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To establish a new rat model of craniofacial myalgia, and to clarify which central nervous system pathways are activated in the model.Craniofacial myalgia, represented by myogenous temporomandibular disorder and tension-type headache with pericranial tenderness, is more common in female patients. The pain is thought to be a type of multifactorial disorder with several coexisting causes. To our knowledge, there are no models of craniofacial muscle hyperalgesia caused by multiple types of stimuli.We injected nerve growth factor into the trapezius muscle of female and male rats and repeatedly stimulated the masseter muscle (MM) electrically for 10 days. We determined the mechanical head-withdrawal threshold of MM and extent of phosphorylated extracellular signal-related kinase 1/2 (pERK) immunoreactivity in various regions of the lower brainstem. We conducted retrograde tract-tracing to determine the projection of mechanosensitive MM-innervating secondary neurons to the lateral parabrachial nucleus. Finally, we administered morphine in rats to determine whether increases of pERK immunoreactivity were dependent on noxious inputs.In female rats, but not male rats, the mechanical head-withdrawal threshold was decreased significantly from days 9 to 12. The number of pERK-immunoreactive neurons in the brainstem was increased significantly in female rats in the group with both stimuli compared to rats in other groups with a single stimulus. Mechanosensitive MM-innervating neurons in the brainstem projected to the parabrachial nucleus. Morphine administration blocked the increase in the number of pERK-immunoreactive neurons in both the brainstem and parabrachial nucleus.We established a model of craniofacial myalgia by combining trapezius and MM stimuli in female rats. We found mechanical hyperalgesia of the MM and activation of the pain pathway from the brainstem to parabrachial nucleus. The model reflects the characteristics of patients with craniofacial myalgia and might be helpful to clarify the pathogenic mechanisms underlying these disorders.

Background Anastomotic disorder of the reconstructed gastric conduit is a life‐threating morbidity after thoracic esophagectomy. Although there are various reasons for anastomotic disorder, the present study focused on dislocation of the gastric conduit (DGC). Methods The study cohort comprised 149 patients who underwent transthoracic esophagectomy. The relationships between DGC and peri‐ and postoperative morbidities were analyzed retrospectively. Data were analyzed to determine whether body mass index (BMI) and extension of the gastric conduit were related to DGC. Uni‐ and multivariate Cox regression analyses were performed to identify the factors associated with anastomotic disorder. Results DGC was significantly related to anastomotic leakage (P, Anastomotic disorder is a life‐threating morbidity after esophagectomy. Anastomotic leakage, stricture, and mediastinal abscess/empyema are significantly related to dislocation of the gastric conduit.

The first total synthesis of dermocanarin 2 is described. The synthesis features the construction of the anthraquinone and naphthoquinone frameworks through annulation reactions onto an axially chiral biphenyl intermediate, obtained by an enzyme-catalyzed enantioselective desymmetrization of a σ-symmetric precursor, followed by a stereoselective aldol reaction to construct the stereogenic center in the side chain.

We provide a simple model for analyzing how debt forgiveness affects the stock price of a lending bank. Our model shows that although debt forgiveness increases shareholder wealth of a bank in healthy financial condition, it decreases shareholder wealth of a bank in unhealthy financial condition. We empirically investigate the announcement effect of debt forgiveness on bank stock prices in Japanese markets. On average, lending banks experience a significant negative announcement effect with respect to debt forgiveness. Consistent with the prediction of the model, we find a negative relation between the announcement effect and the net bad loan ratio as a proxy of the unhealthiness of the financial condition of the bank.

Aim: To investigate the latest serological tendency of cytomegalovirus (CMV) in Japanese puerperal women and to estimate the sequential influence on child-bearing women in Japan. Methods: We studied 3966 puerperal women registered at a cord blood bank. Serum samples were obtained from umbilical cords at delivery and on the third day after delivery. CMV immunoglobulin G (IgG) and CMV immunoglobulin M (IgM) antibodies were determined using enzyme immunoassay kits. Scatter diagrams between seroprevalences and perinatal details were generated, and Pearson product-moment correlation coefficients (r) and coefficients of determination (R2) were calculated. Results: The overall CMV seroprevalence was 68.4% for the IgG antibody and 3.9% for the IgM antibody. IgM seroconversion had occurred separately in 0.28% of the cases without IgG seroconversion. Including outliers on scatter plots, linear decreasing trends were observed for IgG seropositivity with measurement year (r = −0.72, P

Structural changes in bacteriorhodopsin (bR) in two different processes of retinal reconstitutions were investigated by observing the (13)C and (15)N solid-state NMR spectra of [1-(13)C]Val- and [(15)N]Pro-labeled bR. We found that NMR signals of the BC loop were sensitive to changes in protein structure and dynamics, from wild-type (WT) bR to bacterio-opsin (bO), regenerated bR and E1001 bR. Regenerated bR was prepared following the addition of retinal into bO obtained from photobleached WT-bR. E1001 bR was cultured from a retinal-deficient strain termed E1001 following the addition of retinal to growing cells. (15)N NMR signal at Pro70 in the BC loop in WT-bR was observed at 122.4 p.p.m., whereas signals were not apparent or partly suppressed in bO and regenerated bR, respectively. Similarly, the (13)C NMR signal at Val69 in the BC loop at 172.0 p.p.m. that was observed in WT-bR was significantly decreased in both regenerated bR and bO. These results suggest that the dynamic structure of the BC loop in bO was substantially altered following the removal of retinal. As a consequence, the correct protein structure failed to be recovered via the regenerating process of retinal to bO. On the other hand, (13)C and (15)N NMR signals at the BC loop in E1001 bR appeared at positions identical to those of WT-bR. The results of the current study indicate that the BC loop may not always fold correctly in the regenerated bR, which leads to different properties in the regenerated bR compared to that of WT-bR.

Self-potential variations were measured to estimate the magnitude of electrokinetic and hydrological parameters (zeta potential and permeability) of the Nojima Fault zone in Awaji, Japan. The study observed self-potential variations that seemed to be associated with water flow from the injection well to the fracture zone, which were induced by turning the injection on and off. Amplitudes of the variations were a few to 0.03 V across 320–450 m dipoles. These variations can be explained well with an electrokinetic model. The quantity k/ζ (permeability/zeta potential) is in the range 1.6 × 10−13− 5.4 × 10−13 m2/V. Permeability of the Nojima fault zone can be estimated as approximately 10−16–10−15 m2 on the assumption that the zeta potential is in the range –0.01 to –0.001 V.

Pharaonis phoborhodopsin (ppR or sensory rhodopsin II) is a negative phototaxis receptor of Natronomonas pharaonis, and forms a complex, which transmits the photosignal into cytoplasm, with its cognate transducer (pHtrII). We examined a possible local dynamics change of ppR and its D75N mutant complexed with pHtrII, using solid-state (13)C NMR of [3-(13)C]Ala- and [1-(13)C]Val-labeled preparations. We distinguished Ala C(beta) (13)C signals of relatively static stem (Ala221) in the C-terminus of the receptors from those of flexible tip (Ala228, 234, 236 and 238), utilizing a mutant with truncated C-terminus. The local fluctuation frequency at the C-terminal tip was appreciably decreased when ppR was bound to pHtrII, while it was increased when D75N, that mimics the signaling state because of disrupted salt bridge between C and G helices prerequisite for the signal transfer, was bound to pHtrII. This signal change may be considered with the larger dissociation constant of the complex between pHtrII and M-state of ppR. At the same time, it turned out that fluctuation frequency of cytoplasmic portion of pHtrII is lowered when ppR is replaced by D75N in the complex with pHtrII. This means that the C-terminal tip partly participates in binding with the linker region of pHtrII in the dark, but this portion might be released at the signaling state leading to mutual association of the two transducers in the cytoplasmic regions within the ppR/pHtrII complex.

We have so far demonstrated that well-resolved and site-specifically assigned (13)C peaks as recorded by site-directed NMR study on (13)C-labeled membrane proteins can serve as a convenient probe to reveal their local conformation and dynamics. We attempted here to clarify the extent to which (13)C NMR spectra of (13)C-labeled fully hydrated bacteriorhodopsin (bR) as a typical membrane protein are visible or well resolved in the presence of inherent fluctuation motions with frequency of 10(2)-10(8) Hz, especially at the membrane surfaces. Accordingly, we estimated the relative proportion of (13)C NMR signals from the surface areas with and without peak suppression by the accelerated transverse relaxation effect by surface-bound Mn(2+) ions, which could be effective for residues within 8.7 angstroms of the membrane surface. It turned out that the experimental findings are consistent with the predicted amount of amino acid residues under consideration located within 8.7 angstroms of the surface for [1-(13)C]Val- and Ile-labeled bR and also [3-(13)C]Ala-bR. In contrast, (13)C NMR peaks from such surfaces area are almost completely or partially suppressed for [1-(13)C]Gly-, Ala-, Leu-, Phe- and Trp-labeled bR, as a result of plausible interference of the fluctuation frequency with frequency of magic angle spinning (10(4) Hz). We further assigned several (13)C NMR signals of [1-(13)C] Val-, Trp- and Ile-labeled bR on the basis of a variety of site-directed mutants with reference to those of the wild type. Further, we recorded the (13)C NMR of bR in lipid bilayers to search for the optimal conditions to be able to obtain signals with the highest peak intensities and spectral resolution. Backbone dynamics turn out to be essential for recording (13)C NMR spectra so as to escape from motional frequencies of the order of 10(4)-10(5) Hz, either in the direction of accelerated fluctuation or slowed motions in the direction of forming the 2D array.

We have recorded (13)C nuclear magnetic resonance (NMR) spectra of [3-(13)C]Ala, [1-(13)C]Val-labeled pharaonis phoborhodopsin (ppR or sensory rhodopsin II) incorporated into egg PC (phosphatidylcholine) bilayer, by means of site-directed high-resolution solid-state NMR techniques. Seven (13)C NMR signals from transmembrane alpha-helices were resolved for [3-(13)C]Ala-ppR at almost the same positions as those of bacteriorhodopsin (bR), except for the suppressed peaks in the loop regions in spite of the presence of at least three Ala residues. In contrast, (13)C NMR signals from the loops were visible from [1-(13)C]Val-ppR but their peak positions of the transmembrane alpha-helices are not always the same between ppR and bR. The motional frequency of the loop regions in ppR was estimated as 10(5) Hz in view of the suppressed peaks from [3-(13)C]Ala-ppR due to interference with proton decoupling frequency. We found that conformation and dynamics of ppR were appreciably altered by complex formation with a cognate truncated transducer pHtr II (1-159). In particular, the C-terminal alpha-helix protruding from the membrane surface is involved in the complex formation and subsequent fluctuation frequency is reduced by one order of magnitude.

Background: Adenine paired with 8-hydroxyguanine, a major oxidatively damaged DNA lesion, is excised by mutY homologue (MYH) base excision repair protein in human cells. Since genetic polymorphisms of DNA repair genes associated with the activities and the expression levels of their products may modulate cancer susceptibility of individuals, we investigated the effect of a single nucleotide polymorphism (SNP) in the MYH gene on the difference in the expression levels of its products. Results: An aberrant size of the β type nuclear form transcript was detected in a lung cancer cell line, VMRC-LCD, by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. The transcript contained the intron 1 sequence, and it was due to alternative splicing resulting from IVS1+5G/C SNP. The presence of the upstream open reading frame (ORF) on the 5′-side of the native ORF in the β type transcript from the IVS1+5C allele could reduce the translation efficiency of the transcript into the nuclear form protein. Thus, expression vectors bearing the 5′-untranslated region sequence of either the IVS1+5G or 5C allele were constructed. In vitro translation analysis, as well as Western blot and quantitative RT-PCR analyses of the H1299 lung cancer cell line transfected with these vectors, revealed that the translation efficiency of the IVS1+5C transcript into MYH protein was much lower (≈ 30%) than that of the IVS1+5G transcript. Conclusions: The SNP at the splice donor site of the MYH gene resulted in reduced translation efficiency of its transcripts. This is the fourth case of single nucleotide variations that cause alterations in translation initiation sites and translation efficiencies in human cells.

We have examined the (13)C-NMR spectra of [3-(13)C] Ala-labeled bacteriorhodopsin and its mutants by varying a variety of environmental or intrinsic factors such as ionic strength, temperature, pH, truncation of the C-terminal alpha helix, and site-directed mutation at cytoplasmic loops, in order to gain insight into a plausible surface structure arising from the C-terminal alpha helix and loops. It is found that the surface structure can be characterized as a complex stabilized by salt bridges or metal-mediated linkages among charged side chains. The surface complex in bacteriorhodopsin is most pronounced under the conditions of 10 mM NaCl at neutral pH but is destabilized to yield relaxed states when environmental factors are changed to high ionic strength, low pH and higher temperature. These two states were readily distinguished by associated spectral changes, including suppressed (cross polarization-magic angle spinning NMR) or displaced (upfield) (13)C signals from the C-terminal alpha helix, or modified spectral features in the loop region. It is also noteworthy that such spectral changes, when going from the complexed to relaxed states, occur either when the C-terminal alpha helix is deleted or site-directed mutations were introduced at a cytoplasmic loop. These observations clearly emphasize that organization of the cytoplasmic surface complex is important in the stabilization of the three-dimensional structure at ambient temperature, and subsequently plays an essential role in biological functions.

2-(3,4,5,6-Tetrafluoro-2-hydroxyphenyl)benzoxazole (2) emits the long wavelength fluorescence around 500 nm in nonpolar solvent via the intramolecular proton transfer process in the excited state of 2 (enol-form) and also emits the intermediate wavelength fluorescence around 440 nm in polar solvent, which is assumed to originate from the excited state of 2 (anion). The ease of formation of 2 (anion), compared to 2-(2-hydroxyphenyl)benzoxazole (1), is explained by the strongly inductive fluorine atoms. In a solvent with the intermediate polarity, 2 emits both fluorescences and their relative intensity is dependent on the concentration of 2, which is supposed to be caused by the high sensitivity of the intermediate wavelength emission to the concentration quenching.

Cs5[Ru(H2O)SiW11O39] (I) and Cs5[Ru(H2O)GeW11O39] (II) are robust catalysts for water oxidation to generate oxygen with (NH4)2[Ce(NO3)6] (III) as a one-electron oxidant in water.

Case: 62-year-old female. Drainage of abscess and hysterectomy were performed for pelvic peritonitis. But stool-like exudates was observed at postoperative 2 days. Transverse colostomy and drainage of the abscess were performed. Transvaginal closure of the vaginal stump was performed for rectovaginal fistula. After a while, intractable rectovesical fistula was newly observed. Clinical course: Because patient was contracted articular rheumatism, she wanted to avoid operation. Under sufficient informed consent, we started the treatment of injection with bFGF via the drainage tube to rectovesical fistula (50 μg/day). Narrowness of the fistula was checked by fistulography, and decrease of eduction of stool like exudates was observed. The fistula was closed after 2.5 months from beginning of the treatment. Summary: We reported a case that bFGF was available for intractable nontumorous enterogenic fistula.

The oxidation of naphthalenes and methylbenzenes to quinones with aqueous 60% hydrogen peroxide in the presence of a 0.24% Pd(II)-sulfonated polystyrene-type resin catalyst was studied. The presence of electron-donating substituents on carbon-2 of naphthalenes accelerated the oxidation. A reaction path by way of hydroxylated intermediates to the quinones is proposed.By using the MINDO/3 method, quantum chemical indices such as the superdelocalizability for electrophilic species (SEr) and for radical species (SRr), and the net charge (Qr) of naphthalenes, methylbenzenes, and naphthols were calculated. The activity in the above reaction could be explained in terms of bothe Qr and SEr. The decrease of the selectivity was correlated with side reactions by radical species attacking the sites of largest SRr.

The oxidation of 2-methylnaphthalene to 2-methyl-1, 4-naphthoquinone (vitamin K3) was carried out with aqueous (60%) H2O2 in acetic acid in the presence of Pd(II) salts at 50°C. The activities of Pd(II) salts were in the order of SO2-4>ClO-4>NO-3>AcO->PO3-4>Cl->Br->I-. The Pd(II) salts such as PdSO4, Pd(ClO4)2, Pd(AcO)2, and PdCl2 were heterogenized by ion exchange between Pd(II) ion and protons of sulfonated polystyrene-type resin (SP resin) to give Pd(II)-SP resin. The use of 0.24% Pd(II)-SP resins gave a good yield (50 to 60%) of vitamin K3 at 50°C. The catalyst could be recovered by filtration and reused conveniently.