Electrochemical DNA methylation detection for enzymatically digested CpG oligonucleotides

We describe the electrochemical detection of DNA methylation through the direct oxidation of both 5-methylcytosine (mC) and cytosine (C) in 5'-CG-3' sequence (CpG) oligonucleotides using a sputtered nanocarbon film electrode after digesting a longer CpG oligonucleotide with endonuclease P1. Direct electrochemistry of the longer CpG oligonucleotides was insufficient for obtaining the oxidation currents of these bases because the CG rich sequence inhibited the direct oxidation of each base in the longer CpG oligonucleotides, owing to the conformational structure and its very low diffusion coefficient. To detect C methylation with better quantitativity and sensitivity in the relatively long CpG oligonucleotides, we successfully used an endonuclease P1 to digest the target CpG oligonucleotide and yield an identical mononucleotide 2'-deoxyribonucleoside 5'-monophosphate (5'-dNMP). Compared with results obtained without P1 treatment, we achieved 4.4 times higher sensitivity and a wider concentration range for mC detection with a resolution capable of detecting a subtle methylated cytosine difference in the CpG oligonucleotides (60mer).     

Detection of microbubble position by a digital hologram

This paper reports on a new technique of measurements of microbubble position in three dimensions with high time-resolution. The technique is based on micro digital holographic particle tracking velocimetry. In this technique, an intensity profile is constructed from a holographic image of a microbubble where the profile results in showing two peaks. The distance between the two peaks appears to relate to the size of the microbubble's diameter. The three-dimensional position of the bubble can be detected by the center of the two peaks and the center point of the bubble image focused by a digital hologram. We also theoretically obtained the intensity profile of a microbubble by considering a refraction of light on a bubble surface to a ring-shaped aperture model. The theoretically obtained distance between the two peaks is found to be in good agreement with the values obtained experimentally.     

Study on the compressible viscous flows through a straight pipe

When sonic nozzles of significantly smaller diameter are used as standard flow meters, the critical backpressure ratio is affected by the boundary layer at the nozzle throat. It is known that, as the flow pathway is complicated in actual valves, the critical pressure ratio differs from the ideal value. However, the effect of the boundary layer thickness on choking criteria is still controversial. Choking phenomenon in a sonic nozzle fitted with a straight circular pipe of variable length is investigated numerically and the results are compared with experimental and theoretical ones. The results show that the actual critical pressure ratio is less than the ideal value and it decreases with increase in pipe length. The effect of boundary layer thickness on main flow Mach number is also made clear in this work. In addition, sonic conductance and pressure loss in actual viscous flows computed are compared with the ISO 6358 standard.     

Experimental study on hysteresis phenomena of shock wave structure in an over-expanded axisymmetric jet

In recent studies on two-dimensional supersonic jets, it is reported that the hysteresis phenomenon for the reflection type of shock waves in the jet flow field is occurred under the quasi-steady flow condition and this phenomenon is affected by the transitional pressure ratio between the regular reflection and Mach reflection. However, so far, there are few researches on the hysteresis phenomenon for the transition of shock waves between regular and Mach reflection in over-expanded supersonic jets and the phenomenon has not been investigated satisfactorily. Therefore, the purpose of this study is to clarify the hysteresis phenomena for the reflection type of shock wave in the over-expanded axi-symmetric supersonic jet experimentally, and to discuss the relationship between hysteresis phenomenon and rate of the change of pressure ratio with time. Furthermore, the effect of Mach number at the nozzle exit on hysteresis loop was investigated for two kinds of nozzle.     

A PCR assay for detection of acetic acid-tolerant lactic acid bacteria in acidic food products

A PCR assay for the detection of acetic acid-tolerant lactic acid bacteria in the genera of Lactobacillus and Pediococcus was developed in this study. Primers targeting the bacterial 16S rRNA gene were newly designed and used in this PCR assay. To determine the specificity of the assay, 56 different bacterial strains (of 33 genera), 2 fungi, 3 animals, and 4 plants were tested. Results were positive for most tested bacterial members of 16S rRNA gene-based phylogenetic groups (classified in the Lactobacillus casei and Pediococcus group), including Lactobacillus fructivorans, Lactobacillus brevis, Lactobacillus buchneri, Lactobacillus plantarum, and Lactobacillus paracasei. For all other bacterial strains and eukaryote tested, results were negative. Bacterial DNA for PCR was prepared with a simple procedure with the use of Chelex 100 resin from culture after growth in deMan Rogosa Sharpe broth (pH 6.0). To test this PCR assay for the monitoring of the acetic acid-tolerant lactic acid bacteria, L. fructivorans was inoculated into several acidic food as an indicator. Before the PCR, the inoculation of 10 to 50 CFU of bacteria per g of food was followed by a 28-h enrichment culture step, and the PCR assay allowed the detection of bacterial cells. Including the enrichment culture step, the entire PCR detection process can be completed within 30 h.     

Defective expression of the DNA mismatch repair protein, MLH1, alters G2-M cell cycle checkpoint arrest following ionizing radiation

A role for the Mut L homologue-1 (MLH1) protein, a necessary component of DNA mismatch repair (MMR), in G2-M cell cycle checkpoint arrest after 6-thioguanine (6-TG) exposure was suggested previously. A potential role for MLH1 in G1 arrest and/or G1-S transition after damage was, however, not discounted. We report that MLH1-deficient human colon carcinoma (HCT116) cells showed decreased survival and a concomitant deficiency in G2-M cell cycle checkpoint arrest after ionizing radiation (IR) compared with genetically matched, MMR-corrected human colon carcinoma (HCT116 3-6) cells. Similar responses were noted between murine MLH1 knockout compared to wild-type primary embryonic fibroblasts. MMR-deficient HCT116 cells or embryonic fibroblasts from MLH1 knockout mice also demonstrated classic DNA damage tolerance responses after 6-TG exposure. Interestingly, an enhanced p53 protein induction response was observed in HCT116 3-6 (MLH1+) compared with HCT116 (MLH1-) cells after IR or 6-TG. Retroviral vector-mediated expression of the E6 protein did not, however, affect the enhanced G2-M cell cycle arrest observed in HCT116 3-6 compared with MLH1-deficient HCT116 cells. A role for MLH1 in G2-M cell cycle checkpoint control, without alteration in G1, after IR was also suggested by similar S-phase progression between irradiated MLH1-deficient and MLH1-proficient human or murine cells. Introduction of a nocodazole-induced G2-M block, which corrected the MLH1-mediated G2-M arrest deficiency in HCT116 cells, clearly demonstrated that HCT116 and HCT116 3-6 cells did not differ in G1 arrest or G1-S cell cycle transition after IR. Thus, our data indicate that MLH1 does not play a major role in G1 cell cycle transition or arrest. We also show that human MLH1 and MSH2 steady-state protein levels did not vary with damage or cell cycle changes caused by IR or 6-TG. MLH1-mediated G2-M cell cycle delay (caused by either MMR proofreading of DNA lesions or by a direct function of the MLH1 protein in cell cycle arrest) may be important for DNA damage detection and repair prior to chromosome segregation to eliminate carcinogenic lesions (possibly brought on by misrepair) in daughter cells.     

Suppressing the effect of lightning-induced overvoltages by the combined use of surge arresters and overhead ground wires

Lightning voltage induced by nearby strokes is one of the causes of major overvoltages which threaten the insulation of power distribution lines. Surge arresters as well as an overhead ground wire usually are employed for the protection of equipment and line insulation on overhead power distribution lines. The fundamental mechanism of suppressing effects of overvoltages induced by nearby strokes has been demonstrated independently by one of the present authors for surge arresters and for an overhead ground wire. In this paper, the protective effect of the combination of surge arresters and overhead ground wire is analyzed. As a result, it is found that the effect of surge arresters used together with an overhead ground wire is almost the same as that of surge arresters alone.     

Decomposition of Benzene Using Ag/TiO2 Packed Plasma-Driven Catalyst Reactor: Influence of Electrode Configuration and Ag-Loading Amount

This paper describes the effects of electrode configuration and the loading amount of Ag catalyst on the decomposition of gas-phase benzene using plasma-driven catalyst (PDC) reactors. Modification of ground electrode brought out a great enhancement in the energy efficiency for benzene decomposition by reducing abnormal discharges outside the reactor tube. The data of carbon balance and the selectivity of CO₂ indicated that the Ag catalyst played an important role in the decomposition of benzene, especially for the intermediates. The larger the Ag-loading amounts on the TiO₂, the better the performance of benzene decomposition in terms of the carbon balance and the selectivity of CO₂. Formation of NO₂ and N₂O indicated that the maximum specific input energy applicable to the PDC reactor should be determined not only by the decomposition efficiency but also by the formation of nitrogen oxides.     

Creation of functional materials by use of a slow crystallization process in the nonequilibrium state. I. porous polyamide microparticles

Recently a number of investigations have focused on the self‐organization process from the nonequilibrium state as a new technique that may be used to develop many functional materials. In many cases, amorphous polymers were used and semicrystalline polymers were seldom used in spite of their importance. In this study, we basically investigated the crystal structure, crystalline process, and inner structures of polyamide by using slow phase separation and crystallization process from the nonequilibrium state of the polymer solution. We were able to observe the crystalline lamella growing twisted from the center of the particle. Between these lamella layers, narrow pores were created. From this investigation, we developed a new method to create functional materials of polyamide, the semicrystalline polymer. Porous spherical particles may be properly applied to functional materials such as adsorption materials, catalyst support materials, and so on. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 2428–2432, 2003     

Effects of highly hydrogenated soybean oil and cholesterol on plasma,liver cholesterol,and fecal steroids in rats

We investigated the relationship between dietary highly hydrogenated soybean oil (HSO) and cholesterol transport in rats. In the first study, to examine the effects of cholesterol transport of different concentrations of HSO in dietary oil, rats were given one of the three diets containing 0, 25, or 50% HSO in dietary oil with cholesterol (5 g/kg diet) or a diet without HSO and cholesterol for 22 d. Feeding the high concentration of HSO prevented the increase in plasma total cholesterol, hepatic total lipids, and cholesterol and the decrease in high-density lipoprotein-cholesterol, which were caused by dietary cholesterol. Moreover, HSO increased the fecal excretion, fecal lipids, and steroids in a dose-dependent manner. In the second study, to examine the effects on cholesterol transport of redistribution of stearic acid in the triacylglycerol species contained in HSO, rats were given one of the six diets containing HSO (distearoylmonoacylglycerol and tristearoylglycerol)-rich, monostearoylglycerol-rich, or palmitic acid-rich oil with/without cholesterol (5 g/kg diet), for 30 d. Whereas the accumulation of cholesterol in the body was reduced, cholesterol excretion was enhanced effectively in rats given the HSO-rich diet compared with rats given the monostearoylglycerol-rich diet. These results suggested that not only the high concentration of stearic acid but also its uneven distribution in HSO-triacylglycerol contributed to the reduction in intestinal cholesterol absorption in rats.