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31.
Takafumi SUZUKI Naohito NISHII Satoshi TAKASHIMA Tatsuya MATSUBARA Atsushi IWASAWA Hirofumi TAKEUCHI Kohei TAHARA Tatsuyuki HACHISU Hitoshi KITAGAWA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(11):1379-1383
Polyclonal immunoglobulin (Ig) G autoantibodies against insulin have been
identified in sera of healthy cats. We purified and fractionated insulin-binding IgGs from
cat sera by affinity chromatography and analyzed affinity of insulin-binding IgGs for
insulin and their epitopes. Following the passing of fraction A, which did not bind to
insulin, insulin-binding IgGs were eluted into two fractions, B and C, by affinity
chromatography using a column fixed with bovine insulin. Dissociation constant (KD) values
between insulin-binding IgGs and insulin, determined by surface plasmon resonance analysis
(Biacore™system), were 1.64e−4 M for fraction B (low affinity IgGs) and
2e−5 M for fraction C (high affinity IgGs). Epitope analysis was conducted
using 16 peptide fragments synthesized in concord with the amino acid sequence of feline
insulin by an enzyme-linked immunosorbent assay. Fractions B and C showed higher
absorbance (affinity) of the peptide fragment of 10 amino acid residues at the
carboxyl-terminal of the B chain (peptide No. 19), followed by peptide fragments of 6 to
15 amino acid residues of the B chain (peptide No. 8). Fraction C showed a higher
absorbance to 7 to 16 amino acid residues of the B chain (peptide No. 5) compared with the
absorbance of fraction B. Polyclonal insulin-binding IgGs may form a macromolecule complex
with insulin through the multiple affinity sites of IgG molecules. Feline insulin-binding
IgGs are multifocal and may be composed of multiple IgG components and insulin. 相似文献
32.
Involvement of histone H2B monoubiquitination in the regulation of mouse preimplantation development
Masatoshi OOGA Masataka G. SUZUKI Fugaku AOKI 《The Journal of reproduction and development》2015,61(3):179-184
Histone H2B monoubiquitination (H2Bub1) plays an important role in developmental regulation in various vertebrate species. However, the role of H2Bub1 in mammalian preimplantation development remains unclear. In the present study, we examined the role of H2Bub1 in the regulation of mouse preimplantation development. Based on immunocytochemical analysis using an anti-H2Bub1 antibody, no H2Bub1 signal was detected in the metaphase chromosomes of unfertilized oocytes or the pronuclei of early 1-cell stage embryos, but a weak signal was observed in late 1-cell stage embryos. The signal increased after cleavage into the 2-cell stage, and thereafter a strong signal was observed until the blastocyst stage. To assess the significance of H2Bub1 in the regulation of preimplantation development, RNF20 (an H2B-specific ubiquitin E3 ligase) was knocked down using small interfering RNA (siRNAs). In embryos treated with siRNA, the levels of Rnf20 mRNA and H2Bub1 decreased
at the 4-cell and morula stages. Although these embryos developed normally until the morula stage, only one-third developed into the blastocyst stage. These results suggested that H2Bub1 is involved in the regulation of preimplantation development. 相似文献
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ABSTRACT: Grammistins are hemolytic and ichthyotoxic peptides in the skin secretion of soapfishes and are structurally characterized by their abundance in amphiphilic α-helicity. In the present study, their interaction with lipids and lipid vesicles as well as antibacterial activity were examined using four grammistins (Gs 1 and Gs 2 from Grammistes sexlineatus and Pp 1 and Pp 3 from Pogonoperca punctata ). The hemolytic activity of grammistins was inhibited by phospholipids but not by cholesterol. Moreover, grammistins released carboxyfluorescein entrapped within liposomes made of phosphatidylcholine. In contrast, grammistins were found to have antibacterial activity with a broad spectrum against nine species of bacteria, including both Gram-negative and Gram-positive groups. The potency of their antibacterial activity was not related to that of hemolytic activity, suggesting that grammistins bind to membrane phospholipids but lyse erythrocyte and bacterial membranes via different mechanisms. Conclusively, grammistins are new members of the family of cell non-selective membrane-lytic peptides with amphiphilic α-helices, being similar to pardaxins, which are secreted from the skin of soles, and to melittin, which is derived from bee venom. 相似文献
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Summary A combination of compatible second pollinations and embryo rescue was applied for systematic production of true tetraploid hybrids from crosses between disomic tetraploid Solanum acaule and tetrasomic tetraploid potato, S. tuberosum. Several genotypes of tetraploid potatoes were pollinated with S. acaule, and the compatible second pollinations were made on the following day, with a genotype of S. phureja, IvP 35 to promote fruit development. Embryo rescue was carried out in 21 families, 14 to 27 days after the first pollination. A total of eight plants were obtained from the embryo rescue and their chromosome numbers were counted in the root tips. Three of the eight plants were identified as tetraploid, and five others as diploid. Morphology, isozyme banding patterns, and pollen stainability, as well as potato spindle tuber viroid (PSTVd) resistance, indicated the hybrid nature of the three plants. This is the first report of successful tetraploid hybrid production between disomic tetraploid S. acaule (4x) and tetrasomic tetraploid potatoes. Seed set from the crosses between one of hybrids and diploid potatoes indicated workable levels of both male and female fertility for introgression of valuable genes from S. acaule into the cultivated potato gene pool. The methodology used may be applied to other disomic tetraploid tuber-bearing Solanum species and with some modifications also to distantly related solanaceous species and genera. 相似文献