Contribution of dissolved organic nitrogen deposition to nitrogen saturation in a forested mountainous watershed in Tsukui, Central Japan

Nitrogen (N) budget was estimated with dissolved inorganic N (DIN) and dissolved organic N (DON) in a forested mountainous watershed in Tsukui, Kanagawa Prefecture, about 50 km west of Tokyo in Central Japan. The forest vegetation in the watershed was dominant by Konara oak (Quercus serrata) and Korean hornbeam (Carpinus tschonoskii), and Japanese cedar (Cryptomeria japonica). Nitrate (NO₃ ^?) concentration in the watershed streamwater was averagely high (98.0 ±± 19 (±± SD, n = 36) μmol L^?1) during 2001–2003. There was no seasonal and annual changes in the stream NO^? ₃ concentration even though the highest N uptake rate presumably occurred during the spring of plant growing season, a fact indicating that N availability was in excess of biotic demands. The DON deposition rates (DON input rates) in open area and forest area were estimated as one of the main N sources, accounting for about 32% of total dissolved N (TDN). It was estimated that a part of the DON input rate contributed to the excessive stream NO^? ₃ output rate under the condition of the rapid mineralization and nitrification rates, which annual DON deposition rates were positively correlated with the stream NO₃ ^? output rates. The DON retention rate in the DON budget had a potential capacity, which contributed to the excessive stream NO^? ₃ output rate without other N contributions (e.g. forest floor N or soil N).     

Distribution and Fate of Organotin Compounds in Japanese Coastal Waters

In order to elucidate the details of both the distribution and fate of organotin compounds (OTs) in the costal ecosystem, the concentrations of butyltin compounds (BTs) and phenyltin compounds (PTs) were determined in seawater, sediment and blue mussels Mytilus galloprovincialis collected in Maizuru Bay, Japan. The concentrations of tributyltin (TBT) in seawater, sediments and mussels ranged from 3.9 to 27 ng l^?1, from 1.2 to 19 ng g^?1 dry wt and from 0.77 to 11 ng g^?1 wet wt, respectively. Although the levels of TBT in seawater, sediments and mussels from Maizuru Bay were lower than those reported previously at other sites in Japan, the levels can still be toxic to susceptible organisms. Trace amounts of PTs were also found in seawater, sediment and mussel samples, indicating that there is a slight input of triphenyltin (TPT) into the seawater in the bay at present. The highest TBT concentration in seawater among all sites measured in the bay was found near a glass factory, and the lowest concentrations were observed at the center of the bay. The highest concentration of TBT in sediment was detected near a shipyard. In mussels, a high concentration of TBT was detected near a cement plant and timberyard. These results indicate that the major sources of contamination of OTs in the bay are considered to be from those facilities and ships. The proportion of TBT in seawater and mussels showed that, of total BTs, it was the predominant compound at most sites. These results suggested that there is a continuous input of TBT into the seawater and thereafter an accumulation of TBT in mussels due to their low metabolic capacity to degrade TBT. In sediment, the proportion of monobutyltin (MBT) was the highest of BTs at most sites. This could reflect a previous contamination by TBT used before the regulation of its usage in the bay.     

Cecal bacterial communities in wild Japanese rock ptarmigans and captive Svalbard rock ptarmigans

Preservation of indigenous gastrointestinal microbiota is deemed to be critical for successful captive breeding of endangered wild animals, yet its biology is poorly understood. Here, we investigated cecal bacterial communities in wild Japanese rock ptarmigans (Lagopus muta japonica) and compared them with those in Svalbard rock ptarmigans (L. m. hyperborea) in captivity. Ultra-deep sequencing of 16S rRNA gene indicated that the community structure of cecal microbiota in wild rock ptarmigans was remarkably different from that in captive Svalbard rock ptarmigans. Fundamental differences between bacterial communities in the two groups of birds were detected at the phylum level. Firmicutes, Actinobacteria, Bacteroidetes and Synergistetes were the major phyla detected in wild Japanese rock ptarmigans, whereas Firmicutes alone occupied more than 80% of abundance in captive Svalbard rock ptarmigans. Furthermore, unclassified genera of Coriobacteriaceae, Synergistaceae, Bacteroidaceae, Actinomycetaceae, Veillonellaceae and Clostridiales were the major taxa detected in wild individuals, whereas in zoo-reared birds, major genera were Ruminococcus, Blautia, Faecalibacterium and Akkermansia. Zoo-reared birds seemed to lack almost all rock ptarmigan-specific bacteria in their intestine, which may explain the relatively high rate of pathogenic infections affecting them. We show evidence that preservation and reconstitution of indigenous cecal microflora are critical for successful ex situ conservation and future re-introduction plan for the Japanese rock ptarmigan.     

Polyphenol composition of a functional fermented tea obtained by tea-rolling processing of green tea and loquat leaves

Phenolic constituents of a new functional fermented tea produced by tea-rolling processing of a mixture (9:1) of tea leaves and loquat leaves were examined in detail. The similarity of the phenolic composition to that of black tea was indicated by high-performance liquid chromatography comparison with other tea products. Twenty-five compounds, including three new catechin oxidation products, were isolated, and the structures of the new compounds were determined to be (2R)-2-hydroxy-3-(2,4,6-trihydroxyphenyl)-1-(3,4,5-trihydroxyphenyl)-1-propanone 2-O-gallate, dehydrotheasinensin H, and acetonyl theacitrin A by spectroscopic methods. In addition, theacitrinin A and theasinensin H were obtained for the first time from commercial tea products. Isolation of these new and known compounds confirms that reactions previously demonstrated by in vitro model experiments actually occur when fresh tea leaves are mechanically distorted and bruised during the production process.     

Nematocidal quassinoids and bicyclophosphorothionates: a possible common mode of action on the GABA receptor

The present study was undertaken to identify noncompetitive γ-aminobutyric acid receptor (GABAR) antagonists that are effective in nematodes, as well as to examine the hypothesis that the noncompetitive antagonism of the GABAR underlies the nematocidal activity of quassinoids against free-living nematodes of the Diplogastridae family. First, 14 known GABAR antagonists were screened for the effectiveness of their nematocidal activity in Diplogastridae. As a result, 3-isopropyl-1-phospha-2,6,7-trioxabicyclo[2.2.2]octane 1-sulfides (3-isopropyl-BPTs) were found to have high nematocidal activities, and 4-cyclohexyl-3-isopropyl-BPT (23) and 3-isopropyl-4-(2-propenyl)-BPT (27) were the two most potent analogues; these compounds are equipotent to samaderine B and more potent than the anthelmintic abamectin. 23-resistant nematodes, selected by challenge with 23, showed cross-resistance to samaderine B. 23 (10 μM) reduced [³H]23 binding to nematode membranes by 30.4%. Samaderine B (10 μM) resulted in a similar level of the inhibition, but had neither additive nor synergistic effects on the 23 inhibition of [³H]23 binding. These findings suggest that samaderine B shares a common binding site with the GABAR antagonist 23 in Diplogastridae. The results of comparative molecular field analysis, a method of three-dimensional quantitative structure-activity relationship analysis, supported this conclusion.     

Successful mouse cloning of an outbred strain by trichostatin A treatment after somatic nuclear transfer

Although the somatic cloning technique has been used for numerous applications and basic research of reprogramming in various species, extremely low success rates have plagued this technique for a decade. Further in mice, the "clonable" strains have been limited to mainly hybrid F1 strains such as B6D2F1. Recently, we established a new efficient cloning technique using trichostatin A (TSA) which leads to a 2-5 fold increase in success rates for mouse cloning of B6D2F1 cumulus cells. To further test the validity of this TSA cloning technique, we tried to clone the adult ICR mouse, an outbred strain, which has never been directly cloned before. Only when TSA was used did we obtain both male and female cloned mice from cumulus and fibroblast cells of adult ICR mice with 4-5% success rates, which is comparable to 5-7% of B6D2F1. Thus, the TSA treatment is the first cloning technique to allow us to successfully clone outbred mice, demonstrating that this technique not only improves the success rates of cloning from hybrid strains, but also enables mouse cloning from normally "unclonable" strains.     

Mouse in vivo-derived late 2-cell embryos have higher developmental competence after high osmolality vitrification and −80°C preservation than IVF or ICSI embryos

Mammalian embryos are most commonly cryopreserved in liquid nitrogen; however, liquid nitrogen is not available in special environments, such as the International Space Station (ISS), and vitrified embryos must be stored at −80°C. Recently, the high osmolarity vitrification (HOV) method was developed to cryopreserve mouse 2-cell stage embryos at −80°C; however, the appropriate embryo is currently unknown. In this study, we compared the vitrification resistance of in vivo-derived, in vitro fertilization (IVF)-derived, and intracytoplasmic sperm injection (ICSI)-derived mouse 2-cell embryos against cryopreservation at −80°C. The ICSI embryos had lower survival rates after warming and significantly lower developmental rates than the in vivo and IVF embryos. Further, IVF embryos had a lower survival rate after warming, but a similar rate to the in vivo embryos to full-term development. This result was confirmed by simultaneous vitrification of in vivo and IVF embryos in the same cryotube using identifiable green fluorescent protein-expressing embryos. We also evaluated the collection timing of the in vivo embryos from the oviduct and found that late 2-cell embryos had higher survival and developmental rates to full-term than early 2-cell embryos. Some early 2-cell embryos remained in the S-phase, whereas most late 2-cell embryos were in the G2-phase, which may have affected the tolerance to embryo vitrification. In conclusion, when embryos must be cryopreserved under restricted conditions, such as the ISS, in vivo fertilized embryos collected at the late 2-cell stage without long culture should be employed.