The oxidation of octanoic acid by cultured amniotic fluid cells: The effect of cell type and passage number

We have measured the rate of oxidation of [1⁻¹⁴C]octanoate in cultured amniotic fluid (AF) cells at various passages and in AF cell lines with different clonal morphology. It is possible that both the passage number and the cell type may influence the outcome of prenatal diagnosis of fatty acid oxidation defects using this technique. We found that there was no significant difference between the three major AF cell types (epithelial, large epithelial, and fibroblast) when analysed at identical passage number but there was a significant reduction in octanoate oxidation in all cell types with increasing passage. For reliable prenatal diagnosis, cell lines of similar low passage number should be used.     

Arsenate and phosphate interaction in Saccharomyces cerevisiae

In the present study, arsenate（As（V）） and phosphate（P（V）） interactions were investigated in growth, uptake and RNA content in yeast（Saccharomyces cerevisiae）. Yeast grew slowly with As（V） concentrations increasing in the medium. However, the maximal population density was almost the same among different As（V） treatments. It was in the late log phase that yeast growth was aug- mented by low As（V）, which was maybe due to the fact that methionine metabolism was stressed by vitamin B6 deprivation, so As（V） treatments did not affect maximal population density. However, with P （V） concentrations increasing, the maximal population density increased. Therefore, the maximal population density was determined by P （V） concentrations in the medium but not by As （V） concentrations in the medium. Ycflp（a tonoplast transpor） transports As（GS）3 into the vacuole, but arsenic（As） remaining in the thalli was 1.27% with As（V） exposure for 60 h, from which it can be speculated that the percentage of As transported into vacuole should be lower than 1.27%. However, the percentage of As pumped out of cell was 71.49% with As （V） exposure for 68 h. Although two pathways （extrusion and sequestration） were involved in As detoxification in yeast, the extrusion pathway played a major role in As detoxification. RNA content was the highest in the early-log phase and was reduced by As（V）.     

Mosaic trisomy 17 in amniotic fluid cells not confirmed in the newborn

A case of true fetal mosaicism 46,XY/47,XY, + 17 was diagnosed in amniotic fluid cells. After genetic counselling and unsuccessful periumbilical blood sampling the pregnancy continued to term, and a healthy male infant was born. Lymphocytes of the newborn had a normal karyotype. Follow-up of the child at age 18 months showed normal physical and mental development indicating that the trisomic cell line was restricted most probably to the extra fetal tissue.     

Cross-hybridization of the chromosome 13/21 alpha satellite DNA probe to chromosome 22 in the prenatal screening of common chromosomal aneuploidies by fish

In a routine application of commercially available centromeric DNA probes for the prenatal screening of common trisomies involving the autosomes 13, 18, and 21, and sex chromosomes, four cases of discrepancy between fluorescence in situ hybridization (FISH) results and follow-up cytogenetic analysis were observed from a total of 516 cases of amniocentesis. In three of these cases, the results were false negative, and in one false positive. In this case, amniocentesis was performed because of a positive triple test in a 34-year-old woman with previous infertility treatment. The alpha satellite DNA probe for chromosomes 13/21 revealed five signals in 50 per cent of uncultured amniocytes, while standard cytogenetic analysis showed a normal karyotype. FISH analysis on metaphase chromosomes demonstrated the location of the additional signal in the centromeric region of chromosome 22. This additional signal was also present in the centromeric region of chromosome 22 of the mother, providing evidence for a possible inherited polymorphism in chromosome 22 responsible for unspecific hybridization with the alpha satellite probe for chromosomes 13/21 in this case. The observed polymorphism in centromeric regions may contribute to unreliability of the use of the 13/21 alpha satellite probe for prenatal screening by FISH.     

低硒环境中人群对不同形态硒的保留量及其效应的研究

研究了低硒生态环境中有明显早期大骨节病病变的人群服用富硒酵母,亚硒酸钠和肌肉注射硒代二乙酸后体内硒保留量的动态变化。分析了治疗效应与摄入三种形态的硒后硒在体内保留量的关系,并与对照组作了比较。讨论了不同形态硒的生物活性。研究结果是三种形态的硒对早期大骨节病病变均有非常显著的疗效,再一次证明低硒生态环境是大骨节病流行的基础。     

黄孢原毛平革菌酵母双杂交cDNA文库的构建及应用

作为研究木质素降解系统的模式微生物黄孢原毛平革菌,迄今还没有蛋白-蛋白相互作用方面的报道.本研究利用酵母双杂交技术构建了低氮培养基中培养2d和3d的酵母双杂交cDNA文库,分别得到2.5×105和3.0×105个重组子.以14-3-3蛋白为钓饵筛选3d cDNA文库,在SD/-Ade/-His/-Leu/-Trp缺陷培养基平板上共获得了约600个单克隆,进一步分析发现,有30个克隆可激活3个报告基因.分离自这些克隆中的质粒DNA能转化大肠杆菌.利用HaeⅢ和Sau3AⅠ限制酶切分析可将它们分为4类.序列测定和同源分析结果表明,其中一类为14-3-3蛋白基因,提示14-3-3蛋白可形成二聚体,另外3类在GenBank没有明显同源的基因.通过SBASE网站预测,编号为Y2H333的克隆含有一个OB-fold核酸结合结构域(OB-fold nucleic acid binding domain).这些研究结果表明,所构建的黄孢原毛平革菌cDNA酵母杂交文库是成功的,14-3-3蛋白在黄孢原毛平革菌中能以蛋白-蛋白相互作用的形式发挥功能.图4表1参28     

包埋固定化微生物法处理含油废水研究

本研究通过包埋固定化微生物法固定除油菌（Y1’菌），用于处理含油废水，并以水体中乳化油去除率为指标考察了影响乳化油降解的各种因素。选用聚乙烯醇（PVA）-海藻酸钠（SA）复配作为包埋固定化载体材料，制备成固定化微生物小球（IMB），通过实验优化了IMB制备的工艺条件。连续批次除油实验结果表明，在25-40℃，固液比1：10，HRT为6h的条件下，进水油含量在20—50mg／L，乳化油去除率可达85％-90％，出水油含量低于5mg／L。     

不同pH条件下酵母菌处理高含油废水的研究

在使用混合酵母菌菌株处理高浓度含油废水中,研究了序批式反应器(SBR)内酵母菌在不同起始pH条件下对废水的处理效果.通过与pH中性或碱性条件比较发现,pH 4～5的条件在酵母菌的发泡抑制、菌体生长以及COD/油去除等方面均显示出更好的效果,pH 5是系统运行的最佳条件.长达100d的SBR连续运行试验表明,系统可以在pH=5条件下长期、稳定运行.     

酵母基因报道系统检测环境二恶英类似物的污染

利用一套酵母基因报道检测系统，建立和标化了对二恶英类标准样品测定的方法，证明此系统可检测ng级的二恶英及其类似物，具有简单、快速的的特点．对广州市区工业和生活污水排放较为集中的水体进行的测定显示，二恶英及其类似物在部分地区超标，而各区经处理的自来水均无超标污染．对小鼠急性毒性实验的血清样品的测定同样证明系统能够测定未浓缩处理的复杂样本，具有一定的应用价值．图4参10     

Demonstration of spontaneously dividing male fetal cells in maternal blood by negative magnetic cell sorting and fish

We investigated a case of massive feto-maternal bleeding by using negative magnetic cell sorting (MACS) and fluorescent in situ hybridization (FISH). A 37-year-old pregnant woman had an uncomplicated amniocentesis for advanced maternal age at 16 weeks' gestation. The fetal karyotype was 46, XY. At 19 weeks' gestation, she had a minor car accident and slight vaginal bleeding. A subsequent Kleihauer-Betke test showed a 140 ml feto-maternal haemorrhage. Serial sonographic examinations indicated a normal fetus and placenta. We performed FISH analysis on maternal peripheral blood at 25 weeks. Anti-CD45 and MACS were used to deplete maternal leucocytes, enriching the proportion of fetal nucleated erythrocytes present. The isolated cells were analysed by using dual-colour FISH with X and Y specific probes. Approximately 65 800 nucleated cells were obtained after MACS depletion. A total of 234 cells were analysed by FISH. The results revealed that 70 of the nucleated cells (30 per cent) were male with one X and one Y signal. Among these cells, six male metaphases were observed in spontaneously dividing cells.