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.     

Male-biassed response of garden chafer, Phyllopertha horticola L., to leaf alcohol and attraction of both sexes to floral plant volatiles

Summary. Field tests were performed to evaluate the response of the garden chafer, Phyllopertha horticola (Coleoptera: Scarabaeidae) to known scarab attractants and to investigate the influence of trap colour and height on the number of captured beetles. Both sexes were attracted by a lure mixture composed of geraniol, eugenol, and 2-phenylethyl propionate (PEP) (ratio 3:7:3). When testing floral volatiles individually, only geraniol, eugenol and methyl anthranilate but not PEP was attractive. Response of garden chafer to (Z)- 3-hexen-1-ol was strongly male-biassed. Both the response to floral volatiles and (Z)-3-hexen-1-ol was increased by using funnel traps with yellow instead of grey vanes. Traps positioned at 50 and 125 cm above ground captured significantly more garden chafers than those at 200 cm. The strongly male-biassed response to (Z)-3-hexen-1-ol suggests that in P. horticola (subfamily Rutelinae) orientation towards plant volatiles emitted upon mechanical damage of plants is part of the male mate finding strategy as recently demonstrated for cockchafers of the genus Melolontha (subfamily Melolonthinae). Possible application of plant volatiles for control of P. horticola is discussed.     

广州轨道交通三号线高速行驶条件下对安全限界的影响研究

给出了轨道交通车辆横向偏移量的计算方法 ,并应用列车 -线路动力耦合模型 ,分析了广州地铁三号线的安全限界变化问题。确定了当前按 80km/h行车速度设计的建筑限界能满足 13 0km/h行车速度的要求 ,限界尺寸不需改动可保证列车运行安全平稳     

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.     

长江三峡区间数字流域水系的构建

基于全球陆地一公里基础高程GLOBE数据 ,采用Martz和Garbrecht研制的数字高程流域水系模型自动提取长江三峡万县～宜昌区间的河网水系、各子流域分水线、河网与子流域编码及河网结构拓扑关系 ,从而构成该区域的数字流域和数字水系。结果表明该模型生成的水系是可以接受的 ,与 1:10万地形图上长江三峡万县～宜昌区间的水系一致 ,这为空间分布式模型的建立提供了必备的空间信息数据。最后 ,探讨了数字流域水系在流域生态学中的应用前景 ,构建的数字水系可为中尺度以上流域水体和水生生物的资源保护与合理利用提供决策平台 ,服务于社会经济的可持续发展。     

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。     

湖泊水质模型研究进展

湖泊是最重要的淡水资源之一，对社会和经济的发展起着不可估量的作用。保护和改善湖泊水环境问题已成为当前世界关注的一个焦点。湖泊水质模型是一种利用数学语言来描述湖泊污染过程中的物理、化学、生物化学及生物生态各方面之内在规律和相互联系的手段。作为湖泊水环境污染治理、规划决策分析中的一个重要工具，它可以为湖泊的综合整治和科学管理提供科学的依据，在环境保护领域中发挥着举足轻重的作用。分别介绍了湖泊水质模型国内外研究动态、类型、常用软件（WASP、EFDC、CE QUAL W2、CE QUAL R1、CE QUAL ICM、MIKE、SMS等）和应用实例，并综观湖泊水质模型的研究历史和应用前景，系统分析了湖泊水质模型研究的发展趋势。     

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)     

草原旅游发展中旅游管理部门环境意识的调查研究

根据环境意识结构．设计了旅游管理部门环境意识调查问卷。其主要的指标体系包括：旅游环境知识水平、旅游环境态度、旅游环境评价和旅游环境行为四个方面。对指标体系各部分之间以及每一指标部分的每一题目之间的相对重要性进行权重的确定．而且对各部分的每道题目的各个选项予以赋分。基于权重和赋分．结合调查问卷的实际情况，设计了指标体系各部分评价模型和环境意识总体评价模型。以此来计算草原旅游发展中旅游管理部门环境意识水平的综合得分。经过分析得知，旅游管理部门环境意识模式是“环境知识制约型”模式。