Origin identification of dried seaweed product “nori” by PCR–RFLP analysis of Pyropia yezoensis in the internal transcribed spacer ITS-1 region     

Ken Touhata  Atsushi Namikoshi  Tamami Suzuki  Jun Iguchi  Nanami Mizusawa  Tatsuro Hara  Shintaro Imamura  Takeshi Yabu  Yumiko Yamashita  Michiaki Yamashita 《Fisheries Science》2013,79(5):865-875

Nucleotide sequences in internal transcribed spacer (ITS)-1 region derived from dried nori products produced in Japan, China, and the Republic of Korea were compared. Thalli contained in the Japanese products were genetically homogenous, and their nucleotide sequences in ITS-1 were identical to those of the reference strains of Pyropia yezoensis f. narawaensis. In Chinese products, the thalli were related to P. yezoensis strain Minomiasakusa. In contrast, the thalli in the Korean products were genetically heterogeneous, and several different P. yezoensis strains and other Pyropia spp. were used for dried nori products. In some thalli produced in both China and Korea, the DNA sequences of the ITS-1 region were identical with that of Japan, suggesting that the cultivar strains might have been transplanted from Japan to China in recent years. The 432-bp-long nucleotide sequences in the ITS-1 region of thalli derived from Japanese origin were cleaved to two restriction fragments at 154 and 278 bp by cleavage of PCR-amplified products using MspI. Conversely, almost all of the corresponding sequences derived from China and Korea were lacking MspI or other restriction patterns, except for nori products from some areas that cultivate a closely related strain to the Japanese cultivar.  相似文献   

Origin identification method by multiple trace elemental analysis of intermuscular bones in grilled eel products produced in Japan, China, and Taiwan     

Jun Iguchi  Yumiko Yamashita  Ken Touhata  Takeshi Yabu  Michiaki Yamashita 《Fisheries Science》2013,79(3):531-536

Trace elemental composition in intermuscular bones of grilled eel fillets was analyzed to discriminate the geographic origin of eel products derived from Japan, China, and Taiwan. The intermuscular bones were decomposed with nitric acid-hydrogen peroxide, and twelve elements (Li, Ti, V, Mn, Ni, Co, Zn, Rb, Sr, Ba, Pb, and U) were analyzed by inductively coupled plasma mass spectrometry. Trace elemental composition including six elements: V, Co, Sr, Ba, Pb, and U, in intermusucular bones were significantly different between Japanese domestic eels and imported eels from China and Taiwan. The average content of lead in Japanese eels was 1/4 to 1/6 that in eels imported from China and Taiwan. The data of elemental content in intermuscular bones were used for linear discriminant analysis, and two discriminant models were constructed. In the Japan–China discriminant model, the discriminant probabilities between Japanese origin and Chinese origin were 82.5 and 93.3 %, respectively. In the Japan–Taiwan discriminant model, the discriminant probabilities between Japanese origin and Taiwanese origin were 87.5 and 87.0 %, respectively. Therefore, trace elemental analysis is effective for country-of-origin identification of eels processed as grilled eel fillets.  相似文献   

Evaluation of QT interval prolongation in dogs with heart failure     

Koyama H  Yoshii H  Yabu H  Kumada H  Fukuda K  Mitani S  Rousselot JF  Hirose H  Uchino T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2004,66(9):1107-1111

Comparison of the QT interval and corrected QT interval values that were calculated by the methods of Bazett (QTc1) and Fridericia (QTc2) were made between dogs with or without cardiac diseases to determine the influence of the QT interval on canine heart failure. Upon comparison of the measured values on ECG between the cardiac disease and non-cardiac disease groups, it was observed that the heart rate(HR) was significantly higher in the cardiac disease group than in the non-cardiac disease group, although the QT interval was similar in the two groups. The QTc1 and QTc2 were significantly longer in the cardiac disease group than in the non-cardiac disease group. With the progression of the New York Heart Association Class, the HR tended to increase. The QTc1 and QTc2 became significantly prolonged with the progression of heart failure. Nevertheless, because Bazett's correction formula is known to overcorrect when the HR is high, it was considered that the QTc1 was actually overcorrected by high HR with the progression of heart failure. The QTc2, on the other hand, was only slightly influenced by HR, suggesting that the prolongation was due to the progression of heart failure. These results suggest that the prolongation of QTc2 in cardiac disease reflects the substantial prolongation of the QT interval without the influence of HR. It is suggested that the QTc2 could be a useful parameter for assessing the degree of heart failure in dogs with cardiac disease.  相似文献   

Evaluating antigen-specific cytotoxicity of CD8+ T cells in fish by granzyme B-like activity     

Toda H  Yabu T  Shiba H  Moritomo T  Nakanishi T 《Veterinary immunology and immunopathology》2011,141(1-2):168-172

Granzyme B plays an important role in granule-mediated apoptosis by CTL. It is a well characterized component of the cytolytic machinery in mammals and a candidate for the evaluation of cytotoxic activity of CTL as an alternative to conventional cytotoxicity assay. In this study, we examined the effects of granzyme inhibitors to assess the characteristics of fish granzymes in terms of substrate specificity and the involvement of granzyme B-like in the cytotoxic response. 3,4-dichloroisocoumarin (DCI), which inhibit the activity of serine protease including all members of the granzyme family, markedly suppressed the cytotoxic activity of CTL. However, CTL-mediated cytotoxicity was significantly but not completely suppressed by the addition of carbobenzyloxy-Ile-Glu-Thr-Asp-fluoromethyl ketone (Z-IETD-FMK) that specifically blocks granzyme B activity. These results suggest that additional serine proteases as well as granzyme B-like are involved in cytotoxicity of CTL in fish. We further compared cytotoxicity with the granzyme B-like hydrolytic activity against fluorogenic substrate acetyl-Ile-Glu-Thr-Asp-4-methylcoumaryl-7-amide (Ac-IETD-MCA) and found that granzyme B-like activity correlated well with the cytotoxicity of CTL in ginbuna crucian carp. Present results suggest that the granzyme activity assays is useful to assess cytotoxic activity of CTL in fish in which genetic information on granzymes and specific tools for cytotoxicity assay are not available because of well conserved catalytic triad residues and substrate binding sites in granzyme B throughout vertebrates.  相似文献   

The phylogeny of Anguillicola crassus and A. globiceps based on partial 18S ribosomal RNA sequences     

Hirose  Yabu  Hirono  Aoki 《Journal of fish diseases》1998,21(4):265-271

The parasitic nematodes Anguillicola crassus and A. globiceps infect the swimbladder of eels. To investigate the relationship of these species at the molecular level, the small subunit ribosomal RNA (18S rRNA) genes of A. crassus and A. globiceps were cloned and their nucleotide sequences determined. The 18S rRNA gene of A. crassus and A. globiceps were found to have similar lengths (886 and 888 base pairs, respectively) and demonstrated extensive similarity in their nucleotide sequences, showing 98.8% homology. However, the sequence homology of the 18S rRNA gene of A. crassus with that of other nematodes was found to be in the range of 72.7–89.0% homology, whilst that of A. globiceps was in the range of 73.1–89.4% homology. A phylogenetic reconstruction showed A. crassus to be closely related to A. globiceps. The analysis of nucleotide sequences of the 18S rRNA gene of fish parasites has proven useful in supporting species status and phylogenetic relationships.  相似文献   

Stress-induced apoptosis by heat shock, UV and γ-ray irradiation in zebrafish embryos detected by increased caspase activity and whole-mount TUNEL staining     

Takeshi Yabu  Setsuko Todoriki  Michiaki Yamashita 《Fisheries Science》2001,67(2):333-340

SUMMARY: Environmental stress-induced apoptosis in zebrafish Danio rerio embryos was characterized by assaying caspase-3-like activity and whole-mount terminal deoxynucleotidyl nick-end labeling (TUNEL) staining. Severe stress conditions, such as heat shock at 39°C for 1 h, ultraviolet light at 10–100 mJ/cm² and γ-ray irradiation at 5–20 Gy induced extensive apoptosis in embryos. Apoptotic cells were observed after the bud and 1-somite stages in normal embryos by TUNEL staining, and after stress treatment many TUNEL-positive cells were found in the enveloping and deep cell layers and the larval fin. The caspase-3-like activity increased severalfold during stress-induced apoptosis in a dose-dependent manner. These findings indicate that apoptotic pathways, mediated by caspase-3-like activity, play a major role in zebrafish embryogenesis under stress conditions.  相似文献   

Selenoneine,total selenium,and total mercury content in the muscle of fishes     

Yamashita  Yumiko  Amlund  Heidi  Suzuki  Tamami  Hara  Tatsuro  Hossain  Mohammed Anwar  Yabu  Takeshi  Touhata  Ken  Yamashita  Michiaki 《Fisheries Science》2011,77(4):679-686

Levels of the selenium-containing imidazole compound selenoneine and overall organic selenium were measured in the muscle of fishes by speciation analysis. The method involves monitoring ⁸²Se levels by liquid chromatography inductively coupled plasma mass spectroscopy using a gel filtration column. Selenoneine levels were found to be highest in swordfish muscle (concentration 2.8 nmol/g tissue). The selenoneine contents of bigeye tuna, Pacific bluefin tuna, albacore, yellowfin tuna, and alfonsino muscle were 1.3–2.6 nmol/g tissue. In muscle of these fishes, most organic selenium (9–42%) was present as selenoneine. In other fish species, such as Pacific sardine, greeneye, skipjack, Pacific mackerel, horse mackerel, red sea bream, and Japanese barracuda, selenoneine levels were 0.1–1.4 nmol/g tissue, accounting for 3–34% of organic selenium. In contrast, muscle of Japanese conger, Japanese anchovy, chum salmon, Pacific saury, white croaker, and marbled sole contained levels of selenoneine below the level of detection (<0.05 nmol/g tissue). Mercury and selenium contents were 0.01–5.12 nmol/g tissue and 1.4–19.1 nmol/g tissue. The Se-to-Hg molar ratio varied from species to species, ranging from 1 for swordfish to 217 for marbled sole.  相似文献   

Differential kinetic activities of glycerol kinase among African trypanosome species: phylogenetic and therapeutic implications     

Ohashi-Suzuki M  Yabu Y  Ohshima S  Nakamura K  Kido Y  Sakamoto K  Kita K  Ohta N  Suzuki T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(5):615-621

African trypanosome species are causative agents for sleeping sickness in humans and nagana disease in cattle. Trypanosoma brucei can generate ATP via a reverse reaction with glycerol kinase (GK) when alternative oxidase (AOX) is inhibited; thus, GK is considered to be a crucial target for chemotherapy combined with AOX. However, the energy metabolism systems of African trypanosome species other than T. brucei are poorly understood. Thus, GK genes were surveyed from genome databases and cloned by PCR from T. vivax and T. congolense. Then, recombinant GK proteins (rGK) of T. vivax, T. congolense and T. brucei were expressed and purified. Kinetic analysis of these rGK proteins revealed that the K(m) values of T. congolense rGK for ADP and G-3-P substrates were lower than those of T. vivax and T. brucei. The expression level of GK molecules was highest in T. congolense cells and lowest in T. vivax cells. Based on these results, effective combination dosages of ascofuranone, a specific inhibitor of AOX, and glycerol, an inhibitor of the GK reverse reaction, were determined by using in vitro-cultured trypanosome cells.  相似文献   

Purification and characterization of glutathione peroxidase 1 in the red muscle of Pacific bluefin tuna Thunnus orientalis     

Yamashita  Yumiko  Yabu  Takeshi  Touhata  Ken  Yamashita  Michiaki 《Fisheries Science》2012,78(2):407-413

Glutathione peroxidase GPx1 was purified from the red muscle of Pacific bluefin tuna Thunnus orientalis and its enzymatic properties were characterized. The enzyme was optimally active at pH 7.4 with a specific activity for hydrogen peroxide and a K _m of 6.7 μM. cDNA was also isolated and it contained a predicted open reading frame (ORF) encoding a 188 amino acid protein. The phylogenetic tree shows that fish including Pacific bluefin tuna, pufferfish, and zebrafish, but not mammals, possess two genetically distinct types of GPx1, i.e., GPx1a and GPx1b. The purified enzyme was classified as a fish GPx-1b enzyme on the basis of the phylogenetic tree of the GPx1 family.  相似文献