Changes in Serum Steroid Hormones and Vitellogenin Levels in Cultured Female European Eels Anguilla anguilla during Artificially Induced Ovarian Development

Changes in serum concentrations of estradiol-17β, testosterone, 17α,20βdihydroxy-4-pregnen-3-one, and vitellogenin were investigated during ovarian development induced by injection of a salmon pituitary extract in cultured European eel Anguilla anguilla . Vitellogenesis was induced with a weekly dose of 50 mg pituitary extract/kg body weight. In eels receiving that dose, gonadsomatic indices ranged from 20–43% after the 10th–11th weekly injection. Body weights were relatively stable during vitellogenesis, but increased dramatically during final maturation. Serum estradiol-17β levels increased slightly during vitellogenesis and peaked at an average of 6.95 ng/mL at final maturation. The profile of serum vitellogenin followed that of estradiol-17β which increased markedly from an average of 0.36 to 20.72 mg/mL. Control levels of vitellogenin were undetectable throughout the study. Average serum levels of testosterone increased to a peak of 17.74 ng/mL in the early stage of vitellogenesis, followed by a sharp drop to initial levels (3.86 ng/mL) in the late stage of vitellogenesis, and then increased again to an average of 8.84 ng/mL at the final maturation stage. Serum 17α,20β-dihydroxy-4-pregnen-3-one was not detected (<50 pg/mL) throughout the experiment. Profiles of serum estradiol-17β, testosterone, and vitellogenin observed during ovarian development appear different from those found in salmonids and other teleosts. This study indicates, however, that cultured European eels are a useful model for study of gonadal maturation in the eel.     

Ultrastructural study on interaction between a sterile,white endophytic fungus and eggplant roots

Eggplant roots colonized by a sterile, white mycelial endophyte (SWM) were previously found to become highly resistant to Verticillium wilt. SWM alone, however, caused no visible, disease symptoms, such as wilting or necrosis. The mechanism of the symptomless infection by SWM was investigated in this study. Electron microscopy revealed that hyphae of SWM were abundant on and inside the root epidermal cells 2 weeks after inoculation. Many terminal appressoria formed from apical tips of hyphae, and heavy degradation of the host cell walls was evident where hyphae accumulated. By 4 weeks following inoculation, penetration pegs easily breached epidermal cells, and the infection hyphae penetrated outer cortical cells. In response to the hyphal ingress, numerous tubule-like vesicles and membrane-bound, multivesicular bodies accumulated in cortical cytoplasm near the infection sites of the outer cortical cells, but no visible signs of the host reactions were seen in the epidermal cells. Papillae developed at the spaces between cell walls and plasma membranes at the infection sites. The penetration hyphae often grew out of the papillae, but further hyphal ingress was halted in the middle cortical cell layer. By 8 weeks following inoculation, papillae that developed in these cells contained larger amounts of highly electron-dense material and were reinforced by multilamellate, fibrous elements. Hyphae that entered such papillae were confined to them, and the hyphal cytoplasm degenerated. As the result of the activated resistance reactions, root vascular cylinders remained intact, and the host plants did not wilt.     

Effect of placement of urea and coated urea fertilizers on yield and quality of soybean (Glycine max (L.) Merr.) seeds

The objective of the present study was to record the seed yield and to examine visually the quality of soybean seeds cultivated under different types and placements of urea fertilizers. In addition to the conventional fertilizer application (including ammonium sulfate 16 kg N ha^-1 broadcasting (100 kg N ha^-1 of urea (0B) and X00-d type coated urea CU-100 (CUB), and deep placement (100 kg N ha^-1) of urea (UD) and 100-d type coated urea CU-100 (CUD) was conducted in separate plots in a paddy field converted to an upland field located at Shindori Experimental Station of Niigata University. Soybean plant growth was periodically analyzed and the quality of harvested seeds was also visually examined (hereafter referred to as “visual quality”). It was found that the deep placement treatments were more conducive 1o nitrogen (N₂)fixation, based on the relative mreide N concentration in the xylem sap, which is a good indicator of N~fixation by soybean. Also the total seed yield was the highest in CUD (82 g plant^-1) and 0D (81 g plant^-1), compared to the control (62 g plant^-1), UB (68 g plant^-1), and CUB (68 g plant^-1). The visual quality of harvested seeds showed that CUD enhanced the quality of seeds compared to the other treatments, in which the percentage of good quality seeds, hereafter referred to as "good seeds," based on the dry weight was 51 (control), 65 (K3B), 61 (CUB), 61 (0D), and 6696 (CUD). In terms of diseased seeds, the percentage of turtle wrinkle and broken seed coats was found to decrease by N application compared to the control. Thus, it is suggested that N fertilization management is important for maximum yield of soybean as well as for the enhancement of seed quality.     

Gene expression and hormonal regulation of adiponectin and its receptors in bovine mammary gland and mammary epithelial cells

Although the functions of adiponectin, a differentiated adipocyte‐derived hormone, in regulating glucose and fatty acid metabolism are regulated by two subtypes of adiponectin receptors (AdipoRs; AdipoR1 and AdipoR2), those in ruminants remain unclear. Therefore we examined the messenger RNA (mRNA) expression levels of adiponectin and its receptors in various bovine tissues and mammary glands among different lactation stages, and the effects of lactogenic hormones (insulin, dexamethasone and prolactin) and growth hormone (GH) on mRNA expression of the AdipoRs in cultured bovine mammary epithelial cells (BMEC). AdipoRs mRNAs were widely expressed in various bovine tissues, but adiponectin mRNA expression was significantly higher in adipose tissue than in other tissues. In the mammary gland, although adiponectin mRNA expression was significantly decreased at lactation, AdipoR1 mRNA expression was significantly higher at peak lactation than at the dry‐off stage. In BMEC, lactogenic hormones and GH upregulated AdipoR2 mRNA expression but did not change that of AdipoR1. In conclusion, adiponectin and its receptor mRNA were expressed in various bovine tissues and the adiponectin mRNA level was decreased during lactation. These results suggest that adiponectin and its receptors ware changed in mammary glands by lactation and that AdipoRs mRNA expression was regulated by different pathways in BMEC.     

The forest vegetation and its differentiation under disturbance in a temperate mountain, China

Composition and structure of the main forest types, as well as the species changes under disturbance, were described as the representation of the forest vegetation in the north slope of Changbai mountain, northeast China. There were 3 forest zones from 600 m to 2,000 m asl, (1) broad-leaved-conifer mixed forest zone, 600–1,100 m asl, dominated byPinus koraiensis, Tilia amurensis, Acer mono, Ulmus japonica, (2) coniferous forest zone, 1,100–1,700 m asl, dominated byPicea jezoensis var.komarovii, P. koraiensis, Abies nephrolepis, and (3)Betula ermanii forest zone, 1,700–2,000 m asl, dominated byBetula ermanii, except for the east slope which was occupied byLarix olgensis. The evergreen component increased with elevation up to the upper limit of the coniferous forest zone (1,700 m), but disappeared in theB. ermanii zone. Species richness was significantly related to elevation. Towards higher elevation, the lapse rate of species-number was 1.6 species per 100 m, and it was only one species near the timberline. Species richness was reduced sharply under intensive disturbance, such as clear cutting, windfalletc. The species diversity was very low in the early stage of succession, and high in the later.     

Life history traits of leopard coralgrouper Plectropomus leopardus in the Okinawa Islands, southwestern Japan

The life history of the leopard coralgrouper Plectropomus leopardus was examined for the purpose of stock evaluation and to help maintain populations in the Okinawa Islands, southwestern Japan. Age was estimated from cut and burnt otoliths, and gonads were observed histologically to reveal the growth, spawning period and relationships between age and both sexual development and sexual maturation for P. leopardus in waters north of Okinawa Island. The three parameters in the von Bertalanffy growth equation, L _∞ , k, and t ₀, were estimated at 61.2 cm fork length, 0.289, and 0.41, respectively. The oldest individual obtained among the specimens was 18.8 years. The spawning period started in May and lasted until July. During this period, 50 % of females reached maturity at 43.3 cm fork length and at 5 years of age. Due to the sexual transition from female to male, the sex ratio decreased to 50 % at 59.8 cm fork length and at 10.3 years of age.     

A possible function of Nik-related kinase in the labyrinth layer of delayed delivery mouse placentas

In mice and humans, Nik-related protein kinase (Nrk) is an X-linked gene that encodes a serine/threonine kinase belonging to GCK group 4. Nrk knockout (Nrk KO) mice exhibit delayed delivery, possibly due to defective communication between the Nrk KO conceptus and its mother. However, the mechanism of delayed labor remains largely unknown. Here, we found that in pregnant mothers with the Nrk KO conceptus, the serum progesterone (P4) and placental lactogen (PL-2) concentrations in late pregnancy were higher than those in the wild type. Moreover, we demonstrated that Nrk is expressed in trophoblast giant cells (TGCs) and syncytiotrophoblast-2 (SynT-2) in the labyrinth layer of the mouse placenta. In the human placenta, NRK is also expressed in Syn-T in villi. Both human Syn-T and mouse TGCs of the labyrinth layer are present within fetal tissues that are in direct contact with the maternal blood. The labyrinth layer of the Nrk KO conceptus was gigantic, with enlarged cytoplasm and Golgi bodies in the TGCs. To investigate the function of Nrk in the labyrinth layer, a differentially expressed gene (DEG) analysis was performed. The DEG analysis revealed that labor-promoting factors, such as prostaglandins, were decreased, and pregnancy-maintaining factors, such as the prolactin family and P4 receptor, were increased. These findings suggest that the Nrk KO mice exhibit delayed delivery owing to high P4 concentrations caused by the hypersecretion of pregnancy-maintaining factors, such as PL-2, from the placenta.     

Review of diagnostic plaque reduction neutralization tests for flavivirus infection

Flavivirus infections (including Japanese encephalitis, West Nile encephalitis and dengue fever/severe dengue) present a worldwide public health problem. Recent climate change may affect the geographical distribution of the arthropod vectors for these viruses and so the risk of flavivirus epidemics may increase. Many methods have been developed for the serological diagnosis of flavivirus infections, such as haemagglutination inhibition assay, enzyme-linked immunosorbent assay, and immunofluorescence in staining. However, the specificity of these assays varies.The plaque reduction neutralizing test (PRNT) using live viruses is currently the ‘gold standard’ for the differential serodiagnosis of flaviviruses. The specificity of results obtained with PRNT is better than that for other protocols and many laboratories apply the PRNT protocol to the differential serodiagnosis of flaviviruses. Here, recent refinements to the PRNT protocols with genetically modified recombinant viruses or reporter-harbouring virus-like particles are reviewed. Further, the problems associated with the differential serodiagnosis of flaviviruses using PRNT are discussed.