Gene Expression over Time during Cell Transformation Due to Non-Genotoxic Carcinogen Treatment of Bhas 42 Cells

The Bhas 42 cell transformation assay (Bhas 42 CTA) is the first Organization for Economic Cooperation and Development (OECD)-certificated method used as a specific tool for the detection of the cell-transformation potential of tumor-promoting compounds, including non-genotoxic carcinogens (NGTxCs), as separate from genotoxic carcinogens. This assay offers the great advantage of enabling the phenotypic detection of oncotransformation. A key benefit of using the Bhas 42 CTA in the study of the cell-transformation mechanisms of tumor-promoting compounds, including non-genotoxic carcinogens, is that the cell-transformation potential of the chemical can be detected directly without treatment with a tumor-initiating compound since Bhas 42 cell line was established by transfecting the v-Ha-ras gene into a mouse fibroblast cloned cell line. Here, we analyzed the gene expression over time, using DNA microarrays, in Bhas 42 cells treated with the tumor-promoting compound 12-O-tetradecanoylphorbol-13-acetate (TPA), and NGTxC, with a total of three repeat experiments. This is the first paper to report on gene expression over time during the process of cell transformation with only a tumor-promoting compound. Pathways that were activated or inactivated during the process of cell transformation in the Bhas 42 cells treated with TPA were related not only directly to RAS but also to various pathways in the hallmarks of cancer.     

Purification and characterization of ouabain-binding protein in human plasma

Ouabainlike factors are thought to be a kind of important modulators of salt and water metabolism in essential hypertension. We purified the binding-protein of ouabain (OBP) from human plasma. The amino-terminal sequence of OBP from human plasma, (NH2-TLGQPREPQVYTLPPXREEM-), indicated that OBP is the carboxy-terminal fragment (14.4 kDa by SDS-PAGE) from T218 of IgG2 heavy chain and from A221 of the IgG1 heavy chain constant region. Moreover, plasmin-cleaved Fc fragment (pFc) of IgG possessed the ouabain-binding activity by the gel-filtration method of pFc and authentic ouabain mixture, whereas neither intact, aggregate, nor papain-cleaved Fc fragment did. The amino-terminal sequence of pFc was NH2-THTXPPXPAPELLGGPXVFL-, and this sequence corresponded to the T105 to L125 fragment of the IgG1 heavy chain constant region. The growth of cultured THP-1 cells were arrested in the dose-dependent manner by ouabain, which was inhibited by the addition of 20 microg/mL of pFc. These results suggested that plasmin-cleaved Fc of human IgG is one of the binding protein of ouabain/ouabainlike factor(s) in human plasma.     

Enhanced renal toxicity by inorganic mercury in metallothionein-null mice

To elucidate a protective role of metallothionein (MT) in the manifestation of inorganic mercury toxicity, we studied the susceptibility of MT-null mice to the renal toxicity of mercuric chloride. Because the MT-null (J) mice are a genetic background of 129/Sv strain, the 129/Sv mice were used as wild-type controls. Nine-week-old male MT-null (J) and 129/Sv mice were given subcutaneous injections of mercuric chloride at doses of 10 to 40 micromol/kg. The basal MT level in the kidney of MT-null (J) mice was undetectable (<0.2 microg/g of tissue) and approximately 2.5 microg/g of tissue in 129/Sv mice. The sensitivity to the renal toxicity of mercuric chloride was markedly enhanced in the MT-null (J) mice compared with the 129/Sv mice. The renal mercury level was similar for the MT-null (J) and 129/Sv mice at 4 hr after the injection of mercuric chloride (20 micromol/kg) but became significantly lower in MT-null (J) mice than in 129/Sv mice at 24 and 72 hr. Based on the present results, we conclude that MT is an important protective factor against the renal toxicity caused by inorganic mercury and that it may play a major role in the retention of mercury in the kidney.     

Carbonization of aromatic hydrocarbons—V: Microscopic features of carbons obtained by the aid of catalysts

Microscopic observation of carbons obtained from pure aromatic hydrocarbons by the aid of carbonization catalysts was carried out to clarify the microstructure of these carbons of different features. Reflected polarized-light microscopy distinguished needle, mosaic and isotropic cokes, former two of which were produced with aluminum chloride and the last with potassium. High resolution microscopy revealed that these carbons calcined at 1250° had different degree of layered structure, corresponding to the crystallographic parameters of these samples graphitized at 2500°C. The reasons for the carbons produced with potassium to be non-graphitizable are discussed from the macro- and micro-features of the carbons.     

Modifying carbonization properties of pitches. 3. Carbonization of modified quinoline-insoluble matter from pitches

The carbonization of solubilized matter obtained from the hydrogenated and reductively alkylated quinoline-insolubles of pitches was studied to clarify the different carbonization properties shown by these materials. Dehydrogenation of hydrogenated QI started at 200 °C but continued until 400 °C, passing through a fused phase to give graphitizable carbon. In contrast, alkylated QI gave non-graphitizable carbon because it readily reverted to QI by dealkylation below 300 °C, before fusion. QI alkylated with butyl or benzyl groups was found to be nearly 80% soluble in benzene.     

Studies on ion-exchange membranes. XXXII. Heterogeneity in ion-exchange membranes

Twelve kinds of cation-exchange membranes were treated with hydrogen peroxide. Some of them (Selemion CMV, Nepton CR-61, Scrion C-100, SAM-1) were completely destroyed. Heterogeneity is believed to be present in that part of their chemical structures that is decomposable by the treatment. The other membranes were converted into porous membranes by the decomposition of the resinous part. Water permeabilities and electric resistances of the porous membranes were examined to evaluate the pore radius. It was concluded that the “paste method” membranes have a heterogeneity or localized distribution of the resinous part under 50 mμ. The “paste method” membranes seemed to resemble the “graft method” membranes in chemical structure and to be much different from the mosaic-type membranes such as Permaplex and MC-3142. These results were also supported by extraction of the membranes with appropriate solvents and observation by electron microscopy.     

Allelopathy of yellow fieldcress (Rorippa sylvestris): Identification and characterization of phytotoxic constituents

Both the neutral and acidic fractions of the acetone extract of yellow fieldcress (Kireha-inugarashi,Rorippa sylvestris Besser) inhibited lettuce seed germination. Salicylic,p-hydroxybenzoic, vanillic, and syringic acid were identified in the acidic fraction. In the neutral fraction, hirsutin (8-methylsulfinyloctyl isothiocyanate), 4-methoxyindole-3-acetonitrile, and pyrocatechol were identified. Bioassay using a root exudate recirculating system showedR. sylvestris during flowering inhibited the lettuce seedling growth. Hirsutin (13g/plant/day) and pyrocatechol (9.3g/plant/day) were the major compounds released into the rhizosphere. Several combinations of pyrocatechol,p-hydroxybenzoic acid, vanillic acid, and hirsutin reduced lettuce seedling growth. These compounds seemed to be allelochemicals.     

Spider Mites Assess Predation Risk by Using the Odor of Injured Conspecifics

When a predatory mite, Amblyseius womersleyi is present, Tetranychus kanzawai takes refuge on its webs during a quiescent stage. To examine the factors responsible for the antipredator behavior of T. kanzawai, we exposed T. kanzawai to the odor of conspecifics, A. womersleyi, and both T. kanzawai and A. womersleyi. The proportion of T. kanzawai females that took refuge was significantly higher when they were exposed to the odor of `both T. kanzawai and A. womersleyi.' Furthermore, the proportion of T. kanzawai females that took refuge was significantly higher when they were exposed to injured conspecifics than to intact conspecifics. These results suggest that T. kanzawai assesses predation risk, at least in part, by using the odor of injured conspecifics.     

X-ray absorption fine structure study on residue bromine in carbons with different degrees of graphitization

The structure of bromine residue compounds was investigated by X-ray absorption fine structure (XAFS) in order to interpret where and how bromine is present in carbons with different degrees of graphitization. The residue compounds can be classified into three groups, as obtained from X-ray absorption near edge structure (XANES) spectra and the values of the intramolecular distance r_Br–Br determined by extended X-ray absorption fine structure (EXAFS). In Group I, prepared from the host carbons heat treated at temperatures higher than 1900 °C, bromine exists in the interlayer space of graphite in the form of Br₂ molecules with interaction of the π electrons of graphite. In Group III, from carbon heat treated at 1000 °C, most of the bromine probably reacts with carbon atoms having a dangling bond or functional groups. For Group II, where the host carbons are heat treated at intermediate temperatures, it is likely that bromine exists in undeveloped defects with a unique electronic state.     

Fracture behavior of core‐shell rubber–modified clay‐epoxy nanocomposites

Morphology and fracture mechanisms in two nanoclay‐filled epoxy systems were investigated using both microscopy and spectroscopy tools. Clay exfoliation was achieved using a series of sample preparation steps, and confirmed using wide angle X‐ray diffraction (XRD) and transmission electron microscopy (TEM) techniques. Significant improvement in modulus was obtained when clay exfoliation was achieved. Incorporation of core‐shell rubber (CSR) in both caly‐filled epoxy systems leads to greatly enhanced fracture toughness. Optical microscopy and TEM observations of the CSR‐modified nanocomposites suggest that CSR cavitation. shear yielding of the matrix, clay layer delamination. CSR bridging, crack bifurcation. and crack deflection are among the operative toughening mechanisms observed, depending on the nature of the epoxy matrix utilized.