Synthesis of morphiceptin (Tyr-Pro-Phe-Pro-NH(2)) by dipeptidyl aminopeptidase IV derived from Aspergillus oryzae

Morphiceptin (Tyr-Pro-Phe-Pro-NH(2)), tetrapeptide, was synthesized using dipeptidyl aminopeptidase IV (DP IV, EC 3.4.14.5) derived from Aspergillus oryzae RIB 915 as a catalyst. Tyr-Pro-OEt was incubated with Phe-Pro-NH(2) in the presence of DP IV under various conditions of temperature, concentrations of ethylene glycol, pH, reaction time, and others. Morphiceptin was obtained at 40% yield under the optimal reaction conditions: substrate, 4 mM Tyr-Pro-OEt.HCl and 20 mM Phe-Pro-NH(2).HCl; enzyme, DP IV, 0.275 nkat; solvent, 60% ethylene glycol containing 20 mM phosphate buffer at pH 7.0; amine, 4.2 mM diisopropylamine at 4 degrees C for 24 h. Amino group protection was unnecessary for synthesis of morphiceptin by DP IV.     

Characterization of corrinoid compounds from a Japanese black tea (Batabata-cha) fermented by bacteria

A Japanese fermented black tea (Batabata-cha) contained a considerable amount of vitamin B(12) (456 +/- 39 ng per 100 g dry tea leaves and 2.0 +/- 0.3 ng per 100 mL of tea drink). A corrinoid compound was partially purified and characterized from the tea leaves. The patterns of the purified compound by the silica gel 60 thin-layer chromatography and C18 reversed phased high-performance liquid chromatography were identical to those of authentic vitamin B(12). When 20 week old vitamin B(12) deficient rats, which excreted substantial amounts (about 250 mg/day) of methylmalonic acid in urine as an index of vitamin B(12) deficiency, were fed the tea drink (50 mL/day, 1 ng of vitamin B(12)) for 6 weeks, urinary methylmalonic acid excretion (169 +/- 29 mg/day) of the tea drink-supplemented 26 week old rats decreased significantly relative to that (250 +/- 32 mg/day) of the deficient rats. The results indicate that the vitamin B(12) found in the fermented black tea is bioavailable in mammals.     

Humus composition of soils under forest, coffee and arable cultivation in hilly areas of south Sumatra, Indonesia

To understand the effect of land use changes on the composition of humus in tropical soils, samples from land under primary forest, secondary forest, coffee plantation, and arable crops were investigated at three sites in south Sumatra, Indonesia. Total carbon and total nitrogen contents were 1.7 to 4.3 times and 1.1 to 2.8 times greater in the topsoil under primary forest than under the other types of land use. Following change from primary forest to other uses, the proportion of humic acids in the organic matter of the topsoils decreased while that of the fulvic acid fraction increased. Within the range of land uses, differences in the yields of humic acids and fulvic acid fractions were, respectively, larger and smaller than those in total carbon content. The humic acids were classified into the low and middle classes in the degree of humification. Absorption due to the green fraction of humic acids, Pg, was detected in the UV‐visible spectra of almost all the humic acids. No relation was observed between the degree of humification of humic acids or the strength of Pg absorption in their spectra and land use change. The fulvic acid fractions were fractionated on insoluble polyvinylpyrrolidone (PVP) into the adsorbed fractions consisting of humic substances and the non‐adsorbed fractions consisting of non‐humic substances. A positive correlation between the amount of the fulvic acid fraction and the percentage of the PVP‐adsorbed fraction within it indicated that the variation in the amount of the fulvic acid fraction was attributable to acid‐soluble humic substances. The ionization difference spectra of solutions between pH 12 and pH 7 suggested that the chemical structures of the PVP‐adsorbed fulvic acids have been altered by land use change.     

Contribution of Parker and Product indexes to evaluate weathering condition of Yellow Brown Forest soils in Japan

Soil weathering is recognizable through study on physico-chemical characteristics of soil. In this study, Parker, ([(Na)_a / 0.35] + [(Mg)_a / 0.9] + [(K)_a / 0.25] + [(Ca)_a / 0.7]) × 100, and Product, [SiO₂ / (TiO₂ + Fe₂O₃ + SiO₂ + Al₂O₃)] × 100, weathering indexes, originally developed for study on rocks, were tested on Yellow Brown Forest soils from two different regions in Japan. The results showed that the contribution of the mentioned indexes can be used for study on soils weathering. In other word, in a two-dimensional coordination system where the axes are Parker and Product indexes, regarding to the chemical bases of the indexes, initial and lateral products of weathering can be tracked at the same time.

Additionally, the indexes relations with soil color were studied after exposing the samples at a high temperature. Remarkable relationships between a* / b* ratio–parameters of psychometric chromaticness of color in CIE color standard system–and these two weathering indexes demonstrated that a* / b* ratio can be applied reliably to estimate the quantities of Parker and Product indexes in Yellow Brown Forest soils.     

Comparison of Ni/Zn Accumulation Ability of Thlaspi japonicum from Three Different Areas in Hokkaido

Thlaspi japonicum is a Japanese Ni-hyperaccumulator plant growing in areas with serpentine soil containing a high level of Ni. In the present study, we investigated the ability of wild Thlaspi japonicum plants inhabiting non-ultramafic soil areas of Hokkaido to accumulate Ni/Zn. Wild T. japonicum plants growing on Rebun Island (Rebun-T) accumulated small amounts of Ni, while the plants cultivated on ultramafic rock soil accumulated large amounts of Ni, like T. japonicum growing on Mt. Yubari (Yubari-T), and this change was also observed for T. japonicum growing on Mt. Tengu (Tengu-T). All the T. japonicum plants transplanted on both ultramaflc soil or Andosol accumulated over 100 mg kg⁻¹ Zn, especially the Ni/Zn content of Tengu-T plants tended to be higher than that of the plants growing in the other two areas. In contrast, transplanting of Yubari-T plants from ultramafic soil with a high level of Ni to soil with a lower-Ni content decrease of the Ni concentration in the leaves. Wild Rebun-T plants showed a smaller biomass than the Yubari-T and Tengu-T plants, and some individuals of the Rebun-T population were smaller than those of the other populations, with tiny radical leaves. Zinc content of the small leaves was not significantly different from that in large leaves. These results suggested that T. japonicum plants are able to accumulate high Ni/Zn accumulation, regardless ecotype, habitat and leaf size.     

Characteristics of Raindrop Acidification by Co-Washout of SO2(G)-H2O2(G)-HNO3(G)

In order to investigate the acid rain formation under the coexistence of SO₂(g), H₂O₂(g), and HNO₃(g) in the air, a mathematical model has been built and some numerical simulations have been carried out with use of the model. The simulation reveals that SO₂(g) absorbed into a raindrop is released and then re-absorbed as the fall distance increases. The desorption and re-absorption processes of SO₂(g) are caused by: (1) the fact that the equilibrium concentration of H₂O₂(aq) and HNO₃(aq) in raindrops are much higher than that of SO₂(aq), and (2) the fact that the oxidation reaction rate of HSO₃ ^? with H₂O₂(aq) increases with H⁺ concentration in raindrops. The degree of acidification of the rainwater has been estimated by introducing a raindrop size distribution. The acidification is mainly caused by the adsorption of SO₂(g) in the usual case where the atmospheric concentration of SO₂(g) is much higher than that of HNO₃(g). With the increase in the atmospheric concentration of HNO₃(g), the concentration of H⁺ generated from SO₂(g) decreases and the contribution of HNO₃(g) to the generation of H⁺ becomes dominant.

     

Iron content of rat serum ferritin.

The serum ferritin concentration was significantly higher in female than in male rats, reflecting higher iron stores in females than in males. The mean iron/protein ratio of serum ferritin was 0.018+/-0.008 (SD) (microg of Fe/microg of protein) in female rats and 0.011+/-0.011 in male rats, being much lower than that of liver ferritin (0.233+/-0.014 in females and 0.227+/-0.020 in males). Iron loading of rats significantly increased serum ferritin concentration, but did not influence the iron content of serum ferritin. These results indicate that rat serum ferritin contains only a small amount of iron independent of body iron stores.     

Evaluation of First-Feeding Regimens for Larval Nassau Grouper Epinephelus straitus and Preliminary, Pilot-Scale Culture through Metamorphosis

Two 10-day hatchery experiments were conducted to evaluate s-type (Hawaiian strain) and ss-type (Thailand strain) rotifers Brachionus plicatilis and cryogenically preserved oyster Crassostrea gigas trochophores as first feeds for larval Nassau grouper Epinephelus striatus. Newly hatched grouper larvae were reared at densities of 11.2–20.8/L in 500-L tanks at 36–38 ppt salinity, 25–26 C, and under a 11-h light: 13-h dark photoperiod. Beginning on day 2 posthatching (d2ph), prey were maintained at a density of 20 individuals/mL, while phytoplankton (Nanochloropsis oculata) was maintained at 500 × 10³ cells/mL. In experiment 1, survival and growth were higher (P < 0.05) for fish fed small s-type rotifers (mean lorica length = 117 μm; fish survival = 7.96%) selected by sieving than for fish fed non-selected rotifers (mean lorica length = 161 μm; fish survival = 2.13%). These results demonstrated the advantage of small prey size and suggested that super-small (ss-type) rotifer strains would be beneficial. In experiment 2, three feeding regimens were compared: 1) ss-type rotifers (mean lorica length = 147 μm); 2) oyster trochophores (mean diameter = 50 μm) gradually replaced by ss-type rotifers from d5ph; and 3) a mixed-prey teatment of 50% oyster trochophores and 50% ss-type rotifers. Survival was higher (P < 0.05) for larvae fed mixed prey (15.6%) than for those fed rotifers (9.73%) or trochophores and rotifers in sequence (2.55%), which also showed the slowest growth. Oyster trochophores, although inadequate when used exclusively, enhanced survival when used in combination with rotifers, possibly by improving size selectivity and dietary quality. In a pilot-scale trial, larvae were cultured through metamorphosis in two 33.8-m³ outdoor tanks. Fertilized eggs were stocked at a density of 10 eggs/L and larvae were fed ss-type rotifers from d2ph-d20ph, newly hatched Artemia from d15ph-d18ph, 1-d-old Artemia nauplii from d18ph-d62ph. Survival on d62ph was 1.17%, with a total of 5,651 post-metamorphic juveniles produced.     

The genetic basis of host resistance to Bacillus anthracis in inbred mice

Inbred mice of the C3H/He (resistant) and C57BL/6 (susceptible) strains and their hybrids were used in experiments to investigate the genetic regulation of host resistance to B. anthracis. The susceptibility of F1 mice to a standard spore suspension was similar to that of the resistant parent and not intermediate between values for the parent strains. There were no differences in mortality rate between F1 and reciprocal cross mice. Analysis of the results indicated that resistance of mice to B. anthracis is regulated by a single dominant gene.     

Detection of bovine rhinotracheitis virus antibody by neutralizing test and ELISA in experimentally infected rabbits.

The antibody response of rabbit to infectious bovine rhinotracheitis (IBR) virus was examined. Two rabbits were inoculated with IBR virus strain Los Angeles into the trachea and intravenously, and intravenously two times, respectively. The patterns of antibody titers in the rabbits measured by neutralization test and ELISA were similar to those in the case of bovine. The antibody was detected after the inoculation, and much more antibody was detected after the second inoculation. The fact suggests that rabbits are a very useful laboratory animal for IBR virus infection studies.