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Active nematics represent a relatively new class of materials with liquid crystalline properties that rely on an energy source to maintain their structure and dynamics. In this paper, we focus on the well-known microtubule-kinesin-based active nematic, powered by chemical energy as ATP (Adenosine triphosphate), and report on how ATP depletion impacts active nematic defect dynamics and texture. Using fluorescence microscope video imaging, we measure the time averaged mean defect separations and root mean square velocities of the active flows as a function of time. We also characterise textural changes and the growth of void space in the network after ATP depletion using an image binarization technique. [ABSTRACT FROM AUTHOR]

We applied a modified assay for the measurement of the intracellular ATP content as an alternative biochemical method for estimating the viability of the BCG vaccine instead of the standard cultural microbiological assay. Eight lyophilized BCG ampoule samples (Lot 287-2 and Lot 254-2, Bulgarian BCG strain SL222 Sofia) were tested on different days. Each calibration curve was measured for the following range of ATP concentrations: 100, 10, 5, 2.5, 1.25, 0.625, 0.3125, and 0.1562 pmol. The obtained results were compared to the traditional cultural colony counting method for evaluating the viability of the BCG vaccine. The NCP of Lot 287-2 (min 2.325–max 3.300) was higher in comparison to Lot 254-2 (min 1.438–max 3.330). That result corresponded to higher ATP-content registered for Lot 287-2 (min 0.409–max 1.338) than Lot 254-2 (min 0.18–max 1.104). Our results confirmed the observation made by other researchers that the modified ATP assay is a reliable, simple, and quick method for the evaluation of the viability of the BCG vaccine. Additional compiled data about all BCG strains are needed to perform studies on the correlation between the luciferin-luciferase bioluminescent ATP assay and the cultural method for viability and enumeration of the units in the lyophilized BCG vaccine. [ABSTRACT FROM AUTHOR]

Some G-quadruplex-hemin complexes can be used as a type of DNAzymes with peroxidase activity, it can efficiently catalyze H2O2-mediated oxidation reactions. Herein, a simple, colorimetric, and homogenous sensor design strategy based on G-quadruplex-hemin DNAzymes was developed. This strategy takes advantages of the competition between a DNA/DNA duplex and a DNA/target complex, and the formation of catalytically active G-quadruplexes in the presence of targets. With this method, no labeled oligonucleotides are needed, and no expensive instruments are required. This strategy may be developed as a general approach for the analysis of many metal ions and aptamer substrates. In this paper, the feasibility of this strategy in Hg2+ and adenosine triphosphate (ATP) detection was studied, and results showed that this “turn-on” process enabled the selective detection of aqueous Hg2+ and adenosine triphosphate (ATP) using a simple colorimetric technique. [ABSTRACT FROM AUTHOR]

Adenosine Triphosphate is a molecule with the function of generating energy, where high levels are directly related to cellular and muscular health. Oral supplementation appears as a possibility to guarantee such levels, and is normally consumed in capsules, which can be acute or chronic. To verify the influence of a dose of ATP on muscular performance in adults. This is an acute, crossover, randomized, double-blind, placebo-controlled intervention study. 18 trained men were recruited, with an average age of 27.95 years. Two visits were made to the laboratory, where each of the protocols consisted of randomized supplementation of 400 mg of Peak ATP® or placebo, and 30 min later, five series were performed, where the first consisted of five repetitions and the following four of ten repetitions at 60° per second in knee extension and flexion on a Biodex® 4.0 isokinetic dynamometer, where the volunteers' muscle strength and resistance to fatigue were measured. For data analysis, data normality was assessed using Shapiro Wilk, the ANOVA repeated measures test with Bonferroni post hoc. To identify the size of the effect, the Cohen test was performed, and the statistical package used was SPSS 25 with an applied significance of p < 0.05. Acute supplementation was unable to achieve significant improvements in muscle strength indicators. The supplement delays drops in strength as the exercise progresses in knee extension compared to the placebo. An acute dose of 400 mg of ATP did not improve the volunteers' muscle strength indicators, but it was able to reduce fatigue levels as the exercise progressed, enabling greater performance for longer. [ABSTRACT FROM AUTHOR]

Since the first reported observations by Zobell in 1943, it has been recognized that the metabolic activity of adhered bacteria can differ from that of their planktonic counterparts. Many studies have been performed and the overwhelming evidence is that bacterial adhesion to surfaces can result in changes to cellular metabolic activity and that the changes are a function of the surface properties of both the bacterial and adhering surfaces. However, the mechanism that results in these observations has remained elusive. The authors have approached this problem by focusing on cellular bioenergetics, which describes how bacteria obtain, store and use energy, and how adhesion can affect this process. In a series of experimental and numerical studies, the authors developed a hypothesis linking cellular bioenergetics to the physicochemical charge-regulation effect, which causes variations in surface electrostatic properties as a surface containing acid/base functional groups (e.g., the bacterial cell surface) approaches another surface. The purpose of this paper is to synthesize these prior studies and provide a cohesive presentation of the hypothesis. If this hypothesis is ultimately shown to be true, it will provide a fundamental basis for the engineered design of surface materials and coatings that can enhance or inhibit bacterial activity and colonization, depending on the requirements of the system at hand. [ABSTRACT FROM AUTHOR]

Aim: Cell viability assays are critical for cell-based products. Here, we demonstrate a combined experimental and computational approach to identify fit-for-purpose cell assays that can predict changes in cell proliferation, a critical biological response in cell expansion. Materials & methods: Jurkat cells were systematically injured using heat (45 ± 1°C). Cell viability was measured at 0 h and 24 h after treatment using assays for membrane integrity, metabolic function and apoptosis. Proliferation kinetics for longer term cultures were modeled using the Gompertz distribution to establish predictive models between cell viability results and proliferation. Results & conclusion: We demonstrate an approach for ranking these assays as predictors of cell proliferation and for setting cell viability specifications when a particular proliferation response is required. In recent years, there has been a surge in the amount of cellular therapy products which have been engineered to treat patients with severe diseases. These cellular products use living cells to treat the disease, and the quality of these cell products is critical for ensuring product safety and effectiveness. Throughout the process of engineering and manufacturing these cell products, many cells can die or be in the process of dying, and the amount of dead cells in the product can impact product yield and quality. In any given cell product at any given time during the manufacturing process, cells are exposed to stresses, and these stresses can injure the cells through several mechanisms, leading to a range of cell death events that can follow different timelines. There are many existing assays which evaluate the health of the cells, known as cell viability assays, and these assays can be based on many different cell features that indicate a cell has been injured (i.e., cell membrane permeability, changes in cell metabolism, molecular markers for cell death). These cell viability assays provide different insights into the state of cell health/injury based on what cell features are being evaluated and the timing at which the viability measurements are taken, and some viability assays may be more appropriate than others for specific applications. Therefore, a method is needed to appropriately select cell viability assays that are designed to evaluate injuries to cells that occur in specific bioprocess. In this series of studies, we used a range of analytical methods to study the number of living and dead cells in a series of cell populations that we treated to induce damage to the cells, reducing their ability to grow. We then used mathematical models to determine the relationship between cell viability measurements and cell growth over time, and used the results to determine the sensitivity of the viability assays to changes in cell growth. We used a specific cell line in this example, but this technique can be applied to any cell line or cell sample population and different types of injuries can be applied to the cells. This approach can be used by manufacturers of cell-based products and therapies to identify cell viability assays that are meaningful for monitoring the production of cells and characterizing product quality. [ABSTRACT FROM AUTHOR]

Ribociclib is a CDK4/6 inhibitor approved for the treatment of breast cancer; it inhibits the activity of CDK4/6 by competitively binding to adenosine 5'-triphosphate (ATP) binding sites. Although generally well-tolerated, ribociclib has been connected to a number of serious dermatologic complications. This study explored the effects of ATP on ribociclib-induced skin damage. Using a rat model, ATP 25 mg/kg was injected intraperitoneally in the ATP + Ribociclib (ATR) group (n = 6). Distilled water as solvent was applied to the healthy control (HC) group (n = 6) and ribociclib (RCB) group (n = 6). One hour after ATP and solvent administration, ribociclib (200 mg/kg) suspension prepared in distilled water was administered to the stomach by gavage (ATR and RCB groups). This was repeated once a day for 15 d. After that period, biochemical markers were studied in the skin tissues and histopathological evaluations were conducted. In the histopathological evaluation of the RCB group, dermal necrosis, degeneration in hair follicles, and pycnosis in keratinocytes were observed. Only mild degeneration was observed in the ATR group; the HC group had a normal histological appearance. The malondialdehyde (MDA) values were significantly higher and the superoxide dismutase (SOD), catalase (CAT), and total glutathione (tGSH) levels were significantly lower in the RCB group in comparison to the HC group (p <.001). ATP reduced the ribociclib-induced increases in the MDA values and decreased the SOD, CAT, and tGSH levels in the ATR group (p <.001). ATP may be useful in the treatment of ribociclib-induced skin damage. [ABSTRACT FROM AUTHOR]

Cancer and infectious diseases are among the leading causes of death in the world. Despite the diverse array of treatments available, challenges posed by resistance, side effects, high costs, and inaccessibility persist. In the Solanaceae plant family, few studies with Vassobia breviflora species relating to biological activity are known, but promising results have emerged. The phytochemicals present in the ethyl acetate fraction were obtained using ESI-MS-QTOF, and the antioxidants assays 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical capture (ABTS), plasma ferric reduction capacity (FRAP), and total antioxidant capacity (TAC). Cytotoxic activity was evaluated by MTT, Neutral Red, and lactate dehydrogenase (LDH) released. The production of reactive oxygen species, nitric oxide, and purinergic enzymes was also investigated. Antibacterial activity was measured through minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and antibiofilm activity, in addition to genotoxicity in plasmid DNA. Five major masses were identified D-glucopyranose II, allyl disulfide, γ-lactones, pharbilignoside, and one mass was not identified. V. breviflora exhibited relevant antioxidant and cytotoxic activity against the HeLa cell line and enhanced expression effect in modulation of purinergic signaling. Antibacterial activities in the assays in 7 ATCC strains and 8 multidrug-resistant clinical isolates were found. V. breviflora blocked biofilm formation in producing bacteria at the highest concentrations tested. However, there was no plasmid DNA cleavage at the concentrations tested. Data demonstrated that V. breviflora exhibited an antioxidant effect through several methods and proved to be a promising therapeutic alternative for use against tumor cells via purinergic signaling and multidrug-resistant microorganisms, presenting an anti-biofilm effect. [ABSTRACT FROM AUTHOR]

Nucleotides and platelets have been associated with a wide range of activities that affect the host inflammatory response. The main goal of this study is to examine the roles of nucleotide signaling and platelets in inflammation. The study analysis entailed conducting a systematic search to identify relevant articles in PsycINFO, PubMed, Web of Science, and CINAHL. The evidence gathered from the identified articles shows the roles of nucleotides and platelets in inflammation. In the extracellular environment, nucleotides act as signaling molecules that can activate nucleotide receptors to promote inflammation. Inflammation is an essential process through which the innate immune system responds to pathogens, microbes, and damage-associated molecular patterns. Moreover, research evidence shows that the mechanisms through which platelets affect inflammatory responses and regulate hemostasis are the same. The roles of nucleotides and platelets in inflammation have been explored in several studies worldwide. Although platelets and nucleotides have unique structures, both of them influence the host response to pathogens and tumors. Analysis of platelets and nucleotides will offer valuable insight for the development of new treatments for infectious and inflammatory diseases. 1. Nucleotide triphosphates, such as ATP, have a secret "second life" in addition to being the most important energy-storing molecule inside cells. Outside of cells and in the bloodstream, they are important signaling molecules in the context of inflammation. 2. As platelets have nucleotide receptors on their surfaces, there is important cross-talk between platelet activation and nucleotide signaling in the context of inflammation. [ABSTRACT FROM AUTHOR]

ATP metabolism during mitosis needs to be coordinated with numerous energy-demanding activities, especially in cancer cells whose metabolic pathways are reprogramed to sustain rapid proliferation in a nutrient-deficient environment. Although strategies targeting the energy metabolic pathways have shown therapeutic efficacy in preclinical cancer models, how normal cells and cancer cells differentially respond to energy shortage is unclear. In this study, using time-lapse microscopy, we found that cancer cells displayed unique mitotic phenotypes in a dose-dependent manner upon decreasing ATP (i.e. energy) supply. When reduction in ATP concentration was moderate, chromosome movements in mitosis were barely affected, while the metaphase–anaphase transition was significantly prolonged due to reduced tension between the sister-kinetochores, which delayed the satisfaction of the spindle assembly checkpoint. Further reduction in ATP concentration led to a decreased level of Aurora-B at the centromere, resulting in increased chromosome mis-segregation after metaphase delay. In contrast to cancer cells, ATP restriction in non-transformed cells induced cell cycle arrest in interphase, rather than causing mitotic defects. In addition, data mining of cancer patient database showed a correlation between signatures of energy production and chromosomal instability possibly resulted from mitotic defects. Together, these results reveal that energy restriction induces differential responses in normal and cancer cells, with chromosome mis-segregation only observed in cancer cells. This points to targeting energy metabolism as a potentially cancer-selective therapeutic strategy. [ABSTRACT FROM AUTHOR]

ATPases and GTPases are two important classes of protein that play critical roles in energy transduction, cellular signalling, gene regulation and catalysis. These proteins use cofactors such as nucleoside di and tri-phosphates (NTP, NDP) and can detect the difference between NDP and NTP which then induce different protein conformations. Mechanisms that drive proteins into the NTP or NDP conformation may depend on factors such as ligand structure and how Mg2+ coordinates with the ligand, amino acids in the pocket and water molecules. Here, we have used the advanced electrostatic and polarisable force field AMOEBA and molecular dynamics free energy simulations (MDFE) to examine the various binding mechanisms of ATP:Mg2+ and ADP:Mg2+. We compared the ATP:Mg2+ binding with previous studies using non-polarisable force fields and experimental data on the binding affinity. It was found that the total free energy of binding for ATP:Mg2+ (−7.00 ± 2.13 kcal/mol) is in good agreement with experimental values (−8.6 ± .2 kcal/mol) [1]. In addition, parameters for relevant protonation states of ATP, ADP, GTP and GDP have been derived. These parameters will allow for researchers to investigate biochemical phenomena involving NTP's and NDP's with greater accuracy than previous studies involving non-polarisable force fields. [ABSTRACT FROM AUTHOR]

Cordyceps militaris has been reported to the diverse pharmaceutical effects including cancer, inflammatory diseases, and bacteria or virus infection. However, the effect of C. militaris on exercise performance has not yet been elucidated. In this study, we investigated the beneficial effect of C. militaris on exercise performance. To evaluate exercise performance, we prepared C. militaris ethyl acetate extract (CMEE) and conducted grip strength tests every week after administration. Additionally, blood samples were collected at the end of the experiment for biochemical analysis. The administration of CMEE slightly increased grip strength, and this result was similar to the red ginseng treated group. According to the result of biochemical analysis, CMEE had an effect on the biomarkers related to ATP generation pathway but had little influence on the muscle fatigue related biomarkers. Therefore, C. militaris has the possibility of improving exercise performance, which could be associated with the increase in ATP production rather than the decrease in muscle fatigue during exercise. [ABSTRACT FROM AUTHOR]

Objectives. Heart failure (HF) impairs resting myocardial energetics, myocardial mitochondrial performance, and maximal oxygen uptake (VO2max). Exercise training is included in most rehabilitation programs and benefits HF patients. However, the effect of exercise intensity on cardiac mitochondrial respiration and concentrations of the key bioenergetic metabolites phosphocreatine (PCr), adenosine triphosphate (ATP), and inorganic phosphate (Pi) is unclear. This study aimed to investigate the effects of exercise training at different intensities in rats with HF. Methods. Rats underwent myocardial infarction or sham operations and were divided into three subgroups: sedentary, moderate intensity, or high intensity. The impact of HF and 6 weeks of exercise training on energy metabolism was evaluated by 31P magnetic resonance spectroscopy and mitochondrial respirometry. The concentrations of PCr, ATP, and Pi were quantified by magnetic resonance spectroscopy. VO2max was measured by treadmill respirometry. Results. Exercise training increased VO2max in sham and HF. PCr/ATP ratio was reduced in HF (p < .01) and remained unchanged by exercise training. PCr concentration was significantly lower in HF compared to sham (p < .01). Moderate and high-intensity exercise training increased ATP in HF and sham. HF impaired complex I (CI) and complex II (p = .034) respiration. High-intensity exercise training recovered CI respiration in HF rats compared to HF sedentary (p = .014). Conclusions. Exercise training improved cardiac performance, as indicated by increased VO2max and higher exercise capacity, without changing the myocardial PCr/ATP ratio. These observations suggest that the PCr/ATP biomarker is not suited to evaluate the beneficial effects of exercise training in the heart. The exact mechanisms require further investigations, as exercise training did increase ATP levels and CI respiration. [ABSTRACT FROM AUTHOR]

Acetyl phosphate (AcP) is generally produced from acetyl coenzyme A by phosphotransacetylase (Pta), and subsequent reaction with ADP, catalyzed by acetate kinase (Ack), produces ATP. The mechanism of ATP production in Porphyromonas gingivalis is poorly understood. The aim of this study was to explore the molecular basis of the Pta-Ack pathway in this microorganism. Pta and Ack from P. gingivalis ATCC 33277 were enzymatically and structurally characterized. Structural and mutational analyses suggest that Pta is a dimer with two substrate-binding sites in each subunit. Ack is also dimeric, with a catalytic cleft in each subunit, and structural analysis indicates a dramatic domain motion that opens and closes the cleft during catalysis. ATP formation by Ack proceeds via a sequential mechanism. Reverse transcription-PCR analysis demonstrated that the pta (PGN_1179) and ack (PGN_1178) genes, tandemly located in the genome, are cotranscribed as an operon. Inactivation of pta or ack in P. gingivalis by homologous recombination was successful only when the inactivated gene was expressed in trans. Therefore, both pta and ack genes are essential for this microorganism. Insights into the Pta-Ack pathway reported herein would be helpful to understand the energy acquisition in P. gingivalis. [ABSTRACT FROM AUTHOR]

1. Guanidinoacetic acid (GAA) is the single endogenous precursor of creatine, which plays a critical role in energy homeostasis of cells. Since GAA is endogenously converted to creatine by methylation, it was hypothesised that the effects of dietary GAA supplementation might determine the methionine (Met) availability in corn-soybean based diets. 2. A total of 540, one-day-old male Ross 308 broilers were allocated to nine dietary treatments with six replicates (10 birds each) in a 3 × 3 factorial arrangement with three graded levels of supplementary Met (+0.4 g/kg per level), whilst cystine was equal across groups, resulting in a low, medium and high level of total sulphur amino acids, and with three levels of GAA (0, 0.6 and 1.2 g/kg). Birds were fed for 42 days. 3. Increasing levels of supplemental Met enhanced performance indices in all rearing periods, although there was no effect on feed conversion ratio in the grower or feed intake in the finisher periods. Final body weight was 8.8% and 14.6% higher in the birds fed medium and high Met diets, respectively, compared to the low Met level. Relative breast weight and protein content in muscle on d 25 linearly increased with higher levels of Met. At low and high Met levels, growth in the finisher phase was negatively affected by supplementing GAA at 1.2 g/kg. It was suggested that disturbances in methylation homeostasis and/or changes in Arg metabolism might explain these findings. At the end of the grower phase, muscle creatine content was higher when feeding GAA at 0.6 and 1.2 g/kg (4464 and 4472, respectively, vs. 4054 mg/kg fresh muscle in the control group). 4. The effects of dietary GAA supplementation were influenced by the dietary Met level only in the finisher period, which indicates the need for proper sulphur amino acid formulation in diets when feeding GAA. [ABSTRACT FROM AUTHOR]

(1) Guanidinoacetic acid (GAA) is the single immediate endogenous precursor of creatine (Cr). It was hypothesised that dietary GAA would have different effects on performance and energy metabolites in breast muscle depending on the nutrient density (ND) of corn-soybean-based diets. (2) A total of 540 one-day-old male Ross 308 broilers were allocated to 9 dietary treatments with 6 replicates (10 birds each) in a 3 × 3 factorial arrangement with three levels of ND (low, 2800; medium, 2950 and high, 3100 kcal metabolizable energy (ME)/kg; and with the other nutrients being constant relative to ME) and supplemented with three levels of GAA (0, 0.6 and 1.2 g/kg) in a 42-d feeding trial. (3) In the starter and grower periods, increasing levels of ND improved body weight (BW), average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR), with the exception of ADFI in the starter period. GAA supplementation did not affect performance characteristics. All performance indicators responded markedly to increasing ND in the finisher period, whereas the highest GAA level reduced ADFI compared to the unsupplemented control (156 vs. 162 g/d) and concomitantly FCR (1.81 vs. 1.93). No interactive effects were noted for any performance trait. The high ND diet resulted in more breast meat yield on d42, associated with higher fat content and darker colour compared to the other ND levels. The GAA supplementation did not affect carcass and breast traits. At the end of the experiment, Cr was elevated when feeding GAA at 1.2 g/kg (5455 vs. 4338 mg/kg fresh muscle). (4) To conclude, ND had a substantial effect on performance and carcass traits, whereas any effect of GAA was limited to FCR in the finisher period and independent of diet ND level. [ABSTRACT FROM AUTHOR]

Introduction: The adenosine pathway plays a key role in modulating immune responses in health and in disease. In health, anti-inflammatory effects of adenosine balance pro-inflammatory ATP, limiting tissue destruction by activated immune cells. In disease, this balance is disturbed. Areas covered: This review focuses on cancer and explains how in the microenvironment, the ATP-adenosine balance shifts towards an excess of extracellular adenosine Expert commentary: The CD73-adenosine axis plays a key role in the inhibition of anti-tumor functions of immune effector cells. Today, adenosine emerges as one of the immune checkpoints that are implicated in the tumor escape from the host immune system. The adenosine pathway is currently viewed as a significant barrier to the effectiveness of immune therapies and becomes an important therapeutic target in cancer. Pharmacologic inhibitors or antibodies specific for the components of the adenosine pathways or adenosine receptors show efficacy in pre-clinical studies and are entering the clinical arena. [ABSTRACT FROM AUTHOR]

In this study we looked at smokers with and without chronic obstructive pulmonary disease (COPD) patients in order to evaluate the incidence of 4977 base pair (bp) mtDNA (mtDNA4977) deletion and mtDNA copy number in sputum cells and in peripheral blood leukocytes (PBLs) in relation to mitochondrial function and oxidative stress status. Twenty-five COPD patients who were current smokers, 22 smokers and 23 healthy nonsmokers (for only PBLs studies) participated in this study. The 4977-bp deletion was detected in all examined samples within 40 cyles of PCR amplification, using a quantitative real time PCR. The frequency of the mtDNA4977 was significantly higher in the sputum cells of patients with COPD compared to smokers without COPD (p < 0.0001). This difference was not observed in PBLs. Levels of cellular oxidative stress were significantly higher in the sputum cells of subjects with COPD than in the smoker group. However, mtDNA copy number, mitochondrial membrane potential (ΔΨm) and cellular ATP levels in PBLs and sputum cells were not significantly different between the studied groups. The Pearson analysis revealed no correlations between the accumulation of mtDNA4977, and intracellular ATP content and ΔΨm values of the sputum cells, although there was a positive correlation between the increase in the percentage of deleted mtDNA4977 and the levels of cellular oxidative stress in COPD patients (r = 0.80, p < 0.0001). Our studies may suggest that the accumulation of mtDNA4977 in the sputum cells of smokers with COPD does not seem to have an important impact on mitochondrial dysfunction in relation to ATP production and ΔΨm when compared to those of healthy smokers. [ABSTRACT FROM AUTHOR]

Purpose: To analyze the dose rate influence in hyper-radiosensitivity (HRS) of human melanoma cells to very low doses of fast neutrons and to compare to the behaviour of normal human skin fibroblasts. Materials and methods: We explored different neutron dose rates as well as possible implication of DNA double-strand breaks (DSB), apoptosis, and energy-provider adenosine-triphosphate (ATP) levels during HRS. Results: HRS in melanoma cells appears only at a very low dose rate (VLDR), while a high dose rate (HDR) induces an initial cell-radioresistance (ICRR). HRS does not seem to be due either to DSB or to apoptosis. Both phenomena (HRS and ICRR) appear to be related to ATP availability for triggering cell repair. Fibroblast survival after neutron irradiation is also dose rate-dependent but without HRS. Conclusions: Melanoma cells or fibroblasts exert their own survival behaviour at very low doses of neutrons, suggesting that in some cases there is a differential between cancer and normal cells radiation responses. Only the survival of fibroblasts at HDR fits the linear no-threshold model. This new insight into human cell responses to very low doses of neutrons, concerns natural radiations, surroundings of accelerators, proton-therapy devices, flights at high altitude. Furthermore, ATP inhibitors could increase HRS during high-linear energy transfer (high-LET) irradiation. [ABSTRACT FROM AUTHOR]

In this study the effect of volatile to total solids (VS/TS) on anaerobic digestion of waste activated sludge (WAS) pretreated by alkaline, thermal and thermal–alkaline strategies was studied. Experimental results showed that the production of methane from sludge was increased with VS/TS. When anaerobic digesters were fed with sludge pretreated by the thermal–alkaline method, the average methane yield was improved from 2.8 L/d at VS/TS 0.35 to 4.7 L/d at VS/TS 0.56. Also, the efficiency of VS reduction during sludge anaerobic digestion varied between 18.9% and 45.6%, and increased gradually with VS/TS. Mechanism investigation of VS/TS on WAS anaerobic digestion suggested that the general activities of anaerobic microorganisms, activities of key enzymes related to sludge hydrolysis, acidification and methanogenesis, and the ratio ofArchaeatoBacteriawere all increased with VS/TS, showing good agreement with methane production. [ABSTRACT FROM AUTHOR]

Purpose: We determined nitric oxide (NO) productionviainducible NO synthase (iNOS) by hyperglycemia using the retina of Otsuka Long-Evans Tokushima Fatty rats (OLETF rats), and investigated the relationship between ATP contents and NO production in the retinas of OLETF rats. Methods: Long-Evans Tokushima Otsuka rats (LETO rats, normal rats) and OLETF rats (model rat for diabetes mellitus) aged 60 weeks of age were used. Plasma glucose (Glu) levels were determined using an Accutrend GCT System, and NO levels were measured by the microdialysis method as nitrite (). Cytochromecoxidase (CCO) activity was measured using a Mitochondrial Isolation Kit and CytochromecOxidase Assay Kit, and ATP levels were determined using a Sigma ATP Bioluminescent Assay Kit and a luminometer AB-2200. Results:levels in the retinas of OLETF rats were significantly higher than in LETO rats, and thelevels in the retinas of 60-week-old OLETF rats increased with increasing Glu. CCO activity in the retinas of OLETF rats showed no significant difference from that in LETO rats; however, ATP levels in the retinas of OLETF rats were significantly lower than those in LETO rats. The oral administration of aminoguanidine or disulfiram, an iNOS inhibitor, attenuated the decrease in ATP levels in the retinas of 60-week-old OELTF rats. Conclusion: The present study demonstrates that NO productionviaiNOS in the retinas of 60-week-old OLETF rats is caused by hyperglycemia, and that NO causes a decrease in ATP contents in the retinas of 60-week-old OELTF rats. It is possible that the low ATP contents caused by NO may affect the normal functioning of the retina in OLETF rats. [ABSTRACT FROM PUBLISHER]

Since the first implant in 1980, implantable cardioverter defibrillator (ICD) technology has progressed rapidly. Modern ICD’s have hundreds of programmable options with the general goal of preventing inappropriate shocks and providing shocks for truly life threatening symptomatic ventricular arrhythmias. New studies on ICD programming have shown the benefits of prolonged detection intervals in reaching this goal. Anti-tachycardia pacing (ATP) therapy has become an important adjunct to defibrillator shocks. Remote monitoring technologies have surfaced which have been shown to identify arrhythmias and problems with the device in an expedient fashion. The subcutaneous ICD offers the advantage of avoiding intravascular leads and their inherent risks. Lastly, the current understanding of the effects of MRI in ICD patients has advanced creating new opportunities to provide MRI safely to such patients. [ABSTRACT FROM PUBLISHER]

Autophagic degradation of proteasomes (termed proteaphagy) is a conserved mechanism by which cells eliminate excess or damaged particles. This clearance is induced rapidly when organisms are starved for nitrogen and, because proteasomes are highly abundant, their breakdown likely makes an important contribution to the amino acid pools necessary for survival. By contrast, our recent studies found that proteasomes are not degraded in response to carbon starvation, even though bulk macroautophagy is similarly activated. Instead, carbon starvation induces sequestration of proteasomes into membrane-less cytoplasmic condensates previously termed proteasome storage granules (PSGs), which protect proteasomes from autophagic degradation. Preserving proteasomes in PSGs enhances the ability of yeast cells to recover from a variety of stresses, implying that rapid remobilization of stored proteasomes when conditions improve is advantageous to cell fitness. Consequently, the choice of whether to save or degrade proteasomes can profoundly impact cell survival. [ABSTRACT FROM AUTHOR]

To test the hypothesis that a fall in cellular ATP following stimulation of endothelial cells with thrombin is secondary to a decrease in NAD levels caused by poly(ADP-Ribose)polymerase (PARP), we measured the levels of NAD and ATP in endothelial cells after treatment with thrombin, the Ca++-ionophore A23187, or hydrogen peroxide (H2O2), and compared the effects of inhibitors of PARP, NAD synthesis, and ADP-ribose breakdown on these responses. Neither thrombin nor A23187 caused a reduction in endothelial NAD levels and A23187 affected ATP levels independently of NAD levels or PARP activity. H2O2induced lowering of NAD caused modest lowering of ATP but marked additional ATP-lowering, independent of PARP and NAD, was also demonstrated. We conclude that in endothelial cells ATP levels are largely independent of NAD and PARP, which do not play a role in thrombin or Ca++-ionophore-mediated lowering of ATP. H2O2caused ATP lowering through a similar mechanism as thrombin and A23187 but, additionally, caused a further ATP lowering through its intense stimulation of PARP and marked lowering of NAD. [ABSTRACT FROM PUBLISHER]

Previous studies have suggested that carbon monoxide (CO) poisoning stimulates cAMP production via purine P2Y11-like receptors in the rat striatum, activating cAMP signaling pathways, resulting in hydroxyl radical (•OH) production. Extracellular ATP was thought likely to trigger the cascade, but the present study has failed to demonstrate a clear increase in the extracellular ATP due to CO poisoning. The CO-induced •OH production was attenuated by the P2Y11 receptor antagonist NF157, in parallel with its abilities to suppress the CO-induced cAMP production. The •OH production was more strongly suppressed by a non-selective P2 receptor antagonist, PPADS, which had no effect on cAMP production. More selective antagonists toward the respective P2 receptors susceptible to PPADS, including NF279, had little or no effect on the CO-induced •OH production. The intrastriatal administration of exogenous ATP dose-dependently stimulated •OH production, which was dose-dependently antagonized by PPADS and NF279 but not by NF157. Exogenous GTP and CTP dose-dependently stimulated •OH production, though less potently. The GTP-induced •OH production was susceptible to both of NF279 and PPADS, but the CTP-induced •OH production was resistant to PPADS. The mechanism of •OH production may differ between CO poisoning and exogenous ATP, while multiple P2 receptors could participate in •OH production. The CO-induced •OH production was susceptible to the inhibition of NADPH oxidase, but not xanthine oxidase. Also, the NADPH oxidase inhibition suppressed •OH production induced by forskolin, a stimulator of intracellular cAMP formation. It is likely that •OH is produced by NADPH oxidase activation via cAMP signaling pathways during CO poisoning. [ABSTRACT FROM AUTHOR]

The adenosinergic pathway plays an important role in cancer progression. Aside from regulating functions of tumor cells and tissue cells present in the tumor microenvironment, extracellular adenosine is an autocrine or paracrine factor with powerful immunoregulatory activity. Adenosine signaling downregulates functions of most immune effector cells but enhances expansion and activity of immune cells responsible for suppression of anti-tumor immune responses. Adenosine is critical for limiting potential tissue-destructive effects of activated immune cells. It also facilitates tumor escape from the immune control. This review illustrates the involvement of adenosine and its four receptors, A1R, A2AR, A2BR and A3R, in the complex regulation of cellular and molecular cross talk that contributes to cancer progression. It also considers the potential of therapeutics targeting the adenosinergic pathway for benefiting cancer patients. [ABSTRACT FROM AUTHOR]

A novel cationic water-soluble polythiophene derivative, the poly[N,N,N-trimethyl-3-(thiophen-3-yl)propenaminium chloride] (PTPA-Cl) was chosen as a sensitive colorimetric and fluorescent probe to differentiate adenosine triphosphate (ATP), the present work shows for the first time the difference between ATP and coenzyme A (CoA). ATP has much stronger colorimetric and fluorescent than CoA. When bound to different charged analytes, the polythiophene derivative probe emitted light with different colors due to the unique spectral properties of the conformation sensitive probe, and the sensing interaction can be apparently observed by naked eyes. Moreover, the cationic water-soluble polythiophene shows selectivity to ATP in the presence of other charged analytes and as well a high sensitivity with the detection limit at 10−8 M by using fluorometry at aqueous solution. [ABSTRACT FROM PUBLISHER]

Context: Previous in vitro studies have demonstrated that emodin (1,3,8-trihydroxy-6-methyl-anthraquinone), an anthraquinone derivative from the rhizome of Rheum palmatum L., can inhibit the activation of P2X7 receptors (P2X7R) as a potential antagonist. However, the effects of emodin on P2X7R-related inflammatory processes remain unclear. Objective: This study aimed to investigate the effects of emodin on different inflammation responses of macrophages induced by ATP, the natural ligand of P2X7R. Materials and methods: Rat peritoneal macrophages were treated with millimolar ATP and emodin (0.1, 0.3, 1, 3, 10 µM) or brilliant blue G (BBG, 0.1, 1, 10 µM). Cytosolic Ca2+ concentration ([Ca2+]c) was detected by fluorescent Ca2+ imaging. Interleukin-1β (IL-1β) release was measured by rat IL-1β ELISA kits. Reactive oxygen species (ROS) generation was examined by dihydroethidium (DHE) fluorescent staining. Phagocytic activity was tested by neutral red uptake assay. Results: We found that the [Ca2+]c increase evoked by ATP (5 mM) was inhibited by emodin, in a dose-dependent manner with IC50 of 0.5 μM. Furthermore, emodin reduced the IL-1β release induced by ATP (2 mM) in lipopolysaccharide (LPS)-activated macrophages, with an IC50 of 1.6 μM. Emodin also strongly suppressed the ROS production and phagocytosis attenuation triggered by ATP (1 mM), with IC50 values of 1 μM and 0.7 μM, respectively. Besides, BBG, a specific antagonist of P2X7R, exhibited similar suppressive effects on these inflammation responses. Conclusion: These results showed the inhibitory effects of emodin on ATP-induced [Ca2+]c increase, IL-1β release, ROS production and phagocytosis attenuation in rat peritoneal macrophages, by inhibiting the activation of P2X7R. [ABSTRACT FROM AUTHOR]

Purpose: To detect the possible biochemical signs of inflammatory activation in the peripheral circulation in a rodent model of hippocampus irradiation, and to examine the effects of L-alpha-glycerylphosphorylcholine (GPC) in this experimental protocol. Materials and methods: Anesthetized Sprague-Dawley rats were subjected to 40 Gy cobalt irradiation of both hemispheres of the hippocampus, with or without GPC treatment (50 mg/kg intravenously (i.v.), 5 min before the irradiation, n = 6, each). A third group ( n = 6) served as saline-treated control. Blood samples were obtained 3 h after the end of irradiation in order to examine the changes in plasma histamine, tumor necrosis factor-alpha (TNF-α), interleukin 1-beta, interleukin 6 (IL-6) and interleukin 10 (IL-10); liver tissue samples were taken to determine adenosine triphosphate (ATP) concentrations. Results: The hepatic ATP levels were significantly declined, while plasma concentrations of circulating TNF-α, IL-6, IL-10 and histamine were significantly increased after hippocampus irradiation. GPC treatment significantly reduced the irradiation-induced release of cytokines and histamine, and the liver ATP level was maintained at the control value. Conclusions: Targeted brain irradiation produced measurable pro- and anti-inflammatory cytokine changes in the systemic circulation. GPC supplementation provides significant protection against irradiation-induced peripheral pro-inflammatory activation and ATP depletion. [ABSTRACT FROM AUTHOR]

Purpose: We previously found two mechanisms for the dysfunction in Ca2+ regulation caused by excessive nitric oxide (NO) using the lenses of hereditary cataract model rats: the first is that NO causes a decrease in Adenosine-5′-triphosphate (ATP) level via cytochrome c oxidase (CCO), resulting in a decrease in ATPase function; the second is that NO causes enhanced lipid peroxidation, resulting in the oxidative inhibition of Ca2+-ATPase. In this study, we demonstrate the effect of excessive NO on lipid peroxidation and ATP production in human lens using a human lens epithelial cell line, SRA 01/04 (human lens epithelial (HLE) cells). Methods: Excessive NO via inducible NO synthase (iNOS) was induced by stimulating cells with a combination of interferon-gamma (1000 IU IFN-γ) and lipopolysaccharide (100 ng/mL LPS). CCO activity was measured using a Mitochondrial Isolation kit and Cytochrome c Oxidase Assay kit, and ATP levels were determined using a Sigma ATP Bioluminescent Assay Kit and a luminometer AB-2200. Results: Cytochrome c oxidase activity and ATP levels were decreased in HLE cells stimulated with IFN-γ and LPS, and aminoguanidine (AG) and diethyldithiocarbamate (DDC) added 6 h before cell collection significantly attenuated these decreases in cells stimulated with the IFN-γ and LPS for 24-30 h. However, the lower CCO activity and ATP levels in HLE cells stimulated with the IFN-γ and LPS for 30 h were not changed by treatment with AG or DDC for 6-12 h, while the CCO activity and ATP levels in HLE cells treated with AG or DDC for 18 were recovered. Conclusion: Excessive NO causes a decrease in CCO activity and ATP levels, and the recovery time for CCO activity is related to exposure time to NO in HLE cells. [ABSTRACT FROM AUTHOR]

Objective. N-acetylcysteine (NAC) is an antioxidative molecule known to protect liver tissue from oxygen radical species generated during ischemia and reperfusion (IR). Nutritional and toxicology studies have shown that NAC also improves glucose metabolism and glycogen stores. We hypothesized that NAC improves glycogenesis and that impaired glycogenesis is a key element in IR injury. Material and Methods. In an experimental model, 80 min of segmental liver ischemia was induced in 16 pigs and the reperfusion was followed for 360 min. Eight animals received NAC 150 mg/kg as a bolus injection followed by an infusion of NAC 50 mg/kg/h intravenously. Results. AST and leukocyte density were lower in the NAC-treated animals, unrelated to the glutathione levels or apoptosis. Glycogen stores returned to a higher degree in the NAC-treated animals and microdialysis revealed lower levels of lactate during the reperfusion phase. Nitrite/Nitrate levels in the NAC group were lower in both serum and microdialysates, indicating that NAC scavenges radical nitrosative species. Conclusions. NAC treatment improves glycogenesis after liver IR injury and reduces the level of intraparenchymal lactate during reperfusion, possibly due to the scavenging of radical nitrosative species. [ABSTRACT FROM AUTHOR]

Abstract: We have previously reported that derivatization of hemoglobin with periodate-modified sugar derivatives such as oxidized adenosine triphosphate (oATP) leads to an increase in prooxidant reactivity at the heme. Here, we report that copolymerization of hemoglobin with serum albumin alleviates this problem completely, to the extent where the copolymer even has a slightly lower autooxidation rate compared to native hemoglobin. A similar, although not as potent, effect is obtained when hemoglobin is derivatized with oATP in the presence of small-molecule antioxidants instead of albumin. [ABSTRACT FROM AUTHOR]

Adenosine triphosphate (ATP) is released by alveolar epithelial cells during ventilator-induced lung injury (VILI) and regulates fluid transport across epithelia. High CO2 levels are observed in patients with 'permissive hypercapnia,' which inhibits alveolar fluid reabsorption (AFR) in alveolar epithelial cells. The authors set out to determine whether VILI affects AFR and whether the purinergic pathway is modulated in cells exposed to hypercapnia. Control group was compared against VILI (tidal volume [[Vt]] == 35 mL/kg, zero positive end-expiratory pressure [[PEEP]]) and protective ventilation (Vt == 6 mL/kg, PEEP == 10 cm H2O) groups. Lung mechanics, histology, and AFR were evaluated. Alveolar epithelial cells (AECs) were loaded with Fura 2-AM to measure intracellular calcium in the presence ATP (10 μM) at 5%% or 10%% CO2 as compared with baseline. High tidal volume ventilation impairs lung mechanics and AFR. Hypercapnia (HC) increases intracellular calcium levels in response to ATP stimulation. HC ++ ATP is the most detrimental combination decreasing AFR. Purinergic signaling in AECs is modulated by high CO2 levels via increased cytosolic calcium. The authors reason that this modulation may play a role in the impairment of alveolar epithelial functions induced by hypercapnia. [ABSTRACT FROM AUTHOR]

Objective. Extracellular inorganic pyrophosphate (ePPi) is a key regulator of pathologic mineralization in articular cartilage. Articular chondrocytes generate ePPi by the transportation of intracellular PPi (iPPi) through transport mechanisms such as ANK or by the degradation of extracellular adenosine triphosphate (eATP) by ectoenzymes. Although numerous modulators of ePPi have been characterized, little is known about eATP elaboration in cartilage. We sought to determine (1) whether eATP is coordinately regulated with ePPi and (2) whether ANK transports ATP. Methods. Primary articular chondrocytes were treated with factors known to modulate ePPi levels including growth factors (TGFββ1 and IGF-1), anion channel inhibitors, and chemicals that alter adenylyl cyclase and protein kinase C activities. Additional chondrocyte monolayers were infected with adenovirus containing functional (Ad-ANK) or mutated (Ad-ANK mutant) ANK sequences. eATP levels were measured with a bioluminescent assay. Results. TGFββ1 enhanced eATP accumulation by 33%%, whereas IGF-1 decreased eATP accumulation by 63%% and attenuated TGFββ1-induced eATP release by 72%%. Forskolin and probenecid diminished eATP accumulation by 55%% and 89%%. Phorbol-12-myristate-13-acetate increased eATP by 29%%. Transfection of chondrocytes with Ad-ANK caused a 10-fold increase in eATP compared with control values. Conclusion. Modulation of eATP by various factors paralleled their effects on ePPi production, suggesting a shared pathway of ePPi and eATP production and implicating ANK in eATP transport. As eATP directly contributes to pathologic mineralization in articular cartilage, understanding eATP regulation may lead to effective therapies for crystal-associated arthritis. [ABSTRACT FROM AUTHOR]

Phagocytosis of naturally dying cells usually blocks inflammatory reactions in host cells. We have recently observed that clearance of cells dying through autophagy leads to a pro-inflammatory response in human macrophages. Investigating this response further, we found that during engulfment of MCF-7 or 293T cells undergoing autophagic death, but not apoptotic or anoikic ones, caspase-1 was activated and IL-1β was processed, then secreted in an MyD88-independent manner. Autophagic dying cells were capable of preventing some LPS-induced pro-inflammatory responses, such as TNFα, IL-6 and IL-8 induction, but synergized with LPS for IL-1β production. Caspase-1 inhibition prevented macrophage IL-1β release triggered by the dying cells and also other pro-inflammatory cytokines which were not formed in the presence of IL-1 receptor antagonist anakinra either. IL-1β secretion was also observed using calreticulin knockdown or necrostatin-treated autophagic MCF-7 cells and it required phagocytosis of the dying cells which led to ATP secretion from macrophages. Blocking K+ efflux during phagocytosis, the presence of apyrase, adding an antagonist of the P2X7 receptor or silencing the nod-like receptor protein NALP3 inhibited IL-1β secretion. These data suggest that during phagocytosis of autophagic dying cells ATP, acting through its receptor, initiates K+ efflux, inflammasome activation and secretion of IL-1β, which initiates further pro-inflammatory events. Thus, autophagic death of malignant cells and their clearance may lead to immunogenic response. [ABSTRACT FROM AUTHOR]

Minimizing drug interactions is paramount to improving the efficacy and tolerability of cancer therapy. The zebrafish represents an innovative cancer model due to highly conserved genetics and inherent capacity for high-throughput chemical screening. This pilot study extends the utility of the zebrafish to a preclinical model for pharmacodynamics by examining the interaction of the nucleoside analogue, cladribine with the calcium channel blocker, diltiazem. Cladribine (0.7–3.5 mM) and/or diltiazem (2.4 mM), was injected intraperitoneally into adult zebrafish and red blood cell (RBC) lysates were assayed by HPLC for levels of purine nucleotides (e.g. ATP), potential biomarkers of cardiovascular health. Diltiazem increased RBC ATP concentrations, which were inhibited by co-injection of cladribine. These results suggest a novel drug interaction and highlight the feasibility of the zebrafish as an in vivo model for pharmacodynamic studies. [ABSTRACT FROM AUTHOR]

An original ligand (Lac-10-Chol) designed to interact with asialoglycoprotein receptors to potentially target hepatocyte was synthesised by grafting a lactose head to a cholesteryl structure, which was then included in liposomes. Preliminary formulation tests led to the selection of conventional formulations based on soybean phosphatidylcholine/cholesterol/DOTAP (± DOPE) (± Lac-10-Chol) that present reproducible absolute entrapment value (1.45 ± 0.10%), with a size of 109 ± 7 nm and a slight positive charge (3.77 ± 1.59 mV). Cell viability (via the MTT test), expressed as the percentage of nontreated cells in HepG2 cells, was very close to the control. Internalization tests evidenced an intracellular penetration of fluorescent liposomes, but no specific ligand effect was demonstrated ( P > 0.05). Nevertheless, regarding the adenosine triphosphate (ATP) assay, a slight increase was obtained with liposome loaded with ATP incorporating Lac-10-chol after 24 hours ( P < 0.05). [ABSTRACT FROM AUTHOR]

Human natural killer (NK) cells are lymphocytes that destroy tumor cells, virally-infected cells, and antibody-coated cells. Tetrabromobisphenol A (TBBPA) is used both as a reactive and as an additive flame retardant in a variety of materials and appears to contaminate the environment. TBBPA has been found in human blood samples and if it interferes with NK cell function, this could increase the risk of tumor development and/or viral infection. The present study examines the effects of exposure to various concentrations of TBBPA for 24 hr, 48 hr, and 6 days on the lytic function, tumor-target-binding function, and ATP levels of NK cells. These same parameters were also monitored in NK cells that were exposed to TBBPA for 1 h followed by 24 hr, 48 hr, and 6 days in TBBPA-free media. A 24-h exposure of NK cells to 5 μM TBBPA caused a >95% decrease in NK lytic function, a 70% decrease in binding function, and a 34% decrease in ATP levels in NK cells. Exposure to 2.5 μM TBBPA for 24 h decreased lytic function by 76%, binding function by 20%, and had no effect on ATP levels. Exposure of NK cells to 5 μM TBBPA for 1 h followed by 24 h in TBBPA-free media caused a progressive and persistent loss of lytic function (41%) while not affecting either binding ability or ATP levels. The results indicate that TBBPA exposures decrease the lytic function of human NK cells and that an initial brief (1 hr) exposure can cause a progressive loss of function. In addition, the data also indicate that TBBPA-induced loss of NK lytic function can occur at a concentration of TBBPA that does not affect target-binding ability and ATP levels of NK cells. [ABSTRACT FROM AUTHOR]

Autophagy, a lysosomal process involved in the maintenance of cellular homeostasis, is responsible for the turnover of long-lived proteins and organelles that are either damaged or functionally redundant. The process is tightly controlled by the insulin-amino acid-mammalian target of the rapamycin-dependent signal-transduction pathway. Research in the last decade has indicated not only that autophagy provides cells with oxidizable substrate when nutrients become scarce but also that it can provide protection against aging and a number of pathologies such as cancer, neurodegeneration, cardiac disease, diabetes, and infections. [ABSTRACT FROM AUTHOR]

Neuronal accumulation of 1-methyl-4-phenylpyridinium ion (MPP+), the metabolite of neural toxin, 1-methyl-4-phenyl-1,2,3,6-tetrahyropyridine (MPTP), induces a rapid depletion of cellular ATP level and loss of neuronal cell viability which simulates human Parkinson's disease (PD). Since ATP plays an important role in the physiology and function of platelets, which share many biochemical and physiological features with neuronal cells, we examined the effect of MPP+ on platelet aggregation and viability using freshly isolated rat platelets. While the treatment of MPP+ to platelets did not induce cytotoxicity, it significantly attenuated agonist-induced platelet aggregation in a concentration dependent manner. The inhibition of aggregation by MPP+ was mediated by the depletion of the cytoplasmic ATP pool and resultant decreased ATP secretion. Different from the previous reports in neuronal cells, MPP+ did not affect intracellular levels of glutathione and cytoplasmic Ca2+ in platelets. The combined treatment with MPP+ and 2-deoxyglucose, a glycolysis inhibitor, showed the additive effect in the decrease of ATP secretion and intracellular content. Consistent with these findings, inhibitory effects of MPP+ on platelet aggregation was significantly enhanced by the treatment with 2-deoxyglucose. In conclusion, these results suggested that MPP+ can induce ATP depletion in platelets and attenuate platelet aggregation providing a new theory on the reduced platelet activities in PD patients. [ABSTRACT FROM AUTHOR]

Extracellular ATP has been implicated in a number of cellular events, including mammalian sperm function. The complement of ATP-dependent sperm proteins includes six subunits of the 26S proteasome, a multi-subunit protease specific to ubiquitinated substrate-proteins. Proteolysis of ubiquitinated proteins by the 26S proteasome is necessary for the success of mammalian fertilization, including but not limited to acrosomal exocytosis (AE) and sperm-zona pellucida (ZP) penetration. The 26S proteasome is uniquely present on the sperm acrosomal surface during mammalian, ascidian, and invertebrate fertilization. The proteasome is a multi-subunit protease complex of ∼2 MDa composed of the 19S regulatory complex and a 20S proteolytic core. Integrity of the 19S complex is maintained by six 19S ATPase subunits (PSMC1 through PSMC6). Consequently, we hypothesized that fertilization will be blocked by the depletion of sperm-surface associated ATP (ssATP). Depletion of ssATP by the Solanum tuberosum apyrase, a 49 kDa, non-cell permeant enzyme, significantly reduced the ATP content measured by an adapted luminescence-ATP assay from which all permeabilizing agents were excluded. Addition of active apyrase to porcine in vitro fertilization (IVF) medium caused a concentration dependent reduction in the overall fertilization rate. No such outcomes were observed in control groups using heat-inactivated apyrase. Apyrase treatment altered the band pattern of 19S ATPase subunits PSMC1 (Rpt2) and PSMC4 (Rpt3) in Western blotting, suggesting that it had an effect on the integrity of the sperm proteasomal 19S complex. Apyrase only altered the proteasomal core activities slightly, since these activities are not directly dependent on external ATP. In contrast, sperm treatment with MG132, a specific inhibitor of the proteasomal core chymotrypsin-like activity, inhibited the target proteolytic activity, but also induced a compensatory elevation in proteasomal peptidyl-glutamyl peptide hydrolase activity. Altogether, the present data provide an important missing piece of evidence in support of the ssATP-dependent, proteasomal-proteolytic model of sperm-ZP interactions. [ABSTRACT FROM AUTHOR]

Platelets play a critical role in homeostasis and blood clotting at sites of vascular injury, and also in various ways in innate immunity and inflammation. Platelets are one of the first cells to accumulate at an injured site, and local release of their secretome at some point initiate an inflammatory cascade that attracts leukocytes, activates target cells, stimulates vessel growth and repair. The level of exogenous ATP in the body may be increased in various inflammatory and shock conditions, primarily as a consequence of nucleotide release from platelets, endothelium and blood vessel cells. An increase of ATP release has been described during inflammation and this compound presents proinflammatory properties. ADP is a nucleotide known to induce changes in platelets shape and aggregation, to promote the exposure of fibrinogen-binding sites and to inhibit the stimulation of adenylate cyclase. Adenosine, the final product of the nucleotide hydrolysis, is a vasodilator and an inhibitor of platelet aggregation. There is a group of ecto-enzymes responsible for extracellular nucleotide hydrolysis named ectonucleotidases, which includes the NTPDase (nucleoside triphosphate diphosphohydrolase) family, the NPP (nucleoside pyrophosphatase/phosphodiesterase) family and an ecto-5'-nucleotidase. Therefore, we have aimed to investigate the effect of lipopolysaccharide endotoxin from Escherichia coli on ectonucleotidases in platelets from adult rats in order to better understand the role of extracellular adenine nucleotides and nucleosides in the maintenance of blood homeostasis in inflammatory processes. LPS administered in vitro was not able to alter the ATP, ADP, AMP and ρ-Nph-5'-TMP hydrolysis of platelets from untreated rats in all concentrations tested (25-100 μg/ml). There was a significant decrease in ATP, ADP, AMP and ρ-Nph-5'-TMP hydrolysis in rat platelets after 48 hours of LPS exposure (2 mg/Kg, i.p.). ATP and ADP hydrolysis has been reduced about 28% whereas it has been observed a significant 30% and 26% decrease on AMP and ρ-Nph-5'-TMP hydrolysis. Platelet aggregation and platelet number have shown a significant decrease in LPS-treated rats (40% and 55%, respectively) when compared to control group. These results suggest that changes observed in platelet count and, consequently, in nucleotidase activities from circulatory system could alter extracellular nucleotide and nucleoside levels, which might modulate the inflammatory process. [ABSTRACT FROM AUTHOR]

The herbicide bentazon (CASRN 25057-89-0) is extensively used in agriculture in Brittany (France) to replace atrazine. Bentazon is not readily adsorbed by soil and therefore it enters adjacent freshwater ecosystems, making its way to estuarine and marine waters areas. Information regarding its effects on marine ecosystems is scarce. Phytotoxicity assessments were conducted in the laboratory on the common diatom Chaetoceros gracilis using both the active ingredient and its formulated product (Basamais). The 3 day EC50 using cell counts were, respectively, 150 mg/L and 60 μg/L for bentazon active ingredient and for bentazon-formulated while cellular volume was increased. Although bentazon is known as a photosystem II inhibitor, it produced an increase of pigment (chlorophylls a, c, and carotenoids) content, ATP synthesis, rates of photosynthesis and respiration, and TBARS formation. Therefore, pigments cannot be used as biomarkers of toxicity. Algal cell recovery from bentazon effects occurred after 6-9 days, suggesting a tolerance mechanism. [ABSTRACT FROM AUTHOR]

In this work, Diisopropyl [phenyl(pyridine-4-carboxamido)methyl]phosphonate was synthesized for investigating the interaction with ATP. The noncovalent complex was observed by the Electrospray ionization Mass Spectrometry (ESI-MS). Through Molecular modeling, it was found that the non-covalent complex was stabilized by two intermolecular hydrogen bonds, which has the binding energy of -8.999 kcals/mol. In order to investigate the function of phosphoryl group in the interaction, benzoyl phenylglycine isopropyl ester was also tested, but no interaction with ATP was observed by ESI-MS. These results implied that phosphoryl group was a very important functional group to provide effective interaction site between α-Aminophosphonic acids and ATP. [ABSTRACT FROM AUTHOR]