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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.

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.

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.

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.

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.

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.

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.

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.

In this study, we experimentally and theoretically show that the intensities of bright spots in a spherical aberration (C(s))-uncorrected high-angle annular dark-field (HAADF) scanning transmission electron microscope (STEM) image of [011]-oriented Co(3)O(4), which has two different numbers of Co atoms in the projected atomic columns, are reversed with increasing sample thickness. However, C(s)-corrected HAADF STEM images produce intensities that correctly depend on the average number of atoms in the projected atomic columns. From an analysis based on the Bloch-wave theorem, it is found that an insufficient semiangle of the incident convergent beam yields intensities that do not depend on the average atomic number in the atomic columns.

A simple and robust method to precisely determine local strain fields using energy-unfiltered convergent-beam electron diffraction is presented. This method involves the subtraction of background intensity, the extraction of higher-order Laue-zone lines by tracing using a Radon transformation and a system of analytical strain determination without the need for an optimization routine such as chi2-based minimization. As an example, the measurement of residual strain in a silicon-on-insulator wafer is demonstrated. It is found from micro-Raman spectroscopy analysis that, at the nanometre scale, this measurement succeeds with an accuracy of 0.06%.

While the bulk of cellular mRNA is known to be exported by the TAP pathway, export of specific subsets of cellular mRNAs may rely on chromosome region maintenance 1 (CRM1). One line of evidence supporting this hypothesis comes from the study of mRNAs of certain early response genes (ERGs) containing the adenylate uridylate-rich element (ARE) in their 3' untranslated regions (3' UTRs). It was reported that HuR-mediated nuclear export of these mRNAs was CRM1-dependent under certain stress conditions. To further examine potential CRM1 pathways for other cellular mRNAs under stress conditions, the nuclear export of human interferon-alpha1 (IFN-alpha1) mRNA, an ERG mRNA induced upon viral infection, was studied. Overproduction of human immunodeficiency virus type 1 Rev protein reduced the expression level of the co-transfected IFN-alpha1 gene. This inhibitory effect, resulting from nuclear retention of IFN-alpha1 mRNA, was reversed when rev had a point mutation that made its nuclear export signal unable to associate with CRM1. Leptomycin B sensitivity experiments revealed that the cytoplasmic expression of IFN-alpha1 mRNA was arrested upon inhibition of CRM1. This finding was further supported by overexpression of DeltaCAN, a defective form of the nucleoporin Nup214/CAN that inhibits CRM1 in a dominant-negative manner, which resulted in the effective inhibition of IFN-alpha1 gene expression. Subsequent RNA fluorescence in situ hybridisation and immunocytochemistry demonstrated that the IFN-alpha1 mRNA was colocalised with CRM1, but not with TAP, in the nucleus. These results therefore imply that the nuclear export of IFN-alpha1 mRNA is mediated by CRM1. However, truncation of the 3' UTR did not negatively affect the nuclear export of IFN-alpha1 mRNA that lacked the ARE, unexpectedly indicating that this CRM1-dependent mRNA export may not be mediated via the ARE.

Atomic resolution low-angle bright-field (LABF) scanning transmission electron-microscope (STEM) images and high-angle bright-field (HABF) STEM images of [011]-orientated Si have been experimentally obtained together with high-angle annular dark-field (HAADF) STEM images. The contrast formation mechanisms of the LABF STEM and HABF STEM images are examined in comparison with HAADF STEM images. The HABF STEM images independent of defocus and thickness have spatial resolution comparable with HAADF STEM images, and are shown to be given as a simple convolution under the non-dispersion approximation of localized Bloch waves.

A model of herpes simplex virus type 1 (HSV-1) infection was developed in rats to study systemic immune responses elicited by intravitreous inoculation of the virus. HSV-1 inoculation led to distinct granulocyte-macrophage colony-stimulating factor (GM-CSF)-producing memory T cells, which did not develop in rats inoculated with either HSV-1 intraperitoneally or inactivated HSV-1 intravitreously. On subsequent intraperitoneal viral boosting, systemic GM-CSF production was elicited as a secondary immune response that caused neutroeosinophilia. To examine the role of GM-CSF in anti-herpetic immunity, cytokine-producing and -nonproducing rats were intravitreously challenged with HSV-1, which causes lethal encephalitis. Only intravitreously primed rats were protected upon production of GM-CSF. Furthermore, pretreatment with recombinant GM-CSF protected unimmunized rats against the encephalitis. It is thus strongly suggested that the production of GM-CSF leads to anti-HSV-1 immunity against the transneuronal spread of challenged HSV-1 within the visual system.

The jaw of Chlamydoselachus anguineus, a living archaic fish, were used as materials and the histological structure of jaw, the development of teeth, the eruption, movement and shedding of teeth, and the developmet of dermal teeth were observed by optical microscopy.Formation of the hard tissues of the jaw teeth and the dermal teeth occurs as following order: formation of enamel matrix→formation of predentin and calcification of enamel→formation of dentin and basal layer. The tissues which contribute to these teeth development are both the epithelial and the mesenchymal elements, although dental lamina does not appear in the development of the dermal teeth. The main epithelial element consists of the inner and the outer enamel epithelia, which are derived from the dental lamina, the oral mucous epithelium or the epidermis. The mesenchymal element consists of the dental papilla and its surrounding tissues, which are derived from the lamina propria of the oral mucous membrane or the dermis. Odontoblasts differentiate in the dental papilla and ameloblasts differentiate in the inner enamel epithelium. The enamel matrix is formed on the dental papilla side of the basement membrane, and then the predentin and the dentin are made in the dental papilla. The occeous tissue of the basal layer is formed under the dental papilla by mesenchymal cells. The fibrous connective tissue layer, which surrounds the basal layer of the tooth, contributes to eruption, movement and shedding of the jaw teeth.Because of the histologicaland developmental nature, the dermal teeth and the teeth of Chiamydoselachus and other chondrichthyes can be recognized as the basic type of the whole vertebrate exoskeletons.

The inner surface of the mouth cavity as far as the beginning of the alimentary canal was examined in young specimens of this Salmonidae fish. Using radiographic techniques and after examination of unstained and alizarin red stained preparations, two groups of teeth were found in the pharynx of this fish. One group teeth was located on the two pairs of the upper pharyngeal bones and also teeth were found on the single pair of the lower pharyngeal bones.The other group consisted of very small teeth situated on the “gill raker bones”, which are very small bones attached to the pharyngeal surface of the branchial arch skeleton. Histologically, both groups of teeth were found to have similar features to oral teeth. In other words, they are polyhyodont, and are attached via bony pedicles to the basal bone, having homogeneous dintin without any dentinal tubules.However, in the development of these teeth, only the teeth on the gill raker bones have no dental lamina. According to the literature, “thorn-like” structures are present on the gill-rakers of many teleostei. Our observations show that the structures on the “gill raker” of Salmo gaird neri irideus are true teeth, and since they could be described as having a “thorn-like” appearance, we would like to point out that the “thorn-like” structures of other teleostei, referred to in the literature, might also be teeth indeed.

サケ科魚類の口腔に歯が多数分布していることはよく知られているが, 著者らはニジマスでは咽頭にも咽頭歯と鰓耙骨歯 (鰓弓面の鰓耙にあり, 鰓耙骨に植立する) の2群の歯があることをみて報告してきた。ここでは, このような歯が他のサケ科3属5種 (サケ属ヒメマスとヤマメ, ニジマス属ブラウンマス, イワナ属イワナとカワマス) にも分布しているかどうかを検索した。口腔から消化管起始部まで, X線撮影やalizarin red染色等を行って内腔面を精査し, また光顕組織標本を作製して鏡検した。その結果, 咽頭に上下の咽頭歯群と多数の鰓耙骨歯が認められた。これらの歯は口腔の歯と同様, 円錐形で均質な象牙質からなり, 多生歯性で, ただ微細な鰓耙骨歯だけは歯堤の出現なしに形成されるなど, ニジマスと同様であった。結論として, これらの淡水産マス類では口腔と咽頭に同様の歯が共通してみられ, 分布状態もほとんど同様であるが属により若干差異がみられた。

The mercuration and thallation of benzene and substituted benzenes was studied with mercuric and thallic trifluoroacetate, respectively, in trifluoroacetic acid. With the shortest reaction time (1 sec) at 0°, the relative rate of mercuration of toluene compared to that of benzene was 17.5, with the isomer distribution in toluene of: ortho , 17.4%; meta , 5.9%; and para , 76.7%. The isomer distribution in toluene varied with the reaction time, significantly more at 25° than at 0°. The competitive thallation of benzene and toluene with thallic trifluoroacetate in trifluoroacetic acid at 15° showed the relative rate, toluene/benzene, to be 33, with the isomer distribution in toluene of: ortho , 9.5%; meta , 5.5%; and para , 85.0%. With increasingly higher reaction temperatures in both mercuration and thallation reactions of aromatics, isomerization (both intramolecular and intermolecular) within the relevant ortho - and para -metallated intermediate ions and/or of the isomers becomes more important. Competitive rates and isomer distributions of mercuration and thallation of benzene and substituted benzenes were also determined. The predominant para substitution in both mercuration and thallation of methylbenzenes reflects, besides some steric factors, the strong stabilizing effect of para methyl groups on the arenium ion intermediates. Under predominantly kinetically controlled conditions, no anomalous increase in the amount of meta substitution was observed.

When homosulf amine was used through root canal therapy, it has been known that the germicidal effect was reduced by anti-agents produced in the root canals. Among these anti-agents were necrotic tissues, pus, secretion and blood etc. In this experiment, therefore, it was tested how these unfavorable agents worked in vitro upon the germicidall effect of this medicine.The chief results of this research are as follows:1) Without anti-agents, the local use of 5% homosulfamine solution would be considered most effective in vitro.2) By consideration of the time of 50% LD. of all amount of bacteria, the effect of 20% homosulfamine solution was remarkably reduced by pus only to show that of 5% solution.3) 5% Homosulfamine solution was reduced by secretion or gelatine to 1.25% solution in the effect.4) No change caused by serum was observed.5) By using urea with homosulfamine, the reduction of the effect by pus could be abated.

The changes in monoamine levels of different brain regions following Japanese encephalitis virus (JEV) intraperitoneal inoculation were examined in experimentally JEV-infected mice. In addition, virus distribution was studied using infectivity assay and immuno-histochemistry of viral antigen. 1) The level of monoamines in brain tissues was not affected by 48 hours after viral inoculation, but marked effects were elicited at 96 hours after the inoculation. The cerebral concentration of 5-hydroxyindole-3-acetic acid (5 HIAA) was increased, while that of dopamine (DA) showed a decrease. Especially these alteration were observed in the cerebral cortex, but not in the cerebellum. 2) The viral growth in the brain was observed at 48 hours after the inoculation. The growth in the cerebellum, however, was found to be lower than those in other cerebral regions. 3) The viral antigen was detected in the cerebral cortex, hippocampus, mesencephalon and diencephalon in addition to the substantia nigra and striatum. From these results, it is presumed that clinical manifestation of JEV infection may involve the changes in the metabolism of neurotransmitter, especially those of DA and serotonin in the brain.

The AlCl3-catalyzed acylation of benzene with 2-methylbutanedioic anhydride afforded a mixture of 3-benzoyl-2-methylpropanoic acid and 3-benzoylbutanoic acid. The intramolecular acylation of 2-benzylbutanedioic anhydride in the presence of AlCl3 gave a mixture of 4-oxo-1,2,3,4-tetrahydro-2-naphthoic acid and 3-oxo-2-indanacetic acid. The results were discussed in terms of the solvent effect on the acylations.

For the growth of NdAl3(BO3)4 crystals, BaB4O7 was chosen as a flux on the basis of the guide proposed by the authors.NdAl3(BO3)4 crystals were grown by slow cooling of BaB4O7 flux in which Nd2O3, Al2O3and B2O3 were dissolved as solutes. The growth was conducted by heating mixtures at 1150°C for 10 h followed by cooling to 600°C at a rate of 5°C/h. Crystals grown in BaB4O7 flux were hexagonal rod in shape, up to 7.1×1.5×0.7 mm in size, bluish purple in color and transparent. It was shown that the flux chosen was suitable for the growth of objective crystals.

The AlCl3-catalyzed acylation of benzene with 2-phenylbutanedioic anhydride was found to give a mixture of 3-benzoyl-2-phenylpropanoic acid, isomeric 3-benzoyl-3-pehnylpropanoic acid, and 3-oxoindan-1-carboxylic acid. The distribution of the products was affected markedly by the amount of AlCl3 or solvents used.