蚕豆根细胞慧星试验对西河水诱变性检测

本文采用新建立的蚕豆根细胞慧星试验对南充市西河三个污染断面河水污染物的DNA损伤作用进行了监测。结果表明,所监测的三个点均存在较明显的诱变剂／致癌剂污染;新建立的蚕豆根细胞慧星试验的灵敏度足以直接检测水体环境致突变／致癌物的污染。     

Lipophilic organic pollutants induce changes in phospholipid and membrane protein composition leading to Vero cell morphological change

Membrane damage related to morphological change in Vero cells is a sensitive index of the composite biotoxicity of trace lipophilic chemicals. However, judging whether the morphological change in Vero cells happens and its ratio are difficult because it is not a quantitative characteristic. To find biomarkers of cell morphological change for quantitatively representing the ratio of morphological changed cell, the mechanism of cell membrane damage driven by typical lipophilic chemicals, such as trichlorophenol (TCP) and perfluorooctanesulphonate (PFOS), was explored. The ratio of morphologically changed cells generally increased with increased TCP or PFOS concentrations, and the level of four major components of phospholipids varied with concentrations of TCP or PFOS, but only the ratio of phosphatidylcholine (PC)/phosphatidylethanolamine (PE) decreased regularly as TCP or PFOS concentrations increased. Analysis of membrane proteins showed that the level of vimentin in normal cell membranes is high, while it decreases or vanishes after TCP exposure. These variations in phospholipid and membrane protein components may result in membrane leakage and variation in rigid structure, which leads to changes in cell morphology. Therefore, the ratio of PC/PE and amount of vimentin may be potential biomarkers for representing the ratio of morphological changed Vero cell introduced by trace lipophilic compounds, thus their composite bio-toxicity.     

Effect of wetland plant fermentation broth on nitrogen removal and bioenergy generation in constructed wetland-microbial fuel cells

● Fermentation broth facilitates N removal and energy yields in tertiary CW-MFC. ● Carbon sources are preferred for nitrogen removal over electricity generation. ● A mutual promotion relationship exists between acetic and humic acid in N removal. ● Humic acid boosts the abundances of functional genes relate to nitrogen metabolism. Constructed wetlands (CWs) are widely used as a tertiary treatment technology, and the addition of carbon sources can significantly improve advanced nitrogen removal. However, excessive carbon sources would lead to an increase in the effluent chemical oxygen demand in CWs, and microbial fuel cells (MFCs) can convert these into electricity. In this study, constructed wetland-microbial fuel cells (CW-MFCs) were built to achieve simultaneous nitrogen removal and electricity generation, using wetland plant litter fermentation broths as carbon sources. The total nitrogen removal in the groups with fermentation broth addition (FGs) reached 83.33%, which was 19.64% higher than that in the CG (group without fermentation broth), and the mean voltages in the FGs were at least 2.6 times higher than that of the CG. Furthermore, two main components of the fermentation broths, acetic acid (Ac) and humic acid (HA), were identified using a three-dimensional excitation emission matrix and gas chromatograph and added to CW-MFCs to explore the influence mechanism on the treatment performance. Denitrification and electrogenesis presented the same tendency: Ac&HA > Ac > CG’ (groups without Ac and HA). These results indicate that Ac and HA increased the abundance of functional genes associated with nitrogen metabolism and electron transfer. This study demonstrated that CW-MFC fermentation broth addition can be a potential strategy for the disposal of secondary effluent and bioelectricity generation.     

藻密度手工监测实验室间比对研究

为研究不同实验室藻密度监测数据的可靠性,现场采集了藻密度样品,处理为现场平行样和实验室前处理后平行样2种,分别由5家实验室的6名技术人员开展检测分析。参考相关技术规范要求,结合其他领域相似工作的成功应用案例,采用Z比分数法对分析结果开展藻密度实验室间比对。结果表明,数据的相对偏差均符合技术规范要求;对藻密度进行数据直接比对和对数转换后比对,各实验室对现场平行样和实验室前处理后平行样的检测结果均为合格;分析方法原理导致藻密度手工监测的绝对数值偏差较大,对当前的太湖水华预警工作适用性不高。     

水华蓝藻细胞提取物对实验鼠毒理效应研究进展

蓝藻暴发引发的水华造成了严重的生态灾难和健康危害,蓝藻水华污染因其已成为全球性的环境问题而受到关注。蓝藻产生的藻毒素具有潜在致癌作用,与多种消化道肿瘤的发病率升高相关,其毒理学效应及其健康危害性更是学术界研究和关注的热点。水华蓝藻细胞提取物较之纯藻毒素的毒理学效应有很大不同,研究水华蓝藻细胞提取物对哺乳动物的毒理学效应能为蓝藻水华的健康风险评估提供一定的理论依据。水华蓝藻细胞提取物对实验鼠产生的毒理学效应,如脏器细胞凋亡和对各脏器抗氧化酶防御系统的损害;水华蓝藻细胞提取物对实验鼠遗传毒性和促癌或致癌作用研究是目前研究的热点和难点问题。     

Transgenic Spartina alterniflora for phytoremediation

Perennial monoculture forming grasses are very important natural remediators of pollutants. Their genetic improvement is an important task because introduction of key transgenes can dramatically improve their remediation potential. Transfer of key genes for mercury phytoremediation into the salt marsh cordgrass (Spartina alterniflora) is reported here. S. alterniflora plays an important role in the salt marsh by cycling of␣elements, both nutrients and pollutants, protects the coastline from erosion, is a keystone species in the␣salt marsh supporting a large food web, which in turn supports a significant segment of economy, including tourism, has an impact on cloud formation and consequently on global weather, and is thus an ecologically important species relevant for our life-support systems. Embryogenic callus of S. alterniflora was co-inoculated with a pair of Agrobacterium strains LBA4404 carrying the organomercurial lyase (merB) and mercuric reductase (merA) genes, respectively, in order to co-introduce both the merA and the merB genes. Seven stable geneticin resistant lines were recovered. The presence of merA and merB genes was verified by PCR and Southern blotting. All but one transgenic lines contained both the merA and the merB sequences proving that co-introduction into Spartina of two genes from separate Agrobacterium strains is feasible and frequent, although the overall frequency of transformation is low. Northern blotting showed differences in relative expression of the two transgenes among individual transformants. The steady-state RNA levels appeared to correlate with the phenotype. Line #7 showed the highest resistance to HgCl₂ (up to 500 μM), whereas line #3 was the most resistant to phenylmercuric acetate (PMA). Wild-type (WT) callus is sensitive to PMA at 50 μM and to HgCl₂ at 225 μM.     

根瘤菌全细胞组分的红外光谱特征

利用红外光谱仪记录并分析了36株根瘤菌全细胞组分的红外光谱图，所得图谱分辨率高、重现性好，菌株之间的光谱特征和吸收带的位置、强度各不相同。尤其在指纹区（1700-650cm^-1），不同菌株的图谱有较为明显的差别，利用红外光谱技术可以进行根瘤菌的鉴别，菌株水平的差异能充分表现出来，红外光谱分析在一定程度上可反映根瘤菌的分类地位，但实际应用起来，仍有一定的困难。     

Case report: birth after preimplantation genetic diagnosis of a subtle mutation in SMN1 gene

Spinal muscular atrophy (SMA) preimplantation genetic diagnosis (PGD) has been available since 1998. Protocols are based on the detection of the homozygous deletion of exon 7, which are present in 90–98% of SMA patients. A couple where the woman was a heterozygous carrier of the usual SMN1 Del7 mutation and the man was a heterozygous carrier of pMet263Arg substitution in exon 6 of SMN1 gene was referred for PGD. The usual PGD test being unsuitable for this couple, we developed a novel duplex polymerase chain reaction (PCR)-based PGD test for the detection of the mutation pMet263Arg by allele specific amplification, combined with the amplification of D5S641 extragenic polymorphic marker. PCR conditions were established using single control lymphoblasts and lymphocytes from the pMet263Arg substitution carrier. Amplification was obtained in 100% of the 86 single cells tested, amplification refractory mutation system (ARMS) PCR was specific in 100% of single cells tested and a complete genotype (mutation plus D5S641) was achieved in 88% of them. A PGD cycle was performed successfully and a pregnancy was obtained. An unaffected girl was born and postnatal diagnosis confirmed PGD results. This is the first PGD described for SMA because of another mutation than the major homozygous exon 7 deletion of SMN1. In the future, a similar strategy could be adopted for other subtle mutations of this gene. Copyright © 2006 John Wiley & Sons, Ltd.     

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.