Prenatal diagnosis of Apert syndrome: report of two cases

Two de novo cases with Apert Syndrome detected prenatally are presented herein. In the first, fetal ultrasound findings of syndactyly of the hands, craniosynostosis and proptosis resulted in a prenatal diagnosis in the nineteenth week of gestation. This is the earliest prenatal diagnosis of this syndrome in a not-at-risk case. Following counseling, this pregnancy was terminated and subsequent pathological examination and DNA analysis confirmed the diagnosis of Apert Syndrome and coarctation of the aorta. In the second case, fetal ultrasound at 21 weeks' gestation revealed a hypoplastic left heart and clover-leaf skull. Following counseling, this pregnancy was also terminated. Further examination of the fetus and DNA analysis led to a diagnosis of Apert Syndrome. These cases emphasize the need to complete a thorough fetal ultrasound in cases with potentially lethal cardiac abnormality and the importance of incorporating a fetal pathologist, as well as a medical geneticist, in the investigations performed after delivery or pregnancy termination when a fetal abnormality is detected on ultrasound. Copyright © 2003 John Wiley & Sons, Ltd.     

Exploring Chemical Variables in Ligustrum lucidum Ait. F. Tricolor (Rehd.) Rehd. in Relation to Air Pollutants and Environmental Conditions

Ligustrum lucidum Ait. f. tricolor (Rehd.) Rehd. in relation to atmospheric pollutants in Córdoba city, Argentina. The study area receives regional pollutants and was categorized taking into account traffic level, industrial density, type of industry, location of the sample point in relation to the street corner, treeless condition, and topographic level. Dried weight/fresh weight ratio (DW/FW) and specific leaf area (SLA) were calculated, and concentrations of chlorophylls, carotenoids, total sulfur, soluble proteins, malondialdehyde (MDA), and hydroperoxy conjugated dienes (HPCD) were determined in leaf samples. Sulfur content correlates positively with traffic density and SLA correlates negatively with some combinations of the categorical variables; MDA correlates positively with topographic level and total protein concentration correlates negatively with treeless condition. On the basis of our results, traffic, location of trees, type of industry, situation of a tree with respect to others, and topographic level are the environmental variables to bear in mind when selecting analogous sampling points in a passive monitoring program. An approximation to predict tree injury may be obtained by measuring DW/FW ratio, proteins, pigments, HPCD, and MDA as they are responsible for the major variability of data.     

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.     

生态足迹影响因子的定量分析

人口的膨胀和人类工业化进程的加剧，使得人类向自然界获取越来越多的资源，同时向环境源源不断的输入废弃物，已经严重超过了自然生态系统的供给能力和环境容量，生态环境日益恶化，水土流失、草场退化、植被消亡、生物多样性锐减、全球变暖等情况已经严重到难以遏制的地步，危及到人类自身的生存。在和平与发展成为世界两大主题的今天，人们越来越关注可持续发展的问题。在现有资料的基础上，利用主成分分析方法，定量地讨论了中国各省（区市）1999年生态足迹大小与其影响因子间的关系。结果表明，生态足迹的大小是各省（区市）的大中型企业个数、全社会固定生产投资等众多因子共同作用的结果，其中总人口和GDP是生态足迹大小的主要影响因子，其因子载荷量分别达到了0.940和0.913。值得一提的是非农业人口与生态足迹的相关系数超过了农业人口，这说明由于消费模式和生活水平等的差异，非农业人口对生态足迹的影响大于农业人口对生态足迹的影响。在此基础上建立了生态足迹影响因子的多元线性回归模型，以期为生态足迹在进行区域可持续发展评价的方法上提供新的思路。     

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。     

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.     

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。     

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.     

Pitfalls in molecular diagnosis in a family with severe factor VII (FVII) deficiency—misdiagnosis by direct sequence analysis using a PCR product

Molecular diagnostic tests are becoming a routine analysis in many laboratories. These modern analyses are widely used in clinical medicine, forensic, genetic and prenatal diagnosis and also in preimplantation genetic diagnosis. The accuracy of analysis is highly dependent on the success achieved in minimising genotyping errors. The pitfalls in molecular diagnostic tests can be due to a simple technique such as the polymerase chain reaction (PCR) used universally. This technique is routinely used for its apparent accuracy, but it is also a well-known source of errors. We report an error introduced during PCR reaction that leads to a wrong sequence result and consequently to a ‘false’ molecular result in a next prenatal diagnosis in a family with severe factor VII (FVII) deficiency. This error was verified using an unsuitable primer design in a rich repetitive sequence of the FVII gene that leads to a false annealing and then to a wrong molecular diagnosis. It is essential to link closely molecular data with clinical and phenotype analysis in order to avoid false-negative or false-positive results, which is of great importance to diagnosis and molecular prevention. Copyright © 2003 John Wiley & Sons, Ltd.