Transient receptor potential ankyrin 1 agonists improve intestinal transit in a murine model of postoperative ileus.

BACKGROUND: Stimulation of transient receptor potential ankyrin 1 (TRPA1), which abundantly expressed in enterochromaffin cells (ECC), has been reported to exert apparently contradictory results in in vitro contractility and in vivo gastrointestinal (GI) transit evaluations. The pharmaceutical-grade Japanese traditional medicine daikenchuto (TU-100) has been reported to be beneficial for postoperative ileus (POI) and accelerate GI transit in animals and humans. TU-100 was recently shown to increase intestinal blood flow via stimulation of TRPA1 in the epithelial cells of the small intestine (SI). METHODS: The effects of various TRPA1 agonists on motility were examined in a manipulation-induced murine POI model, in vitro culture of SI segments and an ECC model cell line, RIN-14B. KEY RESULTS: Orally administered TRPA1 agonists, aryl isothiocyanate (AITC) and cinnamaldehyde (CA), TU-100 ingredients, [6]-shogaol (6S) and γ-sanshool (GS), improved SI transit in a POI model. The effects of AITC, 6S and GS but not CA were abrogated in TRPA1-deficient mice. SI segments show periodic peristaltic motor activity whose periodicity disappeared in TRPA1-deficient mice. TU-100 augmented the motility. AITC, CA and 6S increased 5-HT release from isolated SI segments and the effects of all these compounds except for CA were lost in TRPA1-deficient mice. 6S and GS induced a release of 5-HT from RIN-14B cells in a dose- and TRPA1-dependent manner. CONCLUSIONS & INFERENCES: Intraluminal TRPA1 stimulation is a potential therapeutic strategy for GI motility disorders. Further investigation is required to determine whether 5-HT and/or ECC are involved in the effect of TRPA1 on motility.

Investigation of the influence of coadsorbent dye upon the interfacial structure of dye-sensitized solar cells.

The interface between Ru(tcterpy)(NCS)3TBA2 [black dye (BD); tcterpy = 4,4',4″-tricarboxy-2,2':6',2″-terpyridine, NCS = thiocyanato, TBA = tetrabutylammonium cation] and nanocrystalline TiO2, as found in dye-sensitized solar cells, is investigated by soft-X-ray synchrotron radiation and compared with the adsorption structure of cis-Ru(Hdcbpy)2(NCS)2TBA2 (N719; dcbpy = 4,4'-dicarboxy-2,2'-bipyridine) on TiO2 to elucidate the relationship between the adsorption mode of BD and the photocurrent with and without coadsorbed indoline dye D131. The depth profile is characterized with X-ray photoelectron spectroscopy and S K-edge X-ray absorption fine structure using synchrotron radiation. Both datasets indicate that one of the isothiocyanate groups of BD interacts with TiO2 via its S atom when the dye is adsorbed from a single-component solution. In contrast, the interaction is slightly suppressed when D131 is coadsorbed, indicated by the fact that the presence of D131 changes the adsorption mode of BD. Based upon these results, the number of BD dye molecules interacting with the substrate is shown to decrease by 10% when D131 is coadsorbed, and the dissociation is shown to be related to the short-circuit photocurrent in the 600-800 nm region. The design of a procedure to promote the preferential adsorption of D131 therefore leads to an improvement of the short-circuit current and conversion efficiency.

Gate control of electronic phases in a quarter-filled manganite.

Electron correlation often produces a variety of electrically insulating states caused by self-organization of electrons, which are particularly stable at commensurate fillings. Although collapsing such ordered states by minute external stimuli has been a key strategy toward device applications, it is difficult to access their true electronic phase boundaries due to the necessity of fine-tuning of material parameters. Here, we demonstrate the ambipolar resistance switching in Pr(1-x)Sr(x)MnO3 thin films (x = 0.5; an effectively 1/4-filled state) by quasi-continuous control of the doping level x and band-width W using gate-voltage and magnetic field, enabled by the extreme electric-field formed at the nanoscale interface generated in an electrolyte-gated transistor. An electroresistance peak with unprecedented steepness emerges on approaching a critical point in the x-W phase diagram. The technique opens a new route to Mott-insulator based transistors and to discovering singularities hitherto unnoticed in conventional bulk studies of strongly correlated electron systems.

Case of progressive facial hemiatrophy with cervical sympathetic hyperactivity as underlying aetiology.

OBJECTIVE: We report a case of progressive facial hemiatrophy with cervical sympathetic hyperactivity as a possible underlying aetiology, based on clinical findings, three-dimensional computed tomography and thermographic imaging. METHODS: We present a case report in which we describe the investigation and clinical course of progressive facial hemiatrophy, and we also review the world literature on this condition. RESULTS: To our knowledge, this is the first report in the world literature of progressive facial hemiatrophy with cervical sympathetic hyperactivity indicated as a possible underlying aetiology, based on clinical findings, three-dimensional computed tomography and thermographic imaging. CONCLUSION: This syndrome may lead to atrophy of the subcutaneous adipose tissue with hyperfunction of the vegetative system. Although this is a rare syndrome, otolaryngologists should be aware of its symptoms, aetiology and treatment.

Photoinduced metal-to-insulator transition in a manganite thin film.

A persistent photoinduced metal-to-insulator transition has been confirmed in a manganite thin film, Pr_(0.55)(Ca_(0.75)Sr_(0.25))_(0.45)MnO3, near a multicritical point by monitoring with transport measurements and x-ray photoemission spectroscopy. Together with the previously reported reverse effect, the photoinduced insulator-to-metal transition, it is found that the relative stability of the metallic and insulating phases interchanges around 80 K in the middle of a very wide hysteresis loop, which is a manifestation of the large potential barrier due to the long-range elastic energy. It is shown that photons are much more effective in overcoming the barrier via the electronically excited intermediate states than via the heat mode.

Intrinsic colossal magnetoresistance effect in thin-film Pr0.5Sr0.5MnO3 through dimensionality switching.

A homogeneous colossal magnetoresistance (CMR) effect at low temperatures has been found in a thin-film perovskite manganite Pr0.5Sr0.5MnO3. The transition is driven not by the spin alignment as in usual CMR in bulk samples but by the localization-delocalization transition switched by the change in the effective dimensionality. Two-dimensional (x2-y2)-orbital ordering enhanced by the substrate strain is essential for the stabilization of the insulating localized state, which is on the verge of the first-order transition to the three-dimensional metallic ferromangetic state.

Novel orbital ordering induced by anisotropic stress in a manganite thin film.

A novel structure of orbital ordering is found in a Nd0.5Sr0.5MnO3 thin film, which exhibits a clear first-order transition, by synchrotron x-ray diffraction measurements. Lattice parameters vary drastically at the metal-insulator transition at 170 K (= T(MI)), and superlattice reflections appear below 140 K (= T(CO)). The electronic structure between T(MI) and T(CO) is identified as A-type antiferromagnetic with a d(x2-y2) ferro-orbital ordering. The new type of antiferro-orbital ordering characterized by the wave vector (1/4 1/4 1/2) in cubic notation emerges below T(CO). The accommodation of the large lattice distortion at the first-order phase transition and the appearance of the novel orbital ordering are brought about by the anisotropy in the substrate, a new parameter for the phase control.

Persistent and reversible all-optical phase control in a manganite thin film.

Persistent and reversible optical phase control has been achieved in a manganite thin film through a careful choice of the composition of Pr1-x(Ca1-ySr(y))xMnO3 near a multicritical point. Pulsed laser light brings the lower temperature metallic phase out of the higher temperature charge-ordered insulator, while a cw light reverses the effect by heating. We clearly demonstrate the two competing roles played by light, heating, and excitation across the charge gap, which are important in both the application and the understanding of the physics of electron correlation.

Photocontrol of dynamic phase transition in the charge-density wave material K(0.3)MoO(3).

A photoinduced effect has been studied in the quasi-one-dimensional conductor K(0.3)MoO(3). A weak photoexcitation of the sample can drastically affect the dynamic phase transition of the charge-density wave (CDW) motion from slide to creep. The origin of the photoeffect is attributed to a local destruction of the CDW which leads to the photoinduced phase slip and the redistribution of the CDW phase.

A fusion protein between rac and p67phox (1-210) reconstitutes NADPH oxidase with higher activity and stability than the individual components.

Activation of the phagocyte NADPH oxidase, a superoxide-generating enzyme, involves assembly of cytosolic p47(phox), p67(phox), and rac with the membrane-associated cytochrome b(558). Following cell-free activation, enzymatic activity is highly labile [Tamura, M., Takeshita, M., Curnutte, J. T., Uhlinger, D. J., and Lambeth, J. D. (1992) J. Biol. Chem. 267, 7529-7538]. In an attempt to stabilize the activity and to investigate the nature of the complex, we have produced fusion proteins between rac and a C-terminal truncated form of p67(phox) (residues 1-210, 67N), which is a minimal active fragment. In a cell-free system, a fusion protein 67N-rac had higher activity and a 3-fold higher affinity than the individual cytosolic proteins, and 67N-Ser3-rac, which has a longer linker, showed a similar activity with the individual proteins. In contrast, rac-67N, a fusion in the opposite orientation, showed considerably lower activity. The enzyme activity reconstituted with 67N-rac showed a 10-fold higher stability and a lower K(m) for NADPH than the individual components. In the absence of p47, 67N-rac fusion protein at a high concentration showed nearly full activation, which was higher than that with the individual components. These results indicate that covalent binding between p67N and rac in the correct order produces a more stable complex than the individual components, suggesting that interactions among the subunits significantly influence the duration of the oxidase activation. On the basis of these findings, we propose a model for the topology among rac, 67N, and cytochrome b(558).

Expression and characterization of recombinant human antithrombin III in Pichia pastoris.

Antithrombin III (ATIII) is a member of the serpin superfamily and a major regulator of the blood coagulation cascade. To express recombinant human ATIII (rATIII) in the methylotrophic yeast Pichia pastoris, we constructed an rATIII expression plasmid which contained the ATIII cDNA encoding mature protein region connected with the truncated mAOX2 promoter and the SUC2 secretion signal, introduced it into the P. pastoris genome, and screened for a single copy transformant. The secretion of rATIII from the transformant reached a level of 320 IU/L in the culture broth at 169 h. From the culture-supernatant, rATIII was purified to over 99% by heparin-affinity chromatography and other column chromatography methods. We characterized rATIII and compared it with human plasma-derived ATIII (pATIII). The purified rATIII possessed correct N-terminal amino acid sequence, and its molecular weight by SDS-PAGE of 56,000 Da was slightly different from the 58,000 Da of pATIII. Sequence and mass spectrometry analysis of BrCN fragments revealed that posttranslational modifications had occurred in rATIII. O-linked mannosylation was found at Ser 3 and Thr 9, and in some rATIII molecules, modification with O-linked mannosyl-mannose had probably occurred at Thr 386, close to the reactive center. Although the heparin-binding affinity of rATIII was 10-fold higher than that of pATIII, its inhibitory activity against thrombin was only half. As the conformation of rATIII and pATIII by circular dichroism spectroscopy was similar, O-glycosylation in the reactive center loop was assumed to be mainly responsible for the decreased inhibitory activity. pATIII can inactivate thrombin through formation of a stable thrombin-ATIII complex, but rATIII modified with O-glycosylation in the reactive center loop may act as a substrate rather than an inhibitor of thrombin.

Nonlinear magneto-optical properties of colossal magnetoresistive manganites.

Pr(1--x)CaxMnO(3) and Nd(1--x)SrxMnO(3) were investigated with three-photon difference frequency generation (DFG). This method allows one to determine both the crystalline and the magnetic symmetry. In the highly ordered low-temperature phase of Nd(0.50)Sr(0.50)MnO(3), a DFG contribution coupling simultaneously to antiferromagnetic and charge ordering was observed and used to reveal the formation of domains. Thus, magnetically induced three-photon processes are introduced into the fields of both nonlinear magneto-optics and colossal magnetoresistance as a powerful new method.

Fused p47phox and p67phox truncations efficiently reconstitute NADPH oxidase with higher activity and stability than the individual components.

Activation of the neutrophil NADPH oxidase occurs via assembly of the cytosolic regulatory proteins p47(phox), p67(phox), and Rac with the membrane-associated flavocytochrome b(558). Following cell-free activation, enzymatic activity is highly labile (Tamura, M., Takeshita, M., Curnutte, J. T., Uhlinger, D. J., and Lambeth, J. D. (1992) J. Biol. Chem. 267, 7529-7538). To try to stabilize the activity and investigate the nature of the complex, fusion proteins between p47N-(1-286) and p67N-(1-210) were constructed. In a cell-free system, a fusion protein, p67N-p47N, had an 8-fold higher efficiency and produced a higher activity than the individual proteins, and also resulted in an 8-fold improved efficiency for Rac and a lowered K(m) for NADPH. O(2) generating activity was remarkably stabilized by using p67N-p47N. The cytosolic proteins fused in the opposite orientation, p47N-p67N, showed similar activity and stability as individual proteins, but with a 4-fold improved efficiency compared with the individual cytosolic factors. In the system efficiency for Rac and affinity for NADPH were also higher than those with the nonfused components. Interestingly, the p67N-p47N showed nearly full activation in the absence of an anionic amphifile in a cell-free system containing cytochrome b(558) relipidated with phosphatidylinositol- or phosphatidylserine-enriched phospholipid mixtures. From the results we consider multiple roles of anionic amphifiles in a cell-free activation, which could be substituted by our system. The fact that a fusion produces a more stable complex indicates that interactions among components determine the longevity of the complex. Based on the findings we propose a model for the topology among p47N, p67N, and cytochrome b(558) in the active complex.

Improvement of vitamin B(12) fermentation by reducing the inhibitory metabolites by cell recycle system and a mixed culture.

The major problem in vitamin B(12) production using Propionibacterium is the growth inhibition of the cell due to the accumulation of inhibitory metabolites such as propionic acid and acetic acid. In the present paper, we considered several approaches of controlling the propionic acid concentration at low level. Namely: (1) the periodic cultivation of Propionibacterium where dissolved oxygen (DO) concentration was alternatively changed between 0 and 1ppm; (2) cell recycle system using hollow fiber module; and (3) mixed culture using Propionibacterium and Ralstonia eutropha where the latter microorganism assimilates the propionic acid produced by the former. It was found that the productivity of vitamin B(12) was the highest for the cell recycle system, while if the performance was evaluated based on the amount of vitamin B(12) produced per medium used, the mixed culture system gave the far highest value.

Microsphere resonators strongly coupled to a plane dielectric substrate: coupling via the optical near field.

A model is proposed that describes the essential optical process in the recently observed resonant light scattering from a microsphere resonator that is strongly coupled to the substrate. The experimentally observed field patterns across the resonance can be reproduced quite well by a numerical calculation taking into account only a few vector spherical waves that are converted from nonpropagating to propagating waves at the substrate surface. Explicit consideration of the multiple-reflection effect is not necessary to reproduce the experimental results. Comparison of the experiment and the calculation suggests the splitting of degenerate resonance modes that have different azimuthal mode numbers within a single broad resonance line. These results are discussed on the basis of the strongly coupled nature of the system.

Metabolic pathway of Propionibacterium growing with oxygen: enzymes, 13C NMR analysis, and its application for vitamin B12 production with periodic fermentation.

The metabolic pathway of Propionibacterium grown under an aerobic condition is still not clear so far. In this work, cell growth, organic acid formation, vitamin B12 synthesis, and enzyme activities were determined in different aerobic cultivation systems. It was found that the propionate, which is accumulated during anaerobic cultivation, was completely decomposed when the cultivation was shifted to an aerobic condition. Moreover, pyruvate was formed in accordance with the decomposition of the propionate. Besides, more acetate was produced and a large amount of malate was formed during the aerobic cultivation. Such phenomena could be repeatedly observed in a periodic cultivation in which the dissolved oxygen concentration was alternatively controlled at 0 or 1 ppm. Enzyme analysis indicates that the regulation of organic acid formation depends on which molecule, i.e., oxygen or fumarate, serves as an electron acceptor in the respiratory chain reactions. No tricarboxylic acid cycle was found to exist in this species grown under an aerobic condition. It is evident that the randomizing pathway worked in a reversed direction in the presence of oxygen, through which the propionate is oxidized to pyruvate. The 13C NMR spectral analysis confirmed this observation.

Effect of dopamine content in rat brain striatum on anesthetic requirement: an in vivo microdialysis study.

This study investigated the changes in anesthetic requirement caused by administration of two different concentrations (0.1 microM and 1. 0 microM/h) of dopamine into the rat striatum. During the measurement of minimum alveolar concentration (MAC), each dopamine solution was continuously injected directly into rat striatum by microdialysis technique. During perfusion of the lower dose, MAC did not change. During the higher dose of dopamine, MAC decreased by approximately 30%.

Anti-HIV-1 activity of myo-inositol hexaphosphoric acid (IP6) and myo-inositol hexasulfate(IS6).

It is known that polysulfates have some anti-HIV-1 activity. We investigated the anti-HIV-1 activity of myo-inositol hexaphosphoric acid (IP6) and myo-inositol hexasulfate(IS6), low molecular weight carbohydrates. IP6 and IS6 inhibited the replication of HIV-1 in a T cell line as well as that of a freshly isolated strain in peripheral blood mononuclear cells. Neither substance inhibited HIV-1-induced giant cell formation, but addition of IS6 when infecting cells with HIV-1 inhibited the replication of HIV-1. Neither substance inhibited HIV-1 reverse transcriptase activity in vitro and no influence on late stage replication was noted. Although the mechanisms of IP6 and IS6 action remain unclear, it can be speculated that they act on HIV-1 early replicative stage. Although it is not possible to develop IP6 and IS6 themselves as anti-AIDS drugs, studies of these anti-HIV agents might be expected to provide seed for eventual production of superior drugs for AIDS treatment.

Observation of a modulation effect caused by a microsphere resonator strongly coupled to a dielectric substrate.

The effect of modulation caused by a microsphere resonator is experimentally investigated with a model system consisting of a microsphere resonator and a plane substrate. We used total internal reflection microscopy (TIRM), which is a combination of conventional optical microscopy and the total internal reflection method, and observed the intensity distribution under the resonator in the evanescent-wave incidence condition. The TIRM patterns drastically change when the wavelength of the incident beam is scanned across a resonance. The response of the system is discussed on the basis of a recent proposal of traveling-wave resonance.

Visualization of the local insulator-metal transition in Pr0.7Ca0. 3MnO3

The light-induced insulator-metal transition in the "colossal magnetoresistance" compound Pr0.7Ca0.3MnO3 is shown to generate a well-localized conducting path while the bulk of the sample remains insulating. The path can be visualized through a change of reflectivity that accompanies the phase transition. Its visibility provides a tool for gaining insight into electronic transport in materials with strong magnetic correlations. For example, a conducting path can be generated or removed at an arbitrary position just because of the presence of another path. Such manipulation may be useful in the construction of optical switches.