We have studied magnetoresistance and Hall effects for 1.8-nm-thick Pt films grown on a ferrimagnetic insulator Y3Fe5O12 in a wide temperature (0.46-300 K) and magnetic-field (-15-15 T) region. In the low-temperature regime where quantum corrections to conductivity are observed, weak antilocalization behavior observed in Pt films is critically suppressed when the film is attached to Y3Fe5O12. Hall resistance in the Pt film is also affected by Y3Fe5O12, and it exhibits logarithmic temperature dependence in a broad temperature range. The magnetotransport properties in the high-field range are significantly influenced by the interface between Pt and Y3Fe5O12.

We previously reported that multiply-primed rolling circle amplification (MRPCA) using modified random RNA primers can amplify tiny amounts of circular DNA without producing any byproducts. However, contaminating DNA in recombinant Phi29 DNA polymerase adversely affects the outcome of MPRCA, especially for negative controls such as non-template controls. The amplified DNA in negative control casts doubt on the result of DNA amplification. Since Phi29 DNA polymerase has high affinity for both single-strand and double-stranded DNA, some amount of host DNA will always remain in the recombinant polymerase. Here we describe a procedure for preparing Phi29 DNA polymerase which is essentially free of amplifiable DNA. This procedure is realized by a combination of host DNA removal using appropriate salt concentrations, inactivation of amplifiable DNA using ethidium monoazide, and irradiation with visible light from a light-emitting diode lamp. Any remaining DNA, which likely exists as oligonucleotides captured by the Phi29 DNA polymerase, is degraded by the 3'-5' exonuclease activity of the polymerase itself in the presence of trehalose, used as an anti-aggregation reagent. Phi29 DNA polymerase purified by this procedure has little amplifiable DNA, resulting in reproducible amplification of at least ten copies of plasmid DNA without any byproducts and reducing reaction volume. This procedure could aid the amplification of tiny amounts DNA, thereby providing clear evidence of contamination from laboratory environments, tools and reagents.

We report anisotropic magnetoresistance in Pt|Y(3)Fe(5)O(12) bilayers. In spite of Y(3)Fe(5)O(12) being a very good electrical insulator, the resistance of the Pt layer reflects its magnetization direction. The effect persists even when a Cu layer is inserted between Pt and Y(3)Fe(5)O(12), excluding the contribution of induced equilibrium magnetization at the interface. Instead, we show that the effect originates from concerted actions of the direct and inverse spin Hall effects and therefore call it "spin Hall magnetoresistance."

The Inverse Spin-Hall Effect (ISHE), Conversion of Spin Currents into Charge Currents, Has Recently Been Observed in Paramagnetic Metal/Ferrimagnetic Insulator Bilayer Films by Means of the Spin Pumping. Here we Investigate the ISHE Induced by the Spin Pumping on Pt/Ti/Y 3 Fe 5 O 12 and Pt/Ti/BiY 2 Fe 5 O 12 Films, where the Y 3 Fe 5 O 12 (BiY 2 Fe 5 O 12 ) and Pt Layers Are Separated by Thin Ti Layer. In these Systems, we Measured Electric Voltage due to the ISHE in Pt/Ti/Y 3 Fe 5 O 12 (BiY 2 Fe 5 O 12 ) Systems at Room Temperature. The Experimental Results Show that the ISHE Signal Disappears in the Pt/Ti/Y 3 Fe 5 O 12 (BiY 2 Fe 5 O 12 ) Systems. These Results Indicate that the Spin Pumping between Y 3 Fe 5 O 12 (BiY 2 Fe 5 O 12 ) and Ti Layer Is Suppressed. This Method Enables us to Suppress the Spin Currents without Non-Magnetic Insulators.

Resin-embedded sections and paired block surface of corn starch granules were observed using atomic force microscopy (AFM) and scanning electron microscopy to analyze the fine inner structure of starch granules and observe artifacts. Wrinkles were formed on the starch surfaces because of shear stress caused by the knife. Sectioned starches were isotropically expanded by water, and the growth rings and cracks between the growth rings were observed only on the sections. From this result, it was considered that the growth rings clearly showed shrinkage and/or corrosion of both edges of the ring during drying of the sections. Moreover, many small particles (width, ∼30 nm; height, several nanometers) were clearly observed on the growth rings. These particles could be single clusters (∼10 nm) of amylopectin molecules, considering the effect of the AFM tip radius.

Fluorescence banding has been used to classify chromosomes, except those of barley. Four of the seven barley chromosomes are indistinguishable by length or arm ratio. C-banding has been used for classification; however, it requires a long aging period. Here, we describe a new fluorescence banding method for barley. The chromosomes are treated with warm acetate followed by staining with a fluorescent dye, YOYO-1. Using this method, all seven barley chromosomes can be clearly distinguished. Atomic force microscopy and scanning near-field microscopy analyses revealed that the surfaces of the banded chromosomes were flat, indicating that the fluorescence intensity reflected the internal DNA density or condensation of chromatin.

We applied a novel silanized mica substrate with an extremely flat surface constructed according to Sasou et al. (Langmuir 19, 9845-9849 (2003)) to high-resolution detection of a specific gene on a DNA fiber by scanning near-field optical/atomic force microscopy (SNOM/AFM). The interaction between the substrate and fluorescence-dye conjugated peptide nucleic acid (PNA) probes, which causes fluorescence noise signal, was minimal. By using the substrate, we successfully obtained a fluorescence in situ hybridization signal from the ea47 gene on a λphage DNA labeled with an Alexa 532-conjugated 15-base PNA probe. As the results, no fluorescence noises were observed, indicating that the surface adsorbed almost none of the PNA probe. The combination of the substrate and SNOM/AFM is an effective tool for visualizing DNA sequences at nanometer-scale resolution.

Verrucous carcinoma は高分化型有棘細胞癌の一亜型で，緩徐に発育することを特徴とする比較的稀な疾患である。局所への浸潤傾向を示すが，リンパ節転移や遠隔転移は少ない。全体の構築が分かるように組織を採取しないと，診断が困難なことも多い。今回我々は84歳男性の右第4趾間に生じたverrucous carcinomaを経験したので報告する。数年間，良性の皮膚病変として冷凍凝固による加療を受けていたが，再発を繰り返すため，当院を受診した。当院初診時の皮膚生検でも悪性所見がなかったが，臨床像からは悪性を否定できなかったため全摘術を行った。全摘標本の病理組織学的所見よりverrucous carcinomaと診断し，拡大切除を施行，第4，5趾は切断した。術後15ヵ月を経た現在，再発は認めていない。

Methyltrimethoxysilane (MTMS)-coupled mica substrate is reportedly suitable for fixing and straightening of DNA, but 3-aminopropyltriethoxysilane (APTES)-coupled mica substrate has been found less suitable. On MTMS-coupled mica substrate, the straightness of fixed DNA was sufficient, and the adsorption of contaminants was not observed using fluorescence microscopy and atomic force microscopy. For the APTES-coupled mica substrate, however, aggregated or curved DNA and adsorption of contaminants were observed. To clarify the surface factors that are responsible for this suitability, we analyzed the surface free energies of these substrates using the extended Fowkes theory. In each of the surface free energy components, the dispersion force component in the MTMS-coupled mica substrate was lower than that in the APTES-coupled mica substrate. The ratio of the polar force component on the MTMS-coupled mica substrate was about one order of magnitude on the APTES-coupled mica substrate. In addition, the ratio of the hydrogen-bonding force component for the MTMS-coupled mica substrate was about two times larger than that of the APTES-coupled mica substrate. These results suggest that the polar force and hydrogen-bonding force components are important factors for the fixation and straightening of DNA and that the dispersion force components influence the production and adsorption of contaminants.

In starch granule, although the molecular structures of amylose and amylopectin are almost becoming clear, their hierarchic structures in molecular level is stillunknown. Recently, a blocklet model that the starch granule consists of two kinds of blocklets with different size (∼500 nm and ∼50 nm) and crystallinity (hard and soft) was proposed, and the both sizes of blocklets have been observed by scanning electron microscopy (SEM). However, by atomic force microscopy (AFM), only the smaller size (∼50 nm) of blocklet has been detected. Thus, we developed a resin-embedded section method specific to starch granule observation. As a result, we successfully observed both the big blocklet and small blocklet were clearly observed in corn starch granules by AFM. Furthermore, we found a new structure with fibrous form (10∼12 nm in height) at a center of a granule, which might be a cluster structure of amylopectin molecule with a diameter of 10∼15 nm.

The soft-nanotechnology that treats softmatter in nanometer scale needs to develop proper and original techniques for analyzing and handling the softmatter. The surface analysis of solids under vacuum condition has made remarkable progress. However, softmatter samples in vacuum atmosphere change their original properties, and are easily damaged by high energy probe beams. Thus, it is not suitable to observe “raw state” of the softmatter samples by the conventional surface analysis techniques. Scanning probe microscopy (SPM) such as atomic force microscopy (AFM) can analyze softmatter samples in moderate condition in air and water. Optical probe technique such as infrared absorption spectroscopy (IRAS) is also nondestructive method to analyze the softmatter samples. Scanning near-filed optical microscopy (SNOM), combined SPM and optical spectroscopy, enables the simultaneous measurements of topography and optical information. Here, we briefly introduce outlines of AFM, MIR-IRAS (Multiple internal reflection IRAS), and SNOM and examples of their applications.

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The predominant method for DNA cloning is by propagation in biological hosts, but this method has limitations because certain sequences are difficult to clone using any combination of available hosts or vectors. Recently, multiply-primed rolling circle amplification (MPRCA) has been applied to overcome the problems of the DNA cloning via host cells. However, when MPRCA is used to amplify from minute quantities of DNA template, the products are mostly by-product DNA molecules generated by false priming and primer dimer formation. This study demonstrates that MPRCA using random RNA primers—instead of DNA primers—blocked the synthesis of by-products and succeeded in amplifying one copy of a circular DNA molecule more than 1012-fold to give microgram quantities of amplification product without using submicroliter reaction volumes. Furthermore, a ligation strategy was elaborated to circularize only the desired DNA sequence and eliminate undesired ligation-products. A combination of these methods was able to amplify and ligate a large construct without undesired DNA sequences and at microgram quantities within one day. Therefore, these methods have the possibility to improve DNA cloning techniques that have been restricted by the limitations of PCR methods or by the host cell.

Recently, in the field of genome sequencing, next generation DNA sequencers are becoming available, and even the genome sequence analysis of a higher animal and/or a plant can be finished within a short period. However, the next generation DNA sequencers can only read relatively short sequence, which causes difficulty in the assemble step (i.e. reconstruction of original genomic sequence) and makes it labor and time consuming process and results in the long time requiring for gene isolation and breeding. When a target organism does not have enough genomic information, the assemble becomes much harder or not possible. This difficulty is arisen from the absence of positional index of the obtained nucleotide sequences by a whole genome shot-gun method. Therefore, we started the development of a new, scanning probe microscope based genome analysis method that can acquire the nucleotide sequences with positional indexes on the genome.

We developed a method to measure the rupture forces between antibody and antigen by atomic force microscopy (AFM). Previous studies have reported that in the measurement of antibody-antigen interaction using AFM, the specific intermolecular forces are often obscured by nonspecific adhesive binding forces between antibody immobilized cantilever and substrate surfaces on which antigen or nonantigen are fixed. Here, we examined whether detergent and nonreactive protein, which have been widely used to reduce nonspecific background signals in ordinary immunoassay and immunoblotting, could reduce the nonspecific forces in the AFM measurement. The results showed that, in the presence of both nonreactive protein and detergent, the rupture forces between anti-ferritin antibodies immobilized on a tip of cantilever and ferritin (antigen) on the substrate could be successfully measured, distinguishing from nonspecific adhesive forces. In addition, we found that approach/retraction velocity of the AFM cantilever was also important in the reduction of nonspecific adhesion. These insights will contribute to the detection of specific molecules at nanometer scale region and the investigation of intermolecular interaction by the use of AFM.