Supramolecular Zn(II)-Dipicolylamine-Azobenzene-Aminocyclodextrin-ATP Complex: Design and ATP Recognition in Water

Cyclodextrins (CyDs) are water-soluble host molecules possessing a nanosized hydrophobic cavity. In the realm of molecular recognition, this cavity is used not only as a recognition site but also as a reaction medium, where a hydrophobic sensor recognizes a guest molecule. Based on the latter concept, we have designed a novel supramolecular sensing system composed of Zn(II)-dipicolylamine metal complex-based azobenzene (1-Zn) and 3A-amino-3A-deoxy-(2AS,3AS)-γ-cyclodextrin (3-NH₂-γ-CyD) for sensing adenosine-5′-triphosphate (ATP). 1-Zn showed redshifts in the UV-Vis spectra and induced circular dichroism (ICD) only when both ATP and 3-NH₂-γ-CyD were present. Calculations of equilibrium constants indicated that the amino group of 3-NH₂-γ-CyD was involved in the formation of supramolecular 1-Zn/3-NH₂-γ-CyD/ATP. The Job plot of the ICD spectral response revealed that the stoichiometry of 1-Zn/3-NH₂-γ-CyD/ATP was 2:1:1. The pH effect was examined and 1-Zn/3-NH₂-γ-CyD/ATP was most stable in the neutral condition. The NOESY spectrum suggested the localization of 1-Zn in the 3-NH₂-γ-CyD cavity. Based on the obtained results, the metal coordination interaction of 1-Zn and the electrostatic interaction of 3-NH₂-γ-CyD were found to take place for ATP recognition. The “reaction medium approach” enabled us to develop a supramolecular sensing system that undergoes multi-point interactions in water. This study is the first step in the design of a selective sensing system based on a good understanding of supramolecular structures.     

Optical Control of Cellular ATP Levels with a Photocaged Adenylate Kinase

We have developed a new tool for the optical control of cellular ATP concentrations with a photocaged adenylate kinase (Adk). The photocaged Adk is generated by substituting a catalytically essential lysine with a hydroxycoumarin-protected lysine through site-specific unnatural amino acid mutagenesis in both E. coli and mammalian cells. Caging of the critical lysine residue offers complete suppression of Adk's phosphotransferase activity and rapid restoration of its function both in vitro and in vivo upon optical stimulation. Light-activated Adk renders faster rescue of cell growth than chemically inducible expression of wild-type Adk in E. coli as well as rapid ATP depletion in mammalian cells. Thus, caging Adk provides a new tool for direct conditional perturbation of cellular ATP concentrations thereby enabling the investigation of ATP-coupled physiological events in temporally dynamic contexts.     

基于近似测地线的分层次自动铺带轨迹规划方法

自动铺带技术是一种增量制造技术，适用于制造翼面、壁板等大尺寸、小曲率复合材料构件，具有加工成本低、效率高等特点。轨迹规划是自动铺带技术的关键环节，它直接关系到复合材料构件的成型精度与质量。现有的自动铺带轨迹规划方法难以便捷兼顾调控铺放间隙与防止铺放褶皱，保证铺带产品的最终质量。有鉴于此，提出了一种基于近似测地线的分层次铺放轨迹规划方法。主要目标是合理利用预浸带允许变形能力，在防止铺放褶皱产生的前提下，保证相邻带料不产生覆盖，且间隙不超过2.5 mm。首先，研究了铺放间隙的产生与演化规律，给出了几种典型曲面上初始铺放间隙的优化方法。接下来，提出了近似测地线的数学模型与求解格式。然后，基于铺放褶皱的产生机制与铺放间隙的演化规律，确定了近似测地线的测地曲率大小与符号。最后，在双曲面模具以及自由曲面模具上验证了所提出方法的有效性。     

Visual Detection of Adenosine Triphosphate by Taylor Rising: A Simple Point-of-Care Testing Method Based on Rolling Circle Amplification

Rapid and sensitive point-of-care testing (POCT) is an extremely critical mission in practical applications, especially for rigorous military medicine, home health care, and in the third world. Here, we report a visual POCT method for adenosine triphosphate (ATP) detection based on Taylor rising in the corner of quadratic geometries between two rod surfaces. We discuss the principle of Taylor rising, demonstrating that it is significantly influenced by contact angle, surface tension, and density of the sample, which are controlled by ATP-dependent rolling circle amplification (RCA). In the presence of ATP, RCA reaction effectively suppresses Taylor-rising behavior, due to the increased contact angle, density, and decreased surface tension. Without addition of ATP, untriggered RCA reaction is favorable for Taylor rising, resulting in a significant height. With this proposed method, visual sensitive detection of ATP without the aid of other instruments is realized with only a 5 μL droplet, which has good selectivity and a low detection limit (17 nM). Importantly, this visual method provides a promising POCT tool for user-friendly molecular diagnostics.     

Comparison between lipid and protein oxidation progress in the tail and claw muscles of signal crayfish (Pacifastacus leniusculus)

Microbial activity and then oxidation progress are the most important freshness indicators during post-mortem. In this study, we monitored proteomic and microbial changes, as well as biochemical degradation, in the tail and claw muscles of crayfish stored for 12 days at +4 °C. One specified protein band at 107 kDa in the claw muscle and two specified protein bands in the tail muscle at 140 and 36–40 kDa were identified. Western blotting indicated a higher amount of oxidised proteins in the tail compared to the claw muscle. Tail muscle showed higher oxidation progress and calpain activity than claw one. Both muscles were spoiled after 12 days with respect to total viable counts. In the first days, calpain activity is the main reason for protein degradation, while protein oxidation dominates for the rest of the time. Lipid–protein oxidation progress showed probably, protein oxidation started earlier than lipid oxidation in both muscles.     

Urothelium-Specific Deletion of Connexin43 in the Mouse Urinary Bladder Alters Distension-Induced ATP Release and Voiding Behavior

Connexin43 (Cx43), the main gap junction and hemichannel forming protein in the urinary bladder, participates in the regulation of bladder motor and sensory functions and has been reported as an important modulator of day–night variations in functional bladder capacity. However, because Cx43 is expressed throughout the bladder, the actual role played by the detrusor and the urothelial Cx43 is still unknown. For this purpose, we generated urothelium-specific Cx43 knockout (uCx43KO) mice using Cre-LoxP system. We evaluated the day–night micturition pattern and the urothelial Cx43 hemichannel function of the uCx43KO mice by measuring luminal ATP release after bladder distention. In wild-type (WT) mice, distention-induced ATP release was elevated, and functional bladder capacity was decreased in the animals’ active phase (nighttime) when Cx43 expression was also high compared to levels measured in the sleep phase (daytime). These day–night differences in urothelial ATP release and functional bladder capacity were attenuated in uCx43KO mice that, in the active phase, displayed lower ATP release and higher functional bladder capacity than WT mice. These findings indicate that urothelial Cx43 mediated ATP signaling and coordination of urothelial activity are essential for proper perception and regulation of responses to bladder distension in the animals’ awake, active phase.     

一种新的三维地学空间构模方法——类三棱柱法

综合评述了目前3DGIS和3DGM领域的三维空间构建及三维拓扑关系的研究现状：进而提出了一种新的三维地学空间构模方法－－类三菱柱（ATP）法。该法采用6类基本元素和8组拓扑关系来描述和表达三维地质现象。讨论了如何利用这8组拓扑关系进行地质体查询与分析的基本思路；给出了程序实现后的3DGM实例及其任意剖面图。     

Rapid and direct estimation of active biomass on granular activated carbon through adenosine tri-phosphate (ATP) determination

Granular activated carbon (GAC) filtration is used during drinking water treatment for the removal of micropollutants such as taste and odour compounds, halogenated hydrocarbons, pesticides and pharmaceuticals. In addition, the active microbial biomass established on GAC is responsible for the removal of biodegradable dissolved organic carbon compounds present in water or formed during oxidation (e.g., ozonation and chlorination) processes. In order to conduct correct kinetic evaluations of DOC removal during drinking water treatment, and to assess the state and performance of full-scale GAC filter installations, an accurate and sensitive method for active biomass determination on GAC is required. We have developed a straight-forward method based on direct measurement of the total adenosine tri-phosphate (ATP) content of a GAC sample and other support media. In this method, we have combined flow-cytometric absolute cell counting and ATP analysis to derive case-specific ATP/cell conversion values. In this study, we present the detailed standardisation of the ATP method. An uncertainty assessment has shown that heterogeneous colonisation of the GAC particles makes the largest contribution to the combined standard uncertainty of the method. The method was applied for the investigation of biofilm formation during the start-up period of a GAC pilot-scale plant treating Lake Zurich water. A rapid increase in the biomass of up to 1.1 x 10(10)cells/g GAC dry weight (DW) within the first 33 days was observed, followed by a slight decrease to an average steady-state concentration of 7.9 x 10(9)cells/g GAC DW. It was shown that the method can be used to determine the biomass attached to the GAC for both stable and developing biofilms.     

An Estimation Method to Determine Equivalent Circuit Parameters for Harmonic Analysis

This paper proposes a new estimation method to determine arbitrary circuit parameters for the harmonic analysis of a distribution network. Harmonic analysis is important in maintaining electric power quality. An interesting phenomenon, a temporary cut‐down of the harmonic voltage level at dawn, even if demand is slightly increasing at this time, is often observed in actual power systems. This paper proposes to utilize observations of this phenomenon to estimate circuit parameters. In addition, it is claimed that a detailed equivalent circuit is preferable to a conventional simplified equivalent circuit for explaining the phenomenon. This paper also discusses the usefulness of the Harmonic Characteristics Curve, which is proposed in this paper as a new concept, at the secondary winding of the distribution transformer, and the correlation of PQ, the monitored values of the active and reactive power at the secondary winding of distribution transformer. This correlation should determine the equivalent fundamental frequency admittance of the shunt capacitors installed for distribution lines. © 2014 Wiley Periodicals, Inc. Electr Eng Jpn, 187(1): 16–23, 2014; Published online in Wiley Online Library ( wileyonlinelibrary.com ). DOI 10.1002/eej.22451