First-trimester fetal sex determination in maternal serum using real-time PCR

An Erratum has been published for this article in Prenatal Diagnosis 22(13) 2002, 1241. Fetal sex prediction can be achieved using PCR targeted at the SRY gene by analysing cell-free fetal DNA in maternal serum. Unfortunately, the results reported to date show a lack of sensitivity, especially during the first trimester of pregnancy. Therefore, determination of fetal sex by maternal serum analysis could not replace karyotype analysis following chorionic villus sampling. A new highly sensitive real-time PCR was developped to detect an SRY gene sequence in maternal serum. Analysis was performed on 121 pregnant women during the first trimester of pregnancy (mean gestational age: 11.8 weeks). Among them, 51 had at least one previous male-bearing pregnancy. Results were compared with fetal sex. SRY PCR analysis of maternal serum was in complete concordance with fetal sex. Among the 121 pregnant women, 61 were bearing a male fetus and 60 a female fetus. No false-negative results were observed. Furthermore, no false-positive results occurred, even though 27 women carrying a female fetus during the current pregnancy had at least one previous male-bearing pregnancy. This study demonstrates that a reliable, non-invasive sex determination can be achieved by PCR analysis of maternal serum during the first trimester of pregnancy. This non-invasive approach for fetal sex prediction should have great implications in the management of pregnant women who are carriers of an X-linked genetic disorder. Prenatal diagnosis might thus be performed for male fetuses only, avoiding invasive procedures and the risk of the loss of female fetuses. Copyright © 2001 John Wiley & Sons, Ltd.     

Trophoblast-like cells sorted from peripheral maternal blood using flow cytometry: A multiparametric study involving transmission electron microscopy and fetal dna amplification

Three monoclonal antibodies (MAbs) against trophoblast (GB17, GB21, and GB25) and flow cytometry were used to sort trophoblast-like cells (TLCs) from peripheral blood of pregnant women. Sorted TLCs were processed for electron microscopy and fetal DNA amplification of the Y-specific sequences from mothers carrying male fetuses. At the ultra-structural level, most of the nucleated cells had the morphology of leucocytes, suggesting maternal contaminants, and we did not find the characteristic features of the free inter-villous trophoblast cells. Nevertheless, polymerase chain reaction (PCR) analysis showed an amplification of Y-specific sequences in two out of three samples of sorted TLCs. These results suggest that besides the maternal leucocytes, sufficient trophoblast nucleated fetal cells can be obtained using cell enrichment by sorting. This sensitive method holds promise for non-invasive prenatal diagnosis of fetal sex and if sufficient Y(positive) nuclei are found, for the diagnosis of selected numerical chromosome abnormalities.     

Hunting Increases Dispersal Limitation in the Tree Carapa procera, a Nontimber Forest Product

Abstract:  The sustainability of seed extraction from natural populations has been questioned recently. Increased recruitment failure under intense seed harvesting suggests that seed extraction intensifies source limitation. Nevertheless, areas where more seeds are collected tend to also have more intense hunting of seed-dispersing animals. We studied whether such hunting, by limiting disperser activity, could cause quantitative dispersal limitation, especially for large crops and for crops in years of high seed abundance. In each of four Carapa procera (Meliaceae) populations in French Guiana and Surinam, two with hunting and two without, we compared seed fate for individual trees varying in crop size in years of high and low population-level seed abundance. Carapa seeds are a nontimber forest product and depend on dispersal by scatter-hoarding rodents for survival and seedling establishment. Hunting negatively affected the proportion of seeds dispersed and caused greater numbers of seeds to germinate or be infested by moths below parent trees, where they would likely die. Hunting of seed-dispersing animals disproportionally affected large seed crops, but we found no additional effect of population-level seed abundance on dispersal rates. Consistently lower rates of seed dispersal, especially for large seed crops, may translate to lower levels of seedling recruitment under hunting. Our results therefore suggest that the subsistence hunting that usually accompanies seed collection is at the cost of seed dispersal and may contribute to recruitment failure of these nontimber forest products. Seed extraction from natural populations may affect seedling recruitment less if accompanied by measures adequately incorporating and protecting seed dispersers.     

长江中下游环境DNA宏条形码生物多样性检测技术初步研究

长江生物多样性在人为影响下面临严重威胁,物种监测是生物多样性保护的基础,为完善长江水生态监测体系,实现高效无损伤的物种监测,在长江中下游干流3个江段（新滩、安庆和芜湖）采集水样,建立长江水样环境DNA宏条形码物种检测体系并评估其有效性.结果表明:①长江中下游环境DNA宏条形码检测到32个物种,包括20种鱼类、1种水生哺乳动物（长江江豚）和11种陆生动物,其中鱼类物种包括鲤形目、鲇形目、鲈形目和鲱形目,其种数占鱼类总种数的比例分别为60%、25%、10%和5%.②长江中下游渔获物中资源量居首位的鲤形目在环境DNA调查中序列数最多,占鱼类总序列的96.2%,其次为鲱形目（占比为3.5%）,鲇形目和鲈形目占比较低,分别为0.2%和0.1%,4个类目序列相对丰度与渔获物种资源量组成差异较大.③环境DNA调查次数约占传统渔获物调查次数的几十至几百分之一,采样时间不足努力量最少的渔获物调查的1%,检测到的鱼类种数为传统调查总数的31%~49%.④安庆采样点位于长江中下游长江江豚密度最高的江段,其环境DNA检出率和序列相对丰度在3个采样点中均最高.研究显示:长江水样环境DNA包含水陆复合生态系统的生物多样性信息,利用水样环境DNA宏条形码可检测不同类群的水生和陆生物种;对于鱼类物种检测,环境DNA宏条形码比传统调查方法效率更高,可对传统调查结果进行补充;环境DNA宏条形码生物多样性检测主要受分子标记体系和核酸序列数据库限制,获取全面的物种多样性和资源量信息需要对检测分析方法进行进一步完善.      

联合改良剂对桉树吸收重金属的作用——以矿区土壤中Cd、Zn和Cu为例

通过5种矿区土壤的盆栽试验,研究了联合改良剂对桉树部分生理指标、根中Cd、Zn和Cu的含量、以及土壤中Cd、Zn和Cu的BCR提取形态的影响,探讨了其对矿区土壤还林的作用.结果表明,与对照相比,施用改良剂后桉树的生物量和叶绿素含量的增幅分别为-7.6%~158.8%和-0.7%~23.1%,丙二醛含量的降幅为2.6%~37.9%,其中部分处理的生物量和丙二醛含量的变化达到显著水平（P<0.05）;土壤中有效态Cd、Zn和Cu的含量分别下降12.5%~63.8%、1.6%~54.7%和16.2%~57.6%,桉树根中Cd、Zn和Cu的含量分别下降3.1%~56.6%、-1.5%~21.3%和9.3%~50.1%.土壤对桉树根中Cd、Zn和Cu的含量有显著影响（P<0.05）,改良剂的配比或施用量无显著影响（P>0.05）,但在5种矿区土壤中G3降低桉树根中Cu含量的效果优于G1和G2;在重金属含量最高的2种土壤中,G1降低桉树根中Cd和Zn含量的效果优于G2和G3.研究表明施用联合改良剂能降低桉树对重金属的吸收,有利于矿区土壤的还林.     

铝合金的实验室盐雾试验腐蚀行为图像特征提取

目的提取铝合金材料在盐雾箱试验中可以表征腐蚀程度的表面形貌图像特征量。方法首先,对采集于表面化学清洗过的试样原始图像进行图像增强等预处理,突出腐蚀部位;其次,基于数字图像处理的分形和小波分解方法提取出分形维数和小波能量特征值。结果与以质量损失量为基础的腐蚀深度特征值进行相关性对比,基于图像分析的特征提取法的准确度和精度比较高。结论该方法可以应用于对试样的腐蚀程度进行定性和定量分析,判断并预测试样的腐蚀速度。     

石墨炉原子吸收光谱法测定海河下游水中痕量镉

原子吸收光谱法直接测定高盐水中痕量镉时，有很大背景吸收和误差。本文采用络合—萃取技术使共存元素与待测元素分离，既消除了基体干扰，又达到了富集作用，使测定结果准确可靠。     

超临界二氧化碳萃取－GC／MS测定土壤中的多环芳烃

本文开发了一种采用超临界二氧化碳萃取土壤中多环芳烃、不须经过纯化步骤，直接可用于ＧＣ／ＭＳ分析的简便、高效的方法。本实验中超临界革取的流体是二氧化碳，改善剂是５％的二氯甲烷／甲醇，萃取温度为１２０℃、压力为３４ＭＰａ。ＧＣ／ＭＳ分析时除了采用外标外，还加入了６种同位素ＰＡＨｓ内标以校正各段ＰＡＨｓ的响应因子。采用本方法成功地测定了我国未开垦森林土壤中的ＰＡＨｓ。     

Microbiological characteristics in a zero-valent iron reactive barrier

Zero-valent iron (Fe⁰)-based permeable reactive barriertreatment has been generating great interest for passivegroundwater remediation, yet few studies have paid particularattention to the microbial activity and characteristics withinand in the vicinity of the Fe⁰-barrier matrix. The presentstudy was undertaken to evaluate the microbial population andcommunity composition in the reducing zone of influence byFe⁰ corrosion in the barrier at the Oak Ridge Y-12 Plantsite. Both phospholipid fatty acids and DNA analyses were usedto determine the total microbial population and microbialfunctional groups, including sulfate-reducing bacteria,denitrifying bacteria, and methanogens, in groundwater andsoil/iron core samples. A diverse microbial community wasidentified in the strongly reducing Fe⁰ environment despitea relatively high pH condition within the Fe⁰ barrier (up topH 10). In comparison with those found in the backgroundsoil/groundwater samples, the enhanced microbial populationranged from 1 to 3 orders of magnitude and appeared to increase from upgradient of the barrier to downgradient soil. Inaddition, microbial community composition appeared to change overtime, and the bacterial types of microorganismsincreased consistently as the barrier aged. DNA analysisindicated the presence of sulfate-reducing and denitrifyingbacteria in the barrier and its surrounding soil. However, theactivity of methanogens was found to be relatively low,presumably as a result of the competition by sulfate/metal-reducing bacteria and denitrifying bacteria because of the unlimited availability of sulfate and nitrate in the site groundwater. Results of this study provide evidenceof a diverse microbial population within and in the vicinity ofthe iron barrier, although the important roles of microbial activity, either beneficially or detrimentally, on the longevityand enduring efficiency of the Fe⁰ barriers are yet to be evaluated.