Development of a homogeneous competitive immunoassay for phosphorylated protein antigen based on the enhanced fluorescence resonance energy transfer technology

We describe a homogeneous competitive immunoassay for a phosphorylated protein antigen. The assay takes advantage of the enhanced fluorescence resonance energy transfer (FRET) technology, which has a unique characteristic that the FRET signal is increased by the specific interaction of two fluorolabeled leucine zippers. We chose extracellular signal-regulated kinase (ERK) as a model antigen and constructed two molecular probes in which either anti-phosphorylation site antibody or the antigen peptide was chemically conjugated to the enhanced FRET probes. While these molecular probes indicated sufficient FRET signal without antigen, they displayed a significant change in the fluorescent spectrum by mixing with phosphorylated antigens. With this competitive enhanced FRET immunoassay, a phosphorylated ERK concentration within the range from 15 nM to 250 nM could be determined. Because the assay is very simple, it would be applied to not only in vitro assay but also in vivo detection of protein phosphorylation.     

Validation of MAAP model enhancement for Fukushima Dai-ichi accident analysis with Phenomena Identification and Ranking Table (PIRT)

The Modular Accident Analysis Program (MAAP) model enhancement items to improve the simulation capability for molten corium behavior in the accidents at the Fukushima Dai-ichi nuclear power plants were validated with the Phenomena Identification and Ranking Table (PIRT). The importance ranks of the identified phenomena were evaluated for each time phase through brainstorming and discussion with the experts in the Atomic Energy Society of Japan and the members of the MAAP model enhancement project. When the current MAAP evaluation models were reviewed with the PIRT, it is found that 95 of the 386 high-ranked phenomena were not considered in MAAP 5.0.1. While 62 of these phenomena will have been addressed in the MAAP enhancement project and 25 others are not suitable to be analyzed by MAAP, 8 important phenomena should be considered in post-MAAP enhancement project with additional experiments or fundamental studies.     

Novel Enantiocomplementary C2‐Symmetric Chiral Bis(imidazoline) Ligands: Highly Enantioselective Friedel–Crafts Alkylation of Indoles with Ethyl 3,3,3‐Trifluoropyruvate

Enantioselective Friedel–Crafts alkylations of a variety of indoles with ethyl 3,3,3‐trifluoropyruvate catalyzed by novel chiral m‐phenylenebis(imidazoline)‐copper(II) complexes or the bis(imidazoline)‐achiral acid combination afforded products with high enantioselectivity. Both enantiomers of indole derivatives can be prepared with high enantioselectivities by tuning the N‐substituents of the imidazoline.     

Functional analyses of neutrophil-like differentiated cell lines under a hyperglycemic condition

Diabetic patients are prone to severe bacterial infections. The functional alterations of neutrophils by hyperglycemia are thought to be partially responsible for such infections. In this study, we investigated the functional changes of neutrophil-like differentiated cell lines (dHL-60, dTHP-1, and dNB-4) by treatment with 5.5 mM, 11 mM, or 35 mM of glucose. In dHL-60 cells, the incubation with high glucose (35 mM) resulted in the enhancement of cell aggregation, the suppression of cellular fragility, the induction of reactive-oxygen species (ROS) production by phorbol myristate acetate (PMA) stimulation, and the impairment of phagocytosis. In dTHP-1 cells, the treatment with higher glucose generated the suppression of cellular fragility and extremely impaired phagocytosis (by 35 mM), and induced ROS production due to PMA stimulation (by 11 mM). Furthermore, the higher glucose exposure to dNB-4 cells enlarged intracellular vacuoles (by 35 mM) and induced ROS production due to PMA stimulation (by 11 mM). Since the ROS generation of those cells was enhanced only after PMA stimulation under the higher glucose conditions, glucose may have a priming effect rather than a triggering effect. These extraordinary sensitivities caused by the higher glucose treatments may reflect the dysfunction or overactivation of neutrophils.     

Smear growth on head slider surface from siloxane outgas on heat assisted magnetic recording

Microsystem Technologies - In this study, the smear by the lubricant and siloxane pickup at the laser heating in the heat assisted magnetic recording (HAMR) was evaluated using our developed...     

Image content adaptive color breakup index for field sequential color displays using a dominant visual saliency method

An index that can predict the perceptual visibility of color breakup for varying image content is valuable in field sequential color displays, whereas the current indices are usually for fixed patterns. To solve this problem, an image database containing 25 diverse reference images and 125 test cases with various color breakup visibility was first established. Next, visual experiments using a 240‐Hz liquid crystal display were performed to acquire the subjective color breakup scores of the test cases. A theorem based on visual saliency theory was proposed that the color breakup perception is mainly determined by the image regions with visual saliency values higher than a certain threshold, called the dominant visual saliency regions. A computational model based on this theorem was developed to obtain objective color breakup scores of the test cases from retinal images with and without color breakup. An analysis of the objective and subjective results revealed a Pearson linear correlation coefficient as high as 0.82, which matches the top‐level image quality assessment algorithms. Finally, the proposed color breakup index was used to benchmark against several mainstream field sequential color algorithms to determine their performances in color breakup suppression.     

Grafting of branched polymers onto nano-sized silica surface: Postgrafting of polymers with pendant isocyanate groups of polymer chain grafted onto nano-sized silica surface

To control the surface wettability of nano-sized silica surface, the postgrafting of hydrophilic and hydrophobic polymers to grafted polymer chains on the surface was investigated. Polymers having blocked isocyanate groups were successfully grafted onto nano-sized silica surface by the graft copolymerization of methyl methacrylate (MMA) with 2-(O-[1′-methylpropylideneamino]caboxyamino)ethyl methacrylate (MOIB) initiated by azo groups previously introduced onto the surface. The blocked isocyanate groups of poly(MMA-co-MOIB)-grafted silica were stable in a desiccator, but isocyanate groups were readily regenerated by heating at 150 °C. The hydrophilic polymers, such as poly(ethylene glycol) (PEG) and poly(ethyleneimine) (PEI), were postgrafted onto the poly(MMA-co-MOIB)-grafted silica by the reaction of functional groups of PEG and PEI with pendant isocyanate groups of poly(MMA-co-MOI)-grafted silica to give branched polymer-grafted silica. The percentage of grafting increased with increasing molecular weight of PEG, but the number of postgrafted chain decreased, because of steric hindrance. The hydrophobic polymers, such as poly(dimethylsiloxane) were also postgrafted onto poly(MMA-co-MOI)-grafted silica. It was found that the grafting of hydrophobic polymer and the postgrafting of hydrophilic polymer branches readily controls the wettability of silica surface to water.     

The structure of the core polyol of the ether lipids fromSulfolobus acidocaldarius

The major ether-type lipid structures ofSulfolobus acidocaldarius (ATCC33909) were composed of caldarchaeol and calditoglycerocaldarchaeol. However, the characterization by nuclear magnetic resonance spectroscopy and mass spectrometry showed that the structure of calditol in calditoglycerocaldarchaeol is not nonitol, 2-(1′,2′,3′-trihydroxypropyl)1,2,3,4,5,6-hexahydroxyhexane, but 2-hydroxymethyl-1-(2,3-dihydroxypropoxy),2,3,4,5-cyclopentanetetraol with an ether linkage in the molecule. Such an intermolecular ether linkage was resistant, to BCl₃ treatment, but nonresistant to 57% HI degradation treatment conducted at 100°C for 60 h, producting 2-hydroxymethyl-1,2,3,4,5-cyclopentanepentaol from calditol as reaction product. Further, it was confirmed that the structure of calditol is essentially a derivative of glycerol, and hydrocarbon chains were conjugated to the glycerol-like site in the structure. The calditol with an ether linkage in the molecule suggested an important role regarding the properties of heat-resistance and acid-resistance observed inSulfolobales. Presented at the international workshop on “Molecular Biology and Biotechnology of Extremophiles and Archaebacteria,” Wako, Japan, August 1993.     

Accumulation of nitrous oxide in aerobic groundwaters

N₂O concentrations in the groundwaters collected in the Kanto District and Nagano Prefecture in Japan and five counties in New York State were determined. These N₂O data were obtained from the water samples from wells, springs and seepages from soils in forests and cropping fields. The N₂O concentrations in all samples greatly exceeded those of atmospheric equilibration. The average concentration of 690 nM N₂O is one order of magnitude larger than that in deep ocean. All groundwaters of the present study were aerobic with a high level of NO₃⁻, but with the absence of NO₂⁻ and NH₄⁺, and with a very low level of dissolved organic carbon. These characteristics suggest that the nitrate respiration in the aquifers is of little significance for the production of N₂O.

ΔN₂O/NO₃⁻ molar ratios in the groundwaters were between 10⁻⁴ and 10⁻² (Δ indicates the excess gas over that which would be in equilibrium). This supports the above view since the observed N₂O yield agrees with that reported for the production during an ammonia oxidation. If nitrification was indeed a major mechanism for the production of groundwater N₂O, subsequent release of N₂O from the aquifers that are polluted with nitrogen may deserve more close attention as a potential source of atmospheric N₂O via diffusion and discharge.     

Effect of pitting corrosion on strength of web plates subjected to patch loading

Pitting corrosion is typical corrosion observed on coated hold frames in way of cargo holds of bulk carriers, which exclusively carry coal and iron ore. Extensive survey on the effect of pitting corrosion on structural strength under a wide variety of loading conditions is necessary to clarify the relationship between pitting intensity and residual strength in detail. In the present study, a series of tests has been conducted on structural models which consist of web, shell and face plates to investigate the effect of pitting corrosion on strength of web plates subjected to patch loading. In these tests, artificial pitting was made on the web plates and two equal concentrated loads have been applied vertically at the one third points of simply supported models. It was found that web crippling behavior is strongly affected by the pit distribution on the web plates. According to the FE-analyses following the experiment, ultimate strength of the web plates with pitting under patch loading is a little smaller than or almost the same as that of the web plates with uniform corrosion in terms of average thickness loss.