Increased expression of kisspeptin and GnRH forms in the brain of scombroid fish during final ovarian maturation and ovulation

BACKGROUND: Kisspeptins (Kiss) are prime players in the control of reproductive function through their regulation of gonadotropin-releasing hormone (GnRH) expression in the brain. The experimental scombroid fish, chub mackerel (Scomber japonicus) expresses two kiss (kiss1 and kiss2) and three gnrh (gnrh1, gnrh2, and gnrh3) forms in the brain. In the present study, we analyzed expression changes of kiss and gnrh mRNAs in the brain and corresponding GnRH peptides in the brain and pituitary during final ovarian maturation (FOM) and ovulation. METHODS: Female fish possessing late vitellogenic oocytes were injected with GnRH analogue to induce FOM and ovulation. Fish were observed for daily spawning activities and sampled one week post-injection at germinal vesicle migration (GVM), oocyte hydration, ovulation, and post-ovulatory time periods. Changes in relative mRNA levels of kiss and gnrh forms in the brain were determined using quantitative real-time PCR. Changes in GnRH peptides in the brain and pituitary were analyzed using time-resolved fluoroimmunoassay. RESULTS: Both kiss1 and kiss2 mRNA levels in the brain were low at late vitellogenic stage and increased significantly during the GVM period. However, kiss1 mRNA levels decreased during oocyte hydration before increasing again at ovulatory and post-ovulatory periods. In contrast, kiss2 mRNA levels decreased at ovulatory and post-ovulatory periods. Levels of gnrh1 mRNA in the brain increased only during post-ovulatory period. However, levels of gnrh2 and gnrh3 mRNAs were elevated during GVM and then, decreased during oocyte hydration before increasing again at ovulatory period. During post-ovulatory period, both gnrh2 and gnrh3 mRNA levels declined. Peptide levels of all three GnRH forms in the brain were elevated during GVM and oocyte hydration; their levels were significantly lower during late vitellogenic, ovulatory, and post-ovulatory periods. In contrast, pituitary GnRH peptide levels did not show any significant fluctuations, with the GnRH1 peptide levels being many-fold higher than the GnRH2 and GnRH3 forms. CONCLUSION: The results indicate increased expression of multiple Kiss and GnRH forms in the brain and suggest their possible involvement in the regulation of FOM and ovulation in captive female chub mackerel

Activation of invariant natural killer T cells stimulated with microbial α-mannosyl glycolipids

Some synthetic and bacterial glycolipids presented by CD1d specifically activate invariant NKT (iNKT) cells bearing an invariant Vα14-Jα18 (mouse) or Vα24-Jα18 (human) TCR. The antigenic glycolipids identified to date consist of two hydrophobic chains and an α-glycoside in which the 2′-OH group is in the cis orientation toward the anomeric group, namely, either an α-galactoside or an α-glucoside. Several microbial α-mannosyl glycolipids, in which the 2′-OH group is in the trans orientation, were herein examined to establish whether they have potential to activate iNKT cells. We found that α-mannnosyl1-3 (6′-O-acyl α-mannosyl)-1-1 monoacylglycerol and cholesteryl 6′-O-acyl α-mannoside, found in Saccharopolyspora and Candida albicans, respectively, induced the activation of iNKT cells, dependent on CD1d. In contrast, α-mannosyldiacylglycerol found in Streptococcus suis or α-mannosylceramide demonstrated markedly less antigenicity for iNKT cells. The potentially antigenic α-mannosyl glycolipids contributed to the protection of mice against infection with S. pneumoniae in which iNKT cells have previously been found to participate. Furthermore, these glycolipids induced the production of proinflammatory cytokines by macrophages, thereby suggesting their recognition by specific pattern recognition receptors (PRRs). Collectively, these results suggest that these microbial α-mannosyl glycolipids are capable of being recognized by both the invariant TCR and PRRs and inducing immune responses

Analysis of Expressed Sequence Tags from the Fungus Aspergillus oryzae Cultured Under Different Conditions

We performed random sequencing of cDNAs from nine biologically or industrially important cultures of the industrially valuable fungus Aspergillus oryzae to obtain expressed sequence tags (ESTs). Consequently, 21 446 raw ESTs were accumulated and subsequently assembled to 7589 non-redundant consensus sequences (contigs). Among all contigs, 5491 (72.4%) were derived from only a particular culture. These included 4735 (62.4%) singletons, i.e. lone ESTs overlapping with no others. These data showed that consideration of culture grown under various conditions as cDNA sources enabled efficient collection of ESTs. BLAST searches against the public databases showed that 2953 (38.9%) of the EST contigs showed significant similarities to deposited sequences with known functions, 793 (10.5%) were similar to hypothetical proteins, and the remaining 3843 (50.6%) showed no significant similarity to sequences in the databases. Culture-specific contigs were extracted on the basis of the EST frequency normalized by the total number for each culture condition. In addition, contig sequences were compared with sequence sets in eukaryotic orthologous groups (KOGs), and classified into the KOG functional categories

Development of Ultra-Micro Combustor Using Cylindrical Flames

The purpose of this study is to develop an ultra-micro combustor that uses two types of coaxial cylindrical flames, rich premixed flame and diffusion flame. The combustor consists of inner and outer porous tubes, and rich propane-air mixture and air issued, respectively, through the inner tube outwardly and through the outer tube inwardly, forming a cylindrical stagnation plane sandwiched by the inner rich premixed flame and the outer diffusion flame. Petal type flame was also observed in the downstream of the cylindrical flames. Keeping the equivalence ratio φi and flow rate qi of the rich mixture constant, air flow rate qa was varied. The O2 and CO concentrations and temperature of the burnt gas were measured, and heat loss rate ηhl and combustion intensity L were evaluated. The obtained results are described as follows. (1)The relation curve of ηhl with the overall equivalence ratio φall, which is evaluated from the total flow rate of the fuel and the air, has a minimum value. (2)The relation curve of the minimum value of ηhl with L has a minimum value. (3) The CO concentration of the burnt gas increases as qa is increased because of local extinction of the petal type flame. (4)When qa is increased further, petal type flame is also extinguished. After that, the O2 concentration increases and the CO concentration decreases

INTERACTIVE DYNAMICS OF FRUIT AND STEM GROWTH IN TOMATO PLANTS AS AFFECTED BY ROOT WATER CONDITION I. EXPANSION AND CONTRACTION OF FRUIT AND STEM

For on-line and intact measurement of fruit and stem growth, a laser displacement sensor (LDS) system was newly developed, and two sets of the LDS system were applied for dynamic analysis of fruit and stem growth in tomato plants (Lycopersicon esculentum Mill.) as affected by change in root water condition. The plants potted with soil were dewatered gradually for three days and then rewatered. Fruit and stem growth appeared in the dissymmetrical relationship: During dewatering, fruit growth was kept higher even under larger decrease in xylem water potential and significant stem contraction, but just after rewatering drastic fruit shrinkage was induced with rapid swelling of the stem. This demonstrates the interaction between fruit and stem growth through dynamics of water relations along the translocation path in stem. Thus, the newly developed LDS system can be reliable for dynamic analysis of fruit and stem growth