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.     

中国可持续社会养老保险的综合评价体系和实证分析

我国可持续社会养老保险的综合评价指标体系包括四个层次和三级子系统，内容涉及养老保险人数、机构设置、基金管理和代际平衡四个子目标。应用因子分析模糊综合评价方法对我国1990-2001年养老保险建设状况进行评价，本文认为基金管理，行政管理和代际失衡是影响我国可持续养老保险发展的主要因素。     

可持续旅游及其区域实现途径探讨—以井冈山风景名胜区为例

可持续旅游是 80年代中期以后提出的关于旅游业发展的新思想。经过十多年的不断拓展深化 ,已经得到世界各国的广泛认同 ,并作为跨世纪旅游业发展的战略指导思想。本文从可持续旅游的由来和对可持续旅游内涵实质的界定角度出发 ,结合可持续旅游实现途径的探讨 ,以井冈山风景名胜区为例 ,对风景区旅游业可持续发展进行了深入的探讨 ,为景区的发展规划提出了战略性构想 ,以期能对井冈山风景名胜区旅游资源的可持续利用和旅游业的可持续发展起到抛砖引玉的作用     

南宁城市大气污染对人体健康的危害及治理对策

南宁市大气属煤烟型污染 ,大气的主要污染物为SO2 、NOx、TSP、降尘 ,虽然随着环境管理和污染治理工作的加强 ,污染物浓度逐年下降 ,但是工业区大气污染仍然较重。污染物流行病学调查显示 :工业区癌症和呼吸系统疾病死亡率均高于全市平均水平 2倍左右 ,城区又高于郊区 2倍 ,大气污染综合指数与呼吸内科门诊就诊人数呈正相关。用邓聚龙的灰色系统理论分析得知污染物对癌症和呼吸系统疾病死亡率有关联 ,关联度从大到小排序为 :TSP >降尘 >SO2 >NOx ,最后提出了大气污染治理的对策。     

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.     

新时期区域公共管理创新

信息社会发展与全球化趋势，出现了区域集群、区域竞争、区域创新、学习区域等新特征，给区域管理者带来了新的挑战。区域发展需要创新，区域公共管理同样需要创新。本文从新公共管理、区域创新系统、区域核心竞争力、支持系统等方面对区域管理创新进行了探讨。     

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。     

现行财税体制下我国耕地资源数量变化的实证研究

耕地资源数量变化受到人口、经济等诸多因素的影响，而财税体制是影响耕地资源数量变化的更深屡次原因。通过数量关系的初步观察和作用机理分析说明我国现行财税体制对耕地资源数量变化存在正反两方面作用，总体来看，对耕地资源保护不利。计量结果进一步证实了我国现行财税体制对耕地资源数量变化存在显著影响。揭示了其影响的方向和大小。根据研究结果，文章有针对性地提出了增加地方财政预算收入，合理分配土地出让金比例和科学使用土地出让金等相关政策建议。     

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)