Synthesis of Short-chain-length/Medium-chain-length Polyhydroxyalkanoate (PHA) Copolymers in Peroxisome of the Transgenic Arabidopsis Thaliana Harboring the PHA Synthase Gene from Pseudomonas sp. 61-3

In this paper, the photosynthetic production of short-chain-length/medium-chain-length polyhydroxyalkanoate (PHA) copolymers is reported. The wild-type and highly active doubly mutated PHA synthase 1 (S325T/Q481K, abbreviated ST/QK) genes from Pseudomonas sp. 61-3 were introduced into Arabidopsis thaliana. Peroxisome targeting signal 1 (PTS1) was used to target PHA synthases into the peroxisome to synthesize PHA from the intermediates of the β-oxidation pathway. The transgenic Arabidopsis produced PHA copolymers consisting of 40–57 mol% 3-hydroxybutyrate, 21–49 mol% 3-hydroxyvalerate, 8–18 mol% 3-hydroxyhexanoate, and 2–8 mol% 3-hydroxyoctanoate. The maximum PHA contents were 220μ g/g cell dry weight (cdw) in leaves, and 36μ g/g cdw in stems, respectively. The expression of the ST/QK mutated PHA synthase in leaves gene did not lead to significant difference in PHA content and monomer composition of PHAs, compared to the wild-type PHA synthase gene, suggesting that the supply of monomers may be a rate-determining step of PHA biosynthesis in the peroxisome. However, in stems, there were significant differences dependent on whether the wild-type or ST/QK mutated PHA synthase was expressed. These results suggest that tissue-specific monomer availability is important in determining the final mol% composition of PHA copolymers produced by the peroxisome in plants.     

Mg-Fe layered double hydroxide with chloride intercalated: synthesis,characterization and application for efficient nitrate removal

Environmental Science and Pollution Research - Mg-Fe layered double hydroxide intercalated with chloride (Mg-Fe-Cl LDH) was synthetized, characterized, and evaluated as adsorbent to remove nitrate...     

Risk assessment tools incorporating human error probabilities in the Japanese small-sized establishment

Although it has been estimated that as many as 80% of all occupational accidents have human errors as a cause, no risk assessment tools incorporating human-related elements have been developed for small companies. Human error probability (HEP) and human error analysis (HEA) have been used for large-scale, safety-critical industries for last three decades, but these tools are not suitable for smaller, more general industries that comprise the majority of accident settings.Here, we describe and verify a risk assessment tool that includes human-related elements for small companies. The tool expands on traditional risk assessment methods, such as matrix, risk graph and numerical scoring method, by adding human-related elements. The tool is easy-to-use in occupational environments, and includes assessments of human behavior and potentially outdated machinery at work place.     

Implication of nitric oxide in the heat-stress-induced cell death of the symbiotic alga <Emphasis Type="Italic">Symbiodinium microadriaticum</Emphasis>

One of the major consequences of global warming is a rise in sea surface temperature which may affect the survival of marine organisms including phytoplankton. Here, we provide experimental evidence for heat-induced cell death in a symbiotic microalga. Shifting Symbiodinium microadriaticum from 27 to 32°C resulted in an increase in mortality, an increase in caspase 3-like activity, and an increase in nitric oxide (NO) production. The caspase-like activity was strongly correlated with the production of NO in thermally challenged microalgae. For this experiment, the application of Ac-DEVD-CHO, a mammalian caspase 3-specific inhibitor, partly prevented (by 65%) the increase in caspase-like activity. To verify the relationship between NO and the caspase-like activity, S. microadriaticum were subsequently incubated with 1.0 mM of the following chemical NO donors: sodium nitroprusside (SNP), S-nitrosoglutathione (GSNO), S-nitroso-N-acetylpenicillamine (SNAP) and 3,3bis(Aminoethyl)-1-hydroxy-2-oxo-1-triazene (NOC-18). The supplementation of both SNP and NOC-18 caused a significant increase in caspase-like activity compared to the control treatment. Pre-treatment of the microalgae with the inhibitor Ac-DEVD-CHO before the supplementation of the different NO donors completely prevented the increase in caspase-like activity. These results suggest that NO could play a role in the induction of cell death in heat-stressed S. microadriaticum by mediating an increase in caspase-like activity.     

Possible involvement of the glycocalyx in the ichthyotoxicity of Chattonella marina (Raphidophyceae): immunological approach using antiserum against cell surface structures of the flagellate

Although the ichthyotoxic mechanism of Chattonella marina is still unknown, several lines of evidence suggest that the reactive oxygen species (ROS), such as superoxide anion (O₂^-), hydrogen peroxide (H₂O₂)_, and hydroxyl radical (·OH), produced by C. marina are involved in the mortality of fish exposed to this flagellate. Recently, we found that the cell-free supernatant prepared from C. marina, which is considered to contain the glycocalyx, showed NADPH-dependent O₂^- generation. In this study, we prepared antiserum against the crude glycocalyx of C. marina. Using indirect immunofluorescence, it was confirmed that the antiserum specifically reacted with C. marina cells. In addition to C. marina, the antiserum also reacted with other raphidophycean flagellates such as Heterosigma akashiwo, Olisthodiscus luteus, and Fibrocapsa japonica, whereas no reactivity was observed against six other flagellate species tested. These results suggest that raphidophycean flagellates have common epitopes recognized by the antiserum. Interestingly, immunohistochemical analysis of paraformaldehyde-fixed gill lamellae from yellowtail exposed to C. marina revealed that the antiserum stained the surface of gill lamellae, while no such staining pattern was observed in control gill lamellae. These results suggest that the glycocalyx may be discharged when C. marina cells are inhaled into the fishes' mouths and then come into contact with the gill surface. Based on the present results, together with our previous findings, we propose that continuous accumulation of the discharged glycocalyx on the gill surface occurs during C. marina exposure, which may be responsible for the ROS-mediated severe gill tissue damage leading to fish death.     

Spectroscopic analysis of NIR-dye sensitization in bulk heterojunction polymer solar cells

The photovoltaic conversion efficiency for near-infrared (NIR) sunlight is improved successfully by dye sensitization of bulk heterojunction polymer solar cells, in which the active layer was prepared by a ternary blend of poly(3-hexylthiophene), a fullerene derivative (1-(3-methoxycarbonyl)propyl-1-phenyl-[6,6]-methanofullerene), and an NIR dye, silicon phthalocyanine bis(trihexylsilyl oxide). The mechanism of the NIR-dye sensitization is studied by femtosecond transient absorption spectroscopy.     

Effects of biosurfactants on assays of PCB congeners in transgenic arabidopsis plants carrying a recombinant guinea pig AhR-mediated GUS reporter gene expression system

The transgenic Arabidopsis plants carrying a recombinant guinea pig (g) aryl hydrocarbon receptor (AhR)-mediated β-glucuronidase (GUS) reporter gene expression system were generated for assays of polychlorinated biphenyl (PCB) congeners. The selected transgenic Arabidopsis plant XgD2V11-6 exhibited a correlation between uptake of PCB126 and PCB126-induced GUS activity. Also, the plants showed induced GUS activity towards the supplemental indole 3-acetic acid (IAA). Thus, the GUS assay may reflect induction by both endogenous and exogenous AhR ligands. When biosurfactants, MEL-B, produced in the culture of yeast isolated from plants were used for assays of PCB congeners in the transgenic Arabidopsis plants, they showed marked PCB126 dose-dependent and toxic equivalency factor (TEF) dependent GUS activities. The effects of biosurfactants were clearer when the plants were cultivated on soils containing PCB congeners for 7 days as compared with on soils for 3 days as well as in the medium for 3 days. Threfore, it was estimated that biosurfactants form micellae with PCB congeners, which are easily uptaken by the plants in a mode of passive diffusion, transport into the aerial parts and then induce GUS activity.     

Spectroscopic Analysis of NIR-Dye Sensitization in Bulk Heterojunction Polymer Solar Cells

The photovoltaic conversion efficiency for near-infrared (NIR) sunlight is improved successfully by dye sensitization of bulk heterojunction polymer solar cells, in which the active layer was prepared by a ternary blend of poly(3-hexylthiophene), a fullerene derivative (1-(3-methoxycarbonyl)propyl-1-phenyl-[6,6]-methanofullerene), and an NIR dye, silicon phthalocyanine bis(trihexylsilyl oxide). The mechanism of the NIR-dye sensitization is studied by femtosecond transient absorption spectroscopy.