A comparison of different lysis buffers to assess allele dropout from single cells for preimplantation genetic diagnosis

Single cell polymerase chain reaction (PCR) for preimplantation genetic diagnosis (PGD) requires high efficiency and accuracy. Allele dropout (ADO), the random amplification failure of one of the two parental alleles, remains the most significant problem in PCR-based PGD testing since it can result in serious misdiagnosis for compound heterozygous or autosomal dominant conditions. A number of different strategies (including the use of lysis buffers to break down the cell and make the DNA accessible) have been employed to combat ADO with varying degrees of success, yet there is still no consensus among PGD centres over which lysis buffer should be used (ESHRE PGD Consortium, 1999 ). To address this issue, PCR amplification of three genes (CFTR, LAMA3 and PKP1) at different chromosomal loci was investigated. Single lymphocytes from individuals heterozygous for mutations within each of the three genes were collected and lysed in either alkaline lysis buffer (ALB) or proteinase K/SDS lysis buffer (PK). PCR amplification efficiencies were comparable between alkaline lysis and proteinase K lysis for PCR products spanning each of the three mutated loci (ΔF508 in CFTR 90% vs 88%; R650X in LAMA3 82% vs 78%; and Y71X in PKP1 91% vs 87%). While there was no appreciable difference between ADO rates between the two lysis buffers for the LAMA3 PCR product (25% vs 26%), there were significant differences in ADO rates between ALB and PK for the CFTR PCR product (0% vs 23%) and the PKP1 PCR product (8% vs 56%). Based on these results, we are currently using ALB in preference to PK/SDS buffer for the lysis of cells in clinical PGD. Copyright © 2001 John Wiley & Sons, Ltd.     

Review of present European environmental policies and legislation involving cultural heritage

Cultural heritage is being addressed by a number of charters and conventions and it is clear that its consideration within decision-making processes is progressively becoming a real challenge, both for developers and public authorities. Against this background, this paper reviews the environmental assessment framework developed by the European Community, as this should increasingly influence decision making about cultural heritage in an urban setting. The legislative framework for access to environmental information is also reviewed because of its relevance to the decision-making process.The Directives on Environmental Impact Assessment and Strategic Environmental Assessment both require a consideration of cultural heritage in decision making. This requirement can go some way to addressing the paradox in the European Community's position whereby the community wishes to conserve and enhance its own cultural identity whilst, at the same time, cultural heritage is usually defined at a local level. The present paper suggests that, given their flexibility, the two European Community directives on environmental assessment constitute a promising opportunity to address this seeming paradox, but that there is a significant gap between legal obligations and the methodological tools to meet them.     

生态系统服务功能及其价值核算

生态系统服务功能是指生态系统服务与生态系统功能的综合，指生态系统与生态过程所形成及所维持的人类赖以生存的自然环境条件与效用，生态系统服务功能对于人类生存发展具有重要的价值，应该在社会经济体系中得以体现，本文探讨了生态系统服务功能的类型，价值核算方法与理论及其应用研究在完善国民经济核算体系、促进可持续发展等方面的重要意义，并对今后发展进行了展望。     

Evolution of environmental impact assessment as applied to watershed modification projects in Canada

This article reviews the application of environmental impact assessment (EIA) procedures and practices to three watershed modification projects situaled in western Canada. These ventures were justified for accelerating regional economic development, and cover the period during which public concerns for protecting the environment rapidly made their way into the national political agenda. An historical account and analysis of the situation, therefore, seems desirable in order to understand the development of EIA processes, practices, and methodologies since the start of construction of the first project in 1961. This study concludes that there has been good progress in predicting and evaluating environmental and related social impacts of watershed modification proposals. However, a number of obstacles need to be overcome before EIA can firmly establish itself as an effective planning tool. These difficulties include jurisdictional confusions and conflicts, division of authority and responsibility in designing and implementing appropriate mitigative and monitoring measures, lack of tested EIA methodologies, and limited availability of qualified human resources. A number of conclusions and suggestions are offered so that future watershed modification proposals may be planned and implemented in a more environmentally sustainable fashion. These include: (1) EIA processes must be completed before irrevocable decisions are made. (2) Any major intrusion into a watershed is likely to impact on some major components of the ecosystem(s). (3) Mitigation costs must form part of the benefit-cost analysis of any project proposal. (4) Interjurisdictional cooperation is imperative where watersheds cross political boundaries. (5) The EIA process is a public process, hence public concerns must be dealt with fairly. (6) The role of science in the EIA process must be at arms length from project proponents and regulators, and allowed to function in the interest of the protection of the environment and public health and safety. The views expressed here are the authors’ own and do not necessarily reflect those of FEARO and/or other government agencies and officials involved in the review of these projects.     

Fluctuations of Population Size and Structure in the Northern Mole Vole: Preliminary Analysis

Long-term stationary studies on the ecology of the northern mole vole (Ellobius talpinus Pall.), performed by the mark–recapture method from 1985 to 1997, have provided original data on population dynamics and structure. The analysis shows that, to reveal cyclic fluctuations of population size in this species, the period of three years should be taken as a unit of time for estimating the duration of one phase. The 12-year population cycle in E. talpinus has four distinct phases: an increase, a peak, a decline, and a minimum. At each phase, the population is characterized by certain features of family structure, age composition, birth and death rates, and the composition of migrants.     

Comparative Analysis of Ecophysiological Characteristics of Stephanodiscus hantzschii Grun. in the Periods of Its Bloom in Recreational Water Bodies

In the periods of summer and autumn bloom of the Stephanodiscus hantzschii Crun. in recreational water bodies, studies on the vertical distribution of chlorophyll a, its contents per unit biomass, efficiency in using photosynthetically active radiation (E_PhAR), and assimilative activity of microalgae were performed. The results confirmed the existence of two ecophysiological forms of St. hantzschii and provided evidence that both forms are typically autotrophic and can efficiently use low-intensity PhAR for photosynthesis.     

湖北省湿地的保护与利用

湿地是地球上具有多功能的、独特的生态系统，是人类最重要的生存环境之一。湖北省地处长江中游，湿地类型多，面积较大，河、湖湿地系统是主要的湿地类型。湖北湿地具有较大的资源潜力和环境调节功能，是自然界生物多样性丰富的生态景观。近年来，湖北省湿地保护面临人口增长的压力，围垦、污染和过度开发等对湿地保护构成了严重威胁，湿地环境遭到干扰和破坏。加强湿地科学的研究，保护管理好湖北湿地及其生态环境，实现湿地资源的持续利用，是湖北省2l世纪环保工作所面临的一项重要而紧迫的任务。     

北京市发展CNG汽车的条件与前景分析

北京市汽车排放污染物贡献率已达到：HC73.5％，CO63．4％，NOx46％。用CNG代替汽油作汽车燃料，可使CO减少97％、HC减少72％、NOx减少39％、CO2减少24％、SO2减少90％。建立CNG加气站和CNG汽车改装这两个CNG应用关键问题在技术上、经济上都是可行的。北京市CNG燃料来源有保障。北京市车辆都比较适合使用CNG气体燃料。北京有很强的CNG汽车产业科技力量。对北京市当前发展CNG汽车产业提出了一些建议。     

论环境产业

本文在可持续发展视角下,把环境的保护、修复、维护的生产活动及与之直接相关的服务活动称为环境产韭,包括发挥减少、避免污染环境作用的资源再利用鼗和发挥治理污染物与修复、改善环境作用的环境维护业.通过探讨环境产业的内涵、发展历程、运行机制和在产业体系中的位置与作用,指出环境产业是在法规保障下政府强行推动产生和发展起来的,是产业体系中不可缺少的重要组成部分;考虑自然环境对社会行为的影响,改进政府行为构建适台环境产业与传统产业协调发展的经济体制是加快发展环境产业的关键.     

Biological Activity in a Heavily Organohalogen-Contaminated River Sediment (8 pp)

Background, Aims and Scope Sediments of the Spittelwasser creek are highly polluted with organic compounds and heavy metals due to the discharge of untreated waste waters from the industrial region of Bitterfeld-Wolfen, Germany over the course of more than one century. However, relatively few data have been published about the chloroorganic contamination of the sediment. This paper reports on the content of different (chloro)organic compounds with special emphasis on polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F), and chlorobenzenes. Existing concepts for the remediation of Spittelwasser sediment include the investigation of natural attenuation processes, which largely depend on the presence of an intact microbial food web. In order to gain more insight in terms of biological activity, we analyzed the capacity of sediment microflora to degrade organic matter by measuring the activities of extracellular hydrolytic enzymes involved in the biogeochemical cycling of carbon, nitrogen, phosphorus and sulfur. Furthermore, the detection of physiologically active bacteria in the sediment, particularly of those known for their capability to reductively dehalogenate organochlorine compounds, illustrates the potential for intrinsic bioremediation processes. Methods PCDD/F and chlorobenzenes were analyzed by gas chromatography(GC)/mass spectrometry and GC/flame ionization detection, respectively. The activities of hydrolytic enzymes were determined from freshly sampled sediment layers using 4-methylumbelliferyl (MUF) or 7-amino-4-methylcoumarin-conjugated model compounds and kinetic fluorescence measurements. Physiologically active bacteria from different sediment layers were microscopically visualized by fluorescence in situ hybridization (FISH). Specific bacteria were identified by 16S rRNA gene amplification and sequencing. Results and Discussion The PCDD/F congener profile was dominated by dibenzofurans. In addition, the presence of specific tetra and pentachlorinated dibenzofurans supported the assumption that extensive magnesium production was one possible source for the high contamination. A range of other chloroorganic compounds, including several isomers of chlorobenzenes, hexachlorocyclohexane and 1,1,1-trichloro-2,2-bis (p-chloro-phenyl)ethane (DDT), was present in the sediment. Activities of extracellular hydrolytic enzymes showed a strong decrease in those sediment layers that were characterized by high contents of absorbable organic halogen (AOX), indicating disturbed organic matter decay. Interestingly, an abnormal increase of cellulolytic enzyme activities below the organochlorine-rich layers was observed, possibly caused by residual cellulose from discharges of sulfite pulping wastes. FISH revealed physiologically active bacteria in most sediment layers from the surface down to the depth of about 60 cm, including members of Desulfitobacterium (D.) and Sulfurospirillum. The presence of D. dehalogenans was confirmed by its partial 16S rRNA gene sequence. Conclusions Results of chemical sediment analyses demonstrated high loads of organochlorine compounds, particularly of PCDD/F. Several years after stopping the waste water discharge to Spittelwasser creek, this sediment remains a main source for pollution of the downstream river system by way of the ongoing mobilization of sediment during high floods. As indicated by our enzyme activity measurements, the decomposition potential for organic matter is low in organochlorine-rich sediment layers. In contrast, the comparably higher enzyme activities in less organochlorine-polluted sediment layers as well as the presence of physiologically active bacteria suggest a considerable potential for natural attenuation. Recommendations and Perspectives From our data we strongly recommend to explore the degradative capacity of sediment microorganisms and the limits for in situ activity towards specific sediment pollutants in more detail. This will give a sound basis for the integration of bioremediation approaches into general concepts to reduce the risk that permanently radiates from this highly contaminated sediment. Submission Editor: Dr. Henner Hollert (Henner.Hollert@urz.uniheidelberg.de)