A novel assessment technique of reproductive and genetic toxicity in domestic animals using a nuclear magnetic resonance instrument

用自制表面线圈、常规圆形线圈和/或成像线圈获得了实验动物和农场动物卵巢、肝脏、骨骼肌和胚胎的体内和体外磷-31和质子核磁共振(NMR)谱。体内观察到代表高能状态的β-ATP/无机磷比值和细胞内pH等。使用核磁共振波谱，在没有任何提取过程的情况下，从这些器官和胚胎中识别和量化了许多环境毒物。质子核磁共振实验是通过对常规技术的改进进行的，31P核磁共振实验如下：在核磁共振检查期间，样品保持在探针室中，温度控制在25℃。C.体内完整动物体内稳态器官的31P核磁共振谱使用JNM-Alpha-400FT-NMR谱仪(日本东京JEOL)，配备89 mm口径超导磁体(9.20Tesla；牛津Inst…)更多谣言，马萨诸塞州贝德福德，美国)。动物被安装在一个表面线圈探头(两圈，直径2.0厘米)上，在161.7 MHz下调谐到31P，并与由计算机磁带(Scotch 700；Three M Co.，St.Louis，MO)组成的铁氧体屏幕一起使用，以消除来自浅层肌肉的31P核磁共振信号(7)。通过使用来自器官样本的水1H信号调整垫片参数来控制场均匀度。采集条件设置如下：脉冲宽度为20微秒，脉冲重复时间为3.0次，累计扫描200次。因此，每次测量需要5-25分钟。指数相乘后，对累积的自由感应衰变进行傅里叶变换。用JEOL软件(JEOL)计算各谱峰面积，以磷酸肌酸(CP)峰为位置参照物(0ppm)。通过与球状灯泡中外部参照物(水中20 mM亚甲基二膦酸盐)的数据比较，将每种化合物的相对量转换为绝对浓度。本研究表明，核磁共振波谱是评价实验动物和家养动物生殖毒性和遗传毒性的一种新的评估工具。较少

In vivo and in vitro phosphorus-31 and proton nuclear magnetic resonance (NMR) spectra of the ovary, liver, skeletal muscle and embryo of experimental and farm animals were acquired using a house-made surface coil, conventional round coil, and/or imaging coils. Beta-ATP/inorganic phosphate ratio which represents the high energy state and intracellular pH etc.were observed in vivo. Many environmental toxicants were identified and quantified without any extraction processes from these organs and embryos using a NMR spectroscopy. Proton NMR experiments were done by a modification of conventional techniques, and 31P NMR experiments were performed as follows : During the NMR examination, the sample was maintained in a probe chamber at controlled air temperature of 25. C.In.vivo 31P NMR spectra of the steady-state organs within intact animals were obtained using a JNM-alpha-400 FT-NMR spectromenter (JEOL,Tokyo, Japan) equipped with 89-mm bore super-conducting magnet (9.20 Tesla ; Oxford Inst … More ruments, Bedford, MA,USA). The animal was mounted on a surface coil probe (two-turns, 2.0cm diameter) with was tuned to 31P at 161.7MHz and used in conjunction with a ferrite screen composed of strips of computer tape (Scotch 700 ; Three M Co., St.Louis, MO) to eliminate 31P NMR signals arising from superficial muscles (7). Field homogeneity was controlled by adjusting shim parameters using a water 1H signal derived from the organ sample. Acquisition conditions were set as follows to obtain fully relaxd signals : the pulse width was 20 micro-seconds, the pulse repetition time was 3.0s, and 200 scans were accumulated. Thus, it took 5-25 min for each measurement. After exponential multiplication, the accumulated free induction decays were subjected to Fourier transformation. Individual spectral peak areas were calculated by computer integration with JEOL software (JEOL), and the creatine phosphate (CP) peak was used as a positional reference (0 ppm). The relative amount of each compound was converted to absolute concentration by comparison with the data of an external reference (20mM methylene diphosphonate in water) in a spherical bulb. The present study showed that NMR spectroscopy is a novel assessment tools for assessment of reproductive and genetic toxicity in experimental and domestic animals. Less