; and cost‐effectiveness ranking changed depending on the consideration of uncertainty. Our results suggest that creation and maintenance of wetlands dedicated to L. raniformis is the only cost‐effective action likely to result in a sufficiently low risk of extinction. To our knowledge we are the first study to use Bayesian metapopulation viability analysis to explicitly incorporate parametric and demographic uncertainty into a cost‐effective evaluation of conservation actions. The approach offers guidance to decision makers aiming to achieve cost‐effective conservation under uncertainty.