Development of genotyping method for functionally relevant variants of cytochromes P450 in cynomolgus macaques          下载免费PDF全文

Y. Uno  N. Osada  S. Sakurai  N. Shimozawa  T. Iwata  K. Ikeo  H. Yamazaki 《Journal of veterinary pharmacology and therapeutics》2018,41(1):e30-e34

In cynomolgus macaques (Macaca fascicularis), widely used in drug metabolism studies, CYP2C9, CYP2C76, CYP2D6, CYP3A4, and CYP3A5, important drug‐metabolizing enzymes, are abundantly expressed in liver and metabolize cytochrome P450 substrates. CYP2C9 (c.334A>C), CYP2C76 (c.449TG>A), CYP2D6 (c.891A>G), CYP3A4 (IVS3 + 1G>del), and CYP3A5 (c.625A>T) substantially influence metabolic activity of enzymes, and thus are important variants in drug metabolism studies. In this study, a real‐time PCR method was developed for genotyping these variants. The validity of the methods was verified by genotyping two wild type, two heterozygous, and two homozygous DNAs and was used to genotype 41 cynomolgus macaques (from Cambodia, Indonesia, the Philippines, or Vietnam) for the five variants, along with another important variant CYP2C19 (c.308C>T). The CYP2C9 and CYP2C19 variants were found only in Cambodian and Vietnamese animals, while the CYP2C76 and CYP2D6 variants were found only in Indonesian and Philippine animals. The CYP3A4 and CYP3A5 variants were not found in any of the animals analyzed. Mauritian animals, genotyped using next‐generation sequencing data for comparison, possessed the CYP2C19 and CYP2D6 variants, but not the other variants. These results indicated differences in prevalence of these important variants among animal groups. Therefore, the genotyping tool developed is useful for drug metabolism studies using cynomolgus macaques.  相似文献   

Vitrification for bovine embryos with low‐quality grade          下载免费PDF全文

Hiroshi Tajimi  Tsugumi Yamazaki  Syoko Oike  Tatsuyuki Yoshida  Konosuke Okada  Masashige Kuwayama  Hitoshi Ushijima 《Animal Science Journal》2018,89(8):1194-1200

This study was conducted to examine the utility of vitrification for bovine embryos with low‐quality grade, and simple cryoprotectants dilution method for practitioners. In Experiment 1, survival of frozen embryos was compared with that of vitrified embryos using minimum volume cooling (MVC). Then, vitrified embryos were used to confirm the optimum sucrose concentration in Experiment 2. The survival rates of embryos that had been vitrified following diluted cryoprotectants with the one‐step in‐straw method were compared with those of fresh control embryos in Experiment 3. Frozen‐thawed or vitrified‐warmed blastocysts were cultured with TCM‐199 supplemented with 100 μmol/L beta‐mercaptoethanol +5% fetal bovine serum at 38.5°C in an atmosphere of 5% CO₂ in air, their survival after 24 hr were compared. The development to term of fair quality in vivo embryos after vitrification was examined in Experiment 4. Results show that survival rates of frozen‐thawed embryos were lower (p < .05) than that of vitrified‐warmed ones. When vitrified embryos were warmed in 0.3 mol/L sucrose in straws, their survival rate was 100%. The total cell numbers of vitrified‐warmed embryos were comparable to those of fresh control embryos. The six calves from 13 vitrified embryos were delivered in Experiment 4. These results indicate that MVC vitrification following one‐step cryoprotectants dilution is utilized to preserve low‐quality bovine embryos.  相似文献   

Morphological changes and habitat shifts with growth of endangered floodplain fish: Possible adaptations to fluctuating environments     

Masaki Nishio  Kaneaki Edo  Ryosuke Kawakami  Tomonori Kawamoto  Yuji Yamazaki 《Ecology of Freshwater Fish》2020,29(2):197-209

Adaptations that maximise food intake are dependent on food and/or habitat types. However, there have been few studies on the ability of floodplain primary freshwater fish to rapidly change their morphology, especially the internal one, to maximise utilisation efficiency of food resources in response to environmental fluctuations during their life history. We investigated morphological changes of an Acheilognathinae species (Acheilognathus longipinnis) that inhabits creeks in Japan to determine whether body shape variation correlated with environmental characteristics, including water depth, current velocity, vegetation cover and food availability. When shifting from floodwater-level to low-water-level season, the current velocity of the study area increased as the water level decreased, causing a significant increase in periphyton and a significant decline in zooplankton and phytoplankton. Concurrently, during the short period when A. longipinnis grow from the juvenile to the premature stage, changes in internal morphology, that is intestinal elongation, and changes in external morphology such as an increase of body depth were observed. In contrast, intestinal length significantly decreased in the adult stage. Our findings suggest that morphological changes between floodwater-level and low-water-level seasons associated with juvenile to premature development are an adaptation to changes in food availability due to environmental fluctuations. Additionally, the shortening of the intestine from the premature to the adult stage may be an adaptive strategy for reproduction.  相似文献   

Intravaginal administration of estradiol benzoate capsule for estrus synchronization in goats     

Sayaka MATSUMOTO  Tomomi TANAKA  Natsumi ENDO 《The Journal of reproduction and development》2021,67(2):83

Estrus synchronization requires multiple treatments of hormonal drugs, requiring considerable time and cost. The aim of the present study was to develop an estrus synchronization protocol using intravaginal administration of estradiol benzoate (EB) capsules in goats. Two types of capsules were prepared: an EB capsule that melted immediately after administration and a sustained-release (SR) EB capsule that dissolved slowly and reached a peak after 24 h. Goats with functional corpus lutea were intramuscularly treated with prostaglandin F_2α (PG). At 24 h after PG administration, goats were administered 1 mg of EB solution intramuscularly (PG + 24IM; n = 6) or 1 mg of EB capsule intravaginally (PG + 24EB; n = 6). The SR EB capsule was administered intravaginally at the time of PG administration (PG + SR; n = 6). The control group (n = 6) received only PG. All groups showed estrus within 72 h after PG administration. The onset of estrus did not differ significantly between the PG + 24IM and PG + SR groups but was earlier than in the control group. Estradiol concentration in the PG + SR group peaked at 11.5 ± 6.1 h after EB and PG administration. Peak estradiol concentrations were not significantly different between the PG + 24IM and PG + SR groups (78.0 ± 25.8 and 64.0 ± 38.1 pg/ml, respectively), and were higher than the PG + 24EB and control groups (27.3 ± 8.8 and 14.6 ± 6.1 pg/ml, respectively). These results suggest that intravaginal administration of an EB capsule with a sustained-drug release base is applicable for estrus synchronization, as an alternative to intramuscular administration.  相似文献   

Facile synthesis of rubiadin by microwave heating     

Toshiyuki Takano  Tomomi Kondo  Fumiaki Nakatsubo 《Journal of Wood Science》2006,52(1):90-92

Rubiadin (4) (1,3-dihydroxy-2-methylanthraquinone), which is one of the natural dyes from the roots of Rubiaceae, was synthesized from phthalic anhydride (1) and 2,6-dihydroxytoluene (3) by two reaction steps, that is, a Friedel-Crafts reaction with AlCl₃ and a cyclization with H₂SO₄/HBO₂. Microwave heating resulted in remarkable acceleration of the reaction rate in the first reaction, and had an effect in reducing the reaction time and increasing the yield of rubiadin (4) in the second reaction.  相似文献   

Development of a monoclonal antibody-based sandwich ELISA for detection of guinea pig interleukin-2     

Adachi S  Hashimoto T  Takeyoshi M  Kato H  Iwata H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2006,68(12):1281-1287

Interleukin 2 (IL-2) is a T cell proliferation factor released by Th0- and Th1-type helper T cells and is an essential cytokine for immune responses. In the present study, recombinant glutathione S-transferase (GST)-guinea pig IL-2 (GPIL-2) fusion protein was prepared by Escherichia coli (E. coli) and by using this protein as an immunogen, monoclonal antibodies (mAbs) against GPIL-2 were produced to establish a basis for a research on immune responses in guinea pigs. Three stable hybridoma cell lines were established, and specific binding of each mAb to recombinant GPIL-2 produced by E. coli and insect cells infected with recombinant baculovirus was shown by enzyme linked immunosorbent assay (ELISA) and/or immunoblot analysis. Isotype analyses of these mAbs revealed that all three mAbs were IgG1 and had kappa chain. Furthermore, assessment of their epitopes by competition binding assay indicated that the mAbs obtained in this study bound to three different epitopes. Thus, a sandwich ELISA based on the two mAbs specific to different GPIL-2 epitopes was developed for detection of GPIL-2, which had a sensitivity threshold of about 0.3 ng/ml of GPIL-2.  相似文献   

Lymphography of the thoracic duct by percutaneous injection of iohexol into the popliteal lymph node of dogs: experimental study and clinical application     

Naganobu K  Ohigashi Y  Akiyoshi T  Hagio M  Miyamoto T  Yamaguchi R 《Veterinary surgery : VS》2006,35(4):377-381

OBJECTIVE: To evaluate the efficacy of percutaneous administration of iohexol into the popliteal lymph node as a non-invasive technique for thoracic duct lymphangiography in dogs. STUDY DESIGN: Experimental study and clinical report. ANIMALS: Normal adult dogs (n=4) and 1 dog with recurrent chylothorax. METHODS: For the experimental study, 4 dogs (weight, 8.4-12.3 kg) had 5-10 mL iohexol injected percutaneously into 1 popliteal lymph node and then thoracic radiographs were taken. Popliteal lymph nodes were examined by histopathology 8 days later. One 25-kg dog with recurrent chylothorax had 25 mL iohexol injected into the right popliteal lymph node followed by thoracic radiography. RESULTS: In experimental dogs, the thoracic duct was best visualized on thoracic radiographs after administration of 10 mL iohexol. Clinically, no abnormalities were identified in the injected limb and except for 1 dog that had large numbers of siderocytes and erythrophagocytic macrophages in the injected lymph node, the histopathologic findings in the other injected popliteal lymph nodes were not different from contralateral nodes. In the clinical case, the thoracic duct was visualized, but there was leakage of iohexol around the node. CONCLUSION: The thoracic duct in dogs can be visualized by lymphography after percutaneous injection of iohexol (1 mL/kg at 2 mL/min) into the popliteal lymph node. CLINICAL RELEVANCE: Percutaneous popliteal lymph node administration of iohexol should be considered as an alternative to mesenteric lymph node injection for radiographic identification of the thoracic duct in dogs.  相似文献   

Antioxidant mechanism studies on ferulic acid: identification of oxidative coupling products from methyl ferulate and linoleate     

Masuda T  Yamada K  Maekawa T  Takeda Y  Yamaguchi H 《Journal of agricultural and food chemistry》2006,54(16):6069-6074

In our studies of the chain-breaking antioxidant mechanism of natural phenols in food components, ferulic acid, a phenolic acid widely distributed in edible plants, especially grain, was investigated. The radical oxidation reaction of a large amount of ethyl linoleate in the presence of the methyl ester of ferulic acid produced four types of peroxides as radical termination products. The isolation and structure determination of the peroxides revealed that they had tricyclic structures which consisted of ethyl linoleate, methyl ferulate, and molecular oxygen. Based on the formation pathway of the products, a radical scavenging reaction occurred at the 3'-position of the ferulate radical with the four types of peroxyl radicals of ethyl linoleate. The produced peroxides subsequently underwent intramolecular Diels-Alder reaction to afford stable tricyclic peroxides.  相似文献   

Analyses of leaves from open field-grown transgenic poplars overexpressing xyloglucanase     

Tomomi Kaku  Kei’ichi Baba  Toru Taniguchi  Manabu Kurita  Ken-ichi Konagaya  Katsuaki Ishii  Teiji Kondo  Satoshi Serada  Haruka Iizuka  Rumi Kaida  Teruaki Taji  Yoichi Sakata  Takahisa Hayashi 《Journal of Wood Science》2012,58(4):281-289

The transgenic expression of Aspergillus xyloglucanase cDNA (AaXEG2) with 35S promoter in the leaves of open field-grown poplars was studied. The level of xyloglucan in the transgenic poplars was decreased to 15–16% in the non-fertile soil (forest-field soil) and to 21–22% in the fertile soil (farming-field soil) compared with that of the wild-type poplars. The leaves exhibited a smaller surface area with more rounded teeth than those of the wild-type plants, similar to the sun leaf variety that was grown in the incubation room and subsequently greenhoused. The majority of total veins with water-conducting vascular bundles were shorter in the leaves of the transgenic poplars than those of the wild type. This decrease in vein length may result from a decrease in xyloglucan during leaf development, from which large numbers of proteins were markedly downregulated in the leaves of the transgenic plants via proteomic analysis. It seems likely that the leaves of the transgenic poplars came to relax the edges of their tooth rather than extend their veins as a result of the loosening of the xyloglucan cellulose networks in the leaves.  相似文献   

Prevalence of extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae in food-producing animals     

Hiroi M  Yamazaki F  Harada T  Takahashi N  Iida N  Noda Y  Yagi M  Nishio T  Kanda T  Kawamori F  Sugiyama K  Masuda T  Hara-Kudo Y  Ohashi N 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2012,74(2):189-195

To evaluate the diversity of extended-spectrum β-lactamases (ESBL) genes among food-producing animals, 48 isolates of ESBL-producing Escherichia coli isolates were obtained from rectal samples of broilers, layers, beef cattle and pigs, at the slaughterhouse level. ESBL-carrying E. coli were isolated from 60.0% of individual broiler rectal samples, 5.9% of layers, 12.5% of beef cattle and 3% of pigs. One ESBL-producing Klebsiella pneumoniae was isolated from a broiler. The ESBL-positive E. coli isolates from broilers harbored various ESBL genes: bla (SHV-12), bla(CTX-M-2), bla(CTX-M-14), bla(CTX-M-15) and bla(CTX-M-44). The plasmid DNAs were analyzed by restriction patterns. Homogeneous band patterns were yielded in those of K. pneumoniae and E. coli isolates harboring the bla(CTX-M-2) gene from different farms. No genetic relation between the 2 CTX-M-14 ESBL-producing strains was found by pulsed-field gel electrophoresis, although 2 plasmids in these strains, obtained from different broiler farms, were similar to each other. This study provides evidence that the proliferation of CTX-M-producing E. coli is due to the growth of indigenous CTX-M-producing strains and the possible emergence of strains that acquired CTX-M genes by horizontal transfer in different broiler farms. CTX-M-producing coliforms in broilers should be controlled due to the critical importance of cephalosporins and the zoonotic potential of ESBL-producing bacteria.  相似文献