Genetic engineering of cyanobacteria to enhance biohydrogen production from sunlight and water

To mitigate global warming caused by burning fossil fuels, a renewable energy source available in large quantity is urgently required. We are proposing large-scale photobiological H(2) production by mariculture-raised cyanobacteria where the microbes capture part of the huge amount of solar energy received on earth's surface and use water as the source of electrons to reduce protons. The H(2) production system is based on photosynthetic and nitrogenase activities of cyanobacteria, using uptake hydrogenase mutants that can accumulate H(2) for extended periods even in the presence of evolved O(2). This review summarizes our efforts to improve the rate of photobiological H(2) production through genetic engineering. The challenges yet to be overcome to further increase the conversion efficiency of solar energy to H(2) also are discussed.     

Extraction of raw sewage sludge containing iron phosphate for phosphorus recovery

The objective of the present study was to establish an alkali extraction technology for FePO₄-containing sewage sludge obtained from a wastewater treatment system that includes phosphorous removal by iron electrolysis. By clarifying the extraction properties of phosphorous, organic matter, and inorganic matter, conditions for alkali extraction were optimized. As a result, it was suggested that unheated phosphorous extraction would be superior for FePO₄-containing sewage sludge. And, extraction methods and sewage sludge properties were also compared, and the noteworthy result that extraction of metals can be suppressed to extremely low amounts with alkali extraction as compared with acid extraction was obtained. A new insight was also gained that, as compared with the use of incinerated ash reported in previous studies, alkali extraction was more efficient when raw sewage sludge was used.     

Effects of tributyltin on the energy metabolism of pen shell (Atrina pectinata japonica)

We examined the effects of tributyltin (TBT) on aerobic and anaerobic energy metabolism of pen shell (Atrina pectinata japonica). We exposed pen shells to TBT at nominal concentrations of 0 (control) and 1.0microg/l for 72h under aerobic condition. At the end of the exposure, half of the pen shells in each treatment were wrapped in plastic wrap to simulate exposure to hypoxia and held at 25 degrees C for another 12h. The concentrations of the products of energy metabolism, namely lactate, pyruvate, fumarate and succinate, in adductor muscle were measured. The exposure to TBT under aerobic condition significantly elevated lactate, pyruvate and fumarate concentrations (p<0.001). After subsequent exposure to anaerobic condition, the mean concentration of succinate in the TBT treatment group was 64% of that in the control group, but there were no significant differences. Our results suggest that the energy metabolism of pen shell is disrupted by exposure to TBT.     

Historical and geographical aspects of the increasing perfluorooctanoate and perfluorooctane sulfonate contamination in human serum in Japan

Perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) have recently received attention due to their widespread contamination in the environment, as well as in wildlife and humans. We measured the PFOS and PFOA concentrations in historically recorded human serum samples at an age range between 20 and 59 years collected in Kyoto, 20 persons per each time point (n=100), and also the PFOS and PFOA concentrations in human serum samples at an age range between 20 and 59 years from 10 locations throughout Japan (n=200). The historical samples collected from 1983 to 1999 demonstrated that the PFOA concentrations in males and females from Kyoto have increased 4.4-fold and 4.3-fold at a rate of increase of 0.49 ng/ml/year and 0.42 ng/ml/year, respectively. In contrast, serum concentrations of PFOS reached a plateau in the late 1980s. There are also regional differences in both the PFOS and PFOA serum concentrations. The concentrations in serum [geometric mean (geometric standard deviation)] (ng/ml) in 2003-2004 ranged from 7.6(1.6) in the town of Matsuoka in Fukui prefecture to 27.8(1.6) in Kyoto city, and ranged from 2.3(1.5) in Matsuoka to 14.5(1.3) in Osaka city for PFOS and PFOA, respectively.     

Impact of solids on biphasic biodegradation of phenanthrene in the presence of hydroxypropyl-<Emphasis Type="Italic">β</Emphasis>-cyclodextrin (HPCD)

The consequence of polycyclic aromatic hydrocarbons (PAHs) in the environment is of great concern. The hydrophobic properties of PAHs significantly impact phase distribution causing limited bioavailability. Enhanced biodegradation has been extensively carried out by surfactants and the redeployment effect was recognized. However, the quantitative relationship concerning the impact of solids was rarely reported. A batch of biphasic tests were carried out by introducing Mycobacterium vanbaalenii PYR-1 and hydroxypropyl-β-cyclodextrin (HPCD) into a mixture of phenanthrene solution and various glass beads (GB37-63, GB105-125, and GB350-500). The comparative results demonstrated that HPCD had little effect on microbial growth and was not degradable by bacterium. A model was proposed to describe the biodegradation process. The regression results indicated that the partition coefficient k _d (1.234, 0.726 and 0.448 L·g^?1) and the degradation rate k (0 mmol·L^?1: 0.055, 0.094, and 0.112; 20 mmol·L^?1: 0.126, 0.141, and 0.156; 40 mmol·L^?1: 0.141, 0.156 and 0.184 d^?1) were positively and negatively correlated with the calculated total surface area (TSA) of solids, respectively. Degradation enhanced in the presence of HPCD, and the enhancing factor f was calculated (20 mmol·L^?1: 15.16, 40.01, and 145.5; 40 mmol·L^?1: 13.29, 37.97, and 138.4), indicating that the impact of solids was significant for the enhancement of biodegradation.     

Ocean acidification reduces biomineralization-related gene expression in the sea urchin, Hemicentrotus pulcherrimus

Although recent studies have demonstrated that calcification in a wide range of marine organisms is profoundly affected by CO₂-induced ocean acidification, the mechanism of this phenomenon is still unclear. To clarify the effects of ocean acidification on the calcification process at the molecular level, we evaluated the expression of three biomineralization-related genes in the sea urchin Hemicentrotus pulcherrimus exposed under control, 1,000, and 2,000?ppm CO₂ from egg to pluteus larval stage. We found that the expression of the gene msp130, which is proposed to transport Ca²⁺ to the calcification site, is suppressed by increased CO₂ at pluteus larval stage. Meanwhile, expression of the spicule protein matrix genes SM30 and SM50 was apparently not affected. The results suggest that the combined effects of ocean acidification on the expression of skeletogenesis-related genes as well as the change in seawater carbonate chemistry affect the biomineralization ability of sea urchins.     

An Integrated Assessment by Models for Energy Systems Analysis and Life-Cycle Assessment with a Case Study of Advanced Fossil-Fired Power Plants in China

We developed an integrated assessment (IA) using models for energy systems analysis and life-cycle assessment (LCA). Based on this assessment framework, we developed cost-benefit analysis (CBA) case studies for a hypothetical project designed to introduce advanced fossil-fired power generation technologies in China. Our MARKAL model for Japan confirmed that radical reductions (i.e., 80 % by 2050) of carbon dioxide (CO₂) could be attained from energy systems alone and that credit for emission allowances was required. We evaluated life-cycle costs and emissions of carbon dioxide, sulfur oxide, and nitrogen oxide gases for the energy technologies using an LCA model. Further, we applied a power generation planning model for six Chinese grids to provide a power mix structure, potentially producing credit by installing fossil-fired power generation technology and by using baseline grid emission factors with an average cost of electricity. Finally, by using dynamic emission reductions and additional costs from the two models, we conducted case studies of CBA for a hypothetical project to install the technologies in China. This was accomplished by evaluating emission reductions in monetary terms and by applying a life-cycle impact assessment model. A unique feature of our IA is its dynamic (time-varying) assessment of costs and benefits.     

Analysis of biomagnification of persistent organic pollutants in the aquatic food web of the Mekong Delta, South Vietnam using stable carbon and nitrogen isotopes

The present study elucidated the biomagnification profiles of persistent organic pollutants (POPs) through a tropical aquatic food web of Vietnam based on trophic characterization using stable nitrogen analysis. Various biological samples collected from the main stream of the Mekong Delta were provided for the analysis for both POPs, and stable nitrogen and carbon isotope ratios. Of the POPs analyzed, dichlorodiphenyltrichloroethane and its metabolites (DDTs) were the predominant contaminants with concentrations ranging from 0.058 to 12 ng/g wet weight, followed by polychlorinated biphenyls (PCBs) at 0.017-8.9 ng/g, chlordane compounds (CHLs) at 0.0043-0.76 ng/g, tris-4-chlorophenyl methane (TCPMe) at N.D.-0.26 ng/g, hexachlorocyclohexane isomers (HCHs) at N.D.-0.20 ng/g and hexachlorobenzene (HCB) at 0.0021-0.096 ng/g. Significant positive increases of concentrations in DDTs, CHLs, and TCPMe against the stable nitrogen ratio (delta(15)N) were detected, while, concentrations of HCHs and HCB showed no significant increase. The slopes of the regression equations between the log-transformed concentrations of these POPs and delta(15)N were used as indices of biomagnification. The slopes of the POPs for which positive biomagnification was detected ranged from 0.149 to 0.177 on a wet weight basis. The slopes of DDTs and CHLs were less than those reported for a marine food web of the Arctic Ocean, indicating that less biomagnification had occurred in the tropical food web. Of the isomers of CHLs, unlike the studies of the Arctic Ocean, oxychlordane did not undergo significant biomagnification through the food web of the Mekong Delta. This difference is considered to be due to a lack of marine mammals, which might metabolize cis- and trans-chlordane to oxychlordane, in the Mekong Delta ecosystem. The biomagnification profile of TCPMe is reported for the first time in the present study.     

Carbon black nanoparticles promote the maturation and function of mouse bone marrow-derived dendritic cells

Particulate matter including carbon black (CB) nanoparticles can enhance antigen-related inflammation and immunoglobulin production in vivo. Dendritic cells (DC) as antigen-presenting cells (APC) are the most capable inducers of immune responses. The present study was designed to determine whether CB nanoparticles affect the maturation/activation and function of DC in vitro. DC were differentiated from bone marrow (BM) cells of BALB/c mice by culture with granulocyte macrophage colony stimulating factor (GM-CSF). At day 8 of culture, BM-derived DC (BMDC) were exposed to CB nanoparticles with a diameter of 14nm or 56nm for 24h. The expression of major histocompatibility complex (MHC) class II, DEC205, CD80, and CD86 (maturation/activation markers of BMDC) was measured by flow cytometry. BMDC function was evaluated by an allogeneic mixed lymphocyte reaction (MLR) assay. CB nanoparticles significantly increased the expression of DEC205 and CD86 in BMDC and tended to increase MHC class II and CD80 expression; however, a size-dependent effect was not observed. On the other hand, BMDC-mediated MLR was significantly enhanced by the CB nanoparticles and the enhancement was greater by 14nm CB nanoparticles than by 56nm CB nanoparticles. Taken together, CB nanoparticles can promote the maturation/activation and function of BMDC, which could be related to their effects on allergic diseases and/or responses. In addition, BMDC-mediated MLR might be useful assay for in vitro screening for adjuvant activity of environmental toxicants.     

Formation of N-nitrosodimethylamine (NDMA) by ozonation of dyes and related compounds

Formation of N-nitrosodimethylamine (NDMA) by ozonation of commercially available dyes and related compounds was investigated. Ozonation was conducted using a semi-batch type reactor, and ozone concentration in gas phase and the ozone gas flow were 10 mg L(-1) and 1.0 L min(-1), respectively. NDMA was formed by 15 min of ozonation of seven out of eight selected target compounds (0.05 mM) at pH 7. All the target compounds with N,N-dimethylamino functions were NDMA precursors in ozonation. The lowest and highest NDMA concentrations after ozonation of the target compounds were 13 ng L(-1) for N,N-dimethylformamide (DMF) and 1600 ng L(-1) for N,N-dimethyl-p-phenylenediamine (DMPD), respectively. NDMA concentrations after 15 min of ozonation of 0.05 mM methylene blue (MB) and DMPD increased with an increase in pH in its range of 6-8. The effects of coexisting compounds on NDMA concentrations after 15 min of ozonation of 0.05 mM MB and DMPD were examined at pH 7. NDMA concentrations after ozonation of MB and DMPD increased by the presence of 0.05 mM (0.7 mg L(-1) as N) nitrite (NO(2)(-)); 5000 ng L(-1) for MB and 4000 ng L(-1) for DMPD. NDMA concentration after MB ozonation decreased by the presence of 5mM tertiary butyl alcohol (TBA), a hydroxyl radical (HO.) scavenger, but that after DMPD ozonation was increased by the presence of TBA. NDMA concentrations after ozonation of MB and DMPD were not affected by the presence of 0.16 mM (5.3 mg L(-1)) hydrogen peroxide (H(2)O(2)). When 0.05 mM MB and DMPD were added to the Yodo and Tone river water samples, NDMA concentrations after 15 min of their ozonation at pH 7 increased compared with those in the case of addition to ultrapure water samples.