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.     

种植密度和降水对冬小麦田N2O排放的影响

为研究种植密度对农田N2O排放的影响和探讨N₂O排放季节性波动的原因,于1999～2000年小麦生长季在南京市郊江宁县进行了不同播种量(0、90、180和270kg/hm²)的大田试验.观测分析结果表明:在相同的气象条件和田间管理下,播种～越冬阶段的N₂O排放不受播种量的影响,返青～成熟阶段的N₂O排放通量与播种量成正比.裸地条件下的N₂O排放与播量为90kg/hm²下的排放无明显差异.造成该生长季内N₂O排放季节性波动的主要原因是降水,返青～成熟阶段的N₂O排放通量随观测日前6d的加权平均降水量呈指数增加.     

Body condition and immune response in wild zebra finches: effects of capture, confinement and captive-rearing

Behavioural ecologists attempt to predict fitness in birds from estimates of body condition and immune capacity. We investigated how the stresses associated with capture, confinement and captive-rearing of wild zebra finches (Taeniopygia guttata) affected different elements of the immune system and body condition. Wild birds had higher heterophil:lymphocyte ratios and total leucocyte counts than aviary birds, presumably an outcome of mounting specific resistance to pathogens, but this response diminished significantly within 10 days of confinement. Wild birds had lower phytohaemagglutinin-A (PHA) responses than their aviary-bred counterparts possibly because energetic costs limited a general resistance response. Wild birds were heavier and had higher haematocrits than their aviary counterparts, but had less fat, although just 10 days of captivity significantly increased fat levels. Measures of body condition were of limited use for predicting immune responsiveness. We conclude that the different elements of the immune system and body condition respond independently, and often unpredictably, to many ecological and behavioural stressors.     

Survey of polycyclic aromatic hydrocarbons (PAHs) in arterial street air of Hangzhou

ＩｎｔｒｏｄｕｃｔｉｏｎＰｏｌｙｃｙｃｌｉｃａｒｏｍａｔｉｃｈｙｄｒｏｃａｒｂｏｎｓ (ＰＡＨｓ)ａｒｅｗｉｄｅｓｐｒｅａｄｅｎｖｉｒｏｎｍｅｎｔａｌｐｏｌｌｕｔａｎｔｓｉｎｔｈｅａｔｍｏｓｐｈｅｒｅ.Ｔｈｅｙａｒｅｇｅｎｅｒａｔｅｄｉｎｔｈｅｃｏｍｂｕｓｔｉｏｎ .Ｔｈｅｍａｊｏｒｓｏｕｒｃｅｓｉｄｅｎｔｉｆｉｅｄｉｎｃｌｕｄｅｆｏｓｓｉｌｆｕｅｌｓ,ｖｅｈｉｃｕｌａｒｔｒａｆｆｉｃ,ｉｎｄｕｓｔｒｉａｌｐｒｏｃｅｓｓｅｓ,ｓｍｏｋｉｎｇａｎｄｄｏｍｅｓｔｉｃｈｅａｔｉｎｇ (Ｐｅｌｔｏｎｅｎ ,1995) .Ｔ…     

Effect of long-term fertilization on soil nitrate distribution

ＩｎｔｒｏｄｕｃｔｉｏｎＦｅｒｔｉｌｉｚａｔｉｏｎｉｓｅｓｓｅｎｔｉａｌｆｏｒａｇｒｉｃｕｌｔｕｒａｌｐｒｏｄｕｃｔｉｏｎ ,ｂｕｔｕｎｒｅａｓｏｎａｂｌｙａｐｐｌｙｉｎｇｆｅｒｔｉｌｉｚｅｒｓｃｏｕｌｄｒｅｓｕｌｔｉｎｓｅｒｉｅｓｏｆｐｒｏｂｌｅｍｓｓｕｃｈａｓｃｒｏｐｑｕａｌｉｔｙｄｅｃｌｉｎｅ,ｆｅｒｔｉｌｉｚｅｒｕｓｅｅｆｆｉｃｉｅｎｃｙｄｅｃｒｅａｓｅ,ｒｅｓｉｄｕａｌｆｅｒｔｉｌｉｚｅｒｓｐｏｌｌｕｔｉｏｎ ｇｒｏｕｎｄｗａｔｅｒａｎｄｒｉｖｅｒｓａｎｄｎｉｔｒｏｇｅｎｃｏｎｔｅｎｔｅｄ …     

Partition of polycyclic aromatic hydrocarbons on organobentonites from water

ＩｎｔｒｏｄｕｃｔｉｏｎＴｈｅａｌｕｍｉｎｏｓｉｌｉｃａｔｅｓｈｅｅｔｓｏｆｂｅｎｔｏｎｉｔｅｐｏｓｓｅｓｓａｎｅｔｎｅｇａｔｉｖｅｅｌｅｃｔｒｉｃａｌｃｈａｒｇｅｃｏｍｐｅｎｓａｔｅｄｆｏｒｂｙｉｎｏｒｇａｎｉｃｅｘｃｈａｎｇｅａｂｌｅｃａｔｉｏｎｓ(ｅ.ｇ.,Ｎａ+ａｎｄＣａ2+),ｗｈｉｃｈｓｔｒｏｎｇｌｙｈｙｄｒａｔｅｄｉｎｔｈｅｐｒｅｓｅｎｃｅｏｆｗａｔｅｒ.Ｓｕｒｆａｃｅｐｒｏｐｅｒｔｉｅｓｏｆｎａｔｕｒａｌｂｅｎｔｏｎｉｔｅｃａｎｂｅｍｏｄｉｆｉｅｄｂｙｓｉｍｐｌｅｉｏｎｅｘｃｈａｎｇｅｗｉ…     

Prevention of debris flow disasters on Chengdu-Kunming Railway

ＩｎｔｒｏｄｕｃｔｉｏｎＣｈｅｎｇｄｕ ＫｕｎｍｉｎｇＲａｉｌｗａｙＬｉｎｅｉｓｌｏｃａｔｅｄｏｎｔｈｅｍｏｕｎｔａｉｎｒｅｇｉｏｎｏｆｓｏｕｔｈｗｅｓｔｅｒｎＣｈｉｎａ .Ｔｈｅｒｅａｒｅａｌｏｔｏｆｄｅｂｒｉｓｆｌｏｗｇｕｌｌｉｅｓｓｐｒｅａｄｉｎｇｂｅｓｉｄｅｔｈｉｓｒａｉｌｗａｙｌｉｎｅ .Ｏｎｓｕｍｍｅｒ,ｔｈｅｄｅｂｒｉｓｆｌｏｗｏｆｔｅｎｂｕｒｓｔｓａｎｄｔｈｒｅａｔｅｎｓｔｈｅｓａｆｅｔｙｏｆｔｈｅｒａｉｌｗａｙｔｒａｎｓｐｏｒｔａｔｉｏｎ.Ｉｎ 1981,ａｌａｒｇｅ ｓｃａｌｅｄｅｂｒｉｓ…     

Integrated environmental assessment: an urban perspective

１　ＩｎｔｒｏｄｕｃｔｉｏｎＵｒｂａｎａｒｅａｓｔｈｒｏｕｇｈｏｕｔｔｈｅｗｏｒｌｄａｒｅｉｎｃｒｅａｓｉｎｇｌｙａｆｏｃｕｓｏｆｅｎｖｉｒｏｎｍｅｎｔａｌｃｏｎｃｅｒｎ．Ｔｈｅｙａｒｅｐｌａｃｅｓｏｆｉｎｔｅｎｓｅａｎｄｇｒｏｗｉｎｇｅｃｏｎｏｍｉｃａｃｔｉｖｉｔｙ．Ｉｎｒｅｓｐｏｎｓｅ，ｕｒｂａｎｐｏｐｕｌａｔｉｏｎｓａｒｅｇｒｏｗｉｎｇｒａｐｉｄｌｙ．Ｉｔｉｓｅｓｔｉｍａｔｅｄｔｈａｔ，ｂｙ２０００，ｍｏｒｅｔｈａｎｈａｌｆｔｈｅｗｏｒｌｄ○ｓｐｏｐｕｌａｔｉｏｎｗｉｌｌｌｉｖｅｉｎｔｏｗ…     

Particle monitoring techniques for water treatment applications

１　ＩｎｔｒｏｄｕｃｔｉｏｎＰａｒｔｉｃｌｅｓｉｎｎａｔｕｒａｌｗａｔｅｒｓａｒｅｏｆｔｅｎｔｏｏｓｍａｌｌｔｏｂｅｅｆｆｅｃｔｉｖｅｌｙｒｅｍｏｖｅｄｂｙｔｈｅｓｅｓｅｐａｒａｔｉｏｎｔｅｃｈｎｉｑｕｅｓ．Ｔｈｅｐｒｏｃｅｓｓｏｆｃｏａｇｕｌａｔｉｏｎ／ｆｌｏｃｃｕｌａｔｉｏｎｃａｕｓｅｓｆｉｎｅｐａｒｔｉｃｌｅｓｔｏａｇｇｒｅｇａｔｅｉｎｔｏｌａｒｇｅｒｕｎｉｔｓ（ｆｌｏｃｓ）ｗｈｉｃｈｃａｎｂｅｍｏｒｅｅａｓｉｌｙｓｅｐａｒａｔｅｄ．Ｉｎｗａｔｅｒｔｒｅａｔｍｅｎｔｓａｌｔｓｏｆａｌｕｍｉ…     

海水和海洋沉积物中总磷的测定

系统介绍了海水和海洋沉积物中总磷的测定方法，选用Ｋ２Ｓ２Ｏ８为氧化剂将有关形式的Ｐ转化成ＰＯ＾３－４，连同样品中原有的ＰＯ＾３－４－起用以抗坏到为还原剂的磷钼蓝法测定。方法的精密度６％，回收率为９１％－１０７％。