Value Of ATP Research Articles

ATP and WTP analysis in determining retribution rate for building construction permit in Boyolali district

Building Construction Permit (BCP) is a permit granted by the regional government except for buildings with particular government functions to build new, change, expand, reduce, and maintain buildings according to applicable administrative and technical requirements. This study aims to analyze and determine the unit price building levies for building permits in Boyolali Regency. The study aim also can fulfill the concept of cost recovery and equity. The amount of retribution paid by the community is equal to the operational costs in the licensing process or services provided by the local government and adjusted to the public ability to pay and willingness to pay or benefits provided by the local government. This research is a descriptive-analytical study. The sample in this study was 100 respondents. This research’s data sources consist of primary data in interviews and questionnaires and secondary data in operational costs and targets and realization of BCP income. The results showed that the BCP levy rate set’s value was lower than the value of the Activity Operational Costs (OAC). However, the value of the OAC tariff is greater than the value of ATP and WTP. The ability to pay BCP levies is equivalent to the willingness to pay BCP levies.

Immune cell function assays in the diagnosis of infection in pediatric liver transplantation: an open-labeled, two center prospective cohort study.

BackgroundLimited studies have been performed in assessment of immune status of pediatric liver transplants (PLTs). We conducted this study to evaluate Cylex immune cell function assay in diagnosis of infection and its potential clinical application in Chinese infant PLTs.MethodsIn this prospective cohort study, 227 infant PLTs from two medical centers were enrolled, and 216 completed the study. Cylex ATP values were measured before and after liver transplantation (LT) at week 1, 2, 3, 4, 8, 12 and 24 respectively. Accordingly, patients’ clinical records, including demographic data, liver function results, tacrolimus dosages and concentrations were collected and analyzed.ResultsOne hundred and sixty of 216 PLTs (74.1%) were diagnosed infection based on the parameters including abnormal vital signs, imaging changes, and pathogens detection, while 44 (20.4%) were clinically stable and 12 (5.6%) experienced acute rejection. The median Cylex ATP value in infant PLTs post-surgery reduced significantly in infection group compared to stable group (median, 137 vs. 269 ng/mL, P<0.001). Receiver operating characteristic (ROC) curve analysis determined that the cut-off value of Cylex ATP was 152 ng/mL in diagnosis of infection [area under the curve (AUC): 0.784, 95% CI: 0.720–0.848]. Meanwhile, Cylex ATP value showed no correlation to tacrolimus dosage, blood concentration, dose-normalized concentration/dose ratio or Kaup index. However, it tended to correlate weakly with the white blood cell (WBC) number (R =0.462, P<0.0001) and lymphocyte counts (R =0.363, P<0.0001).ConclusionsIn this study, we demonstrated that low Cylex ATP represented partly over-immunosuppression and had diagnostic value in infant PLTs with infections, which might assist individualized immunosuppression in PLT patients.

Melatonin ameliorates cisplatin-induced asthenozoospermia via reducing oxidative stress

Background and objective: Cisplatin is an alkylating agent that has become a first-line therapy for some tumors. However, overproduction of reactive oxygen species (ROS) by cisplatin can cause male infertility, which can affect patients' quality of life. Melatonin, which has the ability to provide resistance against oxidation, is a potential therapy for male infertility caused by cisplatin.Material and methods: Normal human spermatozoa samples were divided into (i) control group incubated with physiological saline solution; (ii) cisplatin group incubated with cisplatin; (iii) melatonin group incubated with melatonin; and (iv) melatonin + cisplatin group incubated with melatonin and cisplatin. Spermatozoa motility was measured using a computer-aided semen analysis system. Additionally, we determined spermatozoa apoptosis through terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling. The mitochondrial membrane potential (∆Ψm) and levels of ATP, ROS, and malondialdehyde, as a metabolite of oxidative stress, were measured to study the mechanism by which melatonin can protect human spermatozoa.Results: Melatonin improved sperm motility, and decreased apoptosis and increased the ∆Ψm value and level of ATP in sperm. More importantly, the overproduction of ROS induced by cisplatin was reduced by melatonin. Furthermore, western blotting revealed that melatonin increased the expression of antioxidant pathway components nuclear factor erythroid 2-related factor (Nrf2) and heme oxygenase-1 (HO-1) that was reduced by cisplatin.Conclusion: Our studies suggested that melatonin might protect human spermatozoa against the effects of cisplatin through Nrf2/HO-1.

Actomyosin interaction at low ATP concentrations.

In vitro motility assay (IVMA) experiments were performed to analyze the movement of actin filaments sliding on a pavement of myosin molecules at different [ATP] and [ADP]. In standard experimental conditions at [ATP]=2mM, about 80% of the actin filaments move in unloaded conditions with a constant velocity. However, a fraction of at least 20% static actin filaments is always present. The accepted explanation is the occurrence of damaged "rigor"-like myosin heads that do not undergo the normal ATP-dependent cycling motion. However, in a series of IVMA experiments performed at different [ATP] we observed that the mobility of actin filaments increased with lowering [ATP]. We investigated the influence of [ATP] on the number of mobile actin filaments. IVMA experiments were performed at controlled nucleotide concentrations and the percentage of mobile filaments accurately determined by specific operator-guided software. The value of ΔG ATP involved was determined. Results showed that the number of mobile actin filaments sliding on type 2B heavy meromyosin isoform (2B HMM) increased at very low [ATP] accompanied by less negative ΔG ATP values. Similar results were obtained by increasing [ADP]. Performing experiments at the same [ATP] with different myosin types, we found a higher number of mobile actin filaments on slow type 1 HMM with respect to type 2B HMM while the highest number of mobile actin filaments was found on single-head myosin (S1 fraction). We also found that [ATP] did not influence the percentage of mobile actin filaments sliding on S1. Our results reveal novel aspects of actomyosin interaction.

Thiazolo[3,2-a]benzimidazol-3(2H)-one derivatives: Structure–activity relationships of selective nucleotide pyrophosphatase/phosphodiesterase1 (NPP1) inhibitors

Ecto-nucleotide pyrophosphatase/phosphodiesterase1 (NPP1) is the most important member of the NPP family, which consists of seven closely related proteins (NPP1–NPP7). This glycoprotein is a membrane-associated or secreted enzyme, which catalyzes the hydrolysis of a wide range of phosphodiester bonds, e.g., in nucleoside triphosphates, dinucleotides and nucleotide sugars. NPP1 plays a crucial role in various physiological functions including bone mineralization, soft-tissue calcification, and insulin receptor signaling. Recently, an upregulated expression of NPP1 has been observed in astrocytic brain cancers. Therefore, NPP1 has been proposed as a novel drug target for the treatment of glioblastoma. Despite their therapeutic potential, only few NPP1 inhibitors have been reported to date, which are in most cases non- or only moderately selective. The best investigated NPP1 inhibitors so far are nucleotide derivatives and analogs, however they are not orally bioavailable due to their high polarity. We identified thiazolo[3,2-a]benzimidazol-3(2H)-one derivatives as a new class of NPP1 inhibitors with drug-like properties. Among the 25 derivatives investigated in the present study, 2-[(5-iodo-2-furanyl)methylene]thiazolo[3,2-a]benzimidazol-3(2H)-one (17) was found to be the most potent NPP1 inhibitor with a Ki value of 467nM versus ATP as a substrate and an un-competitive mechanism of inhibition. Compound 17 did not inhibit other human ecto-nucleotidases, including NTPDase1 (CD39), NTPDases2-3, NPP2, NPP3, tissue-nonspecific alkaline phosphatase (TNAP), and ecto-5′-nucleotidase (eN, CD73), and is thus highly selective for NPP1.

Abstract A180: Discovery of furanone derivatives as novel Cdc7 inhibitors with anti-tumor activity

Abstract Introduction&amp;gt; Cell cycle kinases have been recognized as potential anticancer targets, because loss of regulated cell cycle progression would lead to uncontrolled cell growth and development of cancer. Cdc7 kinase is involved in regulation of the cell cycle and plays important roles in DNA replication. Notably, inhibition of Cdc7 in cancer causes lethal S phase or M phase progression, whereas normal cells can survive, most likely through induction of cell cycle arrest at the DNA replication checkpoints. Therefore Cdc7 kinase has emerged as a promising therapeutic target for the treatment of cancers. Here we present a novel series of furanone derivatives with potent inhibitory effects on Cdc7 kinase. &amp;lt;Methods&amp;gt; To identify hit compounds that are insensitive or resistant to high ATP concentration, a high-throughput screening (HTS) was performed in the presence of 100 microM ATP which corresponds to 36-times higher than the Km value of ATP for Cdc7 kinase activity. To characterize inhibitors, ATP dependency and pre-incubation effects were examined. To determine the efficacy of Cdc7 inhibitors, phosphorylation of MCM2 protein in cells were analyzed by western blotting. Selected compounds were subjected to a kinase selectivity panel assay. &amp;lt;Results&amp;gt; Only a single hit compound was identified from the HTS, reflecting extremely low hit rate compared to that of other kinase targets in our past HTS. Subsequent hit to lead studies led to the identification of a novel furanone compound as a potential lead compound for further optimization study. Interestingly, these furanone derivatives showed slow-binding inhibition against Cdc7, and exhibited strong inhibitory activity for Cdc7 even in the presence of 1 mM ATP that was added after the pre-incubation of the kinase and compound. Notably this pre-incubation effect was not observed with other kinases tested. Detailed results using cancer cells will also be presented in this poster. Citation Format: Takayuki Irie, Tokiko Asami, Ayako Sawa, Chika Taniyama, Yoko Funakoshi, Yuko Uno, Hisao Masai, Masaaki Sawa. Discovery of furanone derivatives as novel Cdc7 inhibitors with anti-tumor activity. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr A180.

An Improved Method for P2X7R Antagonist Screening.

ATP physiologically activates the P2X7 receptor (P2X7R), a member of the P2X ionotropic receptor family. When activated by high concentrations of ATP (i.e., at inflammation sites), this receptor is capable of forming a pore that allows molecules of up to 900 Da to pass through. This receptor is upregulated in several diseases, particularly leukemia, rheumatoid arthritis and Alzheimer's disease. A selective antagonist of this receptor could be useful in the treatment of P2X7R activation-related diseases. In the present study, we have evaluated several parameters using in vitro protocols to validate a high-throughput screening (HTS) method to identify P2X7R antagonists. We generated dose-response curves to determine the EC50 value of the known agonist ATP and the ICs50 values for the known antagonists Brilliant Blue G (BBG) and oxidized ATP (OATP). The values obtained were consistent with those found in the literature (0.7 ± 0.07 mM, 1.3-2.6 mM and 173-285 μM for ATP, BBG and OATP, respectively). The Z-factor, an important statistical tool that can be used to validate the robustness and suitability of an HTS assay, was 0.635 for PI uptake and 0.867 for LY uptake. No inter-operator variation was observed, and the results obtained using our improved method were reproducible. Our data indicate that our assay is suitable for the selective and reliable evaluation of P2X7 activity in multiwell plates using spectrophotometry-based methodology. This method might improve the high-throughput screening of conventional chemical or natural product libraries for possible candidate P2X7R antagonist or agonist

Application value of ATP based bioluminescence tumor chemosensitivity assay in the chemotherapy for hydrothorax caused by non-small cell lung cancer

Objective The aim of the study was to investigate the clinical value and application of ATP based bioluminescence tumor chemosensitivity assay (ATP-TCA) in the chemotherapy for hydrothorax caused by non-small cell lung cancer (NSCLC).

Conserved extracellular cysteines differentially regulate the potentiation produced by Zn2+ in rat P2X4 receptors

One feature of the amino acid sequence of P2X receptors identified from mammalian species, Xenopus laevis and zebrafish is the conservation of ten cysteines in the extracellular loop. Little information is available about the role of these conserved ectodomain cysteines in the function of P2X receptors. Here, we investigated the possibility that ten conserved cysteine residues in the extracellular loop of the rat P2X4 receptor may regulate zinc potentiation of the receptor using a series of individual cysteine to alanine point mutations and functional characterization of recombinant receptors expressed in Xenopus oocytes. For the C116A, C132A, C159A, C165A, C217A and C227A mutants, 10µM zinc did not significantly affect the current activated by an EC40 concentration of ATP. By contrast, 5µM zinc shifted the ATP concentration-response curve to the right in a parallel manner for both the C261A and C270A mutants and the magnitudes of those shifts were similar to that of the wildtype receptor. Interestingly, for the C126A and C149A mutants, 5µM zinc potentiated ATP-activated current, but increased the maximal response to ATP by 90% and 81% respectively, without significantly changing the EC50 value of ATP. Thus, these results suggest that cysteines and disulfide bonds between cysteines are differentially involved in the potentiation of the rat P2X4 receptor by zinc.

Effects of wild boar (Sus scrofa) grazing on soil properties in Mediterranean environment

Abstract Two sites (Asciano and Castelfalfi) were selected in Tuscany (central-west Italy) in order to study the effects of wild boar activities (rooting and trampling) at two different densities and on the organic matter quality and functionality in soil ecosystems. In each site, two study areas with different vegetation (woodland (W) and olive tree grove (O)) were identified to evaluate the influence of vegetal cover on soil properties. Physical, chemical, biological and chemico-structural (pyrolysis–gas chromatography) analysis were combined to assess the evolution of soil characteristics. In the low boar density site (Asciano) soil organic matter quality and biological activity resulted positively affected by rooting and trampling activity (higher value of total organic carbon, total nitrogen, ATP and biomass carbon in soil grazed), while the high wild boar density (Castelfalfi) was responsible for a higher soil degradation as suggested by the lower value of chemico-physical and biological parameters. The effect of wild boar activity on soil depends also on the type of vegetation, in fact, woodland vegetation, with a greater contribution in organic matter, seemed to be more efficient in the protection of soil against degradation with respect to olive cover, also in Castelfalfi site, showing an improvement in the physical and biological soil properties.

Critical Role for CD38-mediated Ca2+ Signaling in Thrombin-induced Procoagulant Activity of Mouse Platelets and Hemostasis

CD38, a multifunctional enzyme that catalyzes the synthesis of intracellular Ca(2+) messengers, cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP), is known to be expressed on platelets. However, the role of CD38 in platelets remains unclear. Our present results show that treatment of platelets with thrombin results in a rapid and sustained Ca(2+) signal, resulting from a coordinated interplay of Ca(2+)-mobilizing messengers, inositol 1,4,5-trisphosphate, cADPR, and NAADP. By dissecting the signaling pathway using various agents, we delineated that cADPR and NAADP are sequentially produced through CD38 internalization by protein kinase C via myosin heavy chain IIA following phospholipase C activation in thrombin-induced platelets. An inositol 1,4,5-trisphosphate receptor antagonist blocked the thrombin-induced formation of cADPR and NAADP as well as Ca(2+) signals. An indispensable response of platelets relying on cytosolic calcium is the surface exposure of phosphatidylserine (PS), which implicates platelet procoagulant activity. Scrutinizing this parameter reveals that CD38(+/+) platelets fully express PS on the surface when stimulated with thrombin, whereas this response was decreased on CD38(-/-) platelets. Similarly, PS exposure and Ca(2+) signals were attenuated when platelets were incubated with 8-bromo-cADPR, bafilomycin A1, and a PKC inhibitor. Furthermore, in vivo, CD38-deficient mice exhibited longer bleeding times and unstable formation of thrombus than wild type mice. These results demonstrate that CD38 plays an essential role in thrombin-induced procoagulant activity of platelets and hemostasis via Ca(2+) signaling mediated by its products, cADPR and NAADP.

Effect of ammonium, sodium, and potassium ions on rabbit muscle phosphofructokinase-1 and adenylate kinase activities

This report shows that 30 nM PFK-1 and 30 nM AK were both affected by the presence of NH4+, Na+, and K+ salts but with opposite consequences. Low concentrations of PFK-1 lose about half of its activity as a result of dilution and become susceptible to further activity losses owing to the presence of monovalent salts. On the other hand low concentrations of AK lose about 75 percent of its activity but regains activity losses owing to the presence of monovalent salts. It was determined that regain of AK activity did not appear to be a reflection of a major effect on the Km value of either AMP or ATP. Dilution to 30 nM AK resulted in no increase Km values compared to Km values at 140 nM AK. Dilution caused major decreases in the maximum velocities, Vmax, when ATP or fructose 6-phosphate was the variable substrate. It was shown in earlier reports that these same low concentrations of PFK-1 and AK were susceptible inhibitions by ascorbate. These attributes are discussed as they may relate to the role of ascorbate facilitation glycogen synthesis in resting muscle and the role that the cytoskeleton infrastructure scaffold may play is also discussed.

Conserved extracellular cysteines differentially regulate the inhibitory effect of ethanol in rat P2X 4 receptors

Relatively little information is available about the molecular mechanism of ethanol inhibition of P2X receptors. Here, we investigated the possibility that 10 conserved cysteine residues in the extracellular loop of the rat P2X 4 receptor may regulate ethanol inhibition of the receptor using a series of individual cysteine to alanine point mutations. Each of the mutated receptors generated robust inward current in response to ATP and the mutations produced less than a sixfold change in the ATP EC 50 value. For the C116A, C126A, C149A, and C165A mutants, 100 mM ethanol did not significantly affect the current activated by an EC 40 concentration of ATP. By contrast, for the C261A and C270A mutants, ethanol inhibited ATP-activated current in a competitive manner similar to that for the wild-type receptor. Interestingly, for the C132A, C159A, C217A, and C227A mutants, ethanol inhibited ATP-activated current, but decreased the maximal response to ATP by 70–75% without significantly changing the EC 50 value of ATP, thus exhibiting a noncompetitive-type inhibition. The results suggest that cysteines and disulfide bonds between cysteines are differentially involved in the inhibition of the rat P2X 4 receptor by ethanol.

Application value of ATP based bioluminescence tumor chemo-sensitivity assay in the chemotherapy for ascites caused by recurrent ovarian cancer

Objective To investigate the clinical value and application of ATP based bioluminescence tumor chemo-sensitivity assay (ATP-TCA) in the chemotherapy for ascites caused by recurrent ovarian cancer.

ATP content and biomass activity in sequential anaerobic/aerobic reactors.

Specific ATP content of volatile solids was measured to characterize the sludge activity in a sequential an-aerobic/aerobic wastewater treatment system, with an upflow anaerobic sludge blanket (UASB) reactor and a three-phase aerobic fluidized bed (AFB) reactor. The wastewater COD level was 2000-3000 mg/L in simulation of real textile waste-water. The ATP content and the specific ATP contents of volatile solids at different heights of the UASB reactor and those of the suspended and immobilized biomass in the AFB reactor were measured. In the UASB reactor, the maximum value of specific ATP (0.85 mg ATP/g VS) was obtained at a hydraulic retention time (HRT) 7.14 h in the blanket solution. In the AFB reactor, the specific ATP content of suspended biomass was higher than that of immobilized biomass and increased with hydraulic retention time reaching a maximum value of 1.6 mg ATP/g VS at hydraulic retention time 4.35 h. The ATP content of anaerobes in the UASB effluent declined rapidly under aerobic conditions following a 2nd-order kinetic model.

Evaluation of renal damage using urinary ATP analysis

It is reported that urinary ATP concentration analysis is useful for determining urinary tract infection and renal damage caused by drugs. By means of the firefly luciferin-luciferase method, we determined the reference value of urinary free ATP and evaluated the effects of urine sediments and conditions of storage. The reference value was established as 1.77 x 10(-10) to approximately 7.70 x 10(-9)M using urine samples obtained from 63 outpatients who seemed to have no renal disease. There was no significant difference in ATP concentration between 33 males and 30 females. No significant changes were observed in 11 healthy volunteers during a 1-year period. Within-run reproducibility of ATP was satisfying (8.28% and 11.4% of coefficient value in low and high concentration samples, respectively). ATP concentration was significantly decreased after centrifugation (p < 0.05) and after filtration (p < 0.01). The amounts of the red blood cells (RBC) and white blood cells (WBC) in samples whose ATP concentration was decreased after centrifugation or filtration were significantly higher than those in samples whose concentration did not decrease (p < 0.05). Urine containing many RBCs and/or WBCs might show an artificially higher ATP concentration if no preparations has been performed. There were significant positive correlations between the ATP concentrations before and after refrigeration, but no correlations before and after freezing. It is concluded that the reference value of urinary free ATP concentration was 1.77 x 10(-10) to approximately 7.70 x 10(-9) M and that care is required in the estimation of urinary ATP concentrations in samples containing many sediments, especially with WBC and RBC.

Principles and problems in physical chemistry for biochemists (3rd ed.)

Principles and problems in physical chemistry for biochemists (3rd ed.)

Glutathione Synthesis during the Rewarming of Rat Hepatocytes Preserved in the University of Wisconsin Solution

In this study, we used isolated rat hepatocytes to investigate the effect of nucleoside content of the preserved cells on the ability to synthesize glutathione (GSH) during the rewarming process. We cold-stored hepatocytes in University of Wisconsin (UW) solution (72 h, 0°C, N 2) without nucleosides and with the addition of 5 mM adenosine or 10 mM ATP. After 72 h of cold storage, we determined the GSH synthesis rate and the ATP content of the cells. We found a GSH synthesis rate similar to that of freshly isolated hepatocytes only in the group of cells cold-stored with 10 mM ATP. When we tested the cellular ATP concentrations, we found that controls and preserved cells with 10 mM ATP showed a similar value of ATP during the rewarming step. Our results suggested that the incorporation of ATP in the UW solution increased the ATP content and the rate of GSH synthesis of cold-stored hepatocytes during rewarming.

A re-evaluation of the ATP :NADPH budget during C3 photosynthesis: a contribution from nitrate assimilation and its associated respiratory activity?

The purpose of this review in reanalysing the ATP :reluctant balance in illuminated leaf cells is to stress that photosynthesis in vivo does not involve CO2 fixation alone, but embraces other processes, chief among which is N assimilation. Prior to the demonstration of CO2 fixation and photophosphorylation by isolated chloroplasts, the mitochondria were thought likely to provide all the ATP required for CO2 fixation (discussed in Arnon et al., 1954). During the 1960s, the development of techniques for the isolation of chloroplasts able to fix CO2 at rates approaching those of the parent tissue induced a paradigm shift, leading to the establishment of a dominant (if not unanimous) view that chloroplasts in vivo must by themselves meet all their ATP requirements. More recent studies, however, indicate that the reality lies somewhere between these two extremes. The present work places emphasis on the integrated nature of photosynthesis and proposes that much of the respiratory ATP necessary for whole cell photosynthesis may be generated during the production of C skeletons for N assimilation. Rather than considering dissipative electron transport pathways as necessary to uncouple respiratory precursor synthesis from ATP production, the present analysis emphasizes the metabolic value of ATP produced during N-linked respiration, with cellular ATP supply being tailored to ATP demand.

Effects of YT-146 [2-(1-Octynyl) Adenosine], an Adenosine A2A Receptot Agonist, on cAMP Production and Noradrenaline Release in PC12 Cells

Effects of YT-146 [2-(1-octynyl) adenosine], an adenosine A2 receptor agonist, on cAMP production and noradrenaline (NA) release were investigated in PC12 cells. YT-146 caused a concentration-dependent cAMP accumulation (EC50: 1.2±0.9 nM). In [3H]NA-prelabeled cells, YT-146 increased the basal NA release and enhanced ATP-evoked NA release in a concentration-dependent manner (EC50: 0.23±0.15 nM). YT-146 augmented the maximal response to ATP without affecting the EC50 value of ATP. These effects of YT-146 were inhibited by several adenosine receptor antagonists with a characteristic of adenosine A2A receptor subtype. The effects of YT-146 were mimicked by forskolin, dibutylyl cAMP and Sp-cAMPS, and inhibited by H-89, a cAMP-dependent protein kinase inhibitor. YT-146 had little effect on ATP-induced increase in intracellular Ca2+ concentration. YT-146 enhanced the NA release induced by several different stimuli including Ca2+ ionophore A23187. The present results suggest that YT-146 is a potent agonist on adenosine A2A receptors in PC12 cells and causes a cAMP-dependent enhancement of NA release by affecting the exocytosis process at a point downstream of the intracellular Ca2+ increase.