Characteristics of instantaneous wind in an urban area measured by an ultra-sonic velocimetry

This paper was reviewed and accepted by the APCWE-IX Programme Committee for Presentation at the 9th Asia-Pacific Conference on Wind Engineering, University of Auckland, Auckland, New Zealand, held from 3-7 December 2017

Effects of Proton Motive Force on the Structure and Dynamics of Bovine Cytochrome <i>c</i> Oxidase in Phospholipid Vesicles

A conventional method for reconstituting cytochrome <i>c</i> oxidase (CcO) into phospholipid vesicles (COV) has been modified to permit resonance Raman (RR) analysis in the presence and absence of proton motive force (Δμ_H⁺). The COV has an average diameter of 20 nm and contains one CcO molecule within a unified orientation with Cu_A located outside the COV. The process of generation of Δμ_H⁺ across the membrane was monitored spectrophotometrically with rhodamine123 dye. The COV exhibits a respiratory control ratio (RCR) value of >30 and is tolerant to RR measurements with 10 mW laser illumination for 60 min at 441.6 nm. Structural perturbations at the heme sites caused by incorporation into vesicles were clarified by spectral comparisons between solubilized CcO and COV. Absorption spectroscopy revealed that the rate of electron transfer from cytochrome <i>c</i> to O₂ is reduced significantly more in the presence of Δμ_H⁺ than in its absence. RR spectroscopic measurements indicate that CcO in COV in the “respiratory-controlled” state adopts a mixed-valence state (heme <i>a</i>²⁺ and heme <i>a</i>₃³⁺). This study establishes a supramolecular model system for experimentally examining the energy conversion protein machinery in the presence of Δμ_H⁺

Reversible O–O Bond Scission of Peroxodiiron(III) to High-Spin Oxodiiron(IV) in Dioxygen Activation of a Diiron Center with a Bis-tpa Dinucleating Ligand as a Soluble Methane Monooxygenase Model

The conversion of peroxodiiron­(III) to high-spin <i>S</i> = 2 oxodiiron­(IV) via reversible O–O bond scission in a diiron complex with a bis-tpa dinucleating ligand, 6-hpa, has been characterized by elemental analysis; kinetic measurements for alkene epoxidation; cold-spray ionization mass spectrometry; and electronic absorption, Mössbauer, and resonance Raman spectroscopy to gain insight into the O₂ activation mechanism of soluble methane monooxygenases. This is the first synthetic example of a high-spin <i>S</i> = 2 oxodiiron­(IV) species that oxidizes alkenes to epoxides efficiently. The bistability of the peroxodiiron­(III) and high-spin <i>S</i> = 2 oxodiiron­(IV) moieties is the key feature for the reversible O–O bond scission

Wind Tunnel Experiment to Measure Impact Forces of Wind-blown Spheres Colliding at the Ground

This paper was reviewed and accepted by the APCWE-IX Programme Committee for Presentation at the 9th Asia-Pacific Conference on Wind Engineering, University of Auckland, Auckland, New Zealand, held from 3-7 December 2017

Increased the number of T cell subsets in draining lymph nodes in EP3KO mice.

<p>Draining lymph nodes were collected the mice of 5 days after the DNFB or vehicle application mice. (a) Representative flow cytometric plots. (b) The numbers of CD44^- CD62L⁺ naïve (upper panel), CD44⁺CD62L⁺ central memory (middle panel), and CD44⁺CD62L^- effector memory (lower panel) subsets of CD4⁺ T cells (left panels) and CD8⁺ T cells (right panels) of B6 and EP3KO mice were counted and indicated by white and black bars, respectively (n=6). Each data represents the mean + SD. *<i>p</i><0.05.</p

Prostaglandin E₂-EP3 Axis in Fine-Tuning Excessive Skin Inflammation by Restricting Dendritic Cell Functions

<div><p>Prostaglandin E₂ (PGE₂) is produced in the skin and is suggested to play a role in the regulation of cutaneous immune homeostasis and responses. However, the multifaceted functions of PGE₂ continue to elude our understanding, especially because of the multiplicity of PGE₂ receptors—EP1, EP2, EP3, and EP4. While cAMP-elevating EP4 is known to activate the functions of cutaneous dendritic cells (DCs), including Langerhans cells (LCs) and dermal DCs, the role of cAMP-suppressing EP3 in this process remains unknown. Here we demonstrated that an EP3 receptor selective agonist, ONO-AE-248, inhibited chemotaxis and co-stimulatory molecule expressions of DCs <i>in vitro</i>. A suboptimal dose of antigen was sufficient to induce contact hypersensitivity in EP3-deficient mice. Intriguingly, EP3 deficiency did not impair skin inflammation at all when the antigen dose was sufficiently high. EP3 limited the functions of cutaneous DCs only when the antigen dose was low. In contrast to EP4, the observed unappreciated function of EP3 may stabilize the cutaneous DCs to halt the impetuous response to a suboptimal dose of antigen. Taken together, PGE₂-EP3 signaling is essential for fine-tuning excessive skin inflammation by restricting DC functions.</p> </div

Inhibition of migration and maturation of BMDCs by EP3 agonist (AE248).

<p>(a) mRNA Expression of the four EP subtypes in BMDCs. (b) Low dose PGE₂ attenuated the migration of BMDCs. BMDCs were treated with 0, 1, 10 and 100 pM PGE₂ and applied to a transwell. 100 ng/mL CCL21 was administrated to the lower chamber. Migrated BMDCs were identified as MHC class II⁺ CD11c⁺ subset in the lower chamber. The % input was calculated as follows: (the number of BMDCs migrated into the lower chamber)/(the number of BMDCs applied into the upper chamber) *100. (c) EP3 agonist attenuated the CCL21-induced migration of BMDCs. BMDCs were treated with 10 μM AE248 for 2 days and applied to a transwell. The migration assay was carried out the same way as previously descried. (d) Intracellular cAMP concentration. BMDCs were treated with 0.5 mM IBMX for15min prior to EP3 agonist treatment (30min). (e) The number of total (CD86^high MHC class II ^{middle to high}) and mature (CD86^high MHC class II ^high) BMDCs were counted after EP3 agonist incubation for 2 days (n=6). Each data represents the mean + SD. *<i>p</i><0.05.</p

Enhancement of cutaneous immune response in EP3KO mice.

<p>(a) B6 and EP3KO mice (n=5 per group) were sensitized with 0 (un-sensitized), 0.05 or 0.5%-DNFB and the ear swelling was measured 24 hours after challenge with 0.3%-DNFB. (b) Hematoxylin-eosin staining of ears after the challenge. Scale bar indicates 200 µm. The total histology score was calculated as the sum of four elements (inflammation, neutrophils, edema and epithelial hyperplasia). (c) IFN-γ mRNA in the draining lymph nodes of the un-sensitized and 0.05%-DNFB-sensitized mice. Draining lymph nodes were collected after the measurement of ear swelling (n=5). Each data represents the mean + SD. *<i>p</i><0.05. sens, sensitization; chall, challenge.</p

Enhanced cutaneous DC migration in EP3KO mice.

<p>(a) The number of FITC⁺ MHC class II⁺ DCs in the draining lymph nodes from EP3KO and WT mice 96 hours after application of 0.5 or 2% FITC (n=4). (b) Representative flow cytometry of lymph node cells. (c) The numbers of FITC⁺ LCs (FITC⁺ Langerin⁺ cells) or dermal DCs (FITC⁺ Langerin^- cells) in the draining lymph nodes of the mice after application of 0.5% FITC. White and black columns indicate B6 and EP3KO mice, respectively (n=4). Each data represents the mean + SD. *<i>p</i><0.05.</p

Inhibition of migration and maturation of LCs by EP3 agonist (AE248).

<p>(a) Expression of the four EP subtypes in CD11c⁺ epidermal cell suspensions (Langerhans cells) (Left; mRNA, Right; protein). (b) LCs treated with EP3 agonist for 24 hours were applied to the upper chamber of a transwell. After 3 hours incubation, the numbers of MHC class II ⁺ CD11c⁺ cells in the lower chamber including 100 ng/mL CCL21 were counted. White and black bars indicates the data from B6 and EP3KO mice, respectively (n=3). (c) Expression of maturation markers (CD80, CD86 and CD54) on LCs were analyzed by FACS 24 hours after EP3 agonist treatment (n=3). *<i>p</i><0.05. MFI, mean fluorescence intensity.</p