Dietary branched‐chain amino acid supplementation affects growth and hepatic insulin‐like growth factor gene expression in yellowtail,Seriola quinqueradiata

We investigated the effect of dietary supplementation with a branched‐chain amino acid (BCAA) mixture (valine, leucine and isoleucine) on juvenile yellowtail (Seriola quinqueradiata) growth and hepatic insulin‐like growth factor (IGF) gene expression. Total dietary BCAA content was 8.1, 8.5, 9.0 and 9.8 g   kg^?1 for the control, 0.4%, 1.0% and 2.0% BCAA experimental groups, respectively. Body weight was measured on day 77, after which fish were fasted for 3 days and refed; the livers were then removed 3 or 9 h later for analysis of hepatic gene expression. Body weight significantly increased in the BCAA‐supplemented groups. Hepatic expression of IGF1 and IGF2 significantly increased in the 2% BCAA group at 3 and 9 h after refeeding. In the 1% BCAA group, hepatic IGF1 expression tended to be higher at 9 h than in the control group after refeeding. Also, hepatic IGF2 significantly increased at only 9 h after refeeding in the 1% BCAA group. In conclusion, dietary supplementation with crystalline BCAAs increased growth and hepatic expression of IGF1 and IGF2. These results suggest that dietary crystalline BCAA supplementation would be a valuable addition to yellowtail aquaculture practices.     

Expression and localization of NO synthase isoenzymes (iNOS and eNOS) in development of the rabbit placenta

Nitric oxide synthase (NOS) is a key regulator of angiogenesis and embryogenesis in the mammalian reproductive process. Here, we attempted to clarify the expression and localization of inducible and endothelial NOS (iNOS and eNOS) in the developing rabbit placenta. Real-time RT-PCR analysis indicated that iNOS mRNA was significantly upregulated till the complete development of the placenta (d18), and then significantly decreased at the end of fetal growth stage (d28) during successful pregnancy. The eNOS mRNA was also enhanced in the pregnant uteri and gradually decreased near the term of pregnancy. Western blot analysis also showed elevation of the iNOS and eNOS protein levels during the course of successful pregnancy till the functional maturation of the placenta (d18). Immunohistochemical study revealed distinct localizations of iNOS along the radial arteries and eNOS at the spiral arteries and arterial sinuses in the developing placenta. This may reflect that iNOS and eNOS participate in pregnancy success through placentation-specific vascular formation and by supporting adequate blood circulation in the rabbit placenta.     

Gastric pleomorphic leiomyosarcoma in a Shetland sheepdog

A solitary mass of 6 x 6 x 5 cm was found in the smooth muscle layer of the pyloric part in the stomach of a Shetland sheepdog. The excised mass was solid, gray-white in color, and had central necrosis with hemorrhages. Microscopically, the neoplasm was composed of irregular bundles of spindle-shaped tumor cells and round tumor cells. A few multinucleated cells and gangliocyte-like cells were observed. The nuclei exhibited a high degree of atypia and many cells contained mitotic figures. Immunohistochemically, tumor cells were positive for vimentin and desmin, but negative for c-kit antibody. Ultrastructurally, tumor cells contained cytoplasmic myofilaments and pinocytotic vesicles. Based on these pathological findings, the tumor was diagnosed as gastric pleomorphic leiomyosarcoma.     

Extractable sulfate content in Japanese forest soils

The amounts of extractable sulfate in 12 forest soils were measured as a preliminary work to study sulfur dynamics in forest ecosystems. The sulfate content determined by the distillation method varied widely (10–880 × 10⁻⁶ gSg⁻¹) depending on the soil type and the depth. Japanese forest soils were broadly divided into two groups in sulfate level: one retains a large amount of sulfate and the other does not. In general, the surface soils to the depth of 10 cm contained small amounts of sulfate (< 100 × 10⁻⁶ gSg⁻¹), while subsoils contained more. The soil samples with low (< 4.6) or high (6 <) pH (H₂O) retained small amounts of sulfate. Black soils derived from volcanic ash retained great quantities of sulfate. The two levels of sulfate contents in Japanese forest soils suggests that the sulfur dynamics are different in these soils.     

Effects of bisphenol A (BPA) on placentation and survival of the neonates in mice

The effects of bisphenol A (BPA) on placentation have not been fully determined. The aim of this study was to clarify the structural changes of the placenta, abortion rate, and survival of neonates after BPA administration in mice. BPA (10 mg/kg/day) was administered to pregnant mice (BPA mice) subcutaneously from the first day of pregnancy (Day 0) to Day 7 (8 days total). The number of embryos and weights of whole uteri were measured on Days 10 and 12. Morphological changes in the placentae were examined by light microscopy on the corresponding days of pregnancy. The number of neonates was also counted. Survival rates were periodically calculated for neonates from the first day after parturition (P-Day 0) to P-Day 56. The number of embryos and weight of the uterus on Days 10 and 12 were significantly decreased by BPA injection. No notable differences were recognized between the left and right uteri. The proportion of the labyrinthine zone per whole placenta in the BPA mice became lower than that in the controls, and that of the metrial gland was higher in the BPA mice. The intervillous spaces of the placenta were narrower in the BPA mice. Degenerative changes were found in the trophoblastic giant cells and spongiotrophoblast layers of the BPA mice. The number of BPA mouse neonates was drastically decreased within 3 days after birth, and no mice survived after P-Day 56. The results suggest that BPA not only disrupts placental functions and leads to abortion through chronic stimulation of gene expression by binding to DNA but that it also affects the mortality of neonates through indirect exposure of embryos.     

The genetic mechanisms of warfarin resistance in Rattus rattus found in the wild in Japan

Warfarin is commonly used worldwide as a rodenticide. It inhibits blood coagulation by inhibiting vitamin K 2,3-epoxide reductase (VKOR) activity leading to hemorrhage. However, it has been reported that repeated or long-term treatment with warfarin results in resistance emerging in wild rodents. Such resistance may explain why it is difficult to control rodents in many regions in Japan. In this report, we studied mutations in the VKOR gene (including the VKOR complex subunit 1 (VKORC1)), while also analyzing VKOR and clotting factor activity in black rats (Rattus rattus) in order to understand better the mechanism of warfarin resistance in this species.We sequenced the VKORC1 gene from 275 rats living in the wild in Japan. We found several types of novel base substitutions, some of which conferred warfarin resistance.There was no difference in coagulation times between warfarin-sensitive and resistant rats measured under physiological conditions. However, after warfarin administration, no effect was noted in warfarin-resistant rats, although a prolonged coagulation time was noted in warfarin-sensitive rats.We also determined the kinetic differences in hepatic microsomal VKOR-dependent activity between warfarin-resistant and sensitive rats. Warfarin-resistant rats showed 2-3-fold lower V_max/K_m values than did sensitive rats. In addition, we report that resistant rats found in the Tokyo area had a VKOR activity which was poorly inhibited by warfarin.Finally, we conclude that reduced VKOR activity and warfarin resistance in the Japanese black rat might be due to mutations in the VKORC1 gene. However, further study is needed to clarify how such rats can maintain adequate vitamin K-dependent clotting factor levels, while simultaneously exhibiting low VKOR activity and warfarin resistance.     

Transcriptional profiling of inflammatory cytokine genes in African buffaloes (Syncerus caffer) infected with Theileria parva

Theileria parva (T. parva) causes East Coast fever (ECF), which is of huge economic importance to Eastern and Southern African countries. In a previous bovine model, inflammatory cytokines were closely associated with disease progression in animals experimentally infected with T. parva. The African Cape buffalo (Syncerus caffer), the natural reservoir for T. parva, is completely resistant to ECF despite a persistently high parasitaemia following infection with T. parva. Characterizing basic immunological interactions in the host is critical to understanding the mechanism underlying disease resistance in the African Cape buffalo. In this study, the expression level of several cytokines was analyzed in T. parva-infected buffaloes. There were no significant differences in the expression profiles of inflammatory cytokines between the infected and uninfected animals despite a remarkably high parasitaemia in the former. However, the expression level of IL-10 was significantly upregulated in the infected animals. These results indicate a correlation between diminished inflammatory cytokines response and disease resistance in the buffalo.     

Remnants of the early solar system water enriched in heavy oxygen isotopes

Oxygen isotopic composition of our solar system is believed to have resulted from mixing of two isotopically distinct nebular reservoirs, 16O-rich and (17,18)O-rich relative to Earth. The nature and composition of the (17,18)O-rich reservoir are poorly constrained. We report an in situ discovery of a chemically and isotopically unique material distributed ubiquitously in fine-grained matrix of a primitive carbonaceous chondrite Acfer 094. This material formed by oxidation of Fe,Ni-metal and sulfides by water either in the solar nebula or on a planetesimal. Oxygen isotopic composition of this material indicates that the water was highly enriched in 17O and 18O (delta(17,18)O(SMOW) = +180 per thousand per mil), providing the first evidence for an extremely (17,18)O-rich reservoir in the early solar system.     

Phosphorus application reduces N2O emissions from tropical leguminous plantation soil when phosphorus uptake is occurring

We examined the effects of phosphorus (P) fertilization on N₂O emissions from an Acacia mangium plantation in Indonesia. We focused on the roles of microbial and plant root activities using a trenching method to prepare root-excluded and root-including plots. In root-excluded plots, P application did not change the amount of N₂O emissions. By contrast, in root-including plots, P application significantly reduced N₂O emissions (from 71.1?±?20.2 to 19.3?±?5.1 mg N m^?2 106 days^?1). Lower total P, Bray-2 P, and Bio-P (microbial P determined by chloroform fumigation extraction method) contents in the soils of root-including plots as compared to root-excluded plots a few days after P application shows that acacia trees absorbed P fertilizer rapidly. This rapid P uptake probably relieved the P limitation of acacia and might have consistently increased root N uptake. This interpretation is supported by lower inorganic N content in P-applied soils (the average of three sampling times is 8.9 and 11.3 μg N g soil^?1 in P-applied soils and soils without P application, respectively), which in turn decreased N₂O emissions. Our study suggests that P fertilizer suppresses N₂O emissions from tropical leguminous forest plantations.