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.     

黄河流域水资源价值的计算与分析

利用经济杠杆的调节作用促使水资源的高效利用，是缓和水资源供需矛盾的有效措施之一。本文以水资源价值理论为基础，从黄河全流域水资源优化配置角度出发，以利用水资源获得最大经济效益为目标，测算了黄河水资源的理论价值，为制定黄河水资源费的征收标准提供科学的理论依据。     

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.     

母岩的风化剥蚀速率与土壤允许流失量的关系 --以长江三峡坝区风化花岗岩土壤为例

土壤允许流失量的确定是一个非常复杂而又必须解决的问题，它关系到水土保持措施的布设和土壤的可持续利用。然而，发育在不同母质上的土壤，其土壤的最大允许流失量差异很大，确定这一值的依据也各不相同。在岩成土壤地区，母岩风化剥蚀速率的大小直接影响土壤的发育。是确定土壤允许流失量、分析人类加速水土流失的重要依据之一。选长江三峡黄陵背斜段风化花岗岩土壤为研究对象，根据剥蚀沉积相关原理，通过恢复古地理环境及时代，计算出新生代以来本区花岗岩的平均风化剥蚀速率为16．97mm／ka．最大剥蚀速率为49．56。又根据区内太平溪流域的泥沙资料，算出了当地现代的剥蚀速率，多年平均为297．7mm／ka，最小值为31．5mm／ka，而水利部颁布的当地土壤允许流失量为200t／km^2,a，折合为76．9mm／ka，二者相差近1．5—4．5倍。基于此。提出了确定土壤允许流失量必须参考母岩风化剥蚀速率的新观点。     

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。     

Industrial Cluster Upgradation of Regenerated Resources in Miluo City

With China's rapid economic growth, ecological construction and environmental protection become increasingly important. The regenerated resources industry is an effective way to solve problems, such as resources depletion, energy shortage, and pollution, and it also has strategic importance for the construction of a resource-conserving and environment-friendly society. The regenerated resources industry has been established in Miluo for long time, which includes a recycling system, a processing and utilization system, and a refuse decontamination system. An industrial cluster is in its early stage of development. In order to solve current problems, such as short industrial chain, low processing rate, and low added value, the industrial cluster should be dynamically upgraded by means of technology innovation, chain nucleus creation, and chain extension. We think the industrial cluster of regenerated resources will become a local brand for Miluo, from which other regions or cities will gain valuable experiences and inspirations.     

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)     

SUSTAINABILITY AT THE LOCAL LEVEL: MANAGEMENT TOOLS AND MUNICIPAL TAX INCENTIVE MODEL

The implementation of sustainable development may seem a simple concept when written on paper. However to carry-out long term actions put forward by the Agenda 21 (AG21) at the local level represents one of the main challenges as municipal governments in general do not have the capacity to effectively implement the process. Regional environmental assessment (REA) has shown to be effective in supporting decision-making not only to correct environmental problems due to past unsustainable social-economic developments but also help local governments to implement sustainable actions. However this requires long-term investments of AG21 plans and projects. The allocation of regular and consistent financial resources is one of the main ingredients for the sustainable development process. But traditional plans and projects financed by national and/or international funds may not be sustainable in the long-term because they become dependent on external funding. Research demonstrate that innovative economic instruments such as ecotaxes represent a feasible alternative to sponsor local sustainability because taxes are collected permanently by the government and could be invested in continuous actions. Ecotaxes experiences have provided important reference to structure a municipal incentive model (MIM) to sponsor AG21’s environmental plans and projects on a long-term (permanent) basis. However sustainable development cannot be solely through economic investments. A comprehensive municipal environmental management scheme (MEMS) has been established to support the incentive model. The scheme seeks not only to improve local institutional framework but also incentive continuous participation of local stakeholders at all levels of society. Participatory events and the provision of incentives (educational and financial) are key to motivate society to protect the environment and support actively the sustainable development process as emphasised in the RIO-92 Conference.