Biochemical and Genetic Characterization of an Extracellular Poly(3-Hydroxybutyrate) Depolymerase from Acidovorax Sp. Strain TP4

To determine the properties of enzymes from bacteria that degrade polypropiolactone (PPL), we isolated 13 PPL-degrading bacteria from pond water, river water, and soil. Nine of these strains were identified as Acidovorax sp., three as Variovorax paradoxus, and one as Sphingomonas paucimobilis. All the isolates also degraded poly(3-hydroxybutyrate) (PHB). A PPL-degrading enzyme was purified to electrophoretical homogeneity from one of these bacteria, designated Acidovorax sp. TP4. The purified enzyme also degraded PHB. The molecular weight of the enzyme was estimated as about 50,000. The enzyme activity was inhibited by diisopropylfluorophosphate, dithiothreitol, and Triton X-100. The structural gene of the depolymerase was cloned in Escherichia coli. The nucleotide sequence of the cloned DNA fragment contained an open reading frame (1476 bp) specifying a protein with a deduced molecular weight of 50,961 (491 amino acids). The deduced overall sequence was very similar to that of a PHB depolymerase of Comamonas acidovorans YM1609. From these results it was concluded that the isolated PPL-degrading enzyme belongs to the class of PHB depolymerases. A conserved amino acid sequence, Gly-X₁-Ser-X₂-Gly (lipase box), was found at the N-terminal side of the amino acid sequence. Site-directed mutagenesis of the TP4 enzyme confirmed that ²⁰Ser in the lipase box was essential for the enzyme activity. This is the first report of the isolation a PHB depolymerase from Acidovorax.     

PCDDs/DFs emissions from crematories in Japan

Concentrations of PCDDs and PCDFs in emission gases from 10 crematories were measured. The relationship between PCDDs/DFs and several factors such as structure, equipment and operational state of the crematory is discussed. Furthermore, emission of PCDDs/DFs from all crematories in Japan is estimated. The following results are obtained: (1) total concentration of PCDDs/DFs was 2.2-290 ng/N m3, whose TEQ concentration was 0.0099-6.5 ng TEQ/N m3; (2) total concentration of PCDFs was higher than that of PCDDs; (3) T4CDFs was the highest in the homologue pattern and 2,3,7,8-T4CDF was the highest in the isomer pattern; (4) emission of PCDDs/DFs was the largest in the first 20 min of cremation; (5) concentration of PCDDs/DFs was related to the existence of a secondary combustion chamber and a dust collector, and the ratio of the numbers of main and secondary combustion chambers; (6) total emission of PCDDs/DFs from crematories in Japan was estimated to be 8.9 g TEQ/yr.     

Validation of diffusive mini-samplers for aldehyde and VOC and its feasibility for measuring the exposure levels of elementary school children

Exposure to various chemicals can cause adverse effects to health, such as asthma and allergies, especially in children. Data on personal exposure levels in children are scarce, thus small lightweight diffusive mini-samplers for aldehydes and volatile organic compounds (VOCs) were designed to measure the exposure level of children to these chemicals. The aim of the study was to validate and examine the applicability of these mini-samplers for measuring daily chemical exposure. The diffusive mini-samplers are 20 mm in length, 11 mm in diameter, and 1.67 g in weight. The devices are cylindrically shaped with polytetrafluoroethylene membrane filters placed at each end. To measure aldehydes and acetone, 20 mg of 2,4-dinitrophenylhydrazine was used as an absorbent. To measure VOCs, a carbon molecular sieve was used. The sampling rate for each chemical was determined by parallel sampling with active samplers in a closed exposure bag. The blank levels of the chemicals and the storage stability of the device were tested. The mini-samplers were compared to commercially available diffusive samplers. To examine the applicability of the samplers, 65 elementary school children carried them for 24 h. The sampling rates for formaldehyde, acetaldehyde, and acetone were 20.9, 22.9, and 19.7 mL min(-1), respectively. The limits of quantification (LOQ) for the 24-hour sampling by high-performance liquid chromatography/ultraviolet (HPLC/UV) analysis were 8.3, 7.6, and 8.8 μg m(-3) for formaldehyde, acetaldehyde, and acetone, respectively. The sampling rates for the 11 VOCs were determined and ranged from 3.3 mL min(-1) for styrene and 2-ethyl-1-hexanol to 11.7 mL min(-1) for benzene. The LOQ for the 24-hour sampling by gas chromatography-mass spectrometry (GC-MS) analysis ranged from 5.9-105.2 μg m(-3), 1.1-24.7 parts per billion. The storage stability after 5 days ranged from 94.8 to 118.2%. Formaldehyde, acetone, benzene, and toluene were detected above the LOQ in more than 90% of the children, and the median concentrations were 21.7, 20.9, 10.1, and 21.5 μg m(-3), respectively. This study shows that the diffusive samplers developed were suitable for children to carry and were capable of measuring the children's daily chemical exposure.     

Inundation Analysis in the Coastal Area Considering Climate Change Due to Global Warming

This paper describes the possible inundation scenarios under sea level rise conditions due to global climate change with particular reference to Nagoya, Japan. The study was carried out by using a two-dimensional sea model integrated with one-dimensional river flow model and two-dimensional overland flow model. For the connections of models, the upstream discharge or downstream water level in each grid is considered as the boundary conditions. The governing equations used for the analysis have been solved by finite volume method. The analysis results implicate that some parts of densely populated coastal area of Nagoya city will be vulnerable to inundation if the sea level rise due to global warming by 1 m. Moreover, the performances of existing sewer system and inundation scenario under various conditions have been analyzed.     

Genotoxic potentials and related mechanisms of bisphenol A and other bisphenol compounds: A comparison study employing chicken DT40 cells

Bisphenol A (BPA) has been found in plastic food containers, paper currencies and toys. BPA has been reported for various adverse health concerns including reproduction, development and carcinogenesis. These potential health implications have led to increasing use of alternative bisphenols such as bisphenol F and bisphenol S among many. However, little is known about the toxicity of alternative bisphenols and most of the toxicological information is limited to endocrine disrupting potentials. In this study, we evaluated cytotoxicity and the genotoxic potentials of several bisphenol compounds, and identified the mechanism of genotoxicity using a panel of mutant chicken DT40 cell lines deficient in DNA repair pathways. Several bisphenols including bisphenol AP, bisphenol M, or bisphenol P exerted genotoxic potentials that are greater than that of BPA. Generally RAD54^−/− mutant cells were the most sensitive to all bisphenols except for bisphenol F, suggesting the induction of DNA double-strand breaks that could be rescued by homologous recombination. Genotoxic potential of bisphenols was confirmed by chromosomal aberration assay and γ-H2AX foci forming assay between wild-type and RAD54^−/− mutant. Among the tested bisphenols, BPP at 12.5 μM showed the greatest genotoxic potency, inducing chromosomal aberration and γ-H2AX foci in RAD54^−/− mutant by 2.6 and 4.8 folds greater than those in wild-type, respectively. Our results clearly show several alternative bisphenols can cause genotoxicity that could be rescued by homologous recombination pathway, and some bisphenols induced even greater genotoxic potentials than that of BPA.     

Production of 2-Pyrrolidone from Biobased Glutamate by Using Escherichia coli

We have developed a simple and highly efficient process for the production of 2-pyrrolidone (2-PRN) from biobased l-glutamic acid (Glu). First, we produced γ-aminobutyric acid (GABA) from Glu obtained by fermentation of biomass using Escherichia coli, which is known to possess GABA producing activity. The reaction solution contained only the substrate Glu, bacterial cells, and water, and did not require buffers or coenzymes, pyridoxal-5′-phosphate (PLP). Every 24 h, cells were removed by centrifugation, and GABA containing supernatant was obtained. This reaction can be repeated 14 times by adding water and Glu, without any decrease in activity. Finally, 303.7 g of GABA was produced from 560 g (40 g × 14 times) of Glu with a yield of 77.4 %. The concentration of this solution was almost 40 %. The GABA was then converted to biobased 2-PRN by heating and distillation under reduced pressure without pretreatment. The yield obtained with this chemical process was 95.8 %. These results showed that biobased 2-PRN could be produced from biomass-derived Glu. Biobased 2-PRN has great potential as a raw material to change other petroleum-based materials to biobased materials.     

Uptake and distribution of iodine in rice plants

Rice (Oryza sativa L.) plants were cultivated in an experimental field and separated at harvest into different components, including polished rice, rice bran, hull, straw, and root. The contents of iodine in these components and the soil were determined by inductively coupled plasma-mass spectrometry and radiochemical neutron activation analysis, respectively. Iodine content varied by more than three orders of magnitude among the plant components. Mean concentration of iodine in the entire plants was 20 mg kg(-1) dry weight, and the concentration of iodine in the surface soil (0-20 cm depth) was 48 mg kg(-1). The highest concentration of iodine (53 mg kg(-1) dry weight) was measured in root and the lowest concentration (0.034 mg kg(-1) dry weight) in polished rice. While the edible component (polished rice) accounted for 32% of the total dry weight, it contained only 0.055% of iodine found in the entire rice plants. Atmospheric gaseous iodine (5.9 ng m(-3)) was estimated to contribute <0.2% of the total iodine content in the biomass of rice plants; therefore nearly all of the iodine in the rice plants was a result of the uptake of iodine from the soil. The content of iodine in the aboveground part of rice plants was 16 mg kg(-1) dry weight and the percentage of iodine transferred per cropping from the soil into the aboveground biomass corresponded to 0.27% (20 mg m(-2)) of the upper soil layer content.     

Ecological effects of various toxic agents on the aquatic microcosm in comparison with acute ionizing radiation

The purpose of this study was an evaluation of the effect levels of various toxic agents compared with acute doses of ionizing radiation for the experimental model ecosystem, i.e., microcosm mimicking aquatic microbial communities. For this purpose, the authors used the microcosm consisting of populations of the flagellate alga Euglena gracilis as a producer, the ciliate protozoan Tetrahymena thermophila as a consumer and the bacterium Escherichia coli as a decomposer. Effects of aluminum and copper on the microcosm were investigated in this study, while effects of gamma-rays, ultraviolet radiation, acidification, manganese, nickel and gadolinium were reported in previous studies. The microcosm could detect not only the direct effects of these agents but also the community-level effects due to the interspecies interactions or the interactions between organisms and toxic agents. The authors evaluated doses or concentrations of each toxic agent which had the following effects on the microcosm: (1) no effects; (2) recognizable effects, i.e., decrease or increase in the cell densities of at least one species; (3) severe effects, i.e., extinction of one or two species; and (4) destructive effects, i.e., extinction of all species. The resulting effects data will contribute to an ecological risk assessment of the toxic agents compared with acute doses of ionizing radiation.     

Uptake and distribution of 90Sr and stable Sr in rice plants

The stable Sr content in the aboveground parts of rice plants at various growth stages, and the distributions of 90Sr and stable Sr in rice plant components, such as polished rice, rice bran, hull, straw and root, at harvest time, were determined. The total Sr content in the aboveground rice plants was dependent on the growth stage and followed the sigmoidal shape of the growth curve. The concentration of 90Sr among the different components of rice plants varied within two orders of magnitude, whereas the 90Sr/Sr concentration ratio had a constant value. Therefore, the translocation rate of 90Sr in rice plants had similar values to that of stable Sr. However, the 90Sr/Sr concentration ratio for the rice plants was different for each study site. Only 0.6% of the total Sr was found in polished rice, while more than 99% was found in the non-edible components, of which 87% was present in the straw. These findings suggest that 90Sr in the non-edible parts could have been transferred to humans through the soil-plant system and/or feed-livestock pathway. The soil-to-plant transfer factor of 90Sr in polished rice was 0.0021 +/- 0.00007, which was two orders of magnitude lower than that in the straw. The percentage of 90Sr removed from the upper soil layer to the aboveground biomass of rice plants at harvest time was calculated as 0.094%. It is possible that approximately 0.1% of the total 90Sr content in the surface soil layer is removed from the soil-plant system by human activities every year.