Preimplantation genetic diagnostic protocols for α- and β-thalassaemias using multiplex fluorescent PCR

Haemoglobinopathies including α- and β-thalassaemia are the world's most common class of single gene disorder. Prenatal diagnosis (PND) for β-thalassaemia has been proven to be an effective strategy for controlling the incidence of new cases and is widely used in several countries where the disease is common. Successful preimplantation genetic diagnosis (PGD) protocols for β-thalassaemia have been introduced using restriction fragment length polymorphism (RFLP), single-stranded conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE). However, contamination and allele dropout (ADO) remain an important concern for all of these strategies. In the present study two PGD protocols for detecting β-thalassaemia mutations (codon 41-42 and IVSI-110) and one for α-thalassaemia (SEA mutation) have been designed and tested. These methods contain failsafe mechanisms to reduce the risk of misdiagnosis due to ADO or contamination and utilise multiplex fluorescent PCR (F-PCR). Interestingly, amplification efficiency and ADO were significantly affected by the choice of DNA polymerase and the freshness of the single cells used. The close similarity between the DNA sequences of β-globin and δ-globin was also found to be an important issue that necessitated careful design of primers for the β-globin gene. Copyright © 2001 John Wiley & Sons, Ltd.     

微滤过程中腐殖酸与膜表面黏附特性的试验研究

为了进一步研究微滤过程中腐殖酸对微滤膜的污染机制,通过运用原子力显微镜(AFM),结合自制的PVDF胶体探针考察了不同的pH、离子强度以及钙离子浓度等水质条件下膜-腐殖酸与腐殖酸-腐殖酸之间微观黏附力的变化趋势以及与膜的通量衰减幅度之间的相关性.结果发现,微观黏附力是膜面污染产生的根本原因.由于pH的电性中和以及离子强度的电性屏蔽作用,膜-腐殖酸以及腐殖酸-腐殖酸之间黏附力随着pH的减小或离子强度的增大都呈增大趋势;而随着钙离子浓度的增大,由于少量钙离子的"架桥"作用以及过量时的电性中和作用,两种黏附力先增大后减小.各条件下,膜-腐殖酸与腐殖酸-腐殖酸之间黏附力都呈相同的变化趋势,且与对应污染膜的通量衰减幅度具有正相关性,为更加深入了解膜污染的形成机制提供了一定的理论支持.     

铜陵市河流沉积物中硝化和反硝化微生物分布特征

矿区河流不仅受到矿业活动带来污染物的影响,还受到区域内生活污水及农田退水中氮、磷和有机物的污染.本研究以铜陵市河流表层沉积物为研究对象,采用荧光定量PCR技术分析季节和污染类型对沉积物样品中硝化(amo A)和反硝化(nir S和nir K)功能基因丰度的影响.结果表明,沉积物样品中氨氧化古菌(AOA)和细菌(AOB)基因丰度变化范围分别为1.74×105~1.45×108copies·g-1和1.39×105~3.39×107copies·g-1,AOA平均丰度是AOB的4.39倍;反硝化基因(nir S和nir K基因)丰度变化范围分别为1.69×107~8.55×109copies·g-1和4.45×106~1.51×108copies·g-1,nir S基因平均丰度为nir K基因的28.35倍.沉积物AOA丰度呈现春季和秋季较高,夏季和冬季较低的趋势,而AOB在春季和冬季沉积物中的丰度高于夏季和秋季;反硝化基因丰度则表现为春季(nir S)/秋季(nir K)夏季冬季秋季(nir S)/春季(nir K).矿区周围以重金属为主要污染物的河流沉积物中两种amo A和nir S基因的丰度高于毗邻农田区域主要受到氮、磷及有机物污染的河流,后者的nir K基因丰度更高.     

First application of preimplantation genetic diagnosis to neurofibromatosis type 2 (NF2)

Neurofibromatosis type 2 (NF2) is a dominantly inherited cancer predisposition syndrome that is caused bymutations in the NF2 gene. We report here the first clinical preimplantation genetic diagnosis (PGD) forNF2. A protocol was developed to simultaneously amplify the mutation and a single nucleotide polymorphism (SNP) located within the gene. The mutation and polymorphism were analysed by simultaneous fluorescent single-strand conformation polymorphism (SSCP) on an automated DNA sequencer. The mutation, carried by the male partner, was a single base pair substitution affecting a splice site in intron 4 of the gene. The female partner was infertile due to polycystic ovary syndrome and would require IVF to conceive. The couple was found to be informative at a linked intragenic SNP situated in the 5′ untranslated region of the gene. The SNP was included in the assay to reduce the risk of misdiagnosis due to allele dropout (ADO). The couple underwent three cycles of treatment during which a total of 43 blastomeres were biopsied from 31 embryos. Amplification at both loci was obtained in 35 cells (81%). A total of five embryos were transferred, two in the first cycle, two in the second and one in the third. No pregnancy ensued. The results of the diagnoses indicated that, in this couple, the inheritance of the mutation may be non-Mendelian. Out of a total of 32 embryos tested only four were found not to carry the mutation. The reasons for this apparent skew remain unknown. Copyright © 2002 John Wiley & Sons, Ltd.     

Development of a field well head safety system for use with radiation generating devices

This paper discusses the design, testing and application of a new interlock system for field use with neutron generating bore hole probes at the Idaho National Engineering and Environmental Laboratory. Such probes produce 14 MeV neutrons at a yield up to 3×10⁸ neutrons/s, and present an ionizing radiation hazard to personnel particularly when operated in an unshielded configuration. This interlock system prevents personnel from being exposed to the relatively high radiation fields caused by an unsafe condition when an operating neutron generator probe is withdrawn from a well in the ground. An electromechanical interlock prototype system has been designed, built and tested, and has been found to be effective at preventing this event. The design is simple, is effective in mitigating the hazard, and can be installed in minutes. The system could be modified to interlock radiation-generating devices other than neutron generators that are used in similar environments.     

Determining heavy metals in spent compact fluorescent lamps (CFLs) and their waste management challenges: Some strategies for improving current conditions

From environmental viewpoint, the most important advantage of compact fluorescent lamps (CFLs) is reduction of green house gas emissions. But their significant disadvantage is disposal of spent lamps because of containing a few milligrams of toxic metals, especially mercury and lead. For a successful implementation of any waste management plan, availability of sufficient and accurate information on quantities and compositions of the generated waste and current management conditions is a fundamental prerequisite. In this study, CFLs were selected among 20 different brands in Iran. Content of heavy metals including mercury, lead, nickel, arsenic and chromium was determined by inductive coupled plasma (ICP). Two cities, Tehran and Tabriz, were selected for assessing the current waste management condition of CFLs. The study found that waste generation amount of CFLs in the country was about 159.80, 183.82 and 153.75 million per year in 2010, 2011 and 2012, respectively. Waste generation rate of CFLs in Iran was determined to be 2.05 per person in 2012. The average amount of mercury, lead, nickel, arsenic and chromium was 0.417, 2.33, 0.064, 0.056 and 0.012 mg per lamp, respectively. Currently, waste of CFLs is disposed by municipal waste stream in waste landfills. For improving the current conditions, we propose by considering the successful experience of extended producer responsibility (EPR) in other electronic waste management. The EPR program with advanced recycling fee (ARF) is implemented for collecting and then recycling CFLs. For encouraging consumers to take the spent CFLs back at the end of the products’ useful life, a proportion of ARF (for example, 50%) can be refunded. On the other hand, the government and Environmental Protection Agency should support and encourage recycling companies of CFLs both technically and financially in the first place.     

不同氨氮填加量下沉水植物对沉积物DOM荧光特性的影响

采用三维荧光光谱技术,模拟研究了不同氨氮添加量(0,100,200和400 mg/kg)下沉水植物对湖泊沉积物DOM(溶解性有机质)荧光特性的影响.结果表明:不同氨氮添加量下,沉水植物的存在明显增强沉积物DOM荧光强度,其中对类蛋白质和类氨基酸组分荧光强度影响较大(荧光强度分别增大了20%～77%和27%～56%);而对类腐殖酸和类富里酸组分影响相对较小(荧光强度分别增大了5%～25%和21%～29%). 添加氨氮可增强沉水植物对沉积物DOM荧光强度影响的程度,当添加量为100 mg/kg时,沉水植物生长最旺盛,沉积物DOM中类蛋白质和类氨基酸组分荧光强度增幅最大(分别约77%和56%);即外源氨氮通过增加沉积物蛋白质和氨基酸含量增强了沉积物有机氮的生物有效性;而在其他添加量下,DOM各荧光组分强度增幅不大,即适当添加外源氨氮,可通过促进沉水植物生长增加沉积物中蛋白质和氨基酸等含量,从而提高沉积物有机氮的生物有效性.      

钻井液及其组分对中肋骨条藻的毒性效应（Toxic Effects of Water-Based Drilling Fluid and Its Components on Skeletonema costatum）

为探讨钻井液及其组分的毒性效应,采用荧光技术研究了一种水基钻井液WBF及其7种主要组分(HXJ、PAC141、KPAM、PolyAL、XC、PolyA、Mud)对海洋微藻中肋骨条藻(Skeletonema costatum)的毒性作用,以未加入WBF或其组分的中肋骨条藻的叶绿素荧光强度为参照,计算得到钻井液及其组分对中肋骨条藻不同时间的EC50值.结果表明:WBF的7种单组分中,PolyA对中肋骨条藻的96h EC50=2×103mg·L-1,为有毒组分,PolyAL的96h EC50=3×104mg·L-1,为低毒组分,其余5种组分无毒;WBF的有毒和低毒组分复配时其毒性与作用时间有关,PolyA与PolyAL在24h之内表现为协同作用,之后主要表现为拮抗作用;WBF的毒性主要取决于其有毒组分,而低毒和无毒组分对钻井液的毒性有协同增强作用.