Simultaneous isolation of two species, Brachyspira pilosicoli and Brachyspira aalborgi, from a patient with ulcerative colitis.

We succeeded in the simultaneous isolation of Brachyspira (B.) aalborgi and B. pilosicoli from a patient with ulcerative colitis. B. pilosicoli grew quickly and formed colonies within 7 days, while the growth of B. aalborgi was very slow and took over 21 days. Simultaneous isolation of B. pilosicoli and B. aalborgi from a common specimen is generally recognized to be difficult, mainly due to differences in their growth requirements and the growth rates. However, we succeeded in isolating both species from a patient with ulcerative colitis and this is first evidence. The present results suggest that ulcerative colitis may be caused by simultaneous infection with B. pilosicoli and B. aalborgi.

Symmetry Transition Preserving Chirality in QCD: A Versatile Random Matrix Model.

We consider a random matrix model which interpolates between the chiral Gaussian unitary ensemble and the Gaussian unitary ensemble while preserving chiral symmetry. This ensemble describes flavor symmetry breaking for staggered fermions in 3D QCD as well as in 4D QCD at high temperature or in 3D QCD at a finite isospin chemical potential. Our model is an Osborn-type two-matrix model which is equivalent to the elliptic ensemble but we consider the singular value statistics rather than the complex eigenvalue statistics. We report on exact results for the partition function and the microscopic level density of the Dirac operator in the ϵ regime of QCD. We compare these analytical results with Monte Carlo simulations of the matrix model.

Chemokine ligand 2/chemokine receptor 2 signaling in the trigeminal ganglia contributes to inflammatory hyperalgesia in rats.

This study investigated the functional significance of hyperalgesia in the CCL2/CCR2 signaling system in trigeminal ganglion (TG) neurons following inflammation. Inflammation was induced by injection of complete Freund's adjuvant (CFA) into the whisker pad of rats. The escape threshold from mechanical stimulation applied to the whisker pad 2days later was significantly lower in CFA-treated rats than in naïve rats. Fluorogold (FG) labeling was used to identify the TG neurons innervating the whisker pad. FG-labeled TG neurons were immunoreactive for CCL2/CCR2. The mean number of CCL2/CCR2-immunoreactive small/medium-diameter TG neurons was significantly higher in inflamed rats than in naïve rats. Using whole-cell patch-clamp experiments in small-diameter TG neurons, the threshold current of FG-labeled TG neurons in inflamed rats was significantly decreased compared to naïve rats. The number of spike discharges during current injections by FG-labeled TG neurons in inflamed rats was significantly increased compared to naïve rats. These characteristic effects were abolished by co-application of a CCL2 receptor antagonist. The present study provides evidence that CCL2 enhances the excitability of small-diameter TG neurons following facial skin inflammation via the upregulation of CCR2. These findings suggest that ganglionic CCL2/CCR2 signaling is a therapeutic target for the treatment of trigeminal inflammatory hyperalgesia.

Drug-susceptibility of isolates of Brachyspira hyodysenteriae isolated from colonic mucosal specimens of pigs collected from slaughter houses in Japan in 2009.

Twenty nine isolates identified as Brachyspira hyodysenteriae were most susceptible to carbadox and metronidazole, whereas they were resistant to macrolides. The isolates showed intermediate susceptibility to tiamulin, lincomycin, penicillin G, ampicillin, chloramphenicol, tetracycline, enrofloxacin and valnemulin, with MIC50 values ranging from 0.39 to 3.13.

Expression of transient receptor potential vanilloid 1 and anoctamin 1 in rat trigeminal ganglion neurons innervating the tongue.

Transient receptor potential vanilloid 1 (TRPV1) is a polymodal sensor that is activated by heat (>43 °C), acid, or capsaicin, the pungent ingredient of hot peppers. Reports that mice lacking TRPV1 display heat avoidance behaviors and TRPV1-negative neurons respond to heat suggest that an additional heat sensor is present. Anoctamin 1 (ANO1; also known as transmembrane protein 16A [TMEM16A]), is a component of Ca(2+)-activated chloride channels (CaCCs), and has been recently identified as a heat sensor, activated by temperatures over 44 °C. ANO1 is highly co-localized with TRPV1 in small-diameter dorsal root ganglion (DRG) neurons. The aim of the present study was to investigate co-expression of ANO1 and TRPV1 in rat trigeminal ganglion (TG) neurons innervating the tongue by using retrograde labeling and immunohistochemical techniques. Fluoro-gold (FG) retrograde labeling was used to identify the TG neurons innervating the anterior two thirds of the tongue; as expected, most labeling was detected in the mandibular division of the TGs. The FG-labeled TG neurons showed TRPV1 immunoreactivity (17.9%) and ANO1 immunoreactivity (13.7%), indicating that TRPV1- and ANO1-expressing neurons were present in the mandibular division of the TGs. Seventy-six percent of the ANO1-immunoreactive TG neurons were also immunoreactive for TRPV1; this co-expression was mainly detected in small- to medium-diameter TG neurons. The high degree of co-expression of TRPV1 and ANO1 suggests that cooperation between ANO1 and TRPV1 plays a role in the signaling pathways of nociceptive TG neurons.

Brain-derived neurotrophic factor enhances the excitability of small-diameter trigeminal ganglion neurons projecting to the trigeminal nucleus interpolaris/caudalis transition zone following masseter muscle inflammation.

BACKGROUND: The trigeminal subnuclei interpolaris/caudalis transition zones (Vi/Vc) play an important role in orofacial deep pain, however, the role of primary afferent projections to the Vi/Vc remains to be determined. This study investigated the functional significance of hyperalgesia to the brain-derived neurotrophic factor (BDNF)-tyrosine kinase B (trkB) signaling system in trigeminal ganglion (TRG) neurons projecting to the Vi/Vc transition zone following masseter muscle (MM) inflammation. RESULTS: The escape threshold from mechanical stimulation applied to skin above the inflamed MM was significantly lower than in naïve rats. Fluorogold (FG) labeling was used to identify the TRG neurons innervating the MM, while microbeads (MB) were used to label neurons projecting to the Vi/Vc region. FG/MB-labeled TRG neurons were immunoreactive (IR) for BDNF and trkB. The mean number of BDNF/trkB-IR small/medium-diameter TRG neurons was significantly higher in inflamed rats than in naïve rats. In whole-cell current-clamp experiments, the majority of dissociated small-diameter TRG neurons showed a depolarization response to BDNF that was associated with spike discharge, and the concentration of BDNF that evoked a depolarizing response was significantly lower in the inflamed rats. In addition, the relative number of BDNF-induced spikes during current injection was significantly higher in inflamed rats. The BDNF-induced changes in TRG neuron excitability was abolished by tyrosine kinase inhibitor, K252a. CONCLUSION: The present study provided evidence that BDNF enhances the excitability of the small-diameter TRG neurons projecting onto the Vi/Vc following MM inflammation. These findings suggest that ganglionic BDNF-trkB signaling is a therapeutic target for the treatment of trigeminal inflammatory hyperalgesia.

Suppression of ATP-induced excitability in rat small-diameter trigeminal ganglion neurons by activation of GABAB receptor.

The aim of the present study was to investigate whether a GABAB receptor agonist could modulate ATP-activated neuronal excitability of nociceptive TRG neurons using perforated whole-cell patch-clamp and immunohistochemical techniques. Immunohistochemical analysis revealed that 86% of P2X3 receptor-immunoreactive, small-diameter TRG neurons co-expressed GABAB receptor. Under voltage-clamp conditions (Vh=-60mV), application of ATP activated the inward current in acutely isolated rat TRG neurons in a dose-dependent manner (10-50 µM) and this current could be blocked by pyridoxal-phosphate-6-azophenyl-27,47-disulfonic acid (PPADS) (10 µM), a selective P2 purinoreceptor antagonist. The peak amplitude of ATP-activated currents was significantly inhibited after application of GABAB receptor agonist, baclofen (10-50 µM), in a concentration-dependent and reversible manner. The baclofen-induced inhibition of ATP-activated current was abolished by co-application of 3-amino-2 (4-chlorophenyl)-2hydroxypropysufonic acid) saclofen, a GABAB receptor antagonist (50 µM). Under current-clamp conditions, application of 20 µM ATP significantly depolarized the membrane potential resulting in increased mean action potential frequencies, and these ATP-induced effects were significantly inhibited by baclofen and these effects were antagonized by co-application of saclofen. Together, the results suggested that GABAB receptor activation could inhibit the ATP-induced excitability of small-diameter TRG neurons activated through the P2X3 receptor. Thus, the interaction between P2X3 and GABAB receptors of small-diameter TRG neuronal cell bodies is a potential therapeutic target for the treatment of trigeminal nociception.

Biochemical and physiological characterization of a BLUF protein-EAL protein complex involved in blue light-dependent degradation of cyclic diguanylate in the purple bacterium Rhodopseudomonas palustris.

Organisms adapt their physiologies in response to the quality and quantity of environmental light. Members of a recently identified photoreceptor protein family, BLUF domain proteins, use a flavin chromophore to sense blue light. Herein, we report that PapB, which contains a BLUF domain, controls the biofilm formation of the purple photosynthetic bacterium Rhodopseudomonas palustris. Purified PapB undergoes a typical BLUF-type photocycle, and light-excited PapB enhances the phosphodiesterase activity of the EAL domain protein, PapA, which degrades the second messenger, cyclic dimeric GMP (c-di-GMP). PapB directly interacts with PapA in vitro in a light-independent manner and induces a conformational change in the preformed PapA-PapB complex. A PapA-PapB docking simulation, as well as a site-directed mutagenesis study, identified amino acids partially responsible for the interaction between the PapA EAL domain and the two C-terminal α-helices of the PapB BLUF domain. Thus, the conformational change, which involves the C-terminal α-helices, transfers the flavin-sensed blue light signal to PapA. Deletion of papB in R. palustris enhances biofilm formation under high-intensity blue light conditions, indicating that PapB functions as a blue light sensor, which negatively regulates biofilm formation. These results demonstrate that R. palustris can control biofilm formation via a blue light-dependent modulation of its c-di-GMP level by the BLUF domain protein, PapB.

Dense QCD in a finite volume.

We study the properties of QCD at high baryon density in a finite volume where color superconductivity occurs. We derive exact sum rules for complex eigenvalues of the Dirac operator at a finite chemical potential, and show that the Dirac spectrum is directly related to the color superconducting gap Delta. Also, we find a characteristic signature of color superconductivity: an X-shaped spectrum of partition function zeros in the complex quark mass plane near the origin, reflecting the Z(2)LxZ(2)R symmetry of the diquark pairing. Our results are universal in the domain Delta(-1)<

Comparison of the hemagglutination of formalinized American channel catfish (Ictalurus punctatus) erythrocytes between formalinized and live Aeromonas hydrophila isolated from the catfish rearing in Kasumigaura lake in Japan.

Formalinized Aeromonas (A.) hydrophila agglutinated loosely with the formalinized American channel catfish erythrocytes (FACCE), while live A. hydrophila agglutinated tightly with the FACCE. There was a significant difference on the number of attaching bacterial cells to the FACCE (p<0.01) (n=40 erythrocytes) between formalinized and live A. hydrophila. The other bacteria such as Salmonella (S.) Typhimurium ST-5, Escherichia (E.) coli V-517 and Staphylococcus (S.) hyicus ATCC1249 used in this experiment did not attach the FACCE.

Immunochemical demonstration of alpha(s1)- and beta-casein in mouse mammary glands at early stages of pregnancy.

We generated monoclonal antibodies (MAbs) against mouse alpha(s1)- and beta-casein and used them to survey casein immunochemically in mammary glands of mice at peri-coitous and pregnant stages. Two MAb-producing hybridoma cells, designated MCalpha1 cell and MCbeta1 cell, were established. Each antibody, when used in Western blotting, recognized specifically mouse alpha(s1)- and beta-casein among a wide spectrum of proteins of both a lactating mammary homogenate and mouse skim milk. Immunohistochemistry revealed alpha(s1)- and beta-casein in sections of lactating mammary glands. Staining was found in substances in the lumen and cytoplasm of duct and alveolar cells, particularly in rough endoplasmic reticulum and the Golgi apparatus. Mammary glands at Days 2, 4, 6, 8, and 14 of pregnancy showed positive staining specific to both alpha(s1)- and beta-casein in the lumen and cytoplasm of duct cells, whereas the glands at estrus and Day 0 of pregnancy were positive mainly for alpha(s1)-casein. Semiquantitative Western blotting analysis of both casein components in epithelial cell fractions from glands during pregnancy confirmed that intra-epithelial alpha(s1)- and beta-casein changed during three phases, elevated from trace levels to detectable levels during initial stages of pregnancy (Days 0, 2, and 4), declined to lower levels during mid-pregnancy (Days 6 and 8), and then rose to high levels during late pregnancy (Day 14).