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71.
Single-cell pulsed-field gel electrophoresis (SCPFGE) with dual electrode pairs was developed to detect the early stage of DNA fragmentation in human sperm. The motile sperm were purified by the commonly used density-gradient centrifugation technique and subsequent swim-up. The sperm were embedded in a thin film of agarose containing bovine trypsin (20 μg/mL) and were then lysed. Prior to SCPFGE, proteolysis of DNA-binding components, such as protamine and the nuclear matrix was essential to separate the long chain fibers from the fibrous and granular fragments derived from a single nucleus. The overall electrophoretic profiles elucidated the course of DNA fragmentation. A few large fibrous fragments were observed at the beginning of the process, however, as the fragmentation advanced, the long chain fibers decreased and shortened, and, conversely, the granular fragments increased until finally almost all the DNA was shredded. Although the ejaculate contained sperm with heterogeneous stages, the purified motile sperm exhibited several dozens of uniformly elongated fibers arising from the tangled DNA at the origin, whereas a part of these fibers gave rise to fibrous fragments beyond the tip of the elongated fibers, and their numbers and sizes varied among the sperm. Conventional intra-cytoplasmic sperm injection (ICSI) usually depends on intra-operative light microscopic observation to select a sperm for injection. The present results revealed that sperm motility could not give full assurance of DNA integrity. SCPFGE is likely to serve an important role in the preoperative differential diagnosis to determine the competence of the sperm population provided for injection. 相似文献
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Miyazaki H Oyama F Wong HK Kaneko K Sakurai T Tamaoka A Nukina N 《Biochemical and biophysical research communications》2007,361(1):43-48
Processing of APP by BACE1 plays a crucial role in the pathogenesis of Alzheimer disease (AD). Recently, the voltage-gated sodium channel (Nav) β4 subunit (β4), an auxiliary subunit of Nav that is supposed to serve as a cell adhesion molecule, has been identified as a substrate for BACE1. However, the biological consequence of BACE1 processing of β4 remains illusive. Here, we report the biological effects of β4 processing by BACE1. Overexpression of β4 in Neuro2a cells promoted neurite extension and increased the number of F-actin rich filopodia-like protrusions. While coexpression of BACE1 together with β4 further accelerated neurite extension, the number of filopodia-like protrusions was reduced. Overexpression of C-terminal fragment of β4 that was generated by BACE1 (β4-CTF) partially recapitulated the results obtained with BACE1 overexpression. These results suggest that the processing of β4 by BACE1 regulates neurite length and filopodia-like protrusion density in neurons. 相似文献
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Sumiko Hara Chisato Kaneko Hiroatsu Matsumoto Takeshi Nishino Tadao Takeuchi Takeo Mori 《Nucleosides, nucleotides & nucleic acids》2013,32(2-4):571-582
Abstract Novel β-D-ribofuranosides having a 5-substituted imidazo [4,5-d] [1,3]thiazine ring, including the S6-congener 3 of oxanosine 2, were synthesized for screening their anticancer and antiviral activities. 相似文献
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The injurious effect of pisatin on the plasma membrane of pea 总被引:4,自引:0,他引:4
Shiraishi Tomonori; Oku Hachiro; Isono Mitsuko; Ouchi Seiji 《Plant & cell physiology》1975,16(5):939-942
The main cause of wilting which occurs in pea leaves heavilyinfected with powdery mildew was suggested to be due to theinjurious effect of pisatin, a defensive antifungal substanceproduced by the host leaves, which affects the plasma membraneof the same host cells. (Received May 29, 1975; ) 相似文献
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Furihata Ryuji Kizuki Jun Yamano Yuya Mizoguchi Yasuhide Nakajima Suguru Nagai Kou Kaneko Yoshiyuki Yamada Koju Suzuki Masahiro Uchiyama Makoto 《Sleep and biological rhythms》2020,18(2):155-157
Sleep and Biological Rhythms - A 39-year-old married woman was referred to a psychiatry outpatient clinic for treatment of major depressive disorder (MDD). The dose of zolpidem she had been taking... 相似文献
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