The dopamine D1 receptor is expressed and induces CREB phosphorylation and MUC5AC expression in human airway epithelium

Background Dopamine receptors comprise two subgroups, Gs protein-coupled “D1-like” receptors (D1, D5) and Gi-coupled “D2-like” receptors (D2, D3, D4). In airways, both dopamine D1 and D2 receptors are expressed on airway smooth muscle and regulate airway smooth muscle force. However, functional expression of the dopamine D1 receptor has never been identified on airway epithelium. Activation of Gs-coupled receptors stimulate adenylyl cyclase leading to cyclic AMP (cAMP) production, which is known to induce mucus overproduction through the cAMP response element binding protein (CREB) in airway epithelial cells. We questioned whether the dopamine D1 receptor is expressed on airway epithelium, and whether it promotes CREB phosphorylation and MUC5AC expression. Methods We evaluated the protein expression of the dopamine D1 receptor on native human airway epithelium and three sources of cultured human airway epithelial cells including primary cultured airway epithelial cells, the bronchial epithelial cell line (16HBE14o-), and the pulmonary mucoepidermoid carcinoma cell line (NCI-H292) using immunohistochemistry and immunoblotting. To characterize the stimulation of cAMP through the dopamine D1 receptor, 16HBE14o- cells and NCI-H292 cells were treated with dopamine or the dopamine D1 receptor agonists (SKF38393 or A68930) before cAMP measurements. The phosphorylation of CREB by A68930 in both 16HBE14o- and NCI-H292 cells was measured by immunoblot. The effect of dopamine or A68930 on the expression of MUC5AC mRNA and protein in NCI-H292 cells was evaluated by real-time PCR and immunofluorescence staining, respectively. Results The dopamine D1 receptor protein was detected in native human airway epithelium and three sources of cultured human airway epithelial cells. Dopamine or the dopamine D1-like receptor agonists stimulated cAMP production in 16HBE14o- cells and NCI-H292 cells, which was reversed by the selective dopamine D1-like receptor antagonists (SCH23390 or SCH39166). A68930 significantly increased phosphorylation of CREB in both 16HBE14o- and NCI-H292 cells, which was attenuated by the inhibitors of PKA (H89) and MEK (U0126). Expression of MUC5AC mRNA and protein were also increased by either dopamine or A68930 in NCI-H292 cells. Conclusions These results suggest that the activation of the dopamine D1 receptor on human airway epithelium could induce mucus overproduction, which could worsen airway obstructive symptoms

Pillar[6]arene acts as a biosensor for quantitative detection of a vitamin metabolite in crude biological samples

ビタミン代謝物を迅速定量できる超分子バイオセンサーを開発. 京都大学プレスリリース. 2020-12-09.Metabolic syndrome is associated with obesity, hypertension, and dyslipidemia, and increased cardiovascular risk. Therefore, quick and accurate measurements of specific metabolites are critical for diagnosis; however, detection methods are limited. Here we describe the synthesis of pillar[n]arenes to target 1-methylnicotinamide (1-MNA), which is one metabolite of vitamin B3 (nicotinamide) produced by the cancer-associated nicotinamide N-methyltransferase (NNMT). We found that water-soluble pillar[5]arene (P5A) forms host–guest complexes with both 1-MNA and nicotinamide, and water-soluble pillar[6]arene (P6A) selectively binds to 1-MNA at the micromolar level. P6A can be used as a “turn-off sensor” by photoinduced electron transfer (detection limit is 4.38 × 10−6 M). In our cell-free reaction, P6A is used to quantitatively monitor the activity of NNMT. Moreover, studies using NNMT-deficient mice reveal that P6A exclusively binds to 1-MNA in crude urinary samples. Our findings demonstrate that P6A can be used as a biosensor to quantify 1-MNA in crude biological samples

Efficient "on the fly" signature schemes based on integer factoring

In 1999, Poupard and Stern proposed on the fly signature scheme (PS-scheme), which aims at minimizing the on-line computational work for a signer. In this paper, we propose more efficient on the fly signature schemes by improving the PS-scheme. In PS-scheme, the size of secret-key is fixed by modulus n, so that this feature leads to some drawbacks in terms of both the computational work and the communication load. The main idea of our schemes is to reduce the size of secret-key in PS-scheme by using a public element g which has a specific structure. Consequently, our schemes are improved with respect to the computational work (which means the computational cost for "precomputation", "(on-line) signature generation" and "verification") and the data size such as a secret-key and a signature.Progress in cryptology - INDOCRYPT 2001 : Second International Conference on Cryptology in India, Chennai, India, December 16-20, 2001 : proceedings / C. Pandu Rangan, Cunsheng Ding (eds.)

Provably Secure Multi-signature Scheme with Signers' Intentions

In this paper, we propose a multi-signature scheme, in which each signer can express her intention in the message to be signed. An intention is a piece of information which can be attached to a signature. However, no multi-signature scheme dealing with intentions without loss of efficiency has been introduced. First, we consider a multi-signature scheme realizing the concept of signers' intentions by utilizing existing schemes, and name it primitive method. After that, we introduce the proposed multi-signature scheme which is more efficient than the primitive method in view of the computational cost for verification and in view of the signature size. The proposed multi-signature scheme is shown to be secure even against adaptive chosen message insider attacks

A multi-signature scheme with signers' intentinos secure against active attacks

In this paper, we propose a multi-signature scheme, in which each signer can express her intention associating with the message to be signed. Signers' intentions mean a kind of information which can be newly attached to a signature in signers' generating it. However, we have been introduced any multi-signature scheme dealing with intentions without loss of its efficiency. First, we consider a multi-signature scheme realizing the concept of signers' intentions by utilizing existing schemes, and name it primitive method. After that, we introduce the proposed multi-signature scheme which are more efficient in view of the computational cost for verification and in view of the signature size than the primitive method. The proposed multi-signature scheme is shown to be secure even against adaptive chosen message insider attacks.Information security and cryptology : ICISC 2001 : 4th International Conference, Seoul, Korea, December 6-7, 2001 : proceedings / Kwangjo Kim (ed.)

Determination of the freshwater origin of Coastal Oyashio Water using humic-like fluorescence in dissolved organic matter

Coastal Oyashio Water (COW), defined as a water mass with a temperature lower than 2 degrees C and a salinity lower than 33.0, is distributed in the North Pacific Ocean off southeastern Hokkaido, Japan, from winter to spring. COW is rich in macronutrients and dissolved iron and is thus considered to affect the spring phytoplankton blooms in the Oyashio region. Although river water and sea-ice melt water have been considered freshwater end-members of COW, the contributions of these freshwater sources to COW have not been well described. In this study, the humic-like components in dissolved organic matter were first applied as a parameter to evaluate the freshwater end-members of COW in March 2015. Linear regressions with negative slopes were determined between the humic-like components and the salinity of COW. The intercepts of the regressions against the humic-like components were within the ranges of those observed for the local rivers of Hokkaido but were very different from those of sea ice. These findings suggest that river water contributed to the COW observed here as a freshwater end-member, although the contribution of sea-ice melt water to COW could not be evaluated. This novel approach also highlighted two different less-saline water masses in COW. The first was characterized by a lower temperature and relatively high levels of humic-like components, while the second was higher in temperature and had higher levels of humic-like components. It is suggested that these different characteristics are due to the contributions of water from different rivers and/or different effects of sea-ice melt water