Preimplantation genetic diagnosis of the fragile X syndrome by use of linked polymorphic markers

Fragile X syndrome is the most common cause of familial mental retardation. The most common mutation is expansion of a triplet (CGG)_n repeat in the 5′ untranslated region of the FMR1 gene on Xq27.3. The expansion is refractory to PCR due to preferential amplification of the smaller allele in heterozygous cells and the high GC content of the repeat and surrounding sequences. Direct detection of the normal parental alleles in preimplantation embryos has been used for preimplantation genetic diagnosis (PGD) of this disorder. However, this approach is only suitable for approximately 63% of couples due to the heterozygosity of the repeat in the normal population. As an alternative we investigated the use of polymorphic markers flanking the mutation to track the normal and premutation carrying maternal chromosomes in preimplantation embryos. Using a panel of 11 polymorphisms, six (CA)_n repeats and five single nucleotide polymorphisms, diagnosis was developed for 90% of referred couples. Multiplex amplification of informative markers was tested in 300 single buccal cells from interested couples with efficiency and allele drop out (ADO) rates ranging from 69% to 96% and 6% to 18%, respectively. Use of this approach is accurate and applicable to a larger number of patients at risk of transmitting fragile X to their offspring. Copyright © 2001 John Wiley & Sons, Ltd.     

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.     

铁锰复合氧化物滤料去除地下水中四环素的效能

以化学挂膜成熟运行5a的铁锰复合氧化物滤料为研究对象,利用静态与连续流柱实验相结合的方式,探究了该滤料去除四环素的效果与机理.结果表明,去除过程符合准二级反应动力学模型;且滤料质量越大,初始浓度越高,对四环素去除的效果越好;当水中共存阳离子浓度为0.01mol/L时,Mg²⁺抑制四环素的去除,而Ca²⁺、K⁺对四环素的去除过程无显著影响.连续流柱实验与发光细菌毒性检测分析,表明,铁锰氧化物滤料对四环素去除效果显著高于石英砂,且铁锰复合氧化物滤料去除四环素后急性毒性降低.此外,机理探究实验结果证实了铁锰复合氧化物滤料去除四环素是吸附与氧化的过程.     

焚烧飞灰重金属含量及浸出长期变化规律研究

通过对24个飞灰样品的重金属全量消解和高精度X射线荧光光谱法（HDXRF）快速检测结果进行线性回归分析,发现HDXRF法可较准确测定飞灰中Cu、Pb、Zn、Ni和Cd含量（线性回归决定系数R²³0.90）.采用HDXRF法对某焚烧厂飞灰重金属含量6个月快速检测及浸出毒性测定数据统计分析发现：飞灰重金属月均含量较稳定（变异系数CV<0.14）但日均含量波动较大（CV>2.54）;飞灰中Pb、Cu和Zn含量呈正相关（相关系数r>0.78）;金属浸出浓度受浸出液终点pH值影响显著,超标频率依次为Pb（68.9%）、Cd（66.1%）、Ni（24.0%）、Cu（14.2%）和Zn（1.6%）.短期飞灰金属含量和浸出毒性数据受焚烧工况,烟气处理工况和垃圾组分等多种因素共同影响,不具有代表性.为评估垃圾分类源头削减重金属效果,建议采用快速检测方法对焚烧飞灰重金属含量进行长期监测.     

内质网应激在氟暴露致小鼠睾丸损伤中的作用

探讨了内质网应激在亚慢性氟暴露致小鼠睾丸损伤中的作用及分子机制.选用健康初断乳ICR雄性小鼠30只,随机分为对照组（C）、低氟组（LF）和高氟组（HF）,分别饮用自来水、5、30 mg·L^-1氟化钠水溶液90 d.亚慢性氟暴露结束后,以睾丸脏器系数、睾丸组织氧化/抗氧化酶和形态结构、精子质量、睾丸细胞凋亡、葡萄糖调节蛋白78（GRP78）、CCAAT/增强子结合蛋白同源蛋白（CHOP）、半胱氨酸天冬氨酸蛋白酶12（CASPASE-12）、半胱氨酸天冬氨酸蛋白酶3（CASPASE-3）为观测点.结果表明,与对照组比,LF组和HF组LDH、SOD、T-AOC活性下降,MDA含量上升,HF组GSH-PX活性下降,差异有统计学意义（p<0.05或p<0.01）;LF组可见细胞层次减少、间隙变大,成熟精子数量减少,HF组细胞溶解、层次紊乱,空泡化严重,少见成熟精子;LF组和HF组小鼠的精子活力降低,HF组小鼠精子数量下降,畸形率上升,差异有统计学意义（p<0.05或p<0.01）;LF组和HF组睾丸细胞凋亡指数上升,差异有统计学意义（p<0.01）;LF组和HF组Grp78、Caspase-12、Caspase-3基因表达水平上升,差异有统计学意义（p<0.05或p<0.01）.结果提示,除氧化应激以外,Caspase-12和Caspase-3基因表达异常可能是氟暴露致小鼠睾丸细胞凋亡异常的分子机制之一.     

石墨烯/聚苯胺修饰阴极对反硝化MFC性能影响

反硝化生物阴极微生物燃料电池（MFC）以电极为电子供体,在自养条件下完成硝酸盐去除过程.本研究以碳布（CC）为基底材料,分别制备获得还原氧化石墨烯修饰（rGO-CC）,聚苯胺修饰（PANI-CC）及二者复合修饰的CC电极（rGO/PANI-CC）,并考察其作为阴极材料对反硝化生物阴极MFC产电脱氮性能的影响.扫描电镜结果显示,rGO-CC和PANI-CC的碳纤维分别被片层状rGO和网状PANI覆盖,而rGO/PANI-CC表面呈现PANI在附着rGO的碳纤维上团聚的形貌,均增大了碳布的比表面积.循环伏安测试显示,rGO/PANI-CC具有最高的电化学活性.以rGO-CC,PANI-CC和rGO/PANI-CC为阴极构建MFC的产电能力分别提高了82%,24%和41%,其阴极对NO₃^--N的去除能力增强了23%,9%和13%.16S rDNA测序结果揭示修饰后电极表面微生物的多样性下降,Stappia和Pacacoccus属微生物的丰度增加.     

Performance of different macrophytes in the decontamination of and electricity generation from swine wastewater via an integrated constructed wetland-microbial fuel cell process

Plants constitute a major element of constructed wetlands(CWs).In this study,a coupled system comprising an integrated vertical flow CW(IVCW) and a microbial fuel cell(MFC) for swine wastewater tre atment was developed to research the effects of macrophytes commonly employed in CWs,Canna indica,Acorus calamus,and Ipomoea aquatica,on decontamination and electricity production in the system.Because of the different root types and amounts of oxygen released by the roots,the rates of chemical oxygen demand(COD) and ammonium nitrogen(NH_4~+-N) removal from the swine wastewater differed as well.In the unplanted,Canna indica,Acorus calamus,and Ipomoea aquatica systems,the COD removal rates were 80.20%,88.07%,84.70%,and 82.20%,respectively,and the NH_4~+-N removal rates were 49.96%,75.02%,70.25%,and 68.47%,respectively.The decontamination capability of the Canna indica system was better than those of the other systems.The average output voltages were 520±42,715±20,660±27,and 752±26 mV for the unplanted,Canna indica,Acorus calamus,and Ipomoea aquatica systems,respectively,and the maximum power densities were 0.2230,0.4136,0.3614,and0.4964 W/m~3,respectively.Ipomoea aquatica had the largest effect on bioelectricity generation promotion.In addition,electrochemically active bacteria,Geobacter and Desulfuromonas,were detected in the anodic biofilm by high-throughput sequencing analysis,and Comamonas(Proteobacteria),which is widely found in MFCs,was also detected in the anodic biofilm.These results confirmed the important role of plants in IVCW-MFCs.     

Identification of potential electrotrophic microbial community in paddy soils by enrichment of microbial electrolysis cell biocathodes

Electrotrophs are microbes that can receive electrons directly from cathode in a microbial electrolysis cell (MEC). They not only participate in organic biosynthesis, but also be crucial in cathode-based bioremediation. However, little is known about the electrotrophic community in paddy soils. Here, the putative electrotrophs were enriched by cathodes of MECs constructed from five paddy soils with various properties using bicarbonate as an electron acceptor, and identified by 16S rRNA-gene based Illumina sequencing. The electrons were gradually consumed on the cathodes, and 25%–45% of which were recovered to reduce bicarbonate to acetic acid during MEC operation. Firmicutes was the dominant bacterial phylum on the cathodes, and Bacillus genus within this phylum was greatly enriched and was the most abundant population among the detected putative electrotrophs for almost all soils. Furthermore, several other members of Firmicutes and Proteobacteria may also participate in electrotrophic process in different soils. Soil pH, amorphous iron and electrical conductivity significantly influenced the putative electrotrophic bacterial community, which explained about 33.5% of the community structural variation. This study provides a basis for understanding the microbial diversity of putative electrotrophs in paddy soils, and highlights the importance of soil properties in shaping the community of putative electrotrophs.     

A ten year summary of concurrent ambient water column and sediment toxicity tests in the Chesapeake Bay watershed: 1990-1999

The goal of this study was to identify the relative toxicity ofambient areas in the Chesapeake Bay watershed by using a suiteof concurrent water column and sediment toxicity tests at seventy-five ambient stations in 20 Chesapeake Bay rivers from1990 through 1999. Spatial and temporal variability was examinedat selected locations throughout the 10 yr study. Inorganicand organic contaminants were evaluated in ambient water andsediment concurrently with water column and sediment tests toassess possible causes of toxicity although absolute causalitycan not be established. Multivariate statistical analysis wasused to develop a multiple endpoint toxicity index (TOX-INDEX) at each station for both water column and sediment toxicity data. Water column tests from the 10 yr testing period showed that49% of the time, some degree of toxicity was reported. The mosttoxic sites based on water column results were located inurbanized areas such as the Anacostia River, Elizabeth River andthe Middle River. Water quality criteria for copper, lead,mercury, nickel and zinc were exceeded at one or more of thesesites. Water column toxicity was also reported in localizedareas of the South and Chester Rivers. Both spatial and temporalvariability was reported from the suite of water column toxicitytests. Some degree of sediment toxicity was reported from 62% of the tests conducted during the ten year period. The ElizabethRiver and Baltimore Harbor stations were reported as the most toxic areas based on sediment results.Sediment toxicity guidelines were exceeded for one or more of thefollowing metals at these two locations: arsenic, cadmium,chromium, copper, lead, nickel and zinc. At the Elizabeth Riverstations nine of sixteen semi-volatile organics and two of sevenpesticides measured exceeded the ER-M values in 1990. Ambientsediment toxicity tests in the Elizabeth River in 1996 showedreduced toxicity. Various semi-volatile organics exceeded the ER-M values at a number of Baltimore Harbor sites; pyrene anddibenzo(a,h)anthracene were particularly high at one of thestations (Northwest Harbor). Localized sediment toxicity was alsoreported in the Chester, James, Magothy, Rappahannock, andPotomac Rivers but the link with contaminants was not determined.Both spatial and temporal variability was less for sedimenttoxicity data when compared with water column toxicity data. Acomparison of water column and sediment toxicity data for thevarious stations over the 10 yr study showed that approximatelyhalf the time agreement occurred (either both suite of testsshowed toxicity or neither suite of tests showed toxicity).     

Response of the Ascorbate-Peroxidase of Selenastrum capricornutum to Copper and Lead in Stormwaters

The green alga Selenastrum capricornutum expresses a uniqueascorbate peroxidase, that responds to copper and lead. Attemptswere made to test if this peroxidase could be used to monitor thelevels of copper and lead in natural waters. When S.capricornutum was exposed to a stormwater sample, the specificactivity of the peroxidase in the cell extract was commensuratewith the combined copper and lead contents in the sample. Theperoxidase responses were also correlated with the 96 hr biomasstoxicity assay of S. capricornutum. However, unlike thebiomass toxicity assay, the peroxidase activity was not affectedby the anions in the samples. The use of this peroxidase can beused as a marker for testing heavy metal toxicity in the water.