In vitro construction and screening of a Burkholderia cepacia lipase library using single-molecule PCR and cell-free protein synthesis

A mutant library of Burkholderia cepacia lipase KWI-56 was constructed on microplates by a cell-free process and tested with a chromogenic assay. This high-throughput construction system can be used to screen mutant proteins based on their catalytic activity.     

Interactive segmentation of the cerebral lobes with fuzzy inference in 3T MR images.

Measurement of volume and surface area of the frontal, parietal, temporal and occipital lobes from magnetic resonance (MR) images shows promise as a method for use in diagnosis of dementia. This article presents a novel computer-aided system for automatically segmenting the cerebral lobes from 3T human brain MR images. Until now, the anatomical definition of cerebral lobes on the cerebral cortex is somewhat vague for use in automatic delineation of boundary lines, and there is no definition of cerebral lobes in the interior of the cerebrum. Therefore, we have developed a new method for defining cerebral lobes on the cerebral cortex and in the interior of the cerebrum. The proposed method determines the boundaries between the lobes by deforming initial surfaces. The initial surfaces are automatically determined based on user-given landmarks. They are smoothed and deformed so that the deforming boundaries run along the hourglass portion of the three-dimensional shape of the cerebrum with fuzzy rule-based active contour and surface models. The cerebrum is divided into the cerebral lobes according to the boundaries determined using this method. The reproducibility of our system with a given subject was assessed by examining the variability of volume and surface area in three healthy subjects, with measurements performed by three beginners and one expert user. The experimental results show that our system segments the cerebral lobes with high reproducibility.     

Space charge behavior in low density polyethylene at pre-breakdown

It has been known that the electrical breakdown of insulating materials is strongly affected by the formation of space charge in the bulk of the materials. Many researchers have attempted to study the relationship between the space charge and the breakdown; however, it has not been clarified yet. Although the pulsed electroacoustic (PEA) method has been widely used to observe space charge profiles, previous works have not shown clear evidence of the influence of the space charge on the breakdown. Therefore, we have developed a new PEA system with an interval of 0.5 ms to observe the space charge distribution continuously under the ultra-high electric field. Using this system, we observed the space charge dynamics in the low-density polyethylene (LDPE) at and around the breakdown. We also investigated the dependence of the applied electric field on space charge behavior. From the results, it is found that the injected charge packet moved faster and deeper under a relatively lower electric field rather than that under a higher electric field. Furthermore, we found that the maximum electric field in each specimen was almost the same when the breakdown happened in a specimen.     

Statistical analysis of correlated phase error in transmission characteristics of arrayed-waveguide gratings

Transmission characteristics of arrayed waveguide gratings (AWGs) are significantly influenced by phase errors in a waveguide array, which are easily caused by the variation of fabrication process. In this paper, the statistical analysis of transmission characteristics is presented using correlation and spectral density of phase errors. A simple statistical approach, based on a simplified model of a waveguide array, is newly introduced to clarify statistical behavior of phase errors in AWGs. Spectral density is modeled from measured phase errors, and then average transmission characteristics are simulated for AWGs with different channel spacing. Average transmission characteristics, simulated with the spectral density model, almost describe measured results in the cases of various channel spacing. It is also indicated that an AWG may have larger adjacent-channel crosstalk with narrower channel spacing because the adjacent-channel crosstalk is affected by phase errors with a smaller spatial frequency.     

Thermodynamics of iron oxide in Fe x O-dilute CaO + Al2O3 + MgO + Fe x O slags at 1873 K

In order to determine the ferrous and ferric ion capacities: 3

Molecular analysis of mutations induced by acrolein in human fibroblast cells using supF shuttle vector plasmids

Types of mutations induced by acrolein in the supF gene on the shuttle vector plasmid pMY189 replicated in normal human fibroblast cells were examined. Base sequence analysis of 92 plasmids with mutations in the supF gene revealed that the majority of the mutations were base substitutions (76%) and the others were deletions and insertions (24%). Single base substitutions were most frequently found (46%), while multiple base substitutions were 18% and tandem (two adjacent) base substitutions were 12% of the mutations. Of the base substitution mutations, G:C to T:A transversions were 44% and G:C to A:T transitions were 24%. The mutations were distributed not randomly but located at several hotspots. Acrolein produced DNA intra-strand cross-links between guanine residues, which might be responsible for rather high induction of the tandem base substitution mutations.     

MaMi, a human endometrial stromal sarcoma cell line that constitutively produces interleukin-6, interleukin-8, and monocyte chemoattractant protein-1

BACKGROUND: Uterine endometrial stromal sarcoma is a rare neoplasm with a morphology that closely resembles that of the proliferative endometrial stroma. To understand its pathologic characteristics, we established a novel cell line, MaMi, from a primary culture of an endometrial stromal sarcoma obtained from a 65-year-old Japanese woman. METHODS: We observed the morphology of MaMi cells and performed immunohistochemical analysis on the primary tumor and transplants in nude mice. Prolactin, insulin-like growth factor-binding protein-1, interleukin (IL)-6, IL-8, monocyte chemoattractant protein-1, intercellular adhesion molecule-1, E-selectin, vascular cell adhesion molecule-1, and fibronectin production in the culture medium of MaMi cells were also examined. RESULTS: MaMi cells were shown to exhibit a fibroblast-like morphology in vitro, and they adopted a more elongated appearance in response to 12-O-tetradecanoyl phorbol-13-acetate (TPA). On injection into nude mice, the cells gave rise to subcutaneous tumors. Immunohistologically, both the primary tumor and MaMi cell-induced tumors stained positively with antibodies to neuron-specific enolase or vimentin. MaMi cells constitutively produced IL-6, IL-8, and monocyte chemoattractant protein-1 in vitro. Interleukin-1beta, (100 pmol/L), tumor necrosis factor-alpha (1 nmol/L), and lipopolysaccharide (1 microg/mL) each increased the release of IL-6, IL-8, and monocyte chemoattractant protein-1 by MaMi cells. TPA (10 nmol/L) also stimulated the production of IL-6 and IL-8 by these cells, but inhibited that of monocyte chemoattractant protein-1. CONCLUSIONS: We demonstrated that MaMi cells closely resemble proliferative endometrial stromal cells not only morphologically, but also functionally. This cell line may prove valuable in understanding the role of cytokines produced by tumor cells in the pathogenesis of endometrial stromal sarcoma and may also be useful as an in vitro model of functioning endometrial stromal cells.     

Improvements in finite-element simulation for stamping and application to the forming of laser-welded blanks

During stamping-die design, the formability in sheet-metal forming process has been evaluated by the geometrical functions in ‘Die-Face CAD’, which has been developed and improved by Toyota Motor Corporation. When evaluation by these functions is difficult, formability has been estimated by performing experiments using test dies in which the forming defects are similar to those in the actual process.

A numerical method has been developed in order to substitute numerical analysis for experiments using test dies for the accurate prediction of defects in sheet-metal forming. The elastic-plastic FEM with the commercial code ‘JNIKE3D’ has been improved in the areas of: (1) the material constitutive equation; (2) the consideration of the pressure distribution on the blank-holder; and (3) the evaluation of breakage initiation. Using the improved method, the square-cup drawing process and the hemming process have been analyzed. Numerical results for strain, breakage initiation, and hemming deflection were in good agreement with experimental results. The formability of laser-welded blanks and the most efficient process to form them were evaluated also using the improved method.     

Specific tandem GG to TT base substitutions induced by acetaldehyde are due to intra-strand crosslinks between adjacent guanine bases

Acetaldehyde is present in tobacco smoke and automotive exhaust gases, is produced by the oxidation of ethanol, and causes respiratory organ cancers in animals. We show both the types and spectra of acetaldehyde-induced mutations in supF genes in double- and single-stranded shuttle vector plasmids replicated in human cells. Of the 101 mutants obtained from the double-stranded plasmids, 63% had tandem base substitutions, of which the predominant type is GG to TT transversions. Of the 44 mutants obtained from the single-stranded plasmids, 39% had tandem mutations that are of a different type than the double-stranded ones. The GG to TT tandem substitutions could arise from intra-strand crosslinks. Our data indicate that acetaldehyde forms intra- as well as inter-strand crosslinks between adjacent two-guanine bases. Based upon the following observations: XP-A protein binds to acetaldehyde-treated DNA, DNA excision repair-deficient xeroderma pigmentosum (XP) cells were more sensitive to acetaldehyde than the repair-proficient normal cells, and a higher frequency of acetaldehyde-induced mutations of the shuttle vectors was found in XP cells than in normal cells, we propose that the DNA damage caused by acetaldehyde is removed by the nucleotide excision repair pathway. Since treatment with acetaldehyde yields very specific GG to TT tandem base substitutions in DNA, such changes can be used as a probe to identify acetaldehyde as the causal agent in human tumors.