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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
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Reduction of transport stress of ayu by obligated schooling 总被引:3,自引:0,他引:3
KEI'ICHIRO IGUCHI FUMINARI ITO KOGI OGAWA NAOTO MATSUBARA TAIGA YODO TSUGIKO YAMASAKI 《Fisheries Science》2002,68(4):849-853
We investigated stress responses of ayu Plecoglossus altivelis under the hypothesis that overcrowding during transport can be a stressor and the stress predisposes fish to disease by decline in immunocompetence. The extent of stress response was determined under different fish densities. As inferred from the serum concentration of cortisol 1 day after the treatment, the density at more than 23 kg/m3 , which was within the range of normal transporting procedures, acutely evoked intense stress responses. To examine a possible way to reduce the stress response of fish under crowding conditions, we compared two fish groups established in water tanks with circulating (whirling) and turbulent currents at the same density. The cortisol levels 1 day after the treatment were 2.5-fold higher in turbulent current than in whirling current. Fish in whirling current formed a school, while those in turbulent current aggregated with each other. Schooling of fish induced by water flow may reduce their stress responses during transport. A transport system with a directional water flow should keep fish in good condition without reducing the number of fish contained. 相似文献
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Yuko MATSUBARA Takahiroi TAGAMI Daisuke MATSUNAGA Akiko SANO Kimiaki MARUYAMA 《Animal Science Journal》2011,82(3):428-433
In our continuing effort to generate transgenic chickens, sonoporation was chosen to insert an exogenous gene into the chicken genome. An EGFP expression vector (pCAG‐EGFPac) and microbubbles were injected into the central disc of stage‐X blastoderm or the germinal crescent of stage‐4 embryos, followed by ultrasonic vibration. Nineteen chicks out of 108 treated embryos hatched, six females and six males out of these 19 chicks grew to sexual maturity and two females and three males lived for 3 years. Genomic DNA from 17 out of 35 gonads from embryos and chicks that died before sexual maturity was EGFP‐positive by PCR. No EGFP sequence was detected in the genomic DNA of 322 embryos from six sexually mature females and the semen from four sexually mature males by PCR. When genomic DNA was obtained from various tissues of five 3‐year‐old chickens, the EGFP sequence was amplified from the genomic DNA of the breast muscle of a female (No. 85). The above sequence was subjected to DNA sequencing and verified to be the EGFP sequence. These results showed that sonoporation is an effective tool for the transduction of exogenous genes into chicken embryos for the generation of transgenic chickens. 相似文献
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Hiroshi KAGAMI Jun YASUDA Takahiro TAGAMI Mitsuru NAITO Yuko MATSUBARA Takashi HARUMI Takamasa NOGUCHI Yasuhiro YAMAMOTO Taikai TAKAHASHI Junichi MATSUYAMA Hiroyuki KOMATSU Tamao ONO 《Animal Science Journal》2002,73(6):453-456
Effect of the strain combination of the donor and recipient on production efficiency of W‐bearing sperm in mixed‐sex chimeric testes was analyzed. The combinations of the donors and recipients were White Leghorn (WL) and Rhode Island Red (RIR), and vice versa. Generated mixed‐sex chimeras that had the male phenotype at sexual maturity were classified into four groups: (1) a female WL donor and a male RIR recipient; (2) a male WL donor and a female RIR recipient; (3) a female RIR donor and a male WL recipient; (4) a male RIR donor and a female WL recipient. The mean number of W‐bearing sperm detected by in situ hybridization among 10 000 sperm observed were 147, 165, 30 and 45 in groups 1, 2, 3 and 4, respectively. The numbers in groups 1 and 2 were both significantly higher than those of groups 3 and 4 (P < 0.05). The combination of a WL donor and a RIR recipient produced W‐bearing sperm more efficiently than the reverse combination. 相似文献
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Daichi IJIRI Akito SAEGUSA Tomoko MATSUBARA Yukio KANAI Miho HIRABAYASHI 《Animal Science Journal》2012,83(6):504-509
Chicks (Gallus gallus domesticus) show considerable growth of skeletal muscle during the neonatal period. The in vivo gene transfer method is useful for studying gene function and can be employed to elucidate the molecular mechanisms of skeletal muscle growth in chicks. We evaluated the following conditions for gene transfer to the skeletal muscle of neonatal chicks by electroporation: (i) voltage; (ii) age of the chick; (iii) plasmid DNA injected amount; and (iv) duration of gene expression. The results obtained from this study indicate that the most efficient gene transfer condition was as follows: 75 µg of plasmid DNA encoding β‐galactosidase was injected into the gastrocnemius muscle of chicks at 4 days of age electroporated at 50 V/cm. In addition, peak transferred gene expression was observed from 3 days to 5 days after electroporation. Our results provide optimal electroporation conditions for elucidating the gene function related to skeletal muscle growth and development in neonatal chicks. 相似文献
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Komei HOTTA Takayuki WATANABE Chiho KISHIDA Yukio NAKAMURA Nobuyuki OHKUBO Takahiro MATSUBARA Shinji ADACHI Kohei YAMAUCHI 《Fisheries Science》2003,69(3):555-562
ABSTRACT: Because blood vitellogenin (Vg) has been considered a biomarker for environmental estrogens, the basal levels of Vg and 17β-estradiol (E2 ) were determined in male Japanese whiting reared under natural conditions. Serum levels of Vg and E2 were measured and gonadal development was assessed by gonadosomatic index (GSI) and histological observation in 8–10 male fish at monthly intervals throughout the annual reproductive cycle. Serum E2 was <60 pg/mL throughout the study period. In contrast, serum Vg exhibited seasonal changes: serum levels of Vg gradually increased from April to May (mean 63 ± 13 ng/mL and 124 ± 48 ng/mL in April and May, respectively), and then reached a peak value (mean 352 ± 68 ng/mL) in June. Thereafter, serum Vg gradually decreased, reaching undetectable levels (<50 ng/mL) in October. Serum levels of Vg tended to increase in the male fish in which the GSI was >1%. Histological observation revealed that testes in such male fish were in active spermatogenesis and then all of the testes of male fish in which serum Vg decreased to ND levels were regressed. These results suggest that Vg productive potency (sensitivity to estrogens) may increase in the spermatogenic stage, resulting in production of Vg in response to very low levels of natural or xenobiotic estrogens. 相似文献