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71.
Kobayashi S Sakatani M Kobayashi S Okuda K Takahashi M 《The Journal of reproduction and development》2007,53(6):1305-1311
Inhibition of specific gene expression using RNA interference (RNAi) is a valuable tool for functional analysis of a target gene. However, there is little information available concerning RNAi for analysis of gene function in relation to the reproductive physiology of follicular cells in ruminants. Thus, the aim of this study was to evaluate the interfering effect of small interference RNA (siRNA) on expression of cyclooxygenase-2 (Cox-2) mRNA and prostagrandin F(2alpha) (PGF(2alpha)) production in bovine cumulus-granulosa (CG) cells. Bovine CG cells were collected from aspirated follicles and cultured. After reaching confluency, two experiments were conducted. In experiment 1, to investigate the effective concentration of siRNA, 0, 100, 250 and 500 pM of Cox-2 siRNA was introduced into the CG cells, respectively. After 24 h, the amount of Cox-2 mRNA expression was measured by RT-PCR and real-time PCR. In experiment 2, to investigate the time required for effective interference of siRNA and Cox-2 activity, 250 pM siRNA was introduced for 0, 3, 6, 12 and 24 h. After culture, the amount of Cox-2 mRNA expression was measured and the culture medium was collected to determine the PGF(2alpha) concentration by enzyme immunoassay. The Cox-2 mRNA expression was not affected by introduction of 100 pM siRNA into CG cells for 24 h, but 250 and 500 pM Cox-2 siRNA significantly reduced the Cox-2 mRNA expression. Moreover, the significant suppressive effect of 250 pM siRNA was observed 6 h after introduction, and the reduction of mRNA expression by RNAi became more obvious over 12 h. On the other hand, the PGF(2alpha) concentration in the culture medium was not significantly different 12 h after siRNA introduction; however, the PGF(2alpha) concentration 24 h after siRNA introduction was significantly decreased compared with the control at the same time point. These results suggest that gene silencing of Cox-2 with siRNA is capable of analyzing the function and expression of specific genes in bovine CG cells. 相似文献
72.
Yoshimasa SASAKI Taketoshi IWATA Masashi UEMA Kenzo YONEMITSU Shizunobu IGIMI Hiroshi ASAKURA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2022,84(6):743
Chicken is a major source of human campylobacteriosis. Chicken meat originates not only from broilers but also from spent layers; however, few reports have documented the prevalence and antimicrobial resistance of Campylobacter spp. in layers in Japan. Therefore, we investigated the prevalence and antimicrobial susceptibility of Campylobacter spp. in 47 layer farms in Japan. Fecal samples were collected from the youngest and oldest flocks on the farm, and Campylobacter spp. was isolated from 46/47 (97.9%) farms. Among the C. jejuni isolates, the resistance rates to ampicillin, tetracycline, and ciprofloxacin were 29.6%, 22.2%, and 19.8%, respectively. The ciprofloxacin resistance rate (7.3%) in C. jejuni isolated from old flocks was significantly (P<0.01) lower than that in young flocks (32.5%). 相似文献
73.
Yoshi -Nori Nakamura Masashi Ando Manabu Seoka Ken -Ichi Kawasaki Yasuyuki Tsukamasa 《Fisheries Science》2006,72(6):1140-1146
ABSTRACT: Using full-cycle cultured (FC) Pacific bluefin tuna (body length [BL], 42.6–66.4 cm; body weight [BW], 1.66–7.40 kg, n = 15), the changes in chemical compositions and histological structure of the cephalal parts of the dorsal ordinary muscles (DOM) occurring with growth were investigated. A positive correlation ( r = 0.9644, P < 0.05) was observed between BL and BW with growth. The protein, lipid and ash contents of DOM and condition factor did not change with growth. However, the glycogen content of DOM increased ( P < 0.05) from approximately 55 cm (BL) in this study. Using optical microscopic photographs, the various shapes of muscle fibers were observed and it was noted that the muscle fiber diameter increased ( P < 0.05) with growth. Using transmission electron microscopic observation, many glycogen granules were observed in muscle fibers (especially, side of connective tissue) of DOM throughout the growth stage in this study. These results indicate that the glycogen content of DOM of FC Pacific bluefin tuna increases before the lipid with growth. 相似文献
74.
75.
Laddawan Krongpong Kunihiko Futami Takayuki Katagiri Makoto Endo Masashi Maita 《Fisheries Science》2008,74(5):1055-1061
Furazolidone, an antibacterial drug that was once widely used in the livestock industry and aquaculture, is now prohibited
in numerous countries. It is difficult to detect residual furazolidone because it is readily metabolized in animal tissues
but, by using and liquid chromatography coupled with tandem mass spectrometry, its metabolite, 3-amino-2-oxazolidinone (AOZ)
can be detected. Here we describe the validity of an enzyme-linked immunosorbent assay (ELISA) kit to detect AOZ in Japanese
eel Anguilla japonica tissue. ELISA is capable of detecting AOZ at 1.0 μg/kg in an eel sample with excellent accuracy and precision. Our results
show that ELISA is suitable for regulatory purposes and for studying the fate of AOZ residues in eel treated with furazolidone.
To measure the persistence of AOZ in eel tissues, eels (1.4–6.5g) were immersed in tanks containing 2 and 10 mg furazolidone/L
for 3 h, and then maintained in a tank supplying well water for the next 160 days. The half-lives of AOZ, calculated from
the linear terminal part of the excretion curve, were 25.0 days in muscle and 21.6 days in liver from fish exposed to 2 mg/L
furazolidone. In the eels treated with 10 mg/L furazolidone, by contrast, high levels of AOZ were detected in liver and muscle,
but the half-lives of AOZ were similar to those in fish treated with 2 mg/L furazolidone. The half-lives of AOZ in eel tissues
were prolonged by the condition of low water temperature. 相似文献
76.
Expression dynamics of bovine MX genes in the endometrium and placenta during
early to mid pregnancy
Takahiro SHIROZU Keisuke SASAKI Manabu KAWAHARA Yojiro YANAGAWA Masashi NAGANO Nobuhiko YAMAUCHI Masashi TAKAHASHI 《The Journal of reproduction and development》2016,62(1):29-35
MX belongs to a family of type I interferon (IFN)-stimulated genes, and the MX protein has
antiviral activity. MX has at least two isoforms, known as MX1 and
MX2, in mammals. Moreover, bovine MX1 has been found to have alternative
splice variants—namely, MX1-a and MX1B. In ruminants, IFN-τ—a type I IFN—is
temporarily produced from the conceptus before implantation and induces MX expression in the
endometrium. However, the expression dynamics of MX after implantation are not clear. In the
present study, we investigated the expression of MX1-a, MX1B and
MX2 in the endometrium and placenta before and after implantation along with the expression
of IFN-α, type I receptors (IFNAR1 and IFNAR2) and
interferon regulatory factors (IRF3 and IRF9). Pregnant uterine samples were
divided into five groups according to pregnancy days 14–18, 25–40, 50–70, 80–100, and 130–150. Tissue samples
were collected from the intercaruncular endometrium (IC), caruncular endometrium (C) and fetal placenta (P).
Although all the MX expressions were significantly higher in the IC and C at days 14–18,
presumably caused by embryo-secreted IFN-τ stimulation, their expressions were also detectable in the IC, C
and P after implantation. Furthermore, IFN-α expression was significantly higher in the IC.
RT-PCR indicated IFNAR1, IFNAR2, IRF3 and
IRF9 mRNA in all the tissues during pregnancy. These results suggest that all the
MX genes are affected by the type I IFN pathway during pregnancy and are involved in an
immune response to protect the mother and fetus. 相似文献
77.
Shinya Yano Megumi Mori Naozumi Teramoto Makoto Iisaka Natsumi Suzuki Masanari Noto Yasuko Kaimoto Masashi Kakimoto Michio Yamada Eri Shiratsuchi Toshiaki Shimasaki Mitsuhiro Shibata 《Marine drugs》2015,13(1):338-353
Photocrosslinked hydrogels reinforced by microfibrillated cellulose (MFC) were prepared from a methacrylate-functionalized fish elastin polypeptide and MFC dispersed in dimethylsulfoxide (DMSO). First, a water-soluble elastin peptide with a molecular weight of ca. 500 g/mol from the fish bulbus arteriosus was polymerized by N,N′-dicyclohexylcarbodiimide (DCC), a condensation reagent, and then modified with 2-isocyanatoethyl methacrylate (MOI) to yield a photocrosslinkable fish elastin polypeptide. The product was dissolved in DMSO and irradiated with UV light in the presence of a radical photoinitiator. We obtained hydrogels successfully by substitution of DMSO with water. The composite gel with MFC was prepared by UV irradiation of the photocrosslinkable elastin polypeptide mixed with dispersed MFC in DMSO, followed by substitution of DMSO with water. The tensile test of the composite gels revealed that the addition of MFC improved the tensile properties, and the shape of the stress–strain curve of the composite gel became more similar to the typical shape of an elastic material with an increase of MFC content. The rheology measurement showed that the elastic modulus of the composite gel increased with an increase of MFC content. The cell proliferation test on the composite gel showed no toxicity. 相似文献
78.
Machiko Yamada Mayuko Otsubo Yuki Tsutsumi Chiaki Mizota Naoki Iida Kazuma Okamura Masashi Kodama Akira Umehara 《Fisheries Science》2013,79(6):923-934
The genus Skeletonema includes phytoplankton species that are important primary producers in marine food chains. Brackish waters have been reported to be one of the important habitats of some species of Skeletonema. To elucidate the species diversity of Skeletonema in brackish waters, we investigated three Japanese brackish bodies of water: the coastal waters of Toyama Bay, a tidal area of the Chikugo River, and a constructed reservoir in Isahaya Bay. We used molecular analysis based on large subunit rDNA and fine morphological structure to identify species. Skeletonema costatum s.s. (sensu stricto) was isolated at salinities as low as 0.6, but Skeletonema dohrnii, Skeletonema subsalsum, and Skeletonema tropicum were not found at salinities below 11.0. S. costatum s.s. could survive transfer from a medium with a salinity of 15 to a salinity of 2, but S. dohrnii did not survive in the same experiment. Only S. costatum s.s. germinated from the sediment of a reservoir in which the salinity was 0.1–1.4; incubation conditions included temperatures of 10, 15, 20, 25, and 30 °C and salinities of 5 and 30. Skeletonema costatum s.s. was identified as the species most adaptable to low-level salinity variations throughout its lifecycle. 相似文献
79.
Hironori Koga Koji Dohi Reiko Yoshimoto Masashi Mori 《Journal of General Plant Pathology》2008,74(3):246-249
Resistance in the leaf blades of rice plants against a virulent race of the rice blast fungus Magnaporthe oryzae was quantitatively examined using a modified spot inoculation method. Numbers of conidia produced in the lesions were affected
by plant age and paralleled the frequency of resistance infection types, which is indicative of whole-plant-specific resistance
(WPSR), in the inoculated leaf sheaths of the corresponding plants. Exogenous abscisic acid treatment, which suppresses WPSR,
also increased the susceptibility of the leaf blades. These results indicate a correlation between the resistance of the leaf
blades and the WPSR in the leaf sheaths. 相似文献
80.
Fukuma Yasufumi Yamane Akihiko Itoh Tomohiro Tsukamasa Yasuyuki Ando Masashi 《Fisheries Science》2012,78(2):451-461
Fish meat was brought to a supercooled state through slow cooling, and changes in the texture, histology, and protein composition
of the meat were investigated. The groups whose storage temperature were lowered by 1.0°C per day (the 1.0°C group) and 0.5°C
per day (the 0.5°C group) began to freeze in the vicinity of −3.5 and −5.0°C, respectively. The freezing point depended on
the fish species; the lowest freezing point was −8.5°C, for the red sea bream in the 1.0°C group. The breaking strength tended
to decrease more slowly in the 1.0°C group, but the collagen fibers collapsed more rapidly in the 1.0°C group. In SDS electrophoresis,
a slight change in the banding patterns was observed, but the relationship between this observation and changes in histology
and physical properties was unclear. This study shows that it is possible to produce a supercooled state in fish meat, and
demonstrates that supercooling is a potential new storage method that lowers the temperature without generating ice crystals. 相似文献