MicroRNA-331-3p Suppresses Cervical Cancer Cell Proliferation and E6/E7 Expression by Targeting NRP2

Aberrant expression of microRNAs (miRNAs) is involved in the development and progression of various types of cancers. In this study, we investigated the role of miR-331-3p in cell proliferation and the expression of keratinocyte differentiation markers of uterine cervical cancer cells. Moreover, we evaluated whether neuropilin 2 (NRP2) are putative target molecules that regulate the human papillomavirus (HPV) related oncoproteins E6 and E7. Cell proliferation in the human cervical cancer cell lines SKG-II, HCS-2, and HeLa was assessed using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. Cellular apoptosis was measured using the TdT-mediated dUTP nick end labeling (TUNEL) and Annexin V assays. Quantitative RT-PCR was used to measure the messenger RNA (mRNA) expression of the NRP2, E6, E7, p63, and involucrin (IVL) genes. A functional assay for cell growth was performed using cell cycle analyses. Overexpression of miR-331-3p inhibited cell proliferation, and induced G2/M phase arrest and apoptosis in SKG-II, HCS-2 and HeLa cells. The luciferase reporter assay of the NRP2 3′-untranslated region revealed the direct regulation of NRP2 by miR-331-3p. Gene expression analyses using quantitative RT-PCR in SKG-II, HCS-2, and HeLa cells overexpressing miR-331-3p or suppressing NRP2 revealed down-regulation of E6, E7, and p63 mRNA and up-regulation of IVL mRNA. Moreover, miR-331-3p overexpression was suppressed NRP2 expression in protein level. We showed that miR-331-3p and NRP2 were key effectors of cell proliferation by regulating the cell cycle, apoptosis. NRP-2 also regulates the expression of E6/E7 and keratinocyte differentiation markers. Our findings suggest that miR-331-3p has an important role in regulating cervical cancer cell proliferation, and that miR-331-3p may contribute to keratinocyte differentiation through NRP2 suppression. miR-331-3p and NRP2 may contribute to anti-cancer effects.     

Porous Titania Ceramic Prepared by Mimicking Silicified Wood

A porous titania ceramic with a woodlike microstructure that was analogous to silicified wood was prepared. The production of "titanified wood" was performed using the following process: (i) introduction of titanium tetraisopropoxide into wood materials via vacuum infiltration, (ii) hydrolysis of the titanium tetraisopropoxide in the cell structure to form a titania gel, and (iii) firing at a temperature of 600°–1400°C in air. The resulting titania ceramic had the same external and internal forms of the original wood.     

Vibrationally Excited CO2 Formed in CO + NO Reaction on Pd(110) Surface in High Surface Temperature Range

The infrared chemiluminescence spectra of CO₂ formed during steady-state CO+NO reaction over Pd(110) indicated that the temperature of the bending vibrational mode was much higher than that of the antisymmetric one at higher surface temperatures such as 800–850 K. Especially, in the high temperature range, more vibrationally excited CO₂ was formed from CO+NO reaction than CO+O₂ reaction. On the basis of the result, we propose the model structure of reaction intermediates for CO₂ formation in CO+NO reaction, which is different from that in CO+O₂ reaction.     

Effect of Lipase Activity and Specificity on the DAG Content of Olive Oil from the Shodoshima-Produced Olive Fruits

Olive oils have a higher relative diacylglycerol (DAG) content than other plant oils. The lipase in olive fruits is involved in DAG production and is directly related to the acidity of the olive oil. However, the lipase activity and positional selectivity have not been clarified. To investigate the properties of olive fruit lipase, olive fruits of the Mission variety harvested during mid-December of 2005 on Shodoshima Island (Japan) were stored at 20, 30 or 40 °C for 4 weeks. Changes in the acidity and acylglycerol content of the oils extracted from the stored fruits were analyzed. The acidity and DAG content of the olive oils increased due to triacylglycerol (TAG) hydrolysis during storage. sn-1,2-DAGs preferentially increased during the early stages of storage, indicating that the olive fruit lipase is enantioselective for the sn-3 position, while non-enzymatic isomerization of sn-1,2-DAGs was observed throughout the entire duration of storage. Kinetic analysis revealed that the enantioselectivity of olive fruit lipase for the sn-3 position was approximately four times greater than for the sn-1 position. The lipase was gradually inactivated at temperatures of 30 °C or higher, and the ratios of the rate constant for inactivation to TAG hydrolysis at the sn-3 position was 0.2, 13, and 23 at 20, 30, and 40 °C, respectively.     

On-line measurement of orientation development in the high-speed melt spinning process

On-line measurement of birefringence was performed in the high-speed melt spinning process of poly(ethylene terephthalate) using an apparatus that incorporates a rotating polarizer for the measurement of the optical retardation of running filament. Particular attention was paid to the detailed measurements in the vicinity of neck-like deformation. Through the measurement at the take-up velocity of 5 km/min, development of birefringence under the strain rate up to about 1 ms^?1 was investigated. To analyze the relation between applied stress and birefringence, tension and temperature profiles of the spin-line were calculated based on the experimentally obtained diameter profiles. Even though the strain rate is extremely high, a linear relationship between birefringence and a parameter calculated by dividing stress by temperature was confirmed to hold up to birefringence and stress/temperature values of about 0.017 and 10 kPa/K, respectively.     

Detection of tris(2,4-di-tert-butylphenyl)phosphite (Irgafos 168, antioxidant in plastics) and its oxide in commercial frozen vegetables

Two peaks, A and B, detected in chromatograms of commercial frozen vegetable extracts during analysis of organophosphorus pesticide residues by GC-FPD, were identified as tris(2,4-ditert-butylphenyl)phosphite (Irgafos 168) and Irgafos 168 oxide, respectively, from their mass spectra. Irgafos 168 is used as an antioxidant in plastics, and there has been no report of its detection in foods. We analyzed Irgafos 168 and its oxide in 38 samples of commercial frozen vegetables, and they were detected from 4 samples (0.02-0.80 microgram/g as total amount of Irgafos 168 and its oxide).     

Pseudomonas aeruginosa as a model microorganism for investigation of chemotactic behaviors in ecosystem

Motile bacteria sense changes in the concentration of chemicals in environments and respond in a behavioral manner. This behavioral response is called chemotaxis. Bacterial chemotaxis can be viewed as an important prelude to metabolism, prey-predator relationships, symbiosis, infections, and other ecological interactions in biological communities. Genome analysis reveals that a large number of environmental motile bacteria possess a number of genes involved in chemosensing and chemotatic signal transduction. Pseudomonas aeruginosa has a very complex chemosensory system with more than 20 chemotaxis (che) genes in five distinct clusters and 26 chemoreceptor (methyl-accepting chemotaxis protein [mcp]) genes. Among the 26 MCPs of P. aeruginosa, nine have been identified as MCPs for amino acids, inorganic phosphate, oxygen, ethylene, and volatile chlorinated aliphatic hydrocarbons, whereas 3 MCPs were demonstrated to be involved in biofilm formation and biosynthesis of type IV pilus. Six che genes are essential for chemotactic responses, while genes in Pil-Chp cluster and Wsp cluster are involved in type IV pilus synthesis and twitching motility and biofilm formation, respectively. P. aeruginosa, with its complex chemotaxis system, is a better model microorganism for investigating ecological aspects of chemotaxis in environmental bacteria than Escherichia coli and Salmonella enterica serovar Typhimurium, which possess a relatively simpler chemotaxis system.     

Preparation of detergent-lipase complexes utilizing water-soluble amphiphiles in single aqueous phase and catalysis of transesterifications in homogeneous organic solvents

A novel method of preparing detergent-enzyme complexes that can be employed in organic media was developed utilizing newly synthesized water-soluble nonionic gemini-type detergents, N,N-bis(3-D-gluconamidopropyl)-3-(dialkyl-L-glutamatecarbonyl)propanamides (BIG2CnCA: n = 10,12,14,16,18) and N,N-bis(3-D-lactonamidopropyl)-3-(dialkyl-L-glutamatecarbonyl)propanamides (BIL2CnCA: n = 16,18), and nonionic twin-headed detergents, N,N-bis(3-D-gluconamidopropyl)alkanamides (BIG1Cn: n = 12,14,16,18,delta9). This method simply entails mixing a selected enzyme with an appropriate detergent in an aqueous solution followed by lyophilization, and it offers the advantages of enhanced enzymatic activity in organic solvents and eliminates both enzyme loss and the necessity for an organic solvent in the preparation stage. Using various modified lipases originating from Aspergillus niger (Lipase A), Candida rugosa (Lipase C), Pseudomonas cepacia (Lipase P), and porcine pancreas (PPL), prepared using the novel method and detergents, including conventional synthesized nonionic detergents such as dialkyl N-D-glucona-L-glutamates (2CnGE: n = 12,18delta9) and octanoyl-N-methylglucamide (MEGA-8), enantioselective transesterifications of 6-methyl-5-hepten-2-ol (sulcatol) and 2,2-dimethyl-1,3-dioxolane-4-methanol (solketal) with a vinyl or isopropenyl carboxylate were carried out in an organic solvent. The modified lipase activity was influenced by both the lipases and the structure of the detergents. The value for the hydrophile-lipophile balance (HLB) of the detergent provided a means of correlating the structure and the obtained modified lipase activity. For detergents of the same class with a HLB value of approximately 9 and 12, the highest activity was obtained for Lipase A and Lipase P, and Lipase C and PPL, respectively. Among detergents of the same HLB value tested, the gemini-type detergents possessing the most bulky head and tail were most effective as a modifier for lipases of all types. The preparation and reaction conditions for these novel gemini-type detergent-modified lipases were optimized using BIG2C12CA (HLB = 9.4) by studying the effect of the detergent/lipase ratio and the nature of organic solvents on the complex formation. The high enzymatic activities of the BIG2C12CA-modified lipases were independent of the solubility of the lipases in organic solvents, unlike in the case of 2CnGE-modified lipases prepared using the conventional suspension system.