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.     

《Ambio》专刊<酸化水体的生物恢复--北方湖泊恢复研究>前言

20世纪70和80年代,"酸雨"成为一个主要公共政策问题.在加拿大和北欧的斯堪的纳维亚半岛的酸化水体中,生物群落及生境遭到频繁破坏,诸如土著鲑鱼和鳟鱼种群受损乃至灭绝的现象屡见不鲜.     

Comparative Analysis of Ecophysiological Characteristics of Stephanodiscus hantzschii Grun. in the Periods of Its Bloom in Recreational Water Bodies

In the periods of summer and autumn bloom of the Stephanodiscus hantzschii Crun. in recreational water bodies, studies on the vertical distribution of chlorophyll a, its contents per unit biomass, efficiency in using photosynthetically active radiation (E_PhAR), and assimilative activity of microalgae were performed. The results confirmed the existence of two ecophysiological forms of St. hantzschii and provided evidence that both forms are typically autotrophic and can efficiently use low-intensity PhAR for photosynthesis.     

Barriers Beyond the Partners: Bureaucratic and Political Constraints to Implementing Joint Forest Management in Tamil Nadu,India

Whereas past research has treated co-management of common pool resources as if villagers and project implementing authorities were the only relevant actors, numerous external factors beyond the control of these two partners create barriers to successful co-management. This paper draws on discussions with Forest Department officials to examine the influence of these forces on the outcomes of Joint Forest Management (JFM) in Tamil Nadu, India. An empirical inquiry into the operational aspects of JFM indicates the important roles of political parties, powerful people, and other state institutions and functionaries as well as the flow of foreign funding. Further, the strong demand by local people for socio-economic development interventions as opposed to improvement of degraded forests belittles the role of the Forest Department relative to other departments. Numerous other conditioning factors and relationships are explored. The authors call for reforms in public governance to allow better participation of all the actors involved for this participatory management approach to succeed and sustain.     

Plant communities in the zone of gas field development and heavy metal accumulation by them

Heavy metal accumulation by plant communities has been studied in different zones of impact of gas field development. The contents of heavy metals in different blocks of plant communities has proved to depend not only on the location of phytocenosis but also on its species composition. Communities dominated by species of the family Poaceae contain smaller amounts of heavy metals than those dominated by species of the family Asteraceae, especially of the genus Artemisia. The order of precedence in the accumulation of heavy metals in different blocks of plant communities has been revealed.     

Prenatal prediction of childhood-onset spinal muscular atrophy (SMA) in Turkish families

Childhood-onset spinal muscular atrophy (SMA) is one of the most common neurodegenerative genetic disorders. SMN1 is the SMA-determining gene deleted or mutated in the majority of SMA cases. There is no effective cure or treatment for this disease yet. Thus, the availability of prenatal testing is important. Here we report prenatal prediction for 68 fetuses in 63 Turkish SMA families using direct deletion analysis of the SMN1 gene by restriction digestion. The genotype of the index case was known in 40 families (Group A) but unknown in the remaining 23 families (Group B). A total of ten fetuses were predicted to be affected. Eight of these fetuses were derived from Group A and two of these fetuses were from Group B families. Two fetuses from the same family in Group A had the SMNhyb1 gene in addition to homozygous deletion of the NAIP gene. One fetus from Group A was homozygously deleted for only exon 8 of the SMN2 gene, and further analysis showed the presence of both the SMN1 and SMNhyb1 genes but not the SMN2 gene. In addition, one carrier with a homozygous deletion of only exon 8 of the SMN1 gene was detected to have a SMNhyb2 gene, which was also found in the fetus. To our knowledge, these are the first prenatal cases with SMNhyb genes. Follow-up studies demonstrated that the prenatal predictions and the phenotype of the fetuses correlated well in 33 type I pregnancies demonstrating that a careful molecular analysis of the SMN genes is very useful in predicting the phenotype of the fetus in families at risk for SMA. Copyright © 2002 John Wiley & Sons, Ltd.