Age and growth of Sebastes vulpes in the coastal waters of western Hokkaido, Japan

ABSTRACT:   Age and growth of Sebastes vulpes collected from the coastal waters of western Hokkaido were studied. Observation of the otolith margin verified that annuli (outer margins of the opaque zone) were produced chiefly from July to August. This period was associated with parturition and the birth season. The maximum age estimated by the surface method was 12 years but the oldest fish was aged at 35 years by the cross-section method. The surface method was inadequate for aging of S. vulpes older than 6 years because of the underestimation of age. No significant difference was found in the parameters of the growth equations between both sexes. The von Bertalanffy growth curve combined for both sexes was as follows: SL _t  = 358.6(1 − exp^{−0.156( t +0.820)}), where SL _t is standard length (mm) at age t (after parturition in years). It seems likely that S. vulpes grows slowly and lives longer than previously thought.     

First record of avian Plasmodium DNA from mosquitoes collected in the Yaeyama Archipelago, southwestern border of Japan

We studied the prevalence of avian Plasmodium in 509 mosquitoes of 9 species collected from the Ishigaki and Iriomote islands in the Yaeyama Archipelago, located southwest from the mainland of Japan. Two identical avian Plasmodium lineages were detected from Culex (Culiciomyia) nigropunctatus. Detected lineages were phylogenetically classified into different clade to avian Plasmodium lineages from birds and mosquitoes in the mainland of Japan but identical to a lineage detected from a resident bird, White-breasted Waterken (Amaurornis phoenicurus). This is the first detection of avian Plasmodium DNA from mosquitoes in the Yaeyama Archipelago and suggested that resident birds might have been infected with an avian Plasmodium lineage specific to the studied area and C. nigropunctatus could be the candidate vector mosquito species.     

The biological activities of cardenolide triglycosides from stems, twigs, and leaves of Nerium oleander

Sixteen cardenolide triglycosides (1?C16) were isolated from stems, twigs, and leaves of Nerium oleander. Among them, 3??-O-(4-O-gentiobiosyl-d-diginosyl)-7??,8-epoxy-14-hydroxy-5??,14??-card-20(22)-enolide, named cardenolide B-3 (16), was isolated from natural sources for the first time. The in vitro anti-inflammatory activities of compounds 1?C16 were examined on the basis of inhibitory activity against the induction of intercellular adhesion molecule-1 (ICAM-1). Compounds 1?C5 were active at an IC50 value of less than 7 ??M. The cytotoxic activity of isolated compounds was evaluated against three human cell lines: normal human fibroblast cells (W-38), malignant tumor cells induced from WI-38 (VA-13), and human liver tumor cells (HepG2). Compounds 1?C5 were active toward WI-38 cells; compounds 1, 3, and 5 were active toward VA-13 cells; and compounds 1?C5 were active toward HepG2 cells at IC₅₀ values of less than 10 ??M. The multidrug-resistant (MDR) cancer-reversal activity of compounds 1?C16 was evaluated on the basis of the amount of calcein accumulated in MDR human ovarian cancer 2780AD cells in the presence of each compound. Compounds 13 and 14 showed significant effects on calcein accumulation.     

Mobilization of the active transposon mPing in interspecific hybrid rice between Oryza sativa and O. glaberrima

Miniature Ping (mPing) is the first active miniature inverted-repeat transposable element to be identified in rice, and its mobilization is activated by stress treatments. We have examined the mobilization of mPing in four NERICA (New Rice for Africa) lines and 13 interspecific lines. All 17 lines are inbred progenies derived from crosses between Oryza sativa variety WAB56-104 as the recurrent parent and the O. glaberrima variety CG14 as the donor parent. We found that 16 of the 17 lines studied inherited mPing together with its autonomous partner, Pong, from WAB56-104. Transposon display of mPing disclosed polymorphic banding patterns among these lines. Most importantly, seven of the lines displayed clear polymorphic banding patterns for mPing, indicating that mPing might have been mobilized in these lines. Locus-specific PCR analysis also confirmed the mobilization of mPing. These results signify that interspecific hybridization may activate the transposition of mPing. Based on these results, we discuss the potential use of the mPing system as an efficient tool for gene tagging in interspecific hybrid rice.     

Construction of an expression vector containing a β-actin promoter region for gene transfer by microinjection in red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

ABSTRACT:   Transgenic technology has been widely applied to a variety of freshwater fish species. However there are few reports on the use of this technology in commercially important marine species. In this study, the construction of expression vectors containing the β-actin promoter region for use in the red sea bream Pagrus major , a species of considerable importance to the aquaculture industry in Japan is reported. The β-actin gene was cloned from a red sea bream genomic DNA library. Recombinant plasmids were constructed by linking the 5' flanking region of the β-actin gene to the green fluorescent protein reporter gene, followed by the poly A signal sequence of simian virus 40 or the 3' flanking region the β-actin gene. Expression of these constructs was examined following microinjection into zebrafish and red sea bream embryos, and compared to that of the expression vector pXI-GFP driven by the Xenopus elongation factor 1α. The results indicated that the construct consisting of the β-actin 5'-and 3' flanking regions was the most efficacious. In future studies, it is planned to investigate the efficient condition for integration into chromosomes of the transgene.     

First report of tomato chlorotic dwarf viroid isolated from symptomless petunia plants (Petunia spp.) in Japan

A viroid was detected for the first time in symptomless petunia plants (Petunia spp.) and identified as Tomato chlorotic dwarf viroid (TCDVd) based on an analysis of the complete genomic sequence. These petunia plants are a likely source of inoculum for tomato or potato plants because TCDVd induces severe symptoms on these plants. The genomic sequence of this petunia isolate from Japan shared 100 % identity with petunia isolates from the Netherlands and United Kingdom and a tomato isolate from Japan. Phylogenetic analysis showed that all petunia isolates and the tomato isolate from Japan formed a monophyletic clade.     

The development of a sensitive enzyme immunoassay for the determination of estrone and estradiol-17beta in bovine blood plasma based on the same homologous combination with antiserum and steroid-enzyme conjugate

The development of a sensitive enzymeimmunoassay (EIA) for the determination of estrone (E1) and estradiol-17beta (E2 beta) in bovine plasma is described. The assay is a homologous double-antibody EIA with E2beta 17hemisuccinate (HS) as hapten for the immunoreactive reagent. The antiserum was raised against E2beta 17HS bovine serum albumin conjugate in the rabbit, and E2beta 17HS-horseradish peroxidase was used as steroid-enzyme conjugate. Each estrogen EIA was distinguished only by using the each working standard and sample for the EIA. Bovine plasma E1 and E2beta were extracted and purified before EIA. The antiserum was used at 1:1,750,000 dilutions for EIA. Estrone and E2beta showed high cross-reactivity with the antiserum (E1: 350.7%, E2beta:100%). The sensitivities were <0.03 pg/well for E1 and <0.12 pg/well for E2beta. Recovery rates of E1 and E2beta added to bovine blood plasma were 94.5% and 93.9%, respectively. The precision for EIA of estrogens was below 9.7%. The profiles of either estrogen as determined by EIA corresponded closely well with follicle dynamics in the cow during the estrous cycles and with placental function in pregnant animals. In conclusion, our new EIA can be applied with sufficient sensitivities, recovery and precision for the routine analysis of E1 and E2beta concentrations in bovine plasma.     

Effect of germanium,poly-trans-[2-carboxyethyl] germasesquioxane on natural killer (NK) activity in dogs

In an attempt to study the immunological effects on normal and adenocarcinoma (AC) dogs, the natural killer (NK) activity was determined. Augmentation of NK cell activities in responsive normal and AC dogs that indicatedanergy in the phaseolus vulgaris agglutinin (PHA) skin-test was manifested when the animals were accordingly treated with germanium (GN).     

Application of DNA markers for discrimination between Japanese and Australian Wagyu beef

The objective of this study was to discriminate between original Japanese and Australian Wagyu beef, which is sold in the Singapore markets, using six previously developed DNA markers. To effectively evaluate the six markers for breed identification, the probability of identification as Australian Wagyu beef was calculated based on the estimated allele frequencies using 130 Australian Wagyu individuals. The combined use of six markers would allow the discrimination of Australian Wagyu beef with an estimated probability of 0.776. The probability to discriminate Australian Wagyu from Japanese Wagyu beef was sufficiently high. In addition, Australian Wagyu has maternal mitochondrial DNA of Bos indicus cattle with moderate high frequency of 0.377. The DNA marker system could also be used as a deterrent force against false sales, and contribute to the reduction and prevention of incorrect or falsified labeling of beef.     

Polymorphisms of the growth hormone gene in domesticated red sea bream populations (Pagrus major) based on minisatellite genotypes and nucleotide sequences

The genetic diversity of the growth hormone gene in domesticated red sea bream (pmaGH) was evaluated using a minisatellite DNA marker located in intron 3 (pmaGH22) and nucleotide sequences. The number of alleles of pmaGH22 was largely decreased in domesticated strains of red sea bream, and the possibility of selection pressures was also detected based on the analysis of the Hardy–Weinberg equilibrium in some strains. However, each strain inherited a small number of alleles of pmaGH22, and the entire domesticated population (combining all strains) showed a large number of alleles (n = 17), similar to the allelic richness of the wild population (n = 18.5). Based on nucleotide sequencing analysis, three synonym mutations were found in the coding regions, and also several SNPs and indels were found in the noncoding regions. In addition, four genealogies of growth hormone haplotypes were identified based on principal coordinate analysis, and these genealogies of pmaGH partly reflected allele size ranges of pmaGH22. Several haplotypes shared alleles of pmaGH22, and also fragment size homoplasy in pmaGH22 was suspected. These alleles of pmaGH22 and the haplotypes will be a useful indicator for divergence of pmaGH and for broodstock individual selection with minimum inbreeding effect.