Rać Monika Ewa
Z Katedry i Zakladu Biochemii i Chemii Pomorskiej Akademii Medycznej w Szczecinie, al. Powstańców Wielkopolskich 72, 70-111 Szczecin.
Ann Acad Med Stetin. 2003;49:63-77.
The aim of this work was to determine the bioaccumulation of fluoride and some metals (Cu, Zn, Pb) in tissues of snails under strictly controlled conditions expecting with this approach to verify the hypothesis that snails are suitable for the monitoring of environmental hazards. Additionally, the toxicity of fluorides administered orally on the energy balance of the snail's foot was investigated basing on concentrations of nucleosides, nucleotides and their products measured with high-performance liquid chromatography (HPLC). Two parallel snail cultures were started. The effect of dose on tissue levels of fluoride and metals was studied in the first part of the experiment. The second part served to study the effects of fluoride on energy metabolism of foot muscle (Tab. 1). Quantitation of fluoride and metal levels was done in soft tissues (foot, hepatopancreas) and shells of snails. Qualitative and quantitative analysis of purine compounds was performed in slices of foot. Fluoride concentrations in pulverized shells were measured using an ion-selective electrode. Gas chromatography served to determine fluoride concentrations in soft tissues (hepatopancreas and foot). Concentrations of metals were determined spectrophotometrically. Fluoride and metal content was calculated basing on weight of the pulverized sample. Purines were measured in foot muscle slices with high-performance liquid chromatography (HPLC). Concentrations were adjusted for protein content of sample. Concentrations of the following nucleosides, nucleotides and their products were determined: ATP, ADP, AMP, Ado (adenosine), GTP, GDP, GMP, Guo (guanosine), Hyp (hypoxanthine), IMP, Ino (inosine), Xan (xanthine), Urd (uridine), UA (uric acid), NAD+, and NADP (Fig. 1.). Statistical analysis was done with non-parametric test of Kruskal-Wallis, Mann-Whitney U-test and Spearman Rank Correlation Coefficient.
本研究的目的是在严格控制的条件下,测定蜗牛组织中氟化物和一些金属(铜、锌、铅)的生物累积情况,期望通过这种方法验证蜗牛适合用于监测环境危害这一假设。此外,基于高效液相色谱(HPLC)测定的核苷、核苷酸及其产物浓度,研究口服氟化物对蜗牛足部能量平衡的毒性作用。启动了两个平行的蜗牛培养实验。在实验的第一部分研究剂量对氟化物和金属组织水平的影响。第二部分用于研究氟化物对足部肌肉能量代谢的影响(表1)。对蜗牛的软组织(足部、肝胰腺)和贝壳中的氟化物和金属水平进行定量分析。对足部切片进行嘌呤化合物的定性和定量分析。使用离子选择性电极测量粉碎后贝壳中的氟化物浓度。气相色谱法用于测定软组织(肝胰腺和足部)中的氟化物浓度。通过分光光度法测定金属浓度。根据粉碎样品重量计算氟化物和金属含量。用高效液相色谱(HPLC)测量足部肌肉切片中的嘌呤。根据样品蛋白质含量调整浓度。测定以下核苷、核苷酸及其产物的浓度:ATP、ADP、AMP、Ado(腺苷)、GTP、GDP、GMP、Guo(鸟苷)、Hyp(次黄嘌呤)、IMP、Ino(肌苷)、Xan(黄嘌呤)、Urd(尿苷)、UA(尿酸)、NAD + 和NADP(图1)。采用Kruskal - Wallis非参数检验、Mann - Whitney U检验和Spearman秩相关系数进行统计分析。