Gene expression profile of HIRRV G and N protein gene vaccinated Japanese flounder, Paralichthys olivaceus during HIRRV infection

The glycoprotein (G protein) gene, but not the nucleocapsid protein (N protein) gene, of the hirame rhabdovirus (HIRRV) was previously shown to be highly effective in inducing a protective immune response in Japanese flounder (Paralichthys olivaceus) when used as a DNA vaccine. Our previous cDNA microarray analysis demonstrated that interferon-stimulated genes (ISGs) were strongly induced by the HIRRV G protein gene (pHRV-G) but not by the N protein gene (pHRV-N). However, the molecular basis for the difference in protective immunity between pHRV-G- and pHRV-N-vaccinated fish during HIRRV infection is still unclear. In this study, we use a DNA microarray to analyze differences of gene expression in pHRV-G- and pHRV-N-vaccinated fish during HIRRV infection. Microarray analyses showed substantial difference in gene expression patterns during HIRRV infection between fish vaccinated with pHRV-G and pHRV-N. In addition, genes having homology to mammalian T cell activation-related genes were up-regulated in the HIRRV G protein-vaccinated group.     

Generation of monoclonal antibodies specific for ORF68 of koi herpesvirus

Outbreaks of koi herpesvirus (KHV) infection in carp are still a serious problem worldwide. KHV is closely related to other two cyprinid herpesviruses, pox herpesvirus (CHV) and haematopoietic necrosis herpesvirus (CyHV-2) in goldfish. In this study, two major KHV antigenic proteins (ORF62 and ORF68) were identified by immunoscreening using a KHV-specific polyclonal antibody, and then monoclonal antibodies were generated for immunodiagnostic studies. After screening hybridoma cells, one mAb against ORF68 (mAb-7C6) was obtained but no mAbs against ORF62. mAb-7C6 specifically reacted with a lysate of KHV-infected koi fin cells (KF-1 cells) but not with lysates of CHV- or CyHV-2-infected KF-1 cells in an immuno-blotting analysis. Similar results were shown in the following tests: (1) a indirect fluorescent antibody test using infected KF-1 cells and (2) an immunohistochemical investigation by fast red stain (infected liver) or FITC detection (infected spleen). These results suggested that mAb-7C6 specifically reacts with KHV ORF68 protein.     

Genotypic diversity of Theileria orientalis detected from cattle grazing in Kumamoto and Okinawa prefectures of Japan

Theileria orientalis is a benign protozoan species that is widely distributed in Japan, yet sometimes causes serious economic losses in the livestock industry. In this study, we conducted a molecular survey based on genes encoding the major piroplasm surface protein (MPSP) and p23 for T. orientalis detected in cattle grazing in southern areas of Japan, consisting of 2 farms in Kumamoto prefecture (Aso and Kuma districts) and 3 farms in Okinawa prefecture (Ishigaki, Iriomote, and Yonaguni Islands). High prevalence rates of T. orientalis infection were shown in all the cattle populations using the diagnostic MPSP- and p23-PCR assays. Phylogenetic analyses revealed 4 MPSP genotypes and 3 p23 genotypes. Furthermore, MPSP genotype-specific PCR methods were developed in this study and wide distributions of 5-district genotypes of T. orientalis were observed for the examined farms. Our results indicate that at least 5 types of T. orientalis exist in Kumamoto and Okinawa prefectures of Japan and that genotype-specific PCR assays are highly applicable for the quarantine of transported cattle and for epidemiological surveys of bovine theileriosis in Japan.     

Single-copy nuclear PolA1 gene sheds light on the origin of S genome with relationships to B and G genomes of polyploid wheat species

PolA1, a single-copy nuclear gene encoding the largest subunit of RNA polymerase I, comprises highly polymorphic intron 19 and nucleotide tag (Ntag) sequences. We analyzed these sequences in 42 accessions, which differed in ploidy, of Triticum–Aegilops and Hordeum species. The lengths of the intron 19 sequences were ca. 110?bp long in Triticum–Aegilops species, except in four Sitopsis species, Ae. longissima, Ae. searsii, Ae. sharonensis, Ae. speltoides, which had introns similar in length to those of Hordeum species, i.e., ca. 240?bp long. Phylogenetic analyses of the Ntag sequences showed that the four Sitopsis and remaining Triticum–Aegilops species were classified into two discrete Hordeum and Triticum clades, respectively. The A and D genome-specific Ntag sequences of polyploid wheats were highly homologous with those of T. urartu and Ae. tauschii, respectively. In Ae. bicornis, another Sitopsis species, two accessions had the short intron 19 and Triticum–type Ntag sequence, which were highly homologous with those of the B genome in polyploid wheats, whereas one accession contained the long intron 19 and Hordeum–type Ntag sequences. In contrast, partial sequence analyses revealed that the three accessions of Ae. bicornis shared highly homology to single-copy DMC1 and EF-G genes. The discrepancy between these results indicates that the Sitopsis species were probably established by hybrid speciation including ancient introgressive hybridization between progenitors of Triticum–Aegilops and Hordeum. Although many researchers have proposed Ae. speltoides as a candidate for the B genome donor, our data suggest the existence of diploid B genome species in the past that were responsible for the origin of both polyploid wheats and Sitopsis species, including Ae. speltoides.     

Presence of a Temperature Gradient Among Genital Tract Portions and the Thermal Changes Within These Portions Over the Estrous Cycle in Beef Cows

The aim of the present study was to describe the temperature of the different portions of the female genital tract and their relation to rectal temperature and to investigate the effect of steroid hormones profiles on these variables over the estrous cycle in cattle. Four nonpregnant Japanese Black cows were investigated daily over two successive estrous cycles using a digital thermometer with a long probe and rounded-end sensor to record the temperature of the rectum (RT), vagina (VT), cervix (CT), uterine body (UBT) and uterine horns (UHT). Blood samples were collected immediately before temperature recording to assay peripheral levels of progesterone (P₄) and estradiol-17β (E₂). Moreover, transrectal ultrasonography was carried out after temperature recording to monitor the ovulatory follicle and track ovulation. During the experiment, the ambient temperature and relative humidity were recorded for further calculation of the temperature humidity index (THI). The temperature within the genital tracts in these cows progressively increased towards the uterine horns from the vagina. The VT, CT, UBT and UHTs were significantly higher in association with peripheral P₄ concentrations greater than 4 ng/ml (mid-luteal phase) when compared with lower peripheral P₄ concentrations. The VT was more significantly (P<0.01) correlated to the CT, UBT and UHTs than RT. In conclusion, a temperature gradient was present among the vagina, cervix and uterus over the estrous cycle, and changes in peripheral P₄ concentrations were associated with the thermal variations within these portions. The VT could be more beneficial than RT in monitoring temperature of deeper portions of the female genital tract in bovine.     

Cloning of estrogen receptor-β cDNA of common Japanese conger Conger myriaster and expression of the gene during oocyte development

ABSTRACT:   Estrogen receptor (ER) cDNA was cloned from the liver of common Japanese conger Conger myriaster . Common Japanese conger ER cDNA contains 2543 nucleotides including an open reading frame that encodes 596 amino acids, with a molecular weight of 66.1 kDa. The deduced amino-acid sequence of the common Japanese conger ER shares 85% identity with that of eel Anguilla japonica ER-β, and 39–46% and 53–59% identity with the deduced sequences of ER-α and ER-β of other species, respectively. In a phylogenetic analysis, the common Japanese conger ER was found to cluster with the ER-β of other species. The common Japanese conger ER-β also has six domains (A–F), which are conserved in the nuclear receptor superfamily. The amino-acid sequences in the C and E domains were highly conserved between common Japanese conger ER-β and other ER. However, the other domains showed low homology. In the female common Japanese conger, ER-β mRNA was highly expressed in the liver and pituitary gland. The expression levels of the ER-β gene increased from the oil droplet stage to the primary yolk globule stage and were maintained until the migratory nucleus stage.     

Preventive effects of amino acids and glutathione on the formaldehyde-induced denaturation of myofibrillar proteins

ABSTRACT:     Formaldehyde (FA)-induced denaturation of myofibrillar proteins and its prevention were investigated by means of measuring the solubility, adenosine triphosphatase (ATPase) activity, and thermal gel formability of myofibrils and surimi proteins in the presence and absence of free amino acids and glutathione, reduced form. The addition of FA decreased the solubility of myofibrils in 0.5 M NaCl at pH 7.0 and 0°C depending on its concentration and incubation time. The solubility decrease was completely inhibited by the presence of equal, twofold, and threefold amounts of cysteine (Cys), glutathione, and histidine (His) to the amount of FA, respectively. Myofibrillar Ca-ATPase was markedly activated at the initial phase and then decreased later by the addition of FA. The K-ATPase was inactivated with an increase in the amount of FA. The FA-induced changes in both ATPase activities were inhibited in the presence of Cys and His. Thermal gel formability of surimi paste increased only in a short period after the addition of a low concentration of FA. Practically, FA inhibited the thermal gelation and setting effect through the inactivation of transglutaminase. In the presence of Cys, His or glutathione, a strong elastic surimi gel was produced because FA-induced detrimental effects were inhibited.     

毛乌素沙地杨树年轮结构对气候因子的响应

该文应用年轮气候学理论和方法对毛乌素沙地小叶杨和北京杨年轮结构的变化特征及其与月平均气温和月降水量的响应关系进行了分析.结果表明，小叶杨和北京杨的年轮宽度指数序列的变动模式很相似，但小叶杨年轮宽度指数的变动幅度比北京杨大得多，这与毛乌素沙地的气候变化大的特点相一致.用月平均气温和月降水量作为气候因子建立的响应函数模型，能够说明2种杨树年轮宽度和导管腔径变化的46%～55％.小叶杨的年轮宽度和导管腔径对月平均气温及月降水量的反应基本上是一致的.当年2月气温过高会抑制小叶杨年轮宽度和导管腔径的增加，而夏季降水量和气温对其年轮宽度和导管腔径增大有明显的促进作用.北京杨的年轮宽度和导管腔径对月平均气温及月降水量的反应不一致，并且对气候因子的响应关系不明显.