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Thready stripe-polyacrylamide (PAAm) pattern was fabricated on a thermoresponsive poly(N-isopropylacrylamide) (PIPAAm) surface, and their surface properties were characterized. A PIPAAm surface spin-coated with positive photoresist was irradiated through a 5 µm/5 µm or a 10 µm/10-µm black and white striped photomask, resulting in the radical polymerization of AAm on the photoirradiated area. After staining with Alexa488 bovine serum albumin, the stripe-patterned surface was clearly observed and the patterned surface was also observed by a phase contrast image of an atomic force microscope. NIH-3T3 (3T3) single cells were able to be cultured at 37°C on the patterned surfaces as well as on a PIPAAm surface without pattern, and the detachment of adhered cells was more rapidly from the patterned surface after reducing temperature. Furthermore, the rate of detachment of 3T3 confluent cell sheet on the patterned surface was accelerated, compared with on a conventional PIPAAm surface under the static condition. The rate control of cell sheet recovery should contribute the preservations of cell phenotype and biological functions of cell sheet for applying to clinical trials.

Aim Tumor necrosis factor-α (TNF-α) is a pleiotropic cytokine involved in various inflammatory diseases. The only production of TNF-α in the liver is thought to be from hepatic macrophages known as Kupffer cells, predominantly in response to bacterial lipopolysaccharide (LPS). Methods Primary cultured rat hepatocytes were used to analyze TNF-α expression in response to the pro-inflammatory cytokine, interleukin-1β (IL-1β). Livers of rats subjected to LPS-induced endotoxemia were analyzed. Results Immunocytochemistry and enzyme-linked immunosorbent assays demonstrated that IL-1β-treated rat hepatocytes secreted TNF-α, and RNA analyses indicated that TNF-α mRNA was induced specifically by IL-1β. Northern blot analysis showed that not only mRNA, but also a natural antisense transcript (asRNA), was transcribed from the rat Tnf gene in IL-1β-treated hepatocytes. TNF-α was detected in the hepatocytes of LPS-treated rats. Both TNF-α mRNA and asRNA were expressed in the hepatocytes of LPS-treated rats, human hepatocellular carcinoma and human monocyte/macrophage cells. To disrupt the interaction between TNF-α asRNA and TNF-α mRNA, sense oligonucleotides corresponding to TNF-α mRNA were introduced into rat hepatocytes resulting in significantly increased levels of TNF-α mRNA. One of these sense oligonucleotides increased a half-life of TNF-α mRNA, suggesting that the TNF-α asRNA may reduce the stability of TNF-α mRNA. Conclusion IL-1β-stimulated rat hepatocytes are a newly identified source of TNF-α in the liver. TNF-α mRNA and asRNA are expressed in rats and humans, and the TNF-α asRNA reduces the stability of the TNF-α mRNA. Hepatocytes and TNF-α asRNA may be therapeutic targets to regulate levels of TNF-α mRNA.

Nonlocality in spherical-aberration-corrected high-angle annular dark-field (HAADF) scanning transmission electron microscope (STEM) images is theoretically and experimentally examined using the absorption potential describing thermal diffuse scattering (TDS). A detailed comparison between the simulated and the experimentally obtained high-quality HAADF STEM images of an Si(110) bulk structure and a PbTiO3(100)/SrTiO3(100) interfacial structure unambiguously demonstrates the need to use a nonlocal TDS absorption potential. The nonlocality in the TDS absorption potential cannot be ignored in a detailed analysis of spherical-aberration-corrected HAADF STEM images of materials consisting of several heavy elements, although it can be completely disregarded for those consisting of only light elements.

Antisense transcription is a widespread phenomenon in the mammalian genome and is believed to play a role in regulating gene expression. However, the exact functional significance of antisense transcription is largely unknown. Here, we show that natural antisense (AS) RNA is an important modulator of interferon-α1 (IFN-α1) mRNA levels. A ~4-kb, spliced IFN-α1 AS RNA targets a single-stranded region within a conserved secondary structure element of the IFN-α1 mRNA, an element which was previously reported to function as the nuclear export element. Following infection of human Namalwa lymphocytes with Sendai virus or infection of guinea pig 104C1 fetal fibroblasts with influenza virus A/PR/8/34, expression of IFN-α1 AS RNA becomes elevated. This elevated expression results in increased IFN-α1 mRNA stability because of the cytoplasmic (but not nuclear) interaction of the AS RNA with the mRNA at the single-stranded region. This results in increased IFN-α protein production. The silencing of IFN-α1 AS RNA by sense oligonucleotides or over-expression of antisense oligoribonucleotides, which were both designed from the target region, confirmed the critical role of the AS RNA in the post-transcriptional regulation of IFN-α1 mRNA levels. This AS RNA stabilization effect is caused by the prevention of the microRNA (miRNA)-induced destabilization of IFN-α1 mRNA due to masking of the miR-1270 binding site. This discovery not only reveals a regulatory pathway for controlling IFN-α1 gene expression during the host innate immune response against virus infection but also suggests a reason for the large number of overlapping complementary transcripts with previously unknown function. Electronic supplementary material The online version of this article (doi:10.1007/s00018-012-1216-x) contains supplementary material, which is available to authorized users.

This research focused on both the psychological benefits and costs of crying. We investigated the relationships of intrapersonal and interpersonal consequences of crying. Female nurses (N = 300) were requested to describe one of the most impressive negative episodes where they had cried. Then, they were asked to complete a questionnaire including a scale of their psychological changes after the crying episode and the social reactions when they cried. Factor analysis revealed five components of the psychological changes scale. Solitary crying had greater effects for both psychological benefits and costs after crying than crying in front of others. Factor analysis revealed three components of the scale of social reactions. When they cried in front of others, "catharsis", "positive attitude", and "recognition of the relationship with others" after crying were associated with "empathy and social support" from others. The factors of "recognition of negative reality" and "negative attitude" were associated with "criticism and slander" from others. These results were discussed in terms of the communicative functions and the reflective functions of adult crying.

Esterification of several types of mono- and disubstituted phenols with various mono- and dialkanoyl chlorides was performed in phase-transfer catalysis conditions, using tetrabutylammonium chloride in a mixture of aqueous NaOH and dichloromethane. The process is particularly efficient (almost quantitative yields) as well as rapid (only 5 min reaction time, at a temperature of 0 °C).

The human immunodeficiency virus type 1 (HIV-1) regulatory protein, Rev, mediates the nuclear export of unspliced gag and singly spliced env mRNAs by bridging viral RNA and the export receptor, CRM1. Recently, rat CRM1 was found to be less efficient than human CRM1 in supporting Rev function in rats. In this study, to understand the role of CRM1 in HIV propagation, the mechanism underlying the function of human and rat CRM1 in HIV-1 replication was investigated in rat cells. The production of viral particles, represented by the p24 Gag protein, was greatly enhanced by hCRM1 expression in rat cells; however, this effect was not simply because of the enhanced export of gag mRNA. The translation initiation rate of gag mRNA was not increased, nor was the Gag protein stabilized in the presence of hCRM1. However, the processing of the p55 Gag precursor and the release of viral particles were facilitated. These results indicated that hCRM1 exports gag mRNA to the cytoplasm, not only more efficiently than rCRM1 but also correctly, leading to efficient processing of Gag proteins and particle formation.

Various thioesters were obtained through an efficient phase-transfer catalysis method, by treating several thiophenols with different acyl chlorides, in a biphasic system composed of 10% aqueous NaOH and dichloromethane in the presence of tetrabutylammonium chloride. The thiolation reaction was complete in only 5 minutes, at 0°C.

A nitrogen balance study was carried out by setting up a test paddy in order to estimate the nitrogen pollution load potential (NPLP) from farmland in the Tedori River Alluvial Fan Areas and the load from the entire area under consideration was estimated using the cropping record and fertilizer application rate (FAR). The total NPLP was estimated to be 261 tons/year and the load from the paddy 79 tons/year for 5,704 ha, which would translate to an intensity of 14 kg/ha, while the load from the vegetable field was estimated to be 118 tons/year for 215 ha, which would equate to an intensity of 549 kg/ha. The pollution loads for the vegetables were significantly greater than those for the rice. The load for the barley was 57 tons/year for 261 ha (216 kg/ha) and that for the orchards was 23 tons/year for 64 ha (359 kg/ha). The estimation of soybean load was a negative 15 tons/year for 717 ha (−21 kg/ha), which meant that the nitrogen in the yield was greater than the FAR. The results also confirmed the yield absorption ratio in relation to the FAR. The load from the paddy for the entire area was also estimated using the percolation rate and the water quality load underneath the farmland was estimated to be 89 tons/year.

We investigated the use of irrigation water for a successive planting of watermelon followed by Japanese radish in a sand dune area in the Hokuriku Region. The main results were as follows: (1) Upland field tested in the study used a large volume of irrigation water in spite of its location in the Hokuriku Region where rainfall was plentiful. Notwithstanding this, frequent irrigation was considered necessary. (2) Watermelon was cultivated as a Spring–Summer crop with a mean cultivation period of 102 days, during which it was irrigated for 59 days at 1.7-day average intervals with an irrigation volume of 7.1 mm/watering and a total irrigation volume of 428 mm over its entire growing season. The amount of rainfall during the period was 604 mm during 42 days of rainfall. Soil moisture levels in the upland field were kept at relatively high values ranging between pF1.3 and pF1.5 at 15-cm depth with frequent irrigation during no-rainfall period. (3) Japanese radish was cultivated as a Summer–Autumn crop with a mean cultivation period of 71 days during which it was irrigated for 39 days at 1.9-day average intervals with an irrigation volume of 9.4 mm/watering and a total irrigation volume of 358 mm over its entire growing season. The amount of rainfall during the period was 376 mm during 29 days of rainfall. Frequent irrigations were used to maintain the soil moisture levels in the upland field within the range of pF1.4–pF1.6 at 15 cm depth. (4) During the irrigation period, the amount of water used for irrigation was 5.9 mm/day in watermelon and 6.1 mm/day in Japanese radish which are greater than evapotranspiration (ET). In addition, during pre-irrigation, the preparation of the seeding bed can be carried out at the right time which contributes to producing high-quality Japanese radish in adequate quantity. In addition, pre-irrigation for preparation of bed for watermelon transplanting is very important for better time management and high-quality production. For this purpose, the irrigation facilities need to be functioning very well.

The effect of the chromatic aberration (C(c)) coefficient in a spherical aberration (C(s))- corrected electromagnetic lens on high-resolution high-angle annular dark field (HAADF) scanning transmission electron microscope (STEM) images is explored in detail. A new method for precise determination of the C(c) coefficient is demonstrated, requiring measurement of an atomic-resolution one-frame through-focal HAADF STEM image. This method is robust with respect to instrumental drift, sample thickness, all lens parameters except C(c), and experimental noise. It is also demonstrated that semi-quantitative structural analysis on the nanometer scale can be achieved by comparing experimental C(s)- corrected HAADF STEM images with their corresponding simulated images when the effects of the C(c) coefficient and spatial incoherence are included.

Human immunodeficiency virus type 1 Rev (regulator of the expression of the virion) protein was shown to reduce the expression level of the co-transfected luciferase reporter gene (luc+) introduced to monitor transfection efficiency. We studied the mechanism of the inhibitory Rev effect. The effect, caused by nuclear retention of luc+ mRNA, was reversed if rev had a point mutation that makes its nuclear export signal (NES) unable to associate with cellular transport factors. The Rev NES receptor CRM1 (chromosome region maintenance 1)-specific inhibitor, leptomycin B, blocked luc+ mRNA export. This finding was also supported by the overexpression of delta CAN, another specific CRM1 inhibitor that caused inhibition of luciferase gene expression. Experiments involving tsBN2 cells, which have a temperature-sensitive RCC1 (regulator of chromosome condensation 1) allele, demonstrated that luc+ expression required generation of the GTP-bound form of RanGTPase (RanGTP) by RCC1. The constitutive transport element (CTE)-mediated nuclear export of luc+ mRNA was found to also depend upon RanGTP. Nuclear export of luc+ mRNA is thus suggested to involve CRM1 and RanGTP, which Rev employs to transport viral mRNA. The Rev effect is therefore considered to involve competition between two molecules for common transport factors.

This research proposes an assessment for nitrogen (N) pollution loads potential from farmland, based on comparison of N in objective crop yields with standard fertilizer usage. N in the objective yield was calculated using the “Standard Table of Food Composition in Japan”. Three findings were made by considering yields removed from farmland. First, paddy rice and beans have a low pollution potential, with rice paddies showing a negative pollution potential for N (around −14 kg/ha). Second, almost all vegetable and orchard crops tested had a high pollution potential for N, although this differed from crop to crop. Third, our outcomes align well with farmland pollution potential and non-absorbed nitrogen (NAN) as defined by Nishio, although the latter and indices rely on a laborious and complicated method. The correlation coefficients were 0.745 (R 2 = 0.555). These outcomes show the effectiveness of our proposed assessment for potential environmental pollution loads.

A many-beam dynamical theory for plan-view high-resolution transmission electron microscopy (HRTEM) images of multilayer systems without the limitation of a superlattice cell is proposed. The accuracy of our method is examined by comparing convergent-beam electron-diffraction calculations of Si(011) and HRTEM calculations of a system of epitaxial Al(100) on GaAs(100). Furthermore, this method is applied to CdSe clusters embedded in MgO, where it is revealed that the relative shift of their crystal-lattice planes produces moiré-like fringes.

A polar-dependent phase with spontaneous atomic ordering is found in a nonpolar (1120) plane In0.08Ga0.92N film grown by metalorganic vapor-phase epitaxy. An atomic arrangement is a periodic sequence of group-III sublattices, such as In0.04Ga0.96N/In0.12Ga0.88N, that is only observed in a nitrogen polar region through systematic transmission electron microscopy investigation. Cathodoluminescence (CL) in the nitrogen polar region, i.e., spontaneously ordered atomic structure of InGaN, reveals anomalous emission behavior, specifically an S-shape-like (increase–decrease) temperature dependence of CL peak energy. It is suggested that the spontaneous ordered atomic structure of InGaN plays the role of a localized center owing to band gap shrinkage, which has been reported in other III–V alloy systems.

The structure and composition of the 1/4{1 1 0} twin boundary in α - Zn 7 Sb 2 O 12 have been determined by using quantitative high-angle annular dark field scanning transmission electron microscopy (HAADF STEM) analysis. The noise in the experimental HAADF STEM images is reduced by using the maximum entropy method and average processing, and the parameters used in dynamical simulations are experimentally determined. From the analysis, it has been found that octahedral sites in the twin boundary slightly shift parallel to the [1 1 0] direction, and a reduction of the Sb concentration at the octahedral sites on the plane adjacent to the twin boundary was detected. The reduction was measured from three regions in the same twin boundary, and the Sb concentrations were 4 ± 3 , 8 ± 3 and 19 ± 2 at % from 33 at%.

The quantitative measurement of a crystal bending effect is performed using low-order zone-axis convergent beam electron diffraction (CBED) patterns. Although the accuracy of the present method is inferior to that of the method of using split higher order Laue zone lines, this method enables us to estimate the crystal bending effect at a region very close to the interface and to easily judge whether the crystal bending effect results in a tensile bend or a compressive bend. As an application of the present method, the crystal bending effect at a region close to the SiGe/Si interface was measured. It was found that the crystal bending effect is due to a thin-foil relaxation of almost 0.3 degrees at a region that is approximately 10 nm away from the interface.

Pinacol-type coupling reaction products presenting a high meso-diastereoselectivity (the ratio dl/meso was 4/96 up to 1/99) were obtained in fair to good yields (24–69%) using several aromatic aldehydes as starting materials and aluminium powder/copper sulfate as catalysts, in water, under reflux conditions.

Kahokugata Lake, a closed lake, has been subject to eutrophication. This research was conducted to clarify the actual phenomena and evaluation of the discharges of N and P from paddy test fields in the lowlands into Kahokugata Lake. A comprehensive mass balance of N and P was obtained from 4 years of study. About N, the mean value of harvested unhulled rice (79.9 kg/ha) was greater than mean controlled release fertilizer inputs (56.7 kg/ha). Other inputs and outputs include N in atmospheric acid deposition (21.4 kg/ha) and N fixation–denitrification (9.2 kg/ha). The rice straw recycled after harvest was balanced by straw produced in the succeeding year. The runoff and percolation losses discharged into the lake was 11.3 kg/ha, (8.6% of total inputs). Since the rice harvested was consumed domestically, which taking out from the farmland and, therefore, nitrogen pollutant into the lake was becoming small, paddy rice at this site is considered an “anti-polluting, purifying or cleansing” crop. The P content in harvested rice (39.4 kg/ha) was balanced by fertilizer inputs (36.4 kg/ha). Previous studies examining inflow–outflow relationships without considering a comprehensive mass balance may lead to erroneous conclusions. Our findings indicate paddy rice in lowlands could be an environmentally friendly crop and can play an important role in reducing pollution of lakes, and therefore should be considered in land use planning.

The shape effect of microtwins on high-resolution transmission electron microscopy (HRTEM) images is demonstrated with the help of a detailed comparison between experimental HRTEM images of recrystallized silicon (including many microtwins) and corresponding dynamical simulations. The analysis points to an important fact: The shape effect of microtwins leads to artificial contrasts. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

An extended method is proposed for the precise simulation of high-angle annular dark-field (HAADF) scanning transmission electron-microscope (STEM) images for materials containing elements with large atomic numbers and for thick specimens. The approach combines a previously reported method utilizing two kinds of optical potential [Watanabe, Yamazaki, HashimotoShiojiri (2001). Phys. Rev. B, 64, 115432] with a representation of a crystal sliced into multiple layers. The validity of the method is demonstrated by simulated images for elements with the diamond structure (Si, Ge and alpha-Sn) and for the perovskite BaTiO3.

Structural analysis of the submicron area of an InGaN thin film was conducted using transmission electron microscopy. By obtaining the selected area diffraction patterns of c -plane InGaN/GaN and a -plane InGaN/GaN films, the local lattice parameters were measured. Coherent growths were checked at the respective samples for different growth orientations, and the In concentration of the InGaN layer was also measured. Furthermore, there is a slight tilt in the growth direction between the GaN and InGaN layers in the a -plane InGaN/GaN film. (© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

A systematic structural analysis of a-plane InGaN/GaN was performed using electron diffraction, high-resolution transmission electron microscopy (HRTEM), and energy-dispersive X-ray spectroscopy. From the results of local structural analysis using Fourier diffractograms of HRTEM images, it was found that a-plane InGaN/GaN exhibits coherent epitaxial growth and a slight tilt of the growth direction between the GaN and InGaN layers. Furthermore, the In concentration and tilt angle increase simultaneously with increasing distance from the InGaN/GaN interface.

Nanoparticles found in interlayers between the grains of Bi2O3-doped ZnO were analyzed using electron diffraction, high-resolution transmission electron microscopy and energy-dispersive X-ray spectroscopy. Both ZnO multiple twinned particles (MTPs) and single nanoparticles of less than 5 nm in diameter were found in the Bi-poor interlayers. The MTPs with the wurtzite structure give rise to no defects or structural changes in the present size because the angular misgap of five-fold symmetry is very small. Furthermore, β-Bi2O3 nanoparticles reaching a few tens of nanometers in diameter were formed in the Bi-rich interlayers.

Reductions in bright-field (BF) scanning transmission electron microscopy (STEM) and high-angle annular dark-field (HAADF) STEM image calculations with the aid of Bloch wave symmetry are discussed under assumptions that an absorption potential is written by a local potential and a zero-order Laue zone lies parallel to the crystal surface. Translational symmetry allows us to take only partial incident beams in the first Brillouin zone instead of enormous number of partial incident beams in a large convergent disk. Two dimensional point group confines partial incident beams to an irreducible area in addition to factoring a dispersion matrix into noninteracting submatrices on a high symmetry line using the projection operator. The drastic reductions in computing time and memory enable us to readily calculate various BF STEM and HAADF STEM images. The validity and accuracy are demonstrated in comparisons with high resolution experimental BF STEM and HAADF STEM images.

Atomic-resolution incoherent high-angle annular dark field (HAADF) scanning transmission electron microscopy (STEM) images of [011]-oriented Si have been recorded using different incident beam probes, and analysed by means of dynamical image calculation based on the Bloch wave description. It is shown how atomic-resolution images are influenced by the semiangle of the probe and the spherical aberration and defocus of the probe-forming lens. The resolution of an incoherent HAADF STEM image can be simply perceived by the contrast transfer function of incoherent imaging which is the Fourier transform of the incident probe intensity.

Using a slightly tilted convergent electron beam, high-angle annular dark field scanning transmission electron microscopy observations have been performed of a [0 11]-oriented Si crystal. A small tilt of the crystal zone axis with respect to the coma-axis of the probe-forming lens causes a difference in intensity between bright spots of a Si dumbbell. The semiangle of the beam probe and the tilting angle with respect to the specimen hormal were determined by means of convergent beam micro-diffraction. The simulation using these parameters accounts for the image contrasts satisfactorily.

Artificial bright spots, which appear in some atomic resolution high-angle annular dark field scanning transmission electron microscope (HAADF STEM) images, have been accounted for by simulations based on Bloch wave description. This is illustrated with Si and SrTiO 3 images. The simulation reveals that bright spots on no-atomic columns in [011]-orientated Si images are produced by thermal diffuse scattering from Si atoms on their surrounding atomic columns, which are under the subsidiary peaks in the incident convergent electron probe. Similarly, bright spots on oxygen columns in [001]-orientated SrTiO 3 images are ascribed to Sr and Ti atoms in their surrounding atomic columns rather than O atoms in the O columns. The probe function, therefore, provides a simple explanation for the appearance of these artificial spots.

Background A hallmark of HIV-1 gene expression is that unspliced genomic RNA, which also acts as mRNA for the expression of Gag/Pol, is exported to the cytoplasm. Rev directs this transport through the nuclear export signal (NES). Results Fluorescence in situ hybridization and immunocytochemistry demonstrated that gag mRNA, Rev, and its NES receptor, CRM1, and RanGTPase formed nuclear tracks which were congruent with underlying β-actin bundles. Actin bundle formation was confirmed electron-microscopically. These bundles were observed upon Rev-containing gag RNP formation. The loss of bundles was associated with the nuclear retention of gag mRNA. Reverse transcription-polymerase chain reaction analysis of both cytoplasmic and nuclear gag mRNAs demonstrated that disruption of nuclear actin filament formation by latrunculin-B (LAT-B), an F-actin depolymerizing compound, resulted in the dose-dependent inhibition of gag mRNA export. The differential subtyping of the mRNA-positive cells confirmed morphologically the effect of LAT-B treatment. The export inhibition was specific to gag mRNA and export of fully spliced HIV-1 tat/rev mRNAs as well as cellular GAPDH mRNA was not affected by the compound. Conclusions Nuclear β-actin bundles are suggested to be functionally involved in the Rev-dependent nucleocytoplasmic transport of intron-containing HIV-1 gag mRNA.

A distortion measurement in a region close to the interface between different materials in LSI is performed using a convergent beam electron diffraction (CBED) pattern. Split higher-order Laue zone (HOLZ) lines emerge in the CBED pattern so that a stressing region is observed close to the interface. The calculation method of the split HOLZ lines based on kinematical approximation with the sample's deformation model well reflects the experimental results. As a result of split HOLZ line analysis using the present method, it is found that there is distortion depending on the external form of a multi-finger transistor.

Reduction of imines was performed with zinc powder in 5% aq NaOH solution without any organic solvents under mild conditions, and the corresponding amines were obtained in good yields.

Human immunodeficiency virus type 1 (HIV1) Rev has been reported to act by inducing the nucleocytoplasmic transport of unspliced and singly spliced RNAs that encode viral structural proteins. However, our initial experiments indicated the cytoplasmic expression of intron-containing mRNAs in Rev-cells [12]. To determine whether or not the post-transcriptional induction of gene expression by Rev extends to the level of increasing the efficiency of translation, Rev+ and Rev-cells were morphologically examined by means of in situ hybridization and immunofluorescence assays. Hybridization to an intron-specific probe revealed that the transfection of HeLa cells with a rev-defective HIV-1 expression plasmid caused the export of overexpressed, unspliced gag mRNAs, possibly through a default process of nuclear retention. However, subsequent immunofluorescence assaying demonstrated that mRNA exported by Rev, but not mRNA directed through the default process, was translated into the corresponding protein. The findings were extended to a group of singly spliced viral mRNAs that produce Env in the following biochemical analyses. These findings suggest that Rev is directly or indirectly involved in the translational regulation of HIV-1 structural gene mRNAs. We discuss the possibility that Rev could cause the intron-containing transcripts to follow the appropriate pathway to reach the translational machinery.

SmI 2 /HMPA-mediated double reductive coupling of N , N ′-( m -xylylidene)dianiline affords 1,2,9,10-tetrakis( N -phenylamino)[2.2]metacyclophane in good yield.

Thready stripe-polyacrylamide (PAAm) pattern was fabricated on a thermoresponsive poly(N-isopropylacrylamide) (PIPAAm) surface, and their surface properties were characterized. A PIPAAm surface spin-coated with positive photoresist was irradiated through a 5 µm/5 µm or a 10 µm/10-µm black and white striped photomask, resulting in the radical polymerization of AAm on the photoirradiated area. After staining with Alexa488 bovine serum albumin, the stripe-patterned surface was clearly observed and the patterned surface was also observed by a phase contrast image of an atomic force microscope. NIH-3T3 (3T3) single cells were able to be cultured at 37°C on the patterned surfaces as well as on a PIPAAm surface without pattern, and the detachment of adhered cells was more rapidly from the patterned surface after reducing temperature. Furthermore, the rate of detachment of 3T3 confluent cell sheet on the patterned surface was accelerated, compared with on a conventional PIPAAm surface under the static condition. The rate control of cell sheet recovery should contribute the preservations of cell phenotype and biological functions of cell sheet for applying to clinical trials.

The Friedel-Crafts acylations of various aromatic compounds with cyclic anhydrides such as 2-(p-substituted phenyl)butanedioic, 3-phenylpentanedioic and homophathlic anhydrides were carried out under various conditions in order to obtain informations about the regioselectivity of the ring opening of the cyclic anhydrides and about the possible reaction pathways in the acylations.

From the Dirac equation with a periodic scalar potential, an n-beam dynamical formula for the matrix representation of high-energy electron diffraction by a crystal is developed. By combining this with the layer-doubling method, the diffraction from an assembly of crystal slabs having different structures and thicknesses can be evaluated. Dynamical calculations of aluminium, copper and gold at several accelerating voltages have been carried out in a completely parallel manner by the present method and the Bethe method derived from the relativistic Schrodinger equation by replacing corrected mass and wavelength. The relativistic Schrodinger equation is found to be applicable for n-beam dynamical calculations for the Laue case.

Human immunodeficiency virus type-1 (HIV-1) Rev acts by inducing the specific nucleocytoplasmic transport of a class of incompletely spliced RNAs that encodes the viral structural proteins. The transfection of HeLA cells with a rev -defective HIV-1 expression plasmid, however, resulted in the export of overexpressed, intron-containing species of viral RNAs, possibly through a default process of nuclear retention. Thus, this system enabled us to directly compare Rev + and Rev − cells as to the usage of RRE-containing mRNAs by the cellular translational machinery. Biochemical examination of the transfected cells revealed that although significant levels of gag and env mRNAs were detected in both the presence and absence of Rev, efficient production of viral proteins was strictly dependent on the presence of Rev. A fluoroscence in situ hybridisation assay confirmed these findings and provided further evidence that even in the presence of Rev, not all of the viral mRNA was equally translated. At the early phase of RNA export in Rev + cells, gag mRNA was observed throughout both the cytoplasm and nucleoplasm as uniform fine stippling. In addition, the mRNA formed clusters mainly in the perinuclear region, which were not observed in Rev − cells. In the presence of Rev, expression of the gag protein was limited to these perinuclear sites where the mRNA accumulated. Subsequent staining of the cytoskeletal proteins demonstrated that in Rev + cells gag mRNA is colocalized with β-actin in the sites where the RNA formed clusters. In the absence of Rev, in contrast, the gag mRNA failed to associate with the cytoskeletal proteins. These results suggest that in addition to promoting the emergence of intron-containing RNA from the nucleus, Rev plays an important role in the compartmentation of translation by directing RRE-containing mRNAs to the β-actin to form the perinuclear clusters at which the synthesis of viral structural proteins begins.

Tumor necrosis factor-α (TNF-α) is a pleiotropic cytokine involved in various inflammatory diseases. The only production of TNF-α in the liver is thought to be from hepatic macrophages known as Kupffer cells, predominantly in response to bacterial lipopolysaccharide (LPS).Primary cultured rat hepatocytes were used to analyze TNF-α expression in response to the pro-inflammatory cytokine, interleukin-1β (IL-1β). Livers of rats subjected to LPS-induced endotoxemia were analyzed.Immunocytochemistry and enzyme-linked immunosorbent assays demonstrated that IL-1β-treated rat hepatocytes secreted TNF-α, and RNA analyses indicated that TNF-α mRNA was induced specifically by IL-1β. Northern blot analysis showed that not only mRNA, but also a natural antisense transcript (asRNA), was transcribed from the rat Tnf gene in IL-1β-treated hepatocytes. TNF-α was detected in the hepatocytes of LPS-treated rats. Both TNF-α mRNA and asRNA were expressed in the hepatocytes of LPS-treated rats, human hepatocellular carcinoma and human monocyte/macrophage cells. To disrupt the interaction between TNF-α asRNA and TNF-α mRNA, sense oligonucleotides corresponding to TNF-α mRNA were introduced into rat hepatocytes resulting in significantly increased levels of TNF-α mRNA. One of these sense oligonucleotides increased a half-life of TNF-α mRNA, suggesting that the TNF-α asRNA may reduce the stability of TNF-α mRNA.IL-1β-stimulated rat hepatocytes are a newly identified source of TNF-α in the liver. TNF-α mRNA and asRNA are expressed in rats and humans, and the TNF-α asRNA reduces the stability of the TNF-α mRNA. Hepatocytes and TNF-α asRNA may be therapeutic targets to regulate levels of TNF-α mRNA.

The title method is rapid, extremely effective and superior to previously described procedures.

Simultaneous detection of both light and heavy atomic columns is theoretically and experimentally explored with spherical aberration (Cs)-corrected middle-angle bright-field (MABF) scanning transmission electron microscopy (STEM). Optimized MABF STEM visualizes both light O atomic columns and heavy Sr and Ti–O atomic columns for SrTiO3(001) as distinct bright spots and dark spots with characteristic bright rings, respectively, over practical ranges of the probe-forming lens defocus and sample thickness, although medium-heavy Ti–O atomic columns appear as blurred dark spots. The difference in contrast between heavy and light atomic columns is greater than that of annular BF STEM images. The formation of distinctive bright and dark spots is interpreted simply as the difference in the degrees of localization and inelastic absorption of channeling electrons in individual atomic columns by analyses of convergent wave fields inside the crystal in both real and reciprocal space. In addition, Bloch wave expansion of MABF STEM images suggests that bright rings are formed mainly by 2p-like convergent Bloch wave fields localized on heavy atomic columns.