Phenotypic abnormalities observed in aged cloned mice from embryonic stem cells after long-term maintenance

Somatic/embryonic stem cell cloning has made it possible to produce an individual genomically identical to another individual. However, the cloned animals have a variety of abnormalities caused by the aberrant gene modification, with insufficient reprogramming in cloning. We previously reported abnormalities in cloned mice at birth. In this study, we examined what abnormalities could be seen in cloned mice after long-term maintenance. The aged cloned mice showed multiple abnormalities: increase of body weight, some phenotypic abnormalities in the kidneys, testes and thymus, and lower urea nitrogen in their serum biochemical values. The kidneys of all cloned mice were hypertrophied, with a metamorphic or whitish appearance. The multiple lesions, including the enlarged renal pelvis and distension of the renal veins in histology, might be the result of urine accumulation by urinary tract obstruction. The testes of the cloned mice were atrophied, and showed no sperm formation in histology. In contrast, the thymus was rather hypertrophied, and a comparably increased number of lymphocytes were observed in the medulla, consisting mainly of T cells. By conducting a progeny test between the cloned mice, it was confirmed that these abnormalities in the aged cloned mice were not transmitted to their offspring, indicating that the incomplete reprogramming in clones might be in part responsible for the abnormalities detected in aged clones. These results indicate that the postnatal abnormalities observed in aged cloned mice are varied and can be restored through the germ line.     

Preparation of a single-chain variable fragment and a recombinant antigen-binding fragment against the anti-malarial drugs, artemisinin and artesunate, and their application in an ELISA

Two different recombinant antibodies, a single-chain variable fragment (scFv) and an antigen-binding fragment (Fab), were prepared against artemisinin (AM) and artesunate (AS) and were developed for use in an enzyme-linked immunosorbent assay (ELISA). The recombinant antibodies, which were derived from a single monoclonal antibody against AM and AS (mAb 1C1) prepared by us, were expressed by Escherichia coli cells and their reactivity and specificity were characterized. As a result, to obtain sufficient signal in indirect ELISA, a much greater amount of a first antibody was needed in the use of scFv due to the differences of the secondary antibody and conformational stability. Therefore, we focused on the development of the recombinant Fab antibodies and applied it to indirect competitive ELISA. The specificity of the Fab was similar to that of mAb 1C1 in that it showed specific reactivity toward AM and AS only. The sensitivity of the icELISA (0.16 μg/mL to 40 μg/mL for AM and 8.0 ng/mL to 60 ng/mL for AS) was sufficient for analysis of antimalarial drugs, and its utility for quality control of analysis of Artemisia spp. was validated. The Fab expression and refolding systems provided a good yield of high-quality antibodies. The recombinant antibody against AM and AS provides an essential component of an economically attractive immunoassay and will be useful in other immunochemical applications for the analysis and purification of antimalarial drugs.     

Physician-received scatter radiation with angiography systems used for interventional radiology: comparison among many X-ray systems

Radiation protection for interventional radiology (IR) physicians is very important. Current IR X-ray systems tend to use flat-panel detectors (FPDs) rather than image intensifiers (IIs). The purpose of this study is to test the hypothesis that there is no difference in physician-received scatter radiation (PRSR) between FPD systems and II systems. This study examined 20 X-ray systems in 15 cardiac catheterisation laboratories (11 used a FPD and 9 used an II). The PRSR with digital cineangiography and fluoroscopy were compared among the 20 X-ray systems using a phantom and a solid-state-detector electronic pocket dosemeter. The maximum PRSR exceeded the minimum PRSR by ~12-fold for cineangiography and ~9-fold for fluoroscopy. For both fluoroscopy and digital cineangiography, the PRSR had a statistically significant positive correlation with the entrance surface dose (fluoroscopy, r = 0.87; cineangiography, r = 0.86). There was no statistically significant difference between the average PRSR of FPDs and IIs during either digital cineangiography or fluoroscopy. There is a wide range of PRSR among the radiography systems evaluated. The PRSR correlated well with the entrance surface dose of the phantom in 20 X-ray units used for IR. Hence, decreasing the dose to the patient will also decrease the dose to staff.     

Preparation of core–shell particles by dispersion polymerization with a poly(ethylene oxide) macroazoinitiator

A macroazoinitiator (MAI) containing a poly(ethylene oxide) (PEO) block was used with a methyl methacrylate monomer to prepare polymer particles in ethanol/H₂O solutions. The effects of the monomer/MAI ratio (RMI) and H₂O content in the solutions on the molecular weight, particle diameters, and chemical structure of the resulting polymer particles were investigated. The reaction mixtures showed three kinds of states, which were milky colloid solutions, macrogels and/or precipitations, and clear solutions. The colloid solutions were obtained in the solutions with an H₂O content of about 50–90 vol % and a RMI of 20–400. In the colloid solutions, core–shell nanospheres consisting of PEO shells and poly(methyl methacrylate) (PMMA) cores were predominantly obtained. In the specific conditions close to the area of gel and/or precipitation formation, particles connected about 0.5–5 μm in length were obtained. Multiblock copolymers nanospheres tended to be obtained with lower RMIs, and PMMA‐PEO‐PMMA tri‐bloc and/or PMMA‐PEO di‐block copolymer nanospheres were obtained with higher RMIs. The solubility of the monomer and the generated polymer in solutions may have affected the polymerization development and the state of the products. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Isotopomer analysis of production and consumption mechanisms of N2O and CH4 in an advanced wastewater treatment system

Wastewater treatment processes are believed to be anthropogenic sources of nitrous oxide (N(2)O) and methane (CH(4)). However, few studies have examined the mechanisms and controlling factors in production of these greenhouse gases in complex bacterial systems. To elucidate production and consumption mechanisms of N(2)O and CH(4) in microbial consortia during wastewater treatment and to characterize human waste sources, we measured their concentrations and isotopomer ratios (elemental isotope ratios and site-specific N isotope ratios in asymmetric molecules of NNO) in water and gas samples collected by an advanced treatment system in Tokyo. Although the estimated emissions of N(2)O and CH(4) from the system were found to be lower than those from the typical treatment systems reported before, water in biological reaction tanks was supersaturated with both gases. The concentration of N(2)O, produced mainly by nitrifier-denitrification as indicated by isotopomer ratios, was highest in the oxic tank (ca. 4000% saturation). The dissolved CH(4) concentration was highest in in-flow water (ca. 3000% saturation). It decreased gradually during treatment. Its carbon isotope ratio indicated that the decrease resulted from bacterial CH(4) oxidation and that microbial CH(4) production can occur in anaerobic and settling tanks.     

A high-precision low-voltage bipolar current mirror circuit and its compensation for stability

A current mirror circuit is widely utilized as an important building block in analogue signal processing circuits. This paper describes a high-precision low voltage bipolar current mirror circuit. Well known current mirror circuits, such as a simple current mirror and a Wilson current mirror, have a trade-off between low-voltage operation and accuracy of the current gain. The accuracy of the proposed current mirror is high in spite of low current amplification factor (β) of transistors and large current output (or a large number of the multiple output). The current mirror circuit can operate at a 1V or less supply voltage. The high accuracy is realized by negative current feedback with high gain. Thus, the stability of the circuit and compensation methods are discussed. PSPICE simulation shows that the proposed current mirror circuit compensated using Miller components composed of a capacitor and a resistor is stable and can operate at a 1V supply voltage.     

Measurement of surface dryout near heating surface at high heat fluxes in subcooled pool boiling

The authors have conducted measurements of liquid–vapor behavior in the vicinity of a heating surface for saturated and subcooled pool boiling on an upward-facing copper surface by using a conductance probe method. A previous paper [A. Ono, H. Sakashita, Liquid–vapor structure near heating surface at high heat flux in subcooled pool boiling, Int. J. Heat Mass Transfer 50 (2007) 3481–3489] reported that thicknesses of a liquid rich layer (a so-called macrolayer) forming in subcooled boiling are comparable to or thicker than those formed near the critical heat flux (CHF) in saturated boiling. This paper examines the dryout behavior of the heating surface by utilizing the feature that a thin conductance probe placed very close to the heating surface can detect the formation and dryout of the macrolayer. It was found that the dryout of the macrolayer formed beneath a vapor mass occurs in the latter half of the hovering period of the vapor mass. Two-dimensional measurements conducted at 121 grid points in a 1-mm × 1-mm area at the center of the heating surface showed that the dryout commences at specific areas and spreads over the heating surface as the heat flux approaches the CHF. Furthermore, transient measurements of wall void fractions from nucleate boiling to transition boiling were conducted under the transient heating mode, showing that the wall void fraction has small values (<10%) in the nucleate boiling region, and then steeply increases in the transition boiling region. These findings strongly suggest that the macrolayer dryout model is the most appropriate model of the CHF for saturated and subcooled pool boiling of water on upward facing copper surfaces.     

Temperature measurements near a heating surface at high heat fluxes in subcooled pool boiling

In previous papers (Int J Heat Mass Transfer, 2008;50:3481–3489, 2009;52: 814–821), the authors conducted measurements of liquid–vapor structures in the vicinity of a heating surface for subcooled pool boiling on an upward‐facing copper surface by using a conducting probe method. We reported that the macrolayer dryout model is the most appropriate model of the CHF and that the reason why the CHF increases with increasing subcooling is most likely that a thick macrolayer is able to form beneath large vapor masses and the lowest heat flux of the vapor mass region shifts towards the higher heat flux. To develop a mechanistic model of the CHF for subcooled boiling, therefore, it is necessary to elucidate the effects of local subcooling on boiling behaviors in the vicinity of a heating surface. This paper measured local temperatures close to a heating surface using a micro‐thermocouple at high heat fluxes for water boiling on an upward‐facing surface in the 0 to 40 K range of subcooling. A value for the effective subcooling, defined as the local subcooling during the period while vapor masses are being formed was estimated from the detected bottom peaks of the temperature fluctuations. It was established that the effective subcooling adjacent to the surface remains at considerably lower values than the bulk liquid subcooling. This suggests that, from nucleation to coalescence, the subcooling of a bulk liquid has a smaller effect on the behavior of primary bubbles than the extent of the subcooling would appear to suggest. An empirical correlation of the effective subcooling is proposed to provide a step towards quantitative modeling of the CHF for subcooled boiling. © 2009 Wiley Periodicals, Inc. Heat Trans Asian Res; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/htj.20277