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Novel application of small domestic herbivora raised by discarded recycling resources

废弃回收资源培育小型家养食草动物的新用途

基本信息

批准号：
07556129
负责人：
MANABE Noboru
金额：
$ 2.82万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

A novel application method of small domestic herbivora raised by discarded recycling resources was developed. Moreover, highly sensitive method to detect the environmental toxicants which induce genetic and teratogenic toxicity to domestic mammals and human was developed. The new assessment method realized non destructive and real time detection. In vivo and in vitro phosphorus-31 and proton nuclear magnetic resonance (NMR) spectra of the intestinal tracts including liver and reproductive organs, ovary and testis, and embryo of experimental and domestic herbivora 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 NMR spectroscopy. Proton NMR experiments were done by a modification of conventional … More 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 25C.In vivo 31P NMR spectra of the steady-state organs within intact animals were obtained using a JNM-alpha-400 FT-NMR spectrometer (JEOL) equipped with 89-mm bore superconducting magnet (9.20 Tesla ; Oxford Instruments). The animals were mounted on a surface coil probe (two-turns, 2.0 cm diameter) which was tuned to 31P at 161.7 MHz and used in conjunction with a ferrite screen composed of strips of computer tape to eliminate 31P NMR signals arising from superficial muscles. 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 3.0 s, 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, and the creatine phosphate peak was used as a positional reference. 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 digestive capacity of small domestic herbivora and for evaluation of reproductive and genetic toxicity in domestic mammals. Less

提出了一种利用废弃再生资源饲养小型草食动物的新方法。此外，还建立了高灵敏度的检测环境毒物对家畜和人类的遗传和致畸毒性的方法。新的评估方法实现了无损、真实的实时检测。在体内和体外的磷-31和质子核磁共振（NMR）光谱的肠道，包括肝脏和生殖器官，卵巢和睾丸，和胚胎的实验和国内草食动物获得使用自制的表面线圈，传统的圆形线圈，和/或成像线圈。在体内观察了代表高能态的β-ATP/无机磷酸盐比值和细胞内pH等。许多环境毒物的鉴定和量化，没有任何提取过程，从这些器官和胚胎使用NMR光谱。质子核磁共振实验是通过修改常规的 ...更多信息技术，并如下进行31 P NMR实验：在NMR检查期间，将样品保持在25 ℃的受控空气温度下的探针室中，使用装备有89-mm孔超导磁体（9.20特斯拉; Oxford Instruments）的JNM-alpha-400 FT-NMR光谱仪（JEOL）获得完整动物体内稳态器官的体内31 P NMR光谱。将动物安装在表面线圈探头（两圈，直径2.0 cm）上，该探头在161.7 MHz下调谐至31 P，并与由计算机磁带条组成的铁氧体屏幕结合使用，以消除由浅表肌肉产生的31 P NMR信号。通过使用源自器官样本的水1H信号调整匀场参数来控制场均匀性。如下设置采集条件以获得完全弛豫信号：脉冲宽度为20微秒，脉冲重复时间为3.0秒，并且累积200次扫描。因此，每次测量花费5-25分钟。在指数乘法之后，对累积的自由感应衰减进行傅里叶变换。通过JEOL软件的计算机积分计算单个光谱峰面积，并将磷酸肌酸峰用作位置参考。通过与球形球中外部参比品（20 mM亚甲基二膦酸盐水溶液）的数据进行比较，将每种化合物的相对量转换为绝对浓度。目前的研究表明，核磁共振光谱是一种新的评估工具，消化能力的小型国内草食动物和评价的生殖和遗传毒性的国内哺乳动物。少