WorldWideScience

Sample records for high atp concentrations

  1. Exposure to high glutamate concentration activates aerobic glycolysis but inhibits ATP-linked respiration in cultured cortical astrocytes.

    Science.gov (United States)

    Shen, Yao; Tian, Yueyang; Shi, Xiaojie; Yang, Jianbo; Ouyang, Li; Gao, Jieqiong; Lu, Jianxin

    2014-08-01

    Astrocytes play a key role in removing the synaptically released glutamate from the extracellular space and maintaining the glutamate below neurotoxic level in the brain. However, high concentration of glutamate leads to toxicity in astrocytes, and the underlying mechanisms are unclear. The purpose of this study was to investigate whether energy metabolism disorder, especially impairment of mitochondrial respiration, is involved in the glutamate-induced gliotoxicity. Exposure to 10-mM glutamate for 48 h stimulated glycolysis and respiration in astrocytes. However, the increased oxygen consumption was used for proton leak and non-mitochondrial respiration, but not for oxidative phosphorylation and ATP generation. When the exposure time extended to 72 h, glycolysis was still activated for ATP generation, but the mitochondrial ATP-linked respiration of astrocytes was reduced. The glutamate-induced astrocyte damage can be mimicked by the non-metabolized substrate d-aspartate but reversed by the non-selective glutamate transporter inhibitor TBOA. In addition, the glutamate toxicity can be partially reversed by vitamin E. These findings demonstrate that changes of bioenergetic profile occur in cultured cortical astrocytes exposed to high concentration of glutamate and highlight the role of mitochondria respiration in glutamate-induced gliotoxicity in cortical astrocytes. Copyright © 2014 John Wiley & Sons, Ltd.

  2. Role of glycogenolysis in stimulation of ATP release from cultured mouse astrocytes by transmitters and high K+ concentrations

    Directory of Open Access Journals (Sweden)

    Junnan Xu

    2014-01-01

    Full Text Available This study investigates the role of glycogenolysis in stimulated release of ATP as a transmitter from astrocytes. Within the last 20 years our understanding of brain glycogenolysis has changed from it being a relatively uninteresting process to being a driving force for essential brain functions like production of transmitter glutamate and homoeostasis of potassium ions (K+ after their release from excited neurons. Simultaneously, the importance of astrocytic handling of adenosine, its phosphorylation to ATP and release of some astrocytic ATP, located in vesicles, as an important transmitter has also become to be realized. Among the procedures stimulating Ca2+-dependent release of vesicular ATP are exposure to such transmitters as glutamate and adenosine, which raise intra-astrocytic Ca2+ concentration, or increase of extracellular K+ to a depolarizing level that opens astrocytic L-channels for Ca2+ and thereby also increase intra-astrocytic Ca2+ concentration, a prerequisite for glycogenolysis. The present study has confirmed and quantitated stimulated ATP release from well differentiated astrocyte cultures by glutamate, adenosine or elevated extracellular K+ concentrations, measured by a luciferin/luciferase reaction. It has also shown that this release is virtually abolished by an inhibitor of glycogenolysis as well as by inhibitors of transmitter-mediated signaling or of L-channel opening by elevated K+ concentrations.

  3. Role of Glycogenolysis in Stimulation of ATP Release from Cultured Mouse Astrocytes by Transmitters and High K+ Concentrations

    Directory of Open Access Journals (Sweden)

    Junnan Xu

    2013-12-01

    Full Text Available This study investigates the role of glycogenolysis in stimulated release of ATP as a transmitter from astrocytes. Within the last 20 years our understanding of brain glycogenolysis has changed from it being a relatively uninteresting process to being a driving force for essential brain functions like production of transmitter glutamate and homoeostasis of potassium ions (K+ after their release from excited neurons. Simultaneously, the importance of astrocytic handling of adenosine, its phosphorylation to ATP and release of some astrocytic ATP, located in vesicles, as an important transmitter has also become to be realized. Among the procedures stimulating Ca2+ -dependent release of vesicular ATP are exposure to such transmitters as glutamate and adenosine, which raise intra-astrocytic Ca2+ concentration, or increase of extracellular K+ to a depolarizing level that opens astrocytic L-channels for Ca2+ and thereby also increase intra-astrocytic Ca2+ concentration, a prerequisite for glycogenolysis. The present study has confirmed and quantitated stimulated ATP release from well differentiated astrocyte cultures by glutamate, adenosine or elevated extracellular K+ concentrations, measured by a luciferin/luciferase reaction. It has also shown that this release is virtually abolished by an inhibitor of glycogenolysis as well as by inhibitors of transmitter-mediated signaling or of L-channel opening by elevated K+ concentrations.

  4. Role of glycogenolysis in stimulation of ATP release from cultured mouse astrocytes by transmitters and high K+ concentrations.

    Science.gov (United States)

    Xu, Junnan; Song, Dan; Bai, Qiufang; Zhou, Lijun; Cai, Liping; Hertz, Leif; Peng, Liang

    2014-01-13

    This study investigates the role of glycogenolysis in stimulated release of ATP as a transmitter from astrocytes. Within the last 20 years our understanding of brain glycogenolysis has changed from it being a relatively uninteresting process to being a driving force for essential brain functions like production of transmitter glutamate and homoeostasis of potassium ions (K+) after their release from excited neurons. Simultaneously, the importance of astrocytic handling of adenosine, its phosphorylation to ATP and release of some astrocytic ATP, located in vesicles, as an important transmitter has also become to be realized. Among the procedures stimulating Ca2+-dependent release of vesicular ATP are exposure to such transmitters as glutamate and adenosine, which raise intra-astrocytic Ca2+ concentration, or increase of extracellular K+ to a depolarizing level that opens astrocytic L-channels for Ca2+ and thereby also increase intra-astrocytic Ca2+ concentration, a prerequisite for glycogenolysis. The present study has confirmed and quantitated stimulated ATP release from well differentiated astrocyte cultures by glutamate, adenosine or elevated extracellular K+ concentrations, measured by a luciferin/luciferase reaction. It has also shown that this release is virtually abolished by an inhibitor of glycogenolysis as well as by inhibitors of transmitter-mediated signaling or of L-channel opening by elevated K+ concentrations.

  5. Dynamic imaging of free cytosolic ATP concentration during fuel sensing by rat hypothalamic neurones: evidence for ATP-independent control of ATP-sensitive K(+) channels.

    Science.gov (United States)

    Ainscow, Edward K; Mirshamsi, Shirin; Tang, Teresa; Ashford, Michael L J; Rutter, Guy A

    2002-10-15

    Glucose-responsive (GR) neurons from hypothalamic nuclei are implicated in the regulation of feeding and satiety. To determine the role of intracellular ATP in the closure of ATP-sensitive K(+) (K(ATP)) channels in these cells and associated glia, the cytosolic ATP concentration ([ATP](c)) was monitored in vivo using adenoviral-driven expression of recombinant targeted luciferases and bioluminescence imaging. Arguing against a role for ATP in the closure of K(ATP) channels in GR neurons, glucose (3 or 15 mM) caused no detectable increase in [ATP](c), monitored with cytosolic luciferase, and only a small decrease in the concentration of ATP immediately beneath the plasma membrane, monitored with a SNAP25-luciferase fusion protein. In contrast to hypothalamic neurons, hypothalamic glia responded to glucose (3 and 15 mM) with a significant increase in [ATP](c). Both neurons and glia from the cerebellum, a glucose-unresponsive region of the brain, responded robustly to 3 or 15 mM glucose with increases in [ATP](c). Further implicating an ATP-independent mechanism of K(ATP) channel closure in hypothalamic neurons, removal of extracellular glucose (10 mM) suppressed the electrical activity of GR neurons in the presence of a fixed, high concentration (3 mM) of intracellular ATP. Neurons from both brain regions responded to 5 mM lactate (but not pyruvate) with an oligomycin-sensitive increase in [ATP](c). High levels of the plasma membrane lactate-monocarboxylate transporter, MCT1, were found in both cell types, and exogenous lactate efficiently closed K(ATP) channels in GR neurons. These data suggest that (1) ATP-independent intracellular signalling mechanisms lead to the stimulation of hypothalamic neurons by glucose, and (2) these effects may be potentiated in vivo by the release of lactate from neighbouring glial cells.

  6. Dynamic imaging of free cytosolic ATP concentration during fuel sensing by rat hypothalamic neurones: evidence for ATP-independent control of ATP-sensitive K+ channels

    Science.gov (United States)

    Ainscow, Edward K; Mirshamsi, Shirin; Tang, Teresa; Ashford, Michael L J; Rutter, Guy A

    2002-01-01

    Glucose-responsive (GR) neurons from hypothalamic nuclei are implicated in the regulation of feeding and satiety. To determine the role of intracellular ATP in the closure of ATP-sensitive K+ (KATP) channels in these cells and associated glia, the cytosolic ATP concentration ([ATP]c) was monitored in vivo using adenoviral-driven expression of recombinant targeted luciferases and bioluminescence imaging. Arguing against a role for ATP in the closure of KATP channels in GR neurons, glucose (3 or 15 mm) caused no detectable increase in [ATP]c, monitored with cytosolic luciferase, and only a small decrease in the concentration of ATP immediately beneath the plasma membrane, monitored with a SNAP25–luciferase fusion protein. In contrast to hypothalamic neurons, hypothalamic glia responded to glucose (3 and 15 mm) with a significant increase in [ATP]c. Both neurons and glia from the cerebellum, a glucose-unresponsive region of the brain, responded robustly to 3 or 15 mm glucose with increases in [ATP]c. Further implicating an ATP-independent mechanism of KATP channel closure in hypothalamic neurons, removal of extracellular glucose (10 mm) suppressed the electrical activity of GR neurons in the presence of a fixed, high concentration (3 mm) of intracellular ATP. Neurons from both brain regions responded to 5 mm lactate (but not pyruvate) with an oligomycin-sensitive increase in [ATP]c. High levels of the plasma membrane lactate-monocarboxylate transporter, MCT1, were found in both cell types, and exogenous lactate efficiently closed KATP channels in GR neurons. These data suggest that (1) ATP-independent intracellular signalling mechanisms lead to the stimulation of hypothalamic neurons by glucose, and (2) these effects may be potentiated in vivo by the release of lactate from neighbouring glial cells. PMID:12381816

  7. Role of glycogenolysis in stimulation of ATP release from cultured mouse astrocytes by transmitters and high K+ concentrations

    OpenAIRE

    Junnan Xu; Dan Song; Qiufang Bai; Lijun Zhou; Liping Cai; Leif Hertz; Liang Peng

    2014-01-01

    This study investigates the role of glycogenolysis in stimulated release of ATP as a transmitter from astrocytes. Within the last 20 years our understanding of brain glycogenolysis has changed from it being a relatively uninteresting process to being a driving force for essential brain functions like production of transmitter glutamate and homoeostasis of potassium ions (K+) after their release from excited neurons. Simultaneously, the importance of astrocytic handling of adenosine, its phosp...

  8. Use of an ATP-based chemosensitivity assay to design new combinations of high-concentration doxorubicin with other drugs for recurrent ovarian cancer.

    Science.gov (United States)

    Di Nicolantonio, Federica; Neale, Michael H; Knight, Louise A; Lamont, Alan; Skailes, Geraldine E; Osborne, Richard J; Allerton, Rosanne; Kurbacher, Christian M; Cree, Ian A

    2002-07-01

    Liposomal doxorubicin (Caelyx/Doxil) has been shown to be active in around 20% of recurrent ovarian cancers. As yet, there is little clinical data on combinations of existing agents with liposomal doxorubicin, despite considerable clinical experience with soluble doxorubicin in combination. In this study, we have used an ATP-based tumor chemosensitivity assay to determine the relative efficacy of high concentrations of doxorubicin tested in combination with cisplatin, treosulfan, 5-fluorouracil (5-FU) or vinorelbine against cells obtained from recurrent ovarian tumor tissue. The results show little enhancement of the efficacy of high concentrations of doxorubicin by 5-FU, cisplatin, or treosulfan. However, vinorelbine+liposomal doxorubicin showed additive inhibition, and this combination is worthy of further testing in clinical trials.

  9. Muscle interstitial ATP and norepinephrine concentrations in the human leg during exercise and ATP infusion

    DEFF Research Database (Denmark)

    Mortensen, Stefan P.; Gonzalez-Alonso, Jose; Nielsen, Jens Jung

    2009-01-01

    ATP has been proposed to play multiple roles in local skeletal muscle blood flow regulation by inducing vasodilation and modulating sympathetic vasoconstrictor activity, but the mechanism remain unclear. Here we evaluated the effects of arterial ATP infusion and exercise on limb muscle interstitial...... ATP and NE concentrations to gain insight into the interstitial and intravascular mechanisms by which ATP causes muscle vasodilation and sympatholysis. Leg hemodynamics and muscle interstitial nucleotide and norepinephrine (NE) concentrations were measured during: 1) femoral arterial ATP infusion (0.......42+/-0.04 and 2.26+/-0.52 mumol/min; mean+/-SEM) and 2) one-leg knee-extensor exercise (18+/-0 and 37+/-2W) in 10 healthy, male subjects. Arterial ATP infusion and exercise increased leg blood flow (LBF) in the experimental leg from ~0.3 L/min at baseline to 4.2+/-0.3 and 4.6+/-0.5 L/min, respectively, whereas...

  10. Sensitivity of small myosin II ensembles from different isoforms to mechanical load and ATP concentration

    Science.gov (United States)

    Erdmann, Thorsten; Bartelheimer, Kathrin; Schwarz, Ulrich S.

    2016-11-01

    Based on a detailed crossbridge model for individual myosin II motors, we systematically study the influence of mechanical load and adenosine triphosphate (ATP) concentration on small myosin II ensembles made from different isoforms. For skeletal and smooth muscle myosin II, which are often used in actomyosin gels that reconstitute cell contractility, fast forward movement is restricted to a small region of phase space with low mechanical load and high ATP concentration, which is also characterized by frequent ensemble detachment. At high load, these ensembles are stalled or move backwards, but forward motion can be restored by decreasing ATP concentration. In contrast, small ensembles of nonmuscle myosin II isoforms, which are found in the cytoskeleton of nonmuscle cells, are hardly affected by ATP concentration due to the slow kinetics of the bound states. For all isoforms, the thermodynamic efficiency of ensemble movement increases with decreasing ATP concentration, but this effect is weaker for the nonmuscle myosin II isoforms.

  11. Correlation of microbial mass with ATP and DNA concentrations in acidogenesis of whey permeate.

    Science.gov (United States)

    Kim, Jaai; Lee, Changsoo; Shin, Seung Gu; Hwang, Seokhwan

    2008-04-01

    In this paper, we examine variations in the contents of ATP and DNA per unit microbial mass in an acidogenesis of whey permeate. We also introduce a novel approach to estimate microbial mass by measuring ATP and DNA when the ratios of ATP and DNA to microbial mass vary. Acidogenic experiments were performed at 35 degrees C and pH 6.0 in batch mode. The amounts of ATP and DNA per unit microbial mass were not consistent during the incubation except during the post-decay phase. Especially within the exponential phase, each showed a 10-fold difference between maximal and minimal values. In this case, the conventional method which converts ATP or DNA concentration into microbial mass using a fixed conversion factor can give inaccurate results. While the constant ratios of 0.74 mg ATP/g VSS and 1.96 mg DNA/g VSS were determined for the post-decay phase, the ATP and DNA concentrations showed strong linear relationships with the microbial mass (r2=0.99) within the ranges of 0.039-1.078 mg ATP/l and 0.075-2.080 mg DNA/l, respectively. The linear regression equations are as follows: (1) microbial mass concentration (mg/l)=478.5xATP concentration (mg/l)+293.5, (2) microbial mass concentration (mg/l)=257.2xDNA concentration (mg/l)+250.4. Therefore, changes in the mass of the acidogenic population should be monitored by the combined use of the regression equations obtained in the exponential phase and the constant ratios determined in the post-decay phase. This procedure should be widely applicable to the acidogenesis of dairy processing wastewaters, especially of a highly suspended organic wastewater such as whey.

  12. An exonuclease I-based label-free fluorometric aptasensor for adenosine triphosphate (ATP) detection with a wide concentration range.

    Science.gov (United States)

    Wei, Yanli; Chen, Yanxia; Li, Huanhuan; Shuang, Shaomin; Dong, Chuan; Wang, Gufeng

    2015-01-15

    A novel aptamer-based label-free assay for sensitive and selective detection of ATP was developed. This assay employs a new aptamer/fluorescent probe system that shows resistance to exonuclease I (Exo I) digestion upon binding to ATP molecules. In the absence of ATP, the complex between the ATP-binding aptamer (ATP-aptamer) and a DNA binding dye, berberine, is digested upon the addition of exonuclease I, leading to the release of berberine into solution and consequently, quenched berberine fluorescence. In the presence of ATP, the ATP-binding aptamer folds into a G-quadruplex structure that is resistant to Exo I digestion. Accordingly, berberine is protected in the G-quadruplex structure and high fluorescence intensity is observed. As such, based on the fluorescence signal change, a label-free fluorescence assay for ATP was developed. Factors affecting the analysis of ATP including the concentration of ATP-binding aptamer, reaction time, temperature and the concentration of Exo I were comprehensively investigated. Under optimal conditions, the fluorescence intensity of the sensing system displayed a response for ATP in a wide range up to 17.5 mM with a detection limit of 140 nM.

  13. Extracellular ATP inhibits root gravitropism at concentrations that inhibit polar auxin transport

    Science.gov (United States)

    Tang, Wenqiang; Brady, Shari R.; Sun, Yu; Muday, Gloria K.; Roux, Stanley J.

    2003-01-01

    Raising the level of extracellular ATP to mM concentrations similar to those found inside cells can block gravitropism of Arabidopsis roots. When plants are grown in Murashige and Skoog medium supplied with 1 mM ATP, their roots grow horizontally instead of growing straight down. Medium with 2 mM ATP induces root curling, and 3 mM ATP stimulates lateral root growth. When plants are transferred to medium containing exogenous ATP, the gravity response is reduced or in some cases completely blocked by ATP. Equivalent concentrations of ADP or inorganic phosphate have slight but usually statistically insignificant effects, suggesting the specificity of ATP in these responses. The ATP effects may be attributable to the disturbance of auxin distribution in roots by exogenously applied ATP, because extracellular ATP can alter the pattern of auxin-induced gene expression in DR5-beta-glucuronidase transgenic plants and increase the response sensitivity of plant roots to exogenously added auxin. The presence of extracellular ATP also decreases basipetal auxin transport in a dose-dependent fashion in both maize (Zea mays) and Arabidopsis roots and increases the retention of [(3)H]indole-3-acetic acid in root tips of maize. Taken together, these results suggest that the inhibitory effects of extracellular ATP on auxin distribution may happen at the level of auxin export. The potential role of the trans-plasma membrane ATP gradient in auxin export and plant root gravitropism is discussed.

  14. Modeling Interactions among Individual P2 Receptors to Explain Complex Response Patterns over a Wide Range of ATP Concentrations.

    Science.gov (United States)

    Xing, Shu; Grol, Matthew W; Grutter, Peter H; Dixon, S Jeffrey; Komarova, Svetlana V

    2016-01-01

    Extracellular ATP acts on the P2X family of ligand-gated ion channels and several members of the P2Y family of G protein-coupled receptors to mediate intercellular communication among many cell types including bone-forming osteoblasts. It is known that multiple P2 receptors are expressed on osteoblasts (P2X2,5,6,7 and P2Y1,2,4,6). In the current study, we investigated complex interactions within the P2 receptor network using mathematical modeling. To characterize individual P2 receptors, we extracted data from published studies of overexpressed human and rodent (rat and mouse) receptors and fit their dependencies on ATP concentration using the Hill equation. Next, we examined responses induced by an ensemble of endogenously expressed P2 receptors. Murine osteoblastic cells (MC3T3-E1 cells) were loaded with fluo-4 and stimulated with varying concentrations of extracellular ATP. Elevations in the concentration of cytosolic free calcium ([Ca(2+)]i) were monitored by confocal microscopy. Dependence of the calcium response on ATP concentration exhibited a complex pattern that was not explained by the simple addition of individual receptor responses. Fitting the experimental data with a combination of Hill equations from individual receptors revealed that P2Y1 and P2X7 mediated the rise in [Ca(2+)]i at very low and high ATP concentrations, respectively. Interestingly, to describe responses at intermediate ATP concentrations, we had to assume that a receptor with a K 1∕2 in that range (e.g. P2Y4 or P2X5) exerts an inhibitory effect. This study provides new insights into the interactions among individual P2 receptors in producing an ensemble response to extracellular ATP.

  15. Modeling interactions among individual P2 receptors to explain complex response patterns over a wide range of ATP concentrations

    Directory of Open Access Journals (Sweden)

    Shu Xing

    2016-07-01

    Full Text Available EExtracellular ATP acts on the P2X family of ligand-gated ion channels and several members of the P2Y family of G protein-coupled receptors to mediate intercellular communication among many cell types including bone-forming osteoblasts. It is known that multiple P2 receptors are expressed on osteoblasts (P2X2,5,6,7 and P2Y1,2,4,6. In the current study, we investigated complex interactions within the P2 receptor network using mathematical modeling. To characterize individual P2 receptors, we extracted data from published studies of overexpressed human and rodent (rat and mouse receptors and fit their dependencies on ATP concentration using the Hill equation. Next, we examined responses induced by an ensemble of endogenously expressed P2 receptors. Murine osteoblastic cells (MC3T3-E1 cells were loaded with fluo-4 and stimulated with varying concentrations of extracellular ATP. Elevations in the concentration of cytosolic free calcium ([Ca2+]i were monitored by confocal microscopy. Dependence of the calcium response on ATP concentration exhibited a complex pattern that was not explained by the simple addition of individual receptor responses. Fitting the experimental data with a combination of Hill equations from individual receptors revealed that P2Y1 and P2X7 mediated the rise in [Ca2+]i at very low and high ATP concentrations, respectively. Interestingly, to describe responses at intermediate ATP concentrations, we had to assume that a receptor with a K1/2 in that range (e.g. P2Y4 or P2X5 exerts an inhibitory effect. This study provides new insights into the interactions among individual P2 receptors in producing an ensemble response to extracellular ATP.

  16. Plasma ATP concentration and venous oxygen content in the forearm during dynamic handgrip exercise

    Directory of Open Access Journals (Sweden)

    Askew Christopher D

    2009-12-01

    Full Text Available Abstract Background It has been proposed that adenosine triphosphate (ATP released from red blood cells (RBCs may contribute to the tight coupling between blood flow and oxygen demand in contracting skeletal muscle. To determine whether ATP may contribute to the vasodilatory response to exercise in the forearm, we measured arterialised and venous plasma ATP concentration and venous oxygen content in 10 healthy young males at rest, and at 30 and 180 seconds during dynamic handgrip exercise at 45% of maximum voluntary contraction (MVC. Results Venous plasma ATP concentration was elevated above rest after 30 seconds of exercise (P Conclusions Collectively these results indicate that ATP in the plasma originated from the muscle microcirculation, and are consistent with the notion that deoxygenation of the blood perfusing the muscle acts as a stimulus for ATP release. That ATP concentration was elevated just 30 seconds after the onset of exercise also suggests that ATP may be a contributing factor to the blood flow response in the transition from rest to steady state exercise.

  17. Acetoacetate reduces growth and ATP concentration in cancer cell lines which over-express uncoupling protein 2

    Directory of Open Access Journals (Sweden)

    Quadros Edward V

    2009-05-01

    Full Text Available Abstract Background Recent evidence suggests that several human cancers are capable of uncoupling of mitochondrial ATP generation in the presence of intact tricarboxylic acid (TCA enzymes. The goal of the current study was to test the hypothesis that ketone bodies can inhibit cell growth in aggressive cancers and that expression of uncoupling protein 2 is a contributing factor. The proposed mechanism involves inhibition of glycolytic ATP production via a Randle-like cycle while increased uncoupling renders cancers unable to produce compensatory ATP from respiration. Methods Seven aggressive human cancer cell lines, and three control fibroblast lines were grown in vitro in either 10 mM glucose medium (GM, or in glucose plus 10 mM acetoacetate [G+AcA]. The cells were assayed for cell growth, ATP production and expression of UCP2. Results There was a high correlation of cell growth with ATP concentration (r = 0.948 in a continuum across all cell lines. Controls demonstrated normal cell growth and ATP with the lowest density of mitochondrial UCP2 staining while all cancer lines demonstrated proportionally inhibited growth and ATP, and over-expression of UCP2 (p Conclusion Seven human cancer cell lines grown in glucose plus acetoacetate medium showed tightly coupled reduction of growth and ATP concentration. The findings were not observed in control fibroblasts. The observed over-expression of UCP2 in cancer lines, but not in controls, provides a plausible molecular mechanism by which acetoacetate spares normal cells but suppresses growth in cancer lines. The results bear on the hypothesized potential for ketogenic diets as therapeutic strategies.

  18. Highly Selective Cerebral ATP Assay Based on Micrometer Scale Ion Current Rectification at Polyimidazolium-Modified Micropipettes.

    Science.gov (United States)

    Zhang, Kailin; He, Xiulan; Liu, Yang; Yu, Ping; Fei, Junjie; Mao, Lanqun

    2017-06-20

    Development of new principles and methods for cerebral ATP assay is highly imperative not only for determining ATP dynamics in brain but also for understanding physiological and pathological processes related to ATP. Herein, we for the first time demonstrate that micrometer scale ion current rectification (MICR) at a polyimidazolium brush-modified micropipette can be used as the signal transduction output for the cerebral ATP assay with a high selectivity. The rationale for ATP assay is essentially based on the competitive binding ability between positively charged polyimidazolium and ATP toward negatively charged ATP aptamer. The method is well responsive to ATP with a good linearity within a concentration range from 5 nM to 100 nM, and high selectivity toward ATP. These properties essentially enable the method to determine the cerebral ATP by combining in vivo microdialysis. The basal dialysate level of ATP in rat brain cortex is determined to be 11.32 ± 2.36 nM (n = 3). This study demonstrates that the MICR-based sensors could be potentially used for monitoring neurochemicals in cerebral systems.

  19. Measuring ATP Concentration in a Small Number of Murine Hematopoietic Stem Cells.

    Science.gov (United States)

    Szade, Krzysztof; Zukowska, Monika; Jozkowicz, Alicja; Dulak, Jozef

    2016-01-01

    The metabolism of quiescent adult stem cells differs from the metabolism of differentiated cells. The metabolic processes are tightly regulated and their alterations disturb function of stem cells. One of the indicators of metabolic status of cells is the ATP level. While the method of measuring the ATP levels has been known for many years, estimating ATP levels in small population of defined stem cells isolated directly from the tissue has remained challenging. Here, we show our method of measuring the ATP levels in hematopoietic stem cells sorted from murine bone marrow. We used magnetic sorting as well as cell sorter and adopted the commonly used bioluminescence-based detection kits in described protocol. Our strategy allows to measure ATP levels in 1000 highly purified HSC.

  20. The distribution of ATP within tomato (Lycopersicon esculentum Mill.) embryos correlates with germination whee as total ATP concentration does not

    NARCIS (Netherlands)

    Spoelstra, P.; Joosen, R.V.L.; Hilhorst, H.W.M.

    2002-01-01

    The distribution of ATP in tomato seeds was visualized by monitoring the luminescence of frozen sections on top of a gel containing all the components of the luciferase reaction, but excluding ATP. ATP was imaged in germinating tomato seeds at intervals of 3, 6, 17, 24 and 48 h and in seeds with pri

  1. The distribution of ATP within tomato (Lycopersicon esculentum Mill.) embryos correlates with germination whee as total ATP concentration does not

    NARCIS (Netherlands)

    Spoelstra, P.; Joosen, R.V.L.; Hilhorst, H.W.M.

    2002-01-01

    The distribution of ATP in tomato seeds was visualized by monitoring the luminescence of frozen sections on top of a gel containing all the components of the luciferase reaction, but excluding ATP. ATP was imaged in germinating tomato seeds at intervals of 3, 6, 17, 24 and 48 h and in seeds with

  2. BTeam, a Novel BRET-based Biosensor for the Accurate Quantification of ATP Concentration within Living Cells

    Science.gov (United States)

    Yoshida, Tomoki; Kakizuka, Akira; Imamura, Hiromi

    2016-01-01

    ATP levels may represent fundamental health conditions of cells. However, precise measurement of intracellular ATP levels in living cells is hindered by the lack of suitable methodologies. Here, we developed a novel ATP biosensor termed “BTeam”. BTeam comprises a yellow fluorescent protein (YFP), the ATP binding domain of the ε subunit of the bacterial ATP synthase, and an ATP-nonconsuming luciferase (NLuc). To attain emission, BTeam simply required NLuc substrate. BTeam showed elevated bioluminescence resonance energy transfer efficiency upon ATP binding, resulted in the emission spectra changes correlating with ATP concentrations. By using values of YFP/NLuc emission ratio to represent ATP levels, BTeam achieved steady signal outputs even though emission intensities were altered. With this biosensor, we succeeded in the accurate quantification of intracellular ATP concentrations of a population of living cells, as demonstrated by detecting the slight distribution in the cytosol (3.7–4.1 mM) and mitochondrial matrix (2.4–2.7 mM) within some cultured cell lines. Furthermore, BTeam allowed continuous tracing of cytosolic ATP levels of the same cells, as well as bioluminescent imaging of cytosolic ATP dynamics within individual cells. This simple and accurate technique should be an effective method for quantitative measurement of intracellular ATP concentrations. PMID:28000761

  3. Clemastine Potentiates the Human P2X7 Receptor by Sensitizing It to Lower ATP Concentrations*

    OpenAIRE

    Nörenberg, Wolfgang; Hempel, Christoph; Urban, Nicole; Sobottka, Helga; Illes, Peter; Schaefer, Michael

    2011-01-01

    P2X7 receptors have emerged as potential drug targets for the treatment of medical conditions such as e.g. rheumatoid arthritis and neuropathic pain. To assess the impact of pharmaceuticals on P2X7, we screened a compound library comprising approved or clinically tested drugs and identified several compounds that augment the ATP-triggered P2X7 activity in a stably transfected HEK293 cell line. Of these, clemastine markedly sensitized Ca2+ entry through P2X7 to lower ATP concentrations. Extrac...

  4. Sodium (Na+) concentration effects on metabolic pathway and estimation of ATP use in dark fermentation hydrogen production through stoichiometric analysis.

    Science.gov (United States)

    Lee, Myoung-Joo; Kim, Tae-Hyeong; Min, Booki; Hwang, Sun-Jin

    2012-10-15

    Batch experiments were conducted to investigate the effects of Na(+) concentration on hydrogen production with dark fermentation. The Na(+) concentration was varied from 0 to 8 g/L in the mixed culture using an anaerobic sludge treated by acid. The maximum hydrogen production was achieved with 1 g-Na(+)/L, whereas the hydrogen production was decreased over 2 g-Na(+)/L due to the inhibitory of Na(+). The mechanisms of Na(+) inhibition to the hydrogen production are studied using pure culture of Clostridium butyricum by calculating the energy balance. At a high sodium concentration, C. butyricum used a greater proportion of the ATP generated via fermentation for cell maintenance rather than for cell synthesis. Additionally, higher Na(+) concentrations shifted the fermentation process toward the acetate synthesis pathway instead of the butyrate pathway, and the value of Y(X/ATP) decreased. With high Na(+) concentrations, a greater ratio of hydrogen was produced via the oxidation of NADH. Excess hydrogen production decreased as the Na(+) concentration increased.

  5. Highly Divergent Mitochondrial ATP Synthase Complexes in Tetrahymena thermophila

    NARCIS (Netherlands)

    Nina, Praveen Balabaskaran; Dudkina, Natalya V.; Kane, Lesley A.; van Eyk, Jennifer E.; Boekema, Egbert J.; Mather, Michael W.; Vaidya, Akhil B.; Eisen, Jonathan A.

    2010-01-01

    The F-type ATP synthase complex is a rotary nano-motor driven by proton motive force to synthesize ATP. Its F(1) sector catalyzes ATP synthesis, whereas the F(o) sector conducts the protons and provides a stator for the rotary action of the complex. Components of both F(1) and F(o) sectors are highl

  6. Isolation of ATP from a yeast fermentation broth using a cryogel column at high flow velocities.

    Science.gov (United States)

    Yan, Chen; Shen, Shaochuan; Yun, Junxian; Wang, Lianghua; Yao, Kejian; Yao, Shan-Jing

    2008-12-01

    This communication presents an effective method for isolating adenosine triphosphate (ATP) from a yeast fermentation broth using an anion-exchange supermacroporous cryogel column at high flow velocities. The breakthrough and elution behaviors of pure ATP in the cryogel bed were investigated at flow velocities of 2, 5, and 10 cm/min and the ATP binding capacities were determined. Then the ATP-containing yeast fermentation broth was employed as the test feedstock and various chromatographic runs were conducted to isolate ATP by the cryogel at different high flow velocities. The ATP samples obtained were analyzed quantitatively by HPLC. The results showed that even at a flow velocity of 5 or 10 cm/min, a product purity of 97.4 or 98.0% can be achieved, illustrating the potential of the present method for separation of high-purity ATP directly from fermentation feedstock at high flow velocities.

  7. ATP concentration as possible marker of liver damage at leukaemia treatment: confocal microscopy-based experimental study and numerical simulations

    Science.gov (United States)

    Malashchenko, V.; Zyubin, A.; Babak, S.; Lavrova, A.

    2017-04-01

    We consider the method of confocal microscopy as a convenient instrument for determination of chemical compounds in biological tissues and cells. In particular, we study the dynamics of adenosine triphosphate (ATP) concentration that could be used as a bio-marker of energy metabolism pathologies at the treatment of acute lymphoblastic leukaemia (ALL). On the basis of data obtained by the confocal microscopy, the values of ATP concentration have been calculated for each case. Possible correlations with other characteristics of pathology processes obtained from plasma of leukemia patients show that ATP value could be a prognostic factor of the treatment success. The role of ATP in the drug metabolism switching is also discussed within the context of kinetic modelling of metabolism processes leading to the production of 6-Thioguanosine monophosphate, which is a principal acting agent in chemotherapy.

  8. Adenosine-5'-triphosphate (ATP supplementation improves low peak muscle torque and torque fatigue during repeated high intensity exercise sets

    Directory of Open Access Journals (Sweden)

    Rathmacher John A

    2012-10-01

    Full Text Available Abstract Background Intracellular concentrations of adenosine-5’-triphosphate (ATP are many times greater than extracellular concentrations (1–10 mM versus 10–100 nM, respectively and cellular release of ATP is tightly controlled. Transient rises in extracellular ATP and its metabolite adenosine have important signaling roles; and acting through purinergic receptors, can increase blood flow and oxygenation of tissues; and act as neurotransmitters. Increased blood flow not only increases substrate availability but may also aid in recovery through removal of metabolic waste products allowing muscles to accomplish more work with less fatigue. The objective of the present study was to determine if supplemental ATP would improve muscle torque, power, work, or fatigue during repeated bouts of high intensity resistance exercise. Methods Sixteen participants (8 male and 8 female; ages: 21–34 years were enrolled in a double-blinded, placebo-controlled study using a crossover design. The participants received either supplemental ATP (400 mg/d divided into 2 daily doses or placebo for 15 d. After an overnight fast, participants underwent strength and fatigue testing, consisting of 3 sets of 50 maximal knee extensions performed on a Biodex® leg dynamometer. Results No differences were detected in high peak torque, power, or total work with ATP supplementation; however, low peak torque in set 2 was significantly improved (p Conclusions Supplementation with 400 mg ATP/d for 15 days tended to reduce muscle fatigue and improved a participant’s ability to maintain a higher force output at the end of an exhaustive exercise bout.

  9. (31)P-MRS of healthy human brain: ATP synthesis, metabolite concentrations, pH, and T1 relaxation times.

    Science.gov (United States)

    Ren, Jimin; Sherry, A Dean; Malloy, Craig R

    2015-11-01

    The conventional method for measuring brain ATP synthesis is (31)P saturation transfer (ST), a technique typically dependent on prolonged pre-saturation with γ-ATP. In this study, ATP synthesis rate in resting human brain is evaluated using EBIT (exchange kinetics by band inversion transfer), a technique based on slow recovery of γ-ATP magnetization in the absence of B1 field following co-inversion of PCr and ATP resonances with a short adiabatic pulse. The unidirectional rate constant for the Pi → γ-ATP reaction is 0.21 ± 0.04 s(-1) and the ATP synthesis rate is 9.9 ± 2.1 mmol min(-1)  kg(-1) in human brain (n = 12 subjects), consistent with the results by ST. Therefore, EBIT could be a useful alternative to ST in studying brain energy metabolism in normal physiology and under pathological conditions. In addition to ATP synthesis, all detectable (31)P signals are analyzed to determine the brain concentration of phosphorus metabolites, including UDPG at around 10 ppm, a previously reported resonance in liver tissues and now confirmed in human brain. Inversion recovery measurements indicate that UDPG, like its diphosphate analogue NAD, has apparent T1 shorter than that of monophosphates (Pi, PMEs, and PDEs) but longer than that of triphosphate ATP, highlighting the significance of the (31)P-(31)P dipolar mechanism in T1 relaxation of polyphosphates. Another interesting finding is the observation of approximately 40% shorter T1 for intracellular Pi relative to extracellular Pi, attributed to the modulation by the intracellular phosphoryl exchange reaction Pi ↔ γ-ATP. The sufficiently separated intra- and extracellular Pi signals also permit the distinction of pH between intra- and extracellular environments (pH 7.0 versus pH 7.4). In summary, quantitative (31)P MRS in combination with ATP synthesis, pH, and T1 relaxation measurements may offer a promising tool to detect biochemical alterations at early stages of brain dysfunctions and diseases.

  10. Students' Interdisciplinary Reasoning about "High-Energy Bonds" and ATP

    CERN Document Server

    Dreyfus, Benjamin W; Sawtelle, Vashti; Svoboda, Julia; Turpen, Chandra; Redish, Edward F

    2012-01-01

    Students' sometimes contradictory ideas about ATP (adenosine triphosphate) and the nature of chemical bonds have been studied in the biology and chemistry education literatures, but these topics are rarely part of the introductory physics curriculum. We present qualitative data from an introductory physics course for undergraduate biology majors that seeks to build greater interdisciplinary coherence and therefore includes these topics. In these data, students grapple with the apparent contradiction between the energy released when the phosphate bond in ATP is broken and the idea that an energy input is required to break a bond. We see that students' perceptions of how each scientific discipline bounds the system of interest can influence how they justify their reasoning about a topic that crosses disciplines. This has consequences for a vision of interdisciplinary education that respects disciplinary perspectives while bringing them into interaction in ways that demonstrate consistency amongst the perspectiv...

  11. Correlation of discocyte frequency and ATP concentration in preserved blood: A morphological indicator of red blood cell viability

    Directory of Open Access Journals (Sweden)

    Leonart M.S.S.

    1997-01-01

    Full Text Available Red blood cells (RBC are viable if kept in an adequate preservative solution, although gradual changes in morphology and metabolism may occur. There is a gradual decrease in adenosine-5'-triphosphate (ATP concentration, pH, glucose consumption, and enzyme activity during preservation. The normal discocyte shapes are initially replaced by echinocytes and stomatocytes and, at final stages, by spherocytes, the last step before splenic sequestration. Post-transfusional survival has been correlated with the ATP concentration. RBC preserved in ADSOL, a solution containing adenine, dextrose, sodium chloride, and mannitol, are viable for transfusion for up to 6 weeks. Erythrocytes from 10 blood units taken from healthy adult donors were preserved for 12 weeks in ADSOL at 4oC. We now report a significant correlation (r2 = 0.98 between the percentage of discocytes (89 to 7% and ATP (100 to 10% concentration in ADSOL-preserved RBC. The results suggest that the percent of discocyte shapes used as an indicator of ATP concentration may be a useful indicator for quality control of RBC viability in centers which have limited assay facilities.

  12. Fast monitoring of indoor bioaerosol concentrations with ATP bioluminescence assay using an electrostatic rod-type sampler.

    Directory of Open Access Journals (Sweden)

    Ji-Woon Park

    Full Text Available A culture-based colony counting method is the most widely used analytical technique for monitoring bioaerosols in both indoor and outdoor environments. However, this method requires several days for colony formation. In this study, our goal was fast monitoring (Sampling: 3 min, Detection: < 1 min of indoor bioaerosol concentrations with ATP bioluminescence assay using a bioaerosol sampler. For this purpose, a novel hand-held electrostatic rod-type sampler (110 mm wide, 115 mm long, and 200 mm tall was developed and used with a commercial luminometer, which employs the Adenosine triphosphate (ATP bioluminescence method. The sampler consisted of a wire-rod type charger and a cylindrical collector, and was operated with an applied voltage of 4.5 kV and a sampling flow rate of 150.7 lpm. Its performance was tested using Staphylococcus epidermidis which was aerosolized with an atomizer. Bioaerosol concentrations were measured using ATP bioluminescence method with our sampler and compared with the culture-based method using Andersen cascade impactor under controlled laboratory conditions. Indoor bioaerosol concentrations were also measured using both methods in various indoor environments. A linear correlation was obtained between both methods in lab-tests and field-tests. Our proposed sampler with ATP bioluminescence method may be effective for fast monitoring of indoor bioaerosol concentrations.

  13. High-altitude adaptation of Tibetan chicken from MT-COI and ATP-6 perspective.

    Science.gov (United States)

    Zhao, Xiaoling; Wu, Nan; Zhu, Qing; Gaur, Uma; Gu, Ting; Li, Diyan

    2016-09-01

    The problem of hypoxia adaptation in high altitudes is an unsolved brainteaser in the field of life sciences. As one of the best chicken breeds with adaptability to highland environment, the Tibetan chicken, is genetically different from lowland chicken breeds. In order to gain a better understanding of the mechanism of hypoxic adaptability in high altitude, in the present study, we focused on the MT-COI together with ATP-6 gene to explore the regulatory mechanisms for hypoxia adaptability in Tibet chicken. Here, we sequenced MT-COI of 29 Tibetan chickens and 30 Chinese domestic chickens and ATP-6 gene of 28 Tibetan chickens and 29 Chinese domestic chickens. In MT-COI gene, 9 single nucleotide polymorphisms (SNPs) were detected though none of these was a missense mutation, confirming the fact that MT-COI gene is a largely conservative sequence. In ATP-6 gene, 6 single nucleotide polymorphisms (SNPs) were detected and we found a missense mutation (m.9441G > A) in the ATP-6 gene of Tibetan chicken resulting in an amino acid substitution. Due to the critical role of ATP-6 gene in the proton translocation and energy metabolism, we speculated the possibility of this mutation playing an important role in easier energy conversion and metabolism in Tibetan chickens than Chinese domestic chickens so as to better adapt to the harsh environment of the high-altitude areas. The Median-joining profile also suggested that haplotype Ha2 has the ancestral position to the other haplotypes and has significant relationship with high-altitude adaptation in ATP-6 gene. Therefore, we considered that the polymorphism (m.9441G > A) in the ATP-6 gene may affect the specific functions of ATP-6 enzyme relating to high-altitude adaptation of Tibetan chicken and MT-COI gene is a largely conservative sequence.

  14. Integrating printed microfluidics with silicon photomultipliers for miniaturised and highly sensitive ATP bioluminescence detection.

    Science.gov (United States)

    Santangelo, M F; Libertino, S; Turner, A P F; Filippini, D; Mak, W C

    2018-01-15

    Bioluminescence has been widely used for important biosensing applications such as the measurement of adenosine triphosphate (ATP), the energy unit in biological systems and an indicator of vital processes. The current technology for detection is mainly based on large equipment such as readers and imaging systems, which require intensive and time-consuming procedures. A miniaturised bioluminescence sensing system, which would allow sensitive and continuous monitoring of ATP, with an integrated and low-cost disposable microfluidic chamber for handling of biological samples, is highly desirable. Here, we report the design, fabrication and testing of 3D printed microfluidics chips coupled with silicon photomultipliers (SiPMs) for high sensitive real-time ATP detection. The 3D microfluidic chip reduces reactant consumption and facilitates solution delivery close to the SiPM to increase the detection efficiency. Our system detects ATP with a limit of detection (LoD) of 8nM and an analytical dynamic range between 15nM and 1µM, showing a stability error of 3%, and a reproducibility error below of 20%. We demonstrate the dynamic monitoring of ATP in a continuous-flow system exhibiting a fast response time, ~4s, and a full recovery to the baseline level within 17s. Moreover, the SiPM-based bioluminescence sensing system shows a similar analytical dynamic range for ATP detection to that of a full-size PerkinElmer laboratory luminescence reader. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Fast monitoring of indoor bioaerosol concentrations with ATP bioluminescence assay using an electrostatic rod-type sampler.

    Science.gov (United States)

    Park, Ji-Woon; Park, Chul Woo; Lee, Sung Hwa; Hwang, Jungho

    2015-01-01

    A culture-based colony counting method is the most widely used analytical technique for monitoring bioaerosols in both indoor and outdoor environments. However, this method requires several days for colony formation. In this study, our goal was fast monitoring (Sampling: 3 min, Detection: bioluminescence assay using a bioaerosol sampler. For this purpose, a novel hand-held electrostatic rod-type sampler (110 mm wide, 115 mm long, and 200 mm tall) was developed and used with a commercial luminometer, which employs the Adenosine triphosphate (ATP) bioluminescence method. The sampler consisted of a wire-rod type charger and a cylindrical collector, and was operated with an applied voltage of 4.5 kV and a sampling flow rate of 150.7 lpm. Its performance was tested using Staphylococcus epidermidis which was aerosolized with an atomizer. Bioaerosol concentrations were measured using ATP bioluminescence method with our sampler and compared with the culture-based method using Andersen cascade impactor under controlled laboratory conditions. Indoor bioaerosol concentrations were also measured using both methods in various indoor environments. A linear correlation was obtained between both methods in lab-tests and field-tests. Our proposed sampler with ATP bioluminescence method may be effective for fast monitoring of indoor bioaerosol concentrations.

  16. Release of ATP from marginal cells in the cochlea of neonatal rats can be induced by changes in extracellular and intracellular ion concentrations.

    Directory of Open Access Journals (Sweden)

    Yating Peng

    Full Text Available BACKGROUND: Adenosine triphosphate (ATP plays an important role in the cochlea. However, the source of ATP and the mechanism by which it is released remain unclear. This study investigates the presence and release mechanism of ATP in vitro cultured marginal cells isolated from the stria vascularis of the cochlea in neonatal rats. METHODS: Sprague-Dawley rats aged 1-3 days old were used for isolation, in vitro culture, and purification of marginal cells. Cultured marginal cells were verified by flow cytometry. Vesicles containing ATP in these cells were identified by fluorescence staining. The bioluminescence assay was used for determination of ATP concentration in the extracellular fluid released by marginal cells. Assays for ATP concentration were performed when the ATP metabolism of cells was influenced, and ionic concentrations in intracellular and extracellular fluid were found to change. RESULTS: Evaluation of cultured marginal cells with flow cytometry revealed the percentage of fluorescently-labeled cells as 92.9% and 81.9%, for cytokeratin and vimentin, respectively. Quinacrine staining under fluorescence microscopy revealed numerous green, star-like spots in the cytoplasm of these cells. The release of ATP from marginal cells was influenced by changes in the concentration of intracellular and extracellular ions, namely extracellular K(+ and intra- and extracellular Ca(2+. Furthermore, changes in the concentration of intracellular Ca(2+ induced by the inhibition of the phospholipase signaling pathway also influence the release of ATP from marginal cells. CONCLUSION: We confirmed the presence and release of ATP from marginal cells of the stria vascularis. This is the first study to demonstrate that the release of ATP from such cells is associated with the state of the calcium pump, K(+ channel, and activity of enzymes related to the phosphoinositide signaling pathway, such as adenylate cyclase, phospholipase C, and phospholipase A(2.

  17. Glucose-stimulated oscillations in free cytosolic ATP concentration imaged in single islet beta-cells: evidence for a Ca2+-dependent mechanism.

    Science.gov (United States)

    Ainscow, Edward K; Rutter, Guy A

    2002-02-01

    Normal glucose-stimulated insulin secretion is pulsatile, but the molecular mechanisms underlying this pulsatility are poorly understood. Oscillations in the intracellular free [ATP]/[ADP] ratio represent one possible mechanism because they would be expected to cause fluctuations in ATP-sensitive K(+) channel activity and hence oscillatory Ca(2+) influx. After imaging recombinant firefly luciferase, expressed via an adenoviral vector in single human or mouse islet beta-cells, we report here that cytosolic free ATP concentrations oscillate and that these oscillations are affected by glucose. In human beta-cells, oscillations were observed at both 3 and 15 mmol/l glucose, but the oscillations were of a longer wavelength at the higher glucose concentration (167 vs. 66 s). Mouse beta-cells displayed oscillations in both cytosolic free [Ca(2+)] and [ATP] only at elevated glucose concentrations, both with a period of 120 s. To explore the causal relationship between [Ca(2+)] and [ATP] oscillations, the regulation of each was further investigated in populations of MIN6 beta-cells. Incubation in Ca(2+)-free medium lowered cytosolic [Ca(2+)] but increased [ATP] in MIN6 cells at both 3 and 30 mmol/l glucose. Removal of external Ca(2+) increased [ATP], possibly by decreasing ATP consumption by endoplasmic reticulum Ca(2+)-ATPases. These results allow a model to be constructed of the beta-cell metabolic oscillator that drives nutrient-induced insulin secretion.

  18. Calcium regulates motility and protein phosphorylation by changing cAMP and ATP concentrations in boar sperm in vitro.

    Science.gov (United States)

    Li, Xinhong; Wang, Lirui; Li, Yuhua; Zhao, Na; Zhen, Linqing; Fu, Jieli; Yang, Qiangzhen

    2016-09-01

    Considering the importance of calcium (Ca(2+)) in regulating sperm capacitation, hyperactivation and acrosome reaction, little is known about the molecular mechanism of action of this ion in this process. In the present study, assessment of the molecular mechanism from the perspective of energy metabolism occurred. Sperm motility variables were determined using computer-assisted sperm analysis (CASA) and the phosphorylation of PKA substrates, tyrosine residues and AMP-activated protein kinase (AMPK) were analyzed by Western blot. Moreover, intracellular sperm-specific glyceraldehyde 3-phosphatedehydrogenase (GAPDH) activity, 3'-5'-cyclic adenosine monophosphate (cAMP) and adenosine 5'-triphosphate (ATP) concentrations were assessed in boar sperm treated with Ca(2+). Results of the present study indicated that, under greater extracellular Ca(2+)concentrations (≥3.0mM), sperm motility and protein phosphorylation were inhibited. Interestingly, these changes were correlated with that of GAPDH activity, AMPK phosphorylation, cAMP and ATP concentrations. The negative effects of Ca(2+) on these intracellular processes were attenuated by addition of the calmodulin (CaM) inhibitor W7 and the inhibitor of calmodulin-dependent protein kinase (CaMK), KN-93. In the presence of greater extracellular Ca(2+), however, the phosphorylation pathway was suppressed by H-89. Taken together, these results suggested that Ca(2+) had a dual role in regulating boar sperm motility and protein phosphorylation due to the changes of cAMP and ATP concentrations, in response to cAMP-mediated signal transduction and the Ca(2+) signaling cascade. The present study provided some novel insights into the molecular mechanism underlying the effects of Ca(2+) on boar sperm as well as the involvement of energy metabolism in this mechanism.

  19. Impacts of high ATP supply from chloroplasts and mitochondria on the leaf metabolism of Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Chao eLiang

    2015-10-01

    Full Text Available Chloroplasts and mitochondria are the major ATP producing organelles in plant leaves. Arabidopsis thaliana purple acid phosphatase 2 (AtPAP2 is a phosphatase dually targeted to the outer membranes of both organelles and it plays a role in the import of selected nuclear-encoded proteins into these two organelles. Overexpression (OE of AtPAP2 in Arabidopsis thaliana accelerates plant growth and promotes flowering, seed yield and biomass at maturity. Measurement of ADP/ATP/NADP+/NADPH contents in the leaves of 20-day-old OE and wild-type lines at the end of night and at 1 and 8 h following illumination in a 16/8 h photoperiod revealed that the ATP levels and ATP/NADPH ratios were significantly increased in the OE line at all three time points. The AtPAP2 OE line is therefore a good model to investigate the impact of high energy on the global molecular status of Arabidopsis. In this study, transcriptome, proteome and metabolome profiles of the high ATP transgenic line were examined and compared with those of wild-type plants. A comparison of OE and WT at the end of the night provide valuable information on the impact of higher ATP output from mitochondria on plant physiology, as mitochondrial respiration is the major source of ATP in the dark in leaves. Similarly, comparison of OE and WT following illumination will provide information on the impact of higher energy output from chloroplasts on plant physiology. Overexpression of AtPAP2 was found to significantly affect the transcript and protein abundances of genes encoded by the two organellar genomes. For example, the protein abundances of many ribosomal proteins encoded by the chloroplast genome were higher in the AtPAP2 OE line under both light and dark conditions, while the protein abundances of multiple components of the photosynthetic complexes were lower. RNA-seq data also showed that the transcription of the mitochondrial genome is greatly affected by the availability of energy. These data

  20. Effect of extracellular adenosine 5'-triphosphate on cryopreserved epididymal cat sperm intracellular ATP concentration, sperm quality, and in vitro fertilizing ability.

    Science.gov (United States)

    Thuwanut, Paweena; Arya, Nlin; Comizzoli, Pierre; Chatdarong, Kaywalee

    2015-09-15

    Intracellular adenosine 5'-triphosphate (ATP) is essential for supporting sperm function in the fertilization process. During cryopreservation, damage of sperm mitochondrial membrane usually leads to compromised production of intracellular ATP. Recently, extracellular ATP (ATPe) was introduced as a potent activator of sperm motility and fertilizing ability. This study aimed to evaluate (1) levels of intracellular ATP in frozen-thawed epididymal cat sperm after incubation with ATPe and (2) effects of ATPe on epididymal cat sperm parameters after freezing and thawing. Eighteen male cats were included. For each replicate, epididymal sperm from two cats were pooled to one sample (N = 9). Each pooled sample was cryopreserved with the Tris-egg yolk extender into three straws. After thawing, the first and second straws were incubated with 0-, 1.0-, or 2.5-mM ATPe for 10 minutes and evaluated for sperm quality at 10 minutes, 1, 3, and 6 hours after thawing and fertilizing ability. The third straw was evaluated for intracellular ATP concentration in control and with 2.5-mM ATPe treatment. Higher concentration of intracellular sperm ATP was observed in the samples treated with 2.5-mM ATPe compared to the controls (0.339 ± 0.06 μg/2 × 10(6) sperm vs. 0.002 ± 0.003 μg/2 × 10(6) sperm, P ≤ 0.05). In addition, incubation with 2.5-mM ATPe for 10 minutes promoted sperm motility (56.7 ± 5.0 vs. 53.3 ± 4.4%, P ≤ 0.05) and progressive motility (3.1 ± 0.2 vs. 2.8 ± 0.4, P ≤ 0.05), mitochondrial membrane potential (36.4 ± 5.5 vs. 28.7 ± 4.8%, P ≤ 0.05), and blastocyst rate (36.1 ± 7.0 and 28.8 ± 7.4%, P ≤ 0.05) compared with the controls. In contrast, ATPe remarkably interfered acrosome integrity after 6 hours of postthawed incubation. In sum, the present finding that optimal incubation time of postthaw epididymal cat sperm under proper ATPe condition might constitute a rationale for the studies on other endangered wild felids regarding sperm quality and embryo

  1. A simple high-content cell cycle assay reveals frequent discrepancies between cell number and ATP and MTS proliferation assays.

    Directory of Open Access Journals (Sweden)

    Grace Ka Yan Chan

    Full Text Available In order to efficiently characterize both antiproliferative potency and mechanism of action of small molecules targeting the cell cycle, we developed a high-throughput image-based assay to determine cell number and cell cycle phase distribution. Using this we profiled the effects of experimental and approved anti-cancer agents with a range mechanisms of action on a set of cell lines, comparing direct cell counting versus two metabolism-based cell viability/proliferation assay formats, ATP-dependent bioluminescence, MTS (3-(4,5-dimethylthiazol-2-yl-5-(3-carboxymethoxyphenyl-2-(4-sulfophenyl-2H-tetrazolium reduction, and a whole-well DNA-binding dye fluorescence assay. We show that, depending on compound mechanisms of action, the metabolism-based proxy assays are frequently prone to 1 significant underestimation of compound potency and efficacy, and 2 non-monotonic dose-response curves due to concentration-dependent phenotypic 'switching'. In particular, potency and efficacy of DNA synthesis-targeting agents such as gemcitabine and etoposide could be profoundly underestimated by ATP and MTS-reduction assays. In the same image-based assay we showed that drug-induced increases in ATP content were associated with increased cell size and proportionate increases in mitochondrial content and respiratory flux concomitant with cell cycle arrest. Therefore, differences in compound mechanism of action and cell line-specific responses can yield significantly misleading results when using ATP or tetrazolium-reduction assays as a proxy for cell number when screening compounds for antiproliferative activity or profiling panels of cell lines for drug sensitivity.

  2. High Sensitivity Detection of ATP Using Bioluminescence at An Optical Fiber End

    Science.gov (United States)

    Iinuma, Masataka; Ushio, Yasuaki; Kuroda, Akio; Kadoya, Yutaka

    We investigated the sensitivity of ATP detection based on bioluminescence at an optical fiber end where luciferase molecules were immobilized via silica-binding protein molecules. Luminescence was detected by an avalanche photo diode (APD), with coupling optics to make full use of the merit of compactness, high quantum efficiency and low noise of the APD. The core diameter and the numerical aperture of the optical fiber, as well as the design of the coupling optics, were optimized so as to realize high photon-collection efficiency. A detection limit of about 10-10 M was obtained, which corresponds to 10-15 mol of ATP. A rough estimation shows that the photon count rate is still two orders of magnitude lower than that limited by diffusion or reaction processes, implying a possibility of further improvement of the sensitivity.

  3. The crustacean gill (Na+,K+)-ATPase: allosteric modulation of high- and low-affinity ATP-binding sites by sodium and potassium.

    Science.gov (United States)

    Masui, D C; Silva, E C C; Mantelatto, F L M; McNamara, J C; Barrabin, H; Scofano, H M; Fontes, C F L; Furriel, R P M; Leone, F A

    2008-11-15

    The blue crab, Callinectes danae, tolerates exposure to a wide salinity range employing mechanisms of compensatory ion uptake when in dilute media. Although the gill (Na+,K+)-ATPase is vital to hyperosmoregulatory ability, the interactions occurring at the sites of ATP binding on the molecule itself are unknown. Here, we investigate the modulation by Na+ and K+ of homotropic interactions between the ATP-binding sites, and of phosphoenzyme formation of the (Na+,K+)-ATPase from the posterior gills of this euryhaline crab. The contribution of the high- and low-affinity ATP-binding sites to maximum velocity was similar for both Na+ and K+. However, in contrast to Na+, a threshold K+ concentration triggers the appearance of the high-affinity binding sites, displacing the saturation curve to lower ATP concentrations.Further, a low-affinity site for phosphorylation is present on the enzyme. These findings reveal notable differences in the catalytic mechanism of the crustacean (Na+,K+)-ATPase compared to the vertebrate enzyme.

  4. A possible role for extra-cellular ATP in plant responses to high frequency, low amplitude electromagnetic field.

    Science.gov (United States)

    Roux, David; Faure, Catherine; Bonnet, Pierre; Girard, Sébastien; Ledoigt, Gérard; Davies, Eric; Gendraud, Michel; Paladian, Françoise; Vian, Alain

    2008-06-01

    In parallel to evoking the accumulation of stress-related transcripts, exposure to low level 900 MHz EMF affected the levels of ATP, the main energy molecule of the cell. Its concentration dropped rapidly (27% after 30 min) in response to EMF exposure, along with a 18% decrease in the adenylate energy charge (AEC), a good marker of cell energy status. One could interpret this decrease in ATP and AEC in a classical way, i.e., as the result of an increase in cellular energy usage, but recent work brings exciting new insights in pointing out a signalling function for ATP, especially in the stress physiology context where it could trigger both reactive oxygen species and calcium movement (this latter being involved in plant responses to EMF exposure). In this addendum, we discuss our results within this new perspective for ATP function.

  5. Dose-dependent ATP depletion and cancer cell death following calcium electroporation, relative effect of calcium concentration and electric field strength

    DEFF Research Database (Denmark)

    Hansen, Emilie Louise; Sozer, Esin Bengisu; Romeo, Stefania;

    2015-01-01

    death and could be a novel cancer treatment. This study aims at understanding the relationship between applied electric field, calcium concentration, ATP depletion and efficacy. METHODS: In three human cell lines--H69 (small-cell lung cancer), SW780 (bladder cancer), and U937 (leukaemia), viability...... was observed with fluorescence confocal microscopy of quinacrine-labelled U937 cells. RESULTS: Both H69 and SW780 cells showed dose-dependent (calcium concentration and electric field) decrease in intracellular ATP (p...-dependently reduced cell survival and intracellular ATP. Increasing extracellular calcium allows the use of a lower electric field. GENERAL SIGNIFICANCE: This study supports the use of calcium electroporation for treatment of cancer and possibly lowering the applied electric field in future trials....

  6. Dead-end ultrafiltration concentration and IMS/ATP-bioluminescence detection of Escherichia coli O157:H7 in recreational water and produce wash.

    Science.gov (United States)

    Hunter, Dawn M; Leskinen, Stephaney D; Magaña, Sonia; Schlemmer, Sarah M; Lim, Daniel V

    2011-12-01

    The purpose of this study was to develop a detection method for viable E. coli O157:H7 in fresh produce and recreational water. The method was evaluated using eight samples of produce wash and recreational water with or without spiked E. coli O157:H7 at ≤10(2) CFU·ml(-1) and concentrated using dead-end ultrafiltration (DEUF) to produce primary and secondary retentates. Fifty-four matrix replicates of undiluted secondary retentates or dilutions (1:2 or 1:10 in buffer) were evaluated using an IMS/ATP bioluminescence assay (IMS/ATP). Combining primary and secondary DEUF yielded a 2-4 log(10) increase in E. coli O157:H7 concentrations in spiked samples and resulted in signal-to-noise ratios 2-219 fold higher than controls, depending on the sample type. DEUF increased the concentration of E. coli O157:H7 to within the detectable limits of IMS/ATP. The combined assay provided detection of viable E. coli O157:H7 in produce and recreational water. Accurate detection of microbial pathogens using DEUF and IMS/ATP could reduce disease outbreaks from contaminated water sources and food products.

  7. ATP signals

    DEFF Research Database (Denmark)

    Novak, Ivana

    2016-01-01

    The Department of Biology at the University of Copenhagen explains the function of ATP signalling in the pancreas......The Department of Biology at the University of Copenhagen explains the function of ATP signalling in the pancreas...

  8. A receptor that is highly specific for extracellular ATP in developing chick skeletal muscle in vitro.

    OpenAIRE

    Thomas, S A; Zawisa, M. J.; X. Lin; Hume, R. I.

    1991-01-01

    1. Extracellular adenosine 5'-triphosphate (ATP) activated an early excitatory conductance followed by a late potassium conductance in developing chick skeletal muscle. A series of ATP analogues were tested for their ability to activate these two conductances. All compounds tested were either agonists for both responses or for neither. Furthermore, the potency of agonists was similar for the two responses. 2. The order of potency for agonists was ATP approximately adenosine 5'-O-(3-thiotripho...

  9. Highly Dynamic Interactions Maintain Kinetic Stability of the ClpXP Protease During the ATP-Fueled Mechanical Cycle.

    Science.gov (United States)

    Amor, Alvaro J; Schmitz, Karl R; Sello, Jason K; Baker, Tania A; Sauer, Robert T

    2016-06-17

    The ClpXP protease assembles in a reaction in which an ATP-bound ring hexamer of ClpX binds to one or both heptameric rings of the ClpP peptidase. Contacts between ClpX IGF-loops and clefts on a ClpP ring stabilize the complex. How ClpXP stability is maintained during the ATP-hydrolysis cycle that powers mechanical unfolding and translocation of protein substrates is poorly understood. Here, we use a real-time kinetic assay to monitor the effects of nucleotides on the assembly and disassembly of ClpXP. When ATP is present, complexes containing single-chain ClpX assemble via an intermediate and remain intact until transferred into buffers containing ADP or no nucleotides. ATP binding to high-affinity subunits of the ClpX hexamer prevents rapid dissociation, but additional subunits must be occupied to promote assembly. Small-molecule acyldepsipeptides, which compete with the IGF loops of ClpX for ClpP-cleft binding, cause exceptionally rapid dissociation of otherwise stable ClpXP complexes, suggesting that the IGF-loop interactions with ClpP must be highly dynamic. Our results indicate that the ClpX hexamer spends almost no time in an ATP-free state during the ATPase cycle, allowing highly processive degradation of protein substrates.

  10. The High Altitude Balloon Experiment demonstration of acquisition, tracking, and pointing technologies (HABE-ATP)

    Science.gov (United States)

    Dimiduk, D.; Caylor, M.; Williamson, D.; Larson, L.

    1995-01-01

    The High Altitude Balloon Experiment demonstration of Acquisition, Tracking, and Pointing (HABE-ATP) is a system built around balloon-borne payload which is carried to a nominal 26-km altitude. The goal is laser tracking thrusting theater and strategic missiles, and then pointing a surrogate laser weapon beam, with performance levels end a timeline traceable to operational laser weapon system requirements. This goal leads to an experiment system design which combines hardware from many technology areas: an optical telescope and IR sensors; an advanced angular inertial reference; a flexible multi-level of actuation digital control system; digital tracking processors which incorporate real-time image analysis and a pulsed, diode-pumped solid state tracking laser. The system components have been selected to meet the overall experiment goals of tracking unmodified boosters at 50- 200 km range. The ATP system on HABE must stabilize and control a relative line of sight between the platform and the unmodified target booster to a 1 microrad accuracy. The angular pointing reference system supports both open loop and closed loop track modes; GPS provides absolute position reference. The control system which positions the line of sight for the ATP system must sequence through accepting a state vector handoff, closed-loop passive IR acquisition, passive IR intermediate fine track, active fine track, and then finally aimpoint determination and maintenance modes. Line of sight stabilization to fine accuracy levels is accomplished by actuating wide bandwidth fast steering mirrors (FSM's). These control loops off-load large-amplitude errors to the outer gimbal in order to remain within the limited angular throw of the FSM's. The SWIR acquisition and MWIR intermediate fine track sensors (both PtSi focal planes) image the signature of the rocket plume. After Hard Body Handover (HBHO), active fine tracking is conducted with a visible focal plane viewing the laser-illuminated target

  11. High-efficiency solar concentrator

    Science.gov (United States)

    Lansing, F. L.; Dorman, J.

    1980-01-01

    A new type of solar concentrator is presented using liquid lenses and simple translational tracking mechanism. The concentrator achieves a 100:1 nominal concentration ratio and is compared in performance with a flat-plate collector having two sheets of glazing and non-selective coating. The results of the thermal analysis show that higher temperatures can be obtained with the concentrator than is possible with the non-concentrator flat-plate type. Furthermore, the thermal efficiency far exceeds that of the comparative flat-plate type for all operating conditions.

  12. [Evaluation of renal damage using urinary ATP analysis].

    Science.gov (United States)

    Uehara, Yuki; Yanai, Mitsuru; Kumasaka, Kazunari

    2004-10-01

    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.

  13. Sleep, brain energy levels, and food intake: Relationship between hypothalamic ATP concentrations, food intake, and body weight during sleep-wake and sleep deprivation in rats.

    Science.gov (United States)

    Dworak, M; Kim, T; McCarley, R W; Basheer, R

    2011-06-01

    The feeling of hunger and feeding, a wake-state-dependent behavior, is regulated by specific centers within the hypothalamus. While paraventricular nucleus (PVN), arcuate nucleus (ARC), and dorso- and ventromedial hypothalamus (DMH/VMH) regulate feeding, the lateral hypothalamus (LH) is associated both with feeding and wake/REM sleep regulation. In order to examine the effects of sleep and wakefulness on food intake and body weight, we also measured hypothalamic ATP concentrations, which are known to be involved in feeding behavior and sleep-wake regulation. In rats, food intake and body weight was measured during a 24-h light-dark cycle and during 6 h of sleep deprivation (SD) performed by gentle handling. Tissue samples from the PVN, ARC/DMH/VMH, and LH were collected after 6 h of SD and from time-matched diurnal controls. ATP was measured by luciferin-luciferase bioluminescence assay. Across the 24-h light-dark period, rats consumed approximately 28.13±4.48 g of food and gained 5.22±1.65 g with a positive correlation between food intake and body weight. During SD, while food intake increased significantly +147.31±6.13%, they lost weight significantly (-93.29±13.64%) when compared to undisturbed controls. SD resulted in a significant decrease in ATP levels only in LH (-44.60±21.13%) with no change in PVN, ARC/DMH/VMH region when compared with undisturbed controls. The results indicate a strong overall correlation between ATP concentrations in the LH and individual food intake and suggest a sleep-wake dependent neuronal control of food intake and body weight.

  14. Glucose generates sub-plasma membrane ATP microdomains in single islet beta-cells. Potential role for strategically located mitochondria.

    Science.gov (United States)

    Kennedy, H J; Pouli, A E; Ainscow, E K; Jouaville, L S; Rizzuto, R; Rutter, G A

    1999-05-01

    Increases in the concentration of free ATP within the islet beta-cell may couple elevations in blood glucose to insulin release by closing ATP-sensitive K+ (KATP) channels and activating Ca2+ influx. Here, we use recombinant targeted luciferases and photon counting imaging to monitor changes in free [ATP] in subdomains of single living MIN6 and primary beta-cells. Resting [ATP] in the cytosol ([ATP]c), in the mitochondrial matrix ([ATP]m), and beneath the plasma membrane ([ATP]pm) were similar ( approximately 1 mM). Elevations in extracellular glucose concentration (3-30 mM) increased free [ATP] in each domain with distinct kinetics. Thus, sustained increases in [ATP]m and [ATP]pm were observed, but only a transient increase in [ATP]c. However, detectable increases in [ATP]c and [ATP]pm, but not [ATP]m, required extracellular Ca2+. Enhancement of glucose-induced Ca2+ influx with high [K+] had little effect on the apparent [ATP]c and [ATP]m increases but augmented the [ATP]pm increase. Underlying these changes, glucose increased the mitochondrial proton motive force, an effect mimicked by high [K+]. These data support a model in which glucose increases [ATP]m both through enhanced substrate supply and by progressive Ca2+-dependent activation of mitochondrial enzymes. This may then lead to a privileged elevation of [ATP]pm, which may be essential for the sustained closure of KATP channels. Luciferase imaging would appear to be a useful new tool for dynamic in vivo imaging of free ATP concentration.

  15. Quantitative autoradiography of the binding sites for ( sup 125 I) iodoglyburide, a novel high-affinity ligand for ATP-sensitive potassium channels in rat brain

    Energy Technology Data Exchange (ETDEWEB)

    Gehlert, D.R.; Gackenheimer, S.L.; Mais, D.E.; Robertson, D.W. (Eli Lilly and Co., Indianapolis, IN (USA))

    1991-05-01

    We have developed a high specific activity ligand for localization of ATP-sensitive potassium channels in the brain. When brain sections were incubated with ({sup 125}I)iodoglyburide (N-(2-((((cyclohexylamino)carbonyl)amino)sulfonyl)ethyl)-5-{sup 125}I-2- methoxybenzamide), the ligand bound to a single site with a KD of 495 pM and a maximum binding site density of 176 fmol/mg of tissue. Glyburide was the most potent inhibitor of specific ({sup 125}I)iodoglyburide binding to rat forebrain sections whereas iodoglyburide and glipizide were slightly less potent. The binding was also sensitive to ATP which completely inhibited binding at concentrations of 10 mM. Autoradiographic localization of ({sup 125}I)iodoglyburide binding indicated a broad distribution of the ATP-sensitive potassium channel in the brain. The highest levels of binding were seen in the globus pallidus and ventral pallidum followed by the septohippocampal nucleus, anterior pituitary, the CA2 and CA3 region of the hippocampus, ventral pallidum, the molecular layer of the cerebellum and substantia nigra zona reticulata. The hilus and dorsal subiculum of the hippocampus, molecular layer of the dentate gyrus, cerebral cortex, lateral olfactory tract nucleus, olfactory tubercle and the zona incerta contained relatively high levels of binding. A lower level of binding (approximately 3- to 4-fold) was found throughout the remainder of the brain. These results indicate that the ATP-sensitive potassium channel has a broad presence in the rat brain and that a few select brain regions are enriched in this subtype of neuronal potassium channels.

  16. Mesurements of intracellular ATP provide new insight into the regulation of glycolysis in the yeast Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Ytting, Cecilie Karkov; Fuglsang, Anja Thoe; Hiltunen, J. Kalervo

    2012-01-01

    Glycolysis in the yeast Saccharomyces cerevisiae exhibits temporal oscillation under anaerobic or semianaerobic conditions. Previous evidence indicated that at least two membrane-bound ATPases, the mitochondrial F0F1 ATPase and the plasma membrane P-type ATPase (Pma1p), were important in regulating...... of the temporal behaviour of intracellular ATP in a yeast strain with oscillating glycolysis showed that, in addition to oscillation in intracellular ATP, there is an overall slow decrease in intracellular ATP because the ATP consumption rate exceeds the ATP production in glycolysis. Measurements of the temporal...... activity is under strict control. In the absence of glucose ATPase activity is switched off, and the intracellular ATP concentration is high. When glucose is added to the cells the ATP concentration starts to decrease, because ATP consumption exceeds ATP production by glycolysis. Finally, when glucose...

  17. Fabricating high-energy quantum dots in ultra-thin LiFePO4 nanosheets using a multifunctional high-energy biomolecule-ATP

    DEFF Research Database (Denmark)

    Zhang, X.D.; Bi, Z.Y.; He, W.

    2014-01-01

    By using a multifunctional high-energy biomolecule—adenosine triphosphate (ATP)—we fabricated highenergy quantum dots (HEQDs) with a feature size of less than 10 nm and used them in high-power lithium-ion batteries. We introduced high-energy phosphate bonds into the crystal structure of LiFePO4...

  18. Design and development of a high-concentration photovoltaic concentrator

    Energy Technology Data Exchange (ETDEWEB)

    Hodge, R C

    1982-04-01

    The design and development of a high concentration photovoltaic concentrator module is discussed. The design concept described herein incorporates a curved groove domed Fresnel lens, a high concentration etched multiple vertical junction (EMVJ) solar cell and a passively cooled direct-bonded copper cell mount all packaged in a plastic module. Two seven inch diameter 1200x domed Fresnel lenses were fabricated using single point diamond turning technology. Testing at both GE and Sandia confirmed optical transmission efficiencies of over 83%. Samples of the latest available EMVJ cells were mounted and installed, with a domed Fresnel lens, into a prototype module. Subsequent testing demonstrated net lens-cell efficiencies of 10 to 13%. As a result of this program, salient conclusions have been formulated as to this technology.

  19. A Vision of Interdisciplinary Education: Students' Reasoning about "High-Energy Bonds" and ATP

    CERN Document Server

    Dreyfus, Benjamin W; Turpen, Chandra; Gouvea, Julia; Redish, Edward F

    2014-01-01

    As interdisciplinary courses are developed, instructors and researchers have to grapple with questions of how students should make connections across disciplines. We explore the issue of interdisciplinary reconciliation (IDR): how students reconcile seemingly contradictory ideas from different disciplines. While IDR has elements in common with other frameworks for the reconciliation of ideas across contexts, it differs in that each disciplinary idea is considered canonically correct within its own discipline. The setting for the research is an introductory physics course for biology majors that seeks to build greater interdisciplinary coherence and therefore includes biologically relevant topics such as ATP and chemical bond energy. In our case-study data, students grapple with the apparent contradiction between the energy released when the phosphate bond in ATP is broken and the idea that an energy input is required to break a bond. We see students justifying context-dependent modeling choices, showing nuanc...

  20. Stimulation of primary osteoblasts with ATP induces transient vinculin clustering at sites of high intracellular traction force.

    Science.gov (United States)

    Tan, Toh Weng; Pfau, Bastian; Jones, David; Meyer, Thomas

    2014-02-01

    Adenosine 5'-triphosphate (ATP), released in response to mechanical and inflammatory stimuli, induces the dynamic and asynchronous protrusion and subsequent retraction of local membrane structures in osteoblasts. The molecular mechanisms involved in the ligand-stimulated herniation of the plasma membrane are largely unknown, which prompted us to investigate whether the focal-adhesion protein vinculin is engaged in the cytoskeletal alterations that underlie the ATP-induced membrane blebbing. Using time-lapse fluorescence microscopy of primary bovine osteoblast-like cells expressing green fluorescent protein-tagged vinculin, we found that stimulation of cells with 100 μM ATP resulted in the transient and rapid clustering of recombinant vinculin in the cell periphery, starting approximately 100 s after addition of the nucleotide. The ephemeral nature of the vinculin clusters was made evident by the brevity of their mean assembly and disassembly times (66.7 ± 13.3 s and 99.0 ± 6.6 s, respectively). Traction force vector maps demonstrated that the vinculin-rich clusters were localized predominantly at sites of high traction force. Intracellular calcium measurements showed that the ligand-induced increase in [Ca(2+)]i clearly preceded the clustering of vinculin, since [Ca(2+)]i levels returned to normal within 30 s of exposure to ATP, indicating that intracellular calcium transients trigger a cascade of signalling events that ultimately result in the incorporation of vinculin into membrane-associated focal aggregates.

  1. Behavior and stability of adenosine triphosphate (ATP) during chlorine disinfection.

    Science.gov (United States)

    Nescerecka, Alina; Juhna, Talis; Hammes, Frederik

    2016-09-15

    Adenosine triphosphate (ATP) analysis is a cultivation-independent alternative method for the determination of bacterial viability in both chlorinated and non-chlorinated water. Here we investigated the behavior and stability of ATP during chlorination in detail. Different sodium hypochlorite doses (0-22.4 mg-Cl2 L(-1); 5 min exposure) were applied to an Escherichia coli pure culture suspended in filtered river water. We observed decreasing intracellular ATP with increasing chlorine concentrations, but extracellular ATP concentrations only increased when the chlorine dose exceeded 0.35 mg L(-1). The release of ATP from chlorine-damaged bacteria coincided with severe membrane damage detected with flow cytometry (FCM). The stability of extracellular ATP was subsequently studied in different water matrixes, and we found that extracellular ATP was stable in sterile deionized water and also in chlorinated water until extremely high chlorine doses (≤11.2 mg-Cl2 L(-1); 5 min exposure). In contrast, ATP decreased relatively slowly (k = 0.145 h(-1)) in 0.1 μm filtered river water, presumably due to degradation by either extracellular enzymes or the fraction of bacteria that were able to pass through the filter. Extracellular ATP decreased considerably faster (k = 0.368 h(-1)) during batch growth of a river water bacterial community. A series of growth potential tests showed that extracellular ATP molecules were utilized as a phosphorus source during bacteria proliferation. From the combined data we conclude that ATP released from bacteria at high chlorine doses could promote bacteria regrowth, contributing to biological instability in drinking water distribution systems.

  2. High-resolution structure of the rotor ring of a proton-dependent ATP synthase.

    Science.gov (United States)

    Pogoryelov, Denys; Yildiz, Ozkan; Faraldo-Gómez, José D; Meier, Thomas

    2009-10-01

    The crystal structure of the c-ring from the proton-coupled F1Fo ATP synthase from Spirulina platensis is shown at 2.1-A resolution. The ring includes 15 membrane-embedded c subunits forming an hourglass-shaped assembly. The structure demonstrates that proton translocation across the membrane entails protonation of a conserved glutamate located near the membrane center in the c subunit outer helix. The proton is locked in this site by a precise hydrogen bond network reminiscent of that in Na+-dependent ATP synthases. However, the structure suggests that the different coordination chemistry of the bound proton and the smaller curvature of the outer helix drastically enhance the selectivity of the H+ site against other cations, including H3O+. We propose a model for proton translocation whereby the c subunits remain in this proton-locked state when facing the membrane lipid. Proton exchange would occur in a more hydrophilic and electrostatically distinct environment upon contact with the a subunit interface.

  3. Adenosine 5 '-triphosphate (ATP) supplements are not orally bioavailable: a randomized, placebo-controlled cross-over trial in healthy humans

    NARCIS (Netherlands)

    Arts, I.C.W.; Coolen, E.J.C.M.; Bours, M.J.L.; Huyghebaert, N.; Cohen Stuart, M.A.; Bast, A.; Dagnelie, P.C.

    2012-01-01

    Background: Nutritional supplements designed to increase adenosine 5'-triphosphate (ATP) concentrations are commonly used by athletes as ergogenic aids. ATP is the primary source of energy for the cells, and supplementation may enhance the ability to maintain high ATP turnover during high-intensity

  4. Adenosine 5 '-triphosphate (ATP) supplements are not orally bioavailable: a randomized, placebo-controlled cross-over trial in healthy humans

    NARCIS (Netherlands)

    Arts, I.C.W.; Coolen, E.J.C.M.; Bours, M.J.L.; Huyghebaert, N.; Cohen Stuart, M.A.; Bast, A.; Dagnelie, P.C.

    2012-01-01

    Background: Nutritional supplements designed to increase adenosine 5'-triphosphate (ATP) concentrations are commonly used by athletes as ergogenic aids. ATP is the primary source of energy for the cells, and supplementation may enhance the ability to maintain high ATP turnover during high-intensity

  5. Producing a highly concentrated coal suspension

    Energy Technology Data Exchange (ETDEWEB)

    Mokudzu, K.; Atsudzima, T.; Kiyedzuka, Y.

    1983-06-03

    Coal from wet and dry grinding is loaded into a mixer with a mixer arm with the acquisition of a highly concentrated suspension. Foamers (for instance, alkylbenzolsulfonate) and foam stabilizers (for instance diethanolamide of lauric acid) are added in a ratio of 10 to (2 to 5). The high fluidity of the suspension is maintained by injecting air into the suspension and an 80 percent concentration of the suspension is achieved.

  6. Fabricating high-energy quantum dots in ultra-thin LiFePO4 nanosheets using a multifunctional high-energy biomolecule-ATP

    DEFF Research Database (Denmark)

    Zhang, X.D.; Bi, Z.Y.; He, W.

    2014-01-01

    By using a multifunctional high-energy biomolecule—adenosine triphosphate (ATP)—we fabricated highenergy quantum dots (HEQDs) with a feature size of less than 10 nm and used them in high-power lithium-ion batteries. We introduced high-energy phosphate bonds into the crystal structure of LiFePO4...... coating network structures. This work is instructive for fabrication and design of new types of electrochemical energy conversion and storage devices with extraordinary properties and functions....

  7. Users' manual for the Langley high speed propeller noise prediction program (DFP-ATP)

    Science.gov (United States)

    Dunn, M. H.; Tarkenton, G. M.

    1989-01-01

    The use of the Dunn-Farassat-Padula Advanced Technology Propeller (DFP-ATP) noise prediction program which computes the periodic acoustic pressure signature and spectrum generated by propellers moving with supersonic helical tip speeds is described. The program has the capacity of predicting noise produced by a single-rotation propeller (SRP) or a counter-rotation propeller (CRP) system with steady or unsteady blade loading. The computational method is based on two theoretical formulations developed by Farassat. One formulation is appropriate for subsonic sources, and the other for transonic or supersonic sources. Detailed descriptions of user input, program output, and two test cases are presented, as well as brief discussions of the theoretical formulations and computational algorithms employed.

  8. Nanomolar ambient ATP decelerates P2X3 receptor kinetics.

    Science.gov (United States)

    Grote, Alexander; Hans, Michael; Boldogkoi, Zsolt; Zimmer, Andreas; Steinhäuser, Christian; Jabs, Ronald

    2008-12-01

    Homomeric P2X receptors differ in their electrophysiological and pharmacological profiles. The rapidly activating and desensitizing P2X3 receptors are known for their involvement in pain signalling pathways. Modulatory effects on P2X3 receptors have been reported for low concentrations of ATP ([ATP]). This includes both, enhancement and reduction of receptor currents. The first has been reported to be mediated by activation of ectoprotein kinases and high affinity desensitization (HAD), respectively. Both processes influence receptor current amplitudes. Here we describe a new phenomenon, the modulatory influence of ambient low [ATP] on P2X3 receptor kinetics. First, we studied in HEK cells whether persistent ATP affects current decay. To this end, P2X3 receptor mediated currents, elicited by pressure application of saturating [ATP], were analyzed after pre-application of low [ATP]. Second, UV-flash photolysis of ATP was employed to investigate whether submicromolar [ATP] affects receptor activation. Finally we confirmed the action of nanomolar [ATP] on native P2X3 receptors of neurons freshly isolated from rat dorsal root ganglia. We found that persistent low [ATP] caused pronounced deceleration of receptor current activation and decay. This priming effect indicates a mechanism different from HAD. It could be explained by a pre-opening receptor isomerization, induced by the occupation of a high affinity binding site already at the resting state. The observed modulation of the receptor kinetics could be considered as a physiological fine tuning mechanism of the nociceptive system, driven by the actual ambient agonist concentration.

  9. Study of an automatic ATP detecting system with high sensitivity based on bioluminescence%基于生物发光的高灵敏度ATP自动测试系统研究

    Institute of Scientific and Technical Information of China (English)

    岳伟伟; 何保山; 周爱玉; 罗金平; 蔡新霞

    2009-01-01

    To meet the needs of automatic adenosine triphosphate (ATP) detection with high sensitivity and wide response range, an ATP detection system was designed based on bioluminescence. By recording the light intensity at the beginning of the bioluminescence reaction using the automatic injection technology, the system can detect samples automatically with high sensitivity. The photo-electronic detection unit was optimized to decrease the influence of stray light and electromagnetism and increase the efficiency of luminescence detection. The circuit and data processing algorithm was optimized to broaden the detection range. The ATP samples with the concentration of 10-15~10-6 M were measured and the correlation coefficient between the ATP concentration and the light intensity was 0.974. The detection range was over 9 magnitudes. Compared with the commercial products, the system in this paper has the virtues of automatization, high sensitivity and wide detection range. Therefore, the ATP system has a great potential in ATP measurement and the corresponding fields such as bacteria, toxicity and hygienic conditions, and so on.%针对高灵敏度、宽响应范围及自动检测三磷酸腺苷(ATP)的需求,设计了一种基于生物发光原理的ATP检测系统.系统采用自动加样技术,在发光反应的同时开始记录光强,使仪表检测自动化,且提高检测灵敏度;在优化光电检测单元设计和减小杂散光及电磁干扰的同时,提高荧光检测效率;通过电路和数据处理算法优化提高仪表检测范围.以浓度为10-15~10-6 M范围内的标准ATP溶液进行了实验,结果表明仪表测量光强与ATP实际浓度相关系数为0.974,检测范围达9个数量级.与市场相关仪器相比,该系统具有自动化测试,性能和灵敏度高、响应范围宽的特点,在ATP测量以及相关领域如细菌测量、毒性测试及卫生状况检测等方面具有广泛的应用前景.

  10. Optogenetic control of ATP release

    Science.gov (United States)

    Lewis, Matthew A.; Joshi, Bipin; Gu, Ling; Feranchak, Andrew; Mohanty, Samarendra K.

    2013-03-01

    Controlled release of ATP can be used for understanding extracellular purinergic signaling. While coarse mechanical forces and hypotonic stimulation have been utilized in the past to initiate ATP release from cells, these methods are neither spatially accurate nor temporally precise. Further, these methods cannot be utilized in a highly effective cell-specific manner. To mitigate the uncertainties regarding cellular-specificity and spatio-temporal release of ATP, we herein demonstrate use of optogenetics for ATP release. ATP release in response to optogenetic stimulation was monitored by Luciferin-Luciferase assay (North American firefly, photinus pyralis) using luminometer as well as mesoscopic bioluminescence imaging. Our result demonstrates repetitive release of ATP subsequent to optogenetic stimulation. It is thus feasible that purinergic signaling can be directly detected via imaging if the stimulus can be confined to single cell or in a spatially-defined group of cells. This study opens up new avenue to interrogate the mechanisms of purinergic signaling.

  11. Extracellular ATP induces albuminuria in pregnant rats

    NARCIS (Netherlands)

    Faas, M.M.; van der Schaaf, G.; Borghuis, T.; Jongman, R.M.; van Pampus, Maria; de Vos, P.; van Goor, Harry; Bakker, W.W.

    2010-01-01

    BACKGROUND: As circulating plasma ATP concentrations are increased in pre-eclampsia, we tested whether increased plasma ATP is able to induce albuminuria during pregnancy. METHODS: Pregnant (day 14) and non-pregnant rats were infused with ATP (3000 microg/kg bw) via a permanent jugular vein cannula.

  12. Cellulase Inhibition by High Concentrations of Monosaccharides

    DEFF Research Database (Denmark)

    Hsieh, Chia-Wen; Cannella, David; Jørgensen, Henning

    2014-01-01

    that low free water availability contributes to cellulase inhibition. Of the hydrolytic enzymes involved, those acting on the cellulose substrate, that is, exo- and endoglucanases, were the most inhibited. The β -glucosidases were shown to be less sensitive to high monosaccharide concentrations except...

  13. High-dimensional entanglement concentration of twisted photon pairs High-dimensional entanglement concentration

    Science.gov (United States)

    Chen, L. X.; Wu, Q. P.

    2012-10-01

    Recently, Dada et al. reported on the experimental entanglement concentration and violation of generalized Bell inequalities with orbital angular momentum (OAM) [Nat. Phys. 7, 677 (2011)]. Here we demonstrate that the high-dimensional entanglement concentration can be performed in arbitrary OAM subspaces with selectivity. Instead of violating the generalized Bell inequalities, the working principle of present entanglement concentration is visualized by the biphoton OAM Klyshko picture, and its good performance is confirmed and quantified through the experimental Shannon dimensionalities after concentration.

  14. Design Optimization and Evaluation of a Bioluminescence Detection Part on a Microfluidic Device for in situ ATP Quantification

    Science.gov (United States)

    Aoki, Yusuke; Fukuba, Tatsuhiro; Yamamoto, Takatoki; Fujii, Teruo

    An integrated in situ analyzer for microbial ATP (IISA-ATP) has been developed with a microfluidic device as its core component to realize a compact and fully integrated system. In the system, a bioluminescence (luciferin—luciferase) reaction is conducted for ATP quantification. The microfluidic device has a coil-shaped microchannel for highly sensitive photo intensity measurement. In this paper, the concept of the IISA-ATP and optimization of the microchannel design to enhance sensitivity are presented. As a result of the optimization, linear correlation of the luminescence intensity with the ATP concentration in the range of 2 to 2 × 104 pM was achieved.

  15. Development of a high-throughput method to evaluate serum bactericidal activity using bacterial ATP measurement as survival readout

    Science.gov (United States)

    Saul, Allan; Rondini, Simona

    2017-01-01

    Serum Bactericidal Activity (SBA) assay is the method of choice to evaluate the complement-mediated functional activity of both infection- and vaccine-induced antibodies. To perform a typical SBA assay, serial dilutions of sera are incubated with target bacterial strains and complement. The conventional SBA assay is based on plating on agar the SBA reaction mix and counting the surviving bacterial colony forming units (CFU) at each serum dilution. Even with automated colony counting, it is labor-intensive, time-consuming and not amenable for large-scale studies. Here, we have developed a luminescence-based SBA (L-SBA) method able to detect surviving bacteria by measuring their ATP. At the end of the SBA reaction, a single commercially available reagent is added to each well of the SBA plate, and the resulting luminescence signal is measured in a microplate reader. The signal obtained is proportional to the ATP present, which is directly proportional to the number of viable bacteria. Bactericidal activity is subsequently calculated. We demonstrated the applicability of L-SBA with multiple bacterial serovars, from 5 species: Citrobacter freundii, Salmonella enterica serovars Typhimurium and Enteritidis, Shigella flexneri serovars 2a and 3a, Shigella sonnei and Neisseria meningitidis. Serum bactericidal titers obtained by the luminescence readout method strongly correlate with the data obtained by the conventional agar plate-based assay, and the new assay is highly reproducible. L-SBA considerably shortens assay time, facilitates data acquisition and analysis and reduces the operator dependency, avoiding the plating and counting of CFUs. Our results demonstrate that L-SBA is a useful high-throughput bactericidal assay. PMID:28192483

  16. ATP Release and Effects in Pancreas

    DEFF Research Database (Denmark)

    Novak, Ivana; Amstrup, Jan; Henriksen, Katrine Lütken

    2003-01-01

    ATP and other nucleotides are released from various cells, but the pathway and physiological stimulus for ATP release are often unclear. The focus of our studies is the understanding of ATP release and signaling in rat exocrine pancreas. In acinar suspension mechanical stimulation, hypotonic shock...... and, most importantly, cholinergic stimulation released 5-20nM ATP into the medium, as monitored by luminescence of the luciferin/luciferase reaction. ATP release was visualized at the single acinus level as luciferin consumption detected by confocal laser scanning microscopy (CLSM). The estimated ATP...... concentrations were higher, about 10µM, around acinar cells after cholinergic stimulation. Fluorescence of quinacrine and MANT-ATP indicated that some ATP is stored in secretory granules. Pancreatic acini have transcripts for P2X1, P2X4, P2Y2, and P2Y4 receptors, but measurements of Ca2+ signals in isolated...

  17. Estímulo no crescimento e na hidrólise de ATP em raízes de alface tratadas com humatos de vermicomposto: I - efeito da concentração Improving lettuce seedling root growth and ATP hydrolysis with humates from vermicompost: I - effect of vermicompost concentration

    Directory of Open Access Journals (Sweden)

    Maria Rita Cardoso Rodda

    2006-08-01

    Full Text Available O vermicomposto contém uma concentração elevada de substâncias húmicas e já é bem conhecido o efeito do seu uso sobre as propriedades do solo. No entanto, a ação direta das substâncias húmicas sobre o metabolismo das plantas é menos conhecida. O objetivo deste trabalho foi avaliar o uso de humatos extraídos de vermicomposto de esterco de curral com KOH 0,1 mol L-1 sobre o desenvolvimento e metabolismo de ATP em plântulas de alface. Após a germinação, plântulas de alface foram tratadas com os humatos em concentrações que variaram de 0 a 100 mg L-1 de C, durante quinze dias. Foram avaliados o crescimento da raiz e a atividade das bombas de H+ isoladas da fração microssomal do sistema radicular. Foi observado aumento na matéria fresca e seca do sistema radicular, bem como no número de sítios de mitose, raízes emergidas do eixo principal, na área e no comprimento radiculares, com o uso do humato na concentração de 25 mg L-1 de C. Também foi observado, nessa concentração, aumento significativo na hidrólise de ATP pelas bombas de H+, responsáveis pela geração de energia necessária à absorção de íons e pelo crescimento celular.Vermicomposts present a high content of humic substances and their effects on soil properties are well-documented. However, direct effects of humic substances on plant metabolism are less known. The aim of this work was to evaluate the use of humates isolated from cattle manure vermicompost with 0.1 mol L-1 KOH on root growth and ATP metabolism of lettuce seedlings. After germination, lettuce seedlings were treated with different humate concentrations (0 to 100 mg L-1 of C for 15 days. The root growth and the H+-ATPase activity of root cell microsomes isolated by differential centrifugation were evaluated. The humate treatment, at the concentration of 25 mg L-1 of C, increased fresh and dry root mass as well as root area and total length. Also there was a large increase in the number of

  18. Arabidopsis ATP A2 peroxidase. Expression and high-resolution structure of a plant peroxidase with implications for lignification

    DEFF Research Database (Denmark)

    Ostergaard, L; Teilum, K; Mirza, O;

    2000-01-01

    to be involved in lignin biosynthesis. Recently we isolated an extracellular anionic peroxidase, ATP A2, from rapidly lignifying Arabidopsis cell suspension culture and cloned its cDNA. Here we show that the Atp A2 promoter directs GUS reporter gene expression in lignified tissues of transgenic plants. Moreover......Lignins are phenolic biopolymers synthesized by terrestrial, vascular plants for mechanical support and in response to pathogen attack. Peroxidases have been proposed to catalyse the dehydrogenative polymerization of monolignols into lignins, although no specific isoenzyme has been shown......-coumaryl and coniferyl alcohols are preferred by ATP A2, while the oxidation of sinapyl alcohol will be sterically hindered in ATP A2 as well as in all other plant peroxidases due to an overlap with the conserved Pro-139. We suggest ATP A2 is involved in a complex regulation of the covalent cross-linking in the plant...

  19. High glucose decreases the expression of ATP-binding cassette transporter G1 in human vascular smooth muscle cells

    Institute of Scientific and Technical Information of China (English)

    Jiahong Xue; Zuyi Yuan; Yue Wu; Yan Zhao; Zhaofei Wan

    2008-01-01

    Objective:ATP-binding cassette transporters(ABC) A1 and G1 play an important role in mediating cholesterol efflux and preventing macrophage foam cell formation. In this study, we examined the regulation of ABC transporters by high glucose in human vascular smooth muscle cells(VSMCs), the other precursor of foam cells. Methods:Incubation of human VSMCs with D-ghicose(5 to 30 mM) for 1 to 7 days in the presence or absence of antioxidant and nuclear factor(NF)-kB inhibitors, the expressions of ABCA1 and ABCG1 were analyzed by real time PCR and Western blotting. Results:High glucose decreased ABCG1 mRNA and protein expression in cultured VSMCs, whereas the expression of ABCA1 was not significantly decreased. Down-regulation of ABCG1 mRNA expression by high glucose was abolished by antioxidant N-acetyl-L-cysteine(NAC) and NF-kB inhibitors, BAY 11-7085 and tosyl-phenylalanine chloromethyl-ketone(TPCK). Conclusion:High glucose suppresses the expression of ABCG1 in VSMCs, which is the possible mechanism of VSMC derived foam cell transformation.

  20. Dual recognition unit strategy improves the specificity of the adenosine triphosphate (ATP) aptamer biosensor for cerebral ATP assay.

    Science.gov (United States)

    Yu, Ping; He, Xiulan; Zhang, Li; Mao, Lanqun

    2015-01-20

    Adenosine triphosphate (ATP) aptamer has been widely used as a recognition unit for biosensor development; however, its relatively poor specificity toward ATP against adenosine-5'-diphosphate (ADP) and adenosine-5'-monophosphate (AMP) essentially limits the application of the biosensors in real systems, especially in the complex cerebral system. In this study, for the first time, we demonstrate a dual recognition unit strategy (DRUS) to construct a highly selective and sensitive ATP biosensor by combining the recognition ability of aptamer toward A nucleobase and of polyimidazolium toward phosphate. The biosensors are constructed by first confining the polyimidazolium onto a gold surface by surface-initiated atom transfer radical polymerization (SI-ATRP), and then the aptamer onto electrode surface by electrostatic self-assembly to form dual-recognition-unit-functionalized electrodes. The constructed biosensor based on DRUS not only shows an ultrahigh sensitivity toward ATP with a detection limit down to the subattomole level but also an ultrahigh selectivity toward ATP without interference from ADP and AMP. The constructed biosensor is used for selective and sensitive sensing of the extracellular ATP in the cerebral system by combining in vivo microdialysis and can be used as a promising neurotechnology to probing cerebral ATP concentration.

  1. Arabidopsis ATP A2 peroxidase. Expression and high-resolution structure of a plant peroxidase with implications for lignification

    DEFF Research Database (Denmark)

    Ostergaard, L; Teilum, K; Mirza, O

    2000-01-01

    to be involved in lignin biosynthesis. Recently we isolated an extracellular anionic peroxidase, ATP A2, from rapidly lignifying Arabidopsis cell suspension culture and cloned its cDNA. Here we show that the Atp A2 promoter directs GUS reporter gene expression in lignified tissues of transgenic plants. Moreover......, an Arabidopsis mutant with increased lignin levels compared to wild type shows increased levels of ATP A2 mRNA and of a mRNA encoding an enzyme upstream in the lignin biosynthetic pathway. The substrate specificity of ATP A2 was analysed by X-ray crystallography and docking of lignin precursors. The structure...... of ATP A2 was solved to 1.45 A resolution at 100 K. Docking of p-coumaryl, coniferyl and sinapyl alcohol in the substrate binding site of ATP A2 were analysed on the basis of the crystal structure of a horseradish peroxidase C-CN-ferulic acid complex. The analysis indicates that the precursors p...

  2. Diverse microbial species survive high ammonia concentrations

    Science.gov (United States)

    Kelly, Laura C.; Cockell, Charles S.; Summers, Stephen

    2012-04-01

    Planetary protection regulations are in place to control the contamination of planets and moons with terrestrial micro-organisms in order to avoid jeopardizing future scientific investigations relating to the search for life. One environmental chemical factor of relevance in extraterrestrial environments, specifically in the moons of the outer solar system, is ammonia (NH3). Ammonia is known to be highly toxic to micro-organisms and may disrupt proton motive force, interfere with cellular redox reactions or cause an increase of cell pH. To test the survival potential of terrestrial micro-organisms exposed to such cold, ammonia-rich environments, and to judge whether current planetary protection regulations are sufficient, soil samples were exposed to concentrations of NH3 from 5 to 35% (v/v) at -80°C and room temperature for periods up to 11 months. Following exposure to 35% NH3, diverse spore-forming taxa survived, including representatives of the Firmicutes (Bacillus, Sporosarcina, Viridibacillus, Paenibacillus, Staphylococcus and Brevibacillus) and Actinobacteria (Streptomyces). Non-spore forming organisms also survived, including Proteobacteria (Pseudomonas) and Actinobacteria (Arthrobacter) that are known to have environmentally resistant resting states. Clostridium spp. were isolated from the exposed soil under anaerobic culture. High NH3 was shown to cause a reduction in viability of spores over time, but spore morphology was not visibly altered. In addition to its implications for planetary protection, these data show that a large number of bacteria, potentially including spore-forming pathogens, but also environmentally resistant non-spore-formers, can survive high ammonia concentrations.

  3. Detecting ATP release by a biosensor method.

    Science.gov (United States)

    Hayashi, Seiji; Hazama, Akihiro; Dutta, Amal K; Sabirov, Ravshan Z; Okada, Yasunobu

    2004-11-09

    Cells release adenosine 5'-triphosphate (ATP) into the extracellular space in response to various stimuli. This released ATP plays an important physiological role in cell-to-cell signal transduction. The bulk ATP concentration can be detected using a conventional luciferin-luciferase assay. However, the ATP concentration in the vicinity of the cell surface is often different from the bulk concentration because of its rapid degradation by ecto-ATPases and because of delayed diffusion due to unstirred layer effects. Here, we describe a simple biosensor method to measure the local ATP concentration on the cell surface in real time. The method is based on the ATP-dependent opening of ligand-gated cation channels of purinergic P2X receptors expressed in undifferentiated pheochromocytoma (PC12) cells or in human embryonic kidney 293 (HEK293) cells stably transfected with recombinant P2X2 purinergic receptors. Under the whole-cell configuration of patch-clamp, a sensor PC12 cell or HEK293 is positioned within the proximity of a target cell, and the P2X-mediated currents induced by ATP released from a given site on the target cell surface is measured. The ATP release is quantified by a calibration procedure utilizing local puff applications of ATP at preset concentrations.

  4. Effect of dietary selenium and vitamin E on the ultrastructure and ATP concentration of boar spermatozoa, and the efficacy of added sodium selenite in extended semen on sperm motility.

    Science.gov (United States)

    Marin-Guzman, J; Mahan, D C; Whitmoyer, R

    2000-06-01

    Three experiments evaluated the effects of dietary Se and vitamin E on the ultrastructure of spermatozoa, ATP concentration of spermatozoa, and the effects of adding sodium selenite to semen extenders on subsequent sperm motility. The experiment was a 2 x 2 arrangement of treatments in a randomized complete block design. A total of 10 mature boars were fed from weaning to 18 mo of age diets fortified with two levels of supplemental Se (0 or .5 ppm) or vitamin E (0 or 220 IU/kg diet). The nonfortified diets contained .06 ppm Se and 4.4 IU vitamin E/kg. In Exp. 1, the spermatozoa from all boars were examined by electron microscopy. Vitamin E had no effect on structural abnormalities in the spermatozoa. When the low-Se diet was fed the acrosome or nuclei of the spermatozoa was unaffected, but the mitochondria in the tail midpiece were more oval with wider gaps between organelles. The plasma membrane connection to the tail midpiece was not tightly bound as when boars were fed Se. Immature spermatozoa with cytoplasmic droplets were more numerous when boars were fed the low-Se diet, but the occurrence of midpiece abnormalities occurred in boars fed diets with or without Se or vitamin E. Our results suggest that Se may enhance spermatozoa maturation in the epididymis and may reduce the number of sperm with cytoplasmic droplets. In Exp. 2, the concentration of ATP in the spermatozoa was evaluated in the semen of all treatment boars. When the low-Se diet was fed, ATP concentration was lower (P spermatozoa. Overall, these results suggest that low Se-diets fed to boars resulted in abnormal spermatozoal mitochondria, a lower ATP concentration in the spermatozoa, and a loose apposition of the plasma membrane to the helical coil of the tail midpiece, but no effect from inadequate vitamin E was demonstrated. Adding sodium selenite to the semen extender reduced sperm cell motility.

  5. Performance analysis of surge arrester on high voltage systems using ATP

    Energy Technology Data Exchange (ETDEWEB)

    Nallagownden, P.; Magumane, A.H. [Univ. Teknologi Petronas, Perak (Malaysia); Kanth, K.S.R. [Tenaga National Berhad (Malaysia)

    2008-07-01

    Lightning strikes are among the major factors that cause failures in electrical power systems. Phase to ground arresters are commonly installed at power transformer terminals to offer some lightning protection. However, it is important to understand the performance of metal oxide arresters under very fast transient overvoltages in order to determine the protection zone of the arrester and to achieve economical benefits. This study investigated lightning overvoltage protection in a complete three-phase scheme of a 500 KV substation. Overvoltages originated from direct lightning stroke on a phase of a real overhead line (OHL) model. The effect of the separation distance of the arrester from the transformer connected at the open end of the substations was investigated as well as the performance of the arrester for different substation configurations. In the first scenario, the connection of the arrester and transformer was done with a direct connection of an overhead line. In the second scenario, the connection of these devices was done through a cable. Both the overhead line and cable lengths were varied and the maximum overvoltages coming to the transformer were recorded. The results showed that there is a direct proportionality between overvoltages and length of the overhead line or cable. As long the length of the line or cable between the arrester and the transformer was increased, the vulnerability of the transformer to receive high overvoltages also increased. Surge overvoltages were found to be very sensitive to impedance of the line or cable. The direct connections of overhead lines between the arrester and transformer make it necessary to add some protective device. It was suggested that surge arresters should be installed every 200 meters along the overhead lines in order to ensure the safety of equipment. 12 refs., 4 tabs., 8 figs.

  6. Online cleanup of accelerated solvent extractions for determination of adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), and adenosine 5'-monophosphate (AMP) in royal jelly using high-performance liquid chromatography.

    Science.gov (United States)

    Xue, Xiaofeng; Wang, Feng; Zhou, Jinhui; Chen, Fang; Li, Yi; Zhao, Jing

    2009-06-10

    Determination of the levels of adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), and adenosine 5'-monophosphate (AMP) in royal jelly is important for the study of its pharmacological activities, health benefits, and adenosine phosphate degradation. In this study was developed a novel method to determine ATP, ADP, and AMP levels in royal jelly using accelerated solvent extraction (ASE) followed by online cleanup and high-performance liquid chromatography (HPLC) with diode array detection (DAD). The optimum extraction conditions were obtained using an 11 mL ASE cell, ethanol/water (5:5 v/v) as the extraction solvent, 1500 psi, 80 degrees C, a 5 min static time, and a 60% flush volume. Optimum separation of the three compounds was achieved in AMP levels in 15 samples of royal jelly of different origins was performed. Sample results indicated that the AMP concentration was 24.2-2214.4 mg kg(-1), whereas ATP and ADP were not detectable or present only at low levels.

  7. Physical chemistry of highly concentrated emulsions.

    Science.gov (United States)

    Foudazi, Reza; Qavi, Sahar; Masalova, Irina; Malkin, Alexander Ya

    2015-06-01

    This review explores the physics underlying the rheology of highly concentrated emulsions (HCEs) to determine the relationship between elasticity and HCE stability, and to consider whether it is possible to describe all physicochemical properties of HCEs on the basis of a unique physical approach. We define HCEs as emulsions with a volume fraction above the maximum closest packing fraction of monodisperse spheres, φm=0.74, even if droplets are not of polyhedron shape. The solid-like rheological behavior of HCEs is characterized by yield stress and elasticity, properties which depend on droplet polydispersity and which are affected by caging at volume fractions about the jamming concentration, φj. A bimodal size distribution in HCEs diminishes caging and facilitates droplet movement, resulting in HCEs with negligible yield stress and no plateau in storage modulus. Thermodynamic forces automatically move HCEs toward the lowest free energy state, but since interdroplet forces create local minimums - points beyond which free energy temporarily increases before it reaches the global minimum of the system - the free energy of HCEs will settle at a local minimum unless additional energy is added. Several attempts have been undertaken to predict the elasticity of HCEs. In many cases, the elastic modulus of HCEs is higher than the one predicted from classical models, which only take into account spatial repulsion (or simply interfacial energy). Improved models based on free energy calculation should be developed to consider the disjoining pressure and interfacial rheology in addition to spatial repulsion. The disjoining pressure and interfacial viscoelasticity, which result in the deviation of elasticity from the classical model, can be regarded as parameters for quantifying the stability of HCEs. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Modification of the ATP inhibitory site of the Ascaris suum phosphofructokinase results in the stabilization of an inactive T state

    Energy Technology Data Exchange (ETDEWEB)

    Rao, G.S.J.; Cook, P.F.; Harris, B.G. (Univ. of North Texas, Fort Worth (United States))

    1991-10-15

    Treatment of the Ascaris suum phosphofructokinase (PFK) with 2{prime},3{prime}-dialdehyde ATP (oATP) results in an enzyme form that is inactive. The conformational integrity of the active site, however, is preserved, suggesting that oATP modification locks the PFK into an inactive T state that cannot be activated. A rapid, irreversible first-order inactivation of the PFK is observed in the presence of oATP. The rate of inactivation is saturable and gives a K{sub oATP} of 1.07 {plus minus} 0.27 mM. Complete protection against inactivation is afforded by high concentrations of ATP. This desensitized enzyme incorporates only 0.2-0.3 mol of ({sup 3}H)oATP/subunit, suggesting that in te native enzyme inactivation perhaps results from the modification of the ATP inhibitory site rather than the catalytic site. Modification of an active-site thiol by 4,4{prime}-dithiodipyridine is prevented yb ATP before and after oATP treatment. Finally, gel filtration HPLC studies show that the oATP-modified enzyme retains its tetrameric state and neither the tryptophan fluorescence nor the circular dichroic spectra of the modified enzyme are affected by fructose 2,6-bisphosphate, suggesting that the enzyme is locked into a tetrameric inactive T state.

  9. ATP Production in Chlamydomonas reinhardtii Flagella by Glycolytic Enzymes

    DEFF Research Database (Denmark)

    Mitchell, Beth F; Pedersen, Lotte B; Feely, Michael

    2005-01-01

    Eukaryotic cilia and flagella are long, thin organelles, and diffusion from the cytoplasm may not be able to support the high ATP concentrations needed for dynein motor activity. We discovered enzyme activities in the Chlamydomonas reinhardtii flagellum that catalyze three steps of the lower half...... of glycolysis (phosphoglycerate mutase, enolase, and pyruvate kinase). These enzymes can generate one ATP molecule for every substrate molecule consumed. Flagellar fractionation shows that enolase is at least partially associated with the axoneme, whereas phosphoglycerate mutase and pyruvate kinase primarily...... reside in the detergent-soluble (membrane + matrix) compartments. We further show that axonemal enolase is a subunit of the CPC1 central pair complex and that reduced flagellar enolase levels in the cpc1 mutant correlate with the reduced flagellar ATP concentrations and reduced in vivo beat frequencies...

  10. High-Temperature, High-Concentration Solar Thermoelectric Generators

    Science.gov (United States)

    Warren, Emily; Baranowski, Lauryn; Olsen, Michele; Ndione, Paul; Netter, Judy; Goodrich, Alan; Gray, Matthew; Parilla, Philip; Ginley, David; Toberer, Eric

    2014-03-01

    Solar thermoelectric generators (STEGs) powered with concentrated solar energy have potential for use as primary energy converters or as topping-cycles for more conventional concentrated solar power (CSP) technologies. Modeling based on current record modules from JPL suggests thermoelectric efficiencies of 18 % could be experimentally expected with a temperature gradient of 1000 - 100°C. Integrating these state-of-the-art TEGs with a concentrating solar receiver requires simultaneous optimization of optical, thermal, and thermoelectric systems. This talk will discuss the modeling, design, and experimental testing of STEG devices under concentrated sunlight. We have developed a model that combines thermal circuit modeling with optical ray tracing to design selective absorber coatings and cavities to minimize radiation losses from the system. We have fabricated selective absorber coatings and demonstrated that these selective absorber films can minimize blackbody radiation losses at high temperature and are stable after thermal cycling to 1000°C. On-sun testing of STEG devices and thermal simulators is ongoing and preliminary results will be discussed.

  11. Reactions of a fluorescent ATP analog, 2'-(5-dimethyl-aminonaphthalene-1-sulfonyl) amino-2'-deoxyATP, with E. coli F1-ATPase and its subunits: the roles of the high affinity binding site in the alpha subunit and the low affinity binding site in the beta subunit.

    Science.gov (United States)

    Matsuoka, I; Takeda, K; Futai, M; Tonomura, Y

    1982-11-01

    . The kinetic properties of the fluorescence change of DNS-ATP in the reaction with the reconstituted EF1-ATPase were quite similar to those of native EF1. Most of our findings are consistent with a simple mechanism that the high affinity catalytic site and low affinity regulatory site exist in the alpha subunit and beta subunit, respectively. However, the findings mentioned in (4) suggest that the binding of the alpha and beta subunit, which is mediated by the gamma subunit, induces conformational change(s) in the ATP binding site located probably in the alpha subunit, and that the conformational change(s) is essential to exert the full hydrolyzing activity.

  12. Production of high concentrations of yeast

    Energy Technology Data Exchange (ETDEWEB)

    1981-11-10

    A microbe is aerobically cultured using O/sub 2/ or a gas rich in O/sub 2/. The grown cells are washed, concentrated and a portion of the cells used as a seed culture. Thus, Saccharomyces cerevisiae (bakers' yeast) was cultured in a jar fermentor by flow down system maintaining the dissolved O/sub 2/ at 2-5 mg/L; volume of the initial medium containing 30% glucose was 350 mL and the initial washed cell concentration was 50 g dry cells/L. After 12 hours of cultivation, the volume of the medium increased to 750 mL and the cell concentration rose to 102 g dry cells/L; the yield was 49% with respect to glucose. The cells were washed and the cultivation was repeated by use of the washed cells; cell concentration reached 105 g dry cells/L.

  13. Horizontally staggered lightguide solar concentrator with lateral displacement tracking for high concentration applications.

    Science.gov (United States)

    Ma, Hongcai; Wu, Lin

    2015-07-10

    We present the design of a horizontally staggered lightguide solar concentrator with lateral displacement tracking for high concentration applications. This solar concentrator consists of an array of telecentric primary concentrators, a horizontally staggered lightguide layer, and a vertically tapered lightguide layer. The primary concentrator is realized by two plano-aspheric lenses with lateral movement and maintains a high F-number over an angle range of ±23.5°. The results of the simulations show that the solar concentrator achieves a high concentration ratio of 500× with ±0.5° of acceptance angle by a single-axis tracker and dual lateral translation stages.

  14. Macula densa basolateral ATP release is regulated by luminal [NaCl] and dietary salt intake.

    Science.gov (United States)

    Komlosi, Peter; Peti-Peterdi, Janos; Fuson, Amanda L; Fintha, Attila; Rosivall, Laszlo; Bell, Phillip Darwin

    2004-06-01

    One component of the macula densa (MD) tubuloglomerular feedback (TGF) signaling pathway may involve basolateral release of ATP through a maxi-anion channel. Release of ATP has previously been studied during a maximal luminal NaCl concentration ([NaCl](L)) stimulus (20-150 mmol/l). Whether MD ATP release occurs during changes in [NaCl](L) within the physiological range (20-60 mmol/l) has not been examined. Also, because TGF is known to be enhanced by low dietary salt intake, we examined the pattern of MD ATP release from salt-restricted rabbits. Fluorescence microscopy, with fura 2-loaded cultured mouse mesangial cells as biosensors, was used to assess ATP release from the isolated, perfused thick ascending limb containing the MD segment. The mesangial biosensor cells, which contain purinergic receptors and elevate intracellular Ca(2+) concentration ([Ca(2+)](i)) on ATP binding, were placed adjacent to the MD basolateral membrane. Elevations in [NaCl](L) between 0 and 80 mmol/l, in 20-mmol/l increments, caused stepwise increases in [Ca(2+)](i), with the highest increase at [NaCl](L) of approximately 60 mmol/l. Luminal furosemide at 10(-4) mol/l blocked ATP release, which suggests that the efflux of ATP required MD Na-2Cl-K cotransport. A low-salt diet for 1 wk increased the magnitude of [NaCl](L)-dependent elevations in biosensor [Ca(2+)](i) by twofold, whereas high-salt intake had no effect. In summary, ATP release occurs over the same range of [NaCl](L) (20-60 mmol/l) previously reported for TGF responses, and, similar to TGF, ATP release was enhanced by dietary salt restriction. Thus these two findings are consistent with the role of MD ATP release as a signaling component of the TGF pathway.

  15. HMR 1098 is not an SUR isotype specific inhibitor of heterologous or sarcolemmal K ATP channels.

    Science.gov (United States)

    Zhang, Hai Xia; Akrouh, Alejandro; Kurata, Harley T; Remedi, Maria Sara; Lawton, Jennifer S; Nichols, Colin G

    2011-03-01

    Murine ventricular and atrial ATP-sensitive potassium (K(ATP)) channels contain different sulfonylurea receptors (ventricular K(ATP) channels are Kir6.2/SUR2A complexes, while atrial K(ATP) channels are Kir6.2/SUR1 complexes). HMR 1098, the sodium salt of HMR 1883 {1-[[5-[2-(5-chloro-o-anisamido)ethyl]-2-methoxyphenyl]sulfonyl]-3-methylthiourea}, has been considered as a selective sarcolemmal (i.e. SUR2A-dependent) K(ATP) channel inhibitor. However, it is not clear whether HMR 1098 would preferentially inhibit ventricular K(ATP) channels over atrial K(ATP) channels. To test this, we used whole-cell patch clamp techniques on mouse atrial and ventricular myocytes as well as (86)Rb(+) efflux assays and excised inside-out patch clamp techniques on Kir6.2/SUR1 and Kir6.2/SUR2A channels heterologously expressed in COSm6 cells. In mouse atrial myocytes, both spontaneously activated and diazoxide-activated K(ATP) currents were effectively inhibited by 10 μM HMR 1098. By contrast, in ventricular myocytes, pinacidil-activated K(ATP) currents were inhibited by HMR 1098 at a high concentration (100 μM) but not at a low concentration (10 μM). Consistent with this finding, HMR 1098 inhibits (86)Rb(+) effluxes through Kir6.2/SUR1 more effectively than Kir6.2/SUR2A channels in COSm6 cells. In excised inside-out patches, HMR 1098 inhibited Kir6.2/SUR1 channels more effectively, particularly in the presence of MgADP and MgATP (mimicking physiological stimulation). Finally, dose-dependent enhancement of insulin secretion from pancreatic islets and decrease of blood glucose level confirm that HMR 1098 is an inhibitor of Kir6.2/SUR1-composed K(ATP) channels.

  16. ATP-Competitive Inhibitors of the Mammalian Target of Rapamycin: Design and Synthesis of Highly Potent and Selective Pyrazolopyrimidines

    Energy Technology Data Exchange (ETDEWEB)

    Zask, Arie; Verheijen, Jeroen C.; Curran, Kevin; Kaplan, Joshua; Richard, David J.; Nowak, Pawel; Malwitz, David J.; Brooijmans, Natasja; Bard, Joel; Svenson, Kristine; Lucas, Judy; Toral-Barza, Lourdes; Zhang, Wei-Guo; Hollander, Irwin; Gibbons, James J.; Abraham, Robert T.; Ayral-Kaloustian, Semiramis; Mansour, Tarek S.; Yu, Ker; (Wyeth)

    2009-09-18

    The mammalian target of rapamycin (mTOR), a central regulator of growth, survival, and metabolism, is a validated target for cancer therapy. Rapamycin and its analogues, allosteric inhibitors of mTOR, only partially inhibit one mTOR protein complex. ATP-competitive, global inhibitors of mTOR that have the potential for enhanced anticancer efficacy are described. Structural features leading to potency and selectivity were identified and refined leading to compounds with in vivo efficacy in tumor xenograft models.

  17. Urinary ATP may be a dynamic biomarker of detrusor overactivity in women with overactive bladder syndrome.

    Directory of Open Access Journals (Sweden)

    Miguel Silva-Ramos

    Full Text Available BACKGROUND: Nowadays, there is a considerable bulk of evidence showing that ATP has a prominent role in the regulation of human urinary bladder function and in the pathophysiology of detrusor overactivity. ATP mediates nonadrenergic-noncholinergic detrusor contractions in overactive bladders. In vitro studies have demonstrated that uroepithelial cells and cholinergic nerves from overactive human bladder samples (OAB release more ATP than controls. Here, we compared the urinary ATP concentration in samples collected non-invasively from OAB women with detrusor overactivity and age-matched controls. METHODS: Patients with neurologic diseases, history of malignancy, urinary tract infections or renal impairment (creatinine clearance <70 ml/min were excluded. All patients completed a 3-day voiding diary, a 24 h urine collection and blood sampling to evaluate creatinine clearance. Urine samples collected during voluntary voids were immediately freeze-preserved for ATP determination by the luciferin-luciferase bioluminescence assay; for comparison purposes, samples were also tested for urinary nerve growth factor (NGF by ELISA. RESULTS: The urinary content of ATP, but not of NGF, normalized to patients' urine creatinine levels (ATP/Cr or urinary volume (ATP.Vol were significantly (P<0.05 higher in OAB women with detrusor overactivity (n = 34 than in healthy controls (n = 30. Significant differences between the two groups were still observed by boosting urinary ATP/Cr content after water intake, but these were not detected for NGF/Cr. In OAB patients, urinary ATP/Cr levels correlated inversely with mean voided volumes determined in a 3-day voiding diary. CONCLUSION: A high area under the receiver operator characteristics (ROC curve (0.741; 95% CI 0.62-0.86; P<0.001 is consistent with urinary ATP/Cr being a highly sensitive dynamic biomarker for assessing detrusor overactivity in women with OAB syndrome.

  18. Adenosine 5′-triphosphate (ATP supplements are not orally bioavailable: a randomized, placebo-controlled cross-over trial in healthy humans

    Directory of Open Access Journals (Sweden)

    Arts Ilja CW

    2012-04-01

    Full Text Available Abstract Background Nutritional supplements designed to increase adenosine 5′-triphosphate (ATP concentrations are commonly used by athletes as ergogenic aids. ATP is the primary source of energy for the cells, and supplementation may enhance the ability to maintain high ATP turnover during high-intensity exercise. Oral ATP supplements have beneficial effects in some but not all studies examining physical performance. One of the remaining questions is whether orally administered ATP is bioavailable. We investigated whether acute supplementation with oral ATP administered as enteric-coated pellets led to increased concentrations of ATP or its metabolites in the circulation. Methods Eight healthy volunteers participated in a cross-over study. Participants were given in random order single doses of 5000 mg ATP or placebo. To prevent degradation of ATP in the acidic environment of the stomach, the supplement was administered via two types of pH-sensitive, enteric-coated pellets (targeted at release in the proximal or distal small intestine, or via a naso-duodenal tube. Blood ATP and metabolite concentrations were monitored by HPLC for 4.5 h (naso-duodenal tube or 7 h (pellets post-administration. Areas under the concentration vs. time curve were calculated and compared by paired-samples t-tests. Results ATP concentrations in blood did not increase after ATP supplementation via enteric-coated pellets or naso-duodenal tube. In contrast, concentrations of the final catabolic product of ATP, uric acid, were significantly increased compared to placebo by ~50% after administration via proximal-release pellets (P = 0.003 and naso-duodenal tube (P = 0.001, but not after administration via distal-release pellets. Conclusions A single dose of orally administered ATP is not bioavailable, and this may explain why several studies did not find ergogenic effects of oral ATP supplementation. On the other hand, increases in uric acid after release of

  19. OBSERVATIONS ON AN ATP-SENSITIVE PROTEIN SYSTEM FROM THE PLASMODIA OF A MYXOMYCETE

    Science.gov (United States)

    Ts'o, Paul O. P.; Bonner, James; Eggman, Luther; Vinograd, Jerome

    1956-01-01

    1. Extracts of the plasmodia of the myxomycete, Physarum polycephalum, exhibit reversible decreases in viscosity in response to the addition of ATP under appropriate conditions. The protoplasm material prepared by extraction with KCl solution can apparently exist in either a high or a low viscosity state. As prepared, it is in the low viscosity condition. Rapid and extensive increases in viscosity of the extract are brought about by addition of AMP, inorganic phosphate, or, under certain conditions, of ATP. Only after the high viscosity state has been attained does addition of appropriate quantities of ATP cause a reversible decrease in viscosity. 2. The active principle of crude plasmodial extracts may be concentrated by fractional precipitation with ammonium sulfate and is found in the fraction precipitated between 30 and 40 per cent saturation. This material possesses a higher viscosity than does the original crude extract and is apparently in the high viscosity state since the addition of ATP causes an immediate reversible decrease in viscosity. 3. The ATP-sensitive fraction of myxomycete plasmodia possesses a viscosity which is dependent upon its previous thermal treatment. Extracts incubated at 0° for a period of a few hours increase greatly in viscosity when they are returned to 24.5°. This increased viscosity is structural in nature, is destroyed by mechanical agitation of the solution, and may be reversibly destroyed by addition of ATP. 4. It is suggested that the ATP-responsive protein of myxomycete plasmodia may be related to sol-gel transformations which have been observed in intact plasmodia and may participate in the protoplasmic streaming of the intact organism. This suggestion is based upon the following facts: (a) the protoplasmic streaming of myxomycete plasmodia is increased by microinjection of ATP; (b) the gel portion of the cytoplasm at the site of the microinjection of ATP is extensively converted to the sol state. The changes in structure

  20. Imaging Adenosine Triphosphate (ATP).

    Science.gov (United States)

    Rajendran, Megha; Dane, Eric; Conley, Jason; Tantama, Mathew

    2016-08-01

    Adenosine triphosphate (ATP) is a universal mediator of metabolism and signaling across unicellular and multicellular species. There is a fundamental interdependence between the dynamics of ATP and the physiology that occurs inside and outside the cell. Characterizing and understanding ATP dynamics provide valuable mechanistic insight into processes that range from neurotransmission to the chemotaxis of immune cells. Therefore, we require the methodology to interrogate both temporal and spatial components of ATP dynamics from the subcellular to the organismal levels in live specimens. Over the last several decades, a number of molecular probes that are specific to ATP have been developed. These probes have been combined with imaging approaches, particularly optical microscopy, to enable qualitative and quantitative detection of this critical molecule. In this review, we survey current examples of technologies available for visualizing ATP in living cells, and identify areas where new tools and approaches are needed to expand our capabilities.

  1. High concentrations of H2O2 make aerobic glycolysis energetically more favourable than cellular respiration.

    Directory of Open Access Journals (Sweden)

    Hamid R Molavian

    2016-08-01

    Full Text Available Since the original observation of the Warburg Effect in cancer cells, over eight decades ago, the major question of why aerobic glycolysis is favored over oxidative phosphorylation has remained unresolved. An understanding of this phenomenon may well be the key to the development of more effective cancer therapies. In this paper, we use a semi-empirical method to throw light on this puzzle. We show that aerobic glycolysis is in fact energetically more favorable than oxidative phosphorylation for concentrations of peroxide (H2O2 above some critical threshold value. The fundamental reason for this is the activation and high engagement of the pentose phosphate pathway (PPP in response to the production of reactive oxygen species H2O2 by mitochondria and the high concentration of H2O2 (produced by mitochondria and other sources. This makes oxidative phosphorylation an inefficient source of energy since it leads (despite high levels of ATP production to a concomitant high energy consumption in order to respond to the hazardous waste products resulting from cellular processes associated with this metabolic pathway. We also demonstrate that the high concentration of H2O2 results in an increased glucose consumption, and also increases the lactate production in the case of glycolysis.

  2. Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

    Directory of Open Access Journals (Sweden)

    Quynh Thu Nguyen

    2016-11-01

    Full Text Available Mammalian spermatozoa utilize ATP as the energy source for key functions on the route to fertilization. ATP and its precursor nucleotides ADP and AMP are regularly investigated in sperm physiology studies, mostly by bioluminescence assays. Assay results vary widely, mainly due to different efficiencies in nucleotide extraction and prevention of their enzymatic degradation. Here, we describe a revised, validated protocol for efficient phosphatase inhibition and adenine nucleotide extraction resulting in consistently high ATP concentrations exceeding previously reported values for boar spermatozoa up to 20-fold. The revised assay is applicable for determining ATP concentrations and adenylate energy charge in extracts from fresh and frozen samples, thereby allowing simultaneous assessment of semen samples from long-term storage experiments. After validation, the assay was applied to liquid-preserved boar spermatozoa stored at 17°C and 5°C for 24 and 72 h. Cooling to 5°C, but not storage duration, reduced ATP concentration in spermatozoa (P<0.05, which was accompanied by the appearance of AMP and ADP in the preservation medium. ATP and energy charge were highly correlated to the proportion of membrane-intact spermatozoa, supporting the idea of nucleotides leaking through disrupted membranes in cold-shocked cells. The present assay allows highly standardized studies of energy metabolism in spermatozoa.

  3. Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

    Science.gov (United States)

    Nguyen, Quynh Thu; Wallner, Ulrike; Schmicke, Marion; Waberski, Dagmar

    2016-01-01

    ABSTRACT Mammalian spermatozoa utilize ATP as the energy source for key functions on the route to fertilization. ATP and its precursor nucleotides ADP and AMP are regularly investigated in sperm physiology studies, mostly by bioluminescence assays. Assay results vary widely, mainly due to different efficiencies in nucleotide extraction and prevention of their enzymatic degradation. Here, we describe a revised, validated protocol for efficient phosphatase inhibition and adenine nucleotide extraction resulting in consistently high ATP concentrations exceeding previously reported values for boar spermatozoa up to 20-fold. The revised assay is applicable for determining ATP concentrations and adenylate energy charge in extracts from fresh and frozen samples, thereby allowing simultaneous assessment of semen samples from long-term storage experiments. After validation, the assay was applied to liquid-preserved boar spermatozoa stored at 17°C and 5°C for 24 and 72 h. Cooling to 5°C, but not storage duration, reduced ATP concentration in spermatozoa (P<0.05), which was accompanied by the appearance of AMP and ADP in the preservation medium. ATP and energy charge were highly correlated to the proportion of membrane-intact spermatozoa, supporting the idea of nucleotides leaking through disrupted membranes in cold-shocked cells. The present assay allows highly standardized studies of energy metabolism in spermatozoa. PMID:27612509

  4. Effect of Mg(2+)-ATP on acetylcholinesterase of Electrophorus electricus (L.).

    Science.gov (United States)

    Nery da Matta, A; Silva, C B; Hassón-Voloch, A

    1996-01-01

    The effect of Mg(2+)-ATP on purified acetylcholinesterase (AChE) from electric tissue of Electrophorus electricus (L.) was studied. The enzymatic activities were measured with acetylcholine and acetylthiocholine as substrates. The kinetic parameters Vmax, Km and Hill coefficient (nH), for acetylcholine and acetylthiocholine were modified with Mg(2+)-ATP. It was shown that acetylcholinesterase presents an apparent activation at high concentration of substrates and an inhibition in the presence of Mg(2+)-ATP at low concentration of acetylcholine and acetylthiocholine. In addition, the data suggest that Mg(2+)-ATP induced an allosteric modulation of the acetylcholinesterase obtained from Electrophorous electricus (L.), and indicate an active adenosine triphosphate participation during cholinergic activity.

  5. Detection of microbial contaminations in drinking water using ATP measurements – evaluating potential for online monitoring

    DEFF Research Database (Denmark)

    Vang, Óluva Karin; Corfitzen, Charlotte B.; Albrechtsen, Hans-Jørgen

    2011-01-01

    There is an increasing call for fast and reliable methods for continuous monitoring of microbial drinking water quality in order to protect public health. The potential for Adenosine triphosphate (ATP) measurements as a real-time analysis for continuous monitoring of microbial drinking water...... quality was investigated through simulation of two contamination scenarios, i.e. drinking water contaminated with waste water and surface water at various concentrations. With ATP measurements it was possible to detect waste water diluted 1000-10,000 times in drinking water depending on sensitivity...... of reagent kit. Surface water diluted 100-1000 times was detected in drinking water with ATP measurements. ATP has the potential as an early warning tool, especially in the period when the contamination concentration is high. 2011 © American Water Works Association AWWA WQTC Conference Proceedings All Rights...

  6. Detection of microbial contaminations in drinking water using ATP measurements – evaluating potential for online monitoring

    DEFF Research Database (Denmark)

    Vang, Óluva Karin; Corfitzen, Charlotte B.; Albrechtsen, Hans-Jørgen

    2011-01-01

    There is an increasing call for fast and reliable methods for continuous monitoring of microbial drinking water quality in order to protect public health. The potential for Adenosine triphosphate (ATP) measurements as a real-time analysis for continuous monitoring of microbial drinking water...... quality was investigated through simulation of two contamination scenarios, i.e. drinking water contaminated with waste water and surface water at various concentrations. With ATP measurements it was possible to detect waste water diluted 1000-10,000 times in drinking water depending on sensitivity...... of reagent kit. Surface water diluted 100-1000 times was detected in drinking water with ATP measurements. ATP has the potential as an early warning tool, especially in the period when the contamination concentration is high. 2011 © American Water Works Association AWWA WQTC Conference Proceedings All Rights...

  7. ATP as a peripheral mediator of pain.

    Science.gov (United States)

    Hamilton, S G; McMahon, S B

    2000-07-01

    This article reviews the extent to which recent studies substantiate the hypothesis that ATP functions as a peripheral pain mediator. The discovery of the P2X family of ion channels (for which ATP is a ligand) and, in particular, the highly selective distribution of the P2X(3) receptor within the rat nociceptive system has inspired a variety of approaches to elucidate the potential role of ATP as a pain mediator. ATP elicits excitatory inward currents in small diameter sensory ganglion cells. These currents resemble those elicited by ATP on recombinantly expressed heteromeric P2X(2/3) channels as well as homomultimers consisting of P2X(2) and P2X(3). In vivo behavioural models have characterised the algogenic properties of ATP in normal conditions and in models of peripheral sensitisation. In humans, iontophoresis of ATP induces modest pain. In rats and humans the response is dependent on capsaicin sensitive neurons and is augmented in the presence of inflammatory mediators. Since ATP can be released in the vicinity of peripheral nociceptive terminals under a variety of conditions, there exists a purinergic chain of biological processes linking tissue damage to pain perception. The challenge remains to prove a physiological role for endogenous ATP in activating this chain of events.

  8. Factors Impeding Enzymatic Wheat Gluten Hydrolysis at High Solid Concentrations

    NARCIS (Netherlands)

    Hardt, N.A.; Janssen, A.E.M.; Boom, R.M.; Goot, van der A.J.

    2014-01-01

    Enzymatic wheat gluten hydrolysis at high solid concentrations is advantageous from an environmental and economic point of view. However, increased wheat gluten concentrations result in a concentration effect with a decreased hydrolysis rate at constant enzyme-to-substrate ratios and a decreased

  9. High-throughput downstream process development for cell-based products using aqueous two-phase systems (ATPS) - A case study.

    Science.gov (United States)

    Zimmermann, Sarah; Scheeder, Christian; Zimmermann, Philipp K; Bogsnes, Are; Hansson, Mattias; Staby, Arne; Hubbuch, Jürgen

    2017-02-01

    The availability of preparative-scale downstream processing strategies for cell-based products presents a critical juncture between fundamental research and clinical development. Aqueous two-phase systems (ATPS) present a gentle, scalable, label-free, and cost-effective method for cell purification, and are thus a promising tool for downstream processing of cell-based therapeutics. Here, the application of a previously developed robotic screening platform that enables high-throughput cell partitioning analysis in ATPS is reported. In the present case study a purification strategy for two model cell lines based on high-throughput screening (HTS)-data and countercurrent distribution (CCD)-modeling, and validated the CCD-model experimentally is designed. The obtained data are shown an excellent congruence between CCD-model and experimental data, indicating that CCD-models in combination with HTS-data are a powerful tool in downstream process development. Finally, the authors are shown that while cell cycle phase significantly influences cell partitioning, cell type specific differences in surface properties are the main driving force in charge-dependent separation of HL-60 and L929 cells. In order to design a highly robust purification process it is, however, advisable to maintain constant growth conditions. Copyright © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Arecoline improves vascular endothelial function in high fructose-fed rats via increasing cystathionine-γ-lyase expression and activating K(ATP) channels.

    Science.gov (United States)

    Ling, Hong-yan; Wang, Guang; Zhang, Wei; Li, Xing; Zhou, Shou-hong; Hu, Bi

    2012-08-01

    To investigate the effect of arecoline, a major component of betel nut, on vascular endothelial function in high fructose-fed rats and the potential mechanisms underlying the effect. Male Wistar rats were fed a high-fructose or control diet for 16 weeks. At the beginning of week 13, the rats were injected ip with low (0.5 mg·kg(-1)·d(-1)), medium (1.0 mg·kg(-1)·d(-1)) or high (5.0 mg·kg(-1)·d(-1)) doses of arecoline for 4 weeks. At the termination of the treatments, blood was collected, fasting blood glucose (FBG) and serum insulin (FSI) levels were measured, and insulin sensitivity index (ISI) was calculated. The thoracic aortas were isolated and aortic rings were prepared for studying ACh-induced endothelium-dependent vasorelaxation (EDVR). The mRNA and protein expression of cystathionine-γ-lyase (CSE) in the thoracic aortas was analyzed using RT-PCR and Western blot analysis, respectively. In high fructose-fed rats, the levels of FBG and FSI were remarkably increased, whereas the ISI and the mRNA and protein expression of CSE were significantly decreased. ACh-induced EDVR in the aortic rings from high fructose-fed rats was remarkably reduced. These changes were reversed by treatment with high dose arecoline. Pretreatment of the aortic rings rings from high fructose-fed rats with the CSE inhibitor propargylglycine (10 mmol/L) or the ATP-sensitive potassium (K(ATP)) channel blocker glibenclamide (10 mmol/L) abolished the restoration of ACh-induced EDVR by high dose arecoline. On the contrary, treatment with high dose arecoline significantly impaired ACh-induced EDVR in the aortic rings from control rats, and pretreatment with propargylglycine or glibenclamide did not cause further changes. Arecoline treatment improves ACh-induced EDVR in high fructose-fed rats, and the potential mechanism of action might be associated with increase of CSE expression and activation of K(ATP) channels by arecoline.

  11. 生物发光法快速检测细胞内三磷酸腺苷%Rapid Detection of ATP Concentration in Cells Using Bioluminescence Method

    Institute of Scientific and Technical Information of China (English)

    郝巧玲; 吕斌; 周宜开; 程光明

    2005-01-01

    目的建立检测细胞内三磷酸腺苷(ATP)含量的生物发光方法,并利用该方法检测红细胞和不同再灌注处理心肌细胞内ATP的含量. 方法制备所需测定的细胞ATP提取液,采用荧光素-荧光素酶检测系统检测ATP含量,ATP的含量与系统中所发射的光子数成正比,发光强度越大,ATP含量越高,通过检测发光强度计算ATP的含量.结果通过对红细胞内ATP含量的测定,确定了测定细胞内ATP的最佳反应条件:pH=7.8,温度20~25 ℃,荧光素酶浓度3.0 mg/ml .利用确定的最佳条件对各种不同处理的心肌细胞内ATP浓度进行了检测,ATP检测结果与心肌功能的实验观察结果符合. 结论生物发光方法测定ATP含量方法简单、结果可靠,可以在短时间内得到检测结果,是一种检测细胞内ATP的有效方法.

  12. Refractive Secondary Solar Concentrator Demonstrated High-Temperature Operation

    Science.gov (United States)

    Wong, Wayne A.

    2002-01-01

    Space applications that utilize solar thermal energy--such as electric power conversion systems, thermal propulsion systems, and furnaces--require highly efficient solar concentration systems. The NASA Glenn Research Center is developing the refractive secondary concentrator, which uses refraction and total internal reflection to efficiently concentrate and direct solar energy. When used in combination with advanced lightweight primary concentrators, such as inflatable thin films, the refractive secondary concentrator enables very high system concentration ratios and very high temperatures. Last year, Glenn successfully demonstrated a secondary concentrator throughput efficiency of 87 percent, with a projected efficiency of 93 percent using an antireflective coating. Building on this achievement, Glenn recently successfully demonstrated high-temperature operation of the secondary concentrator when it was used to heat a rhenium receiver to 2330 F. The high-temperature demonstration of the concentrator was conducted in Glenn's 68-ft long Tank 6 thermal vacuum facility equipped with a solar simulator. The facility has a rigid panel primary concentrator that was used to concentrate the light from the solar simulator onto the refractive secondary concentrator. NASA Marshall Space Flight Center provided a rhenium cavity, part of a solar thermal propulsion engine, to serve as the high-temperature receiver. The prototype refractive secondary concentrator, measuring 3.5 in. in diameter and 11.2 in. long, is made of single-crystal sapphire. A water-cooled splash shield absorbs spillage light outside of the 3.5-in. concentrator aperture. Multilayer foil insulation composed of tungsten, molybdenum, and niobium is used to minimize heat loss from the hightemperature receiver. A liquid-cooled canister calorimeter is used to measure the heat loss through the multilayer foil insulation.

  13. Fabrication and tolerances of optics for high concentration photovoltaics

    OpenAIRE

    Benitez Gimenez, Pablo; Miñano Dominguez, Juan Carlos; Ahmadpanaih, Hamed; Mendes Lopes, Joao; Zamora Herranz, Pablo

    2014-01-01

    High Concentration Photovoltaics (HCPV) require an optical system with high efficiency, low cost and large tolerance. We describe the particularities of the HCPV applications, which constrain the optics design and the manufacturing techonologies.

  14. P2X7 receptors contribute to the currents induced by ATP in guinea pig intestinal myenteric neurons.

    Science.gov (United States)

    Valdez-Morales, Eduardo; Guerrero-Alba, Raquel; Liñán-Rico, Andrómeda; Espinosa-Luna, Rosa; Zarazua-Guzman, Sergio; Miranda-Morales, Marcela; Montaño, Luis M; Barajas-López, Carlos

    2011-10-15

    The whole-cell configuration, several pharmacological tools, and single-cell RT-PCR were used to investigate the contribution of P2X7 subunits to the ATP-induced currents (I(ATP)) in guinea pig myenteric neurons. I(ATP) was recorded in the great majority of tested neurons. ATP concentration-response curve (0.01-10mM) showed two phases, the first mediated by high-sensitive P2X receptors (hsP2X receptors), observed between 0.01-0.3mM and the second mediated by low-sensitive P2X receptors (lsP2X receptors). The calculated EC(50) values of these phases were 38 and 1759 μM, respectively. 2'-3'-O-(4-benzoylbenzoyl)-ATP (BzATP) concentration-response curve was monophasic (0.01-1mM), and less potent (EC(50) 142 μM) than ATP to activate hsP2X receptors. A strong inward rectification was noticed when hsP2X receptors were activated with ATP (0.1mM) and for BzATP-induced currents (0.1mM; I(BzATP)) but a significant lower rectification was noticed when lsP2X receptors were activated (5mM). Brilliant blue G (BBG) at a concentration of 0.3 μM (known to inhibit only P2X7 receptors) reduced I(ATP) when lsP2X receptors contributed to it but neither affect hsP2X receptors nor I(BzATP). However, hsP2X receptors and I(BzATP) were both inhibited by concentrations ≥ 1 μM of this antagonist. BzATP inhibited hsP2X receptors and therefore, it behaves as partial agonist on these receptors. Using the single-cell RT-PCR technique P2X7 mRNA was detectable in 7 out of 13 myenteric neurons exhibiting P2X2 mRNA. Altogether, our results show that low-sensitive P2X receptors are likely P2X7, whereas, the high-sensitive P2X channels are probably constituted, at least in part, by P2X2 subunits.

  15. Highly-efficient gating of solid-state nanochannels by DNA supersandwich structure containing ATP aptamers: a nanofluidic IMPLICATION logic device.

    Science.gov (United States)

    Jiang, Yanan; Liu, Nannan; Guo, Wei; Xia, Fan; Jiang, Lei

    2012-09-19

    Integrating biological components into artificial devices establishes an interface to understand and imitate the superior functionalities of the living systems. One challenge in developing biohybrid nanosystems mimicking the gating function of the biological ion channels is to enhance the gating efficiency of the man-made systems. Herein, we demonstrate a DNA supersandwich and ATP gated nanofluidic device that exhibits high ON-OFF ratios (up to 10(6)) and a perfect electric seal at its closed state (~GΩ). The ON-OFF ratio is distinctly higher than existing chemically modified nanofluidic gating systems. The gigaohm seal is comparable with that required in ion channel electrophysiological recording and some lipid bilayer-coated nanopore sensors. The gating function is implemented by self-assembling DNA supersandwich structures into solid-state nanochannels (open-to-closed) and their disassembly through ATP-DNA binding interactions (closed-to-open). On the basis of the reversible and all-or-none electrochemical switching properties, we further achieve the IMPLICATION logic operations within the nanofluidic structures. The present biohybrid nanofluidic device translates molecular events into electrical signals and indicates a built-in signal amplification mechanism for future nanofluidic biosensing and modular DNA computing on solid-state substrates.

  16. [ATP in the metabolism of ruminants].

    Science.gov (United States)

    Bergner, H

    1991-10-01

    The ATP yield from the carbohydrates of anaerobically living microorganisms in the rumen amounts to only 5-10% of the ATP yield of the intermediary metabolism in the presence of oxygen. Vital functions and thus microbial protein synthesis are due to protein degradation in the rumen. The ATP yield in the intermediary metabolism of ruminants is mainly achieved from propionate and microbial protein by means of gluconeogenesis because the absorption of glucose from digested starch is very low. The relationships between ATP yield in the rumen and the processes of glucose provision for the production of lactose as well as the protein content of the milk are shown. As important processes of ATP production in microorganisms from easily soluble carbohydrates take place in silage preparations before feed intake, the corresponding consequences for the metabolism of high-performance cows fed with silage are shown.

  17. Modelling the ATP production in mitochondria

    CERN Document Server

    Saa, Alberto

    2012-01-01

    We revisit here the mathematical model for ATP production in mitochondria introduced recently by Bertram, Pedersen, Luciani, and Sherman (BPLS) as a simplification of the more complete but intricate Magnus and Keizer's model. We correct some inaccuracies in the BPLS original approximations and then analyze some of the dynamical properties of the model. We infer from exhaustive numerical explorations that the enhanced BPLS equations have a unique attractor fixed point for physiologically acceptable ranges of mitochondrial variables and respiration inputs. We determine, in the stationary regime, the dependence of the mitochondrial variables on the respiration inputs, namely the cytosolic concentration of calcium ${\\rm Ca}_{\\rm c}$ and the substrate fructose 1,6-bisphosphate FBP. The same effect of calcium saturation reported for the original BPLS model is observed here. We find out, however, an interesting non-stationary effect: the inertia of the model tends to increase considerably for high concentrations of ...

  18. P2X7 receptors and Fyn kinase mediate ATP-induced oligodendrocyte progenitor cell migration.

    Science.gov (United States)

    Feng, Ji-Feng; Gao, Xiao-Fei; Pu, Ying-Yan; Burnstock, Geoffrey; Xiang, Zhenghua; He, Cheng

    2015-09-01

    Recruitment of oligodendrocyte precursor cells (OPCs) to the lesions is the most important event for remyelination after central nervous system (CNS) injury or in demyelinating diseases. However, the underlying molecular mechanism is not fully understood. In the present study, we found high concentrations of ATP could increase the number of migrating OPCs in vitro, while after pretreatment with oxidized ATP (a P2X7 receptor antagonist), the promotive effect was attenuated. The promotive effect of 2'(3')-O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzATP) (a P2X7 receptor agonist) was more potent than ATP. After incubation with BzATP, the activity of Fyn, one member of the Src family of kinases, was enhanced. Moreover, the interaction between P2X7 and Fyn was identified by co-immunoprecipitation. After blocking the activity of Fyn or down-regulating the expression of Fyn, the migration of OPCs induced by BzATP was inhibited. These data indicate that P2X7 receptors/Fyn may mediate ATP-induced OPC migration under pathological conditions.

  19. Interference sources in ATP bioluminescence assay of silica nanoparticle toxicity to activated sludge.

    Science.gov (United States)

    Sibag, Mark; Kim, Seung Hwan; Kim, Choah; Kim, Hee Jun; Cho, Jinwoo

    2015-06-01

    ATP measurement provides an overview of the general state of microbial activity, and thus it has proven useful for the evaluation of nanoparticle toxicity in activated sludge. ATP bioluminescence assay, however, is susceptible to interference by the components of activated sludge other than biomass. This paper presents the interference identified specific to the use of this assay after activated sludge respiration inhibition test of silica nanoparticles (OECD 209). We observed a high degree of interference (90%) in the presence of 100 mg/L silica nanoparticles and a low level of ATP being measured (0.01 μM); and 30% interference by the synthetic medium regardless of silica nanoparticle concentration and ATP level in the samples. ATP measurement in activated sludge with different MLSS concentrations revealed interference of high biomass content. In conclusion, silica nanoparticles, synthetic medium and activated sludge samples themselves interfere with ATP bioluminescence; this will need to be considered in the evaluation of silica nanoparticle toxicity to activated sludge when this type of assay is used.

  20. Studies on luciferin-luciferase ATP assay in plants (etiolated wheat germs, and bean leaves

    Directory of Open Access Journals (Sweden)

    A. Barbaro

    2015-01-01

    Full Text Available For ATP determination by the method of bioluminescence apparatus of home production was adapted from the equipment available in any isotope laboratory. The measurement error did not exceed 1.5 per cent. Methodical experiments concerned the choice of the extraction, fixation and storage methods of plant material for determination at the given moment of the amount of ATP in the tissues, unchanged by the analytical procedure. The highest ATP amounts were recovered by extraction with perchloric acid at high (25% concentrations of the tissue in the homogenate. The best way of fixation of the material for later analyses was found to be freezing of ready extracts. Lyophilization and freezing of the plant material caused a several-fold decrease of the ATP level in the tissues. These results suggest the necessity of working in conditions of low temperature during the entire analytical procedure and strict observation of time limitation.

  1. Bile acid effects are mediated by ATP release and purinergic signalling in exocrine pancreatic cells

    DEFF Research Database (Denmark)

    Kowal, Justyna Magdalena; Haanes, Kristian Agmund; Christensen, Nynne

    2015-01-01

    of purinergic receptors. The TGR5 receptor, expressed on the luminal side of pancreatic ducts, was not involved in ATP release and Ca(2+) signals, but could stimulate Na(+)/Ca(2+) exchange in some conditions. CONCLUSIONS: CDCA evokes significant ATP release that can stimulate purinergic receptors, which in turn...... signalling are other important regulators of similar secretory mechanisms in pancreas. The aim of our study was to elucidate whether there is interplay between ATP and BA signalling. RESULTS: Here we show that CDCA (chenodeoxycholic acid) caused fast and concentration-dependent ATP release from acini (AR42J...... increase [Ca(2+)]i. The TGR5 receptor is not involved in these processes but can play a protective role at high intracellular Ca(2+) conditions. We propose that purinergic signalling could be taken into consideration in other cells/organs, and thereby potentially explain some of the multifaceted effects...

  2. Intercomparison of passive microwave sea ice concentration retrievals over the high-concentration Arctic sea ice

    DEFF Research Database (Denmark)

    andersen, susanne; Tonboe, R.; Kaleschke, L.

    2007-01-01

    [1] Measurements of sea ice concentration from the Special Sensor Microwave Imager (SSM/I) using seven different algorithms are compared to ship observations, sea ice divergence estimates from the Radarsat Geophysical Processor System, and ice and water surface type classification of 59 wide......-swath synthetic aperture radar (SAR) scenes. The analysis is confined to the high-concentration Arctic sea ice, where the ice cover is near 100%. During winter the results indicate that the variability of the SSM/I concentration estimates is larger than the true variability of ice concentration. Results from...... a trusted subset of the SAR scenes across the central Arctic allow the separation of the ice concentration uncertainty due to emissivity variations and sensor noise from other error sources during the winter of 2003-2004. Depending on the algorithm, error standard deviations from 2.5 to 5.0% are found...

  3. High Stokes shift perylene dyes for luminescent solar concentrators.

    Science.gov (United States)

    Sanguineti, Alessandro; Sassi, Mauro; Turrisi, Riccardo; Ruffo, Riccardo; Vaccaro, Gianfranco; Meinardi, Francesco; Beverina, Luca

    2013-02-25

    Highly efficient plastic based single layer Luminescent Solar Concentrators (LSCs) require the design of luminophores having complete spectral separation between absorption and emission spectra (large Stokes shift). We describe the design, synthesis and characterization of a new perylene dye possessing Stokes shift as high as 300 meV, fluorescent quantum yield in the LSC slab of 70% and high chemical and photochemical stability.

  4. Hair and toenail arsenic concentrations of residents living in areas with high environmental arsenic concentrations.

    OpenAIRE

    Hinwood, Andrea L; Sim, Malcolm R; Jolley, Damien; de Klerk, Nick; Bastone, Elisa B; Gerostamoulos, Jim; Drummer, Olaf H

    2003-01-01

    Surface soil and groundwater in Australia have been found to contain high concentrations of arsenic. The relative importance of long-term human exposure to these sources has not been established. Several studies have investigated long-term exposure to environmental arsenic concentrations using hair and toenails as the measure of exposure. Few have compared the difference in these measures of environmental sources of exposure. In this study we aimed to investigate risk factors for elevated hai...

  5. Applications of nonimaging optics for very high solar concentrations

    Energy Technology Data Exchange (ETDEWEB)

    O`Gallagher, J.; Winston, R.

    1997-12-31

    Using the principles and techniques of nonimaging optics, solar concentrations that approach the theoretical maximum can be achieved. This has applications in solar energy collection wherever concentration is desired. In this paper, we survey recent progress in attaining and using high and ultrahigh solar fluxes. We review a number of potential applications for highly concentrated solar energy and the current status of the associated technology. By making possible new and unique applications for intense solar flux, these techniques have opened a whole new frontier for research and development of potentially economic uses of solar energy.

  6. Electron beam treatment of exhaust gas with high NOx concentration

    Science.gov (United States)

    Licki, Janusz; Chmielewski, Andrzej G.; Pawelec, Andrzej; Zimek, Zbigniew; Witman, Sylwia

    2014-05-01

    Simulated exhaust gases with a high NOx concentration, ranging from 200 to 1700 ppmv, were irradiated by an electron beam from an accelerator. In the first part of this study, only exhaust gases were treated. Low NOx removal efficiencies were obtained for high NOx concentrations, even with high irradiation doses applied. In the second part of study, gaseous ammonia or/and vapor ethanol were added to the exhaust gas before its inlet to the plasma reactor. These additions significantly enhanced the NOx removal efficiency. The synergistic effect of high SO2 concentration on NOx removal was observed. The combination of electron beam treatment with the introduction of the above additions and with the performance of irradiation under optimal parameters ensured high NOx removal efficiency without the application of a solid-state catalyst.

  7. [Reasons of high concentration ammonium in Yellow River, China].

    Science.gov (United States)

    Zhang, Xue-qing; Xia, Xing-hui; Yang, Zhi-feng

    2007-07-01

    Ammonium nitrogen contamination is one of the major problems of the Yellow River in China. The speciation, concentration and sources of nitrogen compounds as well as the water environment conditions of the Yellow River had been analyzed to study the reasons for the fact that the ammonium nitrogen concentration was above the water quality standard. In addition, laboratory experiments had been carried out to investigate the effects of suspended sediment (SS) on nitrification rate. The results indicated that the presence of SS could accelerate the nitrification process, therefore, the effects of SS on nitrification rate was not the reason for the high level of ammonium nitrogen in the river. The excessive and continuous input of nitrogen contaminants to the river was the fundamental reason for the high concentration of ammonium nitrogen. Organic and ammonium nitrogen with high concentration inhibitted the nitrification processes. When the initial NH4+ -N concentrations were 10.1, 18.4 and 28.2 mg/L, nitrification efficiencies were 17.4%, 13.0% and 2.5%, respectively. When the initial organic nitrogen concentrations were 5.5 and 8.6 mg/L, the maximum concentrations of ammonium nitrogen produced by the oxidation of organic nitrogen would reach 0.47 and 1.69 mg/L and they would last for 2 days and 6 days, respectively. The oxygen-consuming organics and toxic substance existing in the river water could inhibit the activity of nitrifying bacteria, and thus lead to the accumulation of ammonium nitrogen. In addition, the high pH value of river water resulted in the high concentration of nonionic ammonium nitrogen which would reduce the activity of nitrifying bacteria and decrease the nitrification rates. Besides, low river runoff, low SS content and low activity of nitrifying bacteria resulted in the high level of ammonium nitrogen of the river in the low water season.

  8. Differences in Swallowing between High and Low Concentration Taste Stimuli

    Directory of Open Access Journals (Sweden)

    Ahmed Nagy

    2014-01-01

    Full Text Available Taste is a property that is thought to potentially modulate swallowing behavior. Whether such effects depend on taste, intensity remains unclear. This study explored differences in the amplitudes of tongue-palate pressures in swallowing as a function of taste stimulus concentration. Tongue-palate pressures were collected in 80 healthy women, in two age groups (under 40, over 60, stratified by genetic taste status (nontasters, supertasters. Liquids with different taste qualities (sweet, sour, salty, and bitter were presented in high and low concentrations. General labeled magnitude scale ratings captured perceived taste intensity and liking/disliking of the test liquids. Path analysis explored whether factors of taste, concentration, age group, and/or genetic taste status impacted: (1 perceived intensity; (2 palatability; and (3 swallowing pressures. Higher ratings of perceived intensity were found in supertasters and with higher concentrations, which were more liked/disliked than lower concentrations. Sweet stimuli were more palatable than sour, salty, or bitter stimuli. Higher concentrations elicited stronger tongue-palate pressures independently and in association with intensity ratings. The perceived intensity of a taste stimulus varies as a function of stimulus concentration, taste quality, participant age, and genetic taste status and influences swallowing pressure amplitudes. High-concentration salty and sour stimuli elicit the greatest tongue-palate pressures.

  9. High Black Carbon (BC) Concentrations along Indian National Highways

    Science.gov (United States)

    Kumar, S.; Singh, A. K.; Singh, R. P.

    2015-12-01

    Abstract:Black carbon (BC), the optically absorbing component of carbonaceous aerosol, has direct influence on radiation budget and global warming. Vehicular pollution is one of the main sources for poor air quality and also atmospheric pollution. The number of diesel vehicles has increased on the Indian National Highways during day and night; these vehicles are used for the transport of goods from one city to another city and also used for public transport. A smoke plume from the vehicles is a common feature on the highways. We have made measurements of BC mass concentrations along the Indian National Highways using a potable Aethalometer installed in a moving car. We have carried out measurements along Varanasi to Kanpur (NH-2), Varanasi to Durgapur (NH-2), Varanasi to Singrauli (SH-5A) and Varanasi to Ghazipur (NH-29). We have found high concentration of BC along highways, the average BC mass concentrations vary in the range 20 - 40 µg/m3 and found high BC mass concentrations up to 600 μg/m3. Along the highways high BC concentrations were characteristics of the presence of industrial area, power plants, brick kilns and slow or standing vehicles. The effect of increasing BC concentrations along the National Highways and its impact on the vegetation and human health will be presented. Key Words: Black Carbon; Aethalometer; mass concentration; Indian National Highways.

  10. Estimation of PMI depends on the changes in ATP and its degradation products.

    Science.gov (United States)

    Mao, Shiwei; Fu, Gaowen; Seese, Ronald R; Wang, Zhen-Yuan

    2013-09-01

    Estimating the time since death, or postmortem interval (PMI), has been one of the biggest difficulties in modern forensic investigation. This study tests if the concentrations of breakdown products of adenosine triphosphate (ATP) correlate with PMI in multiple organs from rat. Brains, spleens, and kidneys of rats were harvested at different time points in carcasses maintained at 4°C or 20°C. High Performance Liquid Chromatography (HPLC) was used to quantify concentrations of metabolites related to ATP degradation. A K value (Kv=100×(Hx+HxR)/(ATP+ADP+AMP+IMP+HxR+Hx)) was calculated and correlated with PMI for each organ and temperature. The results indicate that the K value is a robust index for the estimation of PMI based on highly significant linear correlations between PMI and concentrations of ATP breakdown products. Compared with other current research methods, the changing tendency of ATP and its degradation products may be potentially a better way for the estimation of PMI in medico-legal practice. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  11. Real-time luminescence imaging of cellular ATP release.

    Science.gov (United States)

    Furuya, Kishio; Sokabe, Masahiro; Grygorczyk, Ryszard

    2014-03-15

    Extracellular ATP and other purines are ubiquitous mediators of local intercellular signaling within the body. While the last two decades have witnessed enormous progress in uncovering and characterizing purinergic receptors and extracellular enzymes controlling purinergic signals, our understanding of the initiating step in this cascade, i.e., ATP release, is still obscure. Imaging of extracellular ATP by luciferin-luciferase bioluminescence offers the advantage of studying ATP release and distribution dynamics in real time. However, low-light signal generated by bioluminescence reactions remains the major obstacle to imaging such rapid processes, imposing substantial constraints on its spatial and temporal resolution. We have developed an improved microscopy system for real-time ATP imaging, which detects ATP-dependent luciferin-luciferase luminescence at ∼10 frames/s, sufficient to follow rapid ATP release with sensitivity of ∼10 nM and dynamic range up to 100 μM. In addition, simultaneous differential interference contrast cell images are acquired with infra-red optics. Our imaging method: (1) identifies ATP-releasing cells or sites, (2) determines absolute ATP concentration and its spreading manner at release sites, and (3) permits analysis of ATP release kinetics from single cells. We provide instrumental details of our approach and give several examples of ATP-release imaging at cellular and tissue levels, to illustrate its potential utility.

  12. Customized ATP towpreg. [Automated Tow Placement

    Science.gov (United States)

    Sandusky, Donald A.; Marchello, Joseph M.; Baucom, Robert M.; Johnston, Norman J.

    1992-01-01

    Automated tow placement (ATP) utilizes robotic technology to lay down adjacent polymer-matrix-impregnated carbon fiber tows on a tool surface. Consolidation and cure during ATP requires that void elimination and polymer matrix adhesion be accomplished in the short period of heating and pressure rolling that follows towpreg ribbon placement from the robot head to the tool. This study examined the key towpreg ribbon properties and dimensions which play a significant role in ATP. Analysis of the heat transfer process window indicates that adequate heating can be achieved at lay down rates as high as 1 m/sec. While heat transfer did not appear to be the limiting factor, resin flow and fiber movement into tow lap gaps could be. Accordingly, consideration was given to towpreg ribbon having uniform yet non-rectangular cross sections. Dimensional integrity of the towpreg ribbon combined with customized ribbon architecture offer great promise for processing advances in ATP of high performance composites.

  13. An RNA motif that binds ATP

    Science.gov (United States)

    Sassanfar, M.; Szostak, J. W.

    1993-01-01

    RNAs that contain specific high-affinity binding sites for small molecule ligands immobilized on a solid support are present at a frequency of roughly one in 10(10)-10(11) in pools of random sequence RNA molecules. Here we describe a new in vitro selection procedure designed to ensure the isolation of RNAs that bind the ligand of interest in solution as well as on a solid support. We have used this method to isolate a remarkably small RNA motif that binds ATP, a substrate in numerous biological reactions and the universal biological high-energy intermediate. The selected ATP-binding RNAs contain a consensus sequence, embedded in a common secondary structure. The binding properties of ATP analogues and modified RNAs show that the binding interaction is characterized by a large number of close contacts between the ATP and RNA, and by a change in the conformation of the RNA.

  14. Sterile Filtration of Highly Concentrated Protein Formulations: Impact of Protein Concentration, Formulation Composition, and Filter Material.

    Science.gov (United States)

    Allmendinger, Andrea; Mueller, Robert; Huwyler, Joerg; Mahler, Hanns-Christian; Fischer, Stefan

    2015-10-01

    Differences in filtration behavior of concentrated protein formulations were observed during aseptic drug product manufacturing of biologics dependent on formulation composition. The present study investigates filtration forces of monoclonal antibody formulations in a small-scale set-up using polyvinylidene difluoride (PVDF) or polyethersulfone (PES) filters. Different factors like formulation composition and protein concentration related to differences in viscosity, as well as different filtration rates were evaluated. The present study showed that filtration behavior was influenced by the presence or absence of a surfactant in the formulation, which defines the interaction between filter membrane and surface active formulation components. This can lead to a change in filter resistance (PES filter) independent on the buffer system used. Filtration behavior was additionally defined by rheological non-Newtonian flow behavior. The data showed that high shear rates resulting from small pore sizes and filtration pressure up to 1.0 bar led to shear-thinning behavior for highly concentrated protein formulations. Differences in non-Newtonian behavior were attributed to ionic strength related to differences in repulsive and attractive interactions. The present study showed that the interplay of formulation composition, filter material, and filtration rate can explain differences in filtration behavior/filtration flux observed for highly concentrated protein formulations thus guiding filter selection.

  15. The action of selenite on ATP synthesis in rat lens

    OpenAIRE

    Adamchak, Marsha Ann

    1986-01-01

    A subcutaneous injection of sodium selenite (30 umol/kg body weight) in 10â day old rats produced a cataract within 72 hours. Lens opacification was preceded by a 15% decrease in ATP content. Lens ATP did not fully recover to control concentrations by 11 days postâ injection. A moderate correlation existed between lens weight and total ATP content in control lenses

  16. Adsorption of nucleotides on biomimetic apatite: The case of adenosine 5⿲ triphosphate (ATP)

    Science.gov (United States)

    Hammami, Khaled; El-Feki, Hafed; Marsan, Olivier; Drouet, Christophe

    2016-01-01

    ATP is a well-known energy supplier in cells. The idea to associate ATP to pharmaceutical formulations/biotechnological devices to promote cells activity by potentially modulating their microenvironment thus appears as an appealing novel approach. Since biomimetic nanocrystalline apatites have shown great promise for biomedical applications (bone regeneration, cells diagnostics/therapeutics, ⿦), thanks to a high surface reactivity and an intrinsically high biocompatibility, the present contribution was aimed at exploring ATP/apatite interactions. ATP adsorption on a synthetic carbonated nanocrystalline apatite preliminarily characterized (by XRD, FTIR, Raman, TG-DTA and SEM-EDX) was investigated in detail, pointing out a good agreement with Sips isothermal features. Adsorption characteristics were compared to those previously obtained on monophosphate nucleotides (AMP, CMP), unveiling some specificities. ATP was found to adsorb effectively onto biomimetic apatite: despite smaller values of the affinity constant KS and the exponential factor m, larger adsorbed amounts were reached for ATP as compared to AMP for any given concentration in solution. m guided by direct surface bonding rather than through stabilizing intermolecular interactions. Although standard οGads ° was estimated to only ⿿4 kJ/mol, the large value of Nmax led to significantly negative effective οGads values down to ⿿33 kJ/mol, reflecting the spontaneous character of adsorption process. Vibrational spectroscopy data (FTIR and Raman) pointed out spectral modifications upon adsorption, confirming chemical-like interactions where both the triphosphate group of ATP and its nucleic base were involved. The present study is intended to serve as a basis for future research works involving ATP and apatite nanocrystals/nanoparticles in view of biomedical applications (e.g. bone tissue engineering, intracellular drug delivery, ⿦).

  17. Denitrification of fertilizer wastewater at high chloride concentration

    DEFF Research Database (Denmark)

    Ucisik, Ahmed Süheyl; Henze, Mogens

    g/l. The results of the experiments showed that biological denitrification was feasible at the extreme environmental conditions prevailing in fertilizer wastewater. Stable continuous biological denitrfication of the synthetic high chloride wastewater was performed up to 77.4 g Cl/l at 37 degree C......Wastewater from fertilizer industry is characterized by high contents of chloride concentration, which normally vary between 60 and 76 g/l. Experiments with bilogical denitrification were performed in lab-scale "fill and draw" reactors with synthetic wastewater with chloride concentrations up to 77.4...

  18. High estrogen concentrations in receiving river discharge from a concentrated livestock feedlot.

    Science.gov (United States)

    Chen, Te-San; Chen, Ting-Chien; Yeh, Kuei-Jyum C; Chao, How-Ran; Liaw, Ean-Tun; Hsieh, Chi-Ying; Chen, Kuan-Chung; Hsieh, Lien-Te; Yeh, Yi-Lung

    2010-07-15

    Environmental estrogenic chemicals interrupt endocrine systems and generate reproductive abnormalities in wildlife, especially natural and synthetic estrogenic steroid hormones such as 17beta-estradiol (E2), estrone (E1), estriol (E3), 17alpha-ethynylestradiol (EE2), and diethylstilbestrol (DES). Concentrated animal feedlot operations (CAFOs) are of particular concern since large amounts of naturally excreted estrogens are discharged into aquatic environments. This study investigated E2, E1, E3, EE2, and DES with high performance liquid chromatography/tandem mass (HPLC-MS/MS) analyses along Wulo Creek in southern Taiwan, near a concentrated livestock feedlot containing 1,030,000 broiler chickens, 934,000 laying hens, 85,000 pigs, and 1500 cattle. Sampling was performed from December 2008 to May 2009, in which 54 samples were collected. Experimental results indicate that concentrations of EE2 were lower than the limit of detection (LOD), and concentrations of DES were only detected twice. Concentrations ranged from 7.4 to 1267 ng/L for E1, from not detected (ND) to 313.6 ng/L for E2, and from ND to 210 ng/L for E3. E1 had the highest average mass fraction (72.2 + or - 3.6%), which was significantly higher than E3 (16.2 + or - 1.7%) and E2 (11.5 + or - 2.6%). Additionally, the mean E2 equivalent quotient (EEQ) ranged from 17.3 to 137.9 ng-E2/L. Despite having a markedly lower concentration than E1, E2 more significantly contributed (52.4 + or - 6.0%) EEQ than E1 (19.7 + or - 3.5%). Moreover, the concentrations of E2, E1, and E3 upstream were significantly higher than concentrations downstream, suggesting a high attenuation effect and fast degradation in the study water. Most concentrations in winter season were higher than those of spring season due to the low dilution effect and low microbial activity in the winter season. Based on the results of this study, we recommend further treatment of the wastewater discharge from the feedlot.

  19. Biodegradation dynamics of high catechol concentrations by Aspergillus awamori.

    Science.gov (United States)

    Stanchev, Veselin; Stoilova, Ivanka; Krastanov, Albert

    2008-06-15

    The biodegradation process of high catechol concentrations by Aspergillus awamori was investigated. The values of the kinetic constants for a model of specific growth rate at different initial conditions were determined. At 1.0 g/L catechol concentration, the biodegradation process proceeded in the conditions of substrate limitation. At higher catechol concentrations (2.0 and 3.0 g/L) a presence of substrate inhibition was established. The dynamics of the specific catechol degradation rate was studied and the values of catechol and biomass concentrations, maximizing the specific catechol degradation rate, were estimated analytically. The specified ratio catechol/biomass could serve as a starting base for determination of the initial conditions for a batch process, for specifying the moment of feeding for a fed-batch process, and for monitoring and control of a continuous process by the aim of time-optimal control.

  20. High shear treatment of concentrates and drying conditions influence the solubility of milk protein concentrate powders.

    Science.gov (United States)

    Augustin, Mary Ann; Sanguansri, Peerasak; Williams, Roderick; Andrews, Helen

    2012-11-01

    The solubility of milk protein concentrate (MPC) powders was influenced by the method used for preparing the concentrate, drying conditions, and the type of dryer used. Increasing total solids of the ultrafiltered concentrates (23% total solids, TS) by diafiltration to 25% TS or evaporation to 31% TS decreased the solubility of MPC powders (80-83% protein, w/w dry basis), with ultrafiltration followed by evaporation to higher total solids having the greater detrimental effect on solubility. High shear treatment (homogenisation at 350/100 bar, microfluidisation at 800 bar or ultrasonication at 24 kHz, 600 watts) of ultrafiltered and diafiltered milk protein concentrates prior to spray drying increased the nitrogen solubility of MPC powders (82% protein, w/w dry basis). Of the treatments applied, microfluidisation was the most effective for increasing nitrogen solubility of MPC powders after manufacture and during storage. Manufacture of MPC powders (91% protein, w/w dry basis) prepared on two different pilot-scale dryers (single stage or two stage) from milk protein concentrates (20% TS) resulted in powders with different nitrogen solubility and an altered response to the effects of microfluidisation. Microfluidisation (400, 800 and 1200 bar) of the concentrate prior to drying resulted in increased long term solubility of MPC powders that were prepared on a single stage dryer but not those produced on a two stage spray dryer. This work demonstrates that microfluidisation can be used as a physical intervention for improving MPC powder solubility. Interactions between the method of preparation and treatment of concentrate prior to drying, the drying conditions and dryer type all influence MPC solubility characteristics.

  1. High-concentration planar microtracking photovoltaic system exceeding 30% efficiency

    Science.gov (United States)

    Price, Jared S.; Grede, Alex J.; Wang, Baomin; Lipski, Michael V.; Fisher, Brent; Lee, Kyu-Tae; He, Junwen; Brulo, Gregory S.; Ma, Xiaokun; Burroughs, Scott; Rahn, Christopher D.; Nuzzo, Ralph G.; Rogers, John A.; Giebink, Noel C.

    2017-08-01

    Prospects for concentrating photovoltaic (CPV) power are growing as the market increasingly values high power conversion efficiency to leverage now-dominant balance of system and soft costs. This trend is particularly acute for rooftop photovoltaic power, where delivering the high efficiency of traditional CPV in the form factor of a standard rooftop photovoltaic panel could be transformative. Here, we demonstrate a fully automated planar microtracking CPV system solar cell at >660× concentration ratio over a 140∘ full field of view. In outdoor testing over the course of two sunny days, the system operates automatically from sunrise to sunset, outperforming a 17%-efficient commercial silicon solar cell by generating >50% more energy per unit area per day in a direct head-to-head competition. These results support the technical feasibility of planar microtracking CPV to deliver a step change in the efficiency of rooftop solar panels at a commercially relevant concentration ratio.

  2. Bioluminescence microscopy: application to ATP measurements in single living cells

    Science.gov (United States)

    Brau, Frederic; Helle, Pierre; Bernengo, Jean C.

    1997-12-01

    Bioluminescence microscopy can be used to measure intracellular cofactors and ionic concentrations (Ca2+, K+, ATP, NADH), as an alternative to micro- spectrophotometry and micro-fluorimetry, due to the development of sensitive detectors (cooled photomultipliers tubes and CCD). The main limitation comes from the very small and brief intensity of the emitted light. Our instrumentation based on an inverted microscope, equipped with high aperture immersion lenses is presented. Light intensity measurements are carried out through a photomultiplier sorted for low dark current and cooled at -5 degree(s)C to reduce thermal noise. Our first aim is to quantify ATP on single living cells using the firefly luciferin-luciferase couple. Experimental and kinetic aspects are presented to emphasize the potentialities of the technique.

  3. Beryllium-10 concentrations in water samples of high northern latitudes

    Energy Technology Data Exchange (ETDEWEB)

    Strobl, C.; Eisenhauer, A.; Schulz, V.; Baumann, S.; Mangini, A. [Heidelberger Akademie der Wissenschaften, Heildelberg (Germany); Kubik, P.W. [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

    1997-09-01

    {sup 10}Be concentrations in the water column of high northern latitudes were not available so far. We present different {sup 10}Be profiles from the Norwegian-Greenland Sea, the Arctic Ocean, and the Laptev Sea. (author) 3 fig., 3 refs.

  4. Optimization of Scatterer Concentration in High-Gain Scattering Media

    Institute of Scientific and Technical Information of China (English)

    ZHU Jiu-Gao; ZHU He-Yuan; SUN Die-Chi; DU Ge-Guo; LI Fu-Ming

    2001-01-01

    We report the scatterer concentration-dependent behaviour of laser action in high-gain scattering media. Amodified model of a random laser is proposed to explain the experimental results in good agreement. We mayuse this modified model to design and optimize the random laser system. A further detailed model is needed toquantitatively analyse the far-field distribution of random laser action.

  5. [Toxic effects of high concentrations of ammonia on Euglena gracilis].

    Science.gov (United States)

    Liu, Yan; Shi, Xiao-Rong; Cui, Yi-Bin; Li, Mei

    2013-11-01

    Ammonia is among the common contaminants in aquatic environments. The present study aimed at evaluation of the toxicity of ammonia at high concentration by detecting its effects on the growth, pigment contents, antioxidant enzyme activities, and DNA damage (comet assay) of a unicellular microalga, Euglena gracilis. Ammonia restrained the growth of E. gracilis, while at higher concentrations, ammonia showed notable inhibition effect, the growth at 2 000 mg x L(-1) was restrained to 55.7% compared with that of the control; The contents of photosynthetic pigments and protein went up with increasing ammonia dosage and decreased when the ammonia concentration was above 1000 mg x L(-1); In addition, there was an obvious increase in SOD and POD activities, at higher concentration (2 000 mg x L(-1)), activities of SOD and POD increased by 30.7% and 49.4% compared with those of the control, indicating that ammonia could promote activities of antioxidant enzymes in E. gracilis; The degree of DNA damage observed in the comet assay increased with increasing ammonia concentration, which suggested that high dose of ammonia may have potential mutagenicity on E. gracilis.

  6. High-efficiency organic solar concentrators for photovoltaics.

    Science.gov (United States)

    Currie, Michael J; Mapel, Jonathan K; Heidel, Timothy D; Goffri, Shalom; Baldo, Marc A

    2008-07-11

    The cost of photovoltaic power can be reduced with organic solar concentrators. These are planar waveguides with a thin-film organic coating on the face and inorganic solar cells attached to the edges. Light is absorbed by the coating and reemitted into waveguide modes for collection by the solar cells. We report single- and tandem-waveguide organic solar concentrators with quantum efficiencies exceeding 50% and projected power conversion efficiencies as high as 6.8%. The exploitation of near-field energy transfer, solid-state solvation, and phosphorescence enables 10-fold increases in the power obtained from photovoltaic cells, without the need for solar tracking.

  7. Bioleaching of marmatite in high concentration of iron

    Institute of Scientific and Technical Information of China (English)

    邱冠周; 吴伯增; 覃文庆; 蓝卓越

    2002-01-01

    Bioleaching of marmatite with a culture of Thiobacillus ferrooxidans and Thiobacillus thiooxidans in high concentration of iron was studied, the results show that the zinc leaching rate of the mixed culture is faster than that of the sole Thiobacillus ferrooxidans, the increasing iron concentration in leaching solution enhances the zinc leaching rate. The SEM analysis indicates that the chemical leaching residues is covered with porous solid layer of elemental sulfur, while elemental sulfur is not found in the bacterial leaching residues. The primary role of bacteria in bioleaching of sphalerite is to oxidize the chemical leaching products of ferrous ion and elemental sulfur, thus the indirect mechanism prevails in the bioleaching of marmatite.

  8. High concentration of calcium ions in Golgi apparatus

    Institute of Scientific and Technical Information of China (English)

    XUESHAOBAI; M.ROBERTNICOUD; 等

    1994-01-01

    The interphase NIH3T3 cells were vitally fluorescentstained with calcium indicator fluo-3 and Glogi probe C6-NBD-ceramide,and then the single cells were examined by laser scanning confocal microscopy(LSCFM) for subcellular distributions of Ca2+ and the location of Golgi apparatus.In these cells,the intracellular Ca2+ were found to be highly concentrated in the Golgi apparatus.The changes of distribution of cytosolic high Ca2+ region and the Golgi apparatus coincided with the cell cycle phase.In calcium free medium,when the plasma membrane of the cells which had been loaded with fluo-3/AM were permeated by digitonin,the fluorescence of the Golgi region decreased far less than that of the cytosol.Our results indicated that the Glogi lumen retained significantly high concentration of free calcium.

  9. Review of silicon solar cells for high concentrations

    Science.gov (United States)

    Schwartz, R. J.

    1982-06-01

    The factors that limit the performance of high concentration silicon solar cells are reviewed. The design of a conventional high concentration cell is discussed, together with the present state of the art. Unconventional cell designs that have been proposed to overcome the limitations of the conventional design are reviewed and compared. The current status of unconventional cells is reviewed. Among the unconventional cells discussed are the interdigitated back-contact cell, the double-sided cell, the polka dot cell, and the V-groove cell. It is noted that all the designs for unconventional cells require long diffusion lengths for high efficiency operation, even though the demands in this respect are less for those cells with the optical path longer than the diffusion path.

  10. Evidence for the Synthesis of ATP by an F0F1 ATP Synthase in Membrane Vesicles from Halorubrum Saccharovorum

    Science.gov (United States)

    Faguy, David; Lawson, Darion; Hochstein, Lawrence I.; Chang, Sherwood (Technical Monitor)

    1996-01-01

    Vesicles prepared in a buffer containing ADP, Mg(2+) and Pi synthesized ATP at an initial rate of 2 nmols/min/mg protein after acidification of the bulk medium (pH 8 (right arrow) 4). The intravesicular ATP concentration reached a steady state after about 30 seconds and slowly declined thereafter. ATP synthesis was inhibited by low concentrations of dicyclohexylcarbodiimide and m-chlorophenylhydrazone indicating that synthesis took place in response to the proton gradient. NEM and PCMS, which inhibit vacuolar ATPases and the vacuolar-like ATPases of extreme halophiles, did not affect ATP synthesis, and, in fact, produced higher steady state levels of ATP. This suggested that two ATPase activities were present, one which catalyzed ATP synthesis and one that caused its hydrolysis. Azide, a specific inhibitor of F0F1 ATP Synthases, inhibited halobacterial ATP synthesis. The distribution of acridine orange as imposed by a delta pH demonstrated that azide inhibition was not due to the collapse of the proton gradient due to azide acting as a protonophore. Such an effect was observed, but only at azide concentrations higher than those that inhibited ATP synthesis. These results confirm the earler observations with cells of H. saccharovorum and other extreme halophiles that ATP synthesis is inconsistent with the operation of a vacuolar-like ATPase. Therefore, the observation that a vacuolar-like enzyme is responsible for ATP synthesis (and which serves as the basis for imputing ATP synthesis to the vacuolar-like ATPases of the extreme halophiles, and the Archaea in general) should be taken with some degree of caution.

  11. Firefly bioluminescent assay of ATP in the presence of ATP extractant by using liposomes.

    Science.gov (United States)

    Kamidate, Tamio; Yanashita, Kenji; Tani, Hirofumi; Ishida, Akihiko; Notani, Mizuyo

    2006-01-01

    Liposomes containing phosphatidylcholine (PC) and cholesterol (Chol) were applied to the enhancer for firefly bioluminescence (BL) assay for ATP in the presence of cationic surfactants using as an extractant for the release of ATP from living cells. Benzalkonium chloride (BAC) was used as an ATP extractant. However, BAC seriously inhibited the activity of luciferase, thus resulting in the remarkable decrease in the sensitivity of the BL assay for ATP. On the other hand, we found that BAC was associated with liposomes to form cationic liposomes containing BAC. The association rate of BAC with liposomes was faster than that of BAC with luciferase. As a result, the inhibitory effect of BAC on luciferase was eliminated in the presence of liposomes. In addition, cationic liposomes thus formed enhanced BL emission. BL measurement conditions were optimized in terms of liposome charge type, liposome size, and total concentration of PC and Chol. ATP can be sensitively determined without dilution of analytical samples by using liposomes. The detection limit of ATP with and without liposomes was 100 amol and 25 fmol in aqueous ATP standard solutions containing 0.06% BAC, respectively. The method was applied to the determination of ATP in Escherichia coli extracts. The BL intensity was linear from 4 x 10(4) to 1 x 10(7) cells mL(-1) in the absence of liposomes. On the other hand, the BL intensity was linear from 4 x 10(3) to 4 x 10(6) cells mL(-1) in the presence of liposomes. The detection limit of ATP in E. coli extracts was improved by a factor of 10 via use of liposomes.

  12. Decomposition of high concentration SF6 using an electron beam

    Science.gov (United States)

    Son, Youn-Suk; Lee, Sung-Joo; Choi, Chang Yong; Park, Jun-Hyeong; Kim, Tak-Hyun; Jung, In-Ha

    2016-07-01

    In this study, high concentration SF6 (2-10%) was decomposed using an electron beam irradiation. Various influential factors were investigated to improve the destruction and removal efficiency (DRE) of SF6. The initial concentrations of SF6, absorbed doses, SF6/H2 ratios and retention times were the main factors of concern. As a result, the DRE increased as the adsorbed dose and retention time increased. The DRE of SF6 also increased up to 20% approximately when H2 was added to the reaction mixture. On the other hand, the DRE of SF6 decreased as initial concentrations of SF6 increased. Finally, the main by-product formed from SF6 decomposition by the electron beam was HF.

  13. Life at high salt concentrations, intracellular KCl concentrations, and acidic proteomes

    Directory of Open Access Journals (Sweden)

    Aharon eOren

    2013-11-01

    Full Text Available Extremely halophilic microorganisms that accumulate KCl for osmotic balance (the Halobacteriaceae, Salinibacter have a large excess of acidic amino acids in their proteins. This minireview explores the occurrence of acidic proteomes in halophiles of different physiology and phylogenetic affiliation. For fermentative bacteria of the order Halanaerobiales, known to accumulate KCl, an acidic proteome was predicted. However, this is not confirmed by genome analysis. The reported excess of acidic amino acids is due to a high content of Gln and Asn, which yield Glu and Asp upon acid hydrolysis. The closely related Halorhodospira halophila and Halorhodospira halochloris use different strategies to cope with high salt. The first has an acidic proteome and accumulates high KCl concentrations at high salt concentrations; the second does not accumulate KCl and lacks an acidic proteome. Acidic proteomes can be predicted from the genomes of some moderately halophilic aerobes that accumulate organic osmotic solutes (Halomonas elongata, Chromohalobacter salexigens and some marine bacteria. Based on the information on cultured species it is possible to understand the pI profiles predicted from metagenomic data from hypersaline environments.

  14. Bioanalytical Applications of Real-Time ATP Imaging Via Bioluminescence

    Energy Technology Data Exchange (ETDEWEB)

    Jason Alan Gruenhagen

    2003-12-12

    S monocrystals. Aggregates of nanospheres were bathed in imaging solution, and ATP bioluminescence was monitored to investigated the release kinetics of the nanosphere drug delivery systems. Addition of disulfide bond-cleaving molecules induced uncapping of the nanospheres and subsequently, the release of ATP. Increasing the concentration of the uncapping molecule decreased the temporal maximum and increased the magnitude of release of encapsulated ATP from the nanospheres. Furthermore, the release kinetics from the nanospheres varied with the size of the particle aggregates.

  15. The ATP Receptors P2X7 and P2X4 Modulate High Glucose and Palmitate-Induced Inflammatory Responses in Endothelial Cells.

    Directory of Open Access Journals (Sweden)

    Ramasri Sathanoori

    Full Text Available Endothelial cells lining the blood vessels are principal players in vascular inflammatory responses. Dysregulation of endothelial cell function caused by hyperglycemia, dyslipidemia, and hyperinsulinemia often result in impaired vasoregulation, oxidative stress, inflammation, and altered barrier function. Various stressors including high glucose stimulate the release of nucleotides thus initiating signaling via purinergic receptors. However, purinergic modulation of inflammatory responses in endothelial cells caused by high glucose and palmitate remains unclear. In the present study, we investigated whether the effect of high glucose and palmitate is mediated by P2X7 and P2X4 and if they play a role in endothelial cell dysfunction. Transcript and protein levels of inflammatory genes as well as reactive oxygen species production, endothelial-leukocyte adhesion, and cell permeability were investigated in human umbilical vein endothelial cells exposed to high glucose and palmitate. We report high glucose and palmitate to increase levels of extracellular ATP, expression of P2X7 and P2X4, and inflammatory markers. Both P2X7 and P2X4 antagonists inhibited high glucose and palmitate-induced interleukin-6 levels with the former having a significant effect on interleukin-8 and cyclooxygenase-2. The effect of the antagonists was confirmed with siRNA knockdown of the receptors. In addition, P2X7 mediated both high glucose and palmitate-induced increase in reactive oxygen species levels and decrease in endothelial nitric oxide synthase. Blocking P2X7 inhibited high glucose and palmitate-induced expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 as well as leukocyte-endothelial cell adhesion. Interestingly, high glucose and palmitate enhanced endothelial cell permeability that was dependent on both P2X7 and P2X4. Furthermore, antagonizing the P2X7 inhibited high glucose and palmitate-mediated activation of p38-mitogen

  16. Biodegradation studies of oil sludge containing high hydrocarbons concentration

    Energy Technology Data Exchange (ETDEWEB)

    Olguin-Lora, P.; Munoz-Colunga, A.; Castorena-Cortes, G.; Roldan-Carrillo, T.; Quej Ake, L.; Reyes-Avila, J.; Zapata-Penasco, I.; Marin-Cruz, J.

    2009-07-01

    Oil industry has a significant impact on environment due to the emission of, dust, gases, waste water and solids generated during oil production all the way to basic petrochemical product manufacturing stages. the aim of this work was to evaluate the biodegradation of sludge containing high hydrocarbon concentration originated by a petroleum facility. A sludge sampling was done at the oil residuals pool (ORP) on a gas processing center. (Author)

  17. Particle sedimentation monitoring in high-concentration slurries

    Science.gov (United States)

    Nagasawa, Yoshihiro; Kato, Zenji; Tanaka, Satoshi

    2016-11-01

    In this study, the sedimentation states of particles in high-concentration slurries were elucidated by monitoring their internal states. We prepared transparent high-concentration silica slurries by adjusting the refractive index of the aqueous glycerol liquid in which the particles were dispersed to match that of the silica particles. In addition, a fluorescent dye was dissolved in the liquid. Then, we directly observed the individual and flocculated particles in the slurries during sedimentation by confocal laser scanning fluorescent microscopy. The particles were found to sediment very slowly while exhibiting fluctuating motion. The particle sedimentation rate in the high-concentration slurry with the aqueous glycerol solution (η =0.068 Pa. s ) and a particle volume fraction on the order of 0.3 was determined to be 1.58 ± 0.66 μ m. min-1 on the basis of the obtained image sequences for 24.9 h. In-situ observation provides a large amount of information about the sedimentation behavior of particles in condensed matter.

  18. Improved Dispersion of Carbon Nanotubes in Polymers at High Concentrations

    Science.gov (United States)

    Liu, Chao-Xuan; Choi, Jin-Woo

    2012-01-01

    The polymer nanocomposite used in this work comprises elastomer poly(dimethylsiloxane) (PDMS) as a polymer matrix and multi-walled carbon nanotubes (MWCNTs) as a conductive nanofiller. To achieve uniform distribution of carbon nanotubes within the polymer, an optimized dispersion process was developed, featuring a strong organic solvent—chloroform, which dissolved PDMS base polymer easily and allowed high quality dispersion of MWCNTs. At concentrations as high as 9 wt.%, MWCNTs were dispersed uniformly through the polymer matrix, which presented a major improvement over prior techniques. The dispersion procedure was optimized via extended experimentation, which is discussed in detail. PMID:28348312

  19. Feasibility Study on High Concentrating Photovoltaic Power Towers

    Science.gov (United States)

    Frohberger, Dirk; Jaus, Joachim; Wiesenfarth, Maike; Schramek, Philipp; Bett, Andreas W.

    2010-10-01

    This paper presents an analysis on the concept of high concentrating PV power towers. A feasibility study is conducted in order to evaluate the future potential of this technology. Objective of the analysis is to provide an improved basis for establishing research and development priorities for the PV power tower concept. Performance assessments and cost calculations for a 1 MW prototype PV tower power are derived. Based on the assumption of a highly homogeneously illuminated receiver, levelized costs of electricity of 0.29 €/kWh have been calculated for a prototype PV tower power.

  20. Effects of high nitrogen concentrations on the growth of submersed macrophytes at moderate phosphorus concentrations.

    Science.gov (United States)

    Yu, Qing; Wang, Hong-Zhu; Li, Yan; Shao, Jian-Chun; Liang, Xiao-Min; Jeppesen, Erik; Wang, Hai-Jun

    2015-10-15

    Eutrophication of lakes leading to loss of submersed macrophytes and higher turbidity is a worldwide phenomenon, attributed to excessive loading of phosphorus (P). However, recently, the role of nitrogen (N) for macrophyte recession has received increasing attention. Due to the close relationship between N and P loading, disentanglement of the specific effects of these two nutrients is often difficult, and some controversy still exists as to the effects of N. We studied the effects of N on submersed macrophytes represented by Vallisneria natans (Lour.) Hara in pots positioned at three depths (0.4 m, 0.8 m, and 1.2 m to form a gradient of underwater light conditions) in 10 large ponds having moderate concentrations of P (TP 0.03 ± 0.04 mg L(-1)) and five targeted concentrations of total nitrogen (TN) (0.5, 2, 10, 20, and 100 mg L(-1)), there were two ponds for each treatment. To study the potential shading effects of other primary producers, we also measured the biomass of phytoplankton (ChlaPhyt) and periphyton (ChlaPeri) expressed as chlorophyll a. We found that leaf length, leaf mass, and root length of macrophytes declined with increasing concentrations of TN and ammonium, while shoot number and root mass did not. All the measured growth indices of macrophytes declined significantly with ChlaPhyt, while none were significantly related to ChlaPeri. Neither ChlaPhyt nor ChlaPeri were, however, significantly negatively related to the various N concentrations. Our results indicate that shading by phytoplankton unrelated to the variation in N loading and perhaps toxic stress exerted by high nitrogen were responsible for the decline in macrophyte growth.

  1. Wet oxidation of real coke wastewater containing high thiocyanate concentration.

    Science.gov (United States)

    Oulego, Paula; Collado, Sergio; Garrido, Laura; Laca, Adriana; Rendueles, Manuel; Díaz, Mario

    2014-01-01

    Coke wastewaters, in particular those with high thiocyanate concentrations, represent an important environmental problem because of their very low biodegradability. In this work, the treatment by wet oxidation of real coke wastewaters containing concentrations of thiocyanate above 17 mM has been studied in a 1-L semi-batch reactor at temperatures between 453 and 493 K, with total oxygen pressures in the range of 2.0-8.0 MPa. A positive effect of the matrix of real coke wastewater was observed, resulting in faster thiocyanate degradation than was obtained with synthetic wastewaters. Besides, the effect of oxygen concentration and temperature on thiocyanate wet oxidation was more noticeable in real effluents than in synthetic wastewaters containing only thiocyanate. It was also observed that the degree of mineralization of the matrix organic compounds was higher when the initial thiocyanate concentration increased. Taking into account the experimental data, kinetic models were obtained, and a mechanism implying free radicals was proposed for thiocyanate oxidation in the matrix considered. In all cases, sulphate, carbonates and ammonium were identified as the main reaction products of thiocyanate wet oxidation.

  2. Exposure to High Glucose Concentration Decreases Cell Surface ABCA1 and HDL Biogenesis in Hepatocytes.

    Science.gov (United States)

    Tsujita, Maki; Hossain, Mohammad Anwar; Lu, Rui; Tsuboi, Tomoe; Okumura-Noji, Kuniko; Yokoyama, Shinji

    2017-04-19

    To study atherosclerosis risk in diabetes, we investigated ATP-binding cassette transporter A1 (ABCA1) expression and high-density lipoprotein (HDL) biogenesis in the liver and hepatocytes under hyperglycemic conditions. In streptozotocin-induced diabetic mice, plasma HDL decreased while ABCA1 protein increased without changing its mRNA in the liver, only in the animals that responded to the treatment to show hypoinsulinemia and fasting hyperglycemia but not in the poor responders not showing those. To study the mechanism for this finding, hepatocytes were isolated from the control and diabetic mice, and they showed no difference in expression of ABCA1 protein, its mRNA, and HDL biogenesis in 1 g/l d-glucose but showed decreased HDL biogenesis in 4.5 g/l d-glucose although ABCA1 protein increased without change in its mRNA. Similar findings were confirmed in HepG2 cells with d-glucose but not with l-glucose. Thus, these cell models reproduced the in vivo findings in hyperglycemia. Labeling of cell surface protein revealed that surface ABCA1 decreased in high concentration of d-glucose in HepG2 cells despite the increase of cellular ABCA1 while not with l-glucose. Immunostaining of ABCA1 in HepG2 cells demonstrated the decrease of surface ABCA1 but increase of intracellular ABCA1 with high d-glucose. Clearance of ABCA1 was retarded both in primary hepatocytes and HepG2 cells exposed to high d-glucose but not to l-glucose, being consistent with the decrease of surface ABCA1. It is suggested that localization of ABCA1 to the cell surface is decreased in hepatocytes in hyperglycemic condition to cause decrease of HDL biogenesis.

  3. Cumulative "roof effect" in high-resolution in vivo 31P NMR spectra of human calf muscle and the Clebsch-Gordan coefficients of ATP at 1.5 T.

    Science.gov (United States)

    Schröder, Leif; Schmitz, Christian; Bachert, Peter

    2005-05-01

    NMR spectra of non-weakly coupled spin systems exhibit asymmetries in line intensities known as "roof effect" in 1D spectroscopy. Due to limited spectral resolution, this effect has not been paid much attention so far in in vivo spectroscopy. But when high-quality spectra are obtained, this effect should be taken into account to explain the quantum-mechanical fine structure of the system. Adenosine 5'-triphosphate (ATP) represents a 31P spin system with multiple line splittings which are caused by J-couplings of medium strength at 1.5 T. We analyzed the ATP roof effect in vivo, especially for the beta-ATP multiplet. The intensities of its outer resonances deviate by ca. 12.5% from a symmetrical triplet. As this asymmetry reflects the transition from Paschen-Back to Zeeman effect with total spin that is largely broken up, the Clebsch-Gordan coefficients of the system can be indicated in analogy to the hyperfine structure of hydrogen. Taking the roof effect into account, the chi2 of fitting in vivo ATP resonances is reduced by ca. 9% (p<0.005).

  4. Cumulative “roof effect” in high-resolution in vivo 31P NMR spectra of human calf muscle and the Clebsch Gordan coefficients of ATP at 1.5 T

    Science.gov (United States)

    Schröder, Leif; Schmitz, Christian; Bachert, Peter

    2005-05-01

    NMR spectra of non-weakly coupled spin systems exhibit asymmetries in line intensities known as "roof effect" in 1D spectroscopy. Due to limited spectral resolution, this effect has not been paid much attention so far in in vivo spectroscopy. But when high-quality spectra are obtained, this effect should be taken into account to explain the quantum-mechanical fine structure of the system. Adenosine 5'-triphosphate (ATP) represents a 31P spin system with multiple line splittings which are caused by J-couplings of medium strength at 1.5 T. We analyzed the ATP roof effect in vivo, especially for the β-ATP multiplet. The intensities of its outer resonances deviate by ca. 12.5% from a symmetrical triplet. As this asymmetry reflects the transition from Paschen-Back to Zeeman effect with total spin that is largely broken up, the Clebsch-Gordan coefficients of the system can be indicated in analogy to the hyperfine structure of hydrogen. Taking the roof effect into account, the χ2 of fitting in vivo ATP resonances is reduced by ca. 9% ( p < 0.005).

  5. Action of Al-ATP on the isolated working rat heart.

    Science.gov (United States)

    Korchazhkina, O; Wright, G; Exley, C

    1998-02-15

    ATP is an important extracellular messenger in the coronary vasculature of the heart. To be effective its extracellular concentration must be tightly controlled and this is achieved via ectonucleotidases located in the luminal surface of the coronary endothelium. Al-ATP is a potent inhibitor of the hydrolysis of ATP and we speculated that Al-ATP released by cells into the blood would disrupt the signalling function of extracellular ATP. We tested this hypothesis by perfusing isolated working Wistar rat hearts with buffers containing either ATP or Al-ATP. The functional parameters measured were, coronary flow, heart rate and pulsatile power. A number of control perfusions including adenosine, ATP-gamma-S and Al were used to identify those effects which might be specific to ATP and Al-ATP. Al-ATP did not appear to inhibit the function of the endothelial ectonucleotidases. Both ATP and Al-ATP produced a significant increase in coronary flow and this could be attributed to a coronary vasodilation. Interestingly, whilst the effect of ATP was reversible that of Al-ATP was not. ATP caused a reduction in heart rate which was potentiated by aluminium. The negatively chronotropic effect of Al-ATP was mediated via a mechanism which was either distinct from or in addition to the similar response known to be caused by adenosine. We have demonstrated for the first time an influence of Al-ATP on heart function. Perhaps more pertinently we present the first evidence that Al-ATP may influence the function of ATP-specific receptors.

  6. Electrochemical sensing of ATP with synthetic cyclophane as recognition element

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    A new electrochemical sensor for ATP with synthetic cyclophane stably attached onto single-walled carbon nanotubes (SWNTs) as the recognition elements is described. UV-vis and cyclic voltammetric results demonstrate that ATP may interact with the synthetic cyclophane recognition elements to form a stable adduct mainly through electrostatic, π-π stacking and donor-acceptor interactions. Such interactions eventually lead to a decrease in the peak currents of the cyclophane recognition elements attached onto the SWNT electronic transducer, which could be used for electrochemical sensing of ATP. Under the conditions employed here, the ratio of the decrease in the anodic peak current is linear with ATP concentration within a concentration range from 10 to 120 μM with a linear coefficiency of 0.993. This study may offer a new and simple electrochemical approach for effective sensing of ATP.

  7. Strategies for the production of high concentrations of bioethanol from seaweeds: production of high concentrations of bioethanol from seaweeds.

    Science.gov (United States)

    Yanagisawa, Mitsunori; Kawai, Shigeyuki; Murata, Kousaku

    2013-01-01

    Bioethanol has attracted attention as an alternative to petroleum-derived fuel. Seaweeds have been proposed as some of the most promising raw materials for bioethanol production because they have several advantages over lignocellulosic biomass. However, because seaweeds contain low contents of glucans, i.e., polysaccharides composed of glucose, the conversion of only the glucans from seaweed is not sufficient to produce high concentrations of ethanol. Therefore, it is also necessary to produce ethanol from other specific carbohydrate components of seaweeds, including sulfated polysaccharides, mannitol, alginate, agar and carrageenan. This review summarizes the current state of research on the production of ethanol from seaweed carbohydrates for which the conversion of carbohydrates to sugars is a key step and makes comparisons with the production of ethanol from lignocellulosic biomass. This review provides valuable information necessary for the production of high concentrations of ethanol from seaweeds.

  8. Model study of ATP and ADP buffering, transport of Ca(2+) and Mg(2+), and regulation of ion pumps in ventricular myocyte

    Science.gov (United States)

    Michailova, A.; McCulloch, A.

    2001-01-01

    We extended the model of the ventricular myocyte by Winslow et al. (Circ. Res 1999, 84:571-586) by incorporating equations for Ca(2+) and Mg(2+) buffering and transport by ATP and ADP and equations for MgATP regulation of ion transporters (Na(+)-K(+) pump, sarcolemmal and sarcoplasmic Ca(2+) pumps). The results indicate that, under normal conditions, Ca(2+) binding by low-affinity ATP and diffusion of CaATP may affect the amplitude and time course of intracellular Ca(2+) signals. The model also suggests that a fall in ATP/ADP ratio significantly reduces sarcoplasmic Ca(2+) content, increases diastolic Ca(2+), lowers systolic Ca(2+), increases Ca(2+) influx through L-type channels, and decreases the efficiency of the Na(+)/Ca(2+) exchanger in extruding Ca(2+) during periodic voltage-clamp stimulation. The analysis suggests that the most important reason for these changes during metabolic inhibition is the down-regulation of the sarcoplasmic Ca(2+)-ATPase pump by reduced diastolic MgATP levels. High Ca(2+) concentrations developed near the membrane might have a greater influence on Mg(2+), ATP, and ADP concentrations than that of the lower Ca(2+) concentrations in the bulk myoplasm. The model predictions are in general agreement with experimental observations measured under normal and pathological conditions.

  9. The E646D-ATP13A4 mutation associated with autism reveals a defect in calcium regulation.

    Science.gov (United States)

    Vallipuram, Janaki; Grenville, Jeffrey; Crawford, Dorota A

    2010-03-01

    ATP13A4 is a member of the subfamily of P5-type ATPases. P5-type ATPases are the least studied of the P-type ATPase subfamilies with no ion specificities assigned to them. In order to elucidate ATP13A4 function, we studied the protein's subcellular localization and tested whether it is involved in calcium regulation. The intracellular calcium concentration was measured in COS-7 cells over-expressing mouse ATP13A4 using ratiometric calcium imaging with fura-2 AM as a calcium indicator. The results of this study show that ATP13A4 is localized to the endoplasmic reticulum (ER). Furthermore, we demonstrate that over-expression of ATP13A4 in COS-7 cells caused a significant increase in the intracellular calcium level. Interestingly, over-expression of the sequence variant containing a substitution of aspartic acid for a glutamic acid (E646D), previously found in patients with autism spectrum disorder (ASD), did not increase the free cellular calcium likely due to the mutation. In this study, we also describe the expression of ATP13A4 during mouse embryonic development. Quantitative real-time PCR revealed that ATP13A4 was highly expressed at embryonic days 15-17, when neurogenesis takes place. The present study is the first to provide further insights into the biological role of a P5-type ATPase. Our results demonstrate that ATP13A4 may be involved in calcium regulation and that its expression is developmentally regulated. Overall, this study provides support for the hypothesis that ATP13A4 may play a vital role in the developing nervous system and its impairment can contribute to the symptoms seen in ASD.

  10. The P2X7 loss-of-function Glu496Ala polymorphism affects ex vivo cytokine release and protects against the cytotoxic effects of high ATP-levels

    Directory of Open Access Journals (Sweden)

    Wesselius Anke

    2012-12-01

    Full Text Available Abstract Background The P2X7 receptor plays an important role in cytokine release during the inflammatory response in vivo. Polymorphisms within the P2X7 receptor gene that lead to loss of receptor function may contribute to impaired cytokine release by immune cells. Therefore, we investigated whether a known loss-of-function polymorphism (Glu496Ala in the P2X7 receptor gene leads to alterations in cytokine release in response to ATP. Results An ex vivo whole blood model was used to induce an inflammatory reaction with the pro-inflammatory stimuli LPS and PHA (phytohemagglutinin. Blood from n=9 subjects with the Glu496Ala P2X7 SNP (P2X7MUT and n=7 ‘wild-type’ subjects (no P2X7 SNP; P2X7WT was used. Addition of ATP (0.9-3 mM to LPS/PHA-stimulated whole blood induced an increase in IL-1β release in P2X7MUT subjects, whereas decreased release was observed in P2X7WT subjects. Decreased levels of IL-6 and TNF-α in response to ATP were shown in both P2X7MUT and P2X7WT subjects, which was less pronounced in P2X7MUT subjects. ATP at 3 mM also significantly decreased levels of lactate dehydrogenase (LDH in P2X7MUT subjects compared to P2X7WT subjects. Conclusions The presence of the non-synonymous Glu496Ala loss-of-function polymorphism within the P2X7 receptor gene is likely to be of importance in the release of cytokines during inflammation. Furthermore, this study suggests that carriers of the Glu496Ala loss-of-function polymorphism are protected against the cytotoxic effects of high ATP-levels.

  11. MicroRNA-144 regulates hepatic ATP binding cassette transporter A1 and plasma high-density lipoprotein after activation of the nuclear receptor farnesoid X receptor.

    Science.gov (United States)

    de Aguiar Vallim, Thomas Q; Tarling, Elizabeth J; Kim, Tammy; Civelek, Mete; Baldán, Ángel; Esau, Christine; Edwards, Peter A

    2013-06-07

    The bile acid receptor farnesoid X receptor (FXR) regulates many aspects of lipid metabolism by variouscomplex and incompletely understood molecular mechanisms. We set out to investigate the molecular mechanisms for FXR-dependent regulation of lipid and lipoprotein metabolism. To identify FXR-regulated microRNAs that were subsequently involved in regulating lipid metabolism. ATP binding cassette transporter A1 (ABCA1) is a major determinant of plasma high-density lipoprotein (HDL)-cholesterol levels. Here, we show that activation of the nuclear receptor FXR in vivo increases hepatic levels of miR-144, which in turn lowers hepatic ABCA1 and plasma HDL levels. We identified 2 complementary sequences to miR-144 in the 3' untranslated region of ABCA1 mRNA that are necessary for miR-144-dependent regulation. Overexpression of miR-144 in vitro decreased both cellular ABCA1 protein and cholesterol efflux to lipid-poor apolipoprotein A-I protein, whereas overexpression in vivo reduced hepatic ABCA1 protein and plasma HDL-cholesterol. Conversely, silencing miR-144 in mice increased hepatic ABCA1 protein and HDL-cholesterol. In addition, we used tissue-specific FXR-deficient mice to show that induction of miR-144 and FXR-dependent hypolipidemia requires hepatic, but not intestinal, FXR. Finally, we identified functional FXR response elements upstream of the miR-144 locus, consistent with direct FXR regulation. We have identified a novel pathway involving FXR, miR-144, and ABCA1 that together regulate plasma HDL-cholesterol.

  12. Calcium and ATP control multiple vital functions

    Science.gov (United States)

    Verkhratsky, Alexei

    2016-01-01

    Life on Planet Earth, as we know it, revolves around adenosine triphosphate (ATP) as a universal energy storing molecule. The metabolism of ATP requires a low cytosolic Ca2+ concentration, and hence tethers these two molecules together. The exceedingly low cytosolic Ca2+ concentration (which in all life forms is kept around 50–100 nM) forms the basis for a universal intracellular signalling system in which Ca2+ acts as a second messenger. Maintenance of transmembrane Ca2+ gradients, in turn, requires ATP-dependent Ca2+ transport, thus further emphasizing the inseparable links between these two substances. Ca2+ signalling controls the most fundamental processes in the living organism, from heartbeat and neurotransmission to cell energetics and secretion. The versatility and plasticity of Ca2+ signalling relies on cell specific Ca2+ signalling toolkits, remodelling of which underlies adaptive cellular responses. Alterations of these Ca2+ signalling toolkits lead to aberrant Ca2+ signalling which is fundamental for the pathophysiology of numerous diseases from acute pancreatitis to neurodegeneration. This paper introduces a theme issue on this topic, which arose from a Royal Society Theo Murphy scientific meeting held in March 2016. This article is part of the themed issue ‘Evolution brings Ca2+ and ATP together to control life and death’. PMID:27377728

  13. Chronic Diarrhea Associated with High Teriflunomide Blood Concentration

    OpenAIRE

    Duquette, André; Frenette, Anne Julie; Doré, Maxime

    2016-01-01

    Objective To report the case of a patient treated with leflunomide that presented with chronic diarrhea associated with high teriflunomide blood concentration. Case Summary An 84-year-old woman taking leflunomide 20 mg once daily for the past 2 years to treat rheumatoid arthritis (RA) was investigated for severe chronic diarrhea that had been worsening for the past 5 months. The patient’s general condition progressively deteriorated and included electrolyte imbalances and a transient loss of ...

  14. Effect of high soil copper concentration on mycorrhizal grapevines

    Science.gov (United States)

    Nogales, Amaia; Santos, Erika S.; Viegas, Wanda; Aran, Diego; Pereira, Sofia H.; Vidigal, Patricia; Lopes, Carlos M.; Abreu, M. Manuela

    2017-04-01

    Repeated application of Copper (Cu) based fungicides in vineyards since the end of the 19th century has led to a significant increase in the concentration of this chemical element in many viticultural soils. Although Cu is an essential micronutrient for most organisms, it can be toxic for the development and survival of plants and soil (micro)organisms at high concentrations and eventually lead to yield loses in viticulture, as it negatively affects key physiological and biogeochemical processes. However, some soil microorganisms, including arbuscular mycorrhizal fungi (AMF), have developed adaptive mechanisms for persistence in environments with supra-optimal levels of essential elements or in the presence of harmful ones, as well as for increasing plant tolerance to such abiotic stress conditions. The objective of this work was to evaluate the effect of a high total soil concentration of Cu on microbial soil activity as well as on the development of mycorrhizal and non-mycorrhizal grapevines. A microcosm assay was set up under greenhouse and controlled conditions. Touriga Nacional grapevine variety plants grafted onto 1103P rootstocks were inoculated either with the AMF Rhizophagus irregularis or Funneliformis mosseae, or were left as non-inoculated controls. After three months, they were transplanted to containers filled with 4 kg of a sandy soil (pH: 7.0; electrical conductivity: 0.08 mS/cm; [organic C]: 5.6 g/kg; [N-NO3]: 1.1 mg/kg; [N-NH4]: 2.5 mg/kg; [extractable K]: 45.1 mg/kg; [extractable P]: 52.3 mg/kg), collected near to a vineyard in Pegões (Portugal). Two treatments were carried out: with and without Cu application. The soil with high Cu concentration was prepared by adding 300 mg Cu/kg (in the form of an aqueous solution of CuSO4·5H2O) followed by an incubation during four weeks in plastic bags at room temperature in dark. Physico-chemical soil characteristics (pH, electrical conductivity and nutrients concentration in available fraction), soil

  15. F1-dependent translation of mitochondrially encoded Atp6p and Atp8p subunits of yeast ATP synthase

    OpenAIRE

    Rak, Malgorzata; Tzagoloff, Alexander

    2009-01-01

    The ATP synthase of yeast mitochondria is composed of 17 different subunit polypeptides. We have screened a panel of ATP synthase mutants for impaired expression of Atp6p, Atp8p, and Atp9p, the only mitochondrially encoded subunits of ATP synthase. Our results show that translation of Atp6p and Atp8p is activated by F1 ATPase (or assembly intermediates thereof). Mutants lacking the α or β subunits of F1, or the Atp11p and Atp12p chaperones that promote F1 assembly, have normal levels of the b...

  16. Particulate Matter Concentrations in East Oakland's High Street Corridor

    Science.gov (United States)

    Lei, P.; Jackson, J.; Lewis, R.; Marigny, A.; Mitchell, J. D.; Nguyen, R.; Philips, B.; Randle, D.; Romero, D.; Spears, D.; Telles, C.; Weissman, D.

    2012-12-01

    Particulate matter (PM) is a complex mixture of small solid pieces and/or liquid droplets in the air. High concentrations of PM can pose a serious health hazard because inhalation can result in breathing problems and/or aggravate asthma. Long term exposure can increase the likelihood of respiratory problems like asthma and emphysema as well as cancer. The smaller the particles, the deeper they can get into the respiratory system. For this reason, the smallest particles, those smaller than 2.5 micrometers in diameter (PM2.5), are the most dangerous. PM2.5 is largely emitted from motor vehicles burning fuels that don't break down fully. Our research team investigated the levels of PM2.5 as well as particles smaller than 10 micrometers (PM10) and total suspended particulate (TSP) along the northeast-southwest trending High Street Corridor, near Fremont High School in East Oakland, California. Using the Aerocet 531 mass particle counter, team members walked through neighborhoods and along major roads within a 1 mile radius of Fremont High School. The Aerocet 531 recorded two minute average measurements of all the relevant PM sizes, which are reported in mg/m3. Measurements were consistently taken in the morning, between 8:30 and 11:30 am. Preliminary results indicate maximum readings of all PM sizes at sites that are in close proximity to a major freeway (Interstate-880). These results support our initial hypothesis that proximity to major roads and freeways, especially those with high diesel-fuel burning truck traffic, would be the primary factor affecting PM concentration levels. Preliminary median and maximum readings all suggest particulate matter levels below what the EPA would consider unhealthy or risky.

  17. High manganese concentrations in rocks at Gale crater, Mars

    Science.gov (United States)

    Lanza, Nina L.; Fischer, Woodward W.; Wiens, Roger C.; Grotzinger, John; Ollila, Ann M.; Anderson, Ryan B.; Clark, Benton C.; Gellert, Ralf; Mangold, Nicolas; Maurice, Sylvestre; Le Mouélic, Stéphane; Nachon, Marion; Schmidt, Mariek E.; Berger, Jeffrey; Clegg, Samuel M.; Forni, Olivier; Hardgrove, Craig; Melikechi, Noureddine; Newsom, Horton E.; Sautter, Violaine

    2014-01-01

    The surface of Mars has long been considered a relatively oxidizing environment, an idea supported by the abundance of ferric iron phases observed there. However, compared to iron, manganese is sensitive only to high redox potential oxidants, and when concentrated in rocks, it provides a more specific redox indicator of aqueous environments. Observations from the ChemCam instrument on the Curiosity rover indicate abundances of manganese in and on some rock targets that are 1–2 orders of magnitude higher than previously observed on Mars, suggesting the presence of an as-yet unidentified manganese-rich phase. These results show that the Martian surface has at some point in time hosted much more highly oxidizing conditions than has previously been recognized.

  18. Shock initiation studies on high concentration hydrogen peroxide

    Energy Technology Data Exchange (ETDEWEB)

    Sheffield, Stephen A [Los Alamos National Laboratory; Dattelbaum, Dana M [Los Alamos National Laboratory; Stahl, David B [Los Alamos National Laboratory; Gibson, L. Lee [Los Alamos National Laboratory; Bartram, Brian D. [Los Alamos National Laboratory

    2009-01-01

    Concentrated hydrogen peroxide (H{sub 2}O{sub 2}) has been known to detonate for many years. However, because of its reactivity and the difficulty in handling and confining it, along with the large critical diameter, few studies providing basic information about the initiation and detonation properties have been published. We are conducting a study to understand and quantify the initiation and detonation properties of highly concentrated H{sub 2}O{sub 2} using a gas-driven two-stage gun to produce well defined shock inputs. Multiple magnetic gauges are used to make in-situ measurements of the growth of reaction and subsequent detonation in the liquid. These experiments are designed to be one-dimensional to eliminate any difficulties that might be encountered with large critical diameters. Because of the concern of the reactivity of the H{sub 2}O{sub 2} with the confining materials, a remote loading system has been developed. The gun is pressurized, then the cell is filled and the experiment shot within less than three minutes. TV cameras are attached to the target so the cell filling can be monitored. Several experiments have been completed on {approx}98 wt % H{sub 2}O{sub 2}/H{sub 2}O mixtures; initiation has been observed in some experiments that shows homogeneous shock initiation behavior. The initial shock pressurizes and heats the mixture. After an induction time, a thermal explosion type reaction produces an evolving reactive wave that strengthens and eventually overdrives the first wave producing a detonation. From these measurements, we have determined unreacted Hugoniot information, times (distances) to detonation (Pop-plot points) that indicate low sensitivity, and detonation velocities of high concentration H{sub 2}O{sub 2}/H{sub 2}O solutions that agree with earlier estimates.

  19. Shock initiation studies on high concentration hydrogen peroxide

    Energy Technology Data Exchange (ETDEWEB)

    Sheffield, Stephen A [Los Alamos National Laboratory; Dattelbaum, Dana M [Los Alamos National Laboratory; Stahl, David B [Los Alamos National Laboratory; Gibson, L. Lee [Los Alamos National Laboratory; Bartram, Brian D. [Los Alamos National Laboratory

    2009-01-01

    Concentrated hydrogen peroxide (H{sub 2}O{sub 2}) has been known to detonate for many years. However, because of its reactivity and the difficulty in handling and confining it, along with the large critical diameter, few studies providing basic information about the initiation and detonation properties have been published. We are conducting a study to understand and quantify the initiation and detonation properties of highly concentrated H{sub 2}O{sub 2} using a gas-driven two-stage gun to produce well defined shock inputs. Multiple magnetic gauges are used to make in-situ measurements of the growth of reaction and subsequent detonation in the liquid. These experiments are designed to be one-dimensional to eliminate any difficulties that might be encountered with large critical diameters. Because of the concern of the reactivity of the H{sub 2}O{sub 2} with the confining materials, a remote loading system has been developed. The gun is pressurized, then the cell is filled and the experiment shot within less than three minutes. TV cameras are attached to the target so the cell filling can be monitored. Several experiments have been completed on {approx}98 wt % H{sub 2}O{sub 2}/H{sub 2}O mixtures; initiation has been observed in some experiments that shows homogeneous shock initiation behavior. The initial shock pressurizes and heats the mixture. After an induction time, a thermal explosion type reaction produces an evolving reactive wave that strengthens and eventually overdrives the first wave producing a detonation. From these measurements, we have determined unreacted Hugoniot information, times (distances) to detonation (Pop-plot points) that indicate low sensitivity, and detonation velocities of high concentration H{sub 2}O{sub 2}/H{sub 2}O solutions that agree with earlier estimates.

  20. Functional expression of a heterologous nickel-dependent, ATP-independent urease in Saccharomyces cerevisiae.

    Science.gov (United States)

    Milne, N; Luttik, M A H; Cueto Rojas, H F; Wahl, A; van Maris, A J A; Pronk, J T; Daran, J M

    2015-07-01

    In microbial processes for production of proteins, biomass and nitrogen-containing commodity chemicals, ATP requirements for nitrogen assimilation affect product yields on the energy producing substrate. In Saccharomyces cerevisiae, a current host for heterologous protein production and potential platform for production of nitrogen-containing chemicals, uptake and assimilation of ammonium requires 1 ATP per incorporated NH3. Urea assimilation by this yeast is more energy efficient but still requires 0.5 ATP per NH3 produced. To decrease ATP costs for nitrogen assimilation, the S. cerevisiae gene encoding ATP-dependent urease (DUR1,2) was replaced by a Schizosaccharomyces pombe gene encoding ATP-independent urease (ure2), along with its accessory genes ureD, ureF and ureG. Since S. pombe ure2 is a Ni(2+)-dependent enzyme and Saccharomyces cerevisiae does not express native Ni(2+)-dependent enzymes, the S. pombe high-affinity nickel-transporter gene (nic1) was also expressed. Expression of the S. pombe genes into dur1,2Δ S. cerevisiae yielded an in vitro ATP-independent urease activity of 0.44±0.01 µmol min(-1) mg protein(-1) and restored growth on urea as sole nitrogen source. Functional expression of the Nic1 transporter was essential for growth on urea at low Ni(2+) concentrations. The maximum specific growth rates of the engineered strain on urea and ammonium were lower than those of a DUR1,2 reference strain. In glucose-limited chemostat cultures with urea as nitrogen source, the engineered strain exhibited an increased release of ammonia and reduced nitrogen content of the biomass. Our results indicate a new strategy for improving yeast-based production of nitrogen-containing chemicals and demonstrate that Ni(2+)-dependent enzymes can be functionally expressed in S. cerevisiae.

  1. Source identification of high glyme concentrations in the Oder River.

    Science.gov (United States)

    Stepien, D K; Püttmann, W

    2014-05-01

    The objective of the following study was to identify the source of high concentrations of glycol diethers (diglyme, triglyme, and tetraglyme) in the Oder River. Altogether four sampling campaigns were conducted and over 50 surface samples collected. During the first two samplings of the Oder River in the Oderbruch region (km 626-690), glymes were detected at concentrations reaching 0.065 μg L(-1) (diglyme), 0.54 μg L(-1) (triglyme) and 1.7 μg L(-1) (tetraglyme). The subsequent sampling of the Oder River, from the area close to the source to the Poland-Germany border (about 500 km) helped to identify the possible area of the dominating glyme entry into the river between km 310 and km 331. During that sampling, the maximum concentration of triglyme was 0.46 μg L(-1) and tetraglyme 2.2 μg L(-1); diglyme was not detected. The final sampling focused on the previously identified area of glyme entry, as well as on tributaries of the Oder River. Samples from Czarna Woda stream and Kaczawa River contained even higher concentrations of diglyme, triglyme, and tetraglyme, reaching 5.2 μg L(-1), 13 μg L(-1) and 81 μg L(-1), respectively. Finally, three water samples were analyzed from a wastewater treatment plant receiving influents from a Copper Smelter and Refinery; diglyme, triglyme, and tetraglyme were present at a maximum concentration of 1700 μg L(-1), 13,000 μg L(-1), and 190,000 μg L(-1), respectively. Further research helped to identify the source of glymes in the wastewater. The gas desulfurization process Solinox uses a mixture of glymes (Genosorb(®)1900) as a physical absorption medium to remove sulfur dioxide from off-gases from the power plant. The wastewater generated from the process and from the maintenance of the equipment is initially directed to the wastewater treatment plant where it undergoes mechanical and chemical treatment processes before being discharged to the tributaries of the Oder River. Although monoglyme was

  2. F1 rotary motor of ATP synthase is driven by the torsionally-asymmetric drive shaft

    Science.gov (United States)

    Kulish, O.; Wright, A. D.; Terentjev, E. M.

    2016-06-01

    F1F0 ATP synthase (ATPase) either facilitates the synthesis of ATP in a process driven by the proton moving force (pmf), or uses the energy from ATP hydrolysis to pump protons against the concentration gradient across the membrane. ATPase is composed of two rotary motors, F0 and F1, which compete for control of their shared γ -shaft. We present a self-consistent physical model of F1 motor as a simplified two-state Brownian ratchet using the asymmetry of torsional elastic energy of the coiled-coil γ -shaft. This stochastic model unifies the physical concepts of linear and rotary motors, and explains the stepped unidirectional rotary motion. Substituting the model parameters, all independently known from recent experiments, our model quantitatively reproduces the ATPase operation, e.g. the ‘no-load’ angular velocity is ca. 400 rad/s anticlockwise at 4 mM ATP. Increasing the pmf torque exerted by F0 can slow, stop and overcome the torque generated by F1, switching from ATP hydrolysis to synthesis at a very low value of ‘stall torque’. We discuss the motor efficiency, which is very low if calculated from the useful mechanical work it produces - but is quite high when the ‘useful outcome’ is measured in the number of H+ pushed against the chemical gradient.

  3. [Stabilization of Cadmium Contaminated Soils by Ferric Ion Modified Attapulgite (Fe/ATP)--Characterizations and Stabilization Mechanism].

    Science.gov (United States)

    Rong, Yang; Li, Rong-bo; Zhou, Yong-li; Chen, Jing; Wang, Lin-ling; Lu, Xiao-hua

    2015-08-01

    Ferric ion modified attapulgite (Fe/ATP) was prepared by impregnation and its structure and morphology were characterized. The toxicity characteristic leaching procedure (TCLP) was used to evaluate the effect of Cadmium( Cd) stabilization in soil with the addition of attapulgite (ATP) and Fe/ATP. The stabilization mechanism of Cd was further elucidated by comparing the morphologies and structure of ATP and Fe/ATP before and after Cd adsorption. Fe/ATP exhibited much better adsorption capacity than ATP, suggesting different adsorption mechanisms occurred between ATP and Fe/ATP. The leaching concentrations of Cd in soil decreased by 45% and 91% respectively, with the addition of wt. 20% ATP and Fe/ATP. The former was attributed to the interaction between Cd2 and --OH groups by chemical binding to form inner-sphere complexes in ATP and the attachment between Cd2+ and the defect sites in ATP framework. Whereas Cd stabilization with Fe/ATP was resulted from the fact that the active centers (--OH bonds or O- sites) on ATP could react with Fe3+ giving Fe--O--Cd-- bridges, which helped stabilize Cd in surface soil. What'more, the ferric oxides and metal hydroxides on the surface of ATP could interact with Cd, probably by the formation of cadmium ferrite. In conclusion, Fe/ATP, which can be easily prepared, holds promise as a potential low-cost and environmental friendly stabilizing agent for remediation of soil contaminated with heavy metals.

  4. Catalytic wet air oxidation of high concentration pharmaceutical wastewater.

    Science.gov (United States)

    Zhan, Wei; Wang, Xiaocong; Li, Daosheng; Ren, Yongzheng; Liu, Dongqi; Kang, Jianxiong

    2013-01-01

    In this study, we investigated the pretreatment of a high concentration pharmaceutical wastewater by catalytic wet air oxidation (CWAO) process. Different experiments were conducted to investigate the effects of the catalyst type, operating temperature, initial system pH, and oxygen partial pressure on the oxidation of the wastewater. Results show that the catalysts prepared by the co-precipitation method have better catalytic activity compared to others. Chemical oxygen demand (COD) conversion increased with the increase in temperature from 160 to 220 °C and decreased with the increase in pH. Moreover, the effect of the oxygen partial pressure on the COD conversion was significant only during the first 20 min of the reaction. Furthermore, the biodegradability of the wastewater improved greatly after CWAO, the ratio of BOD5/COD increased less than 0.1-0.75 when treated at 220 °C (BOD: biochemical oxygen demand).

  5. Acquisition and Analysis of Data from High Concentration Solutions

    KAUST Repository

    Besong, Tabot M.D.

    2016-05-13

    The problems associated with ultracentrifugal analysis of macromolecular solutions at high (>10 mg/ml) are reviewed. Especially for the case of solutes which are non-monodisperse, meaningful results are not readily achievable using sedimentation velocity approaches. It is shown however by both simulation and analysis of practical data that using a modified form of an algorithm (INVEQ) published in other contexts, sedimentation equilibrium (SE) profiles can be analysed successfully, enabling topics such as oligomer presence or formation to be defined.To achieve this, it is necessary to employ an approach in which the solution density, which in an SE profile is radius-dependent, is taken into consideration. Simulation suggests that any reasonable level of solute concentration can be analysed.

  6. Rheological properties of highly concentrated protein-stabilized emulsions.

    Science.gov (United States)

    Dimitrova, Tatiana D; Leal-Calderon, Fernando

    2004-05-20

    We prepared concentrated quasi monodisperse hexadecane-in-water emulsions stabilized by various proteins and investigated their rheological properties. Some protein-stabilized emulsions possess remarkably high elasticity and at the same time they are considerably fragile--they exhibit coalescence at yield strain and practically do not flow. The elastic storage modulus G' and the loss modulus G" of the emulsions were determined for different oil volume fractions above the random close packing. Surprisingly, the dimensionless elastic moduli G'/(sigma/a), sigma being the interfacial tension, and a being the mean drop radius, obtained for emulsions stabilized by different proteins do not collapse on a single master curve. They are almost always substantially higher than the corresponding values obtained for equivalent Sodium Dodecyl Sulfate (SDS)-stabilized emulsions. The unusually high elasticity cannot be attributed to a specificity of the continuous phase, because the osmotic equation of state of our emulsions is found identical to the one obtained for samples stabilized by classical surfactants. In parallel, we mimicked the thin films that separate the droplets in the concentrated emulsion and found that the protein adsorption layers contain a substantial number of sticky surface aggregates. These severely obstruct local rearrangements of individual drops in respect to their neighbors which leads to coalescence at yield strain. Furthermore, we found that G'/(sigma/a) is correlated (for a given oil volume fraction) to the dilatational elastic modulus, of the protein layer adsorbed on the droplets. The intrinsic elasticity of the protein layers, together with the blocked local rearrangements are considered as the main factors determining the unusual bulk elasticity of the studied emulsions.

  7. Do high concentrations of microcystin prevent Daphnia control of phytoplankton?

    Science.gov (United States)

    Chislock, Michael F; Sarnelle, Orlando; Jernigan, Lauren M; Wilson, Alan E

    2013-04-15

    Toxin-producing cyanobacteria have frequently been hypothesized to limit the ability of herbivorous zooplankton (such as Daphnia) to control phytoplankton biomass by inhibiting feeding, and in extreme cases, causing zooplankton mortality. Using limnocorral experiments in hyper-eutrophic ponds located in Alabama and Michigan (U.S.A.), we tested the hypothesis that high levels of cyanobacteria and microcystin, a class of hepatotoxins produced by several cyanobacterial genera, prevent Daphnia from strongly reducing phytoplankton abundance. At the start of the first experiment (Michigan), phytoplankton communities were dominated by toxic Microcystis and Anabaena (∼96% of total phytoplankton biomass), and concentrations of microcystin were ∼3 μg L⁻¹. Two weeks after adding Daphnia pulicaria from a nearby eutrophic lake, microcystin levels increased to ∼6.5 μg L⁻¹, yet Daphnia populations increased exponentially (r = 0.24 day⁻¹). By the third week, Daphnia had suppressed phytoplankton biomass by ∼74% relative to the no Daphnia controls and maintained reduced phytoplankton biomass until the conclusion of the five-week experiment. In the second experiment (Alabama), microcystin concentrations were greater than 100 μg L⁻¹, yet a mixture of three D. pulicaria clones from eutrophic lakes in southern MI increased and again reduced phytoplankton biomass, in this case by over 80%. The ability of Daphnia to increase in abundance and suppress phytoplankton biomass, despite high initial levels of cyanobacteria and microcystin, indicates that the latter does not prevent strong control of phytoplankton biomass by Daphnia genotypes that are adapted to environments with abundant cyanobacteria and associated cyanotoxins.

  8. Highly Concentrated Acetic Acid Poisoning: 400 Cases Reviewed

    Directory of Open Access Journals (Sweden)

    Konstantin Brusin

    2012-12-01

    Full Text Available Background: Caustic substance ingestion is known for causing a wide array of gastrointestinal and systemic complications. In Russia, ingestion of acetic acid is a major problem which annually affects 11.2 per 100,000 individuals. The objective of this study was to report and analyze main complications and outcomes of patients with 70% concentrated acetic acid poisoning. Methods: This was a retrospective study of patients with acetic acid ingestion who were treated at Sverdlovsk Regional Poisoning Treatment Center during 2006 to 2012. GI mucosal injury of each patient was assessed with endoscopy according to Zargar’s scale. Data analysis was performed to analyze the predictors of stricture formation and mortality. Results: A total of 400 patients with median age of 47 yr were included. GI injury grade I was found in 66 cases (16.5%, IIa in 117 (29.3%, IIb in 120 (30%, IIIa in 27 (16.7% and IIIb in 70 (17.5%. 11% of patients developed strictures and overall mortality rate was 21%. Main complications were hemolysis (55%, renal injury (35%, pneumonia (27% and bleeding during the first 3 days (27%. Predictors of mortality were age 60 to 79 years, grade IIIa and IIIb of GI injury, pneumonia, stages “I”, “F” and “L” of kidney damage according to the RIFLE scale and administration of prednisolone. Predictors of stricture formation were ingestion of over 100 mL of acetic acid and grade IIb and IIIa of GI injury. Conclusion: Highly concentrated acetic acid is still frequently ingested in Russia with a high mortality rate. Patients with higher grades of GI injury, pneumonia, renal injury and higher amount of acid ingested should be more carefully monitored as they are more susceptible to develop fatal consequences.          

  9. Electrophysiological effects of ATP on brain neurones.

    Science.gov (United States)

    Illes, P; Nieber, K; Nörenberg, W

    1996-12-01

    1. The electrophysiological effects of ATP on brain neurones are either due to the direct activation of P2 purinoceptors by the unmetabolized nucleotide or to the indirect activation of P1. purinoceptors by the degradation product adenosine. 2. Two subtypes of P2 purinoceptors are involved, a ligand-activated ion channel (P2X) and a G protein-coupled receptor (P2Y). Hence, the stimulation of P2X purinoceptors leads to a cationic conductance increase, while the stimulation of P2Y purinoceptors leads to a G protein-mediated opening or closure of potassium channels. 3. ATP may induce a calcium-dependent potassium current by increasing the intracellular Ca2+ concentration. This is due either to the entry of Ca2+ via P2X purinoceptors or to the activation of metabotropic P2Y purinoceptors followed by signaling via the G protein/phospholipase C/inositol 1,4,5-trisphosphate (IP3) cascade. Eventually, IP3 releases Ca2+ from its intracellular pools. 4. There is no convincing evidence for the presence of P2U purinoceptors sensitive to both ATP and UTP, or pyrimidinoceptors sensitive to UTP only, in the central nervous system (CNS). 5. ATP-sensitive P2X and P2Y purinoceptors show a wide distribution in the CNS and appear to regulate important neuronal functions.

  10. Regulation of extracellular ATP in human erythrocytes infected with Plasmodium falciparum.

    Directory of Open Access Journals (Sweden)

    Cora Lilia Alvarez

    Full Text Available In human erythrocytes (h-RBCs various stimuli induce increases in [cAMP] that trigger ATP release. The resulting pattern of extracellular ATP accumulation (ATPe kinetics depends on both ATP release and ATPe degradation by ectoATPase activity. In this study we evaluated ATPe kinetics from primary cultures of h-RBCs infected with P. falciparum at various stages of infection (ring, trophozoite and schizont stages. A "3V" mixture containing isoproterenol (β-adrenergic agonist, forskolin (adenylate kinase activator and papaverine (phosphodiesterase inhibitor was used to induce cAMP-dependent ATP release. ATPe kinetics of r-RBCs (ring-infected RBCs, t-RBCs (trophozoite-infected RBCs and s-RBCs (schizont-infected RBCs showed [ATPe] to peak acutely to a maximum value followed by a slower time dependent decrease. In all intraerythrocytic stages, values of ΔATP1 (difference between [ATPe] measured 1 min post-stimulus and basal [ATPe] increased nonlinearly with parasitemia (from 2 to 12.5%. Under 3V exposure, t-RBCs at parasitemia 94% (t94-RBCs showed 3.8-fold higher ΔATP1 values than in h-RBCs, indicative of upregulated ATP release. Pre-exposure to either 100 µM carbenoxolone, 100 nM mefloquine or 100 µM NPPB reduced ΔATP1 to 83-87% for h-RBCs and 63-74% for t94-RBCs. EctoATPase activity, assayed at both low nM concentrations (300-900 nM and 500 µM exogenous ATPe concentrations increased approx. 400-fold in t94-RBCs, as compared to h-RBCs, while intracellular ATP concentrations of t94-RBCs were 65% that of h-RBCs. In t94-RBCs, production of nitric oxide (NO was approx. 7-fold higher than in h-RBCs, and was partially inhibited by L-NAME pre-treatment. In media with L-NAME, ΔATP1 values were 2.7-times higher in h-RBCs and 4.2-times higher in t94-RBCs, than without L-NAME. Results suggest that P. falciparum infection of h-RBCs strongly activates ATP release via Pannexin 1 in these cells. Several processes partially counteracted ATPe accumulation: an

  11. ATP storage and uptake by isolated pancreatic zymogen granules

    DEFF Research Database (Denmark)

    Haanes, Kristian Agmund; Novak, Ivana

    2010-01-01

    ATP is released from pancreatic acini in response to cholinergic and hormonal stimulation. The same stimuli cause exocytosis of ZG (zymogen granules) and release of digestive enzymes. The aim of the present study was to determine whether ZG stored ATP and to characterize the uptake mechanism...... for ATP transport into the ZG. ZG were isolated and the ATP content was measured using luciferin/luciferase assays and was related to protein in the sample. The estimate of ATP concentration in freshly isolated granules was 40-120 µM. The ATP uptake had an apparent Km value of 4.9±2.1 mM when granules...... were incubated without Mg2+ and a Km value of 0.47±0.05 mM in the presence of Mg2+, both in pH 6.0 buffers. The uptake of ATP was significantly higher at pH 7.2 compared with pH 6.0 solutions. The anion transport blockers DIDS (4,4'-di-isothiocyanostilbene-2,2'-disulfonate) and Evans Blue inhibited ATP...

  12. K ATP channels in pig and human intracranial arteries

    DEFF Research Database (Denmark)

    Ploug, Kenneth Beri; Sørensen, Mette Aaskov; Strøbech, Lotte Bjørg

    2008-01-01

    Clinical trials suggest that synthetic ATP-sensitive K(+) (K(ATP)) channel openers may cause headache and migraine by dilating cerebral and meningeal arteries. We studied the mRNA expression profile of K(ATP) channel subunits in the pig and human middle meningeal artery (MMA) and in the pig middle...... cerebral artery (MCA). We determined the order of potency of four K(ATP) channel openers when applied to isolated pig MMA and MCA, and we examined the potential inhibitory effects of the Kir6.1 subunit specific K(ATP) channel blocker PNU-37883A on K(ATP) channel opener-induced relaxation of the isolated...... pig MMA and MCA. Using conventional RT-PCR, we detected the mRNA transcripts of the K(ATP) channel subunits Kir6.1 and SUR2B in all the examined pig and human intracranial arteries. Application of K(ATP) channel openers to isolated pig MMA and MCA in myographs caused a concentration...

  13. Highly selective and sensitive electrochemical biosensor for ATP based on the dual strategy integrating the cofactor-dependent enzymatic ligation reaction with self-cleaving DNAzyme-amplified electrochemical detection.

    Science.gov (United States)

    Lu, Lu; Si, Jing Cao; Gao, Zhong Feng; Zhang, Yu; Lei, Jing Lei; Luo, Hong Qun; Li, Nian Bing

    2015-01-15

    A dual strategy that combines the adenosine triphosphate (ATP)-dependent enzymatic ligation reaction with self-cleaving DNAzyme-amplified electrochemical detection is employed to construct the biosensor. In this design, the methylene blue-labeled hairpin-structured DNA was self-assembled onto a gold electrode surface to prepare the modified electrode through the interaction of Au-S bond. In the procedure of ATP-dependent ligation reaction, when the specific cofactor ATP was added, the two split oligonucleotide fragments of 8-17 DNAzyme were linked by T4 DNA ligase and then released to hybridize with the labeled hairpin-structured DNA substrate. The linked 8-17 DNAzyme catalyzes the cleavage of the hairpin-structured substrate by the addition of Zn(2+), causing the methylene blue which contains high electrochemical activity to leave the surface of the gold electrode, therefore generating a dramatic decrease of electrochemical signal. The decrease of peak current was readily measured by square wave voltammetry and a relatively low detection limit (0.05 nM) was obtained with a linear response range from 0.1 to 1000 nM. By taking advantage of the highly specific cofactor dependence of the DNA ligation reaction, the proposed ligation-induced DNAzyme cascades demonstrate ultrahigh selectivity toward the target cofactor ATP. A catalytic and molecular beacons strategy is further adopted to amplify the electrochemical signal detection achieved by cycling and regenerating the 8-17 DNAzyme to realize enzymatic multiple turnover, thus one DNAzyme can catalyze the cleavage of several hairpin-structured substrates, which improves the sensitivity of the newly designed electrochemical sensing system.

  14. Modelling acceptance of sunlight in high and low photovoltaic concentration

    Energy Technology Data Exchange (ETDEWEB)

    Leutz, Ralf, E-mail: ralf.leutz@leopil.com [Leutz Optics and Illumination UG (haftungsbeschränkt), Marburg (Germany)

    2014-09-26

    A simple model incorporating linear radiation characteristics, along with the optical trains and geometrical concentration ratios of solar concentrators is presented with performance examples for optical trains of HCPV, LCPV and benchmark flat-plate PV.

  15. Design philosophy and construction of a high concentration compound parabolic concentrator

    CSIR Research Space (South Africa)

    Roos, TH

    2010-09-01

    Full Text Available A compound parabolic concentrator (CPC) with a concentration ratio of 16:1 is under development at CSIR for volumetric receiver and solar fuels development. The ideal shape has been approximated by 6 and 12 facets in the longitudinal...

  16. ATP promotes cell survival via regulation of cytosolic [Ca2+] and Bcl-2/Bax ratio in lung cancer cells.

    Science.gov (United States)

    Song, Shanshan; Jacobson, Krista N; McDermott, Kimberly M; Reddy, Sekhar P; Cress, Anne E; Tang, Haiyang; Dudek, Steven M; Black, Stephen M; Garcia, Joe G N; Makino, Ayako; Yuan, Jason X-J

    2016-01-15

    Adenosine triphosphate (ATP) is a ubiquitous extracellular messenger elevated in the tumor microenvironment. ATP regulates cell functions by acting on purinergic receptors (P2X and P2Y) and activating a series of intracellular signaling pathways. We examined ATP-induced Ca(2+) signaling and its effects on antiapoptotic (Bcl-2) and proapoptotic (Bax) proteins in normal human airway epithelial cells and lung cancer cells. Lung cancer cells exhibited two phases (transient and plateau phases) of increase in cytosolic [Ca(2+)] ([Ca(2+)]cyt) caused by ATP, while only the transient phase was observed in normal cells. Removal of extracellular Ca(2+) eliminated the plateau phase increase of [Ca(2+)]cyt in lung cancer cells, indicating that the plateau phase of [Ca(2+)]cyt increase is due to Ca(2+) influx. The distribution of P2X (P2X1-7) and P2Y (P2Y1, P2Y2, P2Y4, P2Y6, P2Y11) receptors was different between lung cancer cells and normal cells. Proapoptotic P2X7 was nearly undetectable in lung cancer cells, which may explain why lung cancer cells showed decreased cytotoxicity when treated with high concentration of ATP. The Bcl-2/Bax ratio was increased in lung cancer cells following treatment with ATP; however, the antiapoptotic protein Bcl-2 demonstrated more sensitivity to ATP than proapoptotic protein Bax. Decreasing extracellular Ca(2+) or chelating intracellular Ca(2+) with BAPTA-AM significantly inhibited ATP-induced increase in Bcl-2/Bax ratio, indicating that a rise in [Ca(2+)]cyt through Ca(2+) influx is the critical mediator for ATP-mediated increase in Bcl-2/Bax ratio. Therefore, despite high ATP levels in the tumor microenvironment, which would induce cell apoptosis in normal cells, the decreased P2X7 and elevated Bcl-2/Bax ratio in lung cancer cells may enable tumor cells to survive. Increasing the Bcl-2/Bax ratio by exposure to high extracellular ATP may, therefore, be an important selective pressure promoting transformation and cancer progression. Copyright

  17. High-performance deployable structures for the support of high-concentration ratio solar array modules

    Science.gov (United States)

    Mobrem, M.

    1985-01-01

    A study conducted on high-performance deployable structures for the support of high-concentration ratio solar array modules is discussed. Serious consideration is being given to the use of high-concentration ratio solar array modules or applications such as space stations. These concentrator solar array designs offer the potential of reduced cost, reduced electrical complexity, higher power per unit area, and improved survivability. Arrays of concentrators, such as the miniaturized Cassegrainian concentrator modules, present a serious challenge to the structural design because their mass per unit area (5.7 kg/square meters) is higher than that of flexible solar array blankets, and the requirement for accurate orientation towards the Sun (plus or minus 0.5 degree) requires structures with improved accuracy potentials. In addition, use on a space station requires relatively high structural natural frequencies to avoid deleterious interactions with control systems and other large structural components. The objective here is to identify and evaluate conceptual designs of structures suitable for deploying and accurately supporting high-concentration ratio solar array modules.

  18. Real time imaging of live cell ATP leaking or release events by chemiluminescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yun [Iowa State Univ., Ames, IA (United States)

    2008-12-18

    The purpose of this research was to expand the chemiluminescence microscopy applications in live bacterial/mammalian cell imaging and to improve the detection sensitivity for ATP leaking or release events. We first demonstrated that chemiluminescence (CL) imaging can be used to interrogate single bacterial cells. While using a luminometer allows detecting ATP from cell lysate extracted from at least 10 bacterial cells, all previous cell CL detection never reached this sensitivity of single bacteria level. We approached this goal with a different strategy from before: instead of breaking bacterial cell membrane and trying to capture the transiently diluted ATP with the firefly luciferase CL assay, we introduced the firefly luciferase enzyme into bacteria using the modern genetic techniques and placed the CL reaction substrate D-luciferin outside the cells. By damaging the cell membrane with various antibacterial drugs including antibiotics such as Penicillins and bacteriophages, the D-luciferin molecules diffused inside the cell and initiated the reaction that produces CL light. As firefly luciferases are large protein molecules which are retained within the cells before the total rupture and intracellular ATP concentration is high at the millmolar level, the CL reaction of firefly luciferase, ATP and D-luciferin can be kept for a relatively long time within the cells acting as a reaction container to generate enough photons for detection by the extremely sensitive intensified charge coupled device (ICCD) camera. The result was inspiring as various single bacterium lysis and leakage events were monitored with 10-s temporal resolution movies. We also found a new way of enhancing diffusion D-luciferin into cells by dehydrating the bacteria. Then we started with this novel single bacterial CL imaging technique, and applied it for quantifying gene expression levels from individual bacterial cells. Previous published result in single cell gene expression quantification

  19. Design and development of a high-concentration and high-efficiency photovoltaic concentrator using a curved Fresnel lens

    Energy Technology Data Exchange (ETDEWEB)

    Scharlack, R.S.; Moffat, A.

    1983-08-01

    Thermo Electron has designed a high concentration photovoltaic module that uses a domed, point-focus Fresnel lens. Their design, design optimization process, and results from lens and receiver tests are described in this report. A complete module has not been fabricated and probably will not be fabricated in the future; however, Thermo Electron's optical design, analysis, and testing of both secondary optical units and domed Fresnel lenses have made a significant contribution to our project. Tooling errors prevented the lens from reaching its potential efficiency by the end of the contract, and resolution of these tooling problems is currently being attempted with a follow-on contract, No. 68-9463.

  20. Cloning, characterization and mapping of the human ATP5E gene, identification of pseudogene ATP5EP1, and definition of the ATP5E motif.

    Science.gov (United States)

    Tu, Q; Yu, L; Zhang, P; Zhang, M; Zhang, H; Jiang, J; Chen, C; Zhao, S

    2000-04-01

    A cDNA encoding the epsilon subunit of human ATP synthase, ATP5E, was isolated from heart, skeletal muscle and spleen cDNA libraries respectively. Its genome structure was characterized as comprising three exons and two introns within a stretch of 5 kb, according to the genomic sequence AL109840. The gene was mapped to human chromosome 20q13.3 between marker D20S173 and 20qter using the radiation hybrid GB4 panel. Northern blot analysis showed that the ATP5E gene was expressed as a single 0.6 kb transcript in all 16 human tissues tested, with a high level present in heart and skeletal muscle. A new conserved motif composed of 24 residues, termed the ATP5E motif [W(R/K)X(5)YX(2)(Y/F)X(3)(C/A)X(4)RX(3)K], was defined on the basis of sequences of ATP synthase epsilon subunits from ten different organisms. In addition, a pseudogene ATP5EP1 was also identified on the basis of genomic sequence AC004066, localized on human chromosome 4q25. By analysing these results combined with the Southern blot patterns of human DNA hybridized with bovine ATP5E cDNA reported previously [Vinas, Powell, Runswick, Iacobazzi and Walker (1990) Biochem. J. 265, 321-326], we provide evidence of yet further homologous sequences (either gene or pseudogene) of ATP5E, in addition to ATP5E and ATP5EP1 in the human genome.

  1. Structure of ATP-Bound Human ATP:Cobalamin Adenosyltransferase

    Energy Technology Data Exchange (ETDEWEB)

    Schubert,H.; Hill, C.

    2006-01-01

    Mutations in the gene encoding human ATP:cobalamin adenosyltransferase (hATR) can result in the metabolic disorder known as methylmalonic aciduria (MMA). This enzyme catalyzes the final step in the conversion of cyanocobalamin (vitamin B{sub 12}) to the essential human cofactor adenosylcobalamin. Here we present the 2.5 {angstrom} crystal structure of ATP bound to hATR refined to an R{sub free} value of 25.2%. The enzyme forms a tightly associated trimer, where the monomer comprises a five-helix bundle and the active sites lie on the subunit interfaces. Only two of the three active sites within the trimer contain the bound ATP substrate, thereby providing examples of apo- and substrate-bound-active sites within the same crystal structure. Comparison of the empty and occupied sites indicates that twenty residues at the enzyme's N-terminus become ordered upon binding of ATP to form a novel ATP-binding site and an extended cleft that likely binds cobalamin. The structure explains the role of 20 invariant residues; six are involved in ATP binding, including Arg190, which hydrogen bonds to ATP atoms on both sides of the scissile bond. Ten of the hydrogen bonds are required for structural stability, and four are in positions to interact with cobalamin. The structure also reveals how the point mutations that cause MMA are deficient in these functions.

  2. Light- and metabolism-related regulation of the chloroplast ATP synthase has distinct mechanisms and functions.

    Science.gov (United States)

    Kohzuma, Kaori; Dal Bosco, Cristina; Meurer, Jörg; Kramer, David M

    2013-05-01

    The chloroplast CF0-CF1-ATP synthase (ATP synthase) is activated in the light and inactivated in the dark by thioredoxin-mediated redox modulation of a disulfide bridge on its γ subunit. The activity of the ATP synthase is also fine-tuned during steady-state photosynthesis in response to metabolic changes, e.g. altering CO2 levels to adjust the thylakoid proton gradient and thus the regulation of light harvesting and electron transfer. The mechanism of this fine-tuning is unknown. We test here the possibility that it also involves redox modulation. We found that modifying the Arabidopsis thaliana γ subunit by mutating three highly conserved acidic amino acids, D211V, E212L, and E226L, resulted in a mutant, termed mothra, in which ATP synthase which lacked light-dark regulation had relatively small effects on maximal activity in vivo. In situ equilibrium redox titrations and thiol redox-sensitive labeling studies showed that the γ subunit disulfide/sulfhydryl couple in the modified ATP synthase has a more reducing redox potential and thus remains predominantly oxidized under physiological conditions, implying that the highly conserved acidic residues in the γ subunit influence thiol redox potential. In contrast to its altered light-dark regulation, mothra retained wild-type fine-tuning of ATP synthase activity in response to changes in ambient CO2 concentrations, indicating that the light-dark- and metabolic-related regulation occur through different mechanisms, possibly via small molecule allosteric effectors or covalent modification.

  3. Carbon Nanofibers Modified Graphite Felt for High Performance Anode in High Substrate Concentration Microbial Fuel Cells

    Directory of Open Access Journals (Sweden)

    Youliang Shen

    2014-01-01

    Full Text Available Carbon nanofibers modified graphite fibers (CNFs/GF composite electrode was prepared for anode in high substrate concentration microbial fuel cells. Electrochemical tests showed that the CNFs/GF anode generated a peak current density of 2.42 mA cm−2 at a low acetate concentration of 20 mM, which was 54% higher than that from bare GF. Increase of the acetate concentration to 80 mM, in which the peak current density of the CNFs/GF anode greatly increased and was up to 3.57 mA cm−2, was seven times as that of GF anode. Morphology characterization revealed that the biofilms in the CNFs/GF anode were much denser than those in the bare GF. This result revealed that the nanostructure in the anode not only enhanced current generation but also could tolerate high substrate concentration.

  4. An Automatic High Efficient Method for Dish Concentrator Alignment

    OpenAIRE

    Yong Wang; Song Li; Jinshan Xu; Yijiang Wang; Xu Cheng; Changgui Gu; Shengyong Chen; Bin Wan

    2014-01-01

    Alignment of dish concentrator is a key factor to the performance of solar energy system. We propose a new method for the alignment of faceted solar dish concentrator. The isosceles triangle configuration of facet’s footholds determines a fixed relation between light spot displacements and foothold movements, which allows an automatic determination of the amount of adjustments. Tests on a 25 kW Stirling Energy System dish concentrator verify the feasibility, accuracy, and efficiency of our...

  5. Expression of ATP7B in normal human liver

    Directory of Open Access Journals (Sweden)

    D Fanni

    2009-06-01

    Full Text Available ATP7B is a copper transporting P-type ATPase, also known as Wilson disease protein, which plays a key role in copper distribution inside cells. Recent experimental data in cell culture have shown that ATP7B putatively serves a dual function in hepatocytes: when localized to the Golgi apparatus, it has a biosynthetic role, delivering copper atoms to apoceruloplasmin; when the hepatocytes are under copper stress, ATP7B translocates to the biliary pole to transport excess copper out of the cell and into the bile canaliculus for subsequent excretion from the body via the bile. The above data on ATP7B localization have been mainly obtained in tumor cell systems in vitro. The aim of the present work was to assess the presence and localization of the Wilson disease protein in the human liver. We tested immunoreactivity for ATP7B in 10 human liver biopsies, in which no significant pathological lesion was found using a polyclonal antiserum specific for ATP7B. In the normal liver, immunoreactivity for ATP7B was observed in hepatocytes and in biliary cells. In the hepatocytes, immunoreactivity for ATP7B was observed close to the plasma membrane, both at the sinusoidal and at the biliary pole. In the biliary cells, ATP7B was localized close to the cell membrane, mainly concentrated at the basal pole of the cells. The data suggest that, in human liver, ATP7B is localized to the plasma membrane of both hepatocytes and biliary epithelial cells.

  6. Selection of common bean lines with high grain yield and high grain calcium and iron concentrations

    Directory of Open Access Journals (Sweden)

    Nerinéia Dalfollo Ribeiro

    2014-02-01

    Full Text Available Genetic improvement of common bean nutritional quality has advantages in marketing and can contribute to society as a food source. The objective of this study was to evaluate the genetic variability for grain yield, calcium and iron concentrations in grains of inbred common bean lines obtained by different breeding methods. For this, 136 F7 inbred lines were obtained using the Pedigree method and 136 F7 inbred lines were obtained using the Single-Seed Descent (SSD method. The lines showed genetic variability for grain yield, and concentrations of calcium and iron independently of the method of advancing segregating populations. The Pedigree method allows obtaining a greater number of lines with high grain yield. Selection using the SSD method allows the identification of a larger number of lines with high concentrations of calcium and iron in grains. Weak negative correlations were found between grain yield and calcium concentration (r = -0.0994 and grain yield and iron concentration (r = -0.3926. Several lines show genetic superiority for grain yield and concentrations of calcium and iron in grains and their selection can result in new common bean cultivars with high nutritional quality.

  7. An Automatic High Efficient Method for Dish Concentrator Alignment

    Directory of Open Access Journals (Sweden)

    Yong Wang

    2014-01-01

    for the alignment of faceted solar dish concentrator. The isosceles triangle configuration of facet’s footholds determines a fixed relation between light spot displacements and foothold movements, which allows an automatic determination of the amount of adjustments. Tests on a 25 kW Stirling Energy System dish concentrator verify the feasibility, accuracy, and efficiency of our method.

  8. Fine-tuned ATP signals are acute mediators in osteocyte mechanotransduction

    DEFF Research Database (Denmark)

    Kringelbach, Tina M.; Aslan, Derya; Novak, Ivana

    2015-01-01

    effects on bone remodeling. Therefore, we hypothesized that ATP signaling is also applied by osteocytes in mechanotransduction. We applied a short fluid pulse on MLO-Y4 osteocyte-like cells during real-time detection of ATP and demonstrated that mechanical stimulation activates the acute release of ATP...... and that these acute ATP signals are fine-tuned according to the magnitude of loading. ATP release was then challenged by pharmacological inhibitors, which indicated a vesicular release pathway for acute ATP signals. Finally, we showed that osteocytes express functional P2X2 and P2X7 receptors and respond to even low...... concentrations of nucleotides by increasing intracellular calcium concentration. These results indicate that in osteocytes, vesicular ATP release is an acute mediator of mechanical signals and the magnitude of loading. These and previous results, therefore, implicate purinergic signaling as an early signaling...

  9. Starch source in high concentrate rations does not affect rumen pH, histamine and lipopolysaccharide concentrations in dairy cows

    NARCIS (Netherlands)

    Pilachai, R.; Schonewille, J.T.; Thamrongyoswittayakul, C.; Aiumlamai, S.; Wachirapakom, C.; Everts, H.; Hendriks, W.H.

    2012-01-01

    The replacement of ground corn by cassava meal on rumen pH, lipopolysaccharide (LPS) and histamine concentrations under typical Thai feeding conditions (high concentrate diets and rice straw as the sole source of roughage) was investigated. Four rumen-fistulated crossbred Holstein, non-pregnant, dry

  10. Bandgap Engineering in High-Efficiency Multijunction Concentrator Cells

    Energy Technology Data Exchange (ETDEWEB)

    King, R. R.; Sherif, R. A.; Kinsey, G. S.; Kurtz, S.; Fetzer, C. M.; Edmondson, K. M.; Law, D. C.; Cotal, H. L.; Krut, D. D.; Ermer, J. H.; Karam, N. H.

    2005-08-01

    This paper discusses semiconductor device research paths under investigation with the aim of reaching the milestone efficiency of 40%. A cost analysis shows that achieving very high cell efficiencies is crucial for the realization of cost-effective photovoltaics, because of the strongly leveraging effect of efficiency on module packaging and balance-of systems costs. Lattice-matched (LM) GaInP/ GaInAs/ Ge 3-junction cells have achieved the highest independently confirmed efficiency at 175 suns, 25?C, of 37.3% under the standard AM1.5D, low-AOD terrestrial spectrum. Lattice-mismatched, or metamorphic (MM), materials offer still higher potential efficiencies, if the crystal quality can be maintained. Theoretical efficiencies well over 50% are possible for a MM GaInP/ 1.17-eV GaInAs/ Ge 3-junction cell limited by radiative recombination at 500 suns. The bandgap - open circuit voltage offset, (Eg/q) - Voc, is used as a valuable theoretical and experimental tool to characterize multijunction cells with subcell bandgaps ranging from 0.7 to 2.1 eV. Experimental results are presented for prototype 6-junction cells employing an active {approx}1.1-eV dilute nitride GaInNAs subcell, with active-area efficiency greater than 23% and over 5.3 V open-circuit voltage under the 1-sun AM0 space spectrum. Such cell designs have theoretical efficiencies under the terrestrial spectrum at 500 suns concentration exceeding 55% efficiency, even for lattice-matched designs.

  11. LPWCO method for the treatment of high concentrated organic wastewater

    Institute of Scientific and Technical Information of China (English)

    1998-01-01

    Based on wet air oxidation (WAO) and Fenton reagent, thispaper raises a new low pressure wet catalytic oxidation(LPWCO)which requires low pressure for the treatment of highlyconcentrated and refractory organic wastewater. Compared withgeneral wet air oxidation, the pressure of the treatment(0.1-0.6MPa) is only one of tens to percentage of latter(3.5-10MPa). Inaddition, its temperature is no more than 180℃.Compared withFenton reagent, while H2O2/COD(weight ratio) less than 1.2, theremoval of COD in the treatment is over twenty percents more thanFenton's even the value of COD is more than 14000mg/L. In thispaper, we study the effect factor of COD removal and the mechanismof this treatment. The existence of synergistic effect (catalytic oxidation and carbonization) for COD removal in H2SO4-Fenton reagent system under the condition of applied pressure and heating (0.1-0.6MPa, 104-165℃) was verified. The best condition of this disposal are as follows:H2O2/COD(weight ratio)=0.2-1.0, Fe2+ 0.6×10-3 mol, H2SO4 0.5mol, COD>1×104mg/L, the operating pressure is 0.1-0.6MPa and temperature is 104-165℃. This method suit to dispose the high-concentrated refractory wastewater, especially to the wastewater containing H2SO produced in the manufacture of pesticide, dyestuff and petrochemical works.

  12. High Oxygen Concentrations Adversely Affect the Performance of Pulmonary Surfactant.

    Science.gov (United States)

    Smallwood, Craig D; Boloori-Zadeh, Parnian; Silva, Maricris R; Gouldstone, Andrew

    2017-08-01

    Although effective in the neonatal population, exogenous pulmonary surfactant has not demonstrated a benefit in pediatric and adult subjects with hypoxic lung injury despite a sound physiologic rationale. Importantly, neonatal surfactant replacement therapy is administered in conjunction with low fractional FIO2 while pediatric/adult therapy is administered with high FIO2 . We suspected a connection between FIO2 and surfactant performance. Therefore, we sought to assess a possible mechanism by which the activity of pulmonary surfactant is adversely affected by direct oxygen exposure in in vitro experiments. The mechanical performance of pulmonary surfactant was evaluated using 2 methods. First, Langmuir-Wilhelmy balance was utilized to study the reduction in surface area (δA) of surfactant to achieve a low bound value of surface tension after repeated compression and expansion cycles. Second, dynamic light scattering was utilized to measure the size of pulmonary surfactant particles in aqueous suspension. For both experiments, comparisons were made between surfactant exposed to 21% and 100% oxygen. The δA of surfactant was 21.1 ± 2.0% and 35.8 ± 2.0% during exposure to 21% and 100% oxygen, respectively (P = .02). Furthermore, dynamic light-scattering experiments revealed a micelle diameter of 336.0 ± 12.5 μm and 280.2 ± 11.0 μm in 21% and 100% oxygen, respectively (P < .001), corresponding to a ∼16% decrease in micelle diameter following exposure to 100% oxygen. The characteristics of pulmonary surfactant were adversely affected by short-term exposure to oxygen. Specifically, surface tension studies revealed that short-term exposure of surfactant film to high concentrations of oxygen expedited the frangibility of pulmonary surfactant, as shown with the δA. This suggests that reductions in pulmonary compliance and associated adverse effects could begin to take effect in a very short period of time. If these findings can be demonstrated in vivo, a role for

  13. High-Efficiency Organic Solar Concentrators for Photovoltaics

    National Research Council Canada - National Science Library

    Michael J. Currie; Jonathan K. Mapel; Timothy D. Heidel; Shalom Goffri; Marc A. Baldo

    2008-01-01

    The cost of photovoltaic power can be reduced with organic solar concentrators. These are planar waveguides with a thin-film organic coating on the face and inorganic solar cells attached to the edges...

  14. ATP and potassium ions: a deadly combination for astrocytes

    Science.gov (United States)

    Jackson, David G.; Wang, Junjie; Keane, Robert W.; Scemes, Eliana; Dahl, Gerhard

    2014-04-01

    The ATP release channel Pannexin1 (Panx1) is self-regulated, i.e. the permeant ATP inhibits the channel from the extracellular space. The affinity of the ATP binding site is lower than that of the purinergic P2X7 receptor allowing a transient activation of Panx1 by ATP through P2X7R. Here we show that the inhibition of Panx1 by ATP is abrogated by increased extracellular potassium ion concentration ([K+]o) in a dose-dependent manner. Since increased [K+]o is also a stimulus for Panx1 channels, it can be expected that a combination of ATP and increased [K+]o would be deadly for cells. Indeed, astrocytes did not survive exposure to these combined stimuli. The death mechanism, although involving P2X7R, does not appear to strictly follow a pyroptotic pathway. Instead, caspase-3 was activated, a process inhibited by Panx1 inhibitors. These data suggest that Panx1 plays an early role in the cell death signaling pathway involving ATP and K+ ions. Additionally, Panx1 may play a second role once cells are committed to apoptosis, since Panx1 is also a substrate of caspase-3.

  15. Microglial migration mediated by ATP-induced ATP release from lysosomes

    Institute of Scientific and Technical Information of China (English)

    Ying Dou; Qing-ming Luo; Shumin Duan; Hang-jun Wu; Hui-quan Li; Song Qin; Yin-er Wang; Jing Li; Hui-fang Lou; Zhong Chen; Xiao-ming Li

    2012-01-01

    Microglia are highly motile cells that act as the main form of active immune defense in the central nervous system.Attracted by factors released from damaged cells,microglia are recruited towards the damaged or infected site,where they are involved in degenerative and regenerative responses and phagocytotic clearance of cell debris.ATP release from damaged neural tissues has been suggested to mediate the rapid extension of microglial process towards the site of injury.However,the mechanisms of the long-range migration of microglia remain to be clarified.Here,we found that lysosomes in microglia contain abundant ATP and exhibit Ca2+-dependent exocytosis in response to various stimuli.By establishing an efficient in vitro chemotaxis assay,we demonstrated that endogenously-released ATP from microglia triggered by local microinjection of ATPγS is critical for the long-range chemotaxis of microglia,a response that was significantly inhibited in microglia treated with an agent inducing iysosome osmodialysis or in cells derived from mice deficient in Rab 27a (ashen mice),a small GTPase required for the trafficking and exocytosis of secretory iysosomes.These results suggest that microglia respond to extracellular ATP by releasing ATP themselves through lysosomal exocytosis,thereby providing a positive feedback mechanism to generate a long-range extracellular signal for attracting distant microglia to migrate towards and accumulate at the site of injury.

  16. 14-3-3 protein is a regulator of the mitochondrial and chloroplast ATP synthase

    OpenAIRE

    Bunney, Tom D.; van Walraven, Hendrika S.; de Boer, Albertus H.

    2001-01-01

    Mitochondrial and chloroplast ATP synthases are key enzymes in plant metabolism, providing cells with ATP, the universal energy currency. ATP synthases use a transmembrane electrochemical proton gradient to drive synthesis of ATP. The enzyme complexes function as miniature rotary engines, ensuring energy coupling with very high efficiency. Although our understanding of the structure and functioning of the synthase has made enormous progress in recent years, our und...

  17. How Native and Alien Metal Cations Bind ATP: Implications for Lithium as a Therapeutic Agent

    Science.gov (United States)

    Dudev, Todor; Grauffel, Cédric; Lim, Carmay

    2017-02-01

    Adenosine triphosphate (ATP), the major energy currency of the cell, exists in solution mostly as ATP-Mg. Recent experiments suggest that Mg2+ interacts with the highly charged ATP triphosphate group and Li+ can co-bind with the native Mg2+ to form ATP-Mg-Li and modulate the neuronal purine receptor response. However, it is unclear how the negatively charged ATP triphosphate group binds Mg2+ and Li+ (i.e. which phosphate group(s) bind Mg2+/Li+) and how the ATP solution conformation depends on the type of metal cation and the metal-binding mode. Here, we reveal the preferred ATP-binding mode of Mg2+/Li+ alone and combined: Mg2+ prefers to bind ATP tridentately to each of the three phosphate groups, but Li+ prefers to bind bidentately to the terminal two phosphates. We show that the solution ATP conformation depends on the cation and its binding site/mode, but it does not change significantly when Li+ binds to Mg2+-loaded ATP. Hence, ATP-Mg-Li, like Mg2+-ATP, can fit in the ATP-binding site of the host enzyme/receptor, activating specific signaling pathways.

  18. How Native and Alien Metal Cations Bind ATP: Implications for Lithium as a Therapeutic Agent

    Science.gov (United States)

    Dudev, Todor; Grauffel, Cédric; Lim, Carmay

    2017-01-01

    Adenosine triphosphate (ATP), the major energy currency of the cell, exists in solution mostly as ATP-Mg. Recent experiments suggest that Mg2+ interacts with the highly charged ATP triphosphate group and Li+ can co-bind with the native Mg2+ to form ATP-Mg-Li and modulate the neuronal purine receptor response. However, it is unclear how the negatively charged ATP triphosphate group binds Mg2+ and Li+ (i.e. which phosphate group(s) bind Mg2+/Li+) and how the ATP solution conformation depends on the type of metal cation and the metal-binding mode. Here, we reveal the preferred ATP-binding mode of Mg2+/Li+ alone and combined: Mg2+ prefers to bind ATP tridentately to each of the three phosphate groups, but Li+ prefers to bind bidentately to the terminal two phosphates. We show that the solution ATP conformation depends on the cation and its binding site/mode, but it does not change significantly when Li+ binds to Mg2+-loaded ATP. Hence, ATP-Mg-Li, like Mg2+-ATP, can fit in the ATP-binding site of the host enzyme/receptor, activating specific signaling pathways. PMID:28195155

  19. Development of manufacturing capability for high-concentration, high-efficiency silicon solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Sinton, R.A.; Verlinden, P.J.; Crane, R.A.; Swanson, R.N. [SunPower Corp., Sunnyvale, CA (United States)

    1996-10-01

    This report presents a summary of the major results from a program to develop a manufacturable, high-efficiency silicon concentrator solar cell and a cost-effective manufacturing facility. The program was jointly funded by the Electric Power Research Institute, Sandia National Laboratories through the Concentrator Initiative, and SunPower Corporation. The key achievements of the program include the demonstration of 26%-efficient silicon concentrator solar cells with design-point (20 W/cm{sup 2}) efficiencies over 25%. High-performance front-surface passivations; that were developed to achieve this result were verified to be absolutely stable against degradation by 475 days of field exposure at twice the design concentration. SunPower demonstrated pilot production of more than 1500 of these cells. This cell technology was also applied to pilot production to supply 7000 17.7-cm{sup 2} one-sun cells (3500 yielded wafers) that demonstrated exceptional quality control. The average efficiency of 21.3% for these cells approaches the peak efficiency ever demonstrated for a single small laboratory cell within 2% (absolute). Extensive cost models were developed through this program and calibrated by the pilot-production project. The production levels achieved indicate that SunPower could produce 7-10 MW of concentrator cells per year in the current facility based upon the cell performance demonstrated during the program.

  20. Highly Sensitive Cadmium Concentration Sensor Using Long Period Grating

    Directory of Open Access Journals (Sweden)

    A. S. Lalasangi

    2011-08-01

    Full Text Available In this paper we have proposed a simple and effective Long Period Grating chemical sensor for detecting the traces of Cadmium (Cd++ in drinking water at ppm level. Long Period gratings (LPG were fabricated by point-by-point technique with CO2 laser. We have characterized the LPG concentration sensor sensitivity for different solutions of Cd concentrations varying from 0.01 ppm to 0.04 ppm by injecting white Light source and observed transmitted spectra using Optical Spectrum Analyzer (OSA. Proper reagents have been used in the solutions for detection of the Cd species. The overall shift in wavelength is 10 nm when surrounding medium gradually changed from water to 0.04 ppm of cadmium concentrations. A comparative study has been done using sophisticated spectroscopic atomic absorption spectrometer (AAS and Inductively Coupled Plasma (ICP instruments. The spectral sensitivity enhancement was done by modifying grating surface with gold nanoparticles.

  1. Methods and devices for high-throughput dielectrophoretic concentration

    Energy Technology Data Exchange (ETDEWEB)

    Simmons, Blake A. (San Francisco, CA); Cummings, Eric B. (Livermore, CA); Fiechtner, Gregory J. (Germantown, MD); Fintschenko, Yolanda (Livermore, CA); McGraw, Gregory J. (Ann Arbor, MI); Salmi, Allen (Escalon, CA)

    2010-02-23

    Disclosed herein are methods and devices for assaying and concentrating analytes in a fluid sample using dielectrophoresis. As disclosed, the methods and devices utilize substrates having a plurality of pores through which analytes can be selectively prevented from passing, or inhibited, on application of an appropriate electric field waveform. The pores of the substrate produce nonuniform electric field having local extrema located near the pores. These nonuniform fields drive dielectrophoresis, which produces the inhibition. Arrangements of electrodes and porous substrates support continuous, bulk, multi-dimensional, and staged selective concentration.

  2. Methods and devices for high-throughput dielectrophoretic concentration

    Science.gov (United States)

    Simmons, Blake A.; Cummings, Eric B.; Fiechtner, Gregory J.; Fintschenko, Yolanda; McGraw, Gregory J.; Salmi, Allen

    2010-02-23

    Disclosed herein are methods and devices for assaying and concentrating analytes in a fluid sample using dielectrophoresis. As disclosed, the methods and devices utilize substrates having a plurality of pores through which analytes can be selectively prevented from passing, or inhibited, on application of an appropriate electric field waveform. The pores of the substrate produce nonuniform electric field having local extrema located near the pores. These nonuniform fields drive dielectrophoresis, which produces the inhibition. Arrangements of electrodes and porous substrates support continuous, bulk, multi-dimensional, and staged selective concentration.

  3. ATP-sulfurylase, sulfur-compounds and plant stress tolerance

    Directory of Open Access Journals (Sweden)

    Naser A. Anjum

    2015-04-01

    Full Text Available Sulfur (S stands fourth in the list of major plant nutrients after N, P and K. Sulfate (SO42-, a form of soil-S taken up by plant roots is metabolically inert. As the first committed step of S-assimilation, ATP-sulfurylase (ATP-S catalyzes SO42--activation and yields activated high-energy compound adenosine-5′-phosphosulfate (APS that is reduced to sulfide (S2- and incorporated into cysteine (Cys. In turn, Cys acts as a precursor or donor of reduced S for a range of S-compounds such as methionine (Met, glutathione (GSH, homo-GSH (h-GSH and phytochelatins (PCs. Among S-compounds, GSH, h-GSH and PCs are known for their involvement in plant tolerance to varied abiotic stresses, Cys is a major component of GSH, h-GSH and PCs; whereas, several key stress-metabolites such as ethylene, are controlled by Met through its first metabolite S-adenosylmethionine. With the major aim of briefly highlighting S-compound-mediated role of ATP-S in plant stress tolerance, this paper: (a overviews ATP-S structure/chemistry and occurrence, (b appraises recent literature available on ATP-S roles and regulations, and underlying mechanisms in plant abiotic and biotic stress tolerance, (c summarizes ATP-S-intrinsic regulation by major S-compounds, and (d highlights major open-questions in the present context. Future research in the current direction can be devised based on the outcomes of the discussion.

  4. Gene expression profiling indicates an increased capacity for proline, serine, and ATP synthesis and mitochondrial mass by the liver of steers grazing high vs. low endophyte-infected tall fescue.

    Science.gov (United States)

    Liao, S F; Boling, J A; Matthews, J C

    2015-12-01

    Grazing -infected forages results in a variety of reduced animal performance parameters, collectively known as "fescue toxicosis." The initial, limited evaluations of hepatic mechanisms affected by fescue toxicosis have used transcriptomic expression profiling of experimental phenotypes developed by short-term feeding of concentrated ergot alkaloids or fescue seeds to rodents and steers. To assess the effects of fescue toxicosis in growing cattle using a commercially relevant phenotype, we induced fescue toxicosis in beef steers by summer-long grazing (89 to 105 d) of a single high toxic endophyte-infected tall fescue pasture (HE; 0.746 μg/g ergot alkaloids; 5.7 ha; = 10; BW = 267 ± 14.5 kg) vs. a low toxic endophyte tall fescue-mixed pasture (LE; 0.023 μg/g ergot alkaloids; 5.7 ha; = 9; BW = 266 ± 10.9 kg). High toxic endophyte tall fescue-mixed pasture steers had decreased BW (313 vs. 338 kg) and an increased potential for hepatic gluconeogenesis from AA-derived carbons. To gain a greater perspective into fescue toxicosis-induced hepatic metabolism and identify candidate regulatory mechanisms, the goal of the current research was to examine liver samples for changes in gene (mRNA) expression profiles using a Bovine Affymetrix microarray and selected reverse-transcription PCR and immunoblot analyses. The expression (false discovery rate endophyte tall fescue-mixed pasture steers had increased ( ≤ 0.022) expression of genes critical for increased (1) Pro () and Ser () synthesis, (2) shunting of AA carbons into pyruvate () and ATP synthesis (, , , COX4, , and ), and (3) mitochondrial mass (COX4). Targeted reverse-transcribed PCR or immunoblot assays corroborated ( ≤ 0.035) these latter microarray findings for , , , , and COX4. Moreover, network analysis identified glucocorticoid receptor-mediated signaling as the most probable mechanism to coordinate the above findings. These results greatly extend our knowledge of the consequences of summer-long grazing of

  5. High Quantum Efficiency and High Concentration Erbium-Doped Silica Glasses Fabricated by Sintering Nanoporous Glasses

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A new method was used to prepare erbium-doped high silica (SiO2%>96%) glasses by sintering nanoporous glasses. The concentration of erbium ions in high silica glasses can be considerably more than that in silica glasses prepared by using conventional methods. The fluorescence of 1532 nm has an FWHM (Full Wave at Half Maximum) of 50 nm, wider than 35 nm of EDSFA (erbium-doped silica fiber amplifer), and hence the glass possesses potential application in broadband fiber amplifiers. The Judd-Ofelt theoretical analysis reflects that the quantum efficiency of this erbium-doped glass is about 0.78, although the erbium concentration in this glass (6×103) is about twenty times higher than that in silica glass. These excellent characteristics of Er-doped high silica glass will be conducive to its usage in optical amplifiers and microchip lasers.

  6. Differential expression of ATP7A, ATP7B and CTR1 in adult rat dorsal root ganglion tissue

    Directory of Open Access Journals (Sweden)

    Ip Virginia

    2010-09-01

    Full Text Available Abstract Background ATP7A, ATP7B and CTR1 are metal transporting proteins that control the cellular disposition of copper and platinum drugs, but their expression in dorsal root ganglion (DRG tissue and their role in platinum-induced neurotoxicity are unknown. To investigate the DRG expression of ATP7A, ATP7B and CTR1, lumbar DRG and reference tissues were collected for real time quantitative PCR, RT-PCR, immunohistochemistry and Western blot analysis from healthy control adult rats or from animals treated with intraperitoneal oxaliplatin (1.85 mg/kg or drug vehicle twice weekly for 8 weeks. Results In DRG tissue from healthy control animals, ATP7A mRNA was clearly detectable at levels similar to those found in the brain and spinal cord, and intense ATP7A immunoreactivity was localised to the cytoplasm of cell bodies of smaller DRG neurons without staining of satellite cells, nerve fibres or co-localisation with phosphorylated heavy neurofilament subunit (pNF-H. High levels of CTR1 mRNA were detected in all tissues from healthy control animals, and strong CTR1 immunoreactivity was associated with plasma membranes and vesicular cytoplasmic structures of the cell bodies of larger-sized DRG neurons without co-localization with ATP7A. DRG neurons with strong expression of ATP7A or CTR1 had distinct cell body size profiles with minimal overlap between them. Oxaliplatin treatment did not alter the size profile of strongly ATP7A-immunoreactive neurons but significantly reduced the size profile of strongly CTR1-immunoreactive neurons. ATP7B mRNA was barely detectable, and no specific immunoreactivity for ATP7B was found, in DRG tissue from healthy control animals. Conclusions In conclusion, adult rat DRG tissue exhibits a specific pattern of expression of copper transporters with distinct subsets of peripheral sensory neurons intensely expressing either ATP7A or CTR1, but not both or ATP7B. The neuron subtype-specific and largely non

  7. Responses of Rat P2X2 Receptors to Ultrashort Pulses of ATP Provide Insights into ATP Binding and Channel Gating

    Science.gov (United States)

    Moffatt, Luciano; Hume, Richard I.

    2007-01-01

    To gain insight into the way that P2X2 receptors localized at synapses might function, we explored the properties of outside-out patches containing many of these channels as ATP was very rapidly applied and removed. Using a new method to calibrate the speed of exchange of solution over intact patches, we were able to reliably produce applications of ATP lasting <200 μs. For all concentrations of ATP, there was a delay of at least 80 μs between the time when ATP arrived at the receptor and the first detectable flow of inward current. In response to 200-μs pulses of ATP, the time constant of the rising phase of the current was ∼600 μs. Thus, most channel openings occurred when no free ATP was present. The current deactivated with a time constant of ∼60 ms. The amplitude of the peak response to a brief pulse of a saturating concentration of ATP was ∼70% of that obtained during a long application of the same concentration of ATP. Thus, ATP leaves fully liganded channels without producing an opening at least 30% of the time. Extensive kinetic modeling revealed three different schemes that fit the data well, a sequential model and two allosteric models. To account for the delay in opening at saturating ATP, it was necessary to incorporate an intermediate closed state into all three schemes. These kinetic properties indicate that responses to ATP at synapses that use homomeric P2X2 receptors would be expected to greatly outlast the duration of the synaptic ATP transient produced by a single presynaptic spike. Like NMDA receptors, P2X2 receptors provide the potential for complex patterns of synaptic integration over a time scale of hundreds of milliseconds. PMID:17664346

  8. Concentration measurement of yeast suspensions using high frequency ultrasound backscattering.

    Science.gov (United States)

    Elvira, Luis; Vera, Pedro; Cañadas, Francisco Jesús; Shukla, Shiva Kant; Montero, Francisco

    2016-01-01

    This work proposes the use of an ultrasound based technique to measure the concentration of yeasts in liquid suspension. This measurement was achieved by the detection and quantification of ultrasonic echoes backscattered by the cells. More specifically, the technique was applied to the detection and quantification of Saccharomyces cerevisiae. A theoretical approach was proposed to get the average density and sound speed of the yeasts, which were found to be 1116 kg/m(3) and 1679 m/s, respectively. These parameters were needed to model the waves backscattered by each single cell. A pulse-echo arrangement working around 50 MHz, being able to detect echoes from single yeasts was used to characterize experimentally yeast solutions from 10(2) to 10(7)cells/ml. The Non-negative Matrix Factorization denoising technique was applied for data analysis. This technique required a previous learning of the spectral patterns of the echoes reflected from yeasts in solution and the base noise from the liquid medium. Comparison between pulse correlation (without denoising) and theoretical and experimental pattern learning was made to select the best signal processing. A linear relation between ultrasound output and concentration was obtained with correlation coefficient R(2)=0.996 for the experimental learning. Concentrations from 10(4) to 10(7)cells/ml were detected above the base noise. These results show the viability of using the ultrasound backscattering technique to detect yeasts and measure their concentration in liquid cultures, improving the sensitivity obtained using spectrophotometric methods by one order of magnitude.

  9. Dimerization in Highly Concentrated Solutions of Phosphoimidazolide Activated Monomucleotides

    Science.gov (United States)

    Kanavarioti, Anastassia

    1997-08-01

    Phosphoimidazolide activated ribomononucleotides (*pN) are useful substrates for the non-enzymatic synthesis of polynucleotides. However, dilute neutral aqueous solutions of *pN typically yield small amounts of dimers and traces of polymers; most of *pN hydrolyzes to yield nucleoside 5'-monophosphate. Here we report the self-condensation of nucleoside 5'-phosphate 2-methylimidazolide (2-MeImpN with N = cytidine, uridine or guanosine) in the presence of Mg2+ in concentrated solutions, such as might have been found in an evaporating lagoon on prebiotic Earth. The product distribution indicates that oligomerization is favored at the expense of hydrolysis. At 1.0 M, 2-MeImpU and 2-MeImpC produce about 65% of oligomers including 4% of the 3',5'-linked dimer. Examination of the product distribution of the three isomeric dimers in a self-condensation allows identification of reaction pathways that lead to dimer formation. Condensations in a concentrated mixture of all three nucleotides (U,C,G mixtures) is made possible by the enhanced solubility of 2-MeImpG in such mixtures. Although percent yield of internucleotide linked dimers is enhanced as a function of initial monomer concentration, pyrophosphate dimer yields remain practically unchanged at about 20% for 2-MeImpU, 16% for 2-MeImpC and 25% of the total pyrophosphate in the U,C,G mixtures. The efficiency by which oligomers are produced in these concentrated solutions makes the evaporating lagoon scenario a potentially interesting medium for the prebiotic synthesis of dimers and short RNAs.

  10. Continuous-flow ATP amplification system for increasing the sensitivity of quantitative bioluminescence assay.

    Science.gov (United States)

    Satoh, Tetsuya; Shinoda, Yasuharu; Alexandrov, Maxym; Kuroda, Akio; Murakami, Yuji

    2008-08-01

    We constructed a novel ATP amplification reactor using a continuous-flow system, and this allowed us to increase the sensitivity of a quantitative bioluminescence assay by controlling the number of ATP amplification cycles. We previously developed a bioluminescence assay coupled with ATP amplification using a batch system. However, it was difficult to control the number of amplification cycles. In this study, ATP amplification was performed using a continuous-flow system, and significant linear correlations between amplified luminescence and initial ATP concentration were observed. When performing four cycles of continuous-flow ATP amplification, the gradient of amplification was 1.87(N). Whereas the lower quantifiable level was 500 pM without amplification, values as low as 50 pM ATP could be measured after amplification. The sensitivity thus increased 10-fold, with further improvements expected with additional amplification cycles. The continuous-flow system thus effectively increased the sensitivity of the quantitative bioluminescence assay.

  11. High concentrations of pepsin in bronchoalveolar lavage fluid from children with cystic fibrosis are associated with high interleukin-8 concentrations.

    LENUS (Irish Health Repository)

    McNally, P

    2012-02-01

    BACKGROUND: Gastro-oesophageal reflux is common in children with cystic fibrosis (CF) and is thought to be associated with pulmonary aspiration of gastric contents. The measurement of pepsin in bronchoalveolar lavage (BAL) fluid has recently been suggested to be a reliable indicator of aspiration. The prevalence of pulmonary aspiration in a group of children with CF was assessed and its association with lung inflammation investigated. METHODS: This was a cross-sectional case-control study. BAL fluid was collected from individuals with CF (n=31) and healthy controls (n=7). Interleukin-8 (IL-8), pepsin, neutrophil numbers and neutrophil elastase activity levels were measured in all samples. Clinical, microbiological and lung function data were collected from medical notes. RESULTS: The pepsin concentration in BAL fluid was higher in the CF group than in controls (mean (SD) 24.4 (27.4) ng\\/ml vs 4.3 (4.0) ng\\/ml, p=0.03). Those with CF who had raised pepsin concentrations had higher levels of IL-8 in the BAL fluid than those with a concentration comparable to controls (3.7 (2.7) ng\\/ml vs 1.4 (0.9) ng\\/ml, p=0.004). Within the CF group there was a moderate positive correlation between pepsin concentration and IL-8 in BAL fluid (r=0.48, p=0.04). There was no association between BAL fluid pepsin concentrations and age, sex, body mass index z score, forced expiratory volume in 1 s or Pseudomonas aeruginosa colonisation status. CONCLUSIONS: Many children with CF have increased levels of pepsin in the BAL fluid compared with normal controls. Increased pepsin levels were associated with higher IL-8 concentrations in BAL fluid. These data suggest that aspiration of gastric contents occurs in a subset of patients with CF and is associated with more pronounced lung inflammation.

  12. High pressure inertial focusing for separating and concentrating bacteria at high throughput

    Science.gov (United States)

    Cruz, J.; Hooshmand Zadeh, S.; Graells, T.; Andersson, M.; Malmström, J.; Wu, Z. G.; Hjort, K.

    2017-08-01

    Inertial focusing is a promising microfluidic technology for concentration and separation of particles by size. However, there is a strong correlation of increased pressure with decreased particle size. Theory and experimental results for larger particles were used to scale down the phenomenon and find the conditions that focus 1 µm particles. High pressure experiments in robust glass chips were used to demonstrate the alignment. We show how the technique works for 1 µm spherical polystyrene particles and for Escherichia coli, not being harmful for the bacteria at 50 µl min-1. The potential to focus bacteria, simplicity of use and high throughput make this technology interesting for healthcare applications, where concentration and purification of a sample may be required as an initial step.

  13. Adenosine Triphosphate (ATP Is a Candidate Signaling Molecule in the Mitochondria-to-Nucleus Retrograde Response Pathway

    Directory of Open Access Journals (Sweden)

    Zhengchang Liu

    2013-03-01

    Full Text Available Intracellular communication from the mitochondria to the nucleus is achieved via the retrograde response. In budding yeast, the retrograde response, also known as the RTG pathway, is regulated positively by Rtg1, Rtg2, Rtg3 and Grr1 and negatively by Mks1, Lst8 and two 14-3-3 proteins, Bmh1/2. Activation of retrograde signaling leads to activation of Rtg1/3, two basic helix-loop-helix leucine zipper transcription factors. Rtg1/3 activation requires Rtg2, a cytoplasmic protein with an N-terminal adenosine triphosphate (ATP binding domain belonging to the actin/Hsp70/sugar kinase superfamily. The critical regulatory step of the retrograde response is the interaction between Rtg2 and Mks1. Rtg2 binds to and inactivates Mks1, allowing for activation of Rtg1/3 and the RTG pathway. When the pathway is inactive, Mks1 has dissociated from Rtg2 and bound to Bmh1/2, preventing activation of Rtg1/3. What signals association or disassociation of Mks1 and Rtg2 is unknown. Here, we show that ATP at physiological concentrations dissociates Mks1 from Rtg2 in a highly cooperative fashion. We report that ATP-mediated dissociation of Mks1 from Rtg2 is conserved in two other fungal species, K. lactis and K. waltii. Activation of Rtg1/3 upregulates expression of genes encoding enzymes catalyzing the first three reactions of the Krebs cycle, which is coupled to ATP synthesis through oxidative phosphorylation. Therefore, we propose that the retrograde response is an ATP homeostasis pathway coupling ATP production with ATP-mediated repression of the retrograde response by releasing Mks1 from Rtg2.

  14. New PSA high concentration solar furnace SF40

    Science.gov (United States)

    Rodriguez, Jose; Cañadas, Inmaculada; Zarza, Eduardo

    2016-05-01

    A new solar furnace has been designed and built at Plataforma Solar de Almería. In this work, its main components such as heliostat, concentrator, attenuator and test table, and the method used to align them are described. Other equipment like the auxiliary systems necessary for the solar operation, vacuum chamber and gas system are outlined too. Finally, the thermal characteristics of the focus were measured during a test campaign, where different planes along the optical axis were scanned with a radiometer, and the peak flux was obtained and is presented in the last section of this report.

  15. Evaluation of ATP measurements to detect microbial ingress by wastewater and surface water in drinking water.

    Science.gov (United States)

    Vang, Óluva K; Corfitzen, Charlotte B; Smith, Christian; Albrechtsen, Hans-Jørgen

    2014-11-01

    Fast and reliable methods are required for monitoring of microbial drinking water quality in order to protect public health. Adenosine triphosphate (ATP) was investigated as a potential real-time parameter for detecting microbial ingress in drinking water contaminated with wastewater or surface water. To investigate the ability of the ATP assay in detecting different contamination types, the contaminant was diluted with non-chlorinated drinking water. Wastewater, diluted at 10(4) in drinking water, was detected with the ATP assay, as well as 10(2) to 10(3) times diluted surface water. To improve the performance of the ATP assay in detecting microbial ingress in drinking water, different approaches were investigated, i.e. quantifying microbial ATP or applying reagents of different sensitivities to reduce measurement variations; however, none of these approaches contributed significantly in this respect. Compared to traditional microbiological methods, the ATP assay could detect wastewater and surface water in drinking water to a higher degree than total direct counts (TDCs), while both heterotrophic plate counts (HPC 22 °C and HPC 37 °C) and Colilert-18 (Escherichia coli and coliforms) were more sensitive than the ATP measurements, though with much longer response times. Continuous sampling combined with ATP measurements displays definite monitoring potential for microbial drinking water quality, since microbial ingress in drinking water can be detected in real-time with ATP measurements. The ability of the ATP assay to detect microbial ingress is influenced by both the ATP load from the contaminant itself and the ATP concentration in the specific drinking water. Consequently, a low ATP concentration of the specific drinking water facilitates a better detection of a potential contamination of the water supply with the ATP assay.

  16. High-performance flat-panel solar thermoelectric generators with high thermal concentration.

    Science.gov (United States)

    Kraemer, Daniel; Poudel, Bed; Feng, Hsien-Ping; Caylor, J Christopher; Yu, Bo; Yan, Xiao; Ma, Yi; Wang, Xiaowei; Wang, Dezhi; Muto, Andrew; McEnaney, Kenneth; Chiesa, Matteo; Ren, Zhifeng; Chen, Gang

    2011-05-01

    The conversion of sunlight into electricity has been dominated by photovoltaic and solar thermal power generation. Photovoltaic cells are deployed widely, mostly as flat panels, whereas solar thermal electricity generation relying on optical concentrators and mechanical heat engines is only seen in large-scale power plants. Here we demonstrate a promising flat-panel solar thermal to electric power conversion technology based on the Seebeck effect and high thermal concentration, thus enabling wider applications. The developed solar thermoelectric generators (STEGs) achieved a peak efficiency of 4.6% under AM1.5G (1 kW m(-2)) conditions. The efficiency is 7-8 times higher than the previously reported best value for a flat-panel STEG, and is enabled by the use of high-performance nanostructured thermoelectric materials and spectrally-selective solar absorbers in an innovative design that exploits high thermal concentration in an evacuated environment. Our work opens up a promising new approach which has the potential to achieve cost-effective conversion of solar energy into electricity.

  17. Rebaudioside A directly stimulates insulin secretion from pancreatic beta cells: a glucose-dependent action via inhibition of ATP-sensitive K-channels.

    Science.gov (United States)

    Abudula, R; Matchkov, V V; Jeppesen, P B; Nilsson, H; Aalkjaer, C; Hermansen, K

    2008-11-01

    Recently, we showed that rebaudioside A potently stimulates the insulin secretion from isolated mouse islets in a dose-, glucose- and Ca(2+)-dependent manner. Little is known about the mechanisms underlying the insulinotropic action of rebaudioside A. The aim of this study was to define the signalling system by which, rebaudioside A acts. Isolated mouse islets were used in the cAMP[(125)I] scintillation proximity assay to measure total cAMP level, and in a luminometric method to measure intracellular ATP and ADP concentrations. Conventional and permeabilized whole-cell configuration of the patch-clamp technique was used to verify the effect of rebaudioside A on ATP-sensitive K(+)-channels from dispersed single beta cells from isolated mouse islets. Insulin was measured by radioimmunoassay from insulinoma MIN6 cells. In the presence of 16.7 mM glucose, the addition of the maximally effective concentration of rebaudioside A (10(-9) M) increased the ATP/ADP ratio significantly, while it did not change the intracellular cAMP level. Rebaudioside A (10(-9) M) and stevioside (10(-6) M) reduced the ATP-sensitive potassium channel (K(ATP)) conductance in a glucose-dependent manner. Moreover, rebaudioside A stimulated the insulin secretion from MIN6 cells in a dose- and glucose-dependent manner. In conclusion, the insulinotropic effect of rebaudioside A is mediated via inhibition of ATP-sensitive K(+)-channels and requires the presence of high glucose. The inhibition of ATP-sensitive K(+)-channels is probably induced by changes in the ATP/ADP ratio. The results indicate that rebaudioside A may offer a distinct therapeutic advantage over sulphonylureas because of less risk of causing hypoglycaemia.

  18. Lygus hesperus (Heteroptera: Miridae) tolerates high concentrations of dietary nickel

    Institute of Scientific and Technical Information of China (English)

    ROBERT S. BOYD

    2007-01-01

    Nickel hyperaccumulator plants contain unusually elevated levels of Ni (> 1 000 μg Ni/g). Some insect herbivores, including Lygus hesperus (Western tarnished plant bug), have been observed feeding on the California Ni hyperaccumulator Streptanthus polygaloides. This bug may be able to utilize S. polygaloides as a host either through its feeding behavior or by physiological tolerance of Ni. This experiment determined the Ni tolerance of L. hesperus by offering insects artificial diet amended with 0,0.4, 1, 2, 4.5,10, 20 and 40 mmol Ni/L and recording survival. Survival varied due to Ni concentration, with diets containing 10 mmol Ni/L and greater resulting in significantly lower survival compared to the control (0 mmol Ni/L) treatment. Insects tolerated diet containing as much as 4.5 mmol Ni/L, a relatively elevated Ni concentration. I conclude that L. hesperus can feed on S. polygaloides because it is Ni-tolerant, probably due to physiological mechanisms that provide it with resistance to plant chemical defenses including elemental defenses such as hyperaccumulated Ni.

  19. Adenosine Triphosphate (ATP) Inhibits Voltage-Sensitive Potassium Currents in Isolated Hensen's Cells and Nifedipine Protects Against Noise-Induced Hearing Loss in Guinea Pigs.

    Science.gov (United States)

    Ye, Rui; Liu, Jun; Jia, Zhiying; Wang, Hongyang; Wang, YongAn; Sun, Wei; Wu, Xuan; Zhao, Zhifei; Niu, Baolong; Li, Xingqi; Dai, Guanghai; Li, Jianxiong

    2016-06-13

    BACKGROUND There is increasing evidence that adenosine triphosphate (ATP), a well-known neurotransmitter and neuromodulator in the central nervous system, plays an important role as an extracellular chemical messenger in the cochlea. MATERIAL AND METHODS Using a whole-cell recording technique, we studied the effects of ATP on isolated Hensen's cells, which are supporting cells in the cochlea, to determine if they are involved in the transduction of ions with hair cells. RESULTS ATP (0.1-10 µM) reduced the potassium current (IK+) in the majority of the recorded Hensen's cells (21 out of 25 cells). An inward current was also induced by high concentrations of ATP (100 µM to 10 mM), which was reversibly blocked by 100 µM suramin (a purinergic antagonist) and blocked by nifedipine (an L-type calcium channel blocker). After the cochleas were perfused with artificial perilymph solutions containing nifedipine and exposed to noise, the amplitude increase in the compound action potential (CAP) threshold and the reduction in cochlear microphonics was lower than when they were exposed to noise alone. CONCLUSIONS Our results suggest that ATP can block IK+ channels at a low concentration and induce an inward Ca2+ current at high concentrations, which is reversed by purinergic receptors. Nifedipine may have a partially protective effect on noise-induced hearing loss (NIHL).

  20. Yeast ADP/ATP Carrier Isoform 2

    Science.gov (United States)

    Clémençon, Benjamin; Rey, Martial; Trézéguet, Véronique; Forest, Eric; Pelosi, Ludovic

    2011-01-01

    The mitochondrial ADP/ATP carrier, or Ancp, is a member of the mitochondrial carrier family responsible for exchanging ADP and ATP across the mitochondrial inner membrane. ADP/ATP transport involves Ancp switching between two conformational states. These can be analyzed using specific inhibitors, carboxyatractyloside (CATR) and bongkrekic acid (BA). The high resolution three-dimensional structure of bovine Anc1p (bAnc1p), as a CATR-carrier complex, has been solved. However, because the structure of the BA-carrier complex has not yet been determined, the detailed mechanism of transport remains unknown. Recently, sample processing for hydrogen/deuterium exchange experiments coupled to mass spectrometry was improved, providing novel insights into bAnc1p conformational transitions due to inhibitor binding. In this work we performed both hydrogen/deuterium exchange-mass spectrometry experiments and genetic manipulations. Because these are very difficult to apply with bovine Anc1p, we used Saccharomyces cerevisiae Anc isoform 2 (ScAnc2p). Significant differences in solvent accessibility were observed throughout the amino acid sequence for ScAnc2p complexed to either CATR or BA. Interestingly, in detergent solution, the conformational dynamics of ScAnc2p were dissimilar to those of bAnc1p, in particular for the upper half of the cavity, toward the intermembrane space, and the m2 loop, which is thought to be easily accessible to the solvent from the matrix in bAnc1p. Our study then focused on the methionyl residues of the Ancp signature sequence, RRRMMM. All our results indicate that the methionine cluster is involved in the ADP/ATP transport mechanism and confirm that the Ancp cavity is a highly dynamic structure. PMID:21868387

  1. Influence of the ADP/ATP ratio, 2-oxoglutarate and divalent ions on Azospirillum brasilense PII protein signalling.

    Science.gov (United States)

    Gerhardt, Edileusa C M; Araújo, Luíza M; Ribeiro, Ronny R; Chubatsu, Leda S; Scarduelli, Marcelo; Rodrigues, Thiago E; Monteiro, Rose A; Pedrosa, Fábio O; Souza, Emanuel M; Huergo, Luciano F

    2012-06-01

    Proteins belonging to the P(II) family coordinate cellular nitrogen metabolism by direct interaction with a variety of enzymes, transcriptional regulators and transporters. The sensing function of P(II) relies on its ability to bind the nitrogen/carbon signalling molecule 2-oxoglutarate (2-OG). In Proteobacteria, P(II) is further subject to reversible uridylylation according to the intracellular levels of glutamine, which reflect the cellular nitrogen status. A number of P(II) proteins have been shown to bind ADP and ATP in a competitive manner, suggesting that P(II) might act as an energy sensor. Here, we analyse the influence of the ADP/ATP ratio, 2-OG levels and divalent metal ions on in vitro uridylylation of the Azospirillum brasilense P(II) proteins GlnB and GlnZ, and on interaction with their targets AmtB, DraG and DraT. The results support the notion that the cellular concentration of 2-OG is a key factor governing occupation of the GlnB and GlnZ nucleotide binding sites by ATP or ADP, with high 2-OG levels favouring the occupation of P(II) by ATP. Both P(II) uridylylation and interaction with target proteins responded to the ADP/ATP ratio within the expected physiological range, supporting the concept that P(II) proteins might act as cellular energy sensors.

  2. High Energy Density Lithium Air Batteries for Oxygen Concentrators Project

    Data.gov (United States)

    National Aeronautics and Space Administration — For NASA's Exploration Medical Capabilities mission, extremely high specific energy power sources, with specific energy over 2000 Wh/kg, are urgently sought after....

  3. Changes of IK,ATP current density and allosteric modulation during chronic atrial fibrillation

    Institute of Scientific and Technical Information of China (English)

    WU Gang; HUANG Cong-xin; TANG Yan-hong; JIANG Hong; WAN Jun; CHEN Hui; XIE Qiang; HUANG Zheng-rong

    2005-01-01

    Background Atrial fibrillation (AF) is the most common supraventricular arrhythmia in clinical practice. Chronic atrial fibrillation (CAF) is associated with ionic remodeling. However, little is known about the activity of ATP-sensitive potassium current (IK,ATP) during CAF. So we studied the changes of IK,ATP density and allosteric modulation of ATP-sensitivity by intracellular pH during CAF.Methods Myocardium samples were obtained from the right auricular appendage of patients with rheumatic heart disease complicated with valvular disease in sinus rhythm (SR) or CAF. There were 14 patients in SR group and 9 patients in CAF group. Single atrial cells were isolated using an enzyme dispersion technique. IK,ATP was recorded using the whole-cell and inside-out configuration of voltage-clamp techniques. In whole-cell model, myocytes of SR and CAF groups were perfused with simulated ischemic solution to elicit IK,ATP. In inside-out configuration, the internal patch membranes were exposed to different ATP concentrations in pH 7.4 and 6.8.Results Under simulated ischemia, IK,ATP current density of CAF group was significantly higher than in SR group [(83.5±10.8) vs. (58.7±8.4) pA/pF, P<0.01]. IK,ATP of the two groups showed ATP concentration-dependent inhibition. The ATP concentration for 50% current inhibition (IC50) for the SR group was significantly different in pH 7.4 and pH 6.8 (24 vs. 74 μmol/L, P<0.01). The IC50 did not change significantly in CAF group when the pH decreased from 7.4 to 6.8.Conclusions During CAF, IK,ATP current density was increased and its allosteric modulation of ATP-sensitivity by intracellular pH was diminished.

  4. Celecoxib concentration predicts decrease in prostaglandin E2 concentrations in nipple aspirate fluid from high risk women

    Directory of Open Access Journals (Sweden)

    Flynn John T

    2008-02-01

    Full Text Available Abstract Background Epidemiologic studies suggest that long term low dose celecoxib use significantly lowers breast cancer risk. We previously demonstrated that 400 mg celecoxib taken twice daily for 2 weeks lowered circulating plasma and breast nipple aspirate fluid (NAF prostaglandin (PGE2 concentrations in post- but not premenopausal high risk women. We hypothesized that circulating concentrations of celecoxib influenced PGE2 response, and that plasma levels of the drug are influenced by menopausal status. To address these hypotheses, the aims of the study were to determine: 1 if circulating plasma concentrations of celecoxib correlated with the change in plasma or NAF PGE2 concentrations from baseline to end of treatment, and 2 whether menopausal status influenced circulating levels of celecoxib. Methods Matched NAF and plasma were collected from 46 high risk women who were administered celecoxib twice daily for two weeks, 20 subjects receiving 200 mg and 26 subjects 400 mg of the agent. NAF and plasma samples were collected before and 2 weeks after taking celecoxib. Results In women taking 400 mg bid celecoxib, plasma concentrations of the agent correlated inversely with the change in NAF PGE2 levels from pre- to posttreatment. Nonsignificant trends toward higher celecoxib levels were observed in post- compared to premenopausal women. There was a significant decrease in NAF but not plasma PGE2 concentrations in postmenopausal women who took 400 mg celecoxib (p = 0.03. Conclusion In high risk women taking 400 mg celecoxib twice daily, plasma concentrations of celecoxib correlated with downregulation of PGE2 production by breast tissue. Strategies synergistic with celecoxib to downregulate PGE2 are of interest, in order to minimize the celecoxib dose required to have an effect.

  5. [High efficiency genome walking method for flanking sequences of cotton mitochondrial double-copy atpA gene based on optimized inverse PCR and TAIL-PCR].

    Science.gov (United States)

    Zhang, Xiao; Zhang, Rui; Sun, Guoqing; Shi, Ji; Meng, Zhigang; Zhou, Tao; Hou, Siyu; Liang, Chengzhen; Yu, Yuanhua; Guo, Sandui

    2012-01-01

    Cloning of flanking sequences of double-copy gene is a challenge in molecular biology. We developed a method to solve this problem by combining an optimized inverse PCR (iPCR) with TAIL-PCR. First, Southern blotting analysis was used to determine a proper restriction enzyme that could obtain proper-length restriction fragments that contained the target gene. Then optimized iPCR was performed to amplify the restriction fragments that contained the separated copies of the gene. Based on the obtained sequences, TAIL-PCR was performed to amplify further flanking regions of the gene. With this method, we obtained all of the EcoR I restriction fragments (2.2-5.1 kb) and Hind III restriction fragments (8.5-11.7 kb) of mitochondrial atpA gene in cytoplasmic male sterile (CMS) line and maintainer line of Upland cotton. The results showed that this method was an efficient approach to clone flanking sequences of double-copy gene.

  6. Application of ATP measurements to the microbiological evaluation of a petroleum reservoir

    Energy Technology Data Exchange (ETDEWEB)

    Jones, P.M.

    1981-06-01

    The objective of the work reported in this document was to determine whether the bioluminescent luciferin/luciferase based adenosine triphosphate (ATP) assay could be used as a rapid field tests for determining the presence and numbers of microorganisms in oil field fluids. The ATP-photometric technique employed is based on the ATP-mediated bioluminescent oxidation of firefly luciferin. Light production is stoichiometrically related to ATP concentration; ATP concentration is related to numbers of living organisms present in a sample. Samples used in this study comprised reservoir fluids collected from several Southern California oilfields. Based on experimental evidence, it was concluded that the ATP assay could be profitably applied to Microbially Enhanced Oil Recovery (MEOR) process monitoring and control. The theoretical basis for the assay, field-usage methodologies, and fundamentals of data interpretation are presented to make the document usable as a field manual.

  7. Determination of water quality variables, endotoxin concentration, and enterobacteriaceae concentration and identification in Southern High Plains dairy lagoons

    Science.gov (United States)

    The objectives of this study were to determine the concentration of endotoxin, determine 20 water quality variables, and identify and enumerate fungal and bacterial pathogens from United States southern High Plains dairy lagoons and control lakes during summer and winter. Water samples were collecte...

  8. Multiple di-leucines in the ATP7A copper transporter are required for retrograde trafficking to the trans-Golgi network.

    Science.gov (United States)

    Zhu, Sha; Shanbhag, Vinit; Hodgkinson, Victoria L; Petris, Michael J

    2016-09-01

    The ATP7A protein is a ubiquitous copper-transporting P-type ATPase that is mutated in the lethal pediatric disorder of copper metabolism, Menkes disease. The steady-state location of ATP7A is within the trans-Golgi network (TGN), where it delivers copper to copper-dependent enzymes within the secretory pathway. However, ATP7A constantly cycles between the TGN and the plasma membrane, and in the presence of high copper concentrations, the exocytic arm of this cycling pathway is enhanced to promote a steady-state distribution of ATP7A to post-Golgi vesicles and the plasma membrane. A single di-leucine endocytic motif within the cytosolic carboxy tail of ATP7A (1487LL) was previously shown to be essential for TGN localization by functioning in retrieval from the plasma membrane, however, the requirement of other di-leucine signals in this region has not been fully investigated. While there has been some success in identifying sequence elements within ATP7A required for trafficking and catalysis, progress has been hampered by the instability of the ATP7A cDNA in high-copy plasmids during replication in Escherichia coli. In this study, we find that the use of DNA synthesis to generate silent mutations across the majority of both mouse and human ATP7A open reading frames was sufficient to stabilize these genes in high-copy plasmids, thus permitting the generation of full-length expression constructs. Using the stabilized mouse Atp7a construct, we identify a second di-leucine motif in the carboxy tail of ATP7A (1459LL) as essential for steady-state localization in the TGN by functioning in endosome-to-TGN trafficking. Taken together, these findings demonstrate that multiple di-leucine signals are required for recycling ATP7A from the plasma membrane to the TGN and illustrate the utility of large-scale codon reassignment as a simple and effective approach to circumvent cDNA instability in high-copy plasmids.

  9. The protective effect of overexpressed ATP7B in bone marrow-derived mesenchymal stem cells against copper toxicity%慢病毒介导ATP7B基因转染对骨髓间充质干细胞在高铜环境中生存能力的影响

    Institute of Scientific and Technical Information of China (English)

    邵存华; 陈圣林; 董天赋; 成峰

    2013-01-01

    Objective:To investigate the role of ATP7B overexpression for the survival of bone marrow-derived mesenchymal stem cells (MSCs) of LEC rat in presence of various copper concentrations in vitro.Methods:Bone marrow cells were isolated from the femurs of LEC rat by flushing with rat MSCs growth medium;the lentivirus package system carrying ATP7B/green fluorescent protein (GFP) gene was constructed,and MSCs were infected by the virus.The expression of ATP7B was measured by immunofluorescence,Western blot and Real-time PCR.The SRB assay was used to analyze the viability of MSCs and HepG2 in the presence of various copper concentrations.Results:We successfully built up LEC rat MSCsATP7B which can stably express high level of ATP7B in vitro.In the SRB assay,the viability of MSCsATP7B was significantly higher than that of either MSCsGFP or HepG2 in the presence of high copper concentrations.Conclusion:MSCs of LEC rat rescued by ATP7B can effectively antagonize copper toxicity in vitro,and may represent a novel strategy for therapy of Wilson disease.%目的:探讨慢病毒转染ATP7B基因能否提高骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,MSCs)在高铜环境中的生存能力.方法:全骨髓贴壁法培养Wilson病模型LEC大鼠的MSCs;构建含有ATP7B基因的慢病毒载体pWPT-ATP7B及对照慢病毒载体pWPT-GFP并转染MSCs,获得表达ATP7B基因的MSCsATP7B细胞及对照组MSCsGFP细胞.利用细胞免疫荧光、Real-timePCR、Western blot长期监测ATP7B基因表达;并以HepG2细胞系作为阳性对照,利用SRB法监测给予不同浓度铜离子处理的干细胞增殖情况.结果:MSCs CD90、CD29表达阳性,CD45、CD1 1b表达阴性;细胞免疫荧光、Western blot及RT-PCR显示MSCsATP7B细胞的ATP7B基因能够长期稳定表达;SRB结果显示,高铜环境中MSCsATP7B细胞的生存能力显著高于MSCsGFP细胞及HepG2(P< 0.05).结论:ATP7B基因修正的LEC大鼠MSCs可有效对抗铜蓄积损害,为Wilson

  10. High-Performance Near-Infrared Luminescent Solar Concentrators.

    Science.gov (United States)

    Rondão, Raquel; Frias, Ana R; Correia, Sandra F H; Fu, Lianshe; de Zea Bermudez, Verónica; André, Paulo S; Ferreira, Rute A S; Carlos, Luís D

    2017-04-12

    Luminescent solar concentrators (LSCs) appear as candidates to enhance the performance of photovoltaic (PV) cells and contribute to reduce the size of PV systems, decreasing, therefore, the amount of material needed and thus the cost associated with energy conversion. One way to maximize the device performance is to explore near-infrared (NIR)-emitting centers, resonant with the maximum optical response of the most common Si-based PV cells. Nevertheless, very few examples in the literature demonstrate the feasibility of fabricating LSCs emitting in the NIR region. In this work, NIR-emitting LSCs are reported using silicon 2,3-naphthalocyanine bis(trihexylsilyloxide) (SiNc or NIR775) immobilized in an organic-inorganic tri-ureasil matrix (t-U(5000)). The photophysical properties of the SiNc dye incorporated into the tri-ureasil host closely resembled those of SiNc in tetrahydrofuran solution (an absolute emission quantum yield of ∼0.17 and a fluorescence lifetime of ∼3.6 ns). The LSC coupled to a Si-based PV device revealed an optical conversion efficiency of ∼1.5%, which is among the largest values known in the literature for NIR-emitting LSCs. The LSCs were posteriorly coupled to a Si-based commercial PV cell, and the synergy between the t-U(5000) and SiNc molecules enabled an effective increase in the external quantum efficiency of PV cells, exceeding 20% in the SiNc absorption region.

  11. An astigmatic corrected target-aligned heliostat for high concentration

    Energy Technology Data Exchange (ETDEWEB)

    Zaibel, R.; Dagan, E.; Karni, J. [Solar Research Facility, The Weizmann Institute of Science, Rehovot (Israel); Ries, Harald [Ludwig-Maximilians-Universitaet, Sektion Physik, Munich (Germany)

    1995-05-28

    Conventional heliostats suffer from astigmatism for non-normal incidence. For tangential rays the focal length is shortened while for sagittal rays it is longer than the nominal focal length. Due to this astigmatism it is impossible to produce a sharp image of the sun, and the rays will be spread over a larger area. In order to correct this the heliostat should have different curvature radii along the sagittal and tangential direction in the heliostat plane just like a non axial part of a paraboloid. In conventional heliostats, where the first axis, fixed with respect to the ground, is vertical while the second, fixed with respect to the reflector surface, is horizontal, such an astigmatism correction is not practical because the sagittal and tangential directions rotate with respect to the reflector. We suggest an alternative mount where the first axis is oriented towards the target. The second axis, perpendicular to the first and tangent to the reflector, coincides with the tangential direction. With this mounting sagittal and tangential direction are fixed with respect to the reflector during operation. Therefore a partial astigmatism compensation is possible. We calculate the optimum correction and show the performance of the heliostat. We also show predicted yearly average concentrations

  12. Emulsifier development for high-concentrated reverse emulsions

    OpenAIRE

    Kovalenko, I. L.; V.P. Kuprin

    2016-01-01

    The reverse emulsions have found broad application in ore mining industry as matrixes of emulsion explosive substances and boring washing waters. The defining characteristic of reverse emulsions of industrial explosive substances is the high stability and immunity to crystallization. Aim: The aim of this work is to assess the mechanism of emulsifiers effect like SMO and some PIBSA-derivatives, that are most abundantly used in world practice, and also to develop an effective domestic emulsifie...

  13. High urinary phthalate concentration associated with delayed pubarche in girls

    DEFF Research Database (Denmark)

    Frederiksen, H; Sørensen, K; Mouritsen, A;

    2012-01-01

    Phthalates are a group of chemicals present in numerous consumer products. They have anti-androgenic properties in experimental studies and are suspected to be involved in human male reproductive health problems. A few studies have shown associations between phthalate exposure and changes...... and controls. We demonstrated that delayed pubarche, but not thelarche, was associated with high phthalate excretion in urine samples from 725 healthy school girls, which may suggest anti-androgenic actions of phthalates in our study group of girls....

  14. Deep Sea Memory of High Atmospheric CO2 Concentration

    Science.gov (United States)

    Mathesius, Sabine; Hofmann, Matthias; Caldeira, Ken; Schellnhuber, Hans Joachim

    2015-04-01

    Carbon dioxide removal (CDR) from the atmosphere has been proposed as a powerful measure to mitigate global warming and ocean acidification. Planetary-scale interventions of that kind are often portrayed as "last-resort strategies", which need to weigh in if humankind keeps on enhancing the climate-system stock of CO2. Yet even if CDR could restore atmospheric CO2 to substantially lower concentrations, would it really qualify to undo the critical impacts of past emissions? In the study presented here, we employed an Earth System Model of Intermediate Complexity (EMIC) to investigate how CDR might erase the emissions legacy in the marine environment, focusing on pH, temperature and dissolved oxygen. Against a background of a world following the RCP8.5 emissions path ("business-as-usual") for centuries, we simulated the effects of two massive CDR interventions with CO2 extraction rates of 5 GtC yr-1 and 25 GtC yr-1, respectively, starting in 2250. We found that the 5 GtC yr-1 scheme would have only minor ameliorative influence on the oceans, even after several centuries of application. By way of contrast, the extreme 25 GtC yr-1 scheme eventually leads to tangible improvements. However, even with such an aggressive measure, past CO2 emissions leave a substantial legacy in the marine environment within the simulated period (i.e., until 2700). In summary, our study demonstrates that anthropogenic alterations of the oceans, caused by continued business-as-usual emissions, may not be reversed on a multi-centennial time scale by the most aspirational geoengineering measures. We also found that a transition from the RCP8.5 state to the state of a strong mitigation scenario (RCP2.6) is not possible, even under the assumption of extreme extraction rates (25 GtC yr-1). This is explicitly demonstrated by simulating additional scenarios, starting CDR already in 2150 and operating until the atmospheric CO2 concentration reaches 280 ppm and 180 ppm, respectively. The simulated

  15. High plasma uric acid concentration: causes and consequences

    Directory of Open Access Journals (Sweden)

    de Oliveira Erick

    2012-04-01

    Full Text Available Abstract High plasma uric acid (UA is a precipitating factor for gout and renal calculi as well as a strong risk factor for Metabolic Syndrome and cardiovascular disease. The main causes for higher plasma UA are either lower excretion, higher synthesis or both. Higher waist circumference and the BMI are associated with higher insulin resistance and leptin production, and both reduce uric acid excretion. The synthesis of fatty acids (tryglicerides in the liver is associated with the de novo synthesis of purine, accelerating UA production. The role played by diet on hyperuricemia has not yet been fully clarified, but high intake of fructose-rich industrialized food and high alcohol intake (particularly beer seem to influence uricemia. It is not known whether UA would be a causal factor or an antioxidant protective response. Most authors do not consider the UA as a risk factor, but presenting antioxidant function. UA contributes to > 50% of the antioxidant capacity of the blood. There is still no consensus if UA is a protective or a risk factor, however, it seems that acute elevation is a protective factor, whereas chronic elevation a risk for disease.

  16. ATP/GTP hydrolysis is required for oxazole and thiazole biosynthesis in the peptide antibiotic microcin B17.

    Science.gov (United States)

    Milne, J C; Eliot, A C; Kelleher, N L; Walsh, C T

    1998-09-22

    In the maturation of the Escherichia coli antibiotic Microcin B17, the product of the mcbA gene is modified posttranslationally by the multimeric Microcin synthetase complex (composed of McbB, C, and D) to cyclize four Cys and four Ser residues to four thiazoles and four oxazoles, respectively. The purified synthetase shows an absolute requirement for ATP or GTP in peptide substrate heterocyclization, with GTP one-third as effective as ATP in initial rate studies. The ATPase/GTPase activity of the synthetase complex is conditional in that ADP or GDP formation requires the presence of substrate; noncyclizable versions of McbA bind to synthetase, but do not induce the NTPase activity. The stoichiometry of ATP hydrolysis and heterocycle formation is 5:1 for a substrate that contains two potential sites of modification. However, at high substrate concentrations (>50Km) heterocycle formation is inhibited, while ATPase activity occurs undiminished, consistent with uncoupling of NTP hydrolysis and heterocycle formation at high substrate concentrations. Sequence homology reveals that the McbD subunit has motifs reminiscent of the Walker B box in ATP utilizing enzymes and of motifs found in small G protein GTPases. Mutagenesis of three aspartates to alanine in these motifs (D132, D147, and D199) reduced Microcin B17 production in vivo and heterocycle formation in vitro, suggesting that the 45 kDa McbD has a regulated ATPase/GTPase domain in its N-terminal region necessary for peptide heterocyclization.

  17. [Determination of the antioxidant properties of activators of mitochondrial ATP-dependent potassium channels with the Amplex Red fluorescent indicator].

    Science.gov (United States)

    Murzaeva, S V; Belova, S P; Mironova, G D

    2013-01-01

    The effect of adaptogens-antihypoxants that participate in the activation of mitochondrial ATP-dependent potassium channels (mitoK(ATP)) at the oxidation of the Amplex Red (AR) fluorescent indicator in a peroxidase system was tested. It was shown that Extralife, Hypoxen, taurine, and synthetic antioxidant ionol can be arranged in the following row, according to the fluorescence inhibition activity: Extralife > Hypoxen > > ionol > taurine; their effect was shown to be concentration-dependent. The calculated K(i) value of fluorescence indicators demonstrate fast and slow phases of inhibition of the AR oxidation by Extralife and Hypoxen. The fast phase occurs in the presence of microdoses (0.05-3 microg/mL) of adaptogens and is related to the competition for H2O2, which is in agreement with our previous data on the mitoK(ATP) activation by doses of adaptogens related to the H2O2 consumption. The slow phase is characteristic of high adaptogen and ionol concentrations and is related to the competition for phenoxyl radicals of resorufin formed during AR oxidation. The obtained results allow one to suggest the application of a highly sensitive model peroxidase system with AR for the preliminary testing of compounds activating mitoK(ATP) channels.

  18. The downstream atpE cistron is efficiently translated via its own cis-element in partially overlapping atpB–atpE dicistronic mRNAs in chloroplasts

    OpenAIRE

    Suzuki, Haruka; Kuroda, Hiroshi; Yukawa, Yasushi; Sugiura, Masahiro

    2011-01-01

    The chloroplast atpB and atpE genes encode subunits β and ε of the ATP synthase, respectively. They are co-transcribed as dicistronic mRNAs in flowering plants. An unusual feature is an overlap (AUGA) of the atpB stop codon (UGA) with the atpE start codon (AUG). Hence, atpE translation has been believed to depend on atpB translation (i.e. translational coupling). Using an in vitro translation system from tobacco chloroplasts, we showed that both atpB and atpE cistrons are translated from the ...

  19. Design requirements for high-efficiency high concentration ratio space solar cells

    Science.gov (United States)

    Rauschenbach, H.; Patterson, R.

    1980-01-01

    A miniaturized Cassegrainian concentrator system concept was developed for low cost, multikilowatt space solar arrays. The system imposes some requirements on solar cells which are new and different from those imposed for conventional applications. The solar cells require a circular active area of approximately 4 mm in diameter. High reliability contacts are required on both front and back surfaces. The back area must be metallurgically bonded to a heat sink. The cell should be designed to achieve the highest practical efficiency at 100 AMO suns and at 80 C. The cell design must minimize losses due to nonuniform illumination intensity and nonnormal light incidence. The primary radiation concern is the omnidirectional proton environment.

  20. P2X receptor-mediated ATP purinergic signaling in health and disease

    Directory of Open Access Journals (Sweden)

    Jiang LH

    2012-09-01

    Full Text Available Lin-Hua JiangSchool of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds, United KingdomAbstract: Purinergic P2X receptors are plasma membrane proteins present in a wide range of mammalian cells where they act as a cellular sensor, enabling cells to detect and respond to extracellular adenosine triphosphate (ATP, an important signaling molecule. P2X receptors function as ligand-gated Ca2+-permeable cationic channels that open upon ATP binding to elevate intracellular Ca2+ concentrations and cause membrane depolarization. In response to sustained activation, P2X receptors induce formation of a pore permeable to large molecules. P2X receptors also interact with distinct functional proteins and membrane lipids to form specialized signaling complexes. Studies have provided compelling evidence to show that such P2X receptor-mediated ATP-signaling mechanisms determine and regulate a growing number and diversity of important physiological processes, including neurotransmission, muscle contraction, and cytokine release. There is accumulating evidence to support strong causative relationships of altered receptor expression and function with chronic pain, inflammatory diseases, cancers, and other pathologies or diseases. Numerous high throughput screening drug discovery programs and preclinical studies have thus far demonstrated the proof of concepts that the P2X receptors are druggable targets and selective receptor antagonism is a promising therapeutics approach. This review will discuss the recent progress in understanding the mammalian P2X receptors with respect to the ATP-signaling mechanisms, physiological and pathophysiological roles, and development and preclinical studies of receptor antagonists.Keywords: extracellular ATP, ion channel, large pore, signaling complex, chronic pain, inflammatory diseases

  1. Characterization of blood donors with high haemoglobin concentration

    DEFF Research Database (Denmark)

    Magnussen, K; Hasselbalch, H C; Ullum, H;

    2013-01-01

    Background and Objectives  The literature contains little on the prevalence and causes of high predonation haemoglobin levels among blood donors. This study aimed to characterize and develop an algorithm to manage would-be donors with polycythaemia. Materials and Methods  Between November 2009......, erythropoietin, ferritin, platelet count and leucocyte count, JAK2 V617 and JAK2 exon12 analysis, as well as other routine measurements. Results  Among 46 such donors, 39 had a history of smoking, which contributes to erythrocytosis. Two had PV, five had severe hypertension, one of them because of renal artery...

  2. Waste to Energy Potential - A High Concentration Anaerobic Bioreactor

    Science.gov (United States)

    2012-05-23

    REPORT DATE 23 MAY 2012 2. REPORT TYPE 3. DATES COVERED 00-00-2012 to 00-00-2012 4. TITLE AND SUBTITLE Waste to Energy Potential - A High...and fermentative bacteria break down organic carbon to VFAs Acetogens break down VFAs to CH3CO2 − and H2 + Acetoclastic methanogens break...s -999999 999999 7 481 su -999999 999999 0 .. -999999 999999 HCA8 pti Flo$&-tdgc Tan.\\ feed su -999999 999999 0 .. -999999 999999 A-l>o -999999

  3. Recent testing of secondary concentrators at NREL`s high-flux solar furnace

    Energy Technology Data Exchange (ETDEWEB)

    Jenkins, D.; Winston, R.; O`Gallagher, J. [Univ. of Chicago, IL (United States); Bingham, C.; Lewandowski, A.; Pitts, R.; Scholl, K. [National Renewable Energy Lab., Golden, CO (United States)

    1996-12-31

    Several tests have been completed on new secondary concentrators at the High-Flux Solar Furnace (HFSF) at the National Renewable Energy Laboratory (NREL). The first test measured the power from the exit of a High-Index secondary that achieves an average concentration of 50,000 suns. The second concentrator tested pumped an Nd:YAG laser crystal. The concentrator designs are presented, along with data from on-sun testing at the HFSF.

  4. Follow the ATP: tumor energy production: a perspective.

    Science.gov (United States)

    Oronsky, Bryan T; Oronsky, Neil; Fanger, Gary R; Parker, Christopher W; Caroen, Scott Z; Lybeck, Michelle; Scicinski, Jan J

    2014-01-01

    As early as the 1920s, the eminent physician and chemist, Otto Warburg, nominated for a second Nobel Prize for his work on fermentation, observed that the core metabolic signature of cancer cells is a high glycolytic flux. Warburg averred that the prime mover of cancer is defective mitochondrial respiration, which drives a switch to an alternative energy source, aerobic glycolysis in lieu of Oxidative Phosphorylation (OXPHOS), in an attempt to maintain cellular viability and support critical macromolecular needs. The cell, deprived of mitochondrial ATP production, must reprogram its metabolism as a secondary survival mechanism to maintain sufficient ATP and NADH levels for macromolecule production, membrane integrity and DNA synthesis as well as maintenance of membrane ionic gradients. A time-tested method to identify and disrupt criminal activity is to "follow the money" since the illicit proceeds from crime are required to underwrite it. By analogy, strategies to target cancer involve following and disrupting the flow of ATP and NADH, the energetic and redox "currencies" of the cell, respectively, since the tumor requires high levels of ATP and NADH, not only for metastasis and proliferation, but also, on a more basic level, for survival. Accordingly, four broad ATP reduction strategies to impact and potentially derail cancer energy production are highlighted herein: 1) small molecule energy-restriction mimetic agents (ERMAs) that target various aspects of energy metabolism, 2) reduction of energy 'subsidization' with autophagy inhibitors, 3) acceleration of ATP turnover to increase energy inefficiency, and 4) dietary energy restriction to limit the energy supply.

  5. High-order optical nonlinearities in nanocomposite films dispersed with semiconductor quantum dots at high concentrations

    Science.gov (United States)

    Tomita, Yasuo; Matsushima, Shun-suke; Yamagami, Ryu-ichi; Jinzenji, Taka-aki; Sakuma, Shohei; Liu, Xiangming; Izuishi, Takuya; Shen, Qing

    2017-06-01

    We describe the nonlinear optical properties of inorganic-organic nanocomposite films in which semiconductor CdSe quantum dots as high as 6.8 vol.% are dispersed. Open/closed Z-scan measurements, degenerate multi-wave mixing and femtosecond pump-probe/transient grating measurements are conducted. It is shown that the observed fifth-order optical nonlinearity has the cascaded third-order contribution that becomes prominent at high concentrations of CdSe QDs. It is also shown that there are picosecond-scale intensity-dependent and nanosecond-scale intensity-independent decay components in absorptive and refractive nonlinearities. The former is caused by the Auger process, while the latter comes from the electron-hole recombination process.

  6. EXPERIMENTAL INVESTIGATIONS ON DIFFUSION CHARACTERISTICS OF HIGH CONCENTRATION JET FLOW IN NEAR REGION

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The diffusion characteristics of the high concentration jet flowissued from a round nozzle above the free surface into moving waterbody were experimentally investigated. By means of flow visualization technique, the diffusion behavior of the high concentration jet was observed and analysed, Concentration distribution in the near region was obtained by sampling and quantitative measurements. Experimental results indicate that the diffusion mechanics of the high concentration jet flow are greatly influenced by initial momentum, buoyancy, turbulent structure, ambient current and boundary conditions, showing complicated flow patterns and concentration distribution characteristics that are different from previous results for submerged jets.

  7. Emulsifier development for high-concentrated reverse emulsions

    Directory of Open Access Journals (Sweden)

    I.L. Kovalenko

    2016-05-01

    Full Text Available The reverse emulsions have found broad application in ore mining industry as matrixes of emulsion explosive substances and boring washing waters. The defining characteristic of reverse emulsions of industrial explosive substances is the high stability and immunity to crystallization. Aim: The aim of this work is to assess the mechanism of emulsifiers effect like SMO and some PIBSA-derivatives, that are most abundantly used in world practice, and also to develop an effective domestic emulsifier of reverse emulsions. Materials and methods: Using the semi-dynamic method with use of the reverse stalagmometer it was determined the decreasing in interfacial tension on “water / diesel fuel” border in the presence of 0.5 wt % sorbitan monooleate of various producers. Emulsions with use of the chosen emulsifiers using the dynamic mixer on the basis of monosolution of ammonium nitrate and diesel fuel have been produced. The emulsions have the following composition, wt %: ammonium nitrate – 76.8; water – 15.6; diesel fuel – 6.0; emulsifier – 1.6. Results: By the researches results of the interfacial tension “surfactant water / solution in diesel fuel”, the stability of emulsions using monosolution of ammonium nitrate and the IR spectrums of SMO of various producers it is established that presence in product of impurity of oleic acid, di- and trioleates leads to decreasing in interphase activity, increasing of emulsifier oil solubility and decreasing the resistance of emulsions to crystallization. On the basis of the spectral data analysis it is suggested about possibility of specific interaction on the mechanism of “spectral resonance” between emulsifiers of the PIBSA-MEA, LZX type and crystals nucleus of NH4NO3 ammonium nitrate in dispersed phase of emulsion. Amidation of vegetable oils by monoethanol amine is implemented at the reduced temperatures (90…100 °C. It was proved the availability mainly of fatty acids amides in product

  8. Ecto-nucleoside triphosphate diphosphohydrolase 2 modulates local ATP-induced calcium signaling in human HaCaT keratinocytes.

    Directory of Open Access Journals (Sweden)

    Chia-Lin Ho

    Full Text Available Keratinocytes are the major building blocks of the human epidermis. In many physiological and pathophysiological conditions, keratinocytes release adenosine triphosphate (ATP as an autocrine/paracrine mediator that regulates cell proliferation, differentiation, and migration. ATP receptors have been identified in various epidermal cell types; therefore, extracellular ATP homeostasis likely determines its long-term, trophic effects on skin health. We investigated the possibility that human keratinocytes express surface-located enzymes that modulate ATP concentration, as well as the corresponding receptor activation, in the pericellular microenvironment. We observed that the human keratinocyte cell line HaCaT released ATP and hydrolyzed extracellular ATP. Interestingly, ATP hydrolysis resulted in adenosine diphosphate (ADP accumulation in the extracellular space. Pharmacological inhibition by ARL 67156 or gene silencing of the endogenous ecto-nucleoside triphosphate diphosphohydrolase (NTPDase isoform 2 resulted in a 25% reduction in both ATP hydrolysis and ADP formation. Using intracellular calcium as a reporter, we found that although NTPDase2 hydrolyzed ATP and generated sustainable ADP levels, only ATP contributed to increased intracellular calcium via P2Y2 receptor activation. Furthermore, knocking down NTPDase2 potentiated the nanomolar ATP-induced intracellular calcium increase, suggesting that NTPDase2 globally attenuates nucleotide concentration in the pericellular microenvironment as well as locally shields receptors in the vicinity from being activated by extracellular ATP. Our findings reveal an important role of human keratinocyte NTPDase2 in modulating nucleotide signaling in the extracellular milieu of human epidermis.

  9. ATP synthase in slow- and fast-growing mycobacteria is active in ATP synthesis and blocked in ATP hydrolysis direction.

    NARCIS (Netherlands)

    Haagsma, A.C.; Driessen, N.N.; Hahn, M.M.; Lill, H.; Bald, D.

    2010-01-01

    ATP synthase is a validated drug target for the treatment of tuberculosis, and ATP synthase inhibitors are promising candidate drugs for the treatment of infections caused by other slow-growing mycobacteria, such as Mycobacterium leprae and Mycobacterium ulcerans. ATP synthase is an essential enzyme

  10. ATP synthase in slow- and fast-growing mycobacteria is active in ATP synthesis and blocked in ATP hydrolysis direction.

    NARCIS (Netherlands)

    Haagsma, A.C.; Driessen, N.N.; Hahn, M.M.; Lill, H.; Bald, D.

    2010-01-01

    ATP synthase is a validated drug target for the treatment of tuberculosis, and ATP synthase inhibitors are promising candidate drugs for the treatment of infections caused by other slow-growing mycobacteria, such as Mycobacterium leprae and Mycobacterium ulcerans. ATP synthase is an essential enzyme

  11. How reliable are ATP bioluminescence meters in assessing decontamination of environmental surfaces in healthcare settings?

    Directory of Open Access Journals (Sweden)

    Navid Omidbakhsh

    Full Text Available BACKGROUND: Meters based on adenosine triphosphate (ATP bioluminescence measurements in relative light units (RLU are often used to rapidly assess the level of cleanliness of environmental surfaces in healthcare and other settings. Can such ATP measurements be adversely affected by factors such as soil and cleaner-disinfectant chemistry? OBJECTIVE: This study tested a number of leading ATP meters for their sensitivity, linearity of the measurements, correlation of the readings to the actual microbial contamination, and the potential disinfectant chemicals' interference in their readings. METHODS: First, solutions of pure ATP in various concentrations were used to construct a standard curve and determine linearity and sensitivity. Serial dilutions of a broth culture of Staphylococcus aureus, as a representative nosocomial pathogen, were then used to determine if a given meter's ATP readings correlated with the actual CFUs. Next, various types of disinfectant chemistries were tested for their potential to interfere with the standard ATP readings. RESULTS: All four ATP meters tested herein demonstrated acceptable linearity and repeatability in their readings. However, there were significant differences in their sensitivity to detect the levels of viable microorganisms on experimentally contaminated surfaces. Further, most disinfectant chemistries tested here quenched the ATP readings variably in different ATP meters evaluated. CONCLUSIONS: Apart from their limited sensitivity in detecting low levels of microbial contamination, the ATP meters tested were also prone to interference by different disinfectant chemistries.

  12. Experimental Study of Sorbitol Production by Zymomonas mobilis in High Sucrose Concentration

    OpenAIRE

    Rui Sérgio dos Santos Ferreira da Silva; João Batista Buzato; Maria Antonia Pedrine Colabone Celligoi; Marcos Roberto de Oliveira

    2004-01-01

    The sorbitol presents several industrial applications and its conventional production is of high cost and low yield. Sorbitol production by Zymomonas mobilis production has attracted attention as both production cost and environmental impact are low. The sorbitol plays an osmo-protective rule so that its production is promoted by high sugar concentrations. This work has evaluated the effect of high sucrose concentration in the sorbitol production. The raise of sucrose concentration from 100 t...

  13. The interference of HEPES buffer during amperometric detection of ATP in clinical applications.

    Science.gov (United States)

    Masson, Jean-Francois; Gauda, Estelle; Mizaikoff, Boris; Kranz, Christine

    2008-04-01

    HEPES-based biological buffer is subject to photooxidation upon exposure to fluorescent illumination. Thereby hydrogen peroxide is generated, which interferes with amperometric oxidoreductase-based biosensors for glucose or adenosine triphosphate (ATP). These biosensors operate at an oxidation potential above 500 mV vs. the standard calomel electrode (SCE) and involve hydrogen peroxide as the electroactive molecule detected at the electrode surface. False-positive detection of ATP was observed in HEPES buffer utilizing an amperometric microbiosensor based on the co-immobilization of glucose oxidase and hexokinase for detection of ATP in biological specimens. Electrochemical, mass spectrometric, (31)P NMR, and (1)H NMR studies indicate that complexation of ATP and HEPES induced by the presence of Ca(2+) in HEPES buffer decreases the photooxidation of HEPES. Consequently, the hydrogen peroxide background concentration is reduced, thereby leading to erroneous ATP detection at the dual-enzyme microbiosensor, which determines an increase in ATP via a reduced hydrogen peroxide signal.

  14. Evaluation of ATP bioluminescence assays for potential use in a hospital setting.

    Science.gov (United States)

    Aiken, Zoie A; Wilson, Michael; Pratten, Jonathan

    2011-05-01

    ATP bioluminescence is being applied in hospitals to measure surface contamination. We compared commercial luminometers for detecting the number Staphylococcus aureus associated with surfaces. The data showed that the ATP bioluminescence methods tested were not robust enough to generate quantitative data on bacterial numbers, especially at low concentrations.

  15. An Aptamer-Based Nanobiosensor for Real-Time Measurements of ATP Dynamics

    DEFF Research Database (Denmark)

    Özalp, Cengiz; Nielsen, Lise Junker; Olsen, Lars Folke

    2010-01-01

    A nanosensor based on a new DNA aptamer can show changes in intracellular concentration of ATP of 0.5 to 8 mM and thereby the kinetics of ATP-consuming reactions in real-time. The biosensor was protected against nuclease attack by being buried in a polyacrylamide nanoparticle. This strategy can...

  16. High catechin concentrations detected in Withania somnifera (ashwagandha by high performance liquid chromatography analysis

    Directory of Open Access Journals (Sweden)

    Sulaiman Siti

    2011-08-01

    Full Text Available Abstract Background Withania somnifera is an important medicinal plant traditionally used in the treatment of many diseases. The present study was carried out to characterize the phenolic acids, flavonoids and 1,1-diphenyl-2-picrylhydrazyl radical (DPPH scavenging activities in methanolic extracts of W. somnifera fruits, roots and leaves (WSFEt, WSREt and WSLEt. Methods WSFEt, WSREt and WSLEt was prepared by using 80% aqueous methanol and total polyphenols, flavonoids as well as DPPH radical scavenging activities were determined by spectrophotometric methods and phenolic acid profiles were determined by HPLC methods. Results High concentrations of both phenolics and flavonoids were detected in all parts of the plant with the former ranging between 17.80 ± 5.80 and 32.58 ± 3.16 mg/g (dry weight and the latter ranging between 15.49 ± 1.02 and 31.58 ± 5.07 mg/g. All of the three different plant parts showed strong DPPH radical scavenging activities (59.16 ± 1.20 to 91.84 ± 0.38%. Eight polyphenols (gallic, syringic, benzoic, p-coumaric and vanillic acids as well as catechin, kaempferol and naringenin have been identified by HPLC in parts of the plant as well. Among all the polyphenols, catechin was detected in the highest concentration (13.01 ± 8.93 to 30.61 ± 11.41 mg/g. Conclusion The results indicating that W. somnifera is a plant with strong therapeutic properties thus further supporting its traditional claims. All major parts of W. somnifera such as the roots, fruits and leaves provide potential benefits for human health because of its high content of polyphenols and antioxidant activities with the leaves containing the highest amounts of polyphenols specially catechin with strong antioxidant properties.

  17. ATPase Activity Measurements Using Radiolabeled ATP

    NARCIS (Netherlands)

    Swarts, H.G.; Koenderink, J.B.

    2016-01-01

    ATP provides the energy that is essential for all P-type ATPases to actively transport their substrates against an existing gradient. This ATP hydrolysis can be measured using different methods. Here, we describe a method that uses radiolabeled [gamma-(32)P]ATP, which is hydrolyzed by P-type ATPases

  18. ATP formation and ATP hydrolysis during fatiguing, intermittent stimulation of different types of single muscle fibres from Xenopus laevis.

    Science.gov (United States)

    Nagesser, A S; Van der Laarse, W J; Elzinga, G

    1993-12-01

    This report describes changes of the rate of ATP hydrolysis in single, intact muscle fibres during the development of fatigue induced by intermittent tetanic stimulation. High (type 3) and low (type 1) oxidative muscle fibres dissected from the iliofibularis muscle of Xenopus laevis were studied at 20 degrees C. The rate of ATP hydrolysis was calculated during different time intervals from changes in the content of nucleotides, creatine compounds and lactate, as well as lactate efflux and oxygen uptake. During the first phase of intermittent stimulation, phosphocreatine is fully reduced while the rate of oxygen consumption increases to its maximum, the lactate content increases to a maximum level, and a small amount of IMP is formed; the rate of ATP hydrolysis in type 3 fibres is constant while force decreases, whereas the rate decreases approximately in proportion to force in type 1 fibres. After the first phase, the rate of ATP hydrolysis in type 3 fibres decreases slightly and the fibres reach a steady metabolic state in which the rates of ATP formation and hydrolysis are equal; in type 1 fibres a drastic change of the rate of ATP hydrolysis occurs and a steady metabolic state is not reached. On the basis of the time courses of the metabolic changes, it is concluded that the rate of ATP hydrolysis in type 3 fibres is reduced by acidification and/or a reduced calcium efflux from the sarcoplasmic reticulum, whereas in type 1 fibres inorganic phosphate and/or acidification inhibit the rate initially and ADP is a likely candidate to explain the drastic fall of the rate of ATP hydrolysis during late phases of fatiguing stimulation.

  19. Extracellular ATP does not induce P2X7 receptor-dependent responses in cultured renal- and liver-derived swine macrophages

    Directory of Open Access Journals (Sweden)

    Takato Takenouchi

    2014-01-01

    Full Text Available The P2X7 receptor (P2X7R is an ATP-gated cation channel that is abundantly expressed in monocytes/macrophages. P2X7R activation by ATP results in various cellular responses including Ca2+ influx, membrane pore formation, and cytokine secretion. Since P2X7R has low affinity for ATP, high concentrations of ATP (in the mM range are generally required to activate this receptor in vitro. Functional expression of P2X7R has been detected in monocytes/macrophages obtained from different animal species including humans, rodents, dogs, and bovines, but so far it has not been detected in swine (Sus scrofa. In this study, we investigated the expression and functions of P2X7R in swine macrophages, which were isolated from mixed primary cultures of swine kidney or liver tissue. The P2X7R mRNA and protein expression observed in the swine macrophages was comparable to that seen in a c-myc-immortalized mouse kidney-derived clonal macrophage cell line (KM-1. However, extracellular ATP did not induce P2X7R-dependent sustained Ca2+ influx, membrane pore formation, or the secretion of the bioactive cytokine interleukin-1β in the swine macrophages, whereas these responses were clearly observed in the mouse KM-1 cells after stimulation with millimolar concentrations of ATP as a positive control. These findings suggest that the ATP/P2X7R pathway is impaired in swine macrophages at least in the culture conditions used in the present study.

  20. Metal-dependent regulation of ATP7A and ATP7B in fibroblast cultures

    DEFF Research Database (Denmark)

    Lenartowicz, Malgorzata; Moos, Torben; Ogórek, Mateusz

    2016-01-01

    Deficiency of one of the copper transporters ATP7A and ATP7B leads to the rare X-linked disorder Menkes Disease (MD) or the rare autosomal disorder Wilson disease (WD), respectively. In order to investigate whether the ATP7A and the ATP7B genes may be transcriptionally regulated, we measured...

  1. Metal-Dependent Regulation of ATP7A and ATP7B in Fibroblast Cultures

    DEFF Research Database (Denmark)

    Lenartowicz, Malgorzata; Moos, Torben; Ogórek, Mateusz;

    2016-01-01

    Deficiency of one of the copper transporters ATP7A and ATP7B leads to the rare X-linked disorder Menkes Disease (MD) or the rare autosomal disorder Wilson disease (WD), respectively. In order to investigate whether the ATP7A and the ATP7B genes may be transcriptionally regulated, we measured...

  2. High-Efficiency Solar Thermal Vacuum Demonstration Completed for Refractive Secondary Concentrator

    Science.gov (United States)

    Wong, Wayne A.

    2001-01-01

    Common to many of the space applications that utilize solar thermal energy--such as electric power conversion, thermal propulsion, and furnaces--is a need for highly efficient, solar concentration systems. An effort is underway at the NASA Glenn Research Center to develop the refractive secondary concentrator, which uses refraction and total internal reflection to efficiently concentrate and direct solar energy. When used in combination with advanced lightweight primary concentrators, the refractive secondary concentrator enables very high system concentration ratios (10,000 to 1) and very high temperatures (>2000 K). The innovative refractive secondary concentrator offers significant advantages over all other types of secondary concentrators. The refractive secondary offers the highest throughput efficiency, provides for flux tailoring, requires no active cooling, relaxes the pointing and tracking requirements of the primary concentrator, and enables very high system concentration ratios. This technology has broad applicability to any system that requires the conversion of solar energy to heat. Glenn initiated the development of the refractive secondary concentrator in support of Shooting Star, a solar thermal propulsion flight experiment, and continued the development in support of Space Solar Power.

  3. Phenomenological analysis of ATP dependence of motor proteins.

    Directory of Open Access Journals (Sweden)

    Yunxin Zhang

    Full Text Available In this study, through phenomenological comparison of the velocity-force data of processive motor proteins, including conventional kinesin, cytoplasmic dynein and myosin V, I found that, the ratio between motor velocities of two different ATP concentrations is almost invariant for any substall, superstall or negative external loads. Therefore, the velocity of motors can be well approximated by a Michaelis-Menten like formula V = [ATP]k(FL([ATP] + K(M, with L the step size, and k(F the external load F dependent rate of one mechanochemical cycle of motor motion in saturated ATP solution. The difference of Michaelis-Menten constant K(M for substall, superstall and negative external load indicates, the configurations at which ATP molecule can bind to motor heads for these three cases might be different, though the expression of k(F as a function of F might be unchanged for any external load F. Verifications of this Michaelis-Menten like formula has also been done by fitting to the recent experimental data.

  4. Phenomenological analysis of ATP dependence of motor protein

    CERN Document Server

    Zhang, Yunxin

    2011-01-01

    In this study, through phenomenological comparison of the velocity-force data of processive motor proteins, including conventional kinesin, cytoplasmic dynein and myosin V, we found that, the ratio between motor velocities of two different ATP concentrations is almost invariant for any substall, superstall or negative external loads. Therefore, the velocity of motor can be well approximated by a Michaelis-Menten like formula $V=\\atp k(F)L/(\\atp +K_M)$, with $L$ the step size, and $k(F)$ the external load $F$ dependent rate of one mechanochemical cycle of motor motion in saturated ATP solution. The difference of Michaelis-Menten constant $K_M$ for substall, superstall and negative external load indicates, the ATP molecule affinity of motor head for these three cases are different, though the expression of $k(F)$ as a function of $F$ might be unchanged for any external load $F$. Verifications of this Michaelis-Menten like formula has also been done by fitting to the recent experimental data.

  5. High-sensitivity in situ QCLAS-based ammonia concentration sensor for high-temperature applications

    Science.gov (United States)

    Peng, W. Y.; Sur, R.; Strand, C. L.; Spearrin, R. M.; Jeffries, J. B.; Hanson, R. K.

    2016-07-01

    A novel quantum cascade laser (QCL) absorption sensor is presented for high-sensitivity in situ measurements of ammonia (hbox {NH}_3) in high-temperature environments, using scanned wavelength modulation spectroscopy (WMS) with first-harmonic-normalized second-harmonic detection (scanned WMS-2 f/1 f) to neutralize the effect of non-absorption losses in the harsh environment. The sensor utilized the sQ(9,9) transition of the fundamental symmetric stretch band of hbox {NH}_3 at 10.39 {\\upmu }hbox {m} and was sinusoidally modulated at 10 kHz and scanned across the peak of the absorption feature at 50 Hz, leading to a detection bandwidth of 100 Hz. A novel technique was used to select an optimal WMS modulation depth parameter that reduced the sensor's sensitivity to spectral interference from hbox {H}_2hbox {O} and hbox {CO}_2 without significantly sacrificing signal-to-noise ratio. The sensor performance was validated by measuring known concentrations of hbox {NH}_3 in a flowing gas cell. The sensor was then demonstrated in a laboratory-scale methane-air burner seeded with hbox {NH}_3, achieving a demonstrated detection limit of 2.8 ± 0.26 ppm hbox {NH}_3 by mole at a path length of 179 cm, equivalence ratio of 0.6, pressure of 1 atm, and temperatures of up to 600 K.

  6. Towards a multiscale description of microvascular flow regulation: O2-dependent release of ATP from human erythrocytes and the distribution of ATP in capillary networks

    Directory of Open Access Journals (Sweden)

    Daniel eGoldman

    2012-07-01

    Full Text Available Integration of the numerous mechanisms that have been suggested to contribute to optimization of O2 supply to meet O2 need in skeletal muscle requires a systems biology approach which permits quantification of these physiological processes over a wide range of length scales. Here we describe two individual computational models based on in vivo and in vitro studies which, when incorporated into a single robust multiscale model, will provide information on the role of erythrocyte-released ATP in perfusion distribution in skeletal muscle under both physiological and pathophysiological conditions. Healthy human erythrocytes exposed to low O2 tension release ATP via a well characterized signaling pathway requiring activation of the G-protein, Gi, and adenylyl cyclase leading to increases in cAMP. This cAMP then activates PKA and subsequently CFTR culminating in ATP release via pannexin 1. A critical control point in this pathway is the level of cAMP which is regulated by pathway-specific phosphodiesterases. Using time constants (~100ms that are consistent with measured erythrocyte ATP release, we have constructed a dynamic model of this pathway. The model predicts levels of ATP release consistent with measurements obtained over a wide range of hemoglobin O2 saturations (sO2. The model further predicts how insulin, at concentrations found in prediabetes, enhances the activity of PDE3 and reduces intracellular cAMP levels leading to decreased low O2-induced ATP release from erythrocytes. The second model, which couples O2 and ATP transport in capillary networks, shows how intravascular ATP and the resulting conducted vasodilation are affected by local sO2, convection and ATP degradation. This model also predicts network-level effects of decreased ATP release resulting from elevated insulin levels. Taken together, these models lay the groundwork for investigating the systems biology of the regulation of microvascular perfusion distribution by

  7. Salinity-dependent copper accumulation in the guppy Poecilia vivipara is associated with CTR1 and ATP7B transcriptional regulation.

    Science.gov (United States)

    da Silva, Evelise Sampaio; Abril, Sandra Isabel Moreno; Zanette, Juliano; Bianchini, Adalto

    2014-07-01

    Copper (Cu) accumulation and regulation of key-genes involved in Cu homeostasis were evaluated in freshwater- and saltwater-acclimated guppies Poecilia vivipara. Fish were exposed (96h) to environmentally relevant concentrations of dissolved Cu (0, 5.0, 9.0 and 20.0μg/L). In freshwater guppies, gill and liver Cu accumulation was dependent on Cu concentration in the exposure medium. In saltwater guppies, this dependence was observed only in the gut. These findings indicate that Cu accumulation was salinity- and tissue-dependent. Key genes involved in Cu metabolism were sequenced for the first time in P. vivipara. Transcripts coding for the high-affinity copper transporter (CTR1) and copper-transporting ATPase (ATP7B) were identified using polymerase chain reaction (PCR) and gene sequencing. The full-length CTR1 open reading frame (1560bp) and a partial ATP7B (690bp) were discovered. Predicted amino acid sequences shared high identities with the CTR1 of Fundulus heteroclitus (81%) and the ATP7B of Sparus aurata (87%). Basal transcriptional levels addressed by RT-qPCR in control fish indicate that CTR1 and ATP7B was highly transcribed in liver of freshwater guppies while CTR1 was highly transcribed in gut of saltwater guppies. This could explain the higher Cu accumulation observed in liver of freshwater guppies and in gut of saltwater guppies, because CTR1 is involved in Cu uptake. Reduced gill mRNA expression of CTR1 was observed in freshwater guppies exposed to 20.0μg/L Cu and in saltwater guppies exposed to 5.0μg/L Cu. In turn, reduced mRNA expression of gut ATP7B was observed in freshwater and salt water guppies exposed to 9.0 and 20.0μg/L Cu. Liver CTR1 and ATP7B transcription were not affected by Cu exposure. These findings suggest that gill CTR1 and gut ATP7B are down-regulated to limit Cu absorption after exposure to dissolved Cu, while liver CTR1 and ATP7B levels are maintained to allow Cu storage and detoxification. In conclusion, findings reported here

  8. Dependency of the force-velocity relationships on Mg ATP in different types of muscle fibers from Xenopus laevis.

    OpenAIRE

    Stienen, G J; Laarse, W. J. van der; Elzinga, G.

    1988-01-01

    MgATP binding to the actomyosin complex is followed by the dissociation of actin and myosin. The rate of this dissociation process was determined from the relationship between the maximum velocity of shortening and the MgATP concentration. It is shown here that the overall dissociation rate is rather similar in different types of muscle fibers. The relation between MgATP concentration and the maximum shortening velocity was investigated in fast and slow fibers and bundles of myofibrils of the...

  9. Vitamin C at high concentrations induces cytotoxicity in malignant melanoma but promotes tumor growth at low concentrations.

    Science.gov (United States)

    Yang, Guang; Yan, Yao; Ma, Younan; Yang, Yixin

    2017-08-01

    Vitamin C has been used in complementary and alternative medicine for cancers regardless of its ineffectiveness in clinical trials and the paradoxical effects antioxidants have on cancer. Vitamin C was found to induce cytotoxicity against cancers. However, the mechanisms of action have not been fully elucidated, and the effects of vitamin C on human malignant melanoma have not been examined. This study revealed that vitamin C at millimolar concentrations significantly reduced the cell viability as well as invasiveness, and induced apoptosis in human malignant melanoma cells. Vitamin C displayed stronger cytotoxicity against the Vemurafenib-resistance cell line A2058 compared with SK-MEL-28. In contrast, vitamin C at micromolar concentrations promoted cell growth, migration and cell cycle progression, and protected against mitochondrial stress. Vemurafenib paradoxically activated the RAS-RAF-MEK-ERK signaling pathway in the Vemurafenib-resistant A2058, however, vitamin C abolished the activations. Vitamin C displayed synergistic cytotoxicity with Vemurafenib against the Vemurafenib-resistant A2058. In vivo assay suggested that lower dosage (equivalent to 0.5 g/70 kg) of vitamin C administered orally increased the melanoma growth. Therefore, vitamin C may exert pro- or anti-melanoma effect depending on concentration. The combination of vitamin C at high dosage and Vemurafenib is promising in overcoming the action of drug resistance. © 2017 Wiley Periodicals, Inc.

  10. Localisation de l'ATP synthase mitochondriale et remaniement du réseau mitochondrial en quiescence

    OpenAIRE

    Jimenez, Laure

    2014-01-01

    Mitochondria form a dynamic tubular network which organization and distribution is highly regulated.Mitochondria are double membrane organites with a complex internal architecture. Cristae, which areinner membrane invaginations, are the site of oxidative phosphorylation, reactions by which ATP synthaseproduces ATP. ATP synthase also play a key role in cristae morphogenesis. In this study, I have shown thatATP synthase localized as discrete clusters along the mitochondrial network in living S....

  11. Effect of extraluminal ATP application on vascular tone and blood flow in skeletal muscle

    DEFF Research Database (Denmark)

    Nyberg, Michael Permin; Al-Khazraji, Baraa K; Mortensen, Stefan P

    2013-01-01

    During skeletal muscle contractions, the concentration of ATP increases in muscle interstitial fluid as measured by microdialysis probes. This increase is associated with the magnitude of blood flow, suggesting that interstitial ATP may be important for contraction-induced vasodilation. However......, interstitial ATP has solely been described to induce vasoconstriction in skeletal muscle. To examine whether interstitial ATP induces vasodilation in skeletal muscle and to what extent this vasoactive effect is mediated by formation of nitric oxide (NO) and prostanoids, three different experimental models were...... studied. The rat gluteus maximus skeletal muscle model was used to study changes in local skeletal muscle hemodynamics. Superfused ATP at concentrations found during muscle contractions (1-10 µM) increased blood flow by up to 400%. In this model, the underlying mechanism was also examined by inhibition...

  12. The influence of ferric (III citrate on ATP-hydrolases of Desulfuromonas acetoxidans ІМV В-7384

    Directory of Open Access Journals (Sweden)

    O. Maslovska

    2013-02-01

    Full Text Available Desulfuromonas acetoxidans obtains energy for growth by the anaerobic oxidation of organic compounds with the carbon dioxide formation. It was found that ferrum and manganese are used as terminal electron acceptors in the processes of anaerobic respiration, such as dissimilative Fe3+- and Mn4+-reduction, carried out by these bacteria (Lovely, 1991. D. acetoxidans ІМV B-7384 can be used as anode biocatalyst in microbial fuel cell with high electron recovery through acetate oxidation to the electric current as a result of electron transfer to the anode or 3d-type transition metals, such as ferrum and manganese, in the process of their reduction. Investigation of biochemical changes of D. acetoxidans ІМV B-7384 under the influence of Fe (III compounds is important for optimization of the process of bacterial electricity generation. ATP-hydrolase is located in cytoplasmic membrane, and its subunits are exposed to both the cytoplasm and the external environment. Therefore, the changes of that enzyme activity can be used as an indicator of various stress exposure. Presence of ferric iron ions in the bacterial growth medium could catalyze generation of organic reactive oxygen species, such as peroxyl (ROO- and alkoxyl (RO- radicals. Lipid peroxidation is one of the main reasons of cell damage and it’s following death under the influence of reactive oxygen metabolites. It is known that lipid peroxidation and membrane transport processes are somehow interrelated, but mechanisms of such interaction are still unidentified. In our previous researche we have shown the influence of ferric (III citrate on the intensity of lipid peroxidation of D. аcetoxidans ІМV В-7384. Significant increase of the content of lipid peroxidation products (lipid hydroperoxides, conjugated dienes and malondialdehyde in bacterial cells has been observed under the addition of ferric (III citrate into the cultural medium. The increase of the concentration of lipid

  13. Quantitation of low concentrations of polysorbates in high protein concentration formulations by solid phase extraction and cobalt-thiocyanate derivatization.

    Science.gov (United States)

    Kim, Justin; Qiu, Jinshu

    2014-01-02

    A spectrophotometric method was developed to quantify low polysorbate (PS) levels in biopharmaceutical formulations containing high protein concentrations. In the method, Oasis HLB solid phase extraction (SPE) cartridge was used to extract PS from high protein concentration formulations. After loading a sample, the cartridge was washed with 4M guanidine HCl and 10% (v/v) methanol, and the retained PS was eluted by acetonitrile. Following the evaporation of acetonitrile, aqueous cobalt-thiocyanate reagent was added to react with the polyoxyethylene oxide chain of polysorbates to form a blue colored PS-cobaltothiocyante complex. This colored complex was then extracted into methylene chloride and measured spectrophotometrically at 620 nm. The method performance was evaluated on three products containing 30-40 mg L(-1) PS-20 and PS-80 in ≤70 g L(-1) protein formulations. The method was specific (no matrix interference identified in three types of protein formulations), sensitive (quantitation limit of 10 mg L(-1) PS) and robust with good precision (relative standard deviation ≤6.4%) and accuracy (spike recoveries from 95% to 101%). The linear range of the method for both PS-20 and PS-80 was 10 to 80 mg L(-1) PS. By diluting samples with 6M guanidine HCl and/or using different methylene chloride volumes to extract the colored complexes of standards and samples, the method could accurately and precisely quantify 40 mg L(-1) PS in up to 300 g L(-1) protein formulations.

  14. Induction of Posttranslational Modifications of Mitochondrial Proteins by ATP Contributes to Negative Regulation of Mitochondrial Function.

    Directory of Open Access Journals (Sweden)

    Yong Zhang

    Full Text Available It is generally accepted that ATP regulates mitochondrial function through the AMPK signaling pathway. However, the AMPK-independent pathway remains largely unknown. In this study, we investigated ATP surplus in the negative regulation of mitochondrial function with a focus on pyruvate dehydrogenase (PDH phosphorylation and protein acetylation. PDH phosphorylation was induced by a high fat diet in the liver of obese mice, which was associated with ATP elevation. In 1c1c7 hepatoma cells, the phosphorylation was induced by palmitate treatment through induction of ATP production. The phosphorylation was associated with a reduction in mitochondria oxygen consumption after 4 h treatment. The palmitate effect was blocked by etomoxir, which inhibited ATP production through suppression of fatty acid β-oxidation. The PDH phosphorylation was induced by incubation of mitochondrial lysate with ATP in vitro without altering the expression of PDH kinase 2 (PDK2 and 4 (PDK4. In addition, acetylation of multiple mitochondrial proteins was induced by ATP in the same conditions. Acetyl-CoA exhibited a similar activity to ATP in induction of the phosphorylation and acetylation. These data suggest that ATP elevation may inhibit mitochondrial function through induction of the phosphorylation and acetylation of mitochondrial proteins. The results suggest an AMPK-independent mechanism for ATP regulation of mitochondrial function.

  15. Lipoprotein distribution and serum concentrations of 7α-hydroxy-4-cholesten-3-one and bile acids: effects of monogenic disturbances in high-density lipoprotein metabolism

    DEFF Research Database (Denmark)

    Steiner, Carine; Holleboom, Adriaan G; Karuna, Ratna

    2012-01-01

    A-I (apolipoprotein A-I], ABCA1 (ATP-binding cassette transporter A1) or LCAT (lecithin:cholesterol acyltransferase) nor HDL-C-increasing mutations in the genes CETP (cholesteryl ester transfer protein) or LIPC (hepatic lipase) were associated with significantly different serum concentrations of BA and C4. Plasma...

  16. Sympathetic nerve-derived ATP regulates renal medullary blood flow via vasa recta pericytes

    Directory of Open Access Journals (Sweden)

    Scott S Wildman

    2013-10-01

    Full Text Available Pericyte cells are now known to be a novel locus of blood flow control, being able to regulate capillary diameter via their unique morphology and expression of contractile proteins. We have previously shown that exogenous ATP causes constriction of vasa recta via renal pericytes, acting at a variety of membrane bound P2 receptors on descending vasa recta, and therefore may be able to regulate medullary blood flow (MBF. Regulation of MBF is essential for appropriate urine concentration and providing essential oxygen and nutrients to this region of high, and variable, metabolic demand. Various sources of endogenous ATP have been proposed, including from epithelial, endothelial and red blood cells in response to stimuli such as mechanical stimulation, local acidosis, hypoxia, and exposure to various hormones. Extensive sympathetic innervation of the nephron has previously been shown, however the innervation reported has focused around the proximal and distal tubules, and ascending loop of Henle. We hypothesise that sympathetic nerves are an additional source of ATP acting at renal pericytes and therefore regulate MBF. Using a rat live kidney slice model in combination with video imaging and confocal microscopy techniques we firstly show sympathetic nerves in close proximity to vasa recta pericytes in both the outer and inner medulla. Secondly, we demonstrate pharmacological stimulation of sympathetic nerves in situ (by tyramine evokes pericyte-mediated vasoconstriction of vasa recta capillaries; inhibited by the application of the P2 receptor antagonist suramin. Lastly, tyramine-evoked vasoconstriction of vasa recta by pericytes is significantly less than ATP-evoked vasoconstriction. Sympathetic innervation may provide an additional level of functional regulation in the renal medulla that is highly localized. It now needs to be determined under which physiological/pathophysiological circumstances that sympathetic innervation of renal pericytes is

  17. Antihyperlipidemic activity of adenosine triphosphate in rabbits fed a high-fat diet and hyperlipidemic patients.

    Science.gov (United States)

    Zhang, Lianshan; Liang, Libin; Tong, Tong; Qin, Yuguo; Xu, Yanping; Tong, Xinglong

    2016-10-01

    Context Recently, adenosine triphosphate (ATP) was occasionally found to decrease the triglyceride (TG) levels in several hyperlipidemic patients in our clinical practice. Objective The study investigates the anti-hyperlipidemic effects of ATP in a high-fat fed rabbit model and hyperlipidemic patients. Materials and methods Twenty-four rabbits were randomly divided into three groups of eight animals each as follows: normal diet, high-fat diet and high-fat diet + ATP group. ATP supplementation (40 mg/day) was started at the 20th day and lasted for 10 days. Serum concentrations of total cholesterol (TC), TG, LDL-C, HDL-C were measured on the 20th day and 30th day. Heart, liver and aorta were subjected histopathological examination. Twenty outpatients diagnosed primary hyperlipidemia took ATP at a dose of 60 mg twice a day for 1 week. Results Feeding rabbits with a high-fat diet resulted in a significant elevation of lipid parameters including TC, TG, LDL-C, VLDL-C compared to the normal diet group (p ATP treatment significantly decreased serum TG level (p ATP significantly reduced the thickness of fat layer in cardiac epicardium (p ATP for 1 week, hyperlipidemia patients exhibited a significant decrease of TG (p ATP selectively decreases serum TG levels in high-fat diet rabbits and hyperlipidemic patients. Therefore, ATP supplementation may provide an effective approach to control TG level.

  18. ATP7A (Menkes Protein) functions in Axonal Targeting and Synaptogenesis

    OpenAIRE

    Meskini, Rajaâ El; Crabtree, Kelli L.; Cline, Laura B.; Mains, Richard E.; Eipper, Betty A.; Ronnett, Gabriele V.

    2007-01-01

    Menkes Disease (MD) is a neurodegenerative disorder caused by mutations in the copper transporter, ATP7A, a P-type ATPase. We previously used the olfactory system to demonstrate that ATP7A expression is developmentally, not constitutive, regulated, peaking during synaptogenesis when it is highly expressed in extending axons in a copper-independent manner. Although not known to be associated with axonal functions, we explored the possibility that the inability of mutant ATP7A to support axon o...

  19. Use of ATP to characterize biomass viability in freely suspended and immobilized cell bioreactors

    Energy Technology Data Exchange (ETDEWEB)

    Gikas, P.; Livingston, A.G. (Imperial Coll., London (United Kingdom). Dept. of Chemical Engineering)

    1993-12-01

    This work describes investigations into the viability of cells growing on 3,4-dichloroaniline (34DCA). Two bio-reactors are employed for microbial growth, a continuous stirred tank (CST) bioreactor with a 2-L working volume, and a three-phase air lift (TPAL) bioreactor with a 3-L working volume. Experiments have been performed at several dilution rates between 0.027 and 0.115 h[sup [minus]1] in the CST bioreactor and between 0.111 and 0.500h[sup [minus]1] in the TPAL bioreactor. The specific ATP concentration was calculated at each dilution rate in the suspended biomass in both bioreactors as well as in the immobilized biomass in the TPAL bioreactor. The cultures were inspected under an electron microscope to monitor compositional changes. Results from the CST bioreactor showed that the biomass-specific ATP concentration increases from 0.44 to 1.86 mg ATP g[sup [minus]1] dry weight (dw) as dilution rate increases from 0.027 to 0.115 h[sup [minus]1]. At this upper dilution rate the cells were washed out. The specific ATP concentration reached a limiting average value of 1.73 mg ATP g[sup [minus]1] dw, which is assumed to be the quantity of ATP in 100% viable biomass, In the TPAL bioreactor, the ATP level increased with dilution rat in both the immobilized and suspended biomass. The specific ATP concentration in the immobilized biomass increased from approximately 0.051 mg ATP g[sup [minus]1] dw at dilution rates between 0.111 and 0.200 h[sup [minus]1] to approximately 0.119 mg ATP g[sup [minus]1] dw at dilution rates between 0.300 and 0.500 h[sup [minus]1].

  20. Design, Fabrication and Test of a High Efficiency Refractive Secondary Concentrator for Solar Applications

    Science.gov (United States)

    Wong, Wayne A.; Geng, Steven M.; Castle, Charles H.; Macosko, Robert P.

    2000-01-01

    Common to many of the space applications that utilize solar thermal energy such as electric power conversion, thermal propulsion, and furnaces, is a need for highly efficient, solar concentration systems. An effort is underway to develop the refractive secondary concentrator, which uses refraction and total internal reflection to efficiently concentrate and direct solar energy. When used in combination with advanced primary concentrators, the refractive secondary concentrator enables very high system concentration ratios (10,000 to 1) and very high temperatures (greater than 2000 K). Presented is an overview of the effort at the NASA Glenn Research Center to evaluate the performance of a prototype single crystal sapphire refractive secondary concentrator and to compare the performance with analytical models. The effort involves the design and fabrication of a secondary concentrator, design and fabrication of a calorimeter and its support hardware, calibration of the calorimeter, testing of the secondary concentrator in NASA Glenn's Tank 6 solar thermal vacuum facility, and comparing the test results with predictions. Test results indicate an average throughput efficiency of 87%. It is anticipated that reduction of a known reflection loss with an anti-reflective coating would result in a secondary concentrator throughput efficiency of approximately 93%.

  1. ATP sensitizes H460 lung carcinoma cells to cisplatin-induced apoptosis.

    Science.gov (United States)

    Swennen, Els L R; Ummels, Vanessa; Buss, Irina; Jaehde, Ulrich; Bast, Aalt; Dagnelie, Pieter C

    2010-03-30

    Platinum resistance of cancer cells may evolve due to a decrease in intracellular drug accumulation, decreased cell permeability or by an increased deactivation of the drug by glutathione (GSH). The aim of this study was (1) to investigate the effect of adenosine 5'-triphosphate (ATP) on the cytotoxicity of cisplatin in a large cell lung carcinoma cell line (H460), and (2) to examine the potential involvement of increased cisplatin uptake, GSH depletion and pyrimidine starvation by ATP in this effect. H460 cells were harvested and seeded (5% CO(2); 37 degrees C). Subsequently, cells were incubated with medium or ATP followed by an incubation with cisplatin. Cytotoxicity screening was analyzed by the sulforhodamine B (SRB) colorimetric assay, lactate dehydrogenase and caspase-3/7 activity. Pre-incubation for 72h with 0.3 and 3mM ATP strongly enhanced the anti-proliferative potency of cisplatin 2.9- and 7.6-fold, respectively. Moreover, after incubation of H460 cells with 0.3mM ATP the intracellular platinum concentration increased, indicating increased cisplatin uptake by ATP. ATP, despite lowering the LD(50) of cisplatin, did not modulate GSH levels in H460 cells. ATP itself showed a biphasic effect on H460 cell growth: 0.3mM inhibited H460 cell growth via the pyrimidine starvation effect, activation of caspase-3/7 and LDH leakage, while 3mM ATP showed no effect on cell growth. In conclusion, ATP sensitizes the H460 cells to cisplatin-induced apoptosis. The effect of 0.3mM ATP is not due to GSH depletion but involves increased cisplatin uptake and pyrimidine starvation due to ATP conversion to adenosine followed by cellular uptake.

  2. Combining biofunctional magnetic nanoparticles and ATP bioluminescence for rapid detection of Escherichia coli.

    Science.gov (United States)

    Cheng, Yuxiao; Liu, Yajun; Huang, Jingjing; Li, Kang; Zhang, Wen; Xian, Yuezhong; Jin, Litong

    2009-02-15

    A rapid, specific and sensitive method for assay of Escherichia coli (E. coli) using biofunctional magnetic nanoparticles (BMNPs) in combination with adenosine triphosphate (ATP) bioluminescence was proposed. The BMNPs were fabricated by immobilizing a specific anti-E. coli antibody on the surface of amine-functionalized magnetic nanoparticles (about 20nm in diameter), and then was applied to capture the target bacteria E. coli from samples. The BMNPs exhibited high capture efficiency to E. coli. Transmission electron microscope (TEM) images showed that the BMNPs were bound to the surface of entire E. coli cells. The target bacteria became magnetic so that could be isolated easily from the sample solution by employing an external magnetic field. The concentration of E. coli captured by the BMNPs was then detected by an ATP bioluminescence method. The optimization of ATP measurement was carried out to improve the detection sensitivity. The proposed method was applied to detect the E. coli inoculated into pasteurized milk with low detection limit (20 cfu/mL) and short detection time (about 1h).

  3. Urinary Hypoxanthine as a Measure of Increased ATP Utilization in Late Preterm Infants

    Science.gov (United States)

    Holden, Megan S.; Hopper, Andrew; Slater, Laurel; Asmerom, Yayesh; Esiaba, Ijeoma; Boskovic, Danilo S.; Angeles, Danilyn M.

    2015-01-01

    Objective To examine the effect of neonatal morbidity on ATP breakdown in late preterm infants. Study Design Urinary hypoxanthine concentration, a marker of ATP breakdown, was measured from 82 late preterm infants on days of life (DOL) 3 to 6 using high-performance liquid chromatography. Infants were grouped according to the following diagnoses: poor nippling alone (n = 8), poor nippling plus hyperbilirubinemia (n = 21), poor nippling plus early respiratory disease (n = 26), and respiratory disease alone (n = 27). Results Neonates with respiratory disease alone had significantly higher urinary hypoxanthine over DOL 3 to 6 when compared with neonates with poor nippling (P = .020), poor nippling plus hyperbilirubinemia (P < .001), and poor nippling plus early respiratory disease (P = .017). Neonates with poor nippling who received respiratory support for 2 to 3 days had significantly higher hypoxanthine compared with infants who received respiratory support for 1 day (P = .017) or no days (P = .007). Conclusions These findings suggest that respiratory disorders significantly increase ATP degradation in late premature infants. PMID:26413195

  4. Identification of small molecules that improve ATP synthesis defects conferred by Leber's hereditary optic neuropathy mutations.

    Science.gov (United States)

    Datta, Sandipan; Tomilov, Alexey; Cortopassi, Gino

    2016-09-01

    Inherited mitochondrial complex I mutations cause blinding Leber's hereditary optic neuropathy (LHON), for which no curative therapy exists. A specific biochemical consequence of LHON mutations in the presence of trace rotenone was observed: deficient complex I-dependent ATP synthesis (CIDAS) and mitochondrial O2 consumption, proportional to the clinical severity of the three primary LHON mutations. We optimized a high-throughput assay of CIDAS to screen 1600 drugs to 2, papaverine and zolpidem, which protected CIDAS in LHON cells concentration-dependently. TSPO and cAMP were investigated as protective mechanisms, but a conclusive mechanism remains to be elucidated; next steps include testing in animal models.

  5. Novel DDR Processing of Corn Stover Achieves High Monomeric Sugar Concentrations from Enzymatic Hydrolysis (230 g/L) and High Ethanol Concentration (10% v/v) During Fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Xiaowen; Jennings, Ed; Shekiro, Joe; Kuhn, Erik M.; O' Brien, Marykate; Wang, Wei; Schell, Daniel J.; Himmel, Mike; Elander, Richard T.; Tucker, Melvin P.

    2015-04-03

    Distilling and purifying ethanol, butanol, and other products from second and later generation lignocellulosic biorefineries adds significant capital and operating cost for biofuels production. The energy costs associated with distillation affects plant gate and life cycle analysis costs. Lower titers in fermentation due to lower sugar concentrations from pretreatment increase both energy and production costs. In addition, higher titers decrease the volumes required for enzymatic hydrolysis and fermentation vessels. Therefore, increasing biofuels titers has been a research focus in renewable biofuels production for several decades. In this work, we achieved over 200 g/L of monomeric sugars after high solids enzymatic hydrolysis using the novel deacetylation and disc refining (DDR) process on corn stover. The high sugar concentrations and low chemical inhibitor concentrations from the DDR process allowed ethanol titers as high as 82 g/L in 22 hours, which translates into approximately 10 vol% ethanol. To our knowledge, this is the first time that 10 vol% ethanol in fermentation derived from corn stover without any sugar concentration or purification steps has been reported. Techno-economic analysis shows the higher titer ethanol achieved from the DDR process could significantly reduce the minimum ethanol selling price from cellulosic biomass.

  6. Thermodynamically consistent description of the hydrodynamics of free surfaces covered by insoluble surfactants of high concentration

    OpenAIRE

    Thiele, Uwe; Archer, Andrew J.; Plapp, Mathis

    2012-01-01

    In this paper we propose several models that describe the dynamics of liquid films which are covered by a high concentration layer of insoluble surfactant. First, we briefly review the 'classical' hydrodynamic form of the coupled evolution equations for the film height and surfactant concentration that are well established for small concentrations. Then we re-formulate the basic model as a gradient dynamics based on an underlying free energy functional that accounts for wettability and capill...

  7. Incomplete Ion Dissociation Underlies the Weakened Attraction between DNA Helices at High Spermidine Concentrations

    OpenAIRE

    Yang,Jie; Rau, Donald C

    2005-01-01

    We have investigated the salt sensitivity of the hexagonal-to-cholesteric phase transition of spermidine-condensed DNA. This transition precedes the resolubilization of precipitated DNA that occurs at high spermidine concentration. The sensitivity of the critical spermidine concentration at the transition point to the anion species and the NaCl concentration indicates that ion pairing of this trivalent ion underlies this unusual transition. Osmotic pressure measurements of spermidine salt sol...

  8. Atovaquone and quinine anti-malarials inhibit ATP binding cassette transporter activity

    NARCIS (Netherlands)

    Rijpma, S.R.; Heuvel, J.J.; Velden, M. van der; Sauerwein, R.W.; Russel, F.G.; Koenderink, J.B.

    2014-01-01

    BACKGROUND: Therapeutic blood plasma concentrations of anti-malarial drugs are essential for successful treatment. Pharmacokinetics of pharmaceutical compounds are dependent of adsorption, distribution, metabolism, and excretion. ATP binding cassette (ABC) transport proteins are particularly involve

  9. Effect of digestion time on anaerobic digestion with high ammonia concentration

    Science.gov (United States)

    Oktavitri, Nur Indradewi; Purnobasuki, Hery; Kuncoro, Eko Prasetyo; Purnamasari, Indah; Semma Hadinnata, P.

    2016-03-01

    Anaerobic digestion was developed to treat high concentration organic compound efficiently in certain Digestion Time (DT). High ammonia concentration could influenced removal organic compound in digestion. This bench scale study investigated the effect of digestion time on anaerobic batch reactor with high ammonia concentration. Total Ammonia Nitrogen (TAN) concentration was adjusted 4000 and 5000 mg/1, Digestion time was ranged from 0-26 d, operation temperature was ranged from 28-29°C, inoculum was collected from slaughter house sludge. The degradation of Chemical Oxygen Demand (COD) correlated with digestion time. The concentration of TAN from synthetic wastewater contain 5000 mg/1 of TAN more fluctuated than those use 4000 mg/1 of TAN. However, the biogas production from wastewater contained 4000 mg/1 of TAN gradually increased until 24 d of DT. The reactor contain 5000 mg/1 of TAN only growth until 12 d and steady state at over 12 d of digestion time.

  10. Structure-guided simulations illuminate the mechanism of ATP transport through VDAC1.

    Science.gov (United States)

    Choudhary, Om P; Paz, Aviv; Adelman, Joshua L; Colletier, Jacques-Philippe; Abramson, Jeff; Grabe, Michael

    2014-07-01

    The voltage-dependent anion channel (VDAC) mediates the flow of metabolites and ions across the outer mitochondrial membrane of all eukaryotic cells. The open channel passes millions of ATP molecules per second, whereas the closed state exhibits no detectable ATP flux. High-resolution structures of VDAC1 revealed a 19-stranded β-barrel with an α-helix partially occupying the central pore. To understand ATP permeation through VDAC, we solved the crystal structure of mouse VDAC1 (mVDAC1) in the presence of ATP, revealing a low-affinity binding site. Guided by these coordinates, we initiated hundreds of molecular dynamics simulations to construct a Markov state model of ATP permeation. These simulations indicate that ATP flows through VDAC through multiple pathways, in agreement with our structural data and experimentally determined physiological rates.

  11. Melatonin in Arabidopsis thaliana acts as plant growth regulator at low concentrations and preserves seed viability at high concentrations.

    Science.gov (United States)

    Hernández, Ismaél Gatica; Gomez, Federico José Vicente; Cerutti, Soledad; Arana, María Verónica; Silva, María Fernanda

    2015-09-01

    Since the discovery of melatonin in plants, several roles have been described for different species, organs, and developmental stages. Arabidopsis thaliana, being a model plant species, is adequate to contribute to the elucidation of the role of melatonin in plants. In this work, melatonin was monitored daily by UHPLC-MS/MS in leaves, in order to study its diurnal accumulation as well as the effects of natural and artificial light treatments on its concentration. Furthermore, the effects of exogenous application of melatonin to assess its role in seed viability after heat stress and as a regulator of growth and development of vegetative tissues were evaluated. Our results indicate that melatonin contents in Arabidopsis were higher in plants growing under natural radiation when compared to those growing under artificial conditions, and its levels were not diurnally-regulated. Exogenous melatonin applications prolonged seed viability after heat stress conditions. In addition, melatonin applications retarded leaf senescence. Its effects as growth promoter were dose and tissue-dependent; stimulating root growth at low concentrations and decreasing leaf area at high doses.

  12. Prodigious Effects of Concentration Intensification on Nanoparticle Synthesis: A High-Quality, Scalable Approach.

    Science.gov (United States)

    Williamson, Curtis B; Nevers, Douglas R; Hanrath, Tobias; Robinson, Richard D

    2015-12-23

    Realizing the promise of nanoparticle-based technologies demands more efficient, robust synthesis methods (i.e., process intensification) that consistently produce large quantities of high-quality nanoparticles (NPs). We explored NP synthesis via the heat-up method in a regime of previously unexplored high concentrations near the solubility limit of the precursors. We discovered that in this highly concentrated and viscous regime the NP synthesis parameters are less sensitive to experimental variability and thereby provide a robust, scalable, and size-focusing NP synthesis. Specifically, we synthesize high-quality metal sulfide NPs (200 g) relative to the current field of large-scale (0.1-5 g yields) and laboratory-scale (<0.1 g) efforts. Compared to conventional synthesis methods (hot injection with dilute precursor concentration) characterized by rapid growth and low yield, our highly concentrated NP system supplies remarkably controlled growth rates and a 10-fold increase in NP volumetric production capacity (86 g/L). The controlled growth, high yield, and robust nature of highly concentrated solutions can facilitate large-scale nanomanufacturing of NPs by relaxing the synthesis requirements to achieve monodisperse products. Mechanistically, our investigation of the thermal and rheological properties and growth rates reveals that this high concentration regime has reduced mass diffusion (a 5-fold increase in solution viscosity), is stable to thermal perturbations (∼64% increase in heat capacity), and is resistant to Ostwald ripening.

  13. High-concentration-gradient dispersion in porous media : experiments, analysis and approximations

    NARCIS (Netherlands)

    R.J. Schotting; H. Moser; S.M. Hassanizadeh

    1997-01-01

    textabstractVarious experimental and theoretical studies have shown that Fick's law, based on the assumption of a linear relation between solute dispersive mass flux and concentration gradient, is not valid when high concentration gradients are encountered in a porous medium. The value of the macrod

  14. Critical phenomena in ethylbenzene oxidation in acetic acid solution at high cobalt(II) concentrations

    NARCIS (Netherlands)

    Gavrichkov, AA; Zakharov, [No Value

    2005-01-01

    Critical phenomena in ethylbenzene oxidation in an acetic acid solution at high cobalt(ill) concentrations (from 0.01 to 0.2 mol L-1) were studied at 60-90 degrees C by the gasometric (O-2 absorption), spectrophotometric (Co-III accumulation), and chemiluminescence (relative concentration of radical

  15. Whey protein particles modulate mechanical properties of gels at high protein concentrations

    NARCIS (Netherlands)

    Saglam, D.; Venema, P.; Vries, de R.J.; Berg, van den L.; Linden, van der E.

    2014-01-01

    We have studied the influence of dense whey protein particles on the mechanical properties of whey protein isolate (WPI) gels at high protein concentrations (16–22% (w/w)). Incorporation of dense whey protein particles in the gel, while keeping the total protein concentration constant, leads to a co

  16. Rheology of dilute acid hydrolyzed corn stover at high solids concentration

    Science.gov (United States)

    M.R. Ehrhardt; T.O. Monz; T.W. Root; R.K. Connelly; Tim Scott; D.J. Klingenberg

    2010-01-01

    The rheological properties of acid hydrolyzed corn stover at high solids concentration (20–35 wt.%) were investigated using torque rheometry. These materials are yield stress fluids whose rheological properties can be well represented by the Bingham model. Yield stresses increase with increasing solids concentration and decrease with increasing hydrolysis reaction...

  17. Effect of dissolved oxygen concentration on the bioflocculation process in high loaded MBRs

    NARCIS (Netherlands)

    Faust, L.; Temmink, B.G.; Zwijnenburg, A.; Kemperman, A.J.B.; Rijnaarts, H.

    2014-01-01

    High-loaded membrane bioreactors (HL-MBRs), i.e. MBRs which are operated at extremely short sludge and hydraulic retention times, can be applied to flocculate and concentrate sewage organic matter. The concentrated organics can be used for energy recovery, or for the production of more valuable

  18. ATP6V0C knockdown in neuroblastoma cells alters autophagy-lysosome pathway function and metabolism of proteins that accumulate in neurodegenerative disease.

    Directory of Open Access Journals (Sweden)

    Leandra R Mangieri

    Full Text Available ATP6V0C is the bafilomycin A1-binding subunit of vacuolar ATPase, an enzyme complex that critically regulates vesicular acidification. We and others have shown previously that bafilomycin A1 regulates cell viability, autophagic flux and metabolism of proteins that accumulate in neurodegenerative disease. To determine the importance of ATP6V0C for autophagy-lysosome pathway function, SH-SY5Y human neuroblastoma cells differentiated to a neuronal phenotype were nucleofected with non-target or ATP6V0C siRNA and following recovery were treated with either vehicle or bafilomycin A1 (0.3-100 nM for 48 h. ATP6V0C knockdown was validated by quantitative RT-PCR and by a significant decrease in Lysostracker Red staining. ATP6V0C knockdown significantly increased basal levels of microtubule-associated protein light chain 3-II (LC3-II, α-synuclein high molecular weight species and APP C-terminal fragments, and inhibited autophagic flux. Enhanced LC3 and LAMP-1 co-localization following knockdown suggests that autophagic flux was inhibited in part due to lysosomal degradation and not by a block in vesicular fusion. Knockdown of ATP6V0C also sensitized cells to the accumulation of autophagy substrates and a reduction in neurite length following treatment with 1 nM bafilomycin A1, a concentration that did not produce such alterations in non-target control cells. Reduced neurite length and the percentage of propidium iodide-positive dead cells were also significantly greater following treatment with 3 nM bafilomycin A1. Together these results indicate a role for ATP6V0C in maintaining constitutive and stress-induced ALP function, in particular the metabolism of substrates that accumulate in age-related neurodegenerative disease and may contribute to disease pathogenesis.

  19. Rotation and structure of FoF1-ATP synthase.

    Science.gov (United States)

    Okuno, Daichi; Iino, Ryota; Noji, Hiroyuki

    2011-06-01

    F(o)F(1)-ATP synthase is one of the most ubiquitous enzymes; it is found widely in the biological world, including the plasma membrane of bacteria, inner membrane of mitochondria and thylakoid membrane of chloroplasts. However, this enzyme has a unique mechanism of action: it is composed of two mechanical rotary motors, each driven by ATP hydrolysis or proton flux down the membrane potential of protons. The two molecular motors interconvert the chemical energy of ATP hydrolysis and proton electrochemical potential via the mechanical rotation of the rotary shaft. This unique energy transmission mechanism is not found in other biological systems. Although there are other similar man-made systems like hydroelectric generators, F(o)F(1)-ATP synthase operates on the nanometre scale and works with extremely high efficiency. Therefore, this enzyme has attracted significant attention in a wide variety of fields from bioenergetics and biophysics to chemistry, physics and nanoscience. This review summarizes the latest findings about the two motors of F(o)F(1)-ATP synthase as well as a brief historical background.

  20. A High-Efficiency Refractive Secondary Solar Concentrator for High Temperature Solar Thermal Applications

    Science.gov (United States)

    Piszczor, Michael F., Jr.; Macosko, Robert P.

    2000-01-01

    A refractive secondary solar concentrator is a non-imaging optical device that accepts focused solar energy from a primary concentrator and redirects that light, by means of refraction and total internal reflection (TIR) into a cavity where the solar energy is used for power and/or propulsion applications. This concept offers a variety of advantages compared to typical reflective secondary concentrators (or the use of no secondary at all): higher optical efficiency, minimal secondary cooling requirements, a smaller cavity aperture, a reduction of outgassing from the cavity and flux tailoring of the solar energy within the heat receiver. During the past 2 years, NASA Lewis has been aggressively developing this concept in support of the NASA Marshall Shooting Star Flight Experiment. This paper provides a brief overview of the advantages and technical challenges associated with the development of a refractive secondary concentrator and the fabrication of a working unit in support of the flight demonstration program.

  1. Stimulation-dependent release, breakdown, and action of endogenous ATP in mouse hemidiaphragm preparation: the possible role of ATP in neuromuscular transmission.

    Science.gov (United States)

    Vizi, E S; Nitahara, K; Sato, K; Sperlágh, B

    2000-07-01

    In this study the in vitro mouse phrenic nerve- hemidiaphragm preparation was utilized to study the release and extracellular catabolism of endogenous ATP and its action on the postsynaptic site, i.e. on the contraction force evoked by nerve stimulation. ATP, measured by the luciferin-luciferase assay, was released stimulation-dependently from the mouse hemidiaphragm in response to electrical field stimulation at 10 Hz. Blockade of the Na(+) channel activity by tetrodotoxin inhibited the majority of the release of ATP in response to stimulation, showing that it is related to neuronal activity. The nicotinic receptor antagonists d-tubocurarine, and alpha-bungarotoxin and cooling the bath temperature to 7 degrees C also reduced stimulation-induced ATP outflow, suggesting that nicotinic receptors are responsible for the part of the release of ATP that is released from postsynaptic sites in a carrier-mediated manner. Exogenous ATP (20-500 microM) added to the bath was degraded to ADP and AMP by the action of ectoATPase and ectoATPdiphosphohydrolase; the K(m) and v(max) values of these enzymes were 185.8 microM and 55.16 nmol/min.g respectively. However, the total amount of nucleotides ([ATP+ADP+AMP]) was increased after the addition of ATP, indicating that ATP itself promoted further adenine nucleotide release. Twitch contractions of the rat hemidiaphragm preparation evoked by low frequency electrical stimulation was blocked concentration-dependently by the non-depolarizing muscle relaxants d-tubocurarine and pancuronium. Suramin (100 microM-1 mM) reversed neuromuscular blockade by d-tubocurarine and pancuronium; i.e., it shifted their concentration-response curves to the right Taken together our data, that endogenous ATP is released by stimulation and subsequently catabolized in the hemidiaphragm preparation and that suramin inhibits ecto-ATPase activity could be interpreted as meaning that suramin prolongs the action of endogenous ATP to elicit twitch contraction

  2. Ultrasensitive bioluminescent determinations of adenosine triphosphate (ATP) for investigating the energetics of host-grown microbes

    Science.gov (United States)

    Hanks, J. H.; Dhople, A. M.

    1975-01-01

    Stability and optimal concentrations of reagents were studied in bioluminescence assay of ATP levels. Luciferase enzyme was prepared and purified using Sephadex G-100. Interdependencies between enzyme and luciferin concentrations in presence of optimal Mg are illustrated. Optimal ionic strength was confirmed to be 0.05 M for the four buffers tested. Adapted features of the R- and H-systems are summarized, as well as the percentages of ATP pools released from representative microbes by heat and chloroform.

  3. ATP enhances spontaneous calcium activity in cultured suburothelial myofibroblasts of the human bladder.

    Directory of Open Access Journals (Sweden)

    Sheng Cheng

    Full Text Available BACKGROUND: Suburothelial myofibroblasts (sMF are located underneath the urothelium in close proximity to afferent nerves. They express purinergic receptors and show calcium transients in response to ATP. Therefore they are supposed to be involved in afferent signaling of the bladder fullness. Since ATP concentration is likely to be very low during the initial filling phase, we hypothesized that sMF Ca(2+ activity is affected even at very low ATP concentrations. We investigated ATP induced modulation of spontaneous activity, intracellular calcium response and purinergic signaling in cultured sMF. METHODOLOGY/PRINCIPAL FINDINGS: Myofibroblast cultures, established from cystectomies, were challenged by exogenous ATP in presence or absence of purinergic antagonist. Fura-2 calcium imaging was used to monitor ATP (10(-16 to 10(-4 mol/l induced alterations of calcium activity. Purinergic receptors (P2X1, P2X2, P2X3 were analysed by confocal immunofluorescence. We found spontaneous calcium activity in 55.18% ± 1.65 of the sMF (N = 48 experiments. ATP significantly increased calcium activity even at 10(-16 mol/l. The calcium transients were partially attenuated by subtype selective antagonist (TNP-ATP, 1 µM; A-317491, 1 µM, and were mimicked by the P2X1, P2X3 selective agonist α,β-methylene ATP. The expression of purinergic receptor subtypes in sMF was confirmed by immunofluorescence. CONCLUSIONS/SIGNIFICANCE: Our experiments demonstrate for the first time that ATP can modulate spontaneous activity and induce intracellular Ca(2+ response in cultured sMF at very low concentrations, most likely involving P2X receptors. These findings support the notion that sMF are able to register bladder fullness very sensitively, which predestines them for the modulation of the afferent bladder signaling in normal and pathological conditions.

  4. Investigating cold gelation properties of recombined highly concentrated micellar casein concentrate and cream for use in cheese making.

    Science.gov (United States)

    Lu, Y; McMahon, D J; Vollmer, A H

    2016-07-01

    Highly concentrated micellar casein concentrate (HC-MCC), a potential ingredient for cheese making, contains ~20% casein with ~70% of serum proteins removed by microfiltration and diafiltration of skim milk, followed by vacuum evaporation. Our objective was to investigate cold gelation properties of recombined concentrated milk (RCM) by mixing thawed frozen HC-MCC and cream under different casein levels, pH, and protein-to-fat ratios, and with addition of sodium citrate or calcium. The HC-MCC was recombined with cream using low shear at 50°C for 30 min, and rheological measurements were conducted. Cold-gelling temperature [the temperature at which storage modulus (G')=loss modulus (G″)] was linearly correlated with casein levels from 8.6 to 11.5% (R(2)=0.71), pH from 6.6 to 7.0 (R(2)=0.96), and addition of sodium citrate from 0 to 0.36mmol/g of casein (R(2)=0.80). At pH 7.0, gelation occurred at 12, 26, and 38°C with 9, 10, and 11% casein, respectively. At pH 6.6, 6.8, and 7.0, RCM with 12% casein gelled at a mean temperature of 12, 26, and 37°C, respectively. Adding calcium chloride at 0.17mmol/g of casein significantly increased cold-gelling temperature from 18 to ≥50°C, whereas no significant change was observed at levels up to 0.12mmol/g of casein. Different protein to fat ratios ranging from 0.8 to 1.2 did not significantly influence gelling temperature. In transmission electron micrographs of RCM with 12% casein, casein micelles were nonspherical and partially dissociated into small protein strands. Upon addition of calcium chloride at 0.21mmol/g of casein, casein micelles were more spherical and retained colloidal structure with the presence of aggregated casein micelles. These gelation processes of RCM with or without addition of trisodium citrate were both reversible. We propose that cold gelation of RCM occurs when protein strands that have been partially released from the casein micelles entangle, restrict their mobility, and form a fine

  5. Visualization and measurement of ATP levels in living cells replicating hepatitis C virus genome RNA.

    Directory of Open Access Journals (Sweden)

    Tomomi Ando

    Full Text Available Adenosine 5'-triphosphate (ATP is the primary energy currency of all living organisms and participates in a variety of cellular processes. Although ATP requirements during viral lifecycles have been examined in a number of studies, a method by which ATP production can be monitored in real-time, and by which ATP can be quantified in individual cells and subcellular compartments, is lacking, thereby hindering studies aimed at elucidating the precise mechanisms by which viral replication energized by ATP is controlled. In this study, we investigated the fluctuation and distribution of ATP in cells during RNA replication of the hepatitis C virus (HCV, a member of the Flaviviridae family. We demonstrated that cells involved in viral RNA replication actively consumed ATP, thereby reducing cytoplasmic ATP levels. Subsequently, a method to measure ATP levels at putative subcellular sites of HCV RNA replication in living cells was developed by introducing a recently-established Förster resonance energy transfer (FRET-based ATP indicator, called ATeam, into the NS5A coding region of the HCV replicon. Using this method, we were able to observe the formation of ATP-enriched dot-like structures, which co-localize with non-structural viral proteins, within the cytoplasm of HCV-replicating cells but not in non-replicating cells. The obtained FRET signals allowed us to estimate ATP concentrations within HCV replicating cells as ∼5 mM at possible replicating sites and ∼1 mM at peripheral sites that did not appear to be involved in HCV replication. In contrast, cytoplasmic ATP levels in non-replicating Huh-7 cells were estimated as ∼2 mM. To our knowledge, this is the first study to demonstrate changes in ATP concentration within cells during replication of the HCV genome and increased ATP levels at distinct sites within replicating cells. ATeam may be a powerful tool for the study of energy metabolism during replication of the viral genome.

  6. Extracellular ATP induces spikes in cytosolic free Ca2+ but not in NADH oxidase activity in neutrophils

    DEFF Research Database (Denmark)

    Brasen, Jens Christian; Olsen, Lars Folke; Hallett, Maurice B.

    2011-01-01

    In order to establish whether non-mitochondrial oxidase activity in human neutrophils is tightly related to cytosolic Ca2+ concentration, we simultaneously measured Ca2+ oscillations induced by ATP and oxidant production in single adherent neutrophils using confocal microscopy. ATP induced fast...... that the generation of reactive oxygen species by neutrophils adherent to glass was accelerated by ATP. The step-up in NADPH oxidase activity followed the first elevation of cytosolic Ca2+ but, despite subsequent spikes in Ca2+ concentration, no oscillations in oxidase activity could be detected. ATP induced spikes...

  7. Potassium double tungstate waveguides with high ytterbium concentration for optical amplification

    NARCIS (Netherlands)

    Yong, Yean-Sheng

    2017-01-01

    In this thesis, the research work concerning high ytterbium concentration potassium double tungstate waveguides catered for optical amplification purpose is presented. The scope of the research work includes the investigation of spectroscopic and optical gain properties in epitaxy layers with

  8. Methods to assess high-resolution subsurface gas concentrations and gas fluxes in wetland ecosystems

    DEFF Research Database (Denmark)

    Elberling, Bo; Kühl, Michael; Glud, Ronnie N.

    2013-01-01

    The need for measurements of soil gas concentrations and surface fluxes of greenhouse gases at high temporal and spatial resolution in wetland ecosystem has lead to the introduction of several new analytical techniques and methods. In addition to the automated flux chamber methodology for high......-resolution estimates of greenhouse gas fluxes across the soil-atmosphere interface, these high-resolution methods include microsensors for quantification of spatiotemporal concentration dynamics in O2 and N2O at micrometer scales, fiber-optic optodes for long-term continuous point measurements of O2 concentrations...... and peat soils are highly heterogeneous, containing a mosaic of dynamic macropore systems created by both macrofauna and flora leading to distinct spatial and temporal variations in gas concentration on a scale of millimeters and minutes. Applications of these new methodologies allow measurements...

  9. Concentration influences on recovery in a high gradient magnetic separation axial filter

    Energy Technology Data Exchange (ETDEWEB)

    Murariu, V.; Rezlescu, N.; Rotariu, O.; Badescu, V. [Inst. of Technical Physics, Iasi (Romania)

    1998-05-01

    The buildup differential equations for the case of a single wire in high gradient magnetic filtration (HGMF)-axial configuration taking into account the suspension concentration are solved. A new equation for the deposit contour surface at different moments and for different suspension concentrations are obtained. The existence of a particulate suspension concentration, for which the radial extension velocity of deposit is maximum, is evidenced. The recovery for an ordered ferromagnetic matrix is calculated. The influence of the solid particle concentration from suspension on the filtration efficiency is presented.

  10. Optimisation of ATP determination in drinking water

    DEFF Research Database (Denmark)

    Corfitzen, Charlotte B.; Albrechtsen, Hans-Jørgen

    Adenosine Triphosphate (ATP) can be used as a relative measure of cell activity, and is measured by the light output from the reaction between luciferin and ATP catalyzed by firefly luciferase. The measurement has potential as a monitoring and surveillance tool within drinking water distribution......, since the method is very sensitive (detects 0.5 ng ATP/L) and results are obtained within minutes. When calculating the ATP value a number of parameters need to be considered. These were investigate by use of two different reagent kits (PCP-kit and Lumin(ATE)/Lumin(EX)-kit), internal standard...... and an Advance Coupe luminometer. The investigations showed a 60 times higher response of the PCP-kit, making it more suitable for measurement of samples with low ATP content. ATP-standard dilutions prepared in tap water were stable for at least 15 months when stored frozen at -80ºC, and storage of large...

  11. Optimisation of ATP determination in drinking water

    DEFF Research Database (Denmark)

    Corfitzen, Charlotte B.; Albrechtsen, Hans-Jørgen

    Adenosine Triphosphate (ATP) can be used as a relative measure of cell activity, and is measured by the light output from the reaction between luciferin and ATP catalyzed by firefly luciferase. The measurement has potential as a monitoring and surveillance tool within drinking water distribution......, since the method is very sensitive (detects 0.5 ng ATP/L) and results are obtained within minutes. When calculating the ATP value a number of parameters need to be considered. These were investigate by use of two different reagent kits (PCP-kit and Lumin(ATE)/Lumin(EX)-kit), internal standard...... and an Advance Coupe luminometer. The investigations showed a 60 times higher response of the PCP-kit, making it more suitable for measurement of samples with low ATP content. ATP-standard dilutions prepared in tap water were stable for at least 15 months when stored frozen at -80ºC, and storage of large...

  12. Subcutaneous absorption kinetics of two highly concentrated preparations of recombinant human growth hormone

    DEFF Research Database (Denmark)

    Laursen, Torben; Jørgensen, Jens Otto Lunde; Susgaard, Søren;

    1993-01-01

    Abstract OBJECTIVE: The relative bioavailability of two highly concentrated (12 IU/ml) formulations of biosynthetic human growth hormone (GH) administered subcutaneously was compared. DESIGN: A randomized, crossover study. Conventional GH therapy was withdrawn 72 hours before each study period. T....... CONCLUSIONS: There is no significant difference between the absorption kinetics and short-term metabolic effects of these two highly concentrated formulations of biosynthetic GH. The two formulations are bioequivalent....

  13. Incomplete ion dissociation underlies the weakened attraction between DNA helices at high spermidine concentrations.

    Science.gov (United States)

    Yang, Jie; Rau, Donald C

    2005-09-01

    We have investigated the salt sensitivity of the hexagonal-to-cholesteric phase transition of spermidine-condensed DNA. This transition precedes the resolubilization of precipitated DNA that occurs at high spermidine concentration. The sensitivity of the critical spermidine concentration at the transition point to the anion species and the NaCl concentration indicates that ion pairing of this trivalent ion underlies this unusual transition. Osmotic pressure measurements of spermidine salt solutions are consistent with this interpretation. Spermidine salts are not fully dissociated at higher concentrations. The competition for DNA binding among the fully charged trivalent ion and the lesser charged complex species at higher concentrations significantly weakens attraction between DNA helices in the condensed state. This is contrary to the suggestion that the binding of spermidine at higher concentrations causes DNA overcharging and consequent electrostatic repulsion.

  14. Transcriptional analysis of adaptation to high glucose concentrations in Zymomonas mobilis.

    Science.gov (United States)

    Zhang, Kun; Shao, Huanhuan; Cao, Qinghua; He, Ming-Xiong; Wu, Bo; Feng, Hong

    2015-02-01

    The ethanologenic bacterium Zymomonas mobilis is usually tolerant to high concentrations of glucose. The addition of sorbitol decreases the lag phase and increases ethanol yield and productivity of the bacteria in high glucose concentrations. The molecular mechanisms of adaptation to high glucose concentrations and the effect of sorbitol are still unclear. In this study, microarray analysis was used to study the global transcriptional adaptation responses of Z. mobilis to high glucose concentrations. A total of 235 genes were differentially expressed when 220 g/L glucose was added with or without 10 mM sorbitol. These genes are involved in diverse aspects of cell metabolism and regulation, including membrane transporters, nitrogen metabolism, and plasmid-encoded genes. However, most differentially expressed genes were downregulated when sorbitol was added. Notably, the transcription of almost all genes involved in the Entner-Doudoroff and ethanol production pathways was not significantly affected. In addition, a prophage and a nitrogen-fixation cluster were significantly induced. These results revealed that Z. mobilis cells responded to high glucose concentrations by regulating the transcriptional levels of genes related to membrane channels and transporters, stress response mechanisms, and metabolic pathways. These data provide insight into the intracellular adaptation responses to high glucose concentrations and reveal strategies to engineer efficient ethanol fermentation in Z. mobilis.

  15. Functional studies of ATP sulfurylase from Penicillium chrysogenum

    Energy Technology Data Exchange (ETDEWEB)

    Seubert, P.A.

    1985-01-01

    ATP sulfurylase from Penicillium chrysogenum has a specific activity (V/sub max/) of 6-7 units x mg protein/sup -1/ determined with the physiological substrates of MgATP and SO/sub 4//sup 2 -/ and assayed by (A) initial velocity measurements with APS kinase and inorganic pyrophosphatase present and (B) analysis of nonlinear reaction progress curves. The fact both assays give the same results show the intrinsic activity of ATP sulfurylase is much higher than previously reported. In initial velocity dead-end inhibition studies, the sulfate analog S/sub 2/O/sub 3//sup 2 -/ is a competitive inhibitor of SO/sub 42/..sqrt.. and a noncompetitive inhibitor of MgATP. Monovalent oxyanions such as NO/sub 3//sup -/, ClO/sub 3//sup -/, ClO/sub 4//sup -/, and FSO/sub 3//sup -/ behave as uncompetitive inhibitors of MgATP and thus seem not to be true sulfate analogs. The reverse reaction was assayed by the pyrophosphate dependent release of /sup 35/SO/sub 4//sup 2 -/ from AP/sup 35/S. Product inhibition by MgATP or SO/sub 4//sup 2 -/ is competitive with APS and mixed-type with PP/sub i/. Imidodiphosphate can serve as an alternative substrate for PP/sub i/. ATP sulfurylase binds (but does not hydrolyze) APS. A Scatchard plot of the APS binding is nonlinear, suggesting at least two types of sites. The cumulative results are qualitatively consistent with the random addition of MgATP and SO/sub 4//sup 2 -/ and the ordered release of first MgPP/sub i/ then APS, with APS release being partially rate limiting. Certain quantitative discrepancies suggest either an unknown variable (e.g. enzyme concentration) complicates the analysis or, in light of binding studies that the actual mechanism is more complicated (e.g. alternating sites) than any of the conventional models examined.

  16. EXPERIMENTAL INVESTIGATIONS ON DIFFUSION CHARACTERISTICS OF HIGH CONCENTRATION JETS IN ENVIRONMENTAL CURRENTS

    Institute of Scientific and Technical Information of China (English)

    张燕; 王道增; 樊靖郁

    2002-01-01

    By means of flow visualization and quantitative measurement, the diffusionpattern and concentration distribution characteristics of high concentration jets verticallydischarged into shallow moving waterbody were experimentally investigated in waterchannel. The interactions between the high concentration jets and environmental flowconditions were analysed, and the formulae of impinging point coordinate and transversespread angle are gained from data analysis. Experimental results indicate that the jets showcomplicated flow patterns and diffusion characteristics in near region, which are differentfrom common submerged jets, and spread downstream in the manner of density currents.

  17. Substantia nigra osmoregulation: taurine and ATP involvement.

    Science.gov (United States)

    Morales, Ingrid; Dopico, Jose G; Sabate, Magdalena; Gonzalez-Hernandez, Tomas; Rodriguez, Manuel

    2007-05-01

    An extracellular nonsynaptic taurine pool of glial origin was recently reported in the substantia nigra (SN). There is previous evidence showing taurine as an inhibitory neurotransmitter in the SN, but the physiological role of this nonsynaptic pool of taurine has not been explored. By using microdialysis methods, we studied the action of local osmolarity on the nonsynaptic taurine pool in the SN of the rat. Hypoosmolar pulses (285-80 mosM) administered in the SN by the microdialysis probe increased extrasynaptic taurine in a dose-dependent way, a response that was counteracted by compensating osmolarity with choline. The opposite effect (taurine decrease) was observed when osmolarity was increased. Under basal conditions, the blockade of either the AMPA-kainate glutamate receptors with 6-cyano-7-nitroquinoxaline-2,3-dionine disodium or the purinergic receptors with pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid modified the taurine concentration, suggesting that both receptors modulate the extrasynaptic pool of taurine. In addition, these drugs decreased the taurine response to hypoosmolar pulses, suggesting roles for glutamatergic and purinergic receptors in the taurine response to osmolarity. The participation of purinergic receptors was also supported by the fact that ATP (which, under basal conditions, increased the extrasynaptic taurine in a dose-dependent way) administered in doses saturating purinergic receptors also decreased the taurine response to hypoosmolarity. Taken together, present data suggest osmoregulation as a role of the nonsynaptic taurine pool of the SN, a function that also involves glutamate and ATP and that could influence the nigral cell vulnerability in Parkinson's disease.

  18. A luciferase based viability assay for ATP detection in 384-well format for high throughput whole cell screening of Trypanosoma brucei brucei bloodstream form strain 427

    Directory of Open Access Journals (Sweden)

    Avery Vicky M

    2009-11-01

    Full Text Available Abstract Background Human African Trypanosomiasis (HAT is caused by two trypanosome species, Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense. Current drugs available for the treatment of HAT have significant issues related to toxicity, administration regimes with limited effectiveness across species and disease stages, thus there is a considerable need to find alternative drugs. A well recognised approach to identify new drug candidates is high throughput screening (HTS of large compound library collections. Results We describe here the development of a luciferase based viability assay in 384-well plate format suitable for HTS of T.b.brucei. The parameters that were explored to determine the final HTS assay conditions are described in detail and include DMSO tolerability, Z', diluents and cell inoculum density. Reference compound activities were determined for diminazene, staurosporine and pentamidine and compared to previously published IC50 data obtained. The assay has a comparable sensitivity to reference drugs and is more cost effective than the 96-well format currently reported for T.b.brucei. Conclusion Due to the reproducibility and sensitivity of this assay it is recommended for potential HTS application. As it is commercially available this assay can also be utilised in many laboratories for both large and small scale screening.

  19. Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in Syntrophus aciditrophicus, a New Twist on ATP Formation

    Directory of Open Access Journals (Sweden)

    Kimberly L. James

    2016-08-01

    Full Text Available Syntrophus aciditrophicus is a model syntrophic bacterium that degrades key intermediates in anaerobic decomposition, such as benzoate, cyclohexane-1-carboxylate, and certain fatty acids, to acetate when grown with hydrogen-/formate-consuming microorganisms. ATP formation coupled to acetate production is the main source for energy conservation by S. aciditrophicus. However, the absence of homologs for phosphate acetyltransferase and acetate kinase in the genome of S. aciditrophicus leaves it unclear as to how ATP is formed, as most fermentative bacteria rely on these two enzymes to synthesize ATP from acetyl coenzyme A (CoA and phosphate. Here, we combine transcriptomic, proteomic, metabolite, and enzymatic approaches to show that S. aciditrophicus uses AMP-forming, acetyl-CoA synthetase (Acs1 for ATP synthesis from acetyl-CoA. acs1 mRNA and Acs1 were abundant in transcriptomes and proteomes, respectively, of S. aciditrophicus grown in pure culture and coculture. Cell extracts of S. aciditrophicus had low or undetectable acetate kinase and phosphate acetyltransferase activities but had high acetyl-CoA synthetase activity under all growth conditions tested. Both Acs1 purified from S. aciditrophicus and recombinantly produced Acs1 catalyzed ATP and acetate formation from acetyl-CoA, AMP, and pyrophosphate. High pyrophosphate levels and a high AMP-to-ATP ratio (5.9 ± 1.4 in S. aciditrophicus cells support the operation of Acs1 in the acetate-forming direction. Thus, S. aciditrophicus has a unique approach to conserve energy involving pyrophosphate, AMP, acetyl-CoA, and an AMP-forming, acetyl-CoA synthetase.

  20. Sensitivity of P-glycoprotein tryptophan residues to benzodiazepines and ATP interaction.

    Science.gov (United States)

    Lima, Sofia A C; Cordeiro-da-Silva, Anabela; de Castro, Baltazar; Gameiro, Paula

    2007-01-01

    Plasma membrane P-glycoprotein is a member of the ATP-binding cassette family of membrane transporters. In the present study tryptophan intrinsic fluorescence was used to understand the P-glycoprotein response to three benzodiazepines (bromazepam, chlordiazepoxide and flurazepam) in the presence and absence of ATP. Fluorescence emission spectra showed a red shift on the maximal emission wavelength upon interaction of P-glycoprotein with all benzodiazepines. Benzodiazepine association with nucleotide-bound P-glycoprotein also showed this trend and the quenching profile was attributed to a sphere-of-action model, for static fluorescence. Furthermore, quenching data of benzodiazepine-bound P-glycoprotein with ATP were concentration dependent and saturable, indicating that nucleotide binds to P-glycoprotein whether drug is present or not. These results seems in agreement with the proposal of the ATP-switch model by Higgins and Linton, where substrate binding to the transporters initiates the transport cycle by increasing the ATP binding affinity.

  1. [Study of high temperature water vapor concentration measurement method based on absorption spectroscopy].

    Science.gov (United States)

    Chen, Jiu-ying; Liu, Jian-guo; He, Jun-feng; He, Ya-bai; Zhang, Guang-le; Xu, Zhen-yu; Gang, Qiang; Wang, Liao; Yao, Lu; Yuan, Song; Ruan, Jun; Dai, Yun-hai; Kan, Rui-feng

    2014-12-01

    Tunable diode laser absorption spectroscopy (TDLAS) has been developed to realize the real-time and dynamic measurement of the combustion temperature, gas component concentration, velocity and other flow parameters, owing to its high sensitivity, fast time response, non-invasive character and robust nature. In order to obtain accurate water vapor concentration at high temperature, several absorption spectra of water vapor near 1.39 μm from 773 to 1273 K under ordinary pressure were recorded in a high temperature experiment setup using a narrow band diode laser. The absorbance of high temperature absorption spectra was calculated by combined multi-line nonlinear least squares fitting method. Two water vapor absorption lines near 7154.35 and 7157.73 cm(-1) were selected for measurement of water vapor at high temperature. A model method for high temperature water vapor concentration was first proposed. Water vapor concentration from the model method at high temperature is in accordance with theoretical reasoning, concentration measurement standard error is less than 0.2%, and the relative error is less than 6%. The feasibility of this measuring method is verified by experiment.

  2. Teacher Development Program for ATP 2000. Project Report.

    Science.gov (United States)

    Sutphin, Dean; And Others

    Agri Tech Prep 2000 (ATP 2000) is a 4-year tech prep program linking high school and postsecondary curricula designed to prepare New York students for careers in agriculture or acceptance into a college program in agriculture. Because teacher development was designated an integral project component for fiscal year 1991-1992, a weeklong teacher…

  3. Trace element concentrations and gastrointestinal parasites of Arctic terns breeding in the Canadian High Arctic.

    Science.gov (United States)

    Provencher, J F; Braune, B M; Gilchrist, H G; Forbes, M R; Mallory, M L

    2014-04-01

    Baseline data on trace element concentrations are lacking for many species of Arctic marine birds. We measured essential and non-essential element concentrations in Arctic tern (Sterna paradisaea) liver tissue and brain tissue (mercury only) from Canada's High Arctic, and recorded the presence/absence of gastrointestinal parasites during four different phases of the breeding season. Arctic terns from northern Canada had similar trace element concentrations to other seabird species feeding at the same trophic level in the same region. Concentrations of bismuth, selenium, lead and mercury in Arctic terns were high compared to published threshold values for birds. Selenium and mercury concentrations were also higher in Arctic terns from northern Canada than bird species sampled in other Arctic areas. Selenium, mercury and arsenic concentrations varied across the time periods examined, suggesting potential regional differences in the exposure of biota to these elements. For unknown reasons, selenium concentrations were significantly higher in birds with gastrointestinal parasites as compared to those without parasites, while bismuth concentrations were higher in Arctic terns not infected with gastrointestinal parasites.

  4. High serum uric acid concentration predicts poor survival in patients with breast cancer.

    Science.gov (United States)

    Yue, Cai-Feng; Feng, Pin-Ning; Yao, Zhen-Rong; Yu, Xue-Gao; Lin, Wen-Bin; Qian, Yuan-Min; Guo, Yun-Miao; Li, Lai-Sheng; Liu, Min

    2017-08-26

    Uric acid is a product of purine metabolism. Recently, uric acid has gained much attraction in cancer. In this study, we aim to investigate the clinicopathological and prognostic significance of serum uric acid concentration in breast cancer patients. A total of 443 female patients with histopathologically diagnosed breast cancer were included. After a mean follow-up time of 56months, survival was analysed using the Kaplan-Meier method. To further evaluate the prognostic significance of uric acid concentrations, univariate and multivariate Cox regression analyses were applied. Of the clinicopathological parameters, uric acid concentration was associated with age, body mass index, ER status and PR status. Univariate analysis identified that patients with increased uric acid concentration had a significantly inferior overall survival (HR 2.13, 95% CI 1.15-3.94, p=0.016). In multivariate analysis, we found that high uric acid concentration is an independent prognostic factor predicting death, but insufficient to predict local relapse or distant metastasis. Kaplan-Meier analysis indicated that high uric acid concentration is related to the poor overall survival (p=0.013). High uric acid concentration predicts poor survival in patients with breast cancer, and might serve as a potential marker for appropriate management of breast cancer patients. Copyright © 2017. Published by Elsevier B.V.

  5. Alternative mitochondrial functions in cell physiopathology: beyond ATP production

    Directory of Open Access Journals (Sweden)

    Kowaltowski A.J.

    2000-01-01

    Full Text Available It is well known that mitochondria are the main site for ATP generation within most tissues. However, mitochondria also participate in a surprising number of alternative activities, including intracellular Ca2+ regulation, thermogenesis and the control of apoptosis. In addition, mitochondria are the main cellular generators of reactive oxygen species, and may trigger necrotic cell death under conditions of oxidative stress. This review concentrates on these alternative mitochondrial functions, and their role in cell physiopathology.

  6. Vacquinol-1 inducible cell death in glioblastoma multiforme is counter regulated by TRPM7 activity induced by exogenous ATP.

    Science.gov (United States)

    Sander, Philip; Mostafa, Haouraa; Soboh, Ayman; Schneider, Julian M; Pala, Andrej; Baron, Ann-Kathrin; Moepps, Barbara; Wirtz, C Rainer; Georgieff, Michael; Schneider, Marion

    2017-05-23

    Glioblastomas (GBM) are the most malignant brain tumors in humans and have a very poor prognosis. New therapeutic options are urgently needed. A novel drug, Vacquinol-1 (Vac), a quinolone derivative, displays promising properties by inducing rapid cell death in GBM but not in non-transformed tissues. Features of this type of cell death are compatible with a process termed methuosis. Here we tested Vac on a highly malignant glioma cell line observed by long-term video microscopy. Human dental-pulp stem cells (DPSCs) served as controls. A major finding was that an exogenous ATP concentration of as little as 1 μM counter regulated the Vac-induced cell death. Studies using carvacrol, an inhibitor of transient receptor potential cation channel, subfamily M, member 7 (TRPM7), demonstrated that the ATP-inducible inhibitory effect is likely to be via TRPM7. Exogenous ATP is of relevance in GBM with large necrotic areas. Our results support the use of GBM cultures with different grades of malignancy to address their sensitivity to methuosis. The video-microscopy approach presented here allows decoding of signaling pathways as well as mechanisms of chemotherapeutic resistance by long-term observation. Before implementing Vac as a novel therapeutic drug in GBM, cells from each individual patient need to be assessed for their ATP sensitivity. In summary, the current investigation supports the concept of methuosis, described as non-apoptotic cell death and a promising approach for GBM treatment. Tissue-resident ATP/necrosis may interfere with this cell-death pathway but can be overcome by a natural compound, carvacrol that even penetrates the blood-brain barrier.

  7. An incidence of very high phosphate concentrations in the waters around Andaman Islands

    Digital Repository Service at National Institute of Oceanography (India)

    Reddy, C.V.G.; Murty, P.S.N.; Sankaranarayanan, V.N.

    at stations close to coasts; 2) vertical distribution of phosphates in 200 metre column does not appear to correspond to any regular accepted pattern and 3) all the phosphates having the abnormally high phosphate concentration were found to be highly turbid...

  8. Influence of Mechanical Activation on Acid Leaching Dephosphorization of High-phosphorus Iron Ore Concentrates

    Institute of Scientific and Technical Information of China (English)

    De-qing ZHU; Hao WANG; Jian PAN; Cong-cong YANG

    2016-01-01

    High pressure roll grinding (HPRG)and ball milling were compared to investigate the influence of me-chanical activation on the acid leaching dephosphorization of a high-phosphorus iron ore concentrate,which was man-ufactured through magnetizing roasting-magnetic separation of high-phosphorus oolitic iron ores.The results indica-ted that when high-phosphorus iron ore concentrates containing 54·92 mass% iron and 0·76 mass% phosphorus were directly processed through acid leaching,iron ore concentrates containing 55·74 mass% iron and 0·33 mass%phosphorus with an iron recovery of 84·64% and dephosphorization of 63·79% were obtained.When high-phosphor-us iron ore concentrates activated by ball milling were processed by acid leaching,iron ore concentrates containing 56·03 mass% iron and 0·21 mass% phosphorus with an iron recovery of 85·65% and dephosphorization of 77·49%were obtained.Meanwhile,when high-phosphorus iron ore concentrates activated by HPRG were processed by acid leaching,iron ore concentrates containing 58·02 mass% iron and 0·10 mass% phosphorus were obtained,with the iron recovery reaching 88·42% and the dephosphorization rate reaching 88·99%.Mechanistic studies demonstrated that ball milling can reduce the particle size,demonstrating a prominent reunion phenomenon.In contrast,HPRG pretreatment contributes to the formation of more cracks within the particles and selective dissociation of iron and P bearing minerals,which can provide the favorable kinetic conditions to accelerate the solid-liquid reaction rate.As such,the crystal structure is destroyed and the surface energy of mineral particles is strengthened by mechanical ac-tivation,further strengthening the dephosphorization.

  9. Prodigious Effects of Concentration Intensification on Nanoparticle Synthesis: A High-Quality, Scalable Approach

    KAUST Repository

    Williamson, Curtis B.

    2015-12-23

    © 2015 American Chemical Society. Realizing the promise of nanoparticle-based technologies demands more efficient, robust synthesis methods (i.e., process intensification) that consistently produce large quantities of high-quality nanoparticles (NPs). We explored NP synthesis via the heat-up method in a regime of previously unexplored high concentrations near the solubility limit of the precursors. We discovered that in this highly concentrated and viscous regime the NP synthesis parameters are less sensitive to experimental variability and thereby provide a robust, scalable, and size-focusing NP synthesis. Specifically, we synthesize high-quality metal sulfide NPs (<7% relative standard deviation for Cu2-xS and CdS), and demonstrate a 10-1000-fold increase in Cu2-xS NP production (>200 g) relative to the current field of large-scale (0.1-5 g yields) and laboratory-scale (<0.1 g) efforts. Compared to conventional synthesis methods (hot injection with dilute precursor concentration) characterized by rapid growth and low yield, our highly concentrated NP system supplies remarkably controlled growth rates and a 10-fold increase in NP volumetric production capacity (86 g/L). The controlled growth, high yield, and robust nature of highly concentrated solutions can facilitate large-scale nanomanufacturing of NPs by relaxing the synthesis requirements to achieve monodisperse products. Mechanistically, our investigation of the thermal and rheological properties and growth rates reveals that this high concentration regime has reduced mass diffusion (a 5-fold increase in solution viscosity), is stable to thermal perturbations (64% increase in heat capacity), and is resistant to Ostwald ripening.

  10. ATP - bioluminescence as a technique to evaluate the microbiological quality of water in food industry

    Directory of Open Access Journals (Sweden)

    Patrícia Dolabela Costa

    2004-07-01

    Full Text Available ATP-bioluminescence was used to evaluate the microbiological quality of water samples collected from the water supply, the water treatment system and from a dairy plant, including ammonia-cooling water and industrial water. For industrial water, there was relation between the ATP-bioluminescence technique and microbial count. There were no differences (p>0.05 between water supply and ammonia-cooling water samples for total and free ATP concentrations nor for the microbial counts. Different microbial ATP concentrations were found for these water samples. The results suggested that the physical chemical quality of ammonia cooling water decreased the RLU measurements slightly. It could be concluded that the total ATP concentration was the most effective technique to evaluate the microbiological quality of water used in the food indsutry by ATP-bioluminescence.A qualidade microbiológica da água do manancial de captação para tratamento na ETA/UFV, da água de resfriamento de amônia e da água industrial de uso em um laticínio foi avaliada pela técnica de ATP-bioluminescência. Nas águas, foram efetuadas, também, as contagens de mesófilos aeróbios, expressos em UFC.mL-1 e coliformes totais, expressos em NMP.100 mL-1. Foi utilizado um luminômetro para os testes de determinação de ATP total e livre, expressos em Unidades Relativas de Luz (URL, nas diversas águas. O ATP microbiano foi determinado pela diferença entre o ATP total e livre. Em relação à água industrial, houve a concordância entre os métodos de bioluminescência e de contagem de mesófilos aeróbios e coliformes totais. As amostras de água de manancial e de resfriamento não apresentaram diferença (p>0,05, pelo teste de Tukey, para as quantidades de ATP total, livre e também para as contagens microbianas. Constataram-se concentrações diferentes de ATP microbiano para essas amostras de água. Os resultados indicam que o teste de ATP total é o mais recomendado para

  11. Design and Evaluation of the Highly Concentrated Human IgG Formulation Using Cyclodextrin Polypseudorotaxane Hydrogels.

    Science.gov (United States)

    Higashi, Taishi; Tajima, Anna; Ohshita, Naoko; Hirotsu, Tatsunori; Abu Hashim, Irhan Ibrahim; Motoyama, Keiichi; Koyama, Sawako; Iibuchi, Ruriko; Mieda, Shiuhei; Handa, Kenji; Kimoto, Tomoaki; Arima, Hidetoshi

    2015-12-01

    To achieve the potent therapeutic effects of human immunoglobulin G (IgG), highly concentrated formulations are required. However, the stabilization for highly concentrated human IgG is laborious work. In the present study, to investigate the potentials of polypseudorotaxane (PPRX) hydrogels consisting of polyethylene glycol (PEG) and α- or γ-cyclodextrin (α- or γ-CyD) as pharmaceutical materials for highly concentrated human IgG, we designed the PPRX hydrogels including human IgG and evaluated their pharmaceutical properties. The α- and γ-CyDs formed PPRX hydrogels with PEG (M.W. 20,000) even in the presence of highly concentrated human IgG (>100 mg/mL). According to the results of (1)H-NMR, powder X-ray diffraction, and Raman microscopy, the formation of human IgG/CyD PPRX hydrogels was based on physical cross-linking arising from their columnar structures. The release profiles of human IgG from the hydrogels were in accordance with the non-Fickian diffusion model. Importantly, the stabilities of human IgG included into the hydrogels against thermal and shaking stresses were markedly improved. These findings suggest that PEG/CyD PPRX hydrogels are useful to prepare the formulation for highly concentrated human IgG.

  12. High Concentrating GaAs Cell Operation Using Optical Waveguide Solar Energy System

    Science.gov (United States)

    Nakamura, T.; Case, J. A.; Timmons, M. L.

    2004-01-01

    This paper discusses the result of the concentrating photovoltaic (CPV) cell experiments conducted with the Optical Waveguide (OW) Solar Energy System. The high concentration GaAs cells developed by Research Triangle Institute (RTI) were combined with the OW system in a "fiber-on-cell" configuration. The sell performance was tested up to the solar concentration of 327. Detailed V-I characteristics, power density and efficiency data were collected. It was shown that the CPV cells combined with the OW solar energy system will be an effective electric power generation device.

  13. Photorespiration and carbon concentrating mechanisms: two adaptations to high O2, low CO2 conditions.

    Science.gov (United States)

    Moroney, James V; Jungnick, Nadine; Dimario, Robert J; Longstreth, David J

    2013-11-01

    This review presents an overview of the two ways that cyanobacteria, algae, and plants have adapted to high O2 and low CO2 concentrations in the environment. First, the process of photorespiration enables photosynthetic organisms to recycle phosphoglycolate formed by the oxygenase reaction catalyzed by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Second, there are a number of carbon concentrating mechanisms that increase the CO2 concentration around Rubisco which increases the carboxylase reaction enhancing CO2 fixation. This review also presents possibilities for the beneficial modification of these processes with the goal of improving future crop yields.

  14. High Voltage Solar Concentrator Experiment with Implications for Future Space Missions - S6a-35

    Science.gov (United States)

    George, I. S. Mehdi P. J.; O'Neill, M.; Matson, R.; Borckschmidt, A.

    2004-12-01

    This paper describes the design, development, fabrication, and test of a high performance, high voltage solar concentrator array. This assembly is believed to be the first ever terrestrial triple-junction-cell solar array rated at over 1 kW. The concentrator provides over 200 W/square meter power output at a nominal 600 Vdc while operating under terrestrial sunlight. Space-quality materials and fabrication techniques were used for the array, and the 3005-meter elevation installation below the Tropic of Cancer allowed testing as close as possible to space deployment without an actual launch. The array includes two concentrator modules, each with a 3 square meter aperture area. Each concentrator module uses a linear Fresnel lens to focus sunlight onto a photovoltaic receiver that uses 240 series-connected triple-junction solar cells. Operation of the two receivers in series can provide 1200 Vdc which would be adequate for the "direct drive" of some ion engines or microwave transmitters in space. Lens aperture width is 84 cm and the cell active width is 3.2 cm, corresponding to a geometric concentration ratio of 26X. The evaluation includes the concentrator modules, the solar cells, and the materials and techniques used to attach the solar cells to the receiver heat sink. For terrestrial applications, a finned aluminum extrusion was used for the heat sink for the solar cells, maintaining a low cell temperature so that solar cell efficiency remains high.

  15. Application of Hydrothermal Treatment to High Concentrated Sewage Sludge for Anaerobic Digestion Process

    Directory of Open Access Journals (Sweden)

    M. Orikawa

    2013-10-01

    Full Text Available Tomato and seaweed were produced by utilizing CO2 and heat discharged from power generation using biogas in Toyogawa biomass park, Japan. The biogas was obtained by anaerobic digestion with hydrothermal treatment. The hydrothermal treatment was applied to the high concentrated sewage sludge (22 % total solids (TS dewatered sludge. The purpose of this study is to clarify the effect of hydrothermal treatment on the qualities of high concentrated sewage sludge, by analyzing particulate organic carbon (POC and dissolved organic carbon (DOC. The hydrothermal treatment was investigated under 10-60 min of treatment time, 180-200 °C of temperature, 10-22 %-TS of sewage sludge concentration. The results showed that the DOC in each conditions increased through hydrothermal treatment. The highest DOC obtained was 67 % of total carbon concentration, when the temperature was 180 °C, treatment time was 60 min and sewage sludge concentration was 10 %-TS. Furthermore, the viscosity of treated sewage sludge was decreased by hydrothermal treatment. In batch anaerobic digestion test, methane gas production was confirmed. In addition, this study evaluated the energy balance of this system. Thus, the results of this study indicated that the possibility of application of hydrothermal treatment to high concentrated sewage sludge for anaerobic digestion process. Keywords: anaerobic reaction, hydrothermal treatment, sewage sludge, solubilization

  16. High concentrations of myeloperoxidase in the equine uterus as an indicator of endometritis.

    Science.gov (United States)

    Parrilla-Hernandez, Sonia; Ponthier, Jérôme; Franck, Thierry Y; Serteyn, Didier D; Deleuze, Stéfan C

    2014-04-15

    Intraluminal fluid and excessive abnormal hyperedema are regularly used for the diagnosis of endometritis in the mare, which is routinely confirmed by the presence of neutrophils on endometrial smears. Studies show a relation between neutrophils and myeloperoxidase (MPO), an enzyme contained in and released by neutrophils during degranulation or after cell lysis. This enzyme has been found in many fluids and tissues, and associated with different inflammatory pathologies in the horse. The aims of this study were to assess the presence and concentration of MPO in the equine uterus, and to investigate its relation with neutrophils, and other clinical signs of endometritis. Mares (n = 51) were evaluated for the presence of intraluminal fluid and excessive endometrial edema before breeding, and a small volume lavage and cytology samples were obtained. From 69 cycles, supernatant of the uterine flushes was analyzed with a specific equine MPO ELISA assay to measure MPO concentration. Cytology samples were used for the diagnosis of endometritis. Myeloperoxidase was present in the uterus of all estrus mares in highly variable concentrations. Myeloperoxidase concentrations were significantly (P < 0.05) higher in samples with positive cytologies and in the presence of intraluminal fluid. Occasionally, some samples with negative cytologies showed high MPO concentration, but the opposite was never observed. Cycles presenting hyperedema weren't associated with high concentration of MPO, intraluminal fluid, or positive cytology, making it a poor diagnostic tool of endometritis.

  17. ATP and Presentation Service for Mizar Formalizations

    CERN Document Server

    Urban, Josef; Sitcliffe, Geoff

    2011-01-01

    This paper describes the Automated Reasoning for Mizar (MizAR) service, which integrates several automated reasoning, artificial intelligence, and presentation tools with Mizar and its authoring environment. The service provides ATP assistance to Mizar authors in finding and explaining proofs, and offers generation of Mizar problems as challenges to ATP systems. The service is based on a sound translation from the Mizar language to that of first-order ATP systems, and relies on the recent progress in application of ATP systems in large theories containing tens of thousands of available facts. We present the main features of MizAR services, followed by an account of initial experiments in finding proofs with the ATP assistance. Our initial experience indicates that the tool offers substantial help in exploring the Mizar library and in preparing new Mizar articles.

  18. The reaction process of firefly bioluminescence triggered by photolysis of caged-ATP.

    Science.gov (United States)

    Kageyama, Takeshi; Tanaka, Masatoshi; Sekiya, Takao; Ohno, Shin-Ya; Wada, Naohisa

    2011-01-01

    The reaction process of firefly bioluminescence was studied by photolyzing caged-ATP to adenosine triphosphate (ATP) within 100 ms. The intensity of luminescence increases markedly to reach a maximum within 1 s, maintains almost the same intensity up to 5 s and then decays monotonically. The rise γ(1) and decay γ(2) rate constants were determined to be about 5 s(-1) and 1 × 10(-2) s(-1), respectively, so as to phenomenologically fit the time course. A second luminescence peak appears after around 350 s. The dependence of the rate constants on the concentrations of reactants and a viscous reagent revealed that two kinds of reaction contribute the observed time course: (1) an intrinsic reaction by ATP photolyzed from caged-ATP that is already trapped in luciferase; and (2) a diffusion-controlled reaction by free ATP in the buffer solution outside luciferase. Numerical analysis based on reaction kinetics related γ(1) and γ(2) to the rate constants of a three-step reaction model, and accurately described the effects of concentration of reactants and a viscous reagent on the time courses of bioluminescence. Thus, it has been clearly concluded that the binding mode of caged-ATP at the catalytic center of luciferase is very different from that of ATP.

  19. A sensitive aptasensor for colorimetric detection of adenosine triphosphate based on the protective effect of ATP-aptamer complexes on unmodified gold nanoparticles.

    Science.gov (United States)

    Huo, Yuan; Qi, Liang; Lv, Xiao-Jun; Lai, Ting; Zhang, Jing; Zhang, Zhi-Qi

    2016-04-15

    Adenosine triphosphate (ATP) is the most direct source of energy in organisms. This study is the first to demonstrate that ATP-aptamer complexes provide greater protection for unmodified gold nanoparticles (AuNPs) against salt-induced aggregation than either aptamer or ATP alone. This protective effect was confirmed using transmission electron microscopy, dynamic light scattering, Zeta potential measurement, and fluorescence polarization techniques. Utilizing controlled particle aggregation/dispersion as a gauge, a sensitive and selective aptasensor for colorimetric detection of ATP was developed using ATP-binding aptamers as the identification element and unmodified AuNPs as the probe. This aptasensor exhibited a good linear relationship between the absorbance and the logarithm concentration of ATP within a 50-1000 nM range. ATP analogs such as guanosine triphosphate, uridine triphosphate and cytidine triphosphate resulted in little or no interference in the determination of ATP.

  20. Highly Localized Acoustic Streaming and Size-Selective Submicrometer Particle Concentration Using High Frequency Microscale Focused Acoustic Fields.

    Science.gov (United States)

    Collins, David J; Ma, Zhichao; Ai, Ye

    2016-05-17

    Concentration and separation of particles and biological specimens are fundamental functions of micro/nanofluidic systems. Acoustic streaming is an effective and biocompatible way to create rapid microscale fluid motion and induce particle capture, though the >100 MHz frequencies required to directly generate acoustic body forces on the microscale have traditionally been difficult to generate and localize in a way that is amenable to efficient generation of streaming. Moreover, acoustic, hydrodynamic, and electrical forces as typically applied have difficulty manipulating specimens in the submicrometer regime. In this work, we introduce highly focused traveling surface acoustic waves (SAW) at high frequencies between 193 and 636 MHz for efficient and highly localized production of acoustic streaming vortices on microfluidic length scales. Concentration occurs via a novel mechanism, whereby the combined acoustic radiation and streaming field results in size-selective aggregation in fluid streamlines in the vicinity of a high-amplitude acoustic beam, as opposed to previous acoustic radiation induced particle concentration where objects typically migrate toward minimum pressure locations. Though the acoustic streaming is induced by a traveling wave, we are able to manipulate particles an order of magnitude smaller than possible using the traveling wave force alone. We experimentally and theoretically examine the range of particle sizes that can be captured in fluid streamlines using this technique, with rapid particle concentration demonstrated down to 300 nm diameters. We also demonstrate that locations of trapping and concentration are size-dependent, which is attributed to the combined effects of the acoustic streaming and acoustic forces.

  1. Thermodynamics of proton transport coupled ATP synthesis.

    Science.gov (United States)

    Turina, Paola; Petersen, Jan; Gräber, Peter

    2016-06-01

    The thermodynamic H(+)/ATP ratio of the H(+)-ATP synthase from chloroplasts was measured in proteoliposomes after energization of the membrane by an acid base transition (Turina et al. 2003 [13], 418-422). The method is discussed, and all published data obtained with this system are combined and analyzed as a single dataset. This meta-analysis led to the following results. 1) At equilibrium, the transmembrane ΔpH is energetically equivalent to the transmembrane electric potential difference. 2) The standard free energy for ATP synthesis (reference reaction) is ΔG°(ref)=33.8±1.3kJ/mol. 3) The thermodynamic H(+)/ATP ratio, as obtained from the shift of the ATP synthesis equilibrium induced by changing the transmembrane ΔpH (varying either pH(in) or pH(out)) is 4.0±0.1. The structural H(+)/ATP ratio, calculated from the ratio of proton binding sites on the c-subunit-ring in F(0) to the catalytic nucleotide binding sites on the β-subunits in F(1), is c/β=14/3=4.7. We infer that the energy of 0.7 protons per ATP that flow through the enzyme, but do not contribute to shifting the ATP/(ADP·Pi) ratio, is used for additional processes within the enzyme, such as activation, and/or energy dissipation, due e.g. to internal uncoupling. The ratio between the thermodynamic and the structural H(+)/ATP values is 0.85, and we conclude that this value represents the efficiency of the chemiosmotic energy conversion within the chloroplast H(+)-ATP synthase.

  2. Reduction of Temperature in Concentrator Photovoltaic Module Using Coating with High Thermal Emissivity and Conductivity

    Directory of Open Access Journals (Sweden)

    Nakamura Satoshi

    2016-01-01

    Full Text Available The temperature of solar cells considerably increases under light-concentrating operations, and the conversion efficiency of solar cells decreases with increasing temperature. It is very important to reduce the cell temperature in concentrator photovoltaic modules. The thermal radiation layers with high thermal emissivity and thermal conductivity was coated on the aluminum samples which is used for the chassis of concentrator photovoltaic and the effect was evaluated under the conditions with wind. The temperature of sample with coating showed lower temperature than that without coating. In the condition with wind, the coated sample with high thermal emissivity and high thermal conductivity showed the lowest temperature due to the effect of thermal radiation and thermal conduction.

  3. Experimental Study of Sorbitol Production by Zymomonas mobilis in High Sucrose Concentration

    Directory of Open Access Journals (Sweden)

    Rui Sérgio dos Santos Ferreira da Silva

    2004-01-01

    Full Text Available The sorbitol presents several industrial applications and its conventional production is of high cost and low yield. Sorbitol production by Zymomonas mobilis production has attracted attention as both production cost and environmental impact are low. The sorbitol plays an osmo-protective rule so that its production is promoted by high sugar concentrations. This work has evaluated the effect of high sucrose concentration in the sorbitol production. The raise of sucrose concentration from 100 to 300g/ L caused an increase in the sorbitol production from 4,979 to 20,633g/l. Statistic analysis showed that 95,5% in the variation in sorbitol production can be explained.

  4. Interaction between ATP, metal ions, glycine, and several minerals

    Science.gov (United States)

    Rishpon, J.; Ohara, P. J.; Lawless, J. G.; Lahav, N.

    1982-01-01

    Interactions between ATP, glycine and montmorillonite and kaolinite clay minerals in the presence of various metal cations are investigated. The adsorption of adenine nucleotides on clays and Al(OH)3 was measured as a function of pH, and glycine condensation was followed in the presence of ATP, ZnCl2, MgCl2 and either kaolinite or montmorillonite. The amounts of ATP and ADP adsorbed are found to decrease with increasing Ph, and to be considerably enhanced in experiments with Mg(2+)- and Zn(2+)-montmorillonite with respect to Na(+)-montmorillonite. The effects of divalent cations are less marked in kaolinite. Results for Al(OH)3 show the importance of adsorption at clay platelet edges at high pH. The decomposition of ATP during drying at high temperature is observed to be inhibited by small amounts of clay, vacuum, or Mg(2+) or Zn(2+) ions, and to be accompanied by peptide formation in the presence of glycine. Results suggest the importance of Zn(2+) and Mg(2+) in chemical evolution.

  5. Misconceptions on ATP thermodynamic role in cellular processes

    Directory of Open Access Journals (Sweden)

    R. M. Martins

    2011-04-01

    Full Text Available The occurrence and permanence of misconceptions have negative implications on the learning processes  since it impairs theconstruction of significant learning. Misconceptions correction is a complex task due to the difficulties intheir detection and high resistance to their removal. The mainobjective of the present work was to investigate misconceptions about the thermodynamic role of theATP in cellular processes. Tests were realized with high school (HS, undergraduate (UG and graduate students involved in PhD programs (G. In this survey students answered a 15 item questionnaire dealing with the ATP role as the cellular energy source. The stability of such misconceptionswere verified: one result shows that 68% HS, 92% UG and 91% G students state that the energy from ATP hydrolysis is responsiblefor driving cellular processes. Overall results show that students carry misconceptions on basic thermodynamic concepts such as energy transfer and chemical reactions spontaneity. One source of the prevalence of the discussed misconceptions aretextbooks, where schemes, figures and even text early introducefalse concepts on the ATP role.

  6. The ability of Typha domingensis to accumulate and tolerate high concentrations of Cr, Ni, and Zn.

    Science.gov (United States)

    Mufarrege, M M; Hadad, H R; Di Luca, G A; Maine, M A

    2015-01-01

    The tolerance and removal efficiency of Typha domingensis exposed to high concentrations of Cr, Ni, and Zn in single and combined treatments were studied. Sediment and two plants were disposed in each plastic reactor. The treatments were 100 and 500 mg L(-1) of Cr, Ni, and Zn (single solutions); 100 mg L(-1) Cr + Ni + Zn (multi-metal solutions) and 500 mg L(-1) Cr + Ni + Zn (multi-metal solutions); and a control. Even though the concentrations studied were extremely high, simulating an accidental metal dump, the three metals were efficiently removed from water. The highest removal was registered for Cr. The presence of other metals favored Cr and did not favor Ni and Zn removal from water. After 25 days, senescence and chlorosis of plants were observed in Ni and Comb500 treatments, while Cr and Zn only caused growth inhibition. T. domingensis accumulated high metal concentrations in tissues. The roots showed higher metal concentration than submerged parts of leaves. Cr translocation to aerial parts was enhanced by the presence of Ni and Zn. Our results demonstrate that in the case of an accidental dump of high Cr, Ni, and Zn concentrations, a wetland system dominated by T. domingensis is able to retain metals, and the macrophyte is able to tolerate them the time necessary to remove them from water. Thus, the environment will be preserved since the wetland would act as a cushion.

  7. Apoptosis Induced by High Concentrations of Nicotinamide in Tobacco Suspension Cells

    Institute of Scientific and Technical Information of China (English)

    张贵友; 朱瑞宇; 戴尧仁

    2004-01-01

    As an inhibitor of poly(ADP-ribose) polymerase (PARP), nicotinamide has a restraining effect on apoptosis at certain low concentrations. In our present study, apoptosis induced by high concentrations of nicotinamide was observed in tobacco suspension cells. When cells were preincubated with 250 mmol/L nicotinamide for 24 h, the hallmarks of apoptosis were detected, including DNA fragments increasing in size by multiples of 180-200 bp, the condensation and peripheral distribution of nuclear chromatin, and a positive reaction to the TUNEL assay. At the same time, the degradation of PARP and the reduction in the potential of the inner membrane of mitochondria appeared in apoptotic cells induced by high concentrations of nicotinamide. This result indicates that apoptosis induced by high concentrations of nicotinamide is associated with caspase-3-like activity and with the opening of mitochondrial permeability pores. These results partially support the hypothesis that high concentrations of PARP inhibitor could force cells to enter an apoptotic pathway by delay of DNA repair in replicating cells.

  8. Knowledge discovery from high-frequency stream nitrate concentrations: hydrology and biology contributions

    Science.gov (United States)

    Aubert, Alice H.; Thrun, Michael C.; Breuer, Lutz; Ultsch, Alfred

    2016-01-01

    High-frequency, in-situ monitoring provides large environmental datasets. These datasets will likely bring new insights in landscape functioning and process scale understanding. However, tailoring data analysis methods is necessary. Here, we detach our analysis from the usual temporal analysis performed in hydrology to determine if it is possible to infer general rules regarding hydrochemistry from available large datasets. We combined a 2-year in-stream nitrate concentration time series (time resolution of 15 min) with concurrent hydrological, meteorological and soil moisture data. We removed the low-frequency variations through low-pass filtering, which suppressed seasonality. We then analyzed the high-frequency variability component using Pareto Density Estimation, which to our knowledge has not been applied to hydrology. The resulting distribution of nitrate concentrations revealed three normally distributed modes: low, medium and high. Studying the environmental conditions for each mode revealed the main control of nitrate concentration: the saturation state of the riparian zone. We found low nitrate concentrations under conditions of hydrological connectivity and dominant denitrifying biological processes, and we found high nitrate concentrations under hydrological recession conditions and dominant nitrifying biological processes. These results generalize our understanding of hydro-biogeochemical nitrate flux controls and bring useful information to the development of nitrogen process-based models at the landscape scale. PMID:27572284

  9. Experimental studies on resistance characteristics of high concentration red mud in pipeline transport

    Institute of Scientific and Technical Information of China (English)

    WANG Xing; QU Yuan-yuan; HU Wei-wei; CHEN Jie; ZHAO Xue-yi; WU Miao

    2008-01-01

    Red mud will flow in paste form under high pressure during pipeline transport. It belongs to a two-phase flow of materials with high viscosity and a high concentration of non-sedimentation, homogeneous solid-liquids. In pipeline transport, its resistance char-acteristics will be influenced by such factors as grain size, velocity, concentration, density,grain composition and pipe diameter etc.. With the independently developed small-sized tube-type pressure resistance test facility, studied the resistance characteristics of red mud concerning the three influencing factors, paste concentration, velocity and pipe diameter,which attract the most attention in projects. The fine grain size of the red mud is d50=13.02 μm. According to the experimental results, the pressure loss in transport will in-crease along with the increase of velocity and will fall along with the increase of pipe di-ameter. A 1% difference in paste concentration will result in a 50%~100% difference in pipeline resistance loss. These experimental data is hoped to be direct guidance to the design of high concentration and viscous material pipeline transport system.

  10. Knowledge discovery from high-frequency stream nitrate concentrations: hydrology and biology contributions.

    Science.gov (United States)

    Aubert, Alice H; Thrun, Michael C; Breuer, Lutz; Ultsch, Alfred

    2016-08-30

    High-frequency, in-situ monitoring provides large environmental datasets. These datasets will likely bring new insights in landscape functioning and process scale understanding. However, tailoring data analysis methods is necessary. Here, we detach our analysis from the usual temporal analysis performed in hydrology to determine if it is possible to infer general rules regarding hydrochemistry from available large datasets. We combined a 2-year in-stream nitrate concentration time series (time resolution of 15 min) with concurrent hydrological, meteorological and soil moisture data. We removed the low-frequency variations through low-pass filtering, which suppressed seasonality. We then analyzed the high-frequency variability component using Pareto Density Estimation, which to our knowledge has not been applied to hydrology. The resulting distribution of nitrate concentrations revealed three normally distributed modes: low, medium and high. Studying the environmental conditions for each mode revealed the main control of nitrate concentration: the saturation state of the riparian zone. We found low nitrate concentrations under conditions of hydrological connectivity and dominant denitrifying biological processes, and we found high nitrate concentrations under hydrological recession conditions and dominant nitrifying biological processes. These results generalize our understanding of hydro-biogeochemical nitrate flux controls and bring useful information to the development of nitrogen process-based models at the landscape scale.

  11. Small amounts of functional ATP7A protein permit mild phenotype

    DEFF Research Database (Denmark)

    Møller, Lisbeth Birk

    2015-01-01

    Mutations in ATP7A lead to at least three allelic disorders: Menkes disease (MD), Occipital horn syndrome and X-linked distal motor neuropathy. These disorders are mainly seen in male individuals, but a few affected females have been described. More than 400 different mutations have been identified...... concentrations, ATP7A shifts to the post-Golgi compartments or to the plasma membrane to export copper out of the cell. Impaired copper-regulation trafficking has been observed for ATP7A mutants, but its impact on the clinical outcome is not clear. The major problem in patients with MD seems to be insufficient...

  12. Visual and Plasmon Resonance Absorption Sensor for Adenosine Triphosphate Based on the High Affinity between Phosphate and Zr(IV).

    Science.gov (United States)

    Qi, Wenjing; Liu, Zhongyuan; Zhang, Wei; Halawa, Mohamed Ibrahim; Xu, Guobao

    2016-10-12

    Zr(IV) can form phosphate and Zr(IV) (-PO₃(2-)-Zr(4+)-) complex owing to the high affinity between Zr(IV) with phosphate. Zr(IV) can induce the aggregation of gold nanoparticles (AuNPs), while adenosine triphosphate(ATP) can prevent Zr(IV)-induced aggregation of AuNPs. Herein, a visual and plasmon resonance absorption (PRA)sensor for ATP have been developed using AuNPs based on the high affinity between Zr(IV)with ATP. AuNPs get aggregated in the presence of certain concentrations of Zr(IV). After the addition of ATP, ATP reacts with Zr(IV) and prevents AuNPs from aggregation, enabling the detection of ATP. Because of the fast interaction of ATP with Zr(IV), ATP can be detected with a detection limit of 0.5 μM within 2 min by the naked eye. Moreover, ATP can be detected by the PRA technique with higher sensitivity. The A520nm/A650nm values in PRA spectra increase linearly with the concentrations of ATP from 0.1 μM to 15 μM (r = 0.9945) with a detection limit of 28 nM. The proposed visual and PRA sensor exhibit good selectivity against adenosine, adenosine monophosphate, guanosine triphosphate, cytidine triphosphate and uridine triphosphate. The recoveries for the analysis of ATP in synthetic samples range from 95.3% to 102.0%. Therefore, the proposed novel sensor for ATP is promising for real-time or on-site detection of ATP.

  13. Visual and Plasmon Resonance Absorption Sensor for Adenosine Triphosphate Based on the High Affinity between Phosphate and Zr(IV)

    Science.gov (United States)

    Qi, Wenjing; Liu, Zhongyuan; Zhang, Wei; Halawa, Mohamed Ibrahim; Xu, Guobao

    2016-01-01

    Zr(IV) can form phosphate and Zr(IV) (–PO32−–Zr4+–) complex owing to the high affinity between Zr(IV) with phosphate. Zr(IV) can induce the aggregation of gold nanoparticles (AuNPs), while adenosine triphosphate(ATP) can prevent Zr(IV)-induced aggregation of AuNPs. Herein, a visual and plasmon resonance absorption (PRA)sensor for ATP have been developed using AuNPs based on the high affinity between Zr(IV)with ATP. AuNPs get aggregated in the presence of certain concentrations of Zr(IV). After the addition of ATP, ATP reacts with Zr(IV) and prevents AuNPs from aggregation, enabling the detection of ATP. Because of the fast interaction of ATP with Zr(IV), ATP can be detected with a detection limit of 0.5 μM within 2 min by the naked eye. Moreover, ATP can be detected by the PRA technique with higher sensitivity. The A520nm/A650nm values in PRA spectra increase linearly with the concentrations of ATP from 0.1 μM to 15 μM (r = 0.9945) with a detection limit of 28 nM. The proposed visual and PRA sensor exhibit good selectivity against adenosine, adenosine monophosphate, guanosine triphosphate, cytidine triphosphate and uridine triphosphate. The recoveries for the analysis of ATP in synthetic samples range from 95.3% to 102.0%. Therefore, the proposed novel sensor for ATP is promising for real-time or on-site detection of ATP. PMID:27754349

  14. Visual and Plasmon Resonance Absorption Sensor for Adenosine Triphosphate Based on the High Affinity between Phosphate and Zr(IV

    Directory of Open Access Journals (Sweden)

    Wenjing Qi

    2016-10-01

    Full Text Available Zr(IV can form phosphate and Zr(IV (–PO32−–Zr4+– complex owing to the high affinity between Zr(IV with phosphate. Zr(IV can induce the aggregation of gold nanoparticles (AuNPs, while adenosine triphosphate(ATP can prevent Zr(IV-induced aggregation of AuNPs. Herein, a visual and plasmon resonance absorption (PRAsensor for ATP have been developed using AuNPs based on the high affinity between Zr(IVwith ATP. AuNPs get aggregated in the presence of certain concentrations of Zr(IV. After the addition of ATP, ATP reacts with Zr(IV and prevents AuNPs from aggregation, enabling the detection of ATP. Because of the fast interaction of ATP with Zr(IV, ATP can be detected with a detection limit of 0.5 μM within 2 min by the naked eye. Moreover, ATP can be detected by the PRA technique with higher sensitivity. The A520nm/A650nm values in PRA spectra increase linearly with the concentrations of ATP from 0.1 μM to 15 μM (r = 0.9945 with a detection limit of 28 nM. The proposed visual and PRA sensor exhibit good selectivity against adenosine, adenosine monophosphate, guanosine triphosphate, cytidine triphosphate and uridine triphosphate. The recoveries for the analysis of ATP in synthetic samples range from 95.3% to 102.0%. Therefore, the proposed novel sensor for ATP is promising for real-time or on-site detection of ATP.

  15. Extracellular ATP in the Exocrine Pancreas – ATP Release, Signalling and Metabolism

    DEFF Research Database (Denmark)

    Kowal, Justyna Magdalena

    ATP plays an important role as an autocrine/paracrine signalling molecule, being released from a number of tissues, in response to physiological and pathophysiological stimuli. Released ATP induces Ca2+ - and/or cAMP - dependent cellular responses via activation of ubiquitously expressed P2X and P2......Y receptors. Previously, our group has shown that cholinergic stimulation of acini caused ATP release into ducts and ATP is an important regulator of ductal functions by being involved in ion and fluid secretion. Pancreatic duct cells are exposed to a number of stimuli, well known to induce ATP...... release. So far, the contribution of duct cells in purinergic signalling has never been studied. This work presents that both acinar and duct cells are sources of extracellular ATP in the exocrine pancreas. Here we show that duct cells release ATP in response to several physiological...

  16. Extracellular ATP in the Exocrine Pancreas – ATP Release, Signalling and Metabolism

    DEFF Research Database (Denmark)

    Kowal, Justyna Magdalena

    ATP plays an important role as an autocrine/paracrine signalling molecule, being released from a number of tissues, in response to physiological and pathophysiological stimuli. Released ATP induces Ca2+ - and/or cAMP - dependent cellular responses via activation of ubiquitously expressed P2X and P2......Y receptors. Previously, our group has shown that cholinergic stimulation of acini caused ATP release into ducts and ATP is an important regulator of ductal functions by being involved in ion and fluid secretion. Pancreatic duct cells are exposed to a number of stimuli, well known to induce ATP...... release. So far, the contribution of duct cells in purinergic signalling has never been studied. This work presents that both acinar and duct cells are sources of extracellular ATP in the exocrine pancreas. Here we show that duct cells release ATP in response to several physiological...

  17. High surface hole concentration p-type GaN using Mg implantation

    CERN Document Server

    Long Tao; Zhang Guo Yi

    2001-01-01

    Mg ions were implanted on Mg-doped GaN grown by metalorganic chemical vapor deposition (MOCVD). The p-type GaN was achieved with high hole concentration (8.28 x 10 sup 1 sup 7 cm sup - sup 3) conformed by Van derpauw Hall measurement after annealing at 800 degree C for 1 h. this is the first experimental report of Mg implantation on Mg-doped GaN and achieving p-type GaN with high surface hole concentration

  18. ATP synthesis in rod outer segments of bovine retina by the reversal of the disk Ca(2+) pump.

    Science.gov (United States)

    Pepe, I M; Panfoli, I; Notari, L; Morelli, A

    2000-02-16

    Purified disk membranes from rod outer segments of the bovine retina were able to synthesize ATP with a maximal activity (about 52 nmoles ATP/min/mg of protein) at physiological calcium concentrations. This activity was inhibited by vanadate or thapsigargin but not by oligomycin, suggesting the reversal functioning of the disk Ca(2+)-ATPase, which would act as a ATP synthesizer at the expense of the calcium gradient between the disks and the cytoplasm of the rod outer segment. The results are discussed in terms of the need of an immediate source of ATP on the disk membranes where the energy is required to supply the rapid reactions of the photoreception processes.

  19. Adenosine triphosphate (ATP) reduces amyloid-β protein misfolding in vitro.

    Science.gov (United States)

    Coskuner, Orkid; Murray, Ian V J

    2014-01-01

    Alzheimer's disease (AD) is a devastating disease of aging that initiates decades prior to clinical manifestation and represents an impending epidemic. Two early features of AD are metabolic dysfunction and changes in amyloid-β protein (Aβ) levels. Since levels of ATP decrease over the course of the disease and Aβ is an early biomarker of AD, we sought to uncover novel linkages between the two. First and remarkably, a GxxxG motif is common between both Aβ (oligomerization motif) and nucleotide binding proteins (Rossmann fold). Second, ATP was demonstrated to protect against Aβ mediated cytotoxicity. Last, there is structural similarity between ATP and amyloid binding/inhibitory compounds such as ThioT, melatonin, and indoles. Thus, we investigated whether ATP alters misfolding of the pathologically relevant Aβ42. To test this hypothesis, we performed computational and biochemical studies. Our computational studies demonstrate that ATP interacts strongly with Tyr10 and Ser26 of Aβ fibrils in solution. Experimentally, both ATP and ADP reduced Aβ misfolding at physiological intracellular concentrations, with thresholds at ~500 μM and 1 mM respectively. This inhibition of Aβ misfolding is specific; requiring Tyr10 of Aβ and is enhanced by magnesium. Last, cerebrospinal fluid ATP levels are in the nanomolar range and decreased with AD pathology. This initial and novel finding regarding the ATP interaction with Aβ and reduction of Aβ misfolding has potential significance to the AD field. It provides an underlying mechanism for published links between metabolic dysfunction and AD. It also suggests a potential role of ATP in AD pathology, as the occurrence of misfolded extracellular Aβ mirrors lowered extracellular ATP levels. Last, the findings suggest that Aβ conformation change may be a sensor of metabolic dysfunction.

  20. The molecular motor F-ATP synthase is targeted by the tumoricidal protein HAMLET.

    Science.gov (United States)

    Ho, James; Sielaff, Hendrik; Nadeem, Aftab; Svanborg, Catharina; Grüber, Gerhard

    2015-05-22

    HAMLET (human alpha-lactalbumin made lethal to tumor cells) interacts with multiple tumor cell compartments, affecting cell morphology, metabolism, proteasome function, chromatin structure and viability. This study investigated if these diverse effects of HAMLET might be caused, in part, by a direct effect on the ATP synthase and a resulting reduction in cellular ATP levels. A dose-dependent reduction in cellular ATP levels was detected in A549 lung carcinoma cells, and by confocal microscopy, co-localization of HAMLET with the nucleotide-binding subunits α (non-catalytic) and β (catalytic) of the energy converting F1F0 ATP synthase was detected. As shown by fluorescence correlation spectroscopy, HAMLET binds to the F1 domain of the F1F0 ATP synthase with a dissociation constant (KD) of 20.5μM. Increasing concentrations of the tumoricidal protein HAMLET added to the enzymatically active α3β3γ complex of the F-ATP synthase lowered its ATPase activity, demonstrating that HAMLET binding to the F-ATP synthase effects the catalysis of this molecular motor. Single-molecule analysis was applied to study HAMLET-α3β3γ complex interaction. Whereas the α3β3γ complex of the F-ATP synthase rotated in a counterclockwise direction with a mean rotational rate of 3.8±0.7s(-1), no rotation could be observed in the presence of bound HAMLET. Our findings suggest that direct effects of HAMLET on the F-ATP synthase may inhibit ATP-dependent cellular processes. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Ionotropic P2X ATP Receptor Channels Mediate Purinergic Signaling in Mouse Odontoblasts

    Science.gov (United States)

    Shiozaki, Yuta; Sato, Masaki; Kimura, Maki; Sato, Toru; Tazaki, Masakazu; Shibukawa, Yoshiyuki

    2017-01-01

    ATP modulates various functions in the dental pulp cells, such as intercellular communication and neurotransmission between odontoblasts and neurons, proliferation of dental pulp cells, and odontoblast differentiation. However, functional expression patterns and their biophysical properties of ionotropic ATP (P2X) receptors (P2X1–P2X7) in odontoblasts were still unclear. We examined these properties of P2X receptors in mouse odontoblasts by patch-clamp recordings. K+-ATP, nonselective P2X receptor agonist, induced inward currents in odontoblasts in a concentration-dependent manner. K+-ATP-induced currents were inhibited by P2X4 and P2X7 selective inhibitors (5-BDBD and KN62, respectively), while P2X1 and P2X3 inhibitors had no effects. P2X7 selective agonist (BzATP) induced inward currents dose-dependently. We could not observe P2X1, 2/3, 3 selective agonist (αβ-MeATP) induced currents. Amplitudes of K+-ATP-induced current were increased in solution without extracellular Ca2+, but decreased in Na+-free extracellular solution. In the absence of both of extracellular Na+ and Ca2+, K+-ATP-induced currents were completely abolished. K+-ATP-induced Na+ currents were inhibited by P2X7 inhibitor, while the Ca2+ currents were sensitive to P2X4 inhibitor. These results indicated that odontoblasts functionally expressed P2X4 and P2X7 receptors, which might play an important role in detecting extracellular ATP following local dental pulp injury. PMID:28163685

  2. High-concentration zeta potential measurements using light-scattering techniques.

    Science.gov (United States)

    Kaszuba, Michael; Corbett, Jason; Watson, Fraser Mcneil; Jones, Andrew

    2010-09-28

    Zeta potential is the key parameter that controls electrostatic interactions in particle dispersions. Laser Doppler electrophoresis is an accepted method for the measurement of particle electrophoretic mobility and hence zeta potential of dispersions of colloidal size materials. Traditionally, samples measured by this technique have to be optically transparent. Therefore, depending upon the size and optical properties of the particles, many samples will be too concentrated and will require dilution. The ability to measure samples at or close to their neat concentration would be desirable as it would minimize any changes in the zeta potential of the sample owing to dilution. However, the ability to measure turbid samples using light-scattering techniques presents a number of challenges. This paper discusses electrophoretic mobility measurements made on turbid samples at high concentration using a novel cell with reduced path length. Results are presented on two different sample types, titanium dioxide and a polyurethane dispersion, as a function of sample concentration. For both of the sample types studied, the electrophoretic mobility results show a gradual decrease as the sample concentration increases and the possible reasons for these observations are discussed. Further, a comparison of the data against theoretical models is presented and discussed. Conclusions and recommendations are made from the zeta potential values obtained at high concentrations.

  3. High Iridium concentration of alkaline rocks of Deccan and implications to K/T boundary

    Indian Academy of Sciences (India)

    P N Shukla; N Bhandari; Anirban Das; A D Shukla; J S Ray

    2001-06-01

    We report here an unusually high concentration of iridium in some alkali basalts and alkaline rocks of Deccan region having an age of about 65Ma, similar to the age of the Cretaceous-Tertiary boundary. The alkali basalts of Anjar, in the western periphery of Deccan province, have irid-ium concentration as high as 178pg/g whereas the alkaline rocks and basalts associated with the Amba Dongar carbonatite complex have concentrations ranging between 8 and 80 pg/g. Some of these values are more than an order of magnitude higher than the concentration in the tholeiiticbasalts of Deccan, indicating the significance of alkaline magmatism in the iridium inventory at the Cretaceous-Tertiary boundary. Despite higher concentration, their contribution to the global inventory of iridium in the Cretaceous-Tertiary boundary clays remains small. The concentration of iridium in uorites from Amba Dongar was found to be < 30 pg/g indicating that iridium is not incorporated during their formation in hydrothermal activity.

  4. Potential risk of mitomycin Cat high concentrations on peripheral nerve structure

    Institute of Scientific and Technical Information of China (English)

    Tao Sui; Jinhong Zhang; Shihao Du; Changhui Su; Jun Que; Xiaojian Cao

    2014-01-01

    Although the local application of mitomycin C may prevent epidural adhesion after laminectomy, mitomycin C can induce neurotoxicity in optic and acoustic nerves at high concentrations. To determine the safe concentration range for mitomycin C, cotton pads soaked with mitomycin C at different concentrations (0.1, 0.3, 0.5, and 0.7 mg/mL) were immediately applied for 5 minutes to the operation area of rats that had undergone laminectomy at L1. Rat sciatic nerves, instead of dorsal nerves, were used in this study. The results showed that mitomycin C at 0.1-0.5 mg/mL did not damage the structure and function of the sciatic nerve, while at 0.7 mg/mL, mitomycin C signiifcantly reduced the thickness of the sciatic nerve myelin sheath compared with lower concen-trations, though no functional change was found. These experimental ifndings indicate that the local application of mitomycin C at low concentrations is safe to prevent scar adhesion following laminectomy, but that mitomycin C at high concentrations (>0.7 mg/mL) has potential safety risks to peripheral nerve structures.

  5. Vitamin E Concentrations in Adults with HIV/AIDS on Highly Active Antiretroviral Therapy

    Directory of Open Access Journals (Sweden)

    Daniella J. Itinoseki Kaio

    2014-09-01

    Full Text Available HIV/AIDS patients are probably more predisposed to vitamin E deficiency, considering that they are more exposed to oxidative stress. Additionally, there are an extensive number of drugs in the highly active antiretroviral therapy (HAART regimens that may interfere with vitamin E concentrations. The objective of this study was to compare serum concentrations of alpha-tocopherol in 182 HIV/AIDS patients receiving different HAART regimens. The patients were divided into three groups according to regimen: nucleoside analog reverse-transcriptase inhibitors (NRTIs + non-nucleoside analog reverse-transcriptase inhibitors (NNRTIs; NRTIs + protease inhibitors + ritonavir; NRTIs + other classes. Alpha-tocopherol was assessed by high-performance liquid chromatography. Multiple linear regression analysis was used to evaluate the effects of HAART regimen, time of use, and compliance with the regimen on alpha-tocopherol concentrations. Alpha-tocopherol concentrations were on average 4.12 μmol/L lower for the NRTIs + other classes regimen when compared to the NRTIs + NNRTIs regimen (p = 0.037. A positive association (p < 0.001 was observed between alpha-tocopherol and cholesterol concentrations, a finding due, in part, to the relationship between liposoluble vitamins and lipid profile. This study demonstrated differences in alpha-tocopherol concentrations between patients using different HAART regimens, especially regimens involving the use of new drugs. Long-term prospective cohort studies are needed to monitor vitamin E status in HIV/AIDS patients since the beginning of treatment.

  6. Characterization of high-milk-protein powders upon rehydration under various salt concentrations.

    Science.gov (United States)

    Hussain, R; Gaiani, C; Aberkane, L; Scher, J

    2011-01-01

    Rehydration of native micellar casein and native whey isolate protein powders was followed in different ionic environments. Solutions of NaCl and CaCl2 in the concentration range of 0 to 12% (wt%) were used as rehydration media. The rehydration profiles obtained were interpreted in terms of wetting, swelling, and dispersion stages by using a turbidity method. Two behaviors were observed depending on the salt concentration. For native micellar casein powder, a significant change was observed between 3 and 6% NaCl and between 0.75 and 1.5% CaCl2. The first behavior (low salt concentration) presents a typical rehydration profile: quick wetting, swelling, and long dispersion stage. The dispersion stage of the second behavior (high salt concentration) was significantly shortened, indicating a strong modification of the protein backbone. The rehydration of whey protein powder was less influenced by salts. At low salt concentrations, a typical profile for whey powders was observed: wetting with lump formation and no swelling followed by a quick dispersion. At high CaCl2 concentrations, no turbidity stabilization was observed, indicating a possible protein unfolding and denaturation. Additionally, the changes in secondary structures of the 2 proteins upon salt increase were followed by Fourier transform infrared spectroscopy and confirmed the different profiles observed. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  7. High hydrostatic pressure enables almost 100% refolding of recombinant human ciliary neurotrophic factor from inclusion bodies at high concentration.

    Science.gov (United States)

    Wang, Qi; Liu, Yongdong; Zhang, Chun; Guo, Fangxia; Feng, Cui; Li, Xiunan; Shi, Hong; Su, Zhiguo

    2017-05-01

    Protein refolding from inclusion bodies (IBs) often encounters a problem of low recovery at high protein concentration. In this study, we demonstrated that high hydrostatic pressure (HHP) could simultaneously achieve high refolding concentration and high refolding yield for IBs of recombinant human ciliary neurotrophic factor (rhCNTF), a potential therapeutic for neurodegenerative diseases. The use of dilution refolding obtained 18% recovery at 3 mg/mL, even in the presence of 4 M urea. In contrast, HHP refolding could efficiently increase the recovery up to almost 100% even at 4 mg/mL. It was found that in the dilution, hydrophobic aggregates were the off-path products and their amount increased with the protein concentration. However, HHP could effectively minimize the formation of hydrophobic aggregates, leading to almost complete conversion of the rhCNTF IBs to the correct configuration. The stable operation range of concentration is 0.5-4.0 mg/mL, in which the refolding yield was almost 100%. Compared with the literatures where HHP failed to increase the refolding yield beyond 90%, the reason could be attributed to the structural difference that rhCNTF has no disulfide bond and is a monomeric protein. After purification by one-step of anionic chromatography, the purity of rhCNTF reached 95% with total process recovery of 54.1%. The purified rhCNTF showed similar structure and in vitro bioactivity to the native species. The whole process featured integration of solubilization/refolding, a high refolding yield of 100%, a high concentration of 4 mg/mL, and a simple chromatography to ensure a high productivity. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. High Frequency Measurements of Methane Concentrations and Carbon Isotopes at a Marsh and Landfill

    Science.gov (United States)

    Mortazavi, B.; Wilson, B.; Chanton, J.; Eller, K.; Dong, F.; Baer, D. S.; Gupta, M.; Dzwonkowski, B.

    2012-12-01

    High frequency measurements of methane concentrations and carbon isotopes can help constrain the source strengths of methane emitted to the atmosphere. We report here methane concentrations and 13C values measured at 0.5 Hz with cavity enhanced laser absorption spectrometers (Los Gatos Research) deployed at a saltmarsh in Alabama and a landfill in Florida. Methane concentrations and 13C at the saltmarsh were monitored over a 2.5 day time period at 2 m, 0.5 m above the ground as well as from the outflow of a flow-through (2 L) chamber placed on the Spartina alterniflora dominated marsh. A typical measurement cycle included regular samples from two tanks of known methane concentrations and isotopic values and from ambient air samples. Over the 2.5-day measurement period methane concentrations and isotopic ratios at 2 m averaged 1.85 ppm and -43.57‰ (±0.34, 1 SE), respectively. The concentration and isotopic values from the chamber outflow varied from 1.92 to 5.81 ppm and -38.5 to -59.3‰, respectively. Methane flux from the marsh ranged from undetectable to 3.6 mgC m-2hr-1, with high fluxes measured during low tide. The 13δCH4 of the emitted CH4 from the marsh, determined from a mass balance equation using the chamber inflow and outflow concentration and isotopic values ranged from -62.1 to -93.9‰ and averaged -77‰ (±1.25, 1SE). At the landfill ambient methane concentrations and 13C ratios measured over multiple days varied from 4.25 to 11.91 ppm and from -58.81 to -45.12‰, respectively. At higher methane concentrations the δ13C of CH4 was more depleted consistent with previously observed relationship at this site made by more traditional techniques. Over a 30-minute measurement period CH4 concentrations at the landfill could vary by as much as 15 ppm. The high frequency continuous optical measurements with field-deployed instruments provide us with an unprecedented temporal resolution of CH4 concentrations and isotopic ratios. These measurements will

  9. The rise of the photosynthetic rate when light intensity increases is delayed in ndh gene-defective tobacco at high but not at low CO2 concentrations

    Directory of Open Access Journals (Sweden)

    Mercedes eMartin

    2015-02-01

    Full Text Available The 11 plastid ndh genes have hovered frequently on the edge of dispensability, being absent in the plastid DNA of many algae and certain higher plants. We have compared the photosynthetic activity of tobacco (Nicotiana tabacum, cv. Petit Havana with five transgenic lines (ndhF, pr-ndhF, T181D, T181A and ndhF FC and found that photosynthetic performance is impaired in transgenic ndhF-defective tobacco plants at rapidly fluctuating light intensities and higher than ambient CO2 concentrations. In contrast to wild type and ndhF FC, which reach the maximum photosynthetic rate in less than one min when light intensity suddenly increases, ndh defective plants (ndhF and T181A show up to a 5 min delay in reaching the maximum photosynthetic rate at CO2 concentrations higher than the ambient 360 ppm. Net photosynthesis was determined at different CO2 concentrations when sequences of 130, 870, 61, 870 and 130 μmol m−2 s−1 PAR sudden light changes were applied to leaves and photosynthetic efficiency and entropy production were determined as indicators of photosynthesis performance. The two ndh-defective plants, ndhF and T181A, had lower photosynthetic efficiency and higher entropy production than wt, ndhF FC and T181D tobacco plants, containing full functional ndh genes, at CO2 concentrations above 400 ppm. We propose that the Ndh complex improves cyclic electron transport by adjusting the redox level of transporters during the low light intensity stage. In ndhF-defective strains, the supply of electrons through the Ndh complex fails, transporters remain over-oxidized (specially at high CO2 concentrations and the rate of cyclic electron transport is low, impairing the ATP level required to rapidly reach high CO2 fixation rates in the following high light phase. Hence, ndh genes could be dispensable at low but not at high atmospheric concentrations of CO2.

  10. The rise of the photosynthetic rate when light intensity increases is delayed in ndh gene-defective tobacco at high but not at low CO2 concentrations.

    Science.gov (United States)

    Martín, Mercedes; Noarbe, Dolores M; Serrot, Patricia H; Sabater, Bartolomé

    2015-01-01

    The 11 plastid ndh genes have hovered frequently on the edge of dispensability, being absent in the plastid DNA of many algae and certain higher plants. We have compared the photosynthetic activity of tobacco (Nicotiana tabacum, cv. Petit Havana) with five transgenic lines (ΔndhF, pr-ΔndhF, T181D, T181A, and ndhF FC) and found that photosynthetic performance is impaired in transgenic ndhF-defective tobacco plants at rapidly fluctuating light intensities and higher than ambient CO2 concentrations. In contrast to wild type and ndhF FC, which reach the maximum photosynthetic rate in less than 1 min when light intensity suddenly increases, ndh defective plants (ΔndhF and T181A) show up to a 5 min delay in reaching the maximum photosynthetic rate at CO2 concentrations higher than the ambient 360 ppm. Net photosynthesis was determined at different CO2 concentrations when sequences of 130, 870, 61, 870, and 130 μmol m(-2) s(-1) PAR sudden light changes were applied to leaves and photosynthetic efficiency and entropy production (Sg) were determined as indicators of photosynthesis performance. The two ndh-defective plants, ΔndhF and T181A, had lower photosynthetic efficiency and higher Sg than wt, ndhF FC and T181D tobacco plants, containing full functional ndh genes, at CO2 concentrations above 400 ppm. We propose that the Ndh complex improves cyclic electron transport by adjusting the redox level of transporters during the low light intensity stage. In ndhF-defective strains, the supply of electrons through the Ndh complex fails, transporters remain over-oxidized (specially at high CO2 concentrations) and the rate of cyclic electron transport is low, impairing the ATP level required to rapidly reach high CO2 fixation rates in the following high light phase. Hence, ndh genes could be dispensable at low but not at high atmospheric concentrations of CO2.

  11. Lysozyme refolding at high concentration by dilution and size-exclusion chromatography

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    This study of renaturation by dilution and size exclusion chromatography (SEC) addition of urea to improve yield as well as the initial and final protein concentrations showed that although urea decreased the rate of lysozyme refolding, it could suppress protein aggregation to sustain the pathway of correct refolding at high protein concentration; and that there existed an optimum urea concentration in renaturation buffer. Under the above conditions, lysozyme was successfully refolded from initial concentration of up to 40 mg/mL by dilution and 100 mg/mL by SEC, with the yield of the former being more than 40% and that of the latter being 34.8%. Especially, under the condition of 30 min interval time, i.e. τ>2(tR2-tR1), the efficiency was increased by 25% and the renaturation buffer could be recycled for SEC refolding in continuous operation of downstream process.

  12. Determination of adenosine disodium triphosphate (ATP) using norfloxacin-Tb{sup 3+} as a fluorescence probe by spectrofluorimetry

    Energy Technology Data Exchange (ETDEWEB)

    Miao Yanhong [Department of Chemistry, Shandong Normal University, Jinan 250014 (China); College of Science and Technology, Shandong Agriculture University, Taian 271000 (China); Liu Jinkai [Department of Chemistry, Shandong Normal University, Jinan 250014 (China); Hou Faju [Department of Chemistry, Shandong Normal University, Jinan 250014 (China); Jiang Chongqiu [Department of Chemistry, Shandong Normal University, Jinan 250014 (China)]. E-mail: jiangchongqiu@sdnu.edu.cn

    2006-01-15

    A new spectrofluorimetric method was developed for determination of trace amount of adenosine disodium triphosphate (ATP). Using norfloxacin (NFLX)-terbium (Tb{sup 3+}) as a fluorescent probe, in the buffer solution of pH=7.40, ATP can remarkably enhance the fluorescence intensity of the NFLX-Tb{sup 3+} complex at {lambda}=545nm and the enhanced fluorescence intensity of Tb{sup 3+} ion is in proportion to the concentration of ATP. Optimum conditions for the determination of ATP were also investigated. The dynamic range for the determination of ATP is 1.00x10{sup -6}-1.60x10{sup -5}mol/L with detection limit of 4.13x10{sup -8}mol/L. This method is simple, practical and relatively free interference from coexisting substances and can be successfully applied to determination of ATP in samples.

  13. Extracellular ATP induces spikes in cytosolic free Ca(2+) but not in NADPH oxidase activity in neutrophils

    DEFF Research Database (Denmark)

    Brasen, Jens Christian; Olsen, Lars Folke; Hallett, Maurice B

    2011-01-01

    In order to establish whether non-mitochondrial oxidase activity in human neutrophils is tightly related to cytosolic Ca(2+) concentration, we simultaneously measured Ca(2+) oscillations induced by ATP and oxidant production in single adherent neutrophils using confocal microscopy. ATP induced fast...

  14. Extracellular ATP induces spikes in cytosolic free Ca2+ but not in NADH oxidase activity in neutrophils

    DEFF Research Database (Denmark)

    Brasen, Jens Christian; Olsen, Lars Folke; Hallett, Maurice B.

    2011-01-01

    In order to establish whether non-mitochondrial oxidase activity in human neutrophils is tightly related to cytosolic Ca2+ concentration, we simultaneously measured Ca2+ oscillations induced by ATP and oxidant production in single adherent neutrophils using confocal microscopy. ATP induced fast...

  15. Potential of membrane distillation for production of high quality fruit juice concentrate.

    Science.gov (United States)

    Onsekizoglu Bagci, Pelin

    2015-01-01

    Fruit juices are generally concentrated in order to improve the stability during storage and to reduce handling, packaging, and transportation costs. Thermal evaporation is the most widely used technique in industrial fruit juice concentrate production. In addition to high energy consumption, a large part of the characteristics determining the quality of the fresh juice including aroma, color, vitamins, and antioxidants undergoes remarkable alterations through the use of high operation temperatures. Increasing consumer demand for minimally or naturally processed stable products able to retain as much possible the uniqueness of the fresh fruit has engendered a growing interest for development of nonthermal approaches for fruit juice concentration. Among them, membrane distillation (MD) and its variants have attracted much attention for allowing very high concentrations to be reached under atmospheric pressure and temperatures near ambient temperature. This review will provide an overview of the current status and recent developments in the use of MD for concentration of fruit juices. In addition to the most basic concepts of MD variants, crucial suggestions for membrane selection and operating parameters will be presented. Challenges and future trends for industrial adaptation taking into account the possibility of integrating MD with other existing processes will be discussed.

  16. Investigating DOC export dynamics using high-frequency instream concentration measurements

    Science.gov (United States)

    Oosterwoud, Marieke; Keller, Toralf; Musolff, Andreas; Frei, Sven; Park, Ji-Hyung; Fleckenstein, Jan H.

    2014-05-01

    Being able to monitor DOC concentrations using in-situ high frequency measurements makes it possible to better understand concentration-discharge behavior under different hydrological conditions. We developed a UV-Vis probe setup for modified/adapted use under field conditions. The quasi mobile probe setup allows a more flexible probe deployment. New or existing monitoring sites can easily be equipped for quasi-continuous monitoring or measurements can be performed at changing locations, without the need for additional infrastructure. We were able to gather high frequency data on DOC dynamics for one year in two streams in the Harz mountains in Germany. It proved that obtaining accurate DOC concentrations from the UV-Vis probes required frequent maintenance and probe calibration. The advantage of the setup over standard monitoring protocols becomes evident when comparing net exports over a year. In addition to mass improved balance calculations the high-frequency measurements can reveal intricate hysteretic relationships between discharge and concentrations that can provide valuable insights into the hydrologic dynamics and mechanisms that govern the delivery of DOC to the receiving waters. Measurements with similar probes from two additional catchments in Southern Germany and South Korea will be used to illustrate different discharge-concentration relationships and what can be learned from them about the hydrologic mechanisms that control the dynamics of DOC export.

  17. Performance of solmacs, a high PV solar concentrator with efficient optics

    Science.gov (United States)

    Thibert, T.; Hellin, M.-L.; Loicq, J.; Mazy, E.; Jacques, L.; Verstraeten, D.; Gillis, J.-M.; Languy, F.; Emmerechts, C.; Beeckman, E.; Habraken, S.; Lecat, J.-H.

    2012-10-01

    A new solar panel with high concentration photovoltaic technology (x700) has been designed, prototyped and tested in the SOLMACS project. The quality of concentrating optics is a key factor for high module efficiency. Therefore new dedicated PMMA Fresnel lenses were studied and produced by injection molding. Lens design, material and production process were optimized to achieve a high optical yield of 86%. Thorough lens performance assessment in optical laboratory was completed with lifetime UV aging tests. Another important aspect is the thermal control of the hot spot created under the solar cell that receives the concentrated flux of 700 Suns. A dedicated heat spreader was developed to achieve passive thermal control with minimum mass and cost. This was supported by thermal models and tests at both cell and module level. 35% triple junction cells were implemented in the module. Micro-assembly technologies were used for the cell packaging and electrical connections. In support to the research, a continuous solar simulator was designed and built to assess the system performance, both at component and module level. The concentrator developments were integrated in a prototype and tested both indoor with the simulator and outdoor on the CSL solar test platform. The overall efficiency of the PV concentrator module is 28.5%.

  18. Tension Recovery following Ramp-Shaped Release in High-Ca and Low-Ca Rigor Muscle Fibers: Evidence for the Dynamic State of AMADP Myosin Heads in the Absence of ATP

    Science.gov (United States)

    Sugi, Haruo; Yamaguchi, Maki; Ohno, Tetsuo; Kobayashi, Takakazu; Chaen, Shigeru; Okuyama, Hiroshi

    2016-01-01

    During muscle contraction, myosin heads (M) bound to actin (A) perform power stroke associated with reaction, AMADPPi → AM + ADP + Pi. In this scheme, A • M is believed to be a high-affinity complex after removal of ATP. Biochemical studies on extracted protein samples show that, in the AM complex, actin-binding sites are located at both sides of junctional peptide between 50K and 20K segments of myosin heavy chain. Recently, we found that a monoclonal antibody (IgG) to the junctional peptide had no effect on both in vitro actin-myosin sliding and skinned muscle fiber contraction, though it covers the actin-binding sites on myosin. It follows from this that, during muscle contraction, myosin heads do not pass through the static rigor AM configuration, determined biochemically and electron microscopically using extracted protein samples. To study the nature of AM and AMADP myosin heads, actually existing in muscle, we examined mechanical responses to ramp-shaped releases (0.5% of Lo, complete in 5ms) in single skinned rabbit psoas muscle fibers in high-Ca (pCa, 4) and low-Ca (pCa, >9) rigor states. The fibers exhibited initial elastic tension drop and subsequent small but definite tension recovery to a steady level. The tension recovery was present over many minutes in high-Ca rigor fibers, while it tended to decrease quickly in low-Ca rigor fibers. EDTA (10mM, with MgCl2 removed) had no appreciable effect on the tension recovery in high-Ca rigor fibers, while it completely eliminated the tension recovery in low-Ca rigor fibers. These results suggest that the AMADP myosin heads in rigor muscle have long lifetimes and dynamic properties, which show up as the tension recovery following applied release. Possible AM linkage structure in muscle is discussed in connection with the X-ray diffraction pattern from contracting muscle, which is intermediate between resting and rigor muscles. PMID:27583360

  19. Tension Recovery following Ramp-Shaped Release in High-Ca and Low-Ca Rigor Muscle Fibers: Evidence for the Dynamic State of AMADP Myosin Heads in the Absence of ATP.

    Science.gov (United States)

    Sugi, Haruo; Yamaguchi, Maki; Ohno, Tetsuo; Kobayashi, Takakazu; Chaen, Shigeru; Okuyama, Hiroshi

    2016-01-01

    During muscle contraction, myosin heads (M) bound to actin (A) perform power stroke associated with reaction, AMADPPi → AM + ADP + Pi. In this scheme, A • M is believed to be a high-affinity complex after removal of ATP. Biochemical studies on extracted protein samples show that, in the AM complex, actin-binding sites are located at both sides of junctional peptide between 50K and 20K segments of myosin heavy chain. Recently, we found that a monoclonal antibody (IgG) to the junctional peptide had no effect on both in vitro actin-myosin sliding and skinned muscle fiber contraction, though it covers the actin-binding sites on myosin. It follows from this that, during muscle contraction, myosin heads do not pass through the static rigor AM configuration, determined biochemically and electron microscopically using extracted protein samples. To study the nature of AM and AMADP myosin heads, actually existing in muscle, we examined mechanical responses to ramp-shaped releases (0.5% of Lo, complete in 5ms) in single skinned rabbit psoas muscle fibers in high-Ca (pCa, 4) and low-Ca (pCa, >9) rigor states. The fibers exhibited initial elastic tension drop and subsequent small but definite tension recovery to a steady level. The tension recovery was present over many minutes in high-Ca rigor fibers, while it tended to decrease quickly in low-Ca rigor fibers. EDTA (10mM, with MgCl2 removed) had no appreciable effect on the tension recovery in high-Ca rigor fibers, while it completely eliminated the tension recovery in low-Ca rigor fibers. These results suggest that the AMADP myosin heads in rigor muscle have long lifetimes and dynamic properties, which show up as the tension recovery following applied release. Possible AM linkage structure in muscle is discussed in connection with the X-ray diffraction pattern from contracting muscle, which is intermediate between resting and rigor muscles.

  20. Inhibition effects of high calcium concentration on anaerobic biological treatment of MSW leachate.

    Science.gov (United States)

    Xia, Yi; He, Pin-Jing; Pu, Hong-Xia; Lü, Fan; Shao, Li-Ming; Zhang, Hua

    2016-04-01

    With the increasing use of municipal solid waste incineration (MSWI) and more stringent limits on landfilling of organic waste, more MSWI bottom ash is being landfilled, and the proportion of inorganic wastes in landfills is increasing, causing the increased Ca concentrations in landfill leachate. In this research, the inhibition effect of Ca concentration on the anaerobic treatment of landfill leachate was studied using a biochemical methane potential experiment. Slight inhibition of methane production occurred when the addition of Ca concentration was less than 2000 mg/L. When the addition of Ca concentration was between 6000 and 8000 mg/L, methane production was significantly reduced (to 29.4-34.8 % of that produced by the BLK reactor), and the lag phase was increased from 8.55 to 16.32 d. Moreover, when the dosage of Ca concentration increased from zero to 8000 mg/L, reductions in solution Ca concentration increased from 929 to 2611 mg/L, and the proportion of Ca in the residual sludge increased from 22.58 to 46.87 %. Based on the results, when the dosage of Ca concentration was less than 4000 mg/L, the formation of Ca precipitates on the surface of sludge appeared to prevent mass transfer and was the dominant reason for the reduction in methane production and sludge biomass. At higher Ca concentrations (6000-8000 mg/L), the severe inhibition of methane production appeared to be caused by the toxic effect of highly concentrated Ca on sludge as well as mass transfer blockage.

  1. Enhanced Enzymatic Production of Cephalexin at High Substrate Concentration with in situ Product Removal by Complexation

    Directory of Open Access Journals (Sweden)

    Dengchao Li

    2008-01-01

    Full Text Available Cephalexin (CEX was synthesized with 7-amino-3-deacetoxycephalosporanic acid (7-ADCA and D(–-phenylglycine methyl ester (PGME using immobilized penicillin G acylase from Escherichia coli. It was found that substrate concentration and in situ product could remarkably influence the ratio of synthesis to hydrolysis (S/H and the efficiency of CEX synthesis. The optimal ratio of enzyme to substrate was 65 IU/mM 7-ADCA. High substrate concentration improved the 7-ADCA conversion from 61 to 81 % in the process without in situ product removal (ISPR, while in the synthetic process with ISPR, high substrate concentration increased the 7-ADCA conversion from 88 to 98 %. CEX was easily separated from CEX/β-naphthol complex and its purity and overall yield were 99 and 70 %, respectively.

  2. Self-assembling semicrystalline polymer into highly ordered, microscopic concentric rings by evaporation.

    Science.gov (United States)

    Byun, Myunghwan; Hong, Suck Won; Zhu, Lei; Lin, Zhiqun

    2008-04-01

    A drop of semicrystalline polymer, poly(ethylene oxide) (PEO), solution was placed in a restricted geometry consisting of a sphere on a flat substrate (i.e., sphere-on-flat geometry). Upon solvent evaporation from the sphere-on-flat geometry, microscopic concentric rings of PEO with appropriate high molecular weight were produced via controlled, repetitive pinning ("stick") and depinning ("slip") cycles of the contact line. The evaporation-induced concentric rings of PEO exhibited a fibrillar-like surface morphology. Subsequent isothermal crystallization of rings at 40 and 58 degrees C led to the formation of multilayer of flat-on lamellae (i.e., spiral morphology). In between adjacent spirals, depletion zones were developed during crystallization, as revealed by AFM measurements. The present highly ordered, concentric PEO rings may serve as a platform to study cell adhesion and motility, neuron guidance, cell mechanotransduction, and other biological processes.

  3. Chlorinated pesticides (2,4-D and DDT) biodegradation at high concentrations using immobilized Pseudomonas fluorescens.

    Science.gov (United States)

    Santacruz, Germán; Bandala, Erick R; Torres, Luis G

    2005-01-01

    Degradation of two chlorinated pesticides (2,4-D and DDT) using a 54-mL glass column packed with tezontle (a low-cost basaltic scoria) was tested. Bacteria were cultured in YPG (yeast, peptone, and glucose) liquid medium at 32 degrees C. The rich medium was pumped during 24 h through the column to inoculate it. Later, the wasted medium was discharged and the pesticide added. Optical densities, TOC, and pesticide concentration were determined. Pesticide removals for 2,4-D (with initial concentration between 100 and 500 mg/L) were about 99%. DDT removal (at initial concentration of up to 150 mg/L) was as high as 55-99%. TOC removals for 2,4-D was in the 36-87% interval, whereas for DDT they were as high as 36-78%.

  4. A method for analyzing on-line video images of crystallization at high-solid concentrations

    Institute of Scientific and Technical Information of China (English)

    Jian Wan; Cai Y.Ma; Xue Z.Wang

    2008-01-01

    Recent research has demonstrated that on-line video imaging is a very promising technique for monitoring crystallization processes. The bottleneck in applying the technique for real-time closed-loop control is considered as image analysis that needs to be robust, fast and able to handle varied image qualities due to temporal variations of operating conditions such as mixing and solid concentrations. Image analysis at high-solid concentrations turns out to be extremely challenging because crystals tend to overlap or attach to each other and the boundaries between the crystals are usually ambiguous. This paper presents an image segmentation algorithm that can effectively deal with images taken at high-solid concentrations. The method segments crystals attached to each other along the mostly related concave points on the contours of crystal blocks. The detailed procedure is introduced with application to crystallization of L-glutamic acid in a hot-stage reactor.

  5. Highly concentrated phenolic wastewater treatment by heterogeneous and homogeneous photocatalysis: mechanism study by FTIR-ATR.

    Science.gov (United States)

    Araña, J; Tello-Rendón, E; Doña-Rodríguez, J M; Campo, C V; Herrera-Melidán, J A; González-Díaz; Pérez-Peña, J

    2001-01-01

    The degradation of high phenol concentrations (1 g/L) in water solutions by TiO2 photocatalysis and the photo-Fenton reaction has been studied. From the obtained data it may be suggested that degradation of phenol by TiO2-UV takes place onto the catalyst surface by means of peroxo-compounds formation. At low phenol concentrations other mechanism, the insertion of OH. radicals, may be favored. On the other hand, highly concentrated phenol aqueous solutions treatment by the photo-Fenton reaction gives rise to the formation of polyphenolic polymers. These seem to reduce the process rate. Degradation intermediates have been identified by HPLC and FTIR. The FTIR study of the catalyst surface has shown infrared bands attributable to different chemisorbed peroxo-compounds, formates, ortho-formates and carboxylates that can inactivate the catalyst.

  6. High School and Employment Experiences of Vocational and Nonvocational Concentrators of the Idaho High School Graduating Class of 1983.

    Science.gov (United States)

    Riesenberg, Lou E.; Stenberg, Laurie A.

    1992-01-01

    Five years after graduation, responses from 519 nonvocational and 212 vocational Idaho graduates and 317 of their employers found no differences in satisfaction with curriculum; 40% nonvocational and 33% vocational jobs not related to high school plans; 89% nonvocational, 80% vocational concentrators satisfied with jobs; and no significant…

  7. Niacin alters the ruminal microbial composition of cattle under high-concentrate condition

    Directory of Open Access Journals (Sweden)

    Dan Luo

    2017-06-01

    Full Text Available To understand the effects of niacin on the ruminal microbial ecology of cattle under high-concentrate diet condition, Illumina MiSeq sequencing technology was used. Three cattle with rumen cannula were used in a 3 × 3 Latin-square design trial. Three diets were fed to these cattle during 3 periods for 3 days, respectively: high-forage diet (HF; forage-to-concentrate ratio = 80:20, high-concentrate diet (HC; forage-to-concentrate ratio = 20:80, and HC supplemented with 800 mg/kg niacin (HCN. Ruminal pH was measured before feeding and every 2 h after initiating feeding. Ruminal fluid was sampled at the end of each period for microbial DNA extraction. Overall, our findings revealed that subacute ruminal acidosis (SARA was induced and the α-diversity of ruminal bacterial community decreased in the cattle of HC group. Adding niacin in HC could relieve the symptoms of SARA in the cattle but the ruminal pH value and the Shannon index of ruminal bacterial community of HCN group were still lower than those of HF group. Whatever the diet was, the ruminal bacterial community of cattle was dominated by Bacteroidetes, Firmicutes and Proteobacteria. High-concentrate diet significantly increased the abundance of Prevotella, and decreased the abundance of Paraprevotella, Sporobacter, Ruminococcus and Treponema than HF. Compared with HC, HCN had a trend to decrease the percentage of Prevotella, and to increase the abundance of Succiniclasticum, Acetivibrio and Treponema. Increasing concentrate ratio could decrease ruminal pH value, and change the ruminal microbial composition. Adding niacin in HC could increase the ruminal pH value, alter the ruminal microbial composition.

  8. 509-45-1: A C. annuum Pepper germplasm containing high concentrations of capsinoids

    Science.gov (United States)

    This publication reports the public release of pepper (Capsicum annuum) germplasm ‘509-45-1’. Pepper germplasm 509-45-1 is a small-fruited, non-pungent single plant selection from PI 645509. Fruit of ‘509-45-1’ contain high concentrations of capsinoids [capsiate ((4-hydroxy-3-methoxybenzyl (E)-8...

  9. The preparation and ethanol fermentation of high-concentration sugars from steam-explosion corn stover.

    Science.gov (United States)

    Xie, Hui; Wang, Fengqin; Yin, Shuangyao; Ren, Tianbao; Song, Andong

    2015-05-01

    In the field of biofuel ethanol, high-concentration- reducing sugars made from cellulosic materials lay the foundation for high-concentration ethanol fermentation. In this study, corn stover was pre-treated in a process combining chemical methods and steam explosion; the cellulosic hydrolyzed sugars obtained by fed-batch saccharification were then used as the carbon source for high-concentration ethanol fermentation. Saccharomyces cerevisiae 1308, Angel yeast, and Issatchenkia orientalis were shake-cultured with Pachysolen tannophilus P-01 for fermentation. Results implied that the ethanol yields from the three types of mixed strains were 4.85 g/100 mL, 4.57 g/100 mL, and 5.02 g/100 mL (separately) at yield rates of 91.6, 89.3, and 92.2%, respectively. Therefore, it was inferred that shock-fermentation using mixed strains achieved a higher ethanol yield at a greater rate in a shorter fermentation period. This study provided a theoretical basis and technical guidance for the fermentation of industrial high-concentrated cellulosic ethanol.

  10. Low-Frequency Dielectric Dispersion of Highly Concentrated Spherical Particles in an Electrolyte Solution

    Institute of Scientific and Technical Information of China (English)

    倪福生; 顾国庆; 陈康民

    2002-01-01

    We deal with the problem of calculating the effective dielectric dispersion and electrical conductivity of colloidaldispersions. A comparison of the theoretical calculation of first principles with the experimental data of Schwanshows that our technique proposed here is no longer restricted to dilute solutions and is very effective for studyingthe dielectric properties of colloids with highly concentrated charged spherical particles in an electrolyte solution.

  11. Simplified yet highly accurate enzyme kinetics for cases of low substrate concentrations

    NARCIS (Netherlands)

    Härdin, H.M.; Zagaris, A.; Krab, K.; Westerhoff, H.V.

    2009-01-01

    Much of enzyme kinetics builds on simplifications enabled by the quasi-steady-state approximation and is highly useful when the concentration of the enzyme is much lower than that of its substrate. However, in vivo, this condition is often violated. In the present study, we show that, under conditio

  12. Continuum modeling of hydrodynamic particle–particle interactions in microfluidic high-concentration suspensions

    DEFF Research Database (Denmark)

    Ley, Mikkel Wennemoes Hvitfeld; Bruus, Henrik

    2016-01-01

    A continuum model is established for numerical studies of hydrodynamic particle–particle interactions in microfluidic high-concentration suspensions. A suspension of microparticles placed in a microfluidic channel and influenced by an external force, is described by a continuous particle...

  13. Analytical Study of High Concentration PCB Paint at the Heavy Water Components Test Reactor

    Energy Technology Data Exchange (ETDEWEB)

    Lowry, N.J.

    1998-10-21

    This report provides results of an analytical study of high concentration PCB paint in a shutdown nuclear test reactor located at the US Department of Energy's Savannah River Site (SRS). The study was designed to obtain data relevant for an evaluation of potential hazards associated with the use of and exposure to such paints.

  14. Decadal and seasonal trends of nutrient concentration and export from highly managed coastal catchments.

    Science.gov (United States)

    Wan, Yongshan; Wan, Lei; Li, Yuncong; Doering, Peter

    2017-05-15

    Understanding anthropogenic and hydro-climatic influences on nutrient concentrations and export from highly managed catchments often necessitates trend detection using long-term monitoring data. This study analyzed the temporal trend (1979-2014) of total nitrogen (TN) and total phosphorus (TP) concentrations and export from four adjacent coastal basins in south Florida where land and water resources are highly managed through an intricate canal network. The method of integrated seasonal-trend decomposition using LOESS (LOcally weighted regrESSion) was employed for trend detection. The results indicated that long-term trends in TN and TP concentrations (increasing/decreasing) varied with basins and nutrient species, reflecting the influence of basin specific land and water management practices. These long-term trends were intervened by short-term highs driven by high rainfall and discharges and lows associated with regional droughts. Seasonal variations in TP were more apparent than for TN. Nutrient export exhibited a chemostatic behavior for TN from all the basins, largely due to the biogenic nature of organic N associated with the ubiquity of organic materials in the managed canal network. Varying degrees of chemodynamic export was present for TP, reflecting complex biogeochemical responses to the legacy of long-term fertilization, low soil P holding capacity, and intensive stormwater management. The anthropogenic and hydro-climatic influences on nutrient concentration and export behavior had great implications in nutrient loading abatement strategies for aquatic ecosystem restoration of the downstream receiving waterbody. Published by Elsevier Ltd.

  15. Interculturalism in Italian Primary Schools with a High Concentration of Immigrant Students

    Science.gov (United States)

    Catarci, Marco

    2013-01-01

    The present article focuses on quantitative research carried out on a statistically representative sample of Italian primary schools with a high concentration of immigrant students. Research data show that schools with a higher number of immigrant students offer a greater number and a wider variety of intercultural initiatives. The presence of…

  16. Production of ethanol from wheat straw by pretreatment and fermentation at high dry matter concentrations

    NARCIS (Netherlands)

    Groenestijn, J.W. van; Slomp, R.S.

    2011-01-01

    High concentrations of substrate and product are important for the economy of second-generation bioethanol production. By a dilute acid thermal pretreatment of large pieces of relatively dry wheat straw using a novel rapid heating method, followed by fed-batch preliquefaction with hydrolytic enzymes

  17. Transient Dynamics of Fluoride-Based High Concentration Erbium/Cerium Co-Doped Fiber Amplifier

    Institute of Scientific and Technical Information of China (English)

    S. S-H. Yam; Y. Akasaka; Y. Kubota; R. Huang; D. L. Harris; J. Pan

    2003-01-01

    We designed and evaluated a fluoride-based high concentration erbium/ cerium co-doped fiber amplifier. It is suitable for Metropolitan Area Networks due to faster transient, flatter (unfiltered) gain, smaller footprint and gain excursion than its silica-based counterpart.

  18. Simplified yet highly accurate enzyme kinetics for cases of low substrate concentrations

    NARCIS (Netherlands)

    Härdin, H.M.; Zagaris, A.; Krab, K.; Westerhoff, H.V.

    2009-01-01

    Much of enzyme kinetics builds on simplifications enabled by the quasi-steady-state approximation and is highly useful when the concentration of the enzyme is much lower than that of its substrate. However, in vivo, this condition is often violated. In the present study, we show that, under

  19. Coherence and anticoherence resonance in high-concentration erbium-doped fiber laser

    OpenAIRE

    Sergeyev, Sergey; O'Mahoney, Kieran; Popov, Sergei; Friberg, Ari T.

    2010-01-01

    We report an experimental study of low-frequency (~10 kHz) self-pulsing of the output intensity in a high- concentration erbium-doped fiber laser. We suggest that the fast intensity fluctuations caused by multimode and polarization instabilities play the role of an external noise source, leading to low-frequency auto-oscillations through a coherence resonance scenario.

  20. Low powdered activated carbon concentrations to improve MBR sludge filterability at high salinity and low temperature

    NARCIS (Netherlands)

    Remy, M.J.J.; Temmink, B.G.; Brink, van den P.; Rulkens, W.H.

    2011-01-01

    Previous research has demonstrated that powdered activated carbon (PAC), when applied at very low dosages and long SRTs, reduces membrane fouling in membrane bioreactor (MBRs). This effect was related to stronger flocs which are less sensitive to shear. Low temperature and high salt concentration ar

  1. Mapping the Life Satisfaction of Adolescents in Hong Kong Secondary Schools with High Ethnic Concentration

    Science.gov (United States)

    Yuen, Yuet Mui Celeste; Lee, Moosung

    2016-01-01

    The present study aims to map the life satisfaction of adolescents from ethnic minority/immigrant backgrounds in schools with high concentrations of co-ethnic peers by comparing them with their mainstream counterparts in Hong Kong. The life satisfaction of 1,522 students was measured by the validated Multidimensional Students' Life Satisfaction…

  2. Red blood cell ATP/ADP & nitric oxide: The best vasodilators in diabetic patients

    Directory of Open Access Journals (Sweden)

    Bakhtiari Nuredin

    2012-08-01

    Full Text Available Abstract Background Diabetes mellitus is a group of metabolic diseases characterized by high blood sugar (glucose levels that result from defects in insulin secretion, or action, or both. Inspired by previous report the release of ATP from RBCs, which may participate in vessel dilation by stimulating NO production in the endothelium through purinergic receptor signaling and so, the aim of this study is to clearly determined relationship between RBC ATP/ADP ratio with nitric oxide. Methods The ATP/ADP ratio of erythrocytes among four groups of normal individuals (young & middle age, athletes’ subjects and diabetic patients were compared and the relationship between ATP/ADP ratio and NO level of plasma was determined with AVOVA test and bioluminescence method. Results ATP/ADP level in four groups normal (young & middle age, athletes, diabetes] are measured and analyzed with ANOVA test that show a significant difference between groups (P-value Conclusion In this study, a positive relationship between RBC ATP/ADP ratio and NO was found. Based on the obtained result, higher RBC ATP/ADP content may control the ratio of plasma NO in different individuals, also this results show that ATP can activate endothelial cells in NO production and is a main factor in releasing of NO from endothelial cells.

  3. Enhancing ATP-based bacteria and biofilm detection by enzymatic pyrophosphate regeneration.

    Science.gov (United States)

    Lee, Hui-Ju; Ho, Ming-Rong; Bhuwan, Manish; Hsu, Ching-Yi; Huang, Meng-Shun; Peng, Hwei-Ling; Chang, Hwan-You

    2010-04-15

    The manufacturing processes of many electronic and medical products demand the use of high-quality water. Hence the water supply systems for these processes are required to be examined regularly for the presence of microorganisms and microbial biofilms. Among commonly used bacteria detection approaches, the ATP luminescence assay is a rapid, sensitive, and easy to perform method. The aim of this study is to investigate whether ATP regeneration from inorganic pyrophosphate, a product of the ATP luminescence assay, can stabilize the bioluminescence signals in ATP detection. ADPglc pyrophosphorylase (AGPPase), which catalyzes the synthesis of ATP from PP(i) in the presence of ADPglc, was selected because the system yields much lower luminescence background than the commercially available ATP sulfurylase/adenosine 5'-phosphosulfate (APS) system which was broadly used in pyrosequencing technology. The AGPPase-based assay could be used to measure both PP(i) and ATP quantitatively and shows 1.5- to 4.0-fold slight increases in a 10-min assay. The method could also be used to stabilize the luminescence signals in detection of Escherichia coli, Pseudomonas aeruginosa, and Bacillus cereus in either broth or biofilm. These findings suggest that the AGPPase-based ATP regeneration system will find many practical applications such as detection of bacterial biofilm in water pipelines.

  4. Cell apoptosis and expression of ATP7A in brain of Atp7btx-J mice%Atp7btx-J小鼠脑组织细胞凋亡及ATP7A表达研究

    Institute of Scientific and Technical Information of China (English)

    胡璟; 焦先婷; 刘晓青; 余晓刚; 何振娟; 张拥军

    2013-01-01

    目的 通过对Atp7btx-J小鼠脑组织不同部位细胞凋亡的分析,钙、铜含量的测定,以及铜转运ATP酶ATP7A表达的研究,初步探讨肝豆状核变性致神经系统损伤的作用机制.方法 分离20周龄Atp7btx-J小鼠大脑皮层、小脑、基底神经节及海马区脑组织,Hoechst染色后分析细胞凋亡情况,电感耦合等离子体质谱法测定钙、铜含量,Real-Time PCR检测不同部位ATP7A mRNA 的表达量.结果 与野生型小鼠相比,纯合型小鼠小脑及基底神经节区细胞凋亡最为显著(P<0.05),小脑、基底神经节及海马区的铜和钙含量明显升高(P<0.01),脑组织不同部位ATP7A mRNA表达下调,其中基底神经节和小脑部位ATP7A mRNA表达的差异有统计学意义(P<0.05).结论 肝豆状核变性致神经系统损伤是多因素共同作用的结果,铜在脑内特殊部位的异常蓄积是始动因素,钙介导的细胞凋亡是导致神经系统损伤的重要因素,ATP7A在脑内不能代偿性地协助排出大量蓄积的铜可能加剧了神经系统的损伤.%Objective To explore the mechanism of nervous system injury in Wilson's disease by analysis of cell apoptosis,determination of concentrations of Ca and Cu and detection of expression of ATP7A in different parts of brain tissues of Atp7btx-J mice.Methods The tissues of cerebral cortex,cerebellum,basal ganglia and hippocampus of Atp7btx-J mice aged 20 weeks were isolated,the cell apoptosis was analyzed after Hoechst staining,the concentrations of Ca and Cu were determined by inductively coupled plasma mass spectrometry,and the expression of ATP7A mRNA in different parts was detected by Real-Time PCR.Results Compared with wild type mice,the cell apoptosis in cerebellum and basal ganglia of homozygous mice was most significant (P < 0.05).The concentrations of Ca and Cu in cerebellum,basal ganglia and hippocampus of homozygous mice were significantly higher than those of wild type mice (P < 0.01).The

  5. Consolidated bioprocessing of highly concentrated Jerusalem artichoke tubers for simultaneous saccharification and ethanol fermentation.

    Science.gov (United States)

    Guo, Lihao; Zhang, Jian; Hu, Fengxian; Dy Ryu, Dewey; Bao, Jie

    2013-10-01

    Consolidated bioprocessing (CBP) of Jerusalem artichoke tuber (Jat) for ethanol production is one of the most promising options for an alternate biofuel technology development. The technical barriers include the weak saccharolytic enzyme (inulinase) activity of the fermentation strain, and the well mixing of the high viscous fermentation slurry at the highly concentrated Jat loading. In this study, Saccharomyces cerevisiae DQ1 was found to produce relatively large amount of inulinase for hydrolysis of inulin in Jat, and the helical ribbon stirring bioreactor used provided well mixing performance under the high Jat loading. Even a highly concentrated Jat loading up to 35% (w/w) in the helical ribbon bioreactor for CBP was allowed. The results obtained from this study have demonstrated a feasibility of developing a CBP process technology in the helical ribbon bioreactor for ethanol production at a high yield 128.7 g/L and the theoretical yield 73.5%, respectively. This level of ethanol yield from Jat is relatively higher than others reported so far. The results of this study could provide a practical CBP process technology in the helical ribbon bioreactor for economically sustainable alternate biofuel production using highly concentrated inulin containing biomass feedstock such as Jat, at least 35%. Copyright © 2013 Wiley Periodicals, Inc.

  6. KCl -Permeabilized Pancreatic Islets: An Experimental Model to Explore the Messenger Role of ATP in the Mechanism of Insulin Secretion.

    Directory of Open Access Journals (Sweden)

    Javier Pizarro-Delgado

    Full Text Available Our previous work has demonstrated that islet depolarization with KCl opens connexin36 hemichannels in β-cells of mouse pancreatic islets allowing the exchange of small metabolites with the extracellular medium. In this study, the opening of these hemichannels has been further characterized in rat islets and INS-1 cells. Taking advantage of hemicannels'opening, the uptake of extracellular ATP and its effect on insulin release were investigated. 70 mM KCl stimulated light emission by luciferin in dispersed rat islets cells transduced with the fire-fly luciferase gene: it was suppressed by 20 mM glucose and 50 μM mefloquine, a specific connexin36 inhibitor. Extracellular ATP was taken up or released by islets depolarized with 70 mM KCl at 5 mM glucose, depending on the external ATP concentration. 1 mM ATP restored the loss of ATP induced by the depolarization itself. ATP concentrations above 5 mM increased islet ATP content and the ATP/ADP ratio. No ATP uptake occurred in non-depolarized or KCl-depolarized islets simultaneously incubated with 50 μM mefloquine or 20 mM glucose. Extracellular ATP potentiated the secretory response induced by 70 mM KCl at 5 mM glucose in perifused rat islets: 5 mM ATP triggered a second phase of insulin release after the initial peak triggered by KCl-depolarization itself; at 10 mM, it increased both the initial, KCl-dependent, peak and stimulated a greater second phase of secretion than at 5 mM. These stimulatory effects of extracellular ATP were almost completely suppressed by 50 μM mefloquine. The magnitude of the second phase of insulin release due to 5 mM extracellular ATP was decreased by addition of 5 mM ADP (extracellular ATP/ADP ratio = 1. ATP acts independently of KATP channels closure and its intracellular concentration and its ATP/ADP ratio seems to regulate the magnitude of both the first (triggering and second (amplifying phases of glucose-induced insulin secretion.

  7. Concentration-Induced Association in a Protein System Caused by a Highly Directional Patch Attraction.

    Science.gov (United States)

    Li, Weimin; Persson, Björn A; Lund, Mikael; Bergenholtz, Johan; Zackrisson Oskolkova, Malin

    2016-09-01

    Self-association of the protein lactoferrin is studied in solution using small-angle X-ray scattering techniques. Effective static structure factors have been shown to exhibit either a monotonic or a nonmonotonic dependence on protein concentration in the small wavevector limit, depending on salt concentration. The behavior correlates with a nonmonotonic dependence of the second virial coefficient on salt concentration, such that a maximum appears in the structure factor at a low protein concentration when the second virial coefficient is negative and close to a minimum. The results are interpreted in terms of an integral equation theory with explicit dimers, formulated by Wertheim, which provides a consistent framework able to explain the behavior in terms of a monomer-dimer equilibrium that appears because of a highly directional patch attraction. Short attraction ranges preclude trimer formation, which explains why the protein system behaves as if it were subject to a concentration-dependent isotropic protein-protein attraction. Superimposing an isotropic interaction, comprising screened Coulomb repulsion and van der Waals attraction, on the patch attraction allows for a semiquantitative modeling of the complete transition pathway from monomers in the dilute limit to monomer-dimer systems at somewhat higher protein concentrations.

  8. The molecular interaction of a protein in highly concentrated solution investigated by Raman spectroscopy.

    Science.gov (United States)

    Ota, Chikashi; Noguchi, Shintaro; Tsumoto, Kouhei

    2015-04-01

    We used Raman spectroscopy to investigate the structure and interactions of lysozyme molecules in solution over a wide range of concentrations (2.5-300 mg ml(-1)). No changes in the amide-I band were observed as the concentration was increased, but the width of the Trp band at 1555 cm(-1) and the ratios of the intensities of the Tyr bands at 856 and 837 cm(-1), the Trp bands at 870 and 877 cm(-1), and the bands at 2940 (CH stretching) and 3420 cm(-1) (OH stretching) changed as the concentration was changed. These results reveal that although the distance between lysozyme molecules changed by more than an order of magnitude over the tested concentration range, the secondary structure of the protein did not change. The changes in the molecular interactions occurred in a stepwise process as the order of magnitude of the distance between molecules changed. These results suggest that Raman bands can be used as markers to investigate the behavior of high-concentration solutions of proteins and that the use of Raman spectroscopy will lead to progress in our understanding not only of the basic science of protein behavior under concentrated (i.e., crowded) conditions but also of practical processes involving proteins, such as in the field of biopharmaceuticals. © 2014 Wiley Periodicals, Inc.

  9. Rapid estimation of concentration of aromatic classes in middistillate fuels by high-performance liquid chromatography

    Science.gov (United States)

    Otterson, D. A.; Seng, G. T.

    1985-01-01

    An high performance liquid chromatography (HPLC) method to estimate four aromatic classes in middistillate fuels is presented. Average refractive indices are used in a correlation to obtain the concentrations of each of the aromatic classes from HPLC data. The aromatic class concentrations can be obtained in about 15 min when the concentration of the aromatic group is known. Seven fuels with a wide range of compositions were used to test the method. Relative errors in the concentration of the two major aromatic classes were not over 10 percent. Absolute errors of the minor classes were all less than 0.3 percent. The data show that errors in group-type analyses using sulfuric acid derived standards are greater for fuels containing high concentrations of polycyclic aromatics. Corrections are based on the change in refractive index of the aromatic fraction which can occur when sulfuric acid and the fuel react. These corrections improved both the precision and the accuracy of the group-type results.

  10. Effects of exposure to high concentrations of waterborne Tl on K and Tl concentrations in Chironomus riparius larvae.

    Science.gov (United States)

    Belowitz, Ryan; Leonard, Erin M; O'Donnell, Michael J

    2014-11-01

    Thallium (Tl) is a non-essential metal which is released into the environment primarily as the result of anthropogenic activities such as fossil fuel burning and smelting of ores. The ionic radius of monovalent Tl⁺ is similar to that of K⁺ and Tl⁺ may thus interfere with K⁺-dependent processes. We determined that the acute (48 h) lethal concentration where 50% of the organisms do not survive (LC₅₀) of Tl for 4th instar Chironomus riparius larvae was 723 μmol L⁻¹. Accumulation of Tl by the whole animal was saturable, with a maximum accumulation (Jmax) of 4637 μmol kg⁻¹ wet mass, and K(D) of 670 μmol Tl l⁻¹. Tl accumulation by the gut appeared saturable at the lowest four Tl concentrations, with a Jmax of 2560 μmol kg⁻¹ wet mass and a K(D) of 54.5 μmol Tl l⁻¹. The saturable accumulation at the gut may be indicative of a limited capacity for intracellular detoxification, such as storage in lysosomes or complexation with metal-binding proteins. Tl accumulation by the hemolymph was found to be linear and Tl concentrations in the hemolymph were ~75% of the exposure concentration at Tl exposures >26.9 μmol L⁻¹. There was not a significant decrease in whole animal, gut or hemolymph K during exposure to waterborne Tl at any of the concentrations tested (up to 1500 μmol L⁻¹). The avoidance of hypokalemia by C. riparius larvae may contribute to survival during acute waterborne exposures to Tl. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. Effects of prior acute exercise on circulating cytokine concentration responses to a high-fat meal.

    Science.gov (United States)

    Brandauer, Josef; Landers-Ramos, Rian Q; Jenkins, Nathan T; Spangenburg, Espen E; Hagberg, James M; Prior, Steven J

    2013-08-01

    High-fat meal consumption alters the circulating cytokine profile and contributes to cardiometabolic diseases. A prior bout of exercise can ameliorate the triglyceride response to a high-fat meal, but the interactive effects of exercise and high-fat meals on cytokines that mediate cardiometabolic risk are not fully understood. We investigated the effects of prior exercise on the responses of circulating tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-8, leptin, retinol-binding protein 4 (RBP4), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), placental growth factor (PlGF), and soluble fms-like tyrosine kinase-1 (sFlt-1) to a high-fat meal. Ten healthy men were studied before and 4 h after ingestion of a high-fat meal either with or without ∼50 min of endurance exercise at 70% of VO2 max on the preceding day. In response to the high-fat meal, lower leptin and higher VEGF, bFGF, IL-6, and IL-8 concentrations were evident (P exercise (P exercise and the high-fat meal on sFlt-1 (P exercise and 218% with prior exercise (P exercise does not affect all high-fat meal-induced changes in circulating cytokines, but does affect fasting or postprandial concentrations of IL-6, leptin, and sFlt-1. These data may reflect a salutary effect of prior exercise on metabolic responses to a high-fat meal.

  12. UTP-induced ATP release is a fine-tuned signalling pathway in osteocytes

    DEFF Research Database (Denmark)

    Kringelbach, Tina M.; Aslan, Derya; Novak, Ivana

    2014-01-01

    that ATP is released load-dependently from osteocytes from the onset of mechanical stimulation. Therefore, the aim of the present study was to investigate whether and how ATP release can be evoked in osteocytes via purinergic receptor activation. ATP release was quantified by real-time determination using...... responses to P2 receptor agonists. These investigations demonstrated that MLO-Y4 osteocytes express functional P2Y2, P2Y4, P2Y12 and P2Y13 receptors in addition to the previously reported P2X receptors. Further, we found that osteocytes respond to nucleotides such as ATP, UTP and ADP by increasing...... the intracellular calcium concentration and that they release ATP dose-dependently upon stimulation with 1-10 μM UTP. In addition to this, osteocytes release large amounts of ATP upon cell rupture, which might also be a source for other nucleotides, such as UTP. These findings indicate that mechanically induced ATP...

  13. Plasma PCSK9 concentrations during an oral fat load and after short term high-fat, high-fat high-protein and high-fructose diets

    Directory of Open Access Journals (Sweden)

    Cariou Bertrand

    2013-01-01

    Full Text Available Abstract Background PCSK9 (Proprotein Convertase Subtilisin Kexin type 9 is a circulating protein that promotes hypercholesterolemia by decreasing hepatic LDL receptor protein. Under non interventional conditions, its expression is driven by sterol response element binding protein 2 (SREBP2 and follows a diurnal rhythm synchronous with cholesterol synthesis. Plasma PCSK9 is associated to LDL-C and to a lesser extent plasma triglycerides and insulin resistance. We aimed to verify the effect on plasma PCSK9 concentrations of dietary interventions that affect these parameters. Methods We performed nutritional interventions in young healthy male volunteers and offspring of type 2 diabetic (OffT2D patients that are more prone to develop insulin resistance, including: i acute post-prandial hyperlipidemic challenge (n=10, ii 4 days of high-fat (HF or high-fat/high-protein (HFHP (n=10, iii 7 (HFruc1, n=16 or 6 (HFruc2, n=9 days of hypercaloric high-fructose diets. An acute oral fat load was also performed in two patients bearing the R104C-V114A loss-of-function (LOF PCSK9 mutation. Plasma PCSK9 concentrations were measured by ELISA. For the HFruc1 study, intrahepatocellular (IHCL and intramyocellular lipids were measured by 1H magnetic resonance spectroscopy. Hepatic and whole-body insulin sensitivity was assessed with a two-step hyperinsulinemic-euglycemic clamp (0.3 and 1.0 mU.kg-1.min-1. Findings HF and HFHP short-term diets, as well as an acute hyperlipidemic oral load, did not significantly change PCSK9 concentrations. In addition, post-prandial plasma triglyceride excursion was not altered in two carriers of PCSK9 LOF mutation compared with non carriers. In contrast, hypercaloric 7-day HFruc1 diet increased plasma PCSK9 concentrations by 28% (p=0.05 in healthy volunteers and by 34% (p=0.001 in OffT2D patients. In another independent study, 6-day HFruc2 diet increased plasma PCSK9 levels by 93% (p Conclusions Plasma PCSK9 concentrations vary

  14. Design and Optimization of Fresnel Lens for High Concentration Photovoltaic System

    Directory of Open Access Journals (Sweden)

    Lei Jing

    2014-01-01

    Full Text Available A practical optimization design is proposed, in which the solar direct light spectrum and multijunction cell response range are taken into account in combination, particularly for the Fresnel concentrators with a high concentration and a small aspect ratio. In addition, the change of refractive index due to temperature variation in outdoor operation conditions is also considered in the design stage. The calculation results show that this novel Fresnel lens achieves an enhancement of energy efficiency of about 10% compared with conventional Fresnel lens for a given solar spectrum, solar cell response, and corrected sunshine hours of different ambient temperature intervals.

  15. Optimal Dearsenification Parameters of Gold Sulfide Concentrate with a High As Content

    Institute of Scientific and Technical Information of China (English)

    Yaozhong LAN; Rose W. Smith

    2005-01-01

    The gold sulfide concentrate with a high As content in Liangshan District, Sichuan Province, China, is a potentially important resource. This paper describes experiments of dearsenification of gold concentrate in a weakly reduced atmosphere in a rotary pipe furnace. The results showed that the optimal parameters were a temperature range of 650~700℃, 15%~16% CO2 of gas and a reaction time of 30~40 min. The removal rate of arsenic and sulfur was over 95% and 25%~28%, respectively. With further oxidization and roasting, residue sulfur in the roasted materials was dropped to below 4%, and the cyanide leaching recovery of gold was over 92%.

  16. Enhanced Cycling Stability of Rechargeable Li-O2 Batteries Using High Concentration Electrolytes

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Bin; Xu, Wu; Yan, Pengfei; Sun, Xiuliang; Bowden, Mark E.; Read, Jeffrey; Qian, Jiangfeng; Mei, Donghai; Wang, Chong M.; Zhang, Jiguang

    2016-01-26

    The electrolyte stability against reactive reduced-oxygen species is crucial for the development of rechargeable Li-O2 batteries. In this work, we systematically investigated the effect of lithium salt concentration in 1,2-dimethoxyethane (DME)-based electrolytes on the cycling stability of Li-O2 batteries. Cells with high concentration electrolyte illustrate largely enhanced cycling stability under both the full discharge/charge (2.0-4.5 V vs. Li/Li+) and the capacity limited (at 1,000 mAh g-1) conditions. These cells also exhibit much less reaction-residual on the charged air electrode surface, and much less corrosion to the Li metal anode. The density functional theory calculations are conducted on the molecular orbital energies of the electrolyte components and the Gibbs activation barriers for superoxide radical anion to attack DME solvent and Li+-(DME)n solvates. In a highly concentrated electrolyte, all DME molecules have been coordinated with salt and the C-H bond scission of a DME molecule becomes more difficult. Therefore, the decomposition of highly concentrated electrolyte in a Li-O2 battery can be mitigated and both air-cathodes and Li-metal anodes exhibits much better reversibility. As a results, the cyclability of Li-O2 can be largely improved.

  17. High nitrogen supply and carbohydrate content reduce fungal endophyte and alkaloid concentration in Lolium perenne.

    Science.gov (United States)

    Rasmussen, Susanne; Parsons, Anthony J; Bassett, Shalome; Christensen, Michael J; Hume, David E; Johnson, Linda J; Johnson, Richard D; Simpson, Wayne R; Stacke, Christina; Voisey, Christine R; Xue, Hong; Newman, Jonathan A

    2007-01-01

    The relationship between cool-season grasses and fungal endophytes is widely regarded as mutualistic, but there is growing uncertainty about whether changes in resource supply and environment benefit both organisms to a similar extent. Here, we infected two perennial ryegrass (Lolium perenne) cultivars (AberDove, Fennema) that differ in carbohydrate content with three strains of Neotyphodium lolii (AR1, AR37, common strain) that differ intrinsically in alkaloid profile. We grew endophyte-free and infected plants under high and low nitrogen (N) supply and used quantitative PCR (qPCR) to estimate endophyte concentrations in harvested leaf tissues. Endophyte concentration was reduced by 40% under high N supply, and by 50% in the higher sugar cultivar. These two effects were additive (together resulting in 75% reduction). Alkaloid production was also reduced under both increased N supply and high sugar cultivar, and for three of the four alkaloids quantified, concentrations were linearly related to endophyte concentration. The results stress the need for wider quantification of fungal endophytes in the grassland-foliar endophyte context, and have implications for how introducing new cultivars, novel endophytes or increasing N inputs affect the role of endophytes in grassland ecosystems.

  18. Hyperproduction of Alcohol Using Yeast Fermentation in Highly Concentrated Molasses Medium

    Institute of Scientific and Technical Information of China (English)

    顾燕松; 周政懋; 乔敏; 周全; 陈国强

    2001-01-01

    Cane molasses, a major byproduct in the sugar industry, is generally consumed for alcohol production. However, the alcohol production process needs to overcome three major challenges including increasing the productivity of alcohol fermentation, lowering the energy consumption for alcohol conversion and decreasing the environmental pollution caused by the alcoholic yeast fermentation process. To meet these challenges, a screening process was conducted using 13 high osmotic tolerant yeast strains. Among the strains, a Saccharomyces cerevisiae strain 1912 was found to produce high alcohol concentrations during fermentation with high starting molasses concentrations such as 50% (W/V) molasses. In the test, 13.6% (V/V) alcohol was produced in the molasses fermentation broth after 72 h of incubation with an initial Yunnan molasses concentration of 50% in a 5 L fermentor. 15.0% (V/V) alcohol was obtained after 48 h of fermentation in shaking flasks containing 30% (W/V) initial total sugar concentration in diluted molasses. The performance of this strain in the shaking flasks was successfully scaled up to a 5-L fermentor vessel. Strain 1912 seems to be a better alcohol producer than the currently used alcohol production strain 2. 1190.

  19. Aggregation of normal and sickle hemoglobin in high concentration phosphate buffer.

    Science.gov (United States)

    Chen, Kejing; Ballas, Samir K; Hantgan, Roy R; Kim-Shapiro, Daniel B

    2004-12-01

    Sickle cell disease is caused by a mutant form of hemoglobin, hemoglobin S, that polymerizes under hypoxic conditions. The extent and mechanism of polymerization are thus the subject of many studies of the pathophysiology of the disease and potential treatment strategies. To facilitate such studies, a model system using high concentration phosphate buffer (1.5 M-1.8 M) has been developed. To properly interpret results from studies using this model it is important to understand the similarities and differences in hemoglobin S polymerization in the model compared to polymerization under physiological conditions. In this article, we show that hemoglobin S and normal adult hemoglobin, hemoglobin A, aggregate in high concentration phosphate buffer even when the concentration of hemoglobin is below the solubility defined for polymerization. This phenomenon was not observed using 0.05 M phosphate buffer or in another model system we studied that uses dextran to enhance polymerization. We have used static light scattering, dynamic light scattering, and differential interference contrast microscopy to confirm aggregation of deoxygenated and oxygenated hemoglobins below their solubility and have shown that this aggregation is not observable using turbidity measurements, a common technique for assessing polymerization. We have also shown that the aggregation increases with increasing temperature in the range of 15 degrees -37 degrees C and that it increases as the concentration of phosphate increases. These studies contribute to the working knowledge of how to properly apply studies of hemoglobin S polymerization that are conducted using the high phosphate model.

  20. Quality evaluation of moluodan concentrated pill using high-performance liquid chromatography fingerprinting coupled with chemometrics.

    Science.gov (United States)

    Tao, Lingyan; Zhang, Qing; Wu, Yongjiang; Liu, Xuesong

    2016-12-01

    In this study, a fast and effective high-performance liquid chromatography method was developed to obtain a fingerprint chromatogram and quantitative analysis simultaneously of four indexes including gallic acid, chlorogenic acid, albiflorin and paeoniflorin of the traditional Chinese medicine Moluodan Concentrated Pill. The method was performed by using a Waters X-bridge C18 reversed phase column on an Agilent 1200S high-performance liquid chromatography system coupled with diode array detection. The mobile phase of the high-performance liquid chromatography method was composed of 20 mmol/L phosphate solution and acetonitrile with a 1 mL/min eluent velocity, under a detection temperature of 30°C and a UV detection wavelength of 254 nm. After the methodology validation, 16 batches of Moluodan Concentrated Pill were analyzed by this high-performance liquid chromatography method and both qualitative and quantitative evaluation results were achieved by similarity analysis, principal component analysis and hierarchical cluster analysis. The results of these three chemometrics were in good agreement and all indicated that batch 10 and batch 16 showed significant differences with the other 14 batches. This suggested that the developed high-performance liquid chromatography method could be applied in the quality evaluation of Moluodan Concentrated Pill.

  1. Vasopressin activates Akt/mTOR pathway in smooth muscle cells cultured in high glucose concentration

    Energy Technology Data Exchange (ETDEWEB)

    Montes, Daniela K.; Brenet, Marianne; Muñoz, Vanessa C.; Burgos, Patricia V.; Villanueva, Carolina I. [Department of Physiology, Universidad Austral de Chile, Valdivia 509-9200 (Chile); Figueroa, Carlos D. [Department of Anatomy, Histology and Pathology, Universidad Austral de Chile, Valdivia 509-9200 (Chile); González, Carlos B., E-mail: cbgonzal@uach.cl [Department of Physiology, Universidad Austral de Chile, Valdivia 509-9200 (Chile); Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, TX 77555 (United States)

    2013-11-29

    Highlights: •AVP induces mTOR phosphorylation in A-10 cells cultured in high glucose concentration. •The mTOR phosphorylation is mediated by the PI3K/Akt pathway activation. •The AVP-induced mTOR phosphorylation inhibited autophagy and stimulated cell proliferation. -- Abstract: Mammalian target of rapamycin (mTOR) complex is a key regulator of autophagy, cell growth and proliferation. Here, we studied the effects of arginine vasopressin (AVP) on mTOR activation in vascular smooth muscle cells cultured in high glucose concentration. AVP induced the mTOR phosphorylation in A-10 cells grown in high glucose, in contrast to cells cultured in normal glucose; wherein, only basal phosphorylation was observed. The AVP-induced mTOR phosphorylation was inhibited by a PI3K inhibitor. Moreover, the AVP-induced mTOR activation inhibited autophagy and increased thymidine incorporation in cells grown in high glucose. This increase was abolished by rapamycin which inhibits the mTORC1 complex formation. Our results suggest that AVP stimulates mTOR phosphorylation by activating the PI3K/Akt signaling pathway and, subsequently, inhibits autophagy and raises cell proliferation in A-10 cells maintained in high glucose concentration.

  2. The Effects of High Concentrations of Vitamin C on Cancer Cells

    Directory of Open Access Journals (Sweden)

    Seyeon Park

    2013-09-01

    Full Text Available The effect of high doses of vitamin C for the treatment of cancer has been controversial. Our previous studies, and studies by others, have reported that vitamin C at concentrations of 0.25–1.0 mM induced a dose- and time-dependent inhibition of proliferation in acute myeloid leukemia (AML cell lines and in leukemic cells from peripheral blood specimens obtained from patients with AML. Treatment of cells with high doses of vitamin C resulted in an immediate increase in intracellular total glutathione content and glutathione-S transferase activity that was accompanied by the uptake of cysteine. These results suggest a new role for high concentrations of vitamin C in modulation of intracellular sulfur containing compounds, such as glutathione and cysteine. This review, discussing biochemical pharmacologic studies, including pharmacogenomic and pharmacoproteomic studies, presents the different pharmacological effects of vitamin C currently under investigation.

  3. Influence of a High-Pressure Comminution Technology on Concentrate Yields in Copper Ore Flotation Processes

    Directory of Open Access Journals (Sweden)

    Saramak D.

    2014-10-01

    Full Text Available The article concerns the issues of flotation process effectiveness in relationship to the operating conditions of a high-pressure comminution process course. Experimental programme covering a flotation laboratory batch tests was a verification technique of a high-pressure crushing operations course. The most favorable values of flotation concentrate weight recoveries were obtained for the pressing force 6 kN and 4% of the feed moisture. It was also determined the model of the concentrate weight recovery as a function of pressing force in the press and feed moisture content. This model was the basis for the optimization of effects of copper ore flotation processes preceded in high-pressure crushing operation in roller presses.

  4. Fabrication of High Contrast Gratings for the Spectrum Splitting Dispersive Element in a Concentrated Photovoltaic System.

    Science.gov (United States)

    Yao, Yuhan; Liu, He; Wu, Wei

    2015-07-18

    High contrast gratings are designed and fabricated and its application is proposed in a parallel spectrum splitting dispersive element that can improve the solar conversion efficiency of a concentrated photovoltaic system. The proposed system will also lower the solar cell cost in the concentrated photovoltaic system by replacing the expensive tandem solar cells with the cost-effective single junction solar cells. The structures and the parameters of high contrast gratings for the dispersive elements were numerically optimized. The large-area fabrication of high contrast gratings was experimentally demonstrated using nanoimprint lithography and dry etching. The quality of grating material and the performance of the fabricated device were both experimentally characterized. By analyzing the measurement results, the possible side effects from the fabrication processes are discussed and several methods that have the potential to improve the fabrication processes are proposed, which can help to increase the optical efficiency of the fabricated devices.

  5. Spectral and Concentration Sensitivity of Multijunction Solar Cells at High Temperature: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Friedman, Daniel J.; Steiner, Myles A.; Perl, Emmett E.; Simon, John

    2017-06-14

    We model the performance of two-junction solar cells at very high temperatures of ~400 degrees C and beyond for applications such as hybrid PV/solar-thermal power production, and identify areas in which the design and performance characteristics behave significantly differently than at more conventional near-room-temperature operating conditions. We show that high-temperature operation reduces the sensitivity of the cell efficiency to spectral content, but increases the sensitivity to concentration, both of which have implications for energy yield in terrestrial PV applications. For other high-temperature applications such as near-sun space missions, our findings indicate that concentration may be a useful tool to enhance cell efficiency.

  6. Microstructural Changes in High-Protein Nutrition Bars Formulated with Extruded or Toasted Milk Protein Concentrate.

    Science.gov (United States)

    Banach, J C; Clark, S; Lamsal, B P

    2016-02-01

    Milk protein concentrates with more than 80% protein (that is, MPC80) are underutilized as the primary protein source in high-protein nutrition bars as they impart crumbliness and cause hardening during storage. High-protein nutrition bar texture changes are often associated with internal protein aggregations and macronutrient phase separation. These changes were investigated in model high-protein nutrition bars formulated with MPC80 and physically modified MPC80s. High-protein nutrition bars formulated with extruded MPC80s hardened slower than those formulated with toasted or unmodified MPC80. Extruded MPC80 had reduced free sulfhydryl group exposure, whereas measurable increases were seen in the toasted MPC80. High-protein nutrition bar textural performance may be related to the number of exposed free sulfhydryl groups in MPC80. Protein aggregations resulting from ingredient modification and high-protein nutrition bar storage were studied with sodium dodecyl sulfate polyacrylamide gel electrophoresis. Disulfide-based protein aggregations and changes in free sulfhydryl concentration were not consistently relatable to high-protein nutrition bar texture change. However, the high-protein nutrition bars formulated with extruded MPC80 were less prone to phase separations, as depicted by confocal laser scanning microscopy, and underwent less texture change during storage than those formulated with toasted or unmodified MPC80.

  7. Adaptation and acclimatization to formaldehyde in methylotrophs capable of high-concentration formaldehyde detoxification.

    Science.gov (United States)

    Chongcharoen, Rotsaman; Smith, Thomas J; Flint, Kenneth P; Dalton, Howard

    2005-08-01

    Formaldehyde is a highly toxic chemical common in industrial effluents, and it is also an intermediate in bacterial metabolism of one-carbon growth substrates, although its role as a bacterial growth substrate per se has not been extensively reported. This study investigated two highly formaldehyde-resistant formaldehyde utilizers, strains BIP and ROS1; the former strain has been used for industrial remediation of formaldehyde-containing effluents. The two strains were shown by means of 16S rRNA characterization to be closely related members of the genus Methylobacterium. Both strains were able to use formaldehyde, methanol and a range of multicarbon compounds as their principal growth substrate. Growth on formaldehyde was possible up to a concentration of at least 58 mM, and survival at up to 100 mM was possible after stepwise acclimatization by growth at increasing concentrations of formaldehyde. At such high concentrations of formaldehyde, the cultures underwent a period of formaldehyde removal without growth before the formaldehyde concentration fell below 60 mM, and growth could resume. Two-dimensional electrophoresis and MS characterization of formaldehyde-induced proteins in strain BIP revealed that the pathways of formaldehyde metabolism, and adaptations to methylotrophic growth, were very similar to those seen in the well-characterized methanol-utilizing methylotroph Methylobacterium extorquens AM1. Thus, it appears that many of the changes in protein expression that allow strain BIP to grow using high formaldehyde concentrations are associated with expression of the same enzymes used by M. extorquens AM1 to process formaldehyde as a metabolic intermediate during growth on methanol.

  8. Circulating adiponectin concentration and body composition are altered in response to high-intensity interval training.

    Science.gov (United States)

    Shing, Cecilia M; Webb, Jessica J; Driller, Matthew W; Williams, Andrew D; Fell, James W

    2013-08-01

    Adiponectin influences metabolic adaptations that would prove beneficial to endurance athletes, and yet to date there is little known about the response of adiponectin concentrations to exercise, and, in particular, the response of this hormone to training in an athlete population. This study aimed to determine the response of plasma adiponectin concentrations to acute exercise after 2 different training programs and to determine the influence of the training on body composition. Seven state-level representative rowers (age: 19 ± 1.2 years [mean ± SD], height: 1.77 ± 0.10 m, body mass: 74.0 ± 10.7 kg, VO2peak 62.1 ± 7.0 ml·kg·min) participated in the double-blind, randomized crossover investigation. Rowers performed an incremental graded exercise test before and after completing 4 weeks of high-intensity interval ergometer training and 4 weeks of traditional ergometer rowing training. Rowers' body composition was assessed at baseline and after each training program. Significant increases in plasma adiponectin concentration occurred in response to maximal exercise after completion of the high-intensity interval training (p = 0.016) but not after traditional ergometer rowing training (p = 0.69). The high-intensity interval training also resulted in significant increases in mean 4-minute power output (p = 0.002) and VO2peak (p = 0.05), and a decrease in body fat percentage (p = 0.022). Mean 4-minute power output, VO2peak, and body fat percentage were not significantly different after 4 weeks of traditional ergometer rowing training (p > 0.05). Four weeks of high-intensity interval training is associated with an increase in adiponectin concentration in response to maximal exercise and a reduction in body fat percentage. The potential for changes in adiponectin concentration to reflect positive training adaptations and athlete performance level should be further explored.

  9. Serum POP concentrations are highly predictive of inner blubber concentrations at two extremes of body condition in northern elephant seals.

    Science.gov (United States)

    Peterson, Michael G; Peterson, Sarah H; Debier, Cathy; Covaci, Adrian; Dirtu, Alin C; Malarvannan, Govindan; Crocker, Daniel E; Costa, Daniel P

    2016-11-01

    Long-lived, upper trophic level marine mammals are vulnerable to bioaccumulation of persistent organic pollutants (POPs). Internal tissues may accumulate and mobilize POP compounds at different rates related to the body condition of the animal and the chemical characteristics of individual POP compounds; however, collection of samples from multiple tissues is a major challenge to ecotoxicology studies of free-ranging marine mammals and the ability to predict POP concentrations in one tissue from another tissue remains rare. Northern elephant seals (Mirounga angustirostris) forage on mesopelagic fish and squid for months at a time in the northeastern Pacific Ocean, interspersed with two periods of fasting on land, which results in dramatic seasonal fluctuations in body condition. Using northern elephant seals, we examined commonly studied tissues in mammalian toxicology to describe relationships and determine predictive equations among tissues for a suite of POP compounds, including ΣDDTs, ΣPCBs, Σchlordanes, and ΣPBDEs. We collected paired blubber (inner and outer) and blood serum samples from adult female and male seals in 2012 and 2013 at Año Nuevo State Reserve (California, USA). For females (N = 24), we sampled the same seals before (late in molting fast) and after (early in breeding fast) their approximately seven month foraging trip. For males, we sampled different seals before (N = 14) and after (N = 15) their approximately four month foraging trip. We observed strong relationships among tissues for many, but not all compounds. Serum POP concentrations were strong predictors of inner blubber POP concentrations for both females and males, while serum was a more consistent predictor of outer blubber for males than females. The ability to estimate POP blubber concentrations from serum, or vice versa, has the potential to enhance toxicological assessment and physiological modeling. Furthermore, predictive equations may illuminate commonalities or

  10. Lidar detection of high concentrations of ozone and aerosol transported from northeastern Asia over Saga, Japan

    Science.gov (United States)

    Uchino, Osamu; Sakai, Tetsu; Izumi, Toshiharu; Nagai, Tomohiro; Morino, Isamu; Yamazaki, Akihiro; Deushi, Makoto; Yumimoto, Keiya; Maki, Takashi; Tanaka, Taichu Y.; Akaho, Taiga; Okumura, Hiroshi; Arai, Kohei; Nakatsuru, Takahiro; Matsunaga, Tsuneo; Yokota, Tatsuya

    2017-02-01

    To validate products of the Greenhouse gases Observing SATellite (GOSAT), we observed vertical profiles of aerosols, thin cirrus clouds, and tropospheric ozone with a mobile-lidar system that consisted of a two-wavelength (532 and 1064 nm) polarization lidar and a tropospheric ozone differential absorption lidar (DIAL). We used these lidars to make continuous measurements over Saga (33.24° N, 130.29° E) during 20-31 March 2015. High ozone and high aerosol concentrations were observed almost simultaneously in the altitude range 0.5-1.5 km from 03:00 to 20:00 Japan Standard Time (JST) on 22 March 2015. The maximum ozone volume mixing ratio was ˜ 110 ppbv. The maxima of the aerosol extinction coefficient and optical depth at 532 nm were 1.2 km-1 and 2.1, respectively. Backward trajectory analysis and the simulations by the Model of Aerosol Species IN the Global AtmospheRe (MASINGAR) mk-2 and the Meteorological Research Institute Chemistry-Climate Model, version 2 (MRI-CCM2), indicated that mineral dust particles from the Gobi Desert and an air mass with high ozone and aerosol (mainly sulfate) concentrations that originated from the North China Plain could have been transported over the measurement site within about 2 days. These high ozone and aerosol concentrations impacted surface air quality substantially in the afternoon of 22 March 2015. After some modifications of its physical and chemical parameters, MRI-CCM2 approximately reproduced the high ozone volume mixing ratio. MASINGAR mk-2 successfully predicted high aerosol concentrations, but the predicted peak aerosol optical thickness was about one-third of the observed value.

  11. Material and Optical Design Rules for High Performance Luminescent Solar Concentrators

    Science.gov (United States)

    Bronstein, Noah Dylan

    This dissertation will highlight a path to achieve high photovoltaic conversion efficiency in luminescent solar concentrators, devices which absorb sunlight with a luminescent dye and then re-emit it into a waveguide where it is ultimately collected by a photovoltaic cell. Luminescent concentrators have been studied for more than three decades as potential low-cost but not high efficiency photovoltaics. Astute application of the blackbody radiation law indicates that photonic design is necessary to achieve high efficiency: a reflective filter must be used to trap luminescence at all angles while allowing higher energy photons to pass through. In addition, recent advances in the synthesis of colloidal nanomaterials have created the possibility for lumophores with broad absorption spectra, narrow-bandwidth emission, high luminescence quantum yield, tunable Stokes shifts and tunable Stokes ratios. Together, these factors allow luminescent solar concentrators to achieve the optical characteristics necessary for high efficiency. We have fabricated and tested the first generation of these devices. Our experiments demonstrate that the application of carefully matched photonic mirrors and luminescent quantum dots can allow luminescent concentration factors to reach record values while maintaining high photon collection efficiency. Finally, the photonic mirror dramatically mitigates the negative impact of scattering in the waveguide, allowing efficient photon collection over distances much longer than the scattering length of the waveguide. After demonstrating the possibility for high performance, we theoretically explore the efficacy of luminescent concentrators with dielectric reflectors as the high-bandgap top-junctions in two-junction devices. Simple thermodynamic calculations indicate that this approach can be nearly as good as a traditional vertically stacked tandem. The major barriers to such a device are the optical design of narrow-bandwidth, angle

  12. Effect of Temperature on Chinese Rice Wine Brewing with High Concentration Presteamed Whole Sticky Rice

    Directory of Open Access Journals (Sweden)

    Dengfeng Liu

    2014-01-01

    Full Text Available Production of high quality Chinese rice wine largely depends on fermentation temperature. However, there is no report on the ethanol, sugars, and acids kinetics in the fermentation mash of Chinese rice wine treated at various temperatures. The effects of fermentation temperatures on Chinese rice wine quality were investigated. The compositions and concentrations of ethanol, sugars, glycerol, and organic acids in the mash of Chinese rice wine samples were determined by HPLC method. The highest ethanol concentration and the highest glycerol concentration both were attained at the fermentation mash treated at 23°C. The highest peak value of maltose (90 g/L was obtained at 18°C. Lactic acid and acetic acid both achieved maximum values at 33°C. The experimental results indicated that temperature contributed significantly to the ethanol production, acid flavor contents, and sugar contents in the fermentation broth of the Chinese rice wines.

  13. Study of dopant concentrations on thermal induced mode instability in high power fiber amplifiers

    CERN Document Server

    Tao, Rumao; Wang, Xiaolin; Zhou, Pu; Liu, Zejin

    2015-01-01

    Dependence of mode instabilities (MI) on ytterbium dopant concentrations in high power fiber amplifiers has been investigated. It is theoretically shown that, by only varying the fiber length to maintain the same total small-signal pump absorption, the MI threshold is independent of dopant concentration. MI thresholds of gain fibers with ytterbium dopant concentration of 5.93X10^25/m3 and 1.02X10^26/m3 have been measured, which exhibit similar thresholds and agree with theoretical results. The result indicates that heavy doping of active fiber can be adopted to suppress nonlinear effects without decreasing MI threshold, which provides a method of maximizing the power output of fiber laser, taking into account the stimulated Brillouin scattering, stimulated Raman Scattering, and MI thresholds simultaneously.

  14. Determination of Ge content in high concentration Ge-doped Czochralski Si single crystals by FTIR

    Institute of Scientific and Technical Information of China (English)

    JIANG Zhongwei; ZHANG Weilian; NIU Xinhuan

    2005-01-01

    SiGe single crystals with different Ge concentrations were measured by Fourier transform infrared (FTIR) spectroscopy at room temperature (RT) and 10 K. A new peak appears at the wave number of 710 cm-1 and the spectroscopy becomes clearer with an increase in Ge content. The absorption strength and wave sharp of the 710 cm-1 peak are independent of temperature. The relation of the absorption coefficient amax, the band width of half maximum (BWHM) W1/2 of the 710 cm-1 peak, and the Ge concentration is determined with the Ge content obtained by SEM-EDX. The conversion factor is k = 1.211 at 10 K. Therefore, the Ge content in high concentration Ge doped CZ-Si single crystals can be determined by FTIR.

  15. Thermodynamically consistent description of the hydrodynamics of free surfaces covered by insoluble surfactants of high concentration

    CERN Document Server

    Thiele, Uwe; Plapp, Mathis

    2012-01-01

    In this paper we propose several models that describe the dynamics of liquid films which are covered by a high concentration layer of insoluble surfactant. First, we briefly review the 'classical' hydrodynamic form of the coupled evolution equations for the film height and surfactant concentration that are well established for small concentrations. Then we re-formulate the basic model as a gradient dynamics based on an underlying free energy functional that accounts for wettability and capillarity. Based on this re-formulation in the framework of nonequilibrium thermodynamics, we propose extensions of the basic hydrodynamic model that account for (i) nonlinear equations of state, (ii) surfactant-dependent wettability, (iii) surfactant phase transitions, and (iv) substrate-mediated condensation. In passing, we discuss important differences to most of the models found in the literature.

  16. Biodegradation of high concentrations of phenol by baker’s yeast in anaerobic sequencing batch reactor

    Directory of Open Access Journals (Sweden)

    Ali Asghar Najafpoor

    2015-06-01

    Full Text Available Background: Phenol, as a pure substance, is used in many fields because of its disinfectant, germicidal, local anesthetic, and peptizing properties. Aqueous solutions of phenol are produced as waste in industries and discharged into the environment. Therefore, elevated concentrations of phenol may be found in air or water because of industrial discharge or the use of phenolic products. Method: The strains of Saccharomyces cerevisiae used in this project were natural strains previously purchased from Razavy company. They were grown at 30°C on Petri plates containing yeast extract glucose (YGC and then purified by being spread onto new plates, and isolated colonies were obtained. These colonies provided the basis of selection. Prepared strains were applied in anaerobic sequencing batch reactors (ASBRs as first seed. The experiment conditions were optimized using response surface methodology (RSM. After the determined runs were performed using Design-Expert software, data were analyzed using mentioned software as well. Results: This study evaluated the capability of baker’s yeast to remove phenol in high concentrations. The tested strains showed excellent tolerance to phenol toxicity at concentrations up to 6100 mg/L. Study of the batch degradation process showed that the phenol removal rate could exceed 99.9% in 24 hours at a concentration of 1000 mg/L. The results showed catechol is the first intermediate product of phenol degradation. In survey results of the Design–Expert software, R2 and Adeq precision were 0.97 and 25.65, respectively. Conclusion: The results demonstrated that ASBR performs robustly under variable influent concentrations of inhibitory compounds. The high removal performance despite the high phenol concentration may be a result of reactor operating strategies. Based on the progressive increase of inlet phenol concentration, allowing for an enhanced biomass acclimation in a short time, results at the microbiological levels

  17. Generation of high-titer viral preparations by concentration using successive rounds of ultracentrifugation

    Directory of Open Access Journals (Sweden)

    Ichim Christine V

    2011-08-01

    Full Text Available Abstract Background Viral vectors provide a method of stably introducing exogenous DNA into cells that are not easily transfectable allowing for the ectopic expression or silencing of genes for therapeutic or experimental purposes. However, some cell types, in particular bone marrow cells, dendritic cells and neurons are difficult to transduce with viral vectors. Successful transduction of such cells requires preparation of highly concentrated viral stocks, which permit a high virus concentration and multiplicity of infection (MOI during transduction. Pseudotyping with the vesicular stomatitis virus G (VSV-G envelope protein is common practice for both lentiviral and retroviral vectors. The VSV-G glycoprotein adds physical stability to retroviral particles, allowing concentration of virus by high-speed ultracentrifugation. Here we describe a method report for concentration of virus from large volumes of culture supernatant by means of successive rounds of ultracentrifugation into the same ultracentrifuge tube. Method Stable retrovirus producer cell lines were generated and large volumes of virus-containing supernatant were produced. We then tested the transduction ability of virus following varying rounds of concentration by ultra-centrifugation. In a second series of experiments lentivirus-containing supernatant was produced by transient transfection of 297T/17 cells and again we tested the transduction ability of virus following multiple rounds of ultra-centrifugation. Results We report being able to centrifuge VSV-G coated retrovirus for as many as four rounds of ultracentrifugation while observing an additive increase in viral titer. Even after four rounds of ultracentrifugation we did not reach a plateau in viral titer relative to viral supernatant concentrated to indicate that we had reached the maximum tolerated centrifugation time, implying that it may be possible to centrifuge VSV-G coated retrovirus even further should it be necessary

  18. ATP as an indicator of biomass activity in thermophilic upflow anaerobic sludge blanket reactor

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    This work investigated the biomass activity in a thermophilic upflow anaerobic sludge blanket (UASB) reactor of wastewater treatment. Synthetic textile wastewater with pH 10-11, COD level of 2000-3000 mg/L was tested. Cellular adenosine triphosphate (ATP) in volatile solids (VS; mg ATP/gVS) was measured and expressed as specific ATP content to compare the biomass activity in up zone and lower zone in UASB reactor. The result shows that the specific ATP content based on total volatile solids (VS)in lower zone (0. 046 mgATP/gVS average) is much lower than that in up zone (0.62 mgATP/gVS average) due to high content of inactive biomass and high pH in lower zone. The SATP in up zone increases as HRT increases and approaches to a maximum value of 0.85 mgATP/gVS at HRT of 7h, then decreases. It shows most of the total VS in up zone represent active bacterial biomass at HRT of 7h. Rate of subtract utilization is directly related to the activity of microorganisms in the reactor. The effect of HRT on SATP in lower zone is not as significant as on SATP in up zone. The buffer capacity of the thermophilic UASB reactor is very good. It is the activity of sludge granules in lower zone that give the UASB reactor such a good buffer capacity to the inlet high pH.

  19. Rapid and direct detection of attomole adenosine triphosphate (ATP) by MALDI-MS using rutile titania chips.

    Science.gov (United States)

    Manikandan, Muthu; Hasan, Nazim; Wu, Hui-Fen

    2012-11-07

    We report the rutile titania-based capture of ATP and its application as a MALDI-MS target plate. This chip, when immersed in solutions containing different concentrations of ATP, can capture ATP and lead to its successful detection in MALDI-MS. We have optimized the ideal surface, showing an increased capture efficacy of the 900 °C (rutile) titania surfaces. We demonstrate the use of this chip as a target plate for direct analysis of the attached ATP using MALDI-MS, down to attomolar concentrations. This chip has a promising future for the detection of ATP in environmental samples, which may eventually be used as a pollution indicator in particular environments.

  20. The effects of high temperature level on square Bt protein concentration of Bt cotton

    Institute of Scientific and Technical Information of China (English)

    WANG Jun; CHEN Yuan; YAO Meng-hao; LI Yuan; WEN Yu-jin; ZHANG Xiang; CHEN De-hua

    2015-01-01

    Higher bol worm survival rates were detected after high temperature presented during square period in Bt cotton. The objective of this study was to investigate the effects of high temperature level on the Bt efifcacy of two different types of Bt cotton cultivars at squaring stage. During the 2011 to 2013 cotton growth seasons, high temperature treatments ranged from 34 to 44°C in climate chambers, and ifeld experiments under high temperature weather with various temperature levels were conducted to investigate the effects of the high temperature level on square Bt protein concentration and nitrogen metabolism. The climate chamber experiments showed that the square insecticidal protein contents reduced after 24 h elevated temperature treatments for both cultivars, whereas signiifcant declines of the square insecticidal protein contents were detected at temperature >38°C, and only slightly numerical reductions were observed when temperature below 38°C. Similar high temperature responses were also observed at the two ifeld experimental sites in 2013. Correspondingly, high temperature below 38°C seems have little effect on the square amino acid concentrations, soluble protein contents, glutam-ic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminase (GOT) activities as wel as protease and peptidase activities; however, when the temperature was above 38°C, reduced soluble protein contents, enhanced amino acid con-centrations, decreased GPT and GOT activities, bolstered protease and peptidase activities in square were detected. In general, the higher the temperature is (>38°C), the larger the changes for the above compound contents and key enzymes activities of the square protein cycle. The ifndings indicated that the unstable insect resistance of the square was related to high temperature level during square stage.

  1. Gulf of Bothnia receives high concentrations of potentially toxic metals from acid sulphate soils

    Energy Technology Data Exchange (ETDEWEB)

    Roos, M.; Aastroem, M. [Kalmar Univ., Kalmar (Sweden)

    2006-07-01

    An estimated 460000 ha of acid sulphate soil (AS soil) occur within the river catchments bordering the Gulf of Bothnia in Finland and Sweden. This soil type exists because extensive areas of sulphide-bearing Holocene sediments have been drained for agricultural purposes, resulting in oxidation of metal sulphides to sulphuric acid and the concomitant formation of these acidic, environmentally-unfriendly soils. The aim of this study was to compare median values of metal concentrations in rivers discharging into the Gulf of Bothnia and obtain a uniform picture of to what extent these rivers are affected by AS-soil leaching. Dissolved element concentrations for arsenic (As), cadmium (Cd), chromium (Cr), iron (Fe), nickel (Ni), lead (Pb), and zinc (Zn) were determined in 47 rivers (catchment size > 500 km2) discharging into Gulf of Bothnia (a few into Gulf of Finland) along the coastline of Finland and Sweden. Water chemistry data was obtained from the Environmental Information System (HERTTA) database at the Finnish Environment Institute, the publicly available online database at Swedish University of Agricultural Sciences, Department of Environmental Assessment, and from a previous study of the authors. One area in central-western Finland proved to have highly elevated concentrations of Cd, Ni and Zn, and they all occurred with a similar spatial pattern and had the highest concentrations in rivers Teuvanjoki and Maalahdenjoki. This is caused by AS-soil leaching. The metalloid As and the metals Cr, Fe, Pb did not display this pattern and are, therefore, in line with previous studies, not leached abundantly from the AS soils, although they overall occur at somewhat higher concentrations in the Finnish as compared with those in the Swedish rivers. Thorough planning of land-use operations (e.g. land reclamation through ditching, dredging of rivers and estuaries, etc.) in AS-soil landscapes should be necessary, which is currently not the case, to reduce the high

  2. Impact of High Concentration Solutions on Hydraulic Properties of Geosynthetic Clay Liner Materials

    Directory of Open Access Journals (Sweden)

    Lei Liu

    2012-11-01

    Full Text Available This study focuses on the impact of landfill high concentration solutions erosion on geosynthetic clay liner (GCL materials permeability. The permeation tests on the GCL, submerged using different kinds of solutions with different concentrations, were carried out systematically by taking these chemical solutions as permeant liquids. Based on seasonal variations of ion concentrations in Chenjiachong landfill leachate (Wuhan Province, CaCl2, MgCl2, NaCl, and KCl were selected as chemical attack solutions to carry out experimental investigations under three concentrations (50 mM, 100 mM, 200 mM and soak times (5, 10, and 20 days. The variation law of the GCL hydraulic conductivity under different operating conditions was analyzed. The relationship between GCL hydraulic conductivity, chemical solutions categories, concentrations, and soak times were further discussed. The GCL hydraulic conductivity, when soaked and permeated with high concentration chemical solutions, increases several times or exceeds two orders of magnitude, as compared with the permeation test under normal conditions that used water as the permeant liquid. This reveals that GCL is very susceptible to chemical attack. For four chemical solutions, the chemical attack effect on GCL hydraulic conductivity is CaCl2 > MgCl2 > KCl > NaCl. The impact of soak times on GCL hydraulic conductivity is the cooperative contribution of the liner chemical attack reaction and hydration swelling. A longer soak time results in a more advantageous hydration swelling effect. The chemical attack reaction restrains the hydration swelling of the GCL. Moreover, the GCL hydraulic conductivity exponentially decreases with the increased amplitude of thickness.

  3. Calculation of multicomponent ionic diffusion from zero to high concentration: II. Inclusion of associated ion species

    Science.gov (United States)

    Felmy, Andrew R.; Weare, John H.

    1991-01-01

    This paper presents a theoretical model of multicomponent ionic diffusion which is valid to high concentration for systems which show ion association. The development is completely general for species which do not react with the solvent. It is demonstrated that the Onsager phenomenological coefficients for associated ion species can be unambiguously determined only in solutions where the concentration of these species can be independently measured. In all other instances, only combinations of the Onsager coefficients for the bare ions and the ion pairs can be experimentally determined. The results of our formulations are contrasted with those of more simplified models for systems containing Na 2SO 4 and MgSO 4, as well as for multicomponent natural seawater. The differences between our model and simplified models are significant, especially at high concentration. Inconsistencies which may develop with the use of the simplified approaches are demonstrated. Our approach requires considerable data which are not available at temperatures other than 25°C. Therefore, other approaches which are based only on data at infinite dilution are of great interest. We show here that, if chemical potential derivatives are included in the infinite dilution model of Nernst and Hartley which uses only infinite dilution mobilities, the model can be extended to slightly concentrated solutions. This extended Nernst-Hartley model gives good agreement with all of the existing experimental mutual diffusion coefficient data at concentrations below about 0.2 M in the six component system Na-K-Ca-Mg-Cl-SO 4-H 2O. This may be the most reliable way to extend infinite dilution data into more concentrated regions. In the systems we have studied, the inclusion of ion-association species for weakly interacting species does not appear to provide significant improvement over our generalized Nernst-Hartley model.

  4. Antibacterial properties of cyclodextrin-antiseptics-complexes determined by microplate laser nephelometry and ATP bioluminescence assay.

    Science.gov (United States)

    Finger, Susanne; Wiegand, Cornelia; Buschmann, Hans-Jürgen; Hipler, Uta-Christina

    2013-08-16

    Cyclodextrins (CDs) are able to form inclusion complexes with other molecules, thereby, protecting these guest molecules from degradation, enhancing their biocompatibility or influencing their physiological distribution while retaining their activity. Here, antibacterial effects of CD-complexes with the antiseptics chlorhexidine diacetate (CHX), iodine (IOD) and polihexanide (PHMB) were determined using two different in vitro methods, microplate laser nephelometry and an ATP bioluminescence assay. Laser nephelometry is a direct method for monitoring and evaluating growth of micro-organisms by measurement of the turbidity of the solution. In contrast, the ATP bioluminescence assay determines specifically the amount of metabolic active bacterial cells. The antibacterial effects of CD-antiseptics-complexes were examined for Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus and Staphylococcus epidermidis and the results of both methods were compared in respect of calculated means of half maximal inhibitory concentrations (IC50) and statistical evaluated Pearson's correlation coefficients (r). It could be demonstrated that both methods showed a high comparability although they differ in the parameters tested. This study revealed that CD-complexes with CHX and PHMB were most effective against E. coli and the tested staphylococci. While CD-IOD-complexes obtained high activity against K. pneumoniae, P. aeruginosa was distinctly more resistant compared to the other bacteria.

  5. Metal Induced Inhibition of Photosynthesis,Photosynthetic Electron Transport Chain and ATP Content of Anabaena doliolum and Chlorella vulgaris:Interaction with Exogenous ATP

    Institute of Scientific and Technical Information of China (English)

    NIRUPAMAMALLICK; L.C.RAI

    1992-01-01

    This study demonstrates a concentration dependent inhibition of carbon fixation,O2 evolution,photosynthetic electron transport chain and ATP content of A.doliolum and C.vulgaris by Cu,Ni and Fe.Although the mode of inhibition of photosynthetic electron transport chain of both the algae was similar.PS Ⅱdepicted greater sensitivity to the test metals used.The toxicity in both organisms was Cu>Ni>Fe.A.doliolum was,however,more sensitive to Cu and Ni,and C.vulgaris to Fe.Toxicity was generally dependent on metal uptake,which in turn was dependent on their concentrations in the external medium.A partial restoration of nutrient uptake,carbon fixation,and enzyme activities following supplementation of exogenous ATP suggests that ATP regulates toxicity through chelation.

  6. Electrophysiology of autonomic neuromuscular transmission involving ATP.

    Science.gov (United States)

    Sneddon, P

    2000-07-01

    Electrophysiological investigations of autonomic neuromuscular transmission have provided great insights into the role of ATP as a neurotransmitter. Burnstock and Holman made the first recordings of excitatory junction potentials (e.j.p.s) produced by sympathetic nerves innervating the smooth muscle of the guinea-pig vas deferens. This led to the identification of ATP as the mediator of e.j.p.s in this tissue, where ATP acts as a cotransmitter with noradrenaline. The e.j.p.s are mediated solely by ATP acting on P2X(1) receptors leading to action potentials and a rapid phasic contraction, whilst noradrenaline mediates a slower, tonic contraction which is not dependent on membrane depolarisation. Subsequent electrophysiological studies of the autonomic innervation of smooth muscles of the urogenital, gastrointestinal and cardiovascular systems have revealed a similar pattern of response, where ATP mediates a fast electrical and mechanical response, whilst another transmitter such as noradrenaline, acetylcholine, nitric oxide or a peptide mediates a slower response. The modulation of junction potentials by a variety of pre-junctional receptors and the mechanism of inactivation of ATP as a neurotransmitter will also be described.

  7. Continuous and real-time bioaerosol monitoring by combined aerosol-to-hydrosol sampling and ATP bioluminescence assay.

    Science.gov (United States)

    Park, Ji-Woon; Kim, Hyeong Rae; Hwang, Jungho

    2016-10-19

    We present a methodology for continuous and real-time bioaerosol monitoring wherein an aerosol-to-hydrosol sampler is integrated with a bioluminescence detector. Laboratory test was conducted by supplying an air flow with entrained test bacteria (Staphylococcus epidermidis) to the inlet of the sampler. High voltage was applied between the discharge electrode and the ground electrode of the sampler to generate air ions by corona discharge. The bacterial aerosols were charged by the air ions and sampled in a flowing liquid containing both a cell lysis buffer and adenosine triphosphate (ATP) bioluminescence reagents. While the liquid was delivered to the bioluminescence detector, sampled bacteria were dissolved by the cell lysis buffer and ATP was extracted. The ATP was reacted with the ATP bioluminescence reagents, causing light to be emitted. When the concentration of bacteria in the aerosols was varied, the ATP bioluminescence signal in relative light units (RLUs) closely tracked the concentration in particles per unit air volume (# cm(-3)), as measured by an aerosol particle sizer. The total response time required for aerosol sampling and ATP bioluminescence detection increased from 30 s to 2 min for decreasing liquid sampling flow rate from 800 to 200 μLPM, respectively. However, lower concentration of S. epidermidis aerosols was able to be detected with lower liquid sampling flow rate (1 RLU corresponded to 6.5 # cm(-3) of S. epidermidis aerosols at 200 μLPM and 25.5 # cm(-3) at 800 μLPM). After obtaining all data sets of concentration of S. epidermidis aerosols and concentration of S. epidermidis particles collected in the flowing liquid, it was found that with our bioluminescence detector, 1 RLU corresponded to 1.8 × 10(5) (±0.2 × 10(5)) # mL(-1) of S. epidermidis in liquid. After the lab-test with S. epidermidis, our bioaerosol monitoring device was located in the lobby of a building. Air sampling was conducted continuously for 90

  8. Thermal stability of high concentration lysozyme across varying pH: A Fourier Transform Infrared study

    Directory of Open Access Journals (Sweden)

    Sathyadevi Venkataramani

    2013-01-01

    Full Text Available Aim: The current work is aimed at understanding the effect of pH on the thermal stability of hen egg white lysozyme (HEWL at high concentration (200 mg/mL. Materials and Methods: Fourier Transform Infrared (FTIR Spectroscopy with modified hardware and software to overcome some of the traditional challenges like water subtraction, sample evaporation, proper purging etc., are used in this study. Results: HEWL was subjected to thermal stress at pH 3.0-7.0 between 25°C and 95°C and monitored by FTIR spectroscopy. Calculated T m values showed that the enzyme exhibited maximum thermal stability at pH 5.0. Second derivative plots constructed in the amide I region suggested that at pH 5.0 the enzyme possessed higher amount of α-helix and lower amount of aggregates, when compared to other pHs. Conclusions: Considering the fact that HEWL has attractive applications in various industries and being processed under different experimental conditions including high temperatures, our work is able to reveal the reason behind the pH dependent thermal stability of HEWL at high concentration, when subjected to heat denaturation. In future, studies should aim at using various excipients that may help to increase the stability and activity of the enzyme at this high concentration.

  9. Carbon assimilation in Eucalyptus urophylla grown under high atmospheric CO2 concentrations: A proteomics perspective.

    Science.gov (United States)

    Santos, Bruna Marques Dos; Balbuena, Tiago Santana

    2017-01-06

    Photosynthetic organisms may be drastically affected by the future climate projections of a considerable increase in CO2 concentrations. Growth under a high concentration of CO2 could stimulate carbon assimilation-especially in C3-type plants. We used a proteomics approach to test the hypothesis of an increase in the abundance of the enzymes involved in carbon assimilation in Eucalyptus urophylla plants grown under conditions of high atmospheric CO2. Our strategy allowed the profiling of all Calvin-Benson cycle enzymes and associated protein species. Among the 816 isolated proteins, those involved in carbon fixation were found to be the most abundant ones. An increase in the abundance of six key enzymes out of the eleven core enzymes involved in carbon fixation was detected in plants grown at a high CO2 concentration. Proteome changes were corroborated by the detection of a decrease in the stomatal aperture and in the vascular bundle area in Eucalyptus urophylla plantlets grown in an environment of high atmospheric CO2. Our proteomics approach indicates a positive metabolic response regarding carbon fixation in a CO2-enriched atmosphere. The slight but significant increase in the abundance of the Calvin enzymes suggests that stomatal closure did not prevent an increase in the carbon assimilation rates.

  10. Lower Salinomycin Concentration Increases Apoptotic Detachment in High-Density Cancer Cells

    Directory of Open Access Journals (Sweden)

    Sungpil Yoon

    2012-10-01

    Full Text Available The present study identified a novel salinomycin (Sal sensitization mechanism in cancer. We tested whether Sal reduced proliferation in a high-density population by counting attached cell numbers after Sal treatment. Sal reduced proliferation in high-density cell populations. Longer exposure to Sal further reduced proliferation. Sal concentrations of 0.1 and 5 μM had similar sensitization effects, suggesting that Sal toxicity was minimal with longer exposure to a high-density cell population. The results suggest that Sal can be applied at a relatively low concentration for a longer time to overcome drug-resistant solid tumors. The 0.5 μM Sal treatment resulted in fewer attached cells than that of the 5 μM Sal treatment with a longer exposure. The lower Sal concentration mainly increased the number of easily detachable cells on the surface. In particular, 0.5 μM Sal increased cellular detachment of newly produced daughter cells. The easily-detachable cells were undergoing apoptosis. It seems that the 0.5 μM Sal treatment also increased cellular toxicity. These novel findings may contribute to the development of Sal-based therapy for patients with drug-resistant cancer or a high-density solid tumor.

  11. Anaerobic biotransformation of high concentrations of chloroform by an enrichment culture and two bacterial isolates.

    Science.gov (United States)

    Shan, Huifeng; Kurtz, Harry D; Mykytczuk, Nadia; Trevors, Jack T; Freedman, David L

    2010-10-01

    A fermentative enrichment culture (designated DHM-1) was developed that is capable of cometabolically biotransforming high concentrations of chloroform (CF) to nontoxic end products. Two Pantoea spp. were isolated from DHM-1 that also possess this dechlorination capability. Following acclimation to increasing levels of CF, corn syrup-grown DHM-1 was able to transform over 500 mg/liter CF in the presence of vitamin B(12) (approximately 3% of CF on a molar basis) at a rate as high as 22 mg/liter/day in a mineral salts medium. CO, CO(2), and organic acids were the predominant biodegradation products, suggesting that hydrolytic reactions predominate during CF transformation. DHM-1 was capable of growing on corn syrup in the presence of high concentrations of CF (as may be present near contaminant source zones in groundwater), which makes it a promising culture for bioaugmentation. Strains DHM-1B and DHM-1T transform CF at rates similar to that of the DHM-1 enrichment culture. The ability of these strains to grow in the presence of high concentrations of CF appears to be related to alteration of membrane fluidity or homeoviscous and homeophasic adaptation.

  12. Effects of High Ammonium Concentration on Growth and Nutrient Uptake of Lettuce Plants with Solution Culture

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    A nutrition solution experiment was conducted over two months to investigate the response of vegetable crops to high concentrations of ammonium, using lettuce (Lactuca sativa L. cv. Angustana Irish) as a test crop. Ammonium concentrations were designed in 5 levels, ranging from 12 mmol N L-1 to 22 mmol N L-1 and local tap water was used as water source. At the first culture stage (0-9 days), lettuce plants maintained normal growth while the lettuce roots were increasingly impaired. During the subsequent three stages the root structure was greatly damaged, and roots became brown or black through continuous supply of high concentration of ammonium. However, there was no obvious reduction of the aboveground biomass of the plants in the high ammonium treatments compared to those supplied with nitrate alone. In contrast to results obtained in another experiment from us with distilled water, the detrimental effect of high ammonium concentration on lettuce growth was greatly alleviated. Based on the results, it was postulated that the small amount of nitrate and the higher amount of bicarbonate existed in the tap water might mitigate the adverse effects of high ammonium N. The higher bicarbonate content in water and soil has usually been regarded as a major constraint factor limiting plant growth in calcareous soil areas. However, the reaction of bicarbonate to ammonium might produce positively interactive effect on reduction of both damages. The lettuce plants grown in ammonium solutions took up less P, K, Fe, Mn and Cu and more Ca than those grown in the nitrate nutrient solution. In conclusion, the results indicated that the N form imposed an obvious influence on absorption of cations and anions. Supplying ammonium-N stimulated transport of Ca, Mg and Mn to shoots of lettuce.

  13. High variability of dissolved iron concentrations in the vicinity of Kerguelen Island (Southern Ocean

    Directory of Open Access Journals (Sweden)

    F. Quéroué

    2015-01-01

    Full Text Available Dissolved Fe (dFe concentrations were measured in the upper 1300 m of the water column in the vicinity of Kerguelen Island as part of the second Kerguelen Ocean Plateau compared Study (KEOPS2. Concentrations ranged from 0.06 nmol L−1 in offshore, Southern Ocean waters, to 3.82 nmol L−1 within Hillsborough Bay, on the north-eastern coast of Kerguelen Island. Direct island runoff, glacial melting and resuspended sediments were identified as important inputs of dFe that could potentially fertilize the northern part of the plateau. A significant deep dFe enrichment was observed over the plateau with dFe concentrations increasing up to 1.30 nmol L−1 close to the seafloor, probably due to sediment resuspension and pore water release. Biological uptake was identified as a likely explanation for the decrease in dFe concentrations between two visits (28 days apart at a station above the plateau. Our results allowed studying other processes and sources, such as atmospheric inputs, lateral advection of enriched seawater, remineralization processes and the influence of the Polar Front (PF as a vector for Fe transport. Overall, heterogeneous sources of Fe over and off the Kerguelen Plateau, in addition to strong variability in Fe supply by vertical or horizontal transport, may explain the high variability in dFe concentrations observed during this study.

  14. Cardiac specific ATP-sensitive K+ channel (KATP) overexpression results in embryonic lethality.

    Science.gov (United States)

    Toib, Amir; Zhang, Hai Xia; Broekelmann, Thomas J; Hyrc, Krzysztof L; Guo, Qiusha; Chen, Feng; Remedi, Maria S; Nichols, Colin G

    2012-09-01

    Transgenic mice overexpressing SUR1 and gain of function Kir6.2[∆N30, K185Q] K(ATP) channel subunits, under cardiac α-myosin heavy chain (αMHC) promoter control, demonstrate arrhythmia susceptibility and premature death. Pregnant mice, crossed to carry double transgenic progeny, which harbor high levels of both overexpressed subunits, exhibit the most extreme phenotype and do not deliver any double transgenic pups. To explore the fetal lethality and embryonic phenotype that result from K(ATP) overexpression, wild type (WT) and K(ATP) overexpressing embryonic cardiomyocytes were isolated, cultured and voltage-clamped using whole cell and excised patch clamp techniques. Whole mount embryonic imaging, Hematoxylin and Eosin (H&E) and α smooth muscle actin (αSMA) immunostaining were used to assess anatomy, histology and cardiac development in K(ATP) overexpressing and WT embryos. Double transgenic embryos developed in utero heart failure and 100% embryonic lethality by 11.5 days post conception (dpc). K(ATP) currents were detectable in both WT and K(ATP)-overexpressing embryonic cardiomyocytes, starting at early stages of cardiac development (9.5 dpc). In contrast to adult cardiomyocytes, WT and K(ATP)-overexpressing embryonic cardiomyocytes exhibit basal and spontaneous K(ATP) current, implying that these channels may be open and active under physiological conditions. At 9.5 dpc, live double transgenic embryos demonstrated normal looping pattern, although all cardiac structures were collapsed, probably representing failed, non-contractile chambers. In conclusion, K(ATP) channels are present and active in embryonic myocytes, and overexpression causes in utero heart failure and results in embryonic lethality. These results suggest that the K(ATP) channel may have an important physiological role during early cardiac development.

  15. The P2X7 receptor is an important regulator of extracellular ATP levels

    Directory of Open Access Journals (Sweden)

    Andrea eBrandao-Burch

    2012-03-01

    Full Text Available Controlled ATP release has been demonstrated from many neuronal and non-neuronal cell types. Once released, extracellular ATP acts on cells in a paracrine manner via purinergic receptors. Considerable evidence now suggests that extracellular nucleotides, signalling via P2 receptors, play important roles in bone homeostasis modulating both osteoblast and osteoclast function. In this study, we demonstrate that mouse osteoclasts and their precursors constitutively release ATP into their extracellular environment. Levels were highest at day 2 (precursor cells, possibly reflecting the high number of red blood cells and accessory cells present. Mature osteoclasts constitutively released ATP in the range 0.05-0.5pmol/ml/cell. Both osteoclasts and osteoblasts express mRNA and protein for the P2X7 receptor. We found that in osteoclasts, expression levels are 4-fold higher in mature cells relative to precursors, whilst in osteoblasts expression remains relatively constant during differentiation. Selective antagonists (0.1-100µM AZ10606120, A438079 and KN-62 were used to determine whether this release was mediated via P2X7 receptors. AZ10606120, A438079 and KN-62, at 0.1-10µM, decreased ATP release by mature osteoclasts by up to 70%, 60% and 80%, respectively. No differences in cell viability were observed. ATP release also occurs via vesicular exocytosis; inhibitors of this process (1-100µM NEM or brefeldin A had no effect on ATP release from osteoclasts. P2X7 receptor antagonists (0.1-10µM also decreased ATP release from primary rat osteoblasts by up to 80%. These data show that ATP release via the P2X7 receptor contributes to extracellular ATP levels in osteoclast and osteoblast cultures, suggesting an important additional role for this receptor in autocrine/paracrine purinergic signalling in bone.

  16. Involvement of connexin43 hemichannel in ATP release after γ-irradiation

    Science.gov (United States)

    Ohshima, Yasuhiro; Tsukimoto, Mitsutoshi; Harada, Hitoshi; Kojima, Shuji

    2012-01-01

    Ionizing radiation induces biological effects not only in irradiated cells but also in non-irradiated cells, which is called the bystander effect. Recently, in vivo and in vitro experiments have suggested that both gap junction hemichannel connexin43 (Cx43) and extracellular adenosine triphosphate (ATP) released from cells play a role in the bystander effect. We have reported that γ-irradiation induces ATP release from B16 melanoma cells, which is dependent on the P2X7 receptor. However, the mechanism of ATP release caused by irradiation remains unclear. We here show the involvement of Cx43 in P2X7 receptor-dependent ATP release after 0.5 Gy γ-irradiation. Inhibitors of gap junction hemichannels and an inhibitory peptide for Cx43 (gap26), but not an inhibitory peptide for pannexin1 (Panx1), significantly blocked γ-irradiation-induced ATP release from B16 melanoma cells. We confirmed high expression of Cx43 mRNA in B16 melanoma cells. These results suggest involvement of Cx43 in radiation-induced ATP release. We found that after 0.5 Gy γ-irradiation tyrosine phosphorylation was significantly blocked by P2X7 receptor antagonist, but not gap26, suggesting that tyrosine phosphorylation is a downstream event from the P2X7 receptor. Since tyrosine kinase inhibitor significantly suppressed radiation-induced ATP release, tyrosine phosphorylation appears to play an important role in the Cx43-mediated ATP release downstream of the P2X7 receptor. In conclusion, the Cx43 hemichannel, which lies downstream of the P2X7 receptor, is involved in ATP release in response to radiation. Our results suggest a novel mechanism for radiation-induced biological effects mediated by both ATP and Cx43. PMID:22843620

  17. Deer exposed to exceptionally high concentrations of lead near the continental mine in Idaho, USA

    Science.gov (United States)

    Beyer, W.N.; Gaston, G.; Brazzle, R.; O'Connell, A.F.; Audet, D.J.

    2007-01-01

    Habitat surrounding the inactive Continental Mine in northern Idaho, USA, supports bear (Ursus arctos, Ursus americanus), moose (Alces alces), elk (Cervus elaphus), woodland caribou (Rangifer tarandus caribou), and abundant mule (Odocoileus hemionus) and white-tailed deer (Odocoileus virginianus). Tailings on the mining site were capped and remediated in 2003 to reduce environmental exposure of surrounding soil and sediments of Blue Joe Creek, downslope of the mine. Before capping, the mean Pb concentration in deer pellets collected on-site was 920 mg/kg of Pb (dry wt). This exposure, if chronic, would be comparable to an exposure that could be lethal to cattle or horses. Surprisingly, the mean pellet Pb concentration of 950 mg/kg in 2004 was as high as it was before remediation, and it was related to a high rate of soil ingestion. Mean soil content of the pellets collected from the capped site in 2004 was 22% dry weight, estimated from the acid-insoluble ash, a marker of soil ingestion. Clumps of sand and bits of rock were observed inside some of the pellets, and Pb concentrations in the pellets were correlated (p soil content. Although terrestrial risk assessments generally estimate exposure from diets and from incidentally ingested soil, the deer at this site were directly ingesting contaminated soil or mining waste. The mean Pb concentration of this ingested soil was estimated as 6,700 mg/kg and the maximum as 25,000 mg/kg, well above the Pb concentrations measured in the remediated cap. The deer seemed to be ingesting soil or mining waste from one or more small but highly contaminated sources located beyond the remediated cap.

  18. Power Stroke Angular Velocity Profiles of Archaeal A-ATP Synthase Versus Thermophilic and Mesophilic F-ATP Synthase Molecular Motors.

    Science.gov (United States)

    Sielaff, Hendrik; Martin, James; Singh, Dhirendra; Biuković, Goran; Grüber, Gerhard; Frasch, Wayne D

    2016-12-02

    The angular velocities of ATPase-dependent power strokes as a function of the rotational position for the A-type molecular motor A3B3DF, from the Methanosarcina mazei Gö1 A-ATP synthase, and the thermophilic motor α3β3γ, from Geobacillus stearothermophilus (formerly known as Bacillus PS3) F-ATP synthase, are resolved at 5 μs resolution for the first time. Unexpectedly, the angular velocity profile of the A-type was closely similar in the angular positions of accelerations and decelerations to the profiles of the evolutionarily distant F-type motors of thermophilic and mesophilic origins, and they differ only in the magnitude of their velocities. M. mazei A3B3DF power strokes occurred in 120° steps at saturating ATP concentrations like the F-type motors. However, because ATP-binding dwells did not interrupt the 120° steps at limiting ATP, ATP binding to A3B3DF must occur during the catalytic dwell. Elevated concentrations of ADP did not increase dwells occurring 40° after the catalytic dwell. In F-type motors, elevated ADP induces dwells 40° after the catalytic dwell and slows the overall velocity. The similarities in these power stroke profiles are consistent with a common rotational mechanism for A-type and F-type rotary motors, in which the angular velocity is limited by the rotary position at which ATP binding occurs and by the drag imposed on the axle as it rotates within the ring of stator subunits.

  19. Enrichment and isolation of Flavobacterium strains with tolerance to high concentrations of cesium ion.

    Science.gov (United States)

    Kato, Souichiro; Goya, Eri; Tanaka, Michiko; Kitagawa, Wataru; Kikuchi, Yoshitomo; Asano, Kozo; Kamagata, Yoichi

    2016-02-17

    Interest in the interaction of microorganisms with cesium ions (Cs(+)) has arisen, especially in terms of their potent ability for radiocesium bioaccumulation and their important roles in biogeochemical cycling. Although high concentrations of Cs(+) display toxic effects on microorganisms, there have been only limited reports for Cs(+)-tolerant microorganisms. Here we report enrichment and isolation of Cs(+)-tolerant microorganisms from soil microbiota. Microbial community analysis revealed that bacteria within the phylum Bacteroidetes, especially Flavobacterium spp., dominated in enrichment cultures in the medium supplemented with 50 or 200 mM Cs(+), while Gammaproteobacteria was dominant in the control enrichment cultures (in the presence of 50 and 200 mM K(+) instead of Cs(+)). The dominant Flavobacterium sp. was successfully isolated from the enrichment culture and was closely related to Flavobacterium chungbukense with 99.5% identity. Growth experiments clearly demonstrated that the isolate has significantly higher tolerance to Cs(+) compared to its close relatives, suggesting the Cs(+)-tolerance is a specific trait of this strain, but not a universal trait in the genus Flavobacterium. Measurement of intracellular K(+) and Cs(+) concentrations of the Cs(+)-tolerant isolate and its close relatives suggested that the ability to maintain low intracellular Cs(+) concentration confers the tolerance against high concentrations of external Cs(+).

  20. High Pb concentration stress on Typha latifolia growth and Pb removal in microcosm wetlands.

    Science.gov (United States)

    Han, Jianqiu; Chen, Fengzhen; Zhou, Yumei; Wang, Chaohua

    2015-01-01

    When constructed wetlands are used to treat high-Pb wastewater, Pb may become a stress to wetland plants, which subsequently reduces treatment performance and the other ecosystem services. To facilitate the design and operation of constructed wetlands for treatment of Pb-rich wastewater, we investigated the irreversible inhibitory level of Pb for Typha latifolia through experiments in microcosm wetlands. Seven horizontal subsurface flow constructed wetlands were built with rectangular plastic tanks and packed with marble chips and sand. All wetlands were transplanted with nine stems of Typha latifolia each. The wetlands were batch operated in a greenhouse with artificial wastewater (10 L each) for 12 days. Influent to the seven wetlands had different concentrations of Pb: 0 mg/L, 10 mg/L, 25 mg/L, 50 mg/L, 100 mg/L, 200 mg/L, and 500 mg/L, respectively. The results suggested that leaf chlorophyll relative content, relative growth rate, photosynthetic characteristics, activities of superoxide dismutase, peroxidase, and content of malondialdehyde were not affected when initial Pb concentration was at 100 mg/L and below. But when initial Pb concentration was above 100 mg/L, all of them were seriously affected. We conclude that high Pb concentrations wastewater could inhibit the growth of Typha latifolia and decrease the removal rate of wetlands.