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1.
Ayumi HASEGAWA Keiji MOCHIDA Toshiko TOMISHIMA Kimiko INOUE Atsuo OGURA 《The Journal of reproduction and development》2014,60(3):187-193
Successful in vitro fertilization (IVF) in mice has been achieved using spermatozoa at concentrations
specifically optimized for the experimental conditions, such as species and source of spermatozoa. Although IVF in mice is
mostly performed using about 80–500 µl drops, it is expected that the number of spermatozoa used for insemination can be
reduced by decreasing the size of the IVF drops. The present study was undertaken to examine the extent to which the number
of spermatozoa used for IVF could be reduced by using small droplets (1 µl). We devised the experimental parameters using
frozen–thawed spermatozoa from C57BL/6 mice in anticipation of broader applications to other mouse facilities. We found that
as few as 5 spermatozoa per droplet could fertilize oocytes (1 or 3 oocytes per droplet), although the fertilization rates
were low (13–15%). Practical fertilization rates (> 40%) could be achieved with frozen-thawed C57BL/6J spermatozoa, which
are sensitive to cryopreservation, when 20 sperm per droplet were used to inseminate 3 oocytes. Even with spermatozoa from a
very poor quality suspension (10% motility), about 25% of oocytes were fertilized. Our calculations indicate that the number
of inseminated spermatozoa per oocyte can be reduced to 1/96–1/240 by this method. In two separate embryo transfer
experiments, 60% and 47%, respectively, of embryos developed to term. Our microdroplet IVF method may be particularly
advantageous when only a limited number of motile spermatozoa are available because of inadequate freezing-thawing or genetic
reasons. 相似文献
2.
基于GIS的老挝-中国边境土地适用性评估 总被引:1,自引:0,他引:1
对老挝-中国边境林地使用变更进行了评估,并提出了茶叶的土地适用性。建立一个综合的基于GIS的分析系统(IGAS),该系统支持老挝-中国边境的林地使用和土地适用性的评估研究。采用多标准分析和系统动态技术评估林地使用和土地适用性,同时预测了潜在的茶叶用地。收集到的土地数据和数据分析表明,整体研究区域为10325.07 km2,当前的森林覆被由1992年的6337.33 km2(占61.38%)减少到2002年的5106.28 km2(占49.46%)。当前的森林主要被转为潜在林地和专门的永久农业用地,甚至国家生物多样性保护区域被转为橡胶种植区。林地使用转变的主要原因是贫穷。为了解决这个问题,根据多标准方法,我们对研究区域中适用茶叶种植土地提出了2种分类方案。 相似文献
3.
Satoh R Kaku A Satomura M Kohori M Noura K Furukawa T Kotake M Takano T Hohdatsu T 《Journal of Feline Medicine and Surgery》2011,13(6):427-435
Feline infectious peritonitis virus (FIPV) can cause a lethal disease in cats, feline infectious peritonitis (FIP). The antibody-dependent enhancement (ADE) of FIPV infection has been recognised in experimentally infected cats, and cellular immunity is considered to play an important role in preventing the onset of FIP. To evaluate the importance of cellular immunity for FIPV infection, monoclonal antibodies (MAbs) against feline interferon (fIFN)-γ were first created to establish fIFN-γ detection systems using the MAbs. Six anti-fIFN-γ MAbs were created. Then, the difference in epitope which those MAbs recognise was demonstrated by competitive enzyme-linked immunosorbent assay (ELISA) and IFN-γ neutralisation tests. Detection systems for fIFN-γ (sandwich ELISA, ELISpot assay, and two-colour flow cytometry) were established using anti-fIFN-γ MAbs that recognise different epitopes. In all tests, fIFN-γ production from peripheral blood mononuclear cells (PBMCs) obtained from cats experimentally infected with an FIPV isolate that did not develop the disease was significantly increased by heat-inactivated FIPV stimulation in comparison with medium alone. Especially, CD8(+)fIFN-γ(+) cells, but not CD4(+)fIFN-γ(+) cells, were increased. In contrast, fIFN-γ production from PBMCs isolated from cats that had developed FIP and specific pathogen-free (SPF) cats was not increased by heat-inactivated FIPV stimulation. These results suggest that cellular immunity plays an important role in preventing the development of FIP. Measurement of fIFN-γ production with the anti-fIFN-γ MAbs created in this study appeared to be useful in evaluating cellular immunity in cats. 相似文献
4.
The conservation of rare species on islands is a special challenge, especially when the islands are faced with increasing human impact. Two endangered wild felids in Japan provide a contrast that is useful for examining conservation strategies. The Iriomote cat (Prionailurus bengalensis iriomotensis) is endemic to Iriomotejima Island in Okinawa, and the Tsushima leopard cat (Prionailurus bengalensis euptilurus) occurs in the Tsushima islands in Japan, although a related subspecies found in the Korean Peninsula. Population size was approximately 100 individuals for each subspecies. While the population of Iriomote cat appears relatively stable, the population and abundance of the Tsushima leopard cat has decreased in recent decades. Because of their small population sizes and restricted habitats both are listed as endangered species/subspecies in Japan’s Red List, and the Iriomote cat is listed as an endangered subspecies in the IUCN Red List. Although both are similar-sized felids living on small islands, their ecological characteristics such as food habit, habitat selection, and density differ. These differences seem to be caused by the climate, the biological environment (the species composition of fauna in each island, and the presence of competitors), and the artificial background. The threats facing these two felids are also similar. Habitat destruction, traffic accidents, and negative influences by introduced species are the principal threats to be addressed for their conservation. At the same time, there are differences in the degree of impact by each threat, in each species’ conservation programs in practice, and in the stages of progress and operation of programs. These differences are partly because they have different ecological features and partly because the social background of each island, such as human population, main industry, and historical relationship between human and wildlife, are quite different. We will compare the ecology and state of these two wild felids and discuss the different conservational situations in relation to felid ecology and human society. 相似文献
5.
Takashi Yamasaki Ayumi Enomoto Atsushi Kato Tadashi Ishii Kazumasa Shimizu 《Journal of Wood Science》2011,57(1):76-84
Arabinoglucuronoxylans (AGXs) isolated from the holocellulose of sugi (Cryptomeria japonica) and hinoki (Chamaecyparis obtusa) contained one 4-O-methyl-d-glucopyranosyluronic acid (4-O-Me-d-GlcAp) residue per 6.2 d-xylopyranose (d-Xylp) residues and one 4-O-Me-d-GlcAp residue per 3.8 d-Xylp residues. These AGXs were subjected to partial acid hydrolysis. Analyses by size exclusion chromatography and electrospray-ionization
mass spectroscopy of the neutral sugar fractions in the hydrolysates showed the presence of xylooligosaccharides having a
degree of polymerization of 2-8 in addition to d-Xyl, suggesting that the AGXs from sugi and hinoki contained unsubstituted chains consisting of at least eight d-Xyl residues. The acidic sugars in the hydrolysates were separated into two series of aldouronic acids composed of 4-O-Me-d-GlcAp and d-Xylp by ion-exchange chromatography. The first series included aldouronic acids from aldobiouronic acid (4-O-Me-d-GlcAp-Xyl) to aldopentaouronic acids (4-O-Me-d-GlcAp-Xyl4). The second series were aldouronic acids composed of two 4-O-Me-d-GlcAp residues and 2-4 d-Xyl residues. In these acidic sugars, the uronic acid side chains were located on two contiguous d-Xyl residues. These facts indicated that AGXs from sugi and hinoki had a structural unit containing two 4-O-Me-d-GlcAp residues on two contiguous d-Xyl residues as well as AGXs from spruce and larch. 相似文献
6.
New Forests - The genus Aquilaria (Thymelaeaceae) is widely used for the production of agarwood, and includes several valuable plantation species in Southeast Asia. To understand appropriate... 相似文献
7.
Uematsu Y Yamaguchi T Koike A Yagihara H Hasegawa D Matsuki N Ono K Washizu T Arai T Bonkobara M 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2008,70(8):845-847
A monoclonal antibody, K9BYU, was generated using Escherichia coli recombinant extracellular domain of canine neural-cell adhesion molecule (N-CAM) as an antigen. Immunoreactivity of K9BYU to insect cell recombinant canine N-CAM was demonstrated by Western blotting using Sf9 insect cells transfected with the canine N-CAM gene. In Western blotting against canine brain tissue, K9BYU detected three isoforms of N-CAM that correspond to three major isoforms of human and mouse N-CAM (N-CAM-120, -140, and -180). From these results, K9BYU was considered to be a useful tool for research of canine N-CAM. 相似文献
8.
Journal of General Plant Pathology - Pyricularia oryzae, which causes blast disease in rice, is a serious fungal pathogen of many cereal crops. Mitochondria have been a promising target to develop... 相似文献
9.
Ken‐ichi Kudo Fuminori Kawabata Toumi Nomura Ayumi Aridome Shotaro Nishimura Shoji Tabata 《Animal Science Journal》2014,85(10):904-909
We isolated chicken taste buds and used a real‐time Ca2+ imaging technique to investigate the functions of the taste cells. With RT‐PCR, we found that isolated chicken taste bud‐like cell subsets express chicken gustducin messenger RNA. Immunocytochemical techniques revealed that the cell subsets were also immunopositive for chicken gustducin. These results provided strong evidence that the isolated cell subsets contain chicken taste buds. The isolated cell subsets were spindle‐shaped and approximately 61–75 μm wide and 88–98 μm long, and these characteristics are similar to those of sectional chicken taste buds. Using Ca2+ imaging, we observed the buds' response to 2 mmol/L quinine hydrochloride (a bitter substance) and their response to a mixture of 25 mmol/L L‐glutamic acid monopotassium salt monohydrate and 1 mmol/L inosine 5′‐monophosphate disodium salt, umami substances. The present study is the first morphological demonstration of isolated chicken taste buds, and our results indicate that the isolated taste buds were intact and functional approaches for examining the taste senses of the chicken using Ca2+ imaging can be informative. 相似文献
10.
Botrytis aclada and Botrytis allii, associated with onion gray-mold neck rot and isolated in Hokkaido, were tested for sensitivity to benzimidazole. Of the B. aclada strains, 59% were highly resistant and the remaining 41% were sensitive; all strains of B. allii were sensitive. Resistant strains were widespread in Hokkaido. We analyzed the sequences of the β-tubulin gene of resistant strains and detected the replacement of glutamic acid (GAG) by lysine (AAG) at codon 198. This is the first report of benzimidazole resistance in B. aclada. This study revealed a difference in fungicide sensitivity between the two Botrytis species.
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