利用聚合酶链式反应技术研究单粒古代稻谷(公元800年)离体DNA片段的扩增

在东南亚地区已发掘出很多古代稻谷,如果能从这些古稻谷中把DNA提出加以分析,可以得到有关栽培稻的系统分化和地理传播方面的直接信息。古代稻种的基因型相当复杂,必须要能从单粒种子开始分析,方能得到有价值的资料。为此,我们进行了从单粒古稻谷中提取DNA的研究工作。本研究采用通常提取植物组织中DNA的方法。从在日本挖掘出的古代稻谷单粒种子中提取出了50—100ng左右的DNA片段。以这些DNA作为模板,用聚合酶链式反应(Polymerase Chain Reaction,PCR)技术将DNA加以扩增。由其中几个DNA片段初步合成了相当于水稻光敏色素基因的DNA序列。     

Radiocesium interception potential (RIP) of smectite and kaolin reference minerals containing illite (micaceous mineral) as impurity

Cesium-137 (¹³⁷Cs) is strongly adsorbed on clay minerals, especially on illite. The adsorption of Cs⁺ on reference clay minerals, however, has not been fully investigated in relation to the presence of illite. The objective of this study was to clarify the effect of impurities (i.e., illite and vermiculite), present in reference smectite group minerals and kaolin minerals, on the retention of Cs⁺. The clay mineralogy of the reference minerals was characterized by X-ray diffraction (XRD). The radiocesium interception potential (RIP) was measured as an index of the Cs⁺ retention ability of clays. The content of illite in clay was represented by the total potassium (K) content given that illite is a major source of K in the clay fraction. The content of vermiculite in clay was represented by the Cs fixation capacity induced by Cs saturation followed by heating of samples at 110°C. Metabentonite and beidellite gave extremely high RIP values compared with other smectite group minerals, although a peak for illite (at 1.0 nm) was not observed in XRD analysis. The reference smectite and kaolin minerals showed a range of RIP values, even though their RIP values are theoretically zero. The RIP values had a significant positive correlation with the total K content of all the reference clay minerals (r_s = 0.621*). This indicated that the retention ability for ¹³⁷Cs depended more on the content of illite, as impurity, rather than the type of bulk mineral. Hence, the contribution of illite to the magnitude of the RIP was elucidated by the combination of measurement of total K content and XRD analysis.     

Timing of Puberty and Its Relationship with Body Growth and Season in Male Raccoons (Procyon lotor) in Hokkaido

The raccoon (Procyon lotor), indigenous to North America, has naturalized in Japan as an invasive alien species, having been introduced into the country in the 1970s. In Hokkaido, the northernmost island of Japan, feral raccoons have been increasing in number and spreading throughout the island. The age at the onset of puberty for raccoons is important for estimating individual lifetime reproductive success and population growth. The present study investigated the timing of and potential factors affecting the onset of puberty in male raccoons in Hokkaido. External characteristics and histology of testes were studied in 151 male feral raccoons and in 1 captive juvenile. For the majority of feral yearling raccoons, prepubertal development began in May, and spermatozoa production began in October prior to their second mating season. However, some larger juveniles attained puberty during the juvenile period. The captive juvenile, which was fed throughout the winter, attained puberty only 11 months after birth. These results suggest that if male raccoons can achieve enough body growth before the first mating season, puberty can be attained early. In both juveniles and yearlings, spermatozoa production was only observed after autumn. This timing coincided with the recrudescence of seasonally active spermatogenesis in adult males. Therefore, attaining puberty in male raccoons appears to require both adequate body nutrient development and several environmental factors that control seasonal testicular changes.     

An efficient molecular technique for sexing tiger pufferfish (fugu) and the occurrence of sex reversal in a hatchery population

The tiger pufferfish (fugu) is one of the most important food fishes in East Asia. Since its testes are regarded as a delicacy, sex determination is economically relevant. Previous studies have identified a missense single-nucleotide polymorphism (SNP) in the Amhr2 (anti-Müllerian hormone receptor type II) gene as a strong candidate for a master sex-determining polymorphism. To distinguish genotypic sex efficiently, we developed a high-resolution melting (HRM) assay for this SNP site. By screening 396 fish from two independent crosses reared under controlled conditions, we observed perfect concordance between the SNP genotype and phenotypic sex. Thus, this method holds great potential for use in high-throughput sexing. When analyzing 293 progeny from a third cross reared under unknown conditions, we unexpectedly found that 25 % of phenotypic males exhibited female genotype. These results suggest that environmental factors such as rearing conditions could influence the sex-determination pathway in pufferfish. Alternatively, genetic modifiers might override the signals from Amhr2. This finding raises a concern regarding enhanced stock management of this species, because sex-reversed fish could compromise the sex ratio in subsequent generations. The HRM assay will also be useful for monitoring the degree of sex reversal before release.     

杜鹃对两种远距离的近缘苇莺的巢寄生:一个平行协同进化的案例?(英文)

大杜鹃(Cuculus canorus)通常寄生小型雀形目鸟类。由于杜鹃雏鸟的排他性拱卵或拱雏,导致宿主繁殖失败,因此宿主进化出如卵识别能力的反寄生策略。反过来,杜鹃也同时进化出如模拟宿主卵的寄生策略。于是两者之间的相互作用可形成类似军备竞赛的协同进化关系。在匈牙利,大苇莺(Acrocephalus arundinaceus)是大杜鹃的主要宿主,寄生率可高达约 40-65%。以往被认为是大苇莺一个亚种的东方大苇莺(A. orientalis)在日本的种群也被大杜鹃寄生(25-40%)。通过采用光纤光谱仪量化大杜鹃及其宿主的卵色特征,我们比较了东、西半球大杜鹃对两种苇莺的寄生模拟程度。鸟类视觉模型显示,对于可见光颜色,杜鹃与宿主之间卵色的模拟程度在匈牙利高于日本,但匈牙利杜鹃和宿主均较大的卵色变异程度可导致卵色模拟困难。杜鹃与苇莺卵色的量度差异在日本较低,这被证明是宿主拒卵行为的一个重要影响因素。苇莺对大杜鹃卵的拒绝率在日本(35%)和匈牙利(37%)相似。与匈牙利不同,在日本,宿主对杜鹃寄生的应对策略如拒卵行为,先于杜鹃的反适应对策。我们认为,两种近缘物种大苇莺对杜鹃寄生的不同反寄生行为,代表了协同进化军备竞赛的不同演化阶段。     

Would behavioral thermoregulation enable pregnant viviparous tropical lizards to cope with a warmer world?

Sceloporus lizards depend on external heat to achieve their preferred temperature (T_sel) for performing physiological processes. Evidence both in the field and laboratory indicates that pregnant females of this Genus select body temperatures (T_b) lower than 34 °C as higher temperatures may be lethal to embryos. Therefore, thermoregulation is crucial for successful embryo development. Given the increase in global air temperature, it is expected that the first compensatory response of species that inhabit tropical climates will be behavioral thermoregulation. We tested whether viviparous Sceloporus formosus group lizards in the wild exhibited differences in thermoregulatory behavior to achieve the known T_sel for developing embryos regardless of local thermal conditions. We quantified field active body temperature, thermoregulatory behavior mechanisms (time of sighting, microhabitat used and basking time) and available microhabitat thermal conditions (i.e. operative temperature) for 10 lizard species during gestation, distributed along an altitudinal gradient. We applied both conventional and phylogenic analyses to explore whether T_b or behavioral thermoregulation could be regulated in response to different thermal conditions. These species showed no significant differences in field T_b during gestation regardless of local thermal conditions. In contrast, they exhibited significant differences in their behavioral thermoregulation associated with local environmental conditions. Based on these observations, the differences in thermoregulatory behavior identified are interpreted as compensatory adjustments to local thermal conditions. We conclude that these species may deal with higher temperatures predicted for the tropics by modulating their thermoregulatory behavior.     

Hypofractionated radiation therapy in the treatment of canine thymoma: Retrospective study of eight cases

Thymomas are one of the most common tumors of the cranial mediastinum in dogs; however there is limited information available on the use of radiation therapy for treating this neoplasm. Objectives of the current retrospective observational study were to describe outcomes and side effects of a hypofractionated radiation therapy protocol in a group of dogs with confirmed thymoma. A total of eight dogs were included. To generate individualized treatment plans, we designed the planning target volume according to the limits on mean lung dose and the percentage of the total lung volume exceeding 20 Gy (V20). The total administered dose was 48–49 Gy, with one fraction per week for a total of six to seven fractions. After therapy, two dogs achieved complete responses, two achieved partial responses, and the disease remained stable in two. Two dogs died during the radiation therapy protocol and were not classified. The median mean lung dose and V20 were 6.0 Gy (range: 3.1–15.0 Gy) and 12.4% (range: 2.3–27.5%), respectively. The overall response rate was 50.0%, and the median time to response following treatment initiation was 22 days (range: 14–115 days). Acute and late side effects were common in the skin and/or lung and were self‐limiting or asymptomatic. The median survival time was not reached (range: 8–1128 days) and the 1 year survival rate was 75.0%. Hypofractionated radiation therapy was well tolerated in this sample of dogs with thymoma and may be considered when owners decline surgical treatment or the tumor is deemed unresectable.     

CONTRAST-ENHANCED ULTRASONOGRAPHY FOR CHARACTERIZATION OF CANINE FOCAL LIVER LESIONS

In six normal beagles and 27 dogs with spontaneous focal or multifocal liver lesions, contrast-enhanced ultrasonography using Sonazoid^® was performed. Sonazoid^® is a newly developed second-generation contrast agent with the ability to be used for real-time contrast imaging along with parenchymal imaging. An appropriate protocol for the evaluation of all three phases (arterial, portal, and parenchymal) was established based on the results for normal beagles. By evaluation of the echogenicity of hepatic nodules during the arterial and parenchymal phases it was possible to differentiate malignant tumors from benign nodules with very high accuracy. In 15 of 16 dogs diagnosed as malignant tumors, nodules were clearly hypoechoic to the surrounding normal liver during the parenchymal phase. Additionally, malignant tumors had different echogenicity compared with the surrounding normal liver during the arterial phase in 14 of 15 dogs. In the portal phase, there were no characteristic findings. Contrast-enhanced ultrasonography with Sonazoid^® appears to improve the characterization of canine focal and multifocal hepatic lesions.