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1.
H Inokuma G Nane T Uechi Y Yonahara P Brouqui M Okuda T Onishi 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2001,63(11):1225-1227
Twelve (54.5%) of 22 free-roaming dogs in Ishigaki Island had tick infestation identified as Rhipicephalus sanguineus. There were 121 ticks recovered and consisted of 28 females, 58 males, 22 nymphs and 3 larvae. Infection of dogs possibly with canine ehrlichial pathogens was examined by both indirect immunofluorescence assay and polymerase chain reaction (PCR). Two dogs of the 13 examined were sero-positive for the human granulocytic ehrlichia agent, and one of two dogs was PCR positive for Ehrlichia platys. This dog had platelet numbers slightly lower than normal value, however, no morulae were found within platelet on peripheral blood smear stained with Giemsa. 相似文献
2.
Wei-Qin Wang Tomohide Natsuaki Yoshitaka Kosaka Seiichi Okuda 《Journal of General Plant Pathology》2006,72(1):52-56
To identify possible sites of viral attenuation, the complete nucleotide sequences of two isolates of Zucchini yellow mosaic virus (ZYMV) were determined; a severe isolate Z5-1 and an attenuated isolate from Z5-1 (designated ZYMV-2002). The viral genome
of both isolates consisted of 9593 nucleotides in size and contained an open reading frame encoding a single polyprotein of
3080 amino acids. Comparison of the nucleotide sequences for Z5-1 and ZYMV-2002 revealed 14 nucleotide mutations, resulting
in seven amino acid substitutions with four in the HC-Pro region, two in the CI region, and one in the NIb region. These results
provide a genetic basis for future manipulation of the ZYMV reverse genetics system.
The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB188115
and AB188116 相似文献
3.
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
4.
It is believed that some organic compounds form complexes with iron and aluminum and prevent the fixation of phosphate applied to soils. 相似文献
5.
Tomoko ITAHASHI Toshinori OIKAWA Takashi NUMABE 《The Journal of reproduction and development》2022,68(1):74
This study was conducted to examine the effects of adding glutathione (1 mM) to media used for sperm washing and in vitro fertilization (IVF) on the improvement of early development of embryos produced using cryopreserved spermatozoa of the less IVF-competent bull (the one considered unqualified as spermatozoa supplier for the production of bovine blastocysts using IVF). The cryopreserved spermatozoa of this bull were characterized by normal motility and lower ATP content and blastocyst productivity than those of IVF-competent bulls. The addition of glutathione to the sperm washing medium was more effective in improving the productivity of blastocysts and ATP content than the addition of glutathione to the IVF medium or no glutathione addition at all (control). These results suggest that this simple method may be used to improve the potential of cryopreserved spermatozoa of less IVF-competent bulls to fertilize oocytes in vitro and to induce normal embryonic development after fertilization. 相似文献
6.
Anzai T Eguchi M Sekizaki T Kamada M Yamamoto K Okuda T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1999,61(12):1287-1292
In order to establish a rapid diagnostic method for contagious equine metritis (CEM), we developed and evaluated a polymerase chain reaction (PCR) test. Species-specific PCR primer sets were derived from the DNA sequence of a cloned DNA fragment of Taylorella equigenitalis that did not hybridize with the genome of a taxomonically related species, Oligella urethralis. Single step PCR with primer set P1-N2 and two-step semi-nested PCR with primer sets P1-N2 and P2-N2 detected as low as 100 and 10 CFU of the bacteria, respectively. Single-step PCR detected T. equigenitalis from genital swabs of experimentally infected mares with sensitivity comparable to that of bacterial isolation. Furthermore, two-step PCR was more sensitive than the culture method. Upon examination of field samples, 12 out of 3,123 samples were positive by single-step PCR while only 2 were positive by bacterial culture. The 12 PCR-positive samples originated from 5 mares, of which 3 animals were considered to be carriers based on previous bacteriologic and serologic diagnoses for CEM. The PCR test described in this study would provide a specific and highly sensitive tool for the rapid diagnosis of CEM. 相似文献
7.
Nakano S Kishi H Ogawa H Yasue H Okano A Okuda K 《The Journal of reproduction and development》2003,49(2):127-134
We investigated endometrial expression of trophinin mRNA and protein, homophilic cell adhesion molecules, during the estrous cycle of gilts. An immunopositive reaction for trophinin was observed in the luminal and glandular epithelia of the endometrium at all stages of the estrous cycle, but not in endometrial stromal cells or the myometrium. A partial coding sequence of porcine trophinin was similar to sequences in humans and mice, with homologies of 75% and 70%, respectively. As in humans and mice, the trophinin gene is expressed in the endometrium. Trophinin, however, is expressed in the endometrium of the pig throughout the estrous cycle, higher expression levels were observed at some points of the luteal phase, as in humans. These findings suggest that regulation of trophinin gene expression in the pig is different from that in mice, but similar to that in humans. Furthermore, the present results suggest that the pig might be a suitable model for studying the physiological importance of trophinin in early pregnancy in humans. 相似文献
8.
Shibata I Okuda Y Yazawa S Ono M Sasaki T Itagaki M Nakajima N Okabe Y Hidejima I 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(3):405-408
Porcine circovirus type 2 (PCV2) shedding patterns were investigated by polymerase chain reaction (PCR) for the detection of PCV2 DNA, and the diagnostic suitability of a sample for the PCR was examined by using different types of samples. In the experimental infection, sixteen pigs were inoculated intranasally with PCV2. The samples, including oropharyngeal and nasal swabs, feces, whole blood and serum became positive for PCV2 DNA by PCR immediately after the inoculation, and almost all samples remained positive during the observation period, post-inoculation-day 70. Field samples were collected from 313 pigs in five different age groups. The overall percentages of positive samples in the whole blood, nasal swabs, and feces detected by PCR were 30.4%, 19.2%, and 20.4%, respectively. The frequency of positive samples increased after the nursery stages and reached a peak in the 3 to 4-month-old pigs. These results indicate that PCV2 infection may occur after weaning, that PCV2 DNA may be present in whole blood for a long period after infection, and that whole blood and serum are the most suitable sample types for the PCR analysis of PCV2. 相似文献
9.
Matsumoto K Inokuma H Okuda M Onishi T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(1):137-140
The effects of salivary gland extract (SGE) from R. sanguineus were examined on the production of IgG1 and IgG2 and the mRNA expression of IFN-gamma, IL-2, IL-4, IL-5 and IL-10 in the mononuclear cells from canine peripheral blood, treated with concanavalin A (ConA) in vitro. SGE suppressed the ConA-induced production of IgG2. It also inhibited the expression of IFN-gamma, IL-2 and IL-5 mRNA in a dose-dependent manner. No dose-dependent suppression was observed of IL-10 mRNA expression although a significant effect was observed at a SGE protein concentration of 25 microg/ml. SGE had no effect on the mRNA expression of IL-4. These results suggest that the suppression of IgG2 production by SGE from R. sanguineus was caused by the suppression of IFN-gamma production. 相似文献
10.