Screening of ATP hydrolysis by Zr(IV) and Eu(III) complexes

The dephosphorylation of adenosine triphosphate (ATP) by various zirconium (IV) and europium (III) complexes is described. Release of phosphate was followed by an automated, robotics-based colorimetric assay which is compatible with the presence of these high-valent metal ions.     

Biomass Recovery Method for Adenosine Triphosphate (ATP) Quantification Following UV Disinfection

A biomass recovery method was developed to monitor UV disinfection efficacy using adenosine triphosphate (ATP). Typically, disinfection monitoring at wastewater treatment facilities (WWTFs) involves quantifying fecal and total coliforms or colony forming units, the results of which take a minimum of 24 h to produce. ATP quantification immediately before and after UV treatment, which takes only minutes, shows little reduction and often an increase in the microbial population since UV irradiation results in cells that are viable (i.e., still producing ATP) but not culturable. To overcome this, our biomass recovery method incorporates an incubation step to encourage life cycling of microbes. Average log reductions in cellular ATP (cATP) were found to be ?0.28 ± 0.19, ?0.011 ± 0.153, ?0.17 ± 0.32, and 0.065 ± 0.074 using direct ATP measurements on UV-treated samples from WWTFs A, B, C, and D, respectively, while those using the recovery method were correspondingly 0.17 ± 0.34, 1.8 ± 0.8, 0.20 ± 0.35, and 0.72 ± 0.26. The response of the biomass recovery-ATP method indicated a significant direct correlation to the microbial population reduction observed in heterotrophic plate count (HPC) and Colilert® methods using both pure Escherichia coli culture and secondary municipal wastewater effluent.     

荧光光度法研究Eu3+-四环素络合物与三磷酸腺苷二钠(ATP)的相互作用及其分析应用

本文建立了一种荧光光度法定量测定三磷酸腺苷二钠(ATP)的新方法。利用荧光光谱和吸收光谱研究了三磷酸腺苷二钠(ATP)与四环素的相互作用,四环素(TC)与Eu3 形成二元络合物发射出铕(Ⅲ)位于612nm处的特征荧光,加入三磷酸腺苷二钠(ATP)后,体系的荧光强度进一步显著增强,且增强的荧光强度与ATP的加入量成正比,据此建立了荧光分光光度法测量ATP的方法。实验测得,ATP的线性范围是3.00×10-7~4.50×10-6mol.L-1,检出限为4.68×10-8mol.L-1,研究了共存物质的影响,该方法成功的用于样品中ATP的测定。本文还讨论了Eu3 -四环素与三磷酸腺苷二钠(ATP)的相互作用机理。     

Ursolic acid and other pentacyclic triterpenoids stimulate intestinal alkaline sphingomyelinase in vitro

Purpose: Alkaline sphingomyelinase (alk‐SMase) is an enzyme that hydrolyses sphingomyelin in a bile salt‐dependent manner in the gastrointestinal tract, and has been proposed as an inhibitor of colon carcinogenesis. Ursolic acid (UA) is a plant‐derived pentacyclic triterpenoid that has been shown to have anti‐proliferative and apoptotic effects on HT29 human colon adenocarcinoma cells, with activation of alk‐SMase as an early event. The aim of this study was to study the in vitro effects of UA and its analogues on the activity of purified rat intestinal alk‐SMase. Methods: Rat intestinal alk‐SMase activity was determined after incubation with UA in the presence and absence of taurocholate (TC). The effect was compared with boswellic acids, another group of pentacyclic triterpenoids. Results: UA enhanced the activity of rat intestinal alk‐SMase in a dose‐dependent manner, without a similar effect on bacterial neutral SMase. Four types of boswellic acid also increased the enzyme activity, with the effect of acetyl‐keto‐β‐boswellic acid being most potent. Activation of alk‐SMase by TC at a low concentration (0.4 mM), but not at a high concentration, was enhanced by UA. Conclusions: Ursolic acid and four types of boswellic acid, all pentacyclic triterpenoids, have a stimulatory effect on the activity of intestinal alk‐SMase.     

Delignification of intact biomass and cellulosic coproduct of acid‐catalyzed hydrolysis

The kinetics of acid‐catalyzed hemicellulose removal and also alkaline delignification of oat hull biomass were investigated. All three operational parameters namely, catalyst concentration (0.10–0.55 N H₂SO₄), temperature (110–130°C), and residence time (up to 150 min) affected the efficiency of hemicellulose removal, with 100% of hemicellulose removed by appropriate selection of process parameters. Analysis of delignification kinetics (in the temperature range of 30–100°C) indicated that it can be expressed very well by a two‐phase model for the crude biomass and also for the hemicellulose‐prehydrolyzed material. The application of acid‐catalyzed prehydrolysis improved the capacity of lignin dissolution especially at lower temperatures (30 and 65°C) and accelerated the dissolution of lignin. This acceleration of delignification by prehydrolysis was possible at all levels of temperature in the bulk phase; however, results were more significant at the lower temperatures in the terminal phase. © 2015 American Institute of Chemical Engineers AIChE J, 61: 1783–1791, 2015     

Steric effects in the basic hydrolysis of poly(ethylene terephthalate)

A comparative study of the reactivity of hydroxide and various alkoxide anions was made by determining percentage weight loss of poly(ethylene terephthalate) fiber. Regardless of the basicity of the anions, the order of reactivity was found to be hydroxide < tert-butoxide < sec-propoxide < methoxide < ethoxide. This observed order follows the nucleophilicity of the bases, and the relatively lower reactivity of sec-propoxide and tert-butoxide is assumed to be due to steric retardation during the equilibrium reactions.     

The different roles of metal ions and water molecule in the hydrolysis of ATP catalyzed by phenanthroline-based polyamines

The phenanthroline (Phen)-based polyaza ligands can efficiently recognize nucleotides and significantly enhance the rate of ATP hydrolysis under the regulation of metal ions, which act as “messenger” between the substrates and the receptors. The water molecule (or –OH) participates in the catalytic hydrolysis of ATP in different steps with different functions.     

Review of the hydrolysis of iron(III) and the crystallization of amorphous iron(III) hydroxide hydrate

Hydrolysis of ferric solutions leads initially to mono- and dinuclear species which interact to produce further species of higher nuclearity. These polynuclear species age eventually to either crystalline compounds or to an amorphous precipitate (amorphous iron(III) hydroxide hydrate). Amorphous iron(III) hydroxide hydrate is thermodynamically unstable and gradually transforms to α-FeO(OH) and α-Fe₂O₃. These crystalline products form by competing mechanisms and the proportion of each in the final product depends on the relative rates of formation. The master variable governing the rates at which these compounds form is pH. Other important factors are temperature and the presence of additives. Most additives retard the transformation and by suppressing formation of α-FeO(OH) lead to an increase in the amount of α-Fe₂O₃ in the product; some additives also directly promote formation of the latter compound. Metal ions can oftxen replace a proportion of Fe in the α-FeO(OH) and α-Fe₂O₃ lattices. At high enough concentrations they can induce formation of additional phases. Additives may also modify the morphology of the crystalline products.     

Thermodynamic studies on the formation of cobalt(II)-adenine nucleotides complexes

The formation of complexes of Co(II) with adenosine monophosphoric (AMP) and adenosine diphosphoric (ADP) anions have been studied by pH-stat and pH variable potentiometry and also by calorimetryThe existence of complexes of type ML (CoAMP and CoADP^?, MHL (CoHADP) and ML₂ [Co(ADP)^4-₂] is in good agreement with the experimental results. The stability constants of formation of these complexes as well as their corresponding thermodynamic values, obtained in an 0.2 M solution of tetramethylammonium bromide, are presented in the Tables 1, 2 and 3.     

水解(酸化)反应器在工程应用中的研究与展望

在工业废水处理中,作为预处理单元的水解(酸化)反应器,不但降低了废水的CODCr,而且提高了废水的可生化性。作者论述了水解(酸化)反应器的特点及原理;介绍了水解(酸化)反应器的类型及其在工程应用中的效果;讨论了影响水解(酸化)反应器运行的主要因素及其设计要点;展望了水解(酸化)反应器的应用前景及研究领域。     

Expression Pattern of Purinergic Signaling Components in Colorectal Cancer Cells and Differential Cellular Outcomes Induced by Extracellular ATP and Adenosine

The purine nucleotide adenosine triphosphate (ATP) is known for its fundamental role in cellular bioenergetics. However, in the last decades, different works have described emerging functions for ATP, such as that of a danger signaling molecule acting in the extracellular space on both tumor and stromal compartments. Beside its role in immune cell signaling, several studies have shown that high concentrations of extracellular ATP can directly or indirectly act on cancer cells. Accordingly, it has been reported that purinergic receptors are widely expressed in tumor cells. However, their expression pattern is often associated with contradictory cellular outcomes. In this work, we first investigated gene expression profiles through “RNA-Sequencing” (RNA Seq) technology in four colorectal cancer (CRC) cell lines (HT29, LS513, LS174T, HCT116). Our results demonstrate that CRC cells mostly express the A2B, P2X4, P2Y1, P2Y2 and P2Y11 purinergic receptors. Among these, the P2Y1 and P2Y2 coding genes are markedly overexpressed in all CRC cells compared to the HCEC-1CT normal-like colonic cells. We then explored the cellular outcomes induced by extracellular ATP and adenosine. Our results show that in terms of cell death induction extracellular ATP is consistently more active than adenosine against CRC, while neither compound affected normal-like colonic cell survival. Intriguingly, while for the P2Y2 receptor pharmacological inhibition completely abolished the rise in cytoplasmic Ca²⁺ observed after ATP exposure in all CRC cell lines, Ca²⁺ mobilization only impacted the cellular outcome for HT29. In contrast, non-selective phosphodiesterase inhibition completely abolished the effects of extracellular ATP on CRC cells, suggesting that cAMP and/or cGMP levels might determine cellular outcome. Altogether, our study provides novel insights into the characterization of purinergic signaling in CRC.     

Thermodynamic studies on formation of Europium(III)-adenine nucleotides complexes

The following equilibrium constants have been determined by constant pH potentiometric measurements in water tetramethylammonium bromide (0.2 M) medium, for the temperature range between 5°C to 35°C:

(A represents the nucleotides which can be AMP, ADP or ATP). From the effects of temperature on these equilibria we calculate their associated thermodynamic quantities. The solubility of the Europium(III) hydroxyde has also been determined under the same experimental conditions.     

Development of Potent Adenosine Monophosphate Activated Protein Kinase (AMPK) Activators

Previously, we reported the identification of a thiazolidinedione‐based adenosine monophosphate activated protein kinase (AMPK) activator, compound 1 (N‐[4‐({3‐[(1‐methylcyclohexyl)methyl]‐2,4‐dioxothiazolidin‐5‐ylidene}methyl)phenyl]‐4‐nitro‐3‐(trifluoromethyl)benzenesulfonamide), which provided a proof of concept to delineate the intricate role of AMPK in regulating oncogenic signaling pathways associated with cell proliferation and epithelial–mesenchymal transition (EMT) in cancer cells. In this study, we used 1 as a scaffold to conduct lead optimization, which generated a series of derivatives. Analysis of the antiproliferative and AMPK‐activating activities of individual derivatives revealed a distinct structure–activity relationship and identified 59 (N‐(3‐nitrophenyl)‐N′‐{4‐[(3‐{[3,5‐bis(trifluoromethyl)phenyl]methyl}‐2,4‐dioxothiazolidin‐5‐ylidene)methyl]phenyl}urea) as the optimal agent. Relative to 1 , compound 59 exhibits multifold higher potency in upregulating AMPK phosphorylation in various cell lines irrespective of their liver kinase B1 (LKB1) functional status, accompanied by parallel changes in the phosphorylation/expression levels of p70S6K, Akt, Foxo3a, and EMT‐associated markers. Consistent with its predicted activity against tumors with activated Akt status, orally administered 59 was efficacious in suppressing the growth of phosphatase and tensin homologue (PTEN)‐null PC‐3 xenograft tumors in nude mice. Together, these findings suggest that 59 has clinical value in therapeutic strategies for PTEN‐negative cancer and warrants continued investigation in this regard.     

Acidic and basic hydrolysis of poly(N‐vinylformamide)

Poly(N‐vinylformamide) (PNVF) was synthesized and hydrolyzed to poly(vinylamine) (PVAm) in both HCl and NaOH solutions. The hydrolysis kinetics and the equilibrium hydrolysis were examined experimentally at different temperatures, polymer concentrations, and acid‐ or base‐to‐amide molar ratios. The hydrolysis kinetics strongly depended on temperature, polymer, and HCl or NaOH concentrations, but showed little dependence on PNVF molecular weight. The acid hydrolysis of PNVF exhibited limited conversions because of the electrostatic repulsion among the cationic amine groups generated during hydrolysis and proton hydrates. In the basic hydrolysis, complete amide conversions were observed when the NaOH/amide molar ratios were greater than unity. The effects of temperature and PNVF concentration on the equilibrium amide conversion appeared to be negligible in both acidic and basic hydrolysis. The equilibrium conversions of base hydrolysis were higher than those of acidic hydrolysis under the same reaction conditions. At NaOH/amide ratios of less than unity, the equilibrium hydrolysis experiments revealed that one base molecule could induce the hydrolysis of more than one amide group. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 86: 3412–3419, 2002     

The discovery of compounds that stimulate the activity of kallikrein-related peptidase 3 (KLK3)

Kallikrein-related peptidase 3 (KLK3), also known as prostate-specific antigen (PSA), is the most useful biomarker for prostate cancer (PCa). KLK3 is suggested to play a role in regulating cancer growth through anti-angiogenic activity in vivo and in vitro. This feature, together with its specificity for prostate tissue, makes KLK3 an intriguing target for the design of new therapies for PCa. 3D pharmacophores for KLK3-stimulating compounds were generated based on peptides that bind specifically to KLK3 and increase its enzymatic activity. As a result of pharmacophore-based virtual screening, four small, drug-like compounds with affinity for KLK3 were discovered and validated by capillary differential scanning calorimetry. One of the compounds also stimulated the activity of KLK3, and is therefore the first published small molecule with such an activity.     

Concentration effects in the base-catalyzed hydrolysis of oligo(ethylene glycol)- and amine-containing methacrylic monomers

Concentration effects in the base-catalyzed hydrolysis of water-soluble methacrylates (3-(N,N-dimethylaminoethyl) methacrylate (DMAEMA), 2-hydroxyethyl methacrylate (HEMA) and oligo(ethylene glycol) methacrylates (OEGMAs)) have been studied. These monomers are rapidly hydrolyzed in the presence of bases at the room temperature in dilute aqueous solutions, but the reaction rate decreases sharply in highly concentrated solutions. A clear correlation was found between a form of the viscosity isotherm for DMAEMA solutions and the concentration dependence of the autocatalytic hydrolysis rate which indicates the connection of process kinetics with the structure of solutions. These data should be considered when carrying out homo- and copolymerization of the previously mentioned monomers in aqueous solutions.     

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.