Genotoxicity of nano/microparticles in in vitro micronuclei, in vivo comet and mutation assay systems

Background

Recently, manufactured nano/microparticles such as fullerenes (C₆₀), carbon black (CB) and ceramic fiber are being widely used because of their desirable properties in industrial, medical and cosmetic fields. However, there are few data on these particles in mammalian mutagenesis and carcinogenesis. To examine genotoxic effects by C₆₀, CB and kaolin, an in vitro micronuclei (MN) test was conducted with human lung cancer cell line, A549 cells. In addition, DNA damage and mutations were analyzed by in vivo assay systems using male C57BL/6J or gpt delta transgenic mice which were intratracheally instilled with single or multiple doses of 0.2 mg per animal of particles.

Results

In in vitro genotoxic analysis, increased MN frequencies were observed in A549 cells treated with C₆₀, CB and kaolin in a dose-dependent manner. These three nano/microparticles also induced DNA damage in the lungs of C57BL/6J mice measured by comet assay. Moreover, single or multiple instillations of C₆₀ and kaolin, increased either or both of gpt and Spi^- mutant frequencies in the lungs of gpt delta transgenic mice. Mutation spectra analysis showed transversions were predominant, and more than 60% of the base substitutions occurred at G:C base pairs in the gpt genes. The G:C to C:G transversion was commonly increased by these particle instillations.

Conclusion

Manufactured nano/microparticles, CB, C₆₀ and kaolin, were shown to be genotoxic in in vitro and in vivo assay systems.     

Effects of rice straw returning methods on N<Subscript>2</Subscript>O emission during wheat-growing season

A field experiment was conducted in Jurong of Nanjing, Jiangsu Province, China from 2006 to 2008 to investigate N₂O emission during the wheat-growing season as affected by various rice straw returning methods prior to wheat cultivation. The study was designed to have four treatments: no rice straw applied (CK), rice straw burnt in situ (RB), rice straw evenly incorporated into the topsoil (RI), rice straw evenly spread over the field as mulch (RM). Results showed that N₂O emission was decreased by 24–29% in Treatment RB and by 3–18% in Treatment RI, but increased by 15–39% in Treatment RM, compared with that in Treatment CK. The contents of soil total C and N at wheat harvest were significantly increased by 7–13% and by 8–12% in Treatment RI, respectively, compared with that in Treatment CK. The wheat grain yield in Treatment RI was 1.0–1.2 times that in the Treatment CK. Based on these results, the best management practice of returning rice straw to the soil prior to wheat cultivation is evenly incorporating rice straw into the topsoil, as the method tended to reduce N₂O emission during the wheat-growing season and increase wheat yield and soil fertility.     

Stabilization of flavylium dye by incorporation into the pore of protonated zeolites

Flavylium dye, a model compound of natural anthocyanin, has a defect that its color stability is very poor. We have complexed the flavylium dye with protonated zeolites and succeeded to enhance both the thermal and chemical stability of the dye. The aqueous dispersion of the flavylium/zeolite complex maintains its color even at 353 K or at pH 9, under which conditions the aqueous solution of the flavylium loses its color immediately. The stability is enhanced when the dye is incorporated into the pore of the zeolites, and a large stabilization effect is obtained by a tight adsorption with a narrow pore, as long as the dye is incorporated into the pore. Because of its enhanced color stability, the flavylium dye incorporated in the zeolite pore can be a candidate for an environmentally friendly coloring material.     

Effects of amino acid replacements around the reactive site of chicken ovomucoid domain 3 on the inhibitory activity toward chymotrypsin and trypsin

We have previously shown that replacing the P1-site residue(Ala) of chicken ovomucoid domain 3 (OMCHI3) with a Met or Lysresults in the acquisition of inhibitory activity toward chymotrypsinor trypsin, respectively. However, the inhibitory activitiesthus induced are not strong. In the present study, we introducedadditional amino acid replacements around the reactive siteto try to make the P1-site mutants more effective inhibitorsof chymotrypsin or trypsin. The amino acid replacement AspTyrat the P2' site of OMCHI3(P1Met) resulted in conversion to a35000-fold more effective inhibitor of chymotrypsin with aninhibitor constant (K_i) of 1.17x10^–11 M. The K_i valueof OMCHI3(P1Met, P2'Ala) indicated that the effect on the interactionwith chymotrypsin of removing a negative charge from the P2'site was greater than that of introducing an aromatic ring.Similarly, enhanced inhibition of trypsin was observed whenthe AspTyr replacement was introduced into the P2' site of OMCHI3(P1Lys).Two additional replacements, AspAla at the P4 site and ArgAlaat the P3' site, made the mutant a more effective inhibitorof trypsin with a K_i value of 1.44x10^–9 M. By contrast,ArgAla replacement at the P3' site of OMCHI3(P1Met, P2'Tyr)resulted in a greatly reduced inhibition of chymotrypsin, andAspAla replacement at the P4 site produced only a small changewhen compared with a natural variant of OMCHI3. These resultsclearly indicate that not only the P1-site residue but alsothe characteristics, particularly the electrostatic properties,of the amino acid residues around the reactive site of the proteaseinhibitor determine the strength of its interactions with proteases.Furthermore, amino acids with different characteristics arerequired around the reactive site for strong inhibition of chymotrypsinand trypsin.     

CO2 hydrogenation over micro‐ and mesoporous oxides supported Ru catalysts

We prepared relatively uniform supported Ru catalysts by ion‐exchange and CVD methods, using an NaY zeolite and a mesoporous FSM‐16 as substrates, and carried out CO₂ hydrogenation. They showed high activity for CO₂ hydrogenation. A Ru‐ion‐exchanged catalyst showed high activity for methanol production. Co addition promoted methanol formation. This revised version was published online in July 2006 with corrections to the Cover Date.     

Thermal conduction theoretical analysis of temperature distribution during multiple-electrode submerged arc welding

In recent years, in order to reduce the costs of transportation and construction of pipelines, which are often constructed using multiple-electrode submerged arc welding (SAW), higher joint performance is required. Therefore, there has arisen the need to understand theoretically and control appropriately metallurgical and mechanical characteristics in heat-affected zone (HAZ), which has a significant influence on the strength and toughness of welded joints. Commonly, metallurgical phenomena in HAZ are evaluated based on the highest temperature and the cooling rate. Therefore, in order to control metallurgical and mechanical characteristics in HAZ by means of the welding conditions, evaluating the temperature distribution and the temperature history near the melted zone is essential. However, a detailed investigation of the temperature distribution for multiple-electrode SAW has not yet been carried out enough. In this study, in order to investigate the temperature distribution and histories during multiple-electrode SAW, the experimental results are compared with the theoretical results. In the theoretical analysis, the temperature rise equation in multiple heat sources welding is developed using the method of summation. Furthermore, on temperature distribution during welding, the effects of multiple heat sources, such as the number of heat sources and the distance between each electrodes, are considered quantitatively through the thermal conduction theoretical analysis. As the result, the distance between lead heat source and final heat source primally influences the area with the difference between a single heat source welding and multiple heat sources welding. Based on the results, it is expected to control temperature distribution near melted zone by more appropriate heat input characteristics, which is depended on heat source arrangement.     

MiR-10a in Pancreatic Juice as a Biomarker for Invasive Intraductal Papillary Mucinous Neoplasm by miRNA Sequencing

New biomarkers are needed to further stratify the risk of malignancy in intraductal papillary mucinous neoplasm (IPMN). Although microRNAs (miRNAs) are expected to be stable biomarkers, they can vary owing to a lack of definite internal controls. To identify universal biomarkers for invasive IPMN, we performed miRNA sequencing using tumor-normal paired samples. A total of 19 resected tissues and 13 pancreatic juice samples from 32 IPMN patients were analyzed for miRNA expression by next-generation sequencing with a two-step normalization of miRNA sequence data. The miRNAs involved in IPMN associated with invasive carcinoma were identified from this tissue analysis and further verified with the pancreatic juice samples. From the tumor-normal paired tissue analysis of the expression levels of 2792 miRNAs, 20 upregulated and 17 downregulated miRNAs were identified. In IPMN associated with invasive carcinoma (INV), miR-10a-5p and miR-221-3p were upregulated and miR-148a-3p was downregulated when compared with noninvasive IPMN. When these findings were further validated with pancreatic juice samples, miR-10a-5p was found to be elevated in INV (p = 0.002). Therefore, three differentially expressed miRNAs were identified in tissues with INV, and the expression of miR-10a-5p was also elevated in pancreatic juice samples with INV. MiR-10a-5p is a promising additional biomarker for invasive IPMN.     

Supramolecular Nanofibers from Collagen-Mimetic Peptides Bearing Various Aromatic Groups at N-Termini via Hierarchical Self-Assembly

Self-assembly of artificial peptides has been widely studied for constructing nanostructured materials, with numerous potential applications in the nanobiotechnology field. Herein, we report the synthesis and hierarchical self-assembly of collagen-mimetic peptides (CMPs) bearing various aromatic groups at the N-termini, including 2-naphthyl, 1-naphtyl, anthracenyl, and pyrenyl groups, into nanofibers. The CMPs (R-(GPO)_n: n > 4) formed a triple helix structure in water at 4 °C, as confirmed via CD analyses, and their conformations were more stable with increasing hydrophobicity of the terminal aromatic group and peptide chain length. The resulting pre-organized triple helical CMPs showed diverse self-assembly into highly ordered nanofibers, reflecting their slight differences in hydrophobic/hydrophilic balance and configuration of aromatic templates. TEM analysis demonstrated that 2Np-CMP_n (n = 6 and 7) and Py-CMP₆ provided well-developed natural collagen-like nanofibers and An-CMP_n (n = 5–7) self-assembled into rod-like micelle fibers. On the other hand, 2Np-CMP₅ and 1Np-CMP₆ were unable to form nanofibers under the same conditions. Furthermore, the Py-CMP₆ nanofiber was found to encapsulate a guest hydrophobic molecule, Nile red, and exhibited unique emission behavior based on the specific nanostructure. In addition to the ability of CMPs to bind small molecules, their controlled self-assembly enables their versatile utilization in drug delivery and wavelength-conversion nanomaterials.     

Kinetics of CO2 reforming of methane by catalytically activated metallic foam absorber for solar receiver-reactors

Ni–Cr–Al metallic foam absorber with high porosity was catalytically activated using a Ru/γ-Al₂O₃ catalyst, and was subsequently tested with respect to CO₂ reforming of methane in a small-scale volumetric receiver-reactor by using a sun-simulator. A chemical storage efficiency of 37% was obtained for a mean light flux of 325 kW m⁻². Furthermore, the activity and the stability of the metallic foam absorber were compared with those of a SiC foam absorber activated with the same Ru/γ-Al₂O₃ catalyst for 50 h of light irradiation, and it was found that the metallic foam absorber has superior catalytic stability in comparison to the SiC form absorber. In addition, unlike ceramic foams such as SiC, metallic foams feature superior plasticity, which prevents the emergence of cracks caused by mechanical or thermal shock.     

ZnO nano-ridge structure and its application in inverted polymer solar cell

We report a unique nano-ridge structure of zinc oxide (ZnO) and its application in high performance inverted polymer solar cells. The ZnO nano-ridge structure was formed by a sol–gel process using a ramp annealing method. As the solvent slowly evaporated due to the low heating rate, there was sufficient time for the gel particles to structurally relax and pile up, resulting in a dense and undulated film. Nano-ridges with peak as high as 120 nm and valley to valley distance of about 500 nm were formed. This film provided an effective hole blocking layer and also an increased interfacial area for electron collection. An inverted bulk heterojunction polymer solar cell was fabricated using the ZnO nano-ridge film as the electron collecting layer. The device showed a high power conversion efficiency of 4.00%, an improvement of about 25% over similar solar cells made with a planar film of ZnO nanoparticles.