Spindle formation and microtubule organization during first division in reconstructed rat embryos produced by somatic cell nuclear transfer

The present study was conducted to demonstrate the spindle formation and behavior of chromosomes and microtubules during first division in reconstructed rat embryos produced by somatic cell nuclear transfer (SCNT) with cumulus cell nuclei. To demonstrate the effect of oocyte aging after ovulation on the cleavage of SCNT embryos, micromanipulation was carried out 11, 15 and 18 h after injection of hCG. SCNT oocytes were activated by incubation in culture medium supplemented with 5 microM ionomycin for 5 min followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) in mR1ECM for 2-3 h. For immunocytochemical observation, the SCNT embryos were incubated with monoclonal anti-alpha-tubulin antibody and then fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Cleavage rates were significantly higher for oocytes collected after 15 and 18 h rather than for those collected 11 h after injection of hCG (56 and 53%, respectively vs. 28%; P<0.05). Premature chromosome condensation occurred before activation of the SCNT oocytes, but adequate spindle formation was only rarely observed. The distribution of microtubules in SCNT embryos after activation was different from those of fertilized and parthenogenic oocytes, i.e., a dense microtubule organization shaped like a ring was observed. Eighteen to 20 h post-activation, most SCNT embryos were in the 2-cell stage, but no nucleoli were clearly visible, which was quite different from the fertilized oocytes. In addition, first division with and without small cellular bodies containing DNA was observed in the rat SCNT embryos in some cases. The present study suggests that reorganization of transferred nuclei in rat SCNT embryos may be inadequate in terms of formation of the mitotic assembly and nucleolar reorganization.     

All-around 3D plant modeling system using multiple images and its composition

In this study, we developed an all-around 3D plant modeling system that operates using images and is capable of measuring plants non-destructively without any contact. During the fabrication of this device, we selected a method capable of performing 3D model reconstruction from multiple images. We then developed an improved SfM-MVS (Structure from Motion / Multi-View-Stereo) method that enables 3D reconstruction by simply capturing images with a camera. The resulting image-based method offers a high degree of freedom because the hardware and software can comprise commercially available products, and it permits the use of one or more cameras according to the shape and size of the plant. The advantages of the image-based method are that 3D reconstruction can be conducted at any time as long as the images are already taken, and that the desired locations can be observed, measured, and analyzed from 2D images and a 3D point cloud. The device we developed is capable of 3D measurements and modeling of plants from a few millimeters to 2.4 m of height using this method. This article explains this device, the principles of its composition, and the accuracy of the models obtained from it.     

Thermal stability of carp G-actin monitored by loss of polymerization activity using an extrinsic fluorescent probe

ABSTRACT:   The thermal stability of carp G-actin was investigated by monitoring loss of actin polymerization ability. To determine the amount of native actin remaining after heat treatment, actin was labeled with a fluorescence reagent, N-(1-pyrene)iodoacetamide. The loss of polymerization ability of carp actin during heat treatment, at between 45 and 55°C, occurred faster than that of chicken actin. The inactivation rate was influenced by concentrations of ATP and Ca²⁺ in solution. With the increase of Ca²⁺ concentration, the inactivation of carp actin was markedly suppressed. Furthermore, the activation energy of the inactivation of carp actin obtained from an Arrhenius plot was similar to that of chicken actin. These results indicated that the thermal instability of carp G-actin was due to the low affinites of ATP and Ca²⁺ for carp actin described in a previous report.     

Plant species identification using fecal DNAs from red-eared slider and Reeves’ pond turtle in agricultural canals for rural ecosystem conservation

Fecal DNA samples from the red-eared slider and Reeves’ pond turtle, suspected pests of lotus root paddies, were used to identify the plant species eaten by these turtles in order to develop a strategy for rural ecosystem conservation. The fecal samples were obtained from young and adult individuals (mostly female) of both species living in agricultural canals surrounding lotus root paddies in Tokushima Prefecture, Japan. The samples were screened for the presence or absence of DNA from nine plant species using PCR and plant species-specific primers for the rbcL gene of chloroplast DNA. In the red-eared slider, our analysis identified seven plant species in the fecal DNA samples of adults and three plant species in those of young individuals. In Reeves’ pond turtle, our analysis identified two plant species from adult fecal samples and one species from those of young individuals. Thus, adult red-eared sliders consume a greater range of plants than young red-eared sliders or Reeves’ pond turtles. Both turtle species, independently of age, consumed lotus plants and were likely to cause feeding damage to lotus roots. Considering the plant species detected in adult red-eared sliders and these plant habitats, we suggest that this adult turtle is likely to travel between the agricultural canals and the lotus root paddies. These findings will help the development of strategies for preventing damage to lotus roots by these turtles; furthermore, they indicate that fecal DNA analysis will be applicable to investigation of the feeding habits of other animal species.     

Yield estimation of 1-year-old asparagus grown using rootstock-planting forcing culture

Rootstock-planting forcing culture was developed in asparagus to harvest spears even during the seasons when the plants become dormant, but the demand for them high. In this study, cumulative hours during which the air temperature remained lower than 5°C, i.e. chilling hours (CHs), were calculated to determine dormancy breakage for asparagus cultures. We also measured CIELab colour values for cut stems immediately before rootstock digging, and determined whether they could be substituted and/or compensated for CHs while evaluating asparagus plant productivity in different low-temperature backgrounds, and obtained regression equations for yield estimation. Asparagus seedlings were cultivated in seven different regions across Japan and brought to the study site for harvesting. Our regression equation based on CHs and rootstock weight for yield estimation had relatively high fitness (adjusted R² = 0.5795). The colour values of cut stalks at rootstock digging can also be used to evaluate their productivity. These values can be useful in regions where CHs cannot be determined, although their effectiveness was slightly lower than that of CHs of areas adjacent to the study sites.     

Recommended doses of medetomidine-midazolam-butorphanol with atipamezole for preventing hypothermia in mice

A non-narcotic anesthetic combination (Me/Mi/Bu) of medetomidine (Me), midazolam (Mi), and butorphanol (Bu) has been recommended as the injectable anesthesia in mice. An original dose of Me/Mi/Bu (0.3/4.0/5.0 mg/kg) has provided sufficient anesthetic duration of 40–50 min in mice. In addition, atipamezole is available for reversal of Me/Mi/Bu anesthesia. As an adverse effect of Me/Mi/Bu anesthesia, however, severe hypothermia has been also observed in mice. In the present study, we investigated 1) the main agent in Me/Mi/Bu to cause of hypothermia, 2) the effects of the differential doses of atipamezole on hypothermia induced by Me/Mi/Bu anesthesia and on the plasma levels of creatinine phosphokinase and transaminases, and 3) those recommended doses for preventing hypothermia induced by Me/Mi/Bu anesthesia in mice. The results suggested that 1) the α₂-agonist medetomidine is most likely to induce hypothermia in mice under Me/Mi/Bu anesthesia, 2) the antagonism of atipamezole within proper dose range is effective in promoting the recovery from Me/Mi/Bu-induced hypothermia, and 3) Me/Mi/Bu at the recommended dose of 0.2/6.0/10.0 mg/kg enable to provide anesthetic effects for 40 min and is more considerable to prevent the hypothermia than that at the original dose of 0.3/4.0/5.0 mg/kg.     

Intrapancreatic accessory spleen mimicking pancreatic insulinoma with intrapancreatic metastasis in a cat

An 11-year-old neutered male Domestic Shorthair cat presented with a 3-month history of hypoglycemia, two episodes of seizure, and intermittent tick-like signs. Serum biochemistry revealed severe hypoglycemia associated with high insulin concentrations. Dynamic abdominal computed tomography (CT) indicated two pancreatic masses, which were enhanced most during the late arterial phase but had different degrees and variations of attenuation. Partial pancreatectomy was performed. Histopathology and immunohistochemistry confirmed that one mass was an insulinoma and the other was an ectopic splenic tissue, consistent with the differences in imaging findings. When an intrapancreatic lesion with hyper-attenuation on dynamic abdominal CT is detected, not only insulinoma or metastasis of malignancies but also intrapancreatic accessory spleen (IPAS) should be considered as differential diagnoses.     

Analysis of QTL for Seed Dormancy and Their Response to Dry Heat Treatment in Rice(Oryza sativa L.)

Quantitative trait loci(QTL)controlling seed dormancy in rice were identified using recombinant inbred lines(RILs)population derived from the cross between a japonica variety Kinmaze and an indica variety DV85. Seeds of two parental cultivars and each RIL were harvested in 35d after heading.. The germination percentage of these seeds at 30℃for 7 days were measured as the degree of seed dormancy.. QTL analysis was performed with Windows QTL Cartographer 1.13a program by composite interval mapping.. A total of four QTL for seed dormancy were detected on chromosome 2(two regions),5 and 11,respectively.Phenotypic variation explained by each QTL ranged from 8.37 to 17.40%.. Responses of such loci to a dormancy-breaking treatment with dry heat were further detected. The results showed that two alleles of qDOR-2-1 and qDOR-5 from DV85 as well as the allele of qDOR11 from Kinmaze increased the seed dormancy,which seemed to be easily broken by dry heat treatment. Such loci of seed dormancy may be applied to rice genetic improvement.The allele of qDOR-2-2 from DV85 increased the seed dormancy,which could not be broken by dry heat treatment.     

自溶法回收鱼类加工废弃物中蛋白质成分(英文)

鱼头和鱼内脏为鱼类在加工过程中废弃的主要组织,其中含有丰富的蛋白质和脂肪等有益成分,有必要进行回收利用。双鳍舵鲣鱼的头部和内脏约占其鱼体重量的30％,富含蛋白质(14.4％-22.6％)。为了将这些废弃物中的蛋白质进行回收,分别采用双鳍舵鲣鱼内脏本身含有的内源性蛋白酶(自溶)和四种外源性的商业蛋白酶Papain W-40(E1),Pancidase NP-2(E2),Sumichiimu FP(E3)和Protease M(E4)对废弃物中的蛋白质进行水解(50℃,1～4h),考察自溶和不同酶解方法对蛋白质的回收率以及对回收液氨基酸组成的影响。结果显示,自溶法对废弃物中蛋白质的回收效果最好。当水解温度为50℃,水解时间为2h时,自溶法的蛋白质回收率最高,达到76％;其次为E2酶解法,回收率达到70％;E3酶解法效果最差,回收率为63％。进一步研究表明,自溶回收液(AE)的氨基酸总量也最高,达到11756 mg/100g,而采用E1、E2、E3和E4水解方法得到的酶解回收液(EE1、EE2、EE3和EE4)中氨基酸含量分别仅为10294、10642、8750、10121 mg/100g。感观评定结果表明,自溶回...     

Binding profiles and cytokine-inducing effects of fish rhamnose-binding lectins on Burkitt’s lymphoma Raji cells

Rhamnose-binding lectin (RBL) is one of the animal lectin categories which take part in the innate immune responses of fish. Osmerus lanceolatus lectin (OLL) from shishamo smelt eggs is an RBL composed of two tandem-repeated domains, both of which are considered to be a carbohydrate-recognition domain. SAL, catfish (Silurus asotus) egg RBL composed of three domains, binds to Burkitt’s lymphoma Raji cells through globotriaosylceramide (Gb3) carbohydrate chain and to reduce cell size and growth by altering membrane composition without causing cell death. In this experiment, we tried to compare the binding effects of these two RBLs on Raji cells. Flow cytometric and fluorescence microscopic analyses revealed that OLL also directly bound to and shrunk Raji cells with ten times less reactivity than SAL but reduced cell growth with decreasing cell viability. Anti-Gb3 antibody completely blocked the binding of SAL to Raji cells but not that of OLL. In addition, the direct bindings of OLL and SAL to Raji cells were comparably inhibited by melibiose, but lactose was more effective inhibitor for the binding of OLL than that of SAL. These results suggest that OLL has slightly different cell-binding property compared with SAL and binds not only to Gb3 but also to the other carbohydrate receptor-bearing β-galactoside chains. The quantitative RT-PCR analysis revealed that SAL induced the expression of TNF-α but not of IFN-γ, IL-1β, and IL-10. Thus, SAL-induced cytostatic effect on Raji cells might be partially caused by TNF-α-mediated signaling pathway.