Aging and thermal stability of Mg/SiC and Mg/Y2O3 reflection multilayers in the 25-35 nm region

Reflection measurements in the 25-35 nm region were made for Mg/SiC and Mg/Y2O3 multilayers kept in a low-humidity atmosphere for 4 or 5 years. Aged Mg/SiC multilayers keep their reflectances, and the reflectance value at 31.2 nm is 0.44 at 10 degrees of the normal angle of incidence. Aged Mg/Y2O3 multilayers change reflectance as top layer materials, and the best value at 30.1 nm is 0.40 at 10 degrees. Reflection measurements are also made for Mg-based multilayers that are annealed from room temperature to 400 degrees C at 50 degrees C intervals. Both multilayers keep their reflectance at annealing temperatures of 200 degrees C. These results suggest that both Mg-based multilayers can be applied to practical optics.     

Prediction of cutting forces and machining error in ball end milling of curved surfaces -II experimental verification

This paper presents the results of a series of experiments performed to examine the validity of a theoretical model for evaluation of cutting forces and machining error in ball end milling of curved surfaces. The experiments are carried out at various cutting conditions, for both contouring and ramping of convex and concave surfaces. A high precision machining center is used in the cutting tests. In contouring, the machining error is measured with an electric micrometer, while in ramping it is measured on a 3-coordinate measuring machine. The results show that in contouring, the cutting force component that influences the machining error decreases with an increase in milling position angle, while in ramping, the two force components that influence the machining error are hardly affected by the milling position angle. Moreover, in contouring, high machining accuracy is achieved in “Up cross-feed, Up cut” and “Down cross-feed, Down cut” modes, while in ramping, high machining accuracy is achieved in “Left cross-feed, Downward cut” and “Right cross-feed, Upward cut” modes. The theoretical and experimental results show reasonably good agreement.     

DNA taken into Bacillus subtilis competent cells by lysed-protoplast transformation is not ssDNA but dsDNA

Competent Bacillus subtilis incorporates whole-genome DNA (4215 kb) from the protoplast lysate of B. subtilis subtilis [Akamatsu, T. and Taguchi, H., Biosci. Biotechnol. Biochem., 65, 823-829 (2001)]. A continuous incorporated DNA is longer than 1500 kb [J. Biosci. Bioeng., 101, 257-262 (2006)]. Whether the incorporated DNA is single-stranded (ssDNA) or double-stranded DNA (dsDNA) has been studied by examining the transforming activity of the incorporated DNA. B. subtilis BEST7027 was used as the donor strain, which has a heterologous region consisting of the 145 kb region of the Synechocystis sp. PCC6803 genome and erm gene. The donor DNA was transferred to a wild-type or a recA recipient strain (AYG2 or SYN9), and protoplast lysate was prepared from the transformants and used as the donor DNA source for the second recipient strain (AU1 or AV1). The intergenote region showed a significant transforming activity. When DNase I was added to both cells collected from the first transformation mixture and the following protoplastization, the result was similar to that obtained without DNase I. All of the observations strongly suggest that the incorporated DNA is dsDNA, and the transformation of competent B. subtilis by DNA in protoplast lysate is different from that by purified DNA taken up conventionally.     

Fate of transforming bacterial genome following incorporation into competent cells of Bacillus subtilis: a continuous length of incorporated DNA

In contrast to the conventional transformation of Bacillus subtilis using purified DNA, those using DNA in lysed protoplasts have a high transformation efficiency and enable whole-genome transfer into competent B. subtilis [Akamatsu, T. and Taguchi, H., Biosci. Biotechnol. Biochem., 65, 823-829 (2001)]. Here, we examined the length of incorporated continuous DNA by analyzing the cotransfer ratio with selected and unselected markers, on the basis of a new experimental design. The cotransfer ratio of a selected marker with an unselected marker on the opposite side of the genetic map of the B. subtilis chromosome was about 5.6% and could be interpreted as congression (double transformation) ratio. In the wild-type strain, the cotransfer ratio of cysA (113 kb position on 4215 kb of B. subtilis chromosome) with metC (1384 kb) and leuB (2891 kb) was 0.77%, twice the value (5.6% x 5.6%=0.31%) calculated from the congression ratio. Moreover, in a genetic background, the cotransfer ratios of metC with cysA and leuB, and metC with cysA and arg1 (3012 kb) were 2.7% and 7.2%, respectively. These results strongly suggest that the length of continuous DNA incorporated into B. subtilis is most probably greater than 1271 kb. When the DNA from the protoplast lysate was fragmented by mixing, the cotransfer ratios of arg1 with metC, and arg1 with metC and trpC (2374 kb) were 2.8% and 0.16%, respectively. A high cotransfer ratio (2.7-7.2%) could not, therefore, be obtained using the fragmented DNA. Based on these observations, we propose a working hypothesis on the mechanism of the transformation of competent B. subtilis by DNA in protoplast lysates (LP transformation).     

Dendrimer-based nanoprobe for dual modality magnetic resonance and fluorescence imaging

A novel PAMAM dendrimer-based nanoprobe for dual magnetic resonance and fluorescence imaging modalities was synthesized. Fluorescence studies revealed that Gd(III) complexation to the probe has no effect on the quantum yield; however, increases in the dye content resulted in partial quenching. The potential of the new nanoprobe, G6-(Cy5.5)(1.25)(1B4M-Gd)(145), as a dual modality imaging agent was demonstrated in vivo by the efficient visualization of sentinel lymph nodes in mice by both MRI and fluorescence imaging modalities.     

Role of ComEA in DNA uptake during transformation of competent Bacillus subtilis

The role of the competence protein ComEA in DNA uptake during transformation of competent Bacillus subtilis was analyzed by lysed-protoplast transformation (LP transformation). A comEA deletion mutant was constructed by a fusion polymerase chain reaction. Transformants of the mutant were obtained by LP transformation at a frequency of 1.1 × 10(2) transformants per μg DNA, representing a low relative efficiency of transformation [RET (mutant/wild type)] of 2.7 × 10(-6). This implied an important role of the protein during DNA uptake. When analyzing LP transformation of comEA with a plasmid (5.7 kb), a similar RET (mutant/wild type) of 5.6 × 10(-5) was obtained. Following addition of DNA into the comEA mutant culture, the number of transformants increased at a rate of 0.5 transformants/min, which was very low compared with the wild-type (6.9×10(4) transformants/min). However, even in the comEA mutant, DNA uptake began immediately after addition of DNA. Using co-transformation analysis of the comEA mutant, short linkages at distances of 2-156 kb could be detected, but not long linkages at distances of 671-1662 kb. Taken together, the results indicate that ComEA plays an important role in the transfer of transforming DNA into the DNA channel and in controlling the rate of DNA uptake.     

C57BL/KsJ-db/db-Apc Mice Exhibit an Increased Incidence of Intestinal Neoplasms

The numbers of obese people and diabetic patients are ever increasing. Obesity and diabetes are high-risk conditions for chronic diseases, including certain types of cancer, such as colorectal cancer (CRC). The aim of this study was to develop a novel animal model in order to clarify the pathobiology of CRC development in obese and diabetic patients. We developed an animal model of obesity and colorectal cancer by breeding the C57BL/KsJ-db/db (db/db) mouse, an animal model of obesity and type II diabetes, and the C57BL/6J-Apc(Min/+) (Min/+) mouse, a model of familial adenomatous polyposis. At 15 weeks of age, the N9 backcross generation of C57BL/KsJ-db/db-Apc(Min/+) (db/db-Min/+) mice developed an increased incidence and multiplicity of adenomas in the intestinal tract when compared to the db/m-Min/+ and m/m-Min/+ mice. Blood biochemical profile showed significant increases in insulin (8.3-fold to 11.7-fold), cholesterol (1.2-fold to 1.7-fold), and triglyceride (1.2-fold to 1.3-fold) in the db/db-Min/+ mice, when compared to those of the db/m-Min/+ and m/m-Min/+ mice. Increases (1.4-fold to 2.6-fold) in RNA levels of insulin-like growth factor (IGF)-1, IRF-1R, and IGF-2 were also observed in the db/db- Min/+ mice. These results suggested that the IGFs, as well as hyperlipidemia and hyperinsulinemia, promoted adenoma formation in the db/db-Min/+ mice. Our results thus suggested that the db/db-Min/+ mice should be invaluable for studies on the pathogenesis of CRC in obese and diabetes patients and the therapy and prevention of CRC in these patients.     

Inhibition effect of CO on hydrogen permeability of Pd–Ag membrane applied in a microchannel module configuration

The surface adsorption effect of CO on the hydrogen permeability of a 12.5 micron-thick Pd₇₇Ag₂₃ membrane has been evaluated quantitatively under experimental conditions close to the operating conditions of the highly-efficient membrane reformer (MRF) system developed by Tokyo Gas. The permeability of the membrane was measured in the conditions of CO concentration between 1 and 5 vol.% at a temperature and pressure of up to 500 °C and 0.6 MPa, respectively. High feed flow rates and a microchannel module configuration were applied in the flux measurements to ensure that the results are obtained with limited influence of concentration polarization adjacent to the membrane surface and hydrogen depletion along the microchannel length. While the CO inhibition effect was close to negligible at 500 °C, it was significant at lower temperatures. At a feed pressure of 0.2 MPa, the CO inhibition effect was only 0.2% at a CO concentration of 1 vol.% and the effect was 3.6% at a CO concentration of 5 vol.% at 500 °C. The CO inhibition effect were 3.4% for 1 vol.% CO and 14.1% for 5 vol.% CO at 400 °C. Measurements were also carried out at a high feed pressure of 0.6 MPa to evaluate the pressure dependence of the CO inhibition effect. The CO inhibition effect decreased to 0.7% at a CO feed concentration of 5 vol.% at 500 °C. Lower CO inhibition effect were also observed at 450 and 400 °C compared to the data obtained with the feed pressure of 0.2 MPa, while the inhibition levels were almost the same at 350 °C. Though the CO inhibition effect is larger at a lower feed pressure of 0.2 MPa, the effect was only 0.2% at 1 vol.% CO at 500 °C, which is close to the operating conditions of the MRF system. This study quantitatively revealed that the CO inhibition effect on hydrogen flux is extremely small when the membrane is operated at temperatures equal to or higher than 500 °C, even for state-of-the-art thin membranes. The performance of the Tokyo Gas MRF seems thus mainly limited by concentration polarization effects.     

Enhanced Development of Azoxymethane-Induced Colonic Preneoplastic Lesions in Hypertensive Rats

Metabolic syndrome is associated with an increased risk of colorectal cancer. This study investigated the impact of hypertension, a component of metabolic syndrome, on azoxymethane (AOM)-induced colorectal carcinogenesis using SHRSP/Izm (SHRSP) non-diabetic/hypertensive rats and SHRSP.Z-Lepr^fa/IzmDmcr (SHRSP-ZF) diabetic/hypertensive rats. Male 6-week-old SHRSP, SHRSP-ZF, and control non-diabetic/normotensive Wister Kyoto/Izm (WKY) rats were given 2 weekly intraperitoneal injections of AOM (20 mg/kg body weight). Two weeks after the last injection of AOM, the SHRSP and SHRSP-ZF rats became hypertensive compared to the control WKY rats. Serum levels of angiotensin-II, the active product of the renin-angiotensin system, were elevated in both SHRSP and SHRSP-ZF rats, but only the SHRSP-ZF rats developed insulin resistance, dyslipidemia, and hyperleptinemia and exhibited an increase in adipose tissue. The development of AOM-induced colonic preneoplastic lesions and aberrant crypts foci, was significantly accelerated in both SHRSP and SHRSP-ZF hypertensive rats, compared to WKY normotensive rats. Furthermore, induction of oxidative stress and exacerbation of inflammation were observed in the colonic mucosa and systemically in SHRSP and SHRSP-ZF rats. Our findings suggest that hypertension plays a role in the early stage of colorectal carcinogenesis by inducing oxidative stress and chronic inflammation, which might be associated with activation of the renin-angiotensin system.