The functional identification of individual genes and/or cells in multicellular organism will give us invaluable pharmacological information for translational research into drug target discovery. Silencing of specific gene expression by RNA interference (RNAi) has now become a widely used tool for assessing the gene function in a fast and easy manner. An important advance in the RNAi field was the discovery that plasmid vector-mediated expression of short hairpin RNA (shRNA) can substitute for synthetic small interference RNA (siRNA) s in vitro and in vivo. But the constitutive and/or ubiquitous knock-down of target gene by this method have still limited the utility, especially if the inhibition of target gene leads to lethality, which prevents in vivo functional analysis. To overcome these limitations, the time (or period)- and cell (or tissue)- specific regulation of shRNA expression should have been required as the double-conditional mice. Conventional protocols for such an artificial regulation of gene expression in vivo are based on the tetracycline (Tet)-controlled reversible system for the time- and the Cre-loxP system for cell-specificities respectively, and the double-conditional transgenic mice are expected by mating their two lines of genetic engineered mice at last. Therefore, we have designed the "

-in " type single double conditional shRNA expression vectors having both systems within a single vector format to make the transgenic mice with double conditional RNAi without much effort. In these vectors, we have also tried to avoid the potential toxic effects of Cre not only by the use of self-deleting Cre, which deletes itself from the host cell after it is expressed, but also by the modifications of Cre recombinase as Cre-MODC, DD-Cre or Cre-ERT2. We have also applied the Piggyback (PB)-transposon system for integrating multiple functional units contained the core of these single double conditional shRNA expression vectors into genomes with the use of PB transposase mRNA which was short-lived without integration of PB transposase gene itself. Such the low toxic and highly integrated systems may be useful tools for making spatio-temporal controlled transgenic mice in the pronuclear injection steps of these transgenes into the fertilized eggs.

抄録全体を表示

An

-in- unpowered Mars rover functions as an entry, descent, and landing (EDL) system during atmospheric entry, an airbag at landing, and an unpowered rover at Mars surface with an inflated spherical structure. This rover can access attractive but risky areas, where typical rovers cannot access. This study considers the feasibility of this rover in terms of EDL and roving, and demonstrates the required rover configuration that enables both robustness in EDL and mobility on Mars surface.

抄録全体を表示

The purpose of this study was to evaluate the effect of air-blowing duration on the bonding performance of -in- systems using the same pressure (0.25 MPa). Three -in- systems were: EB (Easy Bond, 3M ESPE, USA), BB (BeautiBond, Shofu Inc., Japan) and GBp (G-Bond plus, GC Corporation, Japan). After adhesive application, the 3 systems were air-blown thereafter using 7 different durations (5 s, 10 s, 15 s, 20 s, 25 s, 30 s and 35 s). Bond strengths to dentin were determined using µTBS test after 24 h water storage. In addition, evaluation of both the resin-dentin interface and the fractured surface on the dentin side were performed by SEM. The maximum µTBS for each system, BB (40.4±14.8 MPa), EB (79.8±16.5 MPa), and GBp (47.3±17.6 MPa), were recorded with 15 s, 15 s and 25 s air-blowing duration respectively. Under the same air-pressure, the air-blowing duration could affect evaporation and the thickness of the adhesive layer, which contributed to the different bond strengths.

抄録全体を表示

【要旨】心臓手術の術式変更,緊急手術を行う場合,技士が短時間で人工心肺回路を組み立て灌流補助することは,患者の生命を左右する重要なことである。しかし,従来の回路は,準備を行い灌流開始するまで時間を要してしまい,回路組み立て時間の短縮化が望まれていた。そこで今回,回路の機能的な部分は変更せずに,メーカー側で事前に接続されたPre connect回路と,心筋保護液回路を含めた回路セットを1個のトレイの中に組み込んだ in System回路を新たに製作した。その結果,回路の準備から充填までの時間短縮が可能となり,仕事の効率化・合理化が行われ緊急手術などに活用できる回路が完成した。このことが実現となった背景として,メーカーの人工心肺関連商品の充実化とリザーバー付き人工肺の国内滅菌が可能となったことがあげられる。

抄録全体を表示

近年,接着性レジンシステムのエッチング,プライミング,ボンディングの操作を簡便に一度に行える-in- adhesiveの研究開発が盛んである。これらの材料を小窩裂溝填塞材(シーラント)として使用する可能性を検討するため研究を行った。その結果,-in- adhesiveのエナメル質に対する接着力はシーラント材の接着力と有意差はなく,齲蝕予防効果に必要なフッ素の徐放と取り込みの機能が付与されていた。これらの材料は酸性モノマーを含有するため,細胞毒性試験では未重合モノマーの細胞培養液中への溶出によりpHの低下を示し,細胞毒性がわずかに発現した。xeno CF II Bond と AQ Bond plusの細胞毒性が強かった。-in- adhesiveは十分なエナメル質接着力を示し,小窩裂溝に侵入し,操作が簡便であるためシーラント材として応用が可能である。

抄録全体を表示

It has been reported that the adhesive resin layer of -in- adhesives is permeable and allows the formation of a water channel or water tree. The effects of a smear layer on the ultra-structure of bonding interface created with three -in- adhesives and a resin composite were evaluated in this study. Dentin surface was ground with of 180-, 600-, or 2000-grit SiC paper to produce different smear layer thickness. The bonding interfaces were observed under a transmission electron microscope without demineralization. Voids of various sizes and water channels were found at the bottom of the adhesive resin layer along the bonding interface of SiC 180-grit dentin, while SiC 2000-grit dentin did not produce any voids. The results showed that the voids were possibly related to water that had penetrated from the underlying dentin. A smear layer may have an adverse influence on the bonding performance of -in- adhesives to dentin.

抄録全体を表示

Photoelectron spectroscopy is our main tool to explore the electronic structure of novel material systems, the properties of which are often determined by an intricate interplay of competing interactions. Elucidating the role of this interactions requires studies over an extensive range of energy, momentum, length, and time scales. We show that immersion lens-based momentum microscopy with spin-resolution is able to combine these seemingly divergent requirements in a unifying experimental approach. We will discuss applications to different areas in information research, for example, resistive switching and spintronics. The analysis of resistive switching phenomena in oxides requires high lateral resolution and chemical selectivity, as the processes involve local redox processes and oxygen vacancy migration. In spintronics topological phenomena are currently a hot topic, which lead to complex band structures and spin textures in reciprocal space. Spin-resolved momentum microscopy is uniquely suited to address these aspects.

抄録全体を表示

レジン接着システムの良好な歯質接着は, エッチング, プライミング, ボンディングという基本的な3歯面処理を経て獲得されている。近年では, 接着メカニズムの探求や応用材料の開発改良が進み, 基本的歯面処理の前2者または後2者を合わせて2ステップとしたシステムが臨床において主流となっている。さらに最近では, 処理の1ステップ化や操作性の向上を図りながら優れた歯質接着性の獲得を求めた-in- adhesive systemが登場しはじめている。そこで本研究では, これら-in- adhesive systemの歯頸部歯質に対する接着性を明らかにすることを目的に, 規格化V字状窩洞における辺縁封鎖性とエナメル質および象牙質に対する狭小面接着強さの点から検討を行った。
したがって-in- adhesive systemの歯頸部歯質に対する接着性は対照としたSBに比べ, 処置直後の接着強さ獲得能の点においては同等もしくは優れていると考えられるが, 過酷な口腔内環境を想定した複合負荷条件下においては歯面や処理材の成分および物性などの違いによって辺縁封鎖性が影響を受けるものと考えられる。

抄録全体を表示

Gene silencing by RNA interference （RNAi） is widely used for assessing gene function. An important advance in the RNAi field was the discovery that plasmid-based RNAi can substitute for synthetic small interfering RNA in vitro and in vivo. However, constitutive and ubiquitous knockdown of gene expression by RNAi in mice can limit the scope of experiments because this process can lead to embryonic lethality, or result in compensatory overexpression of other genes such that no phenotypic abnormalities occur. Either way, analyses of the physiological roles of the gene of interest in adult mice are not possible. To overcome these limitations, we previously constructed a double-conditional short-hairpin RNA （shRNA） expression vector that can regulate shRNA expression in a spatio-temporal manner with a tetracycline-inducible floxed stuffer sequence selectively excised by application of Cre recombinase. In this study, we aimed to modify this vector to create an -in- vector that produces double-conditional transgenic mice through a single round of gene transfer to fertilized eggs. We added a coding region for nuclear localizing Cre （NCre） recombinase with a multi-cloning site for a cell-specific promoter into the double-conditional short-hairpin RNA （shRNA） expression vector that we previously constructed. Using Escherichia coli, we confirmed successful construction of the vector. First, we confirmed isopropyl-β-D-thiogalactopyranoside-induced expression of NCre recombinase through the lac operon as a specific promoter by western blotting. Second, we confirmed functional recombination of the floxed sequence of loxP-like TATA-lox by analysing restriction enzyme-digested fragments. This -in- double-conditional shRNA expression vector will be useful for reversible in vitro and in vivo knockdown of target gene expression, in target cells via promoter-specific expression of NCre, and at specific times by tetracycline application.

抄録全体を表示

The outcome of patients with severe traumatic brain injury presenting with dilated pupils is generally poor, and decompression surgery at the earliest possible is warranted in such cases. Our strategy for managing patients with severe traumatic brain injury comprises completing the series of treatment provided at prehospital, trauma bay, intensive care unit, and rehabilitation center at a single department in a hospital (

–in– Strategy). Herein, we report the case of a patient with severe traumatic brain injury who was successfully treated using this strategy and who survived to hospital discharge with a good neurological outcome. A 19–year–old male was injured in a traffic accident at midnight, following which a physician–staffed ambulance was dispatched from our hospital. When the physician contacted the patient at the emergency site, the patient was comatose and had developed Cushing reflex. The physician notified the trauma bay for emergency surgery at the emergency site.

Soon after arriving at the trauma bay, the patient underwent head computed tomography, revealing bilateral acute subdural hematoma with multiple contusions. An emergency trepana­tion was performed 58 min after the patient’s arrival at the hospital because his pupils were dilated. Subsequently, the patient was transferred to the operation room and decompressive craniotomy performed 74 min after his arrival. The patient recovered to eat and walk on his own and survived to discharge at day 84 with a Glasgow Outcome Scale category of moderate disability.

抄録全体を表示

Objectives: Raman spectroscopy yields precise information, not only regarding the secondary structure of proteins but also regarding the discrimination between normal and malignant tissues. There is, however, no standard measurement method. We evaluated the use of a miniaturized, handheld,

-in- Raman spectrometer with a 1064-nm laser excitation source for the diagnosis of colorectal cancer. The ultimate goal is real-time, in vivo diagnosis. Methods: Tissue samples were obtained from 20 patients who underwent surgery for colorectal cancer. The samples were irradiated with the portable Progeny™ Raman spectrometer, with which the Raman spectra were also obtained. We searched for characteristic Raman shifts and examined whether these shifts could distinguish the cancer tissues. To improve accuracy, we divided the spectra into 100 cm⁻¹ bands and applied principal component analysis (PCA) to each range. We evaluated the contribution of each range for cancer discrimination. Results: Intensities at 1261 and 1427 cm⁻¹ differed significantly between the normal tissues and cancer tissues, but these did not efficiently discriminate the cancer tissues. However, we were able to identify the characteristic spectral range in fingerprint regions; accuracy was 85.1%. Conclusions: Use of the -in- type Raman spectrometer can efficiently discriminate colorectal cancer, not on the basis of the intensities at 1261 and 1427 cm⁻¹ but rather on the basis of PCA. Thus, Raman spectroscopy performed using a handheld device has potential to become a clinically powerful tool for producing high-quality data, obtaining highly reproducible measurements, and thus accurately diagnosing colorectal cancer.

抄録全体を表示

The self-etching primer adhesives examined in the present study were aqueous mixtures of 5wt% MDP, 35wt% HEMA, 3wt% 2-hydroxyl-3-(3, 4-dimethyl-9-oxo-9H-thioxanthen-2-yloxy)-N, N, N-trimethyl-1-propanaminium chloride (QTX), 0.5wt% ethyl-p-(N, N-dimethylamino) benzoate (EDM), and 30wt% water-soluble dimethacrylate (i. e., 2, 2-bis (4-methacryloxy polyethoxypheny) propane (BPE), Glycerol dimethacrylate (GMR), or Glycerol methacrylate acrylate (GAM)). Self-etching primer adhesive was directly applied to polished bovine teeth. Tensile bond strength of composite resin to bovine teeth was measured after one-day immersion in water at 37°C. Differences in water-soluble dimethacrylate monomer did not produce any significant differences in tensile bond strength to dentin or enamel. The use of N-phenyliminodiacetic acid (PIDAA) instead of EDM showed significantly higher bond strength to dentin and enamel. In conclusion, GMR and GAM were useful as water-soluble dimethacrylates while the combination of PIDAA and QTX was effective in improving the bond strength of composite resin to teeth in an -in- bonding system.

抄録全体を表示

In projection mapping on a three-dimensional object with a free-form curved surface, it is necessary to perform geometric correction of the projected video image pixel by pixel. The adjustment of the video image, including such correction, is extremely complicated and time-consuming, and it is a major burden on the creator. We have developed an application that can set up projection mapping in a short time with almost only mouse operations by integrating these series of tasks. This application enables accurate mapping of three-dimensional objects with free-form surfaces using an inexpensive projector and camera, without the use of special equipment. It can also be used in conjunction with commercially available video content creation applications.

抄録全体を表示

This study was evaluated the tensile bond strength (TBS) of resin cements to bovine dentin resin-coated with -in- adhesive systems. Each of the dual-polymerizing resin cements; Link Max, Clearfil Esthetic Cement, Bistite II and Chemiace II were used to bond indirect resin disks to bovine dentin, as control, or coated by single-application or by dual-application of an adhesive system from the same manufacturer; G-Bond, Clearfil Tri-S Bond, Tokuyama Bond Force and Hybrid-Coat (n=10). After 24-hour water storage, TBSs were measured. The fracture pattern and the adhesive interface were observed using an SEM. Dual-application of the adhesive yielded significantly higher TBSs compared to control and single-application groups for all materials (p<0.001). From the limited information of this study, it was concluded that dual-application of -in- adhesive systems created a thin coating on dentin, and significantly improved the bond strengths of resin cements.

抄録全体を表示

This study evaluated the effect of resin coating using an -in- adhesive system on the dentin bond strength of a dual-cure resin cement after different curing modes. Human molars were ground to obtain flat dentin surfaces and divided into three groups: untreated as a control and resin-coated with either a single- or double-application of an -in- adhesive (Tokuyama Bond Force). The specimens were bonded to indirect composite disks using a dual-cure resin cement (Bistite II) activated by dual-cure or self-cure modes. Each specimen was sectioned into beams for the microtensile bond strength test. The data were analyzed by two-way ANOVA with Bonferroni’s correction (p=0.05). Resin coating with a double-application of the -in- adhesive system significantly improved the bond strength of the dual-cure resin cement to dentin. In addition, dual-curing of the resin cement enhanced the bond strengths to dentin.

抄録全体を表示

This study evaluated the capability of a novel fluoride-releasing, -in- adhesive system on forming inhibition layer (radio-opaque layer) as compared with other adhesive systems. Dentin surface was treated with Imperva bond (TB), FL-BOND (FB), Reactmer bond (RE), or FL-BOND S-1 (FS) (which is a novel system). Untreated specimens were categorized as nonbonding group (NB). After storing for 10 days in de-ionized water, the specimens were cut into halves perpendicularly to the pulp chamber and immersed in a buffered demineralizing solution for four days. Longitudinal sections were cut and microradiographed. The width of inhibition layers adjacent to the adhesive surface—at a depth of 50μm under the demineralization surface—was analyzed. Microradiography revealed distinct inhibition layers adjacent to the experimental surfaces of FB, RE, and FS. No inhibition layers were observed in NB and TB. In particular, the width of the inhibition layer of FS (12.5μcm) was significantly greater than those of FB and RE. These results indicated that a newly developed -in- adhesive system, FS, may have a superior ability of forming inhibition layers adjacent to cavity walls, and that it may also protect dentin against further demineralization in case of secondary marginal caries.

抄録全体を表示

-in- self-etching model adhesives, HEMA-free and without phase separation were designed. Enamel and dentin bond strength of compounds with 4-META or 4-MET (28%), dissolved in acetone and water (5-8%) were > 20 MPa. Marginal dentin cavity adapation was almost perfect.

抄録全体を表示

新規オールインワンアドヒーシブシステムＴｒｉ_-_Ｓ　Ｂｏｎｄの象牙質接着界面を透過型電子顕微鏡にて観察した。また脱灰試料と未脱灰試料を用いることにより、接着界面の超微細 構造を解析した。

抄録全体を表示