Enzymatic synthesis of 4-amino-3,5-diethylphenyl sulfate, a rodent metabolite of alachlor.

Rat liver tissue homogenates were utilized for in vitro enzymatic conversion of 2,6-diethylaniline (DEA) to the important alachlor metabolite 4-amino-3,5-diethylphenyl sulfate (ADEPS), which was also generated as a radiolabeled standard for use in metabolism studies. ADEPS formation in rodents is associated with the production of other reactive metabolites implicated in alachlor rodent carcinogenesis, making dependable access to an ADEPS standard highly desirable. (14)C-DEA was oxidized by rat liver microsomes to (14)C-4-amino-3,5-diethylphenol, which was further converted to ADEPS via addition of the phosphosulfate transferase cofactor adenosine-3'-phosphate-5'-phosphosulfate. Microprobe NMR was used in conjunction with high-resolution mass spectrometry to fully characterize the resulting (14)C-ADEPS and confirm its structure. Because microgram quantities sufficed for full characterization, the enzymatic transformation provides a viable alternative to radiosynthesis of (14)C-ADEPS.     

The smell and odorous components of dried shiitake mushroom, Lentinula edodes VI: increase in odorous compounds of dried shiitake mushroom cultivated on bed logs

Odor is one of the most important characteristics affecting consumer preference for dried shiitake mushrooms [Lentinula edodes (Berk.) Pegler]. In our previous studies, we found that the odor content of commercial dried products was too weak for most people, and that the odorous compound content could be increased by adding amino acids to sawdust media. Currently, however, bed-log cultivation is used to produce fruiting bodies for dried products. The purpose of this study was to fi nd a method to increase the content of odorous compounds in dried products cultivated on bed logs. Pressure injection of amino acids from the side of the bed log was the most effi cient method, but it had some problems. Hence, a simpler and less troublesome method was developed, i.e., injecting amino acid solution from small bottles set in deep holes bored in the sides of the bed logs. In fruiting bodies cultivated on bed logs injected with amino acid solution by the improved method, the mean contents of lentinic acid, a precursor of the odorous compound lenthionine, approximately doubled compared to that in the untreated logs, although the infi ltration area of the solution injected by the improved method was smaller than that by the former method.     

Nickel-catalyzed carbonization of wood for coproduction of functional carbon and fluid fuels I: production of crystallized mesoporous carbon

Japanese larch wood loaded with nickel (1%–4%) alone or with nickel and calcium (0.25%–1.5%) was carbonized at 800°–900°C for 0–120min with a heating rate of 5°–20°C min⁻¹ in a helium flow of 5.8−46.4 ml STP cm⁻² min⁻¹ to examine the influence of these variables on the crystallization of carbon (the formation of T component) and the development of mesoporosity. From the obtained results, reaction conditions suitable for effective production of carbon with the dual functions of adequate electroconductivity and adsorption capacity in liquid phase were established, thereby explaining the factors that govern the process. It was also confirmed that mesopore having a diameter of about 4 nm was selectively produced at the cost of specific (BET) surface area in parallel with the formation of T component. This result provided good insight into how the simultaneous dual function could be realized.     

The effects of within-species and between-species variation in wood density on the photodegradation depth profiles of sugi (Cryptomeria japonica) and hinoki (Chamaecyparis obtusa)

Low density wood is more rapidly eroded than denser wood when exposed to the weather, possibly because it is more susceptible to photodegradation. Fourier transform infrared microscopy was used to examine: (1) the depth of photodegradation in earlywood and latewood of sugi (Japanese cedar) and earlywood of hinoki (Japanese cypress) exposed for up to 1500 h to artificial sunlight emitted by a xenon lamp (375 W/m² within the 300 to 700 nm spectral range); and (2) the relationship between the density of wood tissues and depth of photodegradation. The depth of photodegradation varied between species (sugi and hinoki) as well as within a growth ring (sugi earlywood and latewood), and there was an inversely proportional relationship between depth of photodegradation and wood density. These findings may explain why low density earlywood is more rapidly eroded than latewood during weathering, and more generally, why there is an inverse relationship between the density of wood species and their rate of erosion during artificial and natural weathering. Part of this work was presented at the 54th Annual Meeting of the Japan Wood Research Society, Sapporo, August 2004     

Microbial biomass and activity in urban soils contaminated with Zn and Pb

Summary The effects of heavy metals on microbial biomass and activity were investigated in 30 urban soils, contaminated mainly with Zn and Pb to different extents, in terms of the physicochemical and biological characteristics of the soils. Evaluated by simple and multiple regression analyses, the microbial biomass was not affected significantly by easily soluble Zn + Pb (extractable with 0.1 NHCI). The biomass was accounted for as a function of cation exchange capacity (CEC), total organic C and the numbers of fungal colonies present (R ² = 0.692). Carbon dioxide evolution from soils, which reflected microbial activity, was studied on soils incubated with microbial-promoting substrates (glucose and ammonium sulfate) or without. Carbon dioxide evolution was negatively related to Zn+Pb, and this inhibitory effect of the metals was greater in the soils incubated with substrates. Carbon dioxide evolution in soils with substrates was closely related to Zn+Pb, bacterial numbers and the numbers of fungal colonies (R ² = 0.718). Carbon dioxide evolution in soils without substrates was accounted for as a function of Zn + Pb, biomass and the C/N ratio (R ² = 0.511). Using these relationships, the effects of heavy metals on soil microorganisms are discussed in terms of metabolically activated and dormant populations.     

Sensitive enzyme immunoassay of cephalexin residues in milk, hen tissues, and eggs

A sensitive enzyme immunoassay for cephalexin (CEX) was developed using the rabbit antiserum to CEX, beta-D-galactosidase-labeled CEX, and a double-antibody separation method. The immunogen of CEX was prepared by coupling the amino group of CEX to thiol groups introduced into bovine serum albumin by the use of N-(m-maleimidobenzoyloxy)succinimide as a cross-linker. Highly titered antiserum to CEX was produced in rabbits immunized with the immunogen. Enzyme labeling of CEX with beta-D-galactosidase was done by using N-(gamma-maleimidobutyryloxy)succinimide as the cross-linker. The limit of detection was 30 ng CEX/mL sample solution. Application of the method to CEX drug residues detected 30 ng/mL in milk, 60 ng/g in egg yolk, and 400 ng/g in hen tissue.     

Effect of CO2 enrichment on fruit growth and quality in Japanese pear (Pyrus serotina Reheder cv. Kosui)

Six year-old Japanese pear (Pyrus seratina Reheder cv. Kosui) trees grafted on P. serotina cv. Nihonyamanashi were grown in containers filled with Granite Regosol under glasshouse conditions. At different stages of fruit growth, pear trees were exposed to an elevated CO₂ concentration (130 Pa CO₂ ) along with a control (35 Pa CO₂). For one group of plants, CO₂ enrichment was applied for 79 d from 52 d after full bloom (DAB) to fruit maturity (long-term CO₂ enrichment) and for another group the same treatment was applied for 35 d from 96 DAB to fruit maturity (short-term CO₂ enrichment). The effects of the elevated CO₂ concentration on vegetative growth, mineral contents, and fruit production and quality were examined. Long-term CO₂ enrichment enhanced vegetative growth, without any significant effect on the mineral contents in either flower bud or fruit except for a remarkable increase in the K content. Long-term CO₂ enrichment increased the fruit size and fresh weight, but had no significant effect on the fruit quality. On the other hand, the short-term CO₂ enrichment did not induce any significant change in the fruit size but increased the fruit sugar concentration. Along with the reduction of the sorbitol concentration in fruit, the fructose and sucrose concentrations increased and these changes occurred earlier at elevated CO₂ than at ambient CO₂ concentrations. From these results, we concluded that the effect of CO₂ enrichment on fruit growth varies depending upon the growth stages of fruit: during the initial and fruitlet stages when fruit expansion occurs, CO₂ enrichment increases the fruit size, whereas, during maturation when fruit expansion has slowed down and sugar accumulation in fruit is active, it increases the fruit sugar concentration.     

Cloning of the phloem-specific small heat-shock protein from leaves of rice plants

The N-terminal amino-acid sequence of a major rice phloem-sap protein, designated as RPP23, was determined. The complete amino-acid sequence of RPP23 was deduced from the corresponding rice EST- clone and contained an extra 46 amino acids at the N-terminus, that was apparently cleaved off to form mature RPP23 in sieve tubes. RPP23 shared a similarity to plant small heat-shock proteins (smHSPs), though the N-terminal region of RPP23 was distinct from that of known smHSPs. Immunocytological analyses using leaf sections showed that RPP23 was located only in the phloem regions of leaves, and was present in non-stressed plants. In mature leaves, stronger immunocytological signals were detected in sieve elements than in companion cells.     

Microbial mineralization of organic nitrogen into nitrate to allow the use of organic fertilizer in hydroponics

Hydroponics is an excellent technique for the cultivation of vegetable crops and other plants, but organic fertilizers cannot be used in conventional hydroponic systems, which generally use only inorganic fertilizers, because organic compounds in the hydroponic solutions generally have phytotoxic effects that lead to poor plant growth. Few microorganisms are present in hydroponic solutions to mineralize the organic compounds into inorganic nutrients. In this article a novel and practical hydroponic culture method that uses microorganisms to degrade organic fertilizer in the hydroponic solution has been developed. Soil microorganisms were cultured by regulating the amounts of organic fertilizer and inoculum, with moderate aeration. The microorganisms mineralized organic nitrogen via ammonification and nitrification into nitrate at an efficiency of 97.6%. The culture solution containing the microorganisms was usable as a hydroponic solution, and organic fertilizer could be directly added to it during vegetable cultivation. Vegetables grew well in the organic hydroponic system. Organic hydroponics based on this method is therefore a practical tool for the utilization of organic sources of fertilizer.