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Sample records for continuous flow-type hyperpolarizing

  1. Magnetic resonance imaging of dissolved hyperpolarized 129Xe using a membrane-based continuous flow system

    NASA Astrophysics Data System (ADS)

    Amor, N.; Zänker, P. P.; Blümler, P.; Meise, F. M.; Schreiber, L. M.; Scholz, A.; Schmiedeskamp, J.; Spiess, H. W.; Münnemann, K.

    2009-11-01

    A technique for continuous production of solutions containing hyperpolarized 129Xe is explored for MRI applications. The method is based on hollow fiber membranes which inhibit the formation of foams and bubbles. A systematic analysis of various carrier agents for hyperpolarized 129Xe has been carried out, which are applicable as contrast agents for in vivo MRI. The image quality of different hyperpolarized Xe solutions is compared and MRI results obtained in a clinical as well as in a nonclinical MRI setting are provided. Moreover, we demonstrate the application of 129Xe contrast agents produced with our dissolution method for lung MRI by imaging hyperpolarized 129Xe that has been both dissolved in and outgassed from a carrier liquid in a lung phantom, illustrating its potential for the measurement of lung perfusion and ventilation.

  2. [Continuous dry fermentation of pig manure using up plug-flow type anaerobic reactor].

    PubMed

    Chen, Chuang; Deng, Liang-Wei; Xin, Xin; Zheng, Dan; Liu, Yi; Kong, Chui-Xue

    2012-03-01

    To solve the problems of ammonia inhibition and discharging difficulty in continuous dry fermentation of pig manure, under the experimental conditions of temperature of (25 +/- 2) degrees C and organic loading rate (TS) of 4.44 g x (L x d) (-1), a lab-scale up plug-flow type anaerobic reactor (UPAR) was setup to investigate biogas production, ammonia inhibition, effluent liquidity, and the feasibility of continuous dry fermentation of pig manure using up plug-flow type anaerobic reactor. The experiment was operated for 160 days using the pig manure with four different TS mass fractions (20%, 25%, 30%, 35%) as feeding. Results showed that the feeding TS mass fraction exerted a significant influence on the dry fermentation of pig manure; the stable volumetric biogas production rates of four different feeding TS mass fractions were 2.40, 1.73, 0.89, and 0.62 L x (L x d)(-1), respectively; the biogas producing efficiencies of the reactors with feeding TS mass fractions of 20%, 25% and 30% were obviously superior to that with feeding TS of 35%. With feeding TS mass fraction increased from 20% to 35%, obvious inhibition to biogas producing occurred when concentration of ammonia nitrogen reached more than 2 300 mg x L(-1). When the feeding TS mass fraction was 35%, the concentration of ammonia nitrogen could accumulate to 3 800 mg x L(-1) but biogas production rate decreased 74.1% of that with feeding TS of 20%. Additionally, while the feeding TS mass fraction was 35%, the effluent TS mass fraction achieved 17.1%, and the velocity of effluent was less than 0.002 m x s(-1) the effluent of UPAR could not be smoothly discharged.

  3. Molecular MRI in the Earth's Magnetic Field Using Continuous Hyperpolarization of a Biomolecule in Water.

    PubMed

    Rovedo, Philipp; Knecht, Stephan; Bäumlisberger, Tim; Cremer, Anna Lena; Duckett, Simon B; Mewis, Ryan E; Green, Gary G R; Burns, Michael; Rayner, Peter J; Leibfritz, Dieter; Korvink, Jan G; Hennig, Jürgen; Pütz, Gerhard; von Elverfeldt, Dominik; Hövener, Jan-Bernd

    2016-06-30

    In this work, we illustrate a method to continuously hyperpolarize a biomolecule, nicotinamide, in water using parahydrogen and signal amplification by reversible exchange (SABRE). Building on the preparation procedure described recently by Truong et al. [ J. Phys. Chem. B , 2014 , 118 , 13882 - 13889 ], aqueous solutions of nicotinamide and an Ir-IMes catalyst were prepared for low-field NMR and MRI. The (1)H-polarization was continuously renewed and monitored by NMR experiments at 5.9 mT for more than 1000 s. The polarization achieved corresponds to that induced by a 46 T magnet (P = 1.6 × 10(-4)) or an enhancement of 10(4). The polarization persisted, although reduced, if cell culture medium (DPBS with Ca(2+) and Mg(2+)) or human cells (HL-60) were added, but was no longer observable after the addition of human blood. Using a portable MRI unit, fast (1)H-MRI was enabled by cycling the magnetic field between 5 mT and the Earth's field for hyperpolarization and imaging, respectively. A model describing the underlying spin physics was developed that revealed a polarization pattern depending on both contact time and magnetic field. Furthermore, the model predicts an opposite phase of the dihydrogen and substrate signal after one exchange, which is likely to result in the cancelation of some signal at low field.

  4. Continuously Infusing Hyperpolarized 129Xe into Flowing Aqueous Solutions Using Hydrophobic Gas Exchange Membranes

    PubMed Central

    Cleveland, Zackary I.; Möller, Harald E.; Hedlund, Laurence W.; Driehuys, Bastiaan

    2009-01-01

    Hyperpolarized (HP) 129Xe yields high signal intensities in magnetic resonance (MR) and, through its large chemical shift range of ∼300 ppm, provides detailed information about the local chemical environment. To exploit these properties in aqueous solutions and living tissues requires the development of methods for efficiently dissolving HP 129Xe over an extended time period. To this end, we have used commercially available gas exchange modules to continuously infuse concentrated HP 129Xe into flowing liquids, including rat whole blood, for periods as long as one hour, and have demonstrated the feasibility of dissolved-phase MR imaging with sub-millimeter resolution within minutes. These modules, which exchange gases using hydrophobic microporous polymer membranes, are compatible with a variety of liquids and are suitable for infusing HP 129Xe into the bloodstream in vivo. Additionally, we have developed a detailed mathematical model of the infused HP 129Xe signal dynamics that should be useful in designing improved infusion systems that yield even higher dissolved HP 129Xe signal intensities. PMID:19702286

  5. A continuous-flow, high-throughput, high-pressure parahydrogen converter for hyperpolarization in a clinical setting.

    PubMed

    Hövener, Jan-Bernd; Bär, Sébastien; Leupold, Jochen; Jenne, Klaus; Leibfritz, Dieter; Hennig, Jürgen; Duckett, Simon B; von Elverfeldt, Dominik

    2013-02-01

    Pure parahydrogen (pH(2) ) is the prerequisite for optimal pH(2) -based hyperpolarization experiments, promising approaches to access the hidden orders of magnitude of MR signals. pH(2) production on-site in medical research centers is vital for the proliferation of these technologies in the life sciences. However, previously suggested designs do not meet our requirements for safety or production performance (flow rate, pressure or enrichment). In this article, we present the safety concept, design and installation of a pH(2) converter, operated in a clinical setting. The apparatus produces a continuous flow of four standard liters per minute of ≈98% enriched pH(2) at a pressure maximum of 50 bar. The entire production cycle, including cleaning and cooling to 25 K, takes less than 5 h, only ≈45 min of which are required for actual pH(2) conversion. A fast and simple quantification procedure is described. The lifetimes of pH(2) in a glass vial and aluminum storage cylinder are measured to be T(1C) (glass vial) =822 ± 29 min and T(1C) (Al cylinder) =129 ± 36 days, thus providing sufficiently long storage intervals and allowing the application of pH(2) on demand. A dependence of line width on pH(2) enrichment is observed. As examples, (1) H hyperpolarization of pyridine and (13) C hyperpolarization of hydroxyethylpropionate are presented.

  6. Production of carbonaceous adsorbents from agricultural by-products and novolac resin under a continuous countercurrent flow type pyrolysis operation.

    PubMed

    Ioannou, Z; Simitzis, J

    2013-02-01

    Carbonaceous adsorbents based on novolac resin (N) and olive stone biomass (B) in a proportion of 20/80 and 40/60 w./w. N/O were produced. The specimens were cured (c) and pyrolyzed/carbonized (C) up to 1000 °C under a continuous countercurrent flow type pyrolysis operation (N20B-cC, N40B-cC). Commercial activated carbon (AC) was used for comparison reasons. Methylene blue adsorption from its aqueous solutions onto the adsorbents and kinetic analysis were investigated. The specific surface area of adsorbents and the gross calorific values (GCV) of cured materials were determined. The results show that N40B-cC presents lower weight loss and shrinkage but higher methylene blue adsorption than N20B-cC. Pseudo-second order mechanism describes better methylene blue adsorption onto all adsorbents. The specific surface area of carbonaceous and the gross calorific values of cured materials follow the order: AC>N20B-cC>N40B-cC and N100-c>N40B-c>N20B-c>B respectively. Olive stone biomass may constitute a suitable precursor for the production of carbonaceous materials.

  7. Transportable hyperpolarized metabolites

    NASA Astrophysics Data System (ADS)

    Ji, Xiao; Bornet, Aurélien; Vuichoud, Basile; Milani, Jonas; Gajan, David; Rossini, Aaron J.; Emsley, Lyndon; Bodenhausen, Geoffrey; Jannin, Sami

    2017-01-01

    Nuclear spin hyperpolarization of 13C-labelled metabolites by dissolution dynamic nuclear polarization can enhance the NMR signals of metabolites by several orders of magnitude, which has enabled in vivo metabolic imaging by MRI. However, because of the short lifetime of the hyperpolarized magnetization (typically <1 min), the polarization process must be carried out close to the point of use. Here we introduce a concept that markedly extends hyperpolarization lifetimes and enables the transportation of hyperpolarized metabolites. The hyperpolarized sample can thus be removed from the polarizer and stored or transported for use at remote MRI or NMR sites. We show that hyperpolarization in alanine and glycine survives 16 h storage and transport, maintaining overall polarization enhancements of up to three orders of magnitude.

  8. Transportable hyperpolarized metabolites

    PubMed Central

    Ji, Xiao; Bornet, Aurélien; Vuichoud, Basile; Milani, Jonas; Gajan, David; Rossini, Aaron J.; Emsley, Lyndon; Bodenhausen, Geoffrey; Jannin, Sami

    2017-01-01

    Nuclear spin hyperpolarization of 13C-labelled metabolites by dissolution dynamic nuclear polarization can enhance the NMR signals of metabolites by several orders of magnitude, which has enabled in vivo metabolic imaging by MRI. However, because of the short lifetime of the hyperpolarized magnetization (typically <1 min), the polarization process must be carried out close to the point of use. Here we introduce a concept that markedly extends hyperpolarization lifetimes and enables the transportation of hyperpolarized metabolites. The hyperpolarized sample can thus be removed from the polarizer and stored or transported for use at remote MRI or NMR sites. We show that hyperpolarization in alanine and glycine survives 16 h storage and transport, maintaining overall polarization enhancements of up to three orders of magnitude. PMID:28072398

  9. NMR Hyperpolarization Techniques for Biomedicine

    PubMed Central

    Nikolaou, Panayiotis; Goodson, Boyd M.

    2015-01-01

    Recent developments in NMR hyperpolarization have enabled a wide array of new in vivo molecular imaging modalities—ranging from functional imaging of the lungs to metabolic imaging of cancer. This Concept article explores selected advances in methods for the preparation and use of hyperpolarized contrast agents, many of which are already at or near the phase of their clinical validation in patients. PMID:25470566

  10. Some electrical properties of the endothelium-dependent hyperpolarization recorded from rat arterial smooth muscle cells.

    PubMed Central

    Chen, G; Suzuki, H

    1989-01-01

    1. Electrical responses produced by acetylcholine (ACh) and histamine were recorded from smooth muscle cells of the intralobular small pulmonary artery (SPA), main pulmonary artery (MPA) and thoracic aorta of rats. 2. In MPA and SPA, ACh and histamine produced a transient hyperpolarization of the membrane, and the potential decayed exponentially with a time constant of 2-3 min. In aorta, ACh produced a sustained and histamine produced a transient hyperpolarization. 3. The ACh- and histamine-induced hyperpolarizations were blocked by atropine and mepyramine, respectively, or by removing the endothelial cells. 4. The amplitude of the hyperpolarization was increased in low [K+]o solutions and decreased in high [K+]o solutions. The ionic conductance of the membrane was increased during the hyperpolarization, suggesting an involvement of the increased potassium conductance. 5. A reproducible amplitude of hyperpolarization was generated when ACh or histamine was applied at intervals of over 10 or 30 min, respectively. 6. In aorta, after the transient hyperpolarization had ceased during continued application of histamine, ACh again produced a hyperpolarization, i.e. the transient nature of the hyperpolarization was not due to desensitization of the receptor upon which the hyperpolarizing substance acted, assuming histamine and ACh release the same hyperpolarizing substance. 7. ACh and histamine relaxed the tissues from SPA, MPA and aorta during the noradrenaline (NA)- or high [K+]o solution-induced contraction, in a concentration-dependent manner, only when the endothelial cells were intact. Both ACh and histamine were potent relaxants in MPA and aorta, but showed weak relaxing actions in SPA. 8. In aorta, ACh and histamine produced a sustained relaxation for up to 10 min, and Methylene Blue diminished and altered it to a transient relaxation (for histamine) or an initial large, followed by a small sustained (for ACh), relaxation. 9. In the presence of NA and NA plus

  11. A magnetic tunnel to shelter hyperpolarized fluids

    SciTech Connect

    Milani, Jonas Vuichoud, Basile; Bornet, Aurélien; Miéville, Pascal; Mottier, Roger; Jannin, Sami; Bodenhausen, Geoffrey

    2015-02-15

    To shield solutions carrying hyperpolarized nuclear magnetization from rapid relaxation during transfer through low fields, the transfer duct can be threaded through an array of permanent magnets. The advantages are illustrated for solutions containing hyperpolarized {sup 1}H and {sup 13}C nuclei in a variety of molecules.

  12. Membrane hyperpolarization during human sperm capacitation

    PubMed Central

    López-González, I.; Torres-Rodríguez, P.; Sánchez-Carranza, O.; Solís-López, A.; Santi, C.M.; Darszon, A.; Treviño, C.L.

    2014-01-01

    Sperm capacitation is a complex and indispensable physiological process that spermatozoa must undergo in order to acquire fertilization capability. Spermatozoa from several mammalian species, including mice, exhibit a capacitation-associated plasma membrane hyperpolarization, which is necessary for the acrosome reaction to occur. Despite its importance, this hyperpolarization event has not been adequately examined in human sperm. In this report we used flow cytometry to show that a subpopulation of human sperm indeed undergo a plasma membrane hyperpolarization upon in vitro capacitation. This hyperpolarization correlated with two other well-characterized capacitation parameters, namely an increase in intracellular pH and Ca2+ concentration, measured also by flow cytometry. We found that sperm membrane hyperpolarization was completely abolished in the presence of a high external K+ concentration (60 mM), indicating the participation of K+ channels. In order to identify, which of the potential K+ channels were involved in this hyperpolarization, we used different K+ channel inhibitors including charybdotoxin, slotoxin and iberiotoxin (which target Slo1) and clofilium (a more specific blocker for Slo3). All these K+ channel antagonists inhibited membrane hyperpolarization to a similar extent, suggesting that both members of the Slo family may potentially participate. Two very recent papers recorded K+ currents in human sperm electrophysiologically, with some contradictory results. In the present work, we show through immunoblotting that Slo3 channels are present in the human sperm membrane. In addition, we found that human Slo3 channels expressed in CHO cells were sensitive to clofilium (50 μM). Considered altogether, our data indicate that Slo1 and Slo3 could share the preponderant role in the capacitation-associated hyperpolarization of human sperm in contrast to what has been previously reported for mouse sperm, where Slo3 channels are the main contributors to the

  13. Hyperpolarized (129)Xe T (1) in oxygenated and deoxygenated blood

    NASA Technical Reports Server (NTRS)

    Albert, M. S.; Balamore, D.; Kacher, D. F.; Venkatesh, A. K.; Jolesz, F. A.

    2000-01-01

    The viability of the new technique of hyperpolarized (129)Xe MRI (HypX-MRI) for imaging organs other than the lungs depends on whether the spin-lattice relaxation time, T(1), of (129)Xe is sufficiently long in the blood. In previous experiments by the authors, the T(1) was found to be strongly dependent upon the oxygenation of the blood, with T(1) increasing from about 3 s in deoxygenated samples to about 10 s in oxygenated samples. Contrarily, Tseng et al. (J. Magn. Reson. 1997; 126: 79-86) reported extremely long T(1) values deduced from an indirect experiment in which hyperpolarized (129)Xe was used to create a 'blood-foam'. They found that oxygenation decreased T(1). Pivotal to their experiment is the continual and rapid exchange of hyperpolarized (129)Xe between the gas phase (within blood-foam bubbles) and the dissolved phase (in the skin of the bubbles); this necessitated a complicated analysis to extract the T(1) of (129)Xe in blood. In the present study, the experimental design minimizes gas exchange after the initial bolus of hyperpolarized (129)Xe has been bubbled through the sample. This study confirms that oxygenation increases the T(1) of (129)Xe in blood, from about 4 s in freshly drawn venous blood, to about 13 s in blood oxygenated to arterial levels, and also shifts the red blood cell resonance to higher frequency. Copyright 2000 John Wiley & Sons, Ltd. Abbreviations used BOLD blood oxygen level dependent NOE nuclear overhouses effect PO(2) oxygen partial pressure RBC red blood cells RF radio frequency SNR signal-to-noise ratio.

  14. Hyperpolarized 131Xe NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Stupic, Karl F.; Cleveland, Zackary I.; Pavlovskaya, Galina E.; Meersmann, Thomas

    2011-01-01

    Hyperpolarized (hp) 131Xe with up to 2.2% spin polarization (i.e., 5000-fold signal enhancement at 9.4 T) was obtained after separation from the rubidium vapor of the spin-exchange optical pumping (SEOP) process. The SEOP was applied for several minutes in a stopped-flow mode, and the fast, quadrupolar-driven T1 relaxation of this spin I = 3/2 noble gas isotope required a rapid subsequent rubidium removal and swift transfer into the high magnetic field region for NMR detection. Because of the xenon density dependent 131Xe quadrupolar relaxation in the gas phase, the SEOP polarization build-up exhibits an even more pronounced dependence on xenon partial pressure than that observed in 129Xe SEOP. 131Xe is the only stable noble gas isotope with a positive gyromagnetic ratio and shows therefore a different relative phase between hp signal and thermal signal compared to all other noble gases. The gas phase 131Xe NMR spectrum displays a surface and magnetic field dependent quadrupolar splitting that was found to have additional gas pressure and gas composition dependence. The splitting was reduced by the presence of water vapor that presumably influences xenon-surface interactions. The hp 131Xe spectrum shows differential line broadening, suggesting the presence of strong adsorption sites. Beyond hp 131Xe NMR spectroscopy studies, a general equation for the high temperature, thermal spin polarization, P, for spin I⩾1/2 nuclei is presented.

  15. Hyperpolarized 131Xe NMR spectroscopy

    PubMed Central

    Stupic, Karl F.; Cleveland, Zackary I.; Pavlovskaya, Galina E.; Meersmann, Thomas

    2011-01-01

    Hyperpolarized (hp) 131Xe with up to 2.2% spin polarization (i.e., 5000-fold signal enhancement at 9.4 T) was obtained after separation from the rubidium vapor of the spin-exchange optical pumping (SEOP) process. The SEOP was applied for several minutes in a stopped-flow mode, and the fast, quadrupolar-driven T1 relaxation of this spin I = 3/2 noble gas isotope required a rapid subsequent rubidium removal and swift transfer into the high magnetic field region for NMR detection. Because of the xenon density dependent 131Xe quadrupolar relaxation in the gas phase, the SEOP polarization build-up exhibits an even more pronounced dependence on xenon partial pressure than that observed in 129Xe SEOP. 131Xe is the only stable noble gas isotope with a positive gyromagnetic ratio and shows therefore a different relative phase between hp signal and thermal signal compared to all other noble gases. The gas phase 131Xe NMR spectrum displays a surface and magnetic field dependent quadrupolar splitting that was found to have additional gas pressure and gas composition dependence. The splitting was reduced by the presence of water vapor that presumably influences xenon-surface interactions. The hp 131Xe spectrum shows differential line broadening, suggesting the presence of strong adsorption sites. Beyond hp 131Xe NMR spectroscopy studies, a general equation for the high temperature, thermal spin polarization, P, for spin I⩾1/2 nuclei is presented. PMID:21051249

  16. Hyperpolarized NMR Probes for Biological Assays

    PubMed Central

    Meier, Sebastian; Jensen, Pernille R.; Karlsson, Magnus; Lerche, Mathilde H.

    2014-01-01

    During the last decade, the development of nuclear spin polarization enhanced (hyperpolarized) molecular probes has opened up new opportunities for studying the inner workings of living cells in real time. The hyperpolarized probes are produced ex situ, introduced into biological systems and detected with high sensitivity and contrast against background signals using high resolution NMR spectroscopy. A variety of natural, derivatized and designed hyperpolarized probes has emerged for diverse biological studies including assays of intracellular reaction progression, pathway kinetics, probe uptake and export, pH, redox state, reactive oxygen species, ion concentrations, drug efficacy or oncogenic signaling. These probes are readily used directly under natural conditions in biofluids and are often directly developed and optimized for cellular assays, thus leaving little doubt about their specificity and utility under biologically relevant conditions. Hyperpolarized molecular probes for biological NMR spectroscopy enable the unbiased detection of complex processes by virtue of the high spectral resolution, structural specificity and quantifiability of NMR signals. Here, we provide a survey of strategies used for the selection, design and use of hyperpolarized NMR probes in biological assays, and describe current limitations and developments. PMID:24441771

  17. Do twisted laser beams evoke nuclear hyperpolarization?

    NASA Astrophysics Data System (ADS)

    Schmidt, A. B.; Andrews, D. L.; Rohrbach, A.; Gohn-Kreuz, C.; Shatokhin, V. N.; Kiselev, V. G.; Hennig, J.; von Elverfeldt, D.; Hövener, J.-B.

    2016-07-01

    The hyperpolarization of nuclear spins promises great advances in chemical analysis and medical diagnosis by substantially increasing the sensitivity of nuclear magnetic resonance (NMR). Current methods to produce a hyperpolarized sample, however, are arduous, time-consuming or costly and require elaborate equipment. Recently, a much simpler approach was introduced that holds the potential, if harnessed appropriately, to revolutionize the production of hyperpolarized spins. It was reported that high levels of hyperpolarization in nuclear spins can be created by irradiation with a laser beam carrying orbital angular momentum (twisted light). Aside from these initial reports however, no further experimental verification has been presented. In addition, this effect has so far evaded a critical theoretical examination. In this contribution, we present the first independent attempt to reproduce the effect. We exposed a sample of immersion oil or a fluorocarbon liquid that was placed within a low-field NMR spectrometer to Laguerre-Gaussian and Bessel laser beams at a wavelength of 514.5 nm and various topological charges. We acquired 1H and 19F NMR free induction decay data, either during or alternating with the irradiation that was parallel to B0. We observed an irregular increase in NMR signal in experiments where the sample was exposed to beams with higher values of the topological charge. However, at no time did the effect reach statistical significance of 95%. Given the measured sensitivity of our setup, we estimate that a possible effect did not exceed a hyperpolarization (at 5 mT) of 0.14-6%, depending on the assumed hyperpolarized volume. It should be noted though, that there were some differences between our setup and the previous implementation of the experiment, which may have inhibited the full incidence of this effect. To approach a theoretical description of this effect, we considered the interaction of an electron with a plane wave, which is known to be

  18. NMR and MRI of blood-dissolved hyperpolarized Xe-129 in different hollow-fiber membranes.

    PubMed

    Amor, Nadia; Hamilton, Kathrin; Küppers, Markus; Steinseifer, Ulrich; Appelt, Stephan; Blümich, Bernhard; Schmitz-Rode, Thomas

    2011-11-18

    Magnetic resonance of hyperpolarized (129)Xe has found a wide field of applications in the analysis of biologically relevant fluids. Recently, it has been shown that the dissolution of hyperpolarized gas into the fluid via hollow-fiber membranes leads to bubble-free (129)Xe augmentation, and thus to an enhanced signal. In addition, hollow-fiber membranes permit a continuous operation mode. Herein, a quantitative magnetic resonance imaging and spectroscopy analysis of a customized hollow-fiber membrane module is presented. Different commercial hollow-fiber membrane types are compared with regard to their (129)Xe dissolution efficiency into porcine blood, its constituents, and other fluids. The presented study gives new insight into the suitability of these hollow-fiber membrane types for hyperpolarized gas dissolution setups.

  19. Hyperpolarized Gas MRI: Technique and Applications

    PubMed Central

    McAdams, Holman P.; Kaushik, S. Sivaram; Driehuys, Bastiaan

    2015-01-01

    Synopsis Functional imaging today offers a rich world of information that is more sensitive to changes in lung structure and function than traditionally obtained pulmonary function tests. Hyperpolarized helium (3He) and xenon (129Xe) MR imaging of the lungs provided new sensitive contrast mechanisms to probe changes in pulmonary ventilation, microstructure and gas exchange. With the recent scarcity in the supply of 3He the field of hyperpolarized gas imaging shifted to the use of cheaper and naturally available 129Xe. Xenon is well tolerated and recent technical advances have ensured that the 129Xe image quality is on par with that of 3He. The added advantage of 129Xe is its solubility in pulmonary tissue, which allows exploring specific lung function characteristics involved in gas exchange and alveolar oxygenation. With a plethora of contrast mechanisms, hyperpolarized gases and 129Xe in particular, stands to be an excellent probe of pulmonary structure and function, and provide sensitive and non-invasive biomarkers for a wide variety of pulmonary diseases. PMID:25952516

  20. Transmembrane exchange of hyperpolarized 13C-urea in human erythrocytes: subminute timescale kinetic analysis.

    PubMed

    Pagès, Guilhem; Puckeridge, Max; Liangfeng, Guo; Tan, Yee Ling; Jacob, Chacko; Garland, Marc; Kuchel, Philip W

    2013-11-05

    The rate of exchange of urea across the membranes of human erythrocytes (red blood cells) was quantified on the 1-s to 2-min timescale. (13)C-urea was hyperpolarized and subjected to rapid dissolution and the previously reported (partial) resolution of (13)C NMR resonances from the molecules inside and outside red blood cells in suspensions was observed. This enabled a stopped-flow type of experiment to measure the (initially) zero-trans transport of urea with sequential single-pulse (13)C NMR spectra, every second for up to ~2 min. Data were analyzed using Bayesian reasoning and a Markov chain Monte Carlo method with a set of simultaneous nonlinear differential equations that described nuclear magnetic relaxation combined with transmembrane exchange. Our results contribute to quantitative understanding of urea-exchange kinetics in the whole body; and the methodological approach is likely to be applicable to other cellular systems and tissues in vivo.

  1. Hyperpolarized water as an authentic magnetic resonance imaging contrast agent

    PubMed Central

    McCarney, Evan R.; Armstrong, Brandon D.; Lingwood, Mark D.; Han, Songi

    2007-01-01

    Pure water in a highly 1H spin-polarized state is proposed as a contrast-agent-free contrast agent to visualize its macroscopic evolution in aqueous media by MRI. Remotely enhanced liquids for image contrast (RELIC) utilizes a 1H signal of water that is enhanced outside the sample in continuous-flow mode and immediately delivered to the sample to obtain maximum contrast between entering and bulk fluids. Hyperpolarization suggests an ideal contrast mechanism to highlight the ubiquitous and specific function of water in physiology, biology, and materials because the physiological, chemical, and macroscopic function of water is not altered by the degree of magnetization. We present an approach that is capable of instantaneously enhancing the 1H MRI signal by up to 2 orders of magnitude through the Overhauser effect under ambient conditions at 0.35 tesla by using highly spin-polarized unpaired electrons that are covalently immobilized onto a porous, water-saturated gel matrix. The continuous polarization of radical-free flowing water allowed us to distinctively visualize vortices in model reactors and dispersion patterns through porous media. A 1H signal enhancement of water by a factor of −10 and −100 provides for an observation time of >4 and 7 s, respectively, upon its injection into fluids with a T1 relaxation time of >1.5 s. The implications for chemical engineering or biomedical applications of using hyperpolarized solvents or physiological fluids to visualize mass transport and perfusion with high and authentic MRI contrast originating from water itself, and not from foreign contrast agents, are immediate. PMID:17264210

  2. Perfusion Imaging with a Freely Diffusible Hyperpolarized Contrast Agent

    PubMed Central

    Grant, Aaron K.; Vinogradov, Elena; Wang, Xiaoen; Lenkinski, Robert E.; Alsop, David C.

    2011-01-01

    Contrast agents that can diffuse freely into or within tissue have numerous attractive features for perfusion imaging. Here we present preliminary data illustrating the suitability of hyperpolarized 13C labeled 2-methylpropan-2-ol (also known as dimethylethanol, tertiary butyl alcohol and tert-butanol) as a freely diffusible contrast agent for magnetic resonance perfusion imaging. Dynamic 13C images acquired in rat brain with a balanced steady-state free precession (bSSFP) sequence following administration of hyperpolarized 2-methylpropan-2-ol show that this agent can be imaged with 2–4s temporal resolution, 2mm slice thickness, and 700 micron in-plane resolution while retaining adequate signal-to-noise ratio. 13C relaxation measurements on 2-methylpropan-2-ol in blood at 9.4T yield T1=46±4s and T2=0.55±0.03s. In the rat brain at 4.7T, analysis of the temporal dynamics of the bSSFP image intensity in tissue and venous blood indicate that 2-methylpropan-2-ol has a T2 of roughly 2–4s and a T1 of 43±24s. In addition, the images indicate that 2-methylpropan-2-ol is freely diffusible in brain and hence has a long residence time in tissue; this in turn makes it possible to image the agent continuously for tens of seconds. These characteristics show that 2-methylpropan-2-ol is a promising agent for robust and quantitative perfusion imaging in the brain and body. PMID:21432901

  3. Hyperpolarized 13C MR Markers of Renal Tumor Aggressiveness

    DTIC Science & Technology

    2013-10-01

    reliably distinguish renal cancer aggressiveness for optimal triage of therapies . Hyperpolarized (HP) 13C magnetic resonance spectroscopic imaging (MRSI...reliably distinguish renal cancer aggressiveness for optimal triage of therapies . Hyperpolarized (HP) 13C magnetic resonance spectroscopic imaging (MRSI) is... cancer and normal tissues were obtained from nephrectomy specimens and sliced using Krumdieck slicer. With a precision gauge micrometer, the slice

  4. Hyperpolarized and inert gas MRI: the future.

    PubMed

    Couch, Marcus J; Blasiak, Barbara; Tomanek, Boguslaw; Ouriadov, Alexei V; Fox, Matthew S; Dowhos, Krista M; Albert, Mitchell S

    2015-04-01

    Magnetic resonance imaging (MRI) is a potentially ideal imaging modality for noninvasive, nonionizing, and longitudinal assessment of disease. Hyperpolarized (HP) agents have been developed in the past 20 years for MR imaging, and they have the potential to vastly improve MRI sensitivity for the diagnosis and management of various diseases. The polarization of nuclear magnetic resonance (NMR)-sensitive nuclei other than (1)H (e.g., (3)He, (129)Xe) can be enhanced by a factor of up to 100,000 times above thermal equilibrium levels, which enables direct detection of the HP agent with no background signal. In this review, a number of HP media applications in MR imaging are discussed, including HP (3)He and (129)Xe lung imaging, HP (129)Xe brain imaging, and HP (129)Xe biosensors. Inert fluorinated gas MRI, which is a new lung imaging technique that does not require hyperpolarization, is also briefly discussed. This technique will likely be an important future direction for the HP gas lung imaging community.

  5. Silicon Nanoparticles as Hyperpolarized Magnetic Resonance Imaging Agents

    PubMed Central

    Aptekar, Jacob W.; Cassidy, Maja C.; Johnson, Alexander C.; Barton, Robert A.; Lee, Menyoung; Ogier, Alexander C.; Vo, Chinh; Anahtar, Melis N.; Ren, Yin; Bhatia, Sangeeta N.; Ramanathan, Chandrasekhar; Cory, David G.; Hill, Alison L.; Mair, Ross W.; Rosen, Matthew S.; Walsworth, Ronald L.

    2014-01-01

    Magnetic resonance imaging of hyperpolarized nuclei provides high image contrast with little or no background signal. To date, in-vivo applications of pre-hyperpolarized materials have been limited by relatively short nuclear spin relaxation times. Here, we investigate silicon nanoparticles as a new type of hyperpolarized magnetic resonance imaging agent. Nuclear spin relaxation times for a variety of Si nanoparticles are found to be remarkably long, ranging from many minutes to hours at room temperature, allowing hyperpolarized nanoparticles to be transported, administered, and imaged on practical time scales. Additionally, we demonstrate that Si nanoparticles can be surface functionalized using techniques common to other biologically targeted nanoparticle systems. These results suggest that Si nanoparticles can be used as a targetable, hyperpolarized magnetic resonance imaging agent with a large range of potential applications. PMID:19950973

  6. 15N Hyperpolarization by Reversible Exchange Using SABRE-SHEATH

    PubMed Central

    2016-01-01

    NMR signal amplification by reversible exchange (SABRE) is a NMR hyperpolarization technique that enables nuclear spin polarization enhancement of molecules via concurrent chemical exchange of a target substrate and parahydrogen (the source of spin order) on an iridium catalyst. Recently, we demonstrated that conducting SABRE in microtesla fields provided by a magnetic shield enables up to 10% 15N-polarization (Theis, T.; et al. J. Am. Chem. Soc.2015, 137, 1404). Hyperpolarization on 15N (and heteronuclei in general) may be advantageous because of the long-lived nature of the hyperpolarization on 15N relative to the short-lived hyperpolarization of protons conventionally hyperpolarized by SABRE, in addition to wider chemical shift dispersion and absence of background signal. Here we show that these unprecedented polarization levels enable 15N magnetic resonance imaging. We also present a theoretical model for the hyperpolarization transfer to heteronuclei, and detail key parameters that should be optimized for efficient 15N-hyperpolarization. The effects of parahydrogen pressure, flow rate, sample temperature, catalyst-to-substrate ratio, relaxation time (T1), and reversible oxygen quenching are studied on a test system of 15N-pyridine in methanol-d4. Moreover, we demonstrate the first proof-of-principle 13C-hyperpolarization using this method. This simple hyperpolarization scheme only requires access to parahydrogen and a magnetic shield, and it provides large enough signal gains to enable one of the first 15N images (2 × 2 mm2 resolution). Importantly, this method enables hyperpolarization of molecular sites with NMR T1 relaxation times suitable for biomedical imaging and spectroscopy. PMID:25960823

  7. Sodium–calcium exchanger contributes to membrane hyperpolarization of intact endothelial cells from rat aorta during acetylcholine stimulation

    PubMed Central

    Bondarenko, Alexander

    2004-01-01

    The role of sodium–calcium exchanger in acetylcholine (Ach)-induced hyperpolarization of intact endothelial cells was studied in excised rat aorta. The membrane potential was recorded using perforated patch-clamp technique. The mean resting potential of endothelial cells was −44.1±1.4 mV. A selective inhibitor of sodium–calcium exchanger benzamil (100 μM) had no significant effect on resting membrane potential, but reversibly decreased the amplitude of sustained Ach-induced endothelial hyperpolarization from 20.9±1.4 to 5.7±1.1 mV when applied during the plateau phase. The blocker of reversed mode of the exchanger KB-R7943 (2-[2-[4-(4-nitrobenzyloxy)phenyl]ethyl]isothiourea methanesulfonate, 20 μM) reversibly decreased the amplitude of sustained Ach-induced hyperpolarization from 20.5±2.9 to 7.5±1.8 mV. Introduction of tetraethylammonium (10 mM) in the continuous presence of Ach decreased the sustained phase of hyperpolarization from 17.9±1.5 by 12.9±0.9 mV. Subsequent addition of 20 μM KB-R7943 further depolarized endothelial cells by 4.8±1.1 mV. Substituting external sodium with N-methyl D-glucamine during the plateau phase of Ach-evoked hyperpolarization reversibly decreased the hyperpolarization from −61.8±2.7 to −54.2±1.9 mV. In the majority of preparations, the initial response to removal of external sodium was a transient further rise in the membrane potential of several mV. Sodium ionophore monensin hyperpolarized endothelium by 10.3±0.7 mV. The inhibitory effect of benzamil on Ach-induced endothelial sustained hyperpolarization was observed in endothelium mechanically isolated from smooth muscle. These results suggest that the sodium–calcium exchanger of intact endothelial cells is able to operate in reverse following stimulation by Ach, contributing to sustained hyperpolarization. Myoendothelial electrical communications do not mediate the effect of blockers of sodium–calcium exchanger. PMID:15289290

  8. Functional lung imaging using hyperpolarized gas MRI.

    PubMed

    Fain, Sean B; Korosec, Frank R; Holmes, James H; O'Halloran, Rafael; Sorkness, Ronald L; Grist, Thomas M

    2007-05-01

    The noninvasive assessment of lung function using imaging is increasingly of interest for the study of lung diseases, including chronic obstructive pulmonary disease (COPD) and asthma. Hyperpolarized gas MRI (HP MRI) has demonstrated the ability to detect changes in ventilation, perfusion, and lung microstructure that appear to be associated with both normal lung development and disease progression. The physical characteristics of HP gases and their application to MRI are presented with an emphasis on current applications. Clinical investigations using HP MRI to study asthma, COPD, cystic fibrosis, pediatric chronic lung disease, and lung transplant are reviewed. Recent advances in polarization, pulse sequence development for imaging with Xe-129, and prototype low magnetic field systems dedicated to lung imaging are highlighted as areas of future development for this rapidly evolving technology.

  9. Biomedical imaging with hyperpolarized noble gases

    NASA Astrophysics Data System (ADS)

    Ruppert, Kai

    2014-11-01

    Hyperpolarized noble gases (HNGs), polarized to approximately 50% or higher, have led to major advances in magnetic resonance (MR) imaging of porous structures and air-filled cavities in human subjects, particularly the lung. By boosting the available signal to a level about 100 000 times higher than that at thermal equilibrium, air spaces that would otherwise appear as signal voids in an MR image can be revealed for structural and functional assessments. This review discusses how HNG MR imaging differs from conventional proton MR imaging, how MR pulse sequence design is affected and how the properties of gas imaging can be exploited to obtain hitherto inaccessible information in humans and animals. Current and possible future imaging techniques, and their application in the assessment of normal lung function as well as certain lung diseases, are described.

  10. Optical hyperpolarization of nitrogen donor spins in bulk diamond

    NASA Astrophysics Data System (ADS)

    Loretz, M.; Takahashi, H.; Segawa, T. F.; Boss, J. M.; Degen, C. L.

    2017-02-01

    We report hyperpolarization of the electronic spins associated with substitutional nitrogen defects in bulk diamond crystals. Hyperpolarization is achieved by optical pumping of nitrogen vacancy centers followed by rapid cross relaxation at the energy level matching condition in a 51 mT bias field. The maximum observed donor spin polarization is 0.9 % , corresponding to an enhancement of 25 compared to the thermal Boltzmann polarization. A further accumulation of polarization is impeded by an anomalous optical saturation effect that we attribute to charge state conversion processes. Hyperpolarized nitrogen donors may form a useful resource for increasing the efficiency of diamond-based dynamic nuclear polarization devices.

  11. LIGHT-SABRE enables efficient in-magnet catalytic hyperpolarization

    NASA Astrophysics Data System (ADS)

    Theis, Thomas; Truong, Milton; Coffey, Aaron M.; Chekmenev, Eduard Y.; Warren, Warren S.

    2014-11-01

    Nuclear spin hyperpolarization overcomes the sensitivity limitations of traditional NMR and MRI, but the most general method demonstrated to date (dynamic nuclear polarization) has significant limitations in scalability, cost, and complex apparatus design. As an alternative, signal amplification by reversible exchange (SABRE) of parahydrogen on transition metal catalysts can hyperpolarize a variety of substrates, but to date this scheme has required transfer of the sample to low magnetic field or very strong RF irradiation. Here we demonstrate "Low-Irradiation Generation of High Tesla-SABRE" (LIGHT-SABRE) which works with simple pulse sequences and low power deposition; it should be usable at any magnetic field and for hyperpolarization of many different nuclei. This approach could drastically reduce the cost and complexity of producing hyperpolarized molecules.

  12. Hyperpolarized nanodiamond with long spin-relaxation times

    PubMed Central

    Rej, Ewa; Gaebel, Torsten; Boele, Thomas; Waddington, David E.J.; Reilly, David J.

    2015-01-01

    The use of hyperpolarized agents in magnetic resonance, such as 13C-labelled compounds, enables powerful new imaging and detection modalities that stem from a 10,000-fold boost in signal. A major challenge for the future of the hyperpolarization technique is the inherently short spin-relaxation times, typically <60 s for 13C liquid-state compounds, which limit the time that the signal remains boosted. Here we demonstrate that 1.1% natural abundance 13C spins in synthetic nanodiamond can be hyperpolarized at cryogenic and room temperature without the use of free radicals, and, owing to their solid-state environment, exhibit relaxation times exceeding 1 h. Combined with the already established applications of nanodiamonds in the life sciences as inexpensive fluorescent markers and non-cytotoxic substrates for gene and drug delivery, these results extend the theranostic capabilities of nanoscale diamonds into the domain of hyperpolarized magnetic resonance. PMID:26450570

  13. Hyperpolarized nanodiamond with long spin-relaxation times

    NASA Astrophysics Data System (ADS)

    Rej, Ewa; Gaebel, Torsten; Boele, Thomas; Waddington, David E. J.; Reilly, David J.

    2015-10-01

    The use of hyperpolarized agents in magnetic resonance, such as 13C-labelled compounds, enables powerful new imaging and detection modalities that stem from a 10,000-fold boost in signal. A major challenge for the future of the hyperpolarization technique is the inherently short spin-relaxation times, typically <60 s for 13C liquid-state compounds, which limit the time that the signal remains boosted. Here we demonstrate that 1.1% natural abundance 13C spins in synthetic nanodiamond can be hyperpolarized at cryogenic and room temperature without the use of free radicals, and, owing to their solid-state environment, exhibit relaxation times exceeding 1 h. Combined with the already established applications of nanodiamonds in the life sciences as inexpensive fluorescent markers and non-cytotoxic substrates for gene and drug delivery, these results extend the theranostic capabilities of nanoscale diamonds into the domain of hyperpolarized magnetic resonance.

  14. Towards hyperpolarized 13C-succinate imaging of brain cancer

    NASA Astrophysics Data System (ADS)

    Bhattacharya, Pratip; Chekmenev, Eduard Y.; Perman, William H.; Harris, Kent C.; Lin, Alexander P.; Norton, Valerie A.; Tan, Chou T.; Ross, Brian D.; Weitekamp, Daniel P.

    2007-05-01

    We describe a novel 13C enriched precursor molecule, sodium 1- 13C acetylenedicarboxylate, which after hydrogenation by PASADENA (Parahydrogen and Synthesis Allows Dramatically Enhanced Nuclear Alignment) under controlled experimental conditions, becomes hyperpolarized 13C sodium succinate. Fast in vivo 3D FIESTA MR imaging demonstrated that, following carotid arterial injection, the hyperpolarized 13C-succinate appeared in the head and cerebral circulation of normal and tumor-bearing rats. At this time, no in vivo hyperpolarized signal has been localized to normal brain or brain tumor. On the other hand, ex vivo samples of brain harvested from rats bearing a 9L brain tumor, 1 h or more following in vivo carotid injection of hyperpolarized 13C sodium succinate, contained significant concentrations of the injected substrate, 13C sodium succinate, together with 13C maleate and succinate metabolites 1- 13C-glutamate, 5- 13C-glutamate, 1- 13C-glutamine and 5- 13C-glutamine. The 13C substrates and products were below the limits of NMR detection in ex vivo samples of normal brain consistent with an intact blood-brain barrier. These ex vivo results indicate that hyperpolarized 13C sodium succinate may become a useful tool for rapid in vivo identification of brain tumors, providing novel biomarkers in 13C MR spectral-spatial images.

  15. A hyperpolarized choline molecular probe for monitoring acetylcholine synthesis.

    PubMed

    Allouche-Arnon, Hyla; Gamliel, Ayelet; Barzilay, Claudia M; Nalbandian, Ruppen; Gomori, J Moshe; Karlsson, Magnus; Lerche, Mathilde H; Katz-Brull, Rachel

    2011-01-01

    Choline as a reporter molecule has been investigated by in vivo magnetic resonance for almost three decades. Accumulation of choline metabolites (mainly the phosphorylated forms) had been observed in malignancy in preclinical models, ex-vivo, in vivo and in patients. The combined choline metabolite signal appears in (1) H-MRS of the brain and its relative intensity had been used as a diagnostic factor in various conditions. The advent of spin hyperpolarization methods for in vivo use has raised interest in the ability to follow the physiological metabolism of choline into acetylcholine in the brain. Here we present a stable-isotope labeled choline analog, [1,1,2,2-D(4) ,2-(13) C]choline chloride, that is suitable for this purpose. In this analog, the (13) C position showed 24% polarization in the liquid state, following DNP hyperpolarization. This nucleus also showed a long T(1) (35 s) at 11.8 T and 25 °C, which is a prerequisite for hyperpolarized studies. The chemical shift of this (13) C position differentiates choline and acetylcholine from each other and from the other water-soluble choline metabolites, namely phosphocholine and betaine. Enzymatic studies using an acetyltransferase enzyme showed the synthesis of the deuterated-acetylcholine form at thermal equilibrium conditions and in a hyperpolarized state. Analysis using a comprehensive model showed that the T(1) of the formed hyperpolarized [1,1,2,2-D(4) ,2-(13) C]acetylcholine was 34 s at 14.1 T and 37 °C. We conclude that [1,1,2,2-D(4) ,2-(13) C]choline chloride is a promising new molecular probe for hyperpolarized metabolic studies and discuss the factors related to its possible use in vivo.

  16. PASADENA hyperpolarization of 13C biomolecules: equipment design and installation

    PubMed Central

    Hövener, Jan-Bernd; Chekmenev, Eduard Y.; Harris, Kent C.; Perman, William H.; Robertson, Larry W.; Bhattacharya, Pratip

    2009-01-01

    Object The PASADENA method has achieved hyperpolarization of 16–20% (exceeding 40,000-fold signal enhancement at 4.7 T), in liquid samples of biological molecules relevant to in vivo MRI and MRS. However, there exists no commercial apparatus to perform this experiment conveniently and reproducibly on the routine basis necessary for translation of PASADENA to questions of biomedical importance. The present paper describes equipment designed for rapid production of six to eight liquid samples per hour with high reproducibility of hyperpolarization. Materials and methods Drawing on an earlier, but unpublished, prototype, we provide diagrams of a delivery circuit, a laminar-flow reaction chamber within a low field NMR contained in a compact, movable housing. Assembly instructions are provided from which a computer driven, semiautomated PASADENA polarizer can be constructed. Results Together with an available parahydrogen generator, the polarizer, which can be operated by a single investigator, completes one cycle of hyperpolarization each 52 s. Evidence of efficacy is presented. In contrast to competing, commercially available devices for dynamic nuclear polarization which characteristically require 90 min per cycle, PASADENA provides a low-cost alternative for high throughput. Conclusions This equipment is suited to investigators who have an established small animal NMR and wish to explore the potential of heteronuclear (13C and 15N) MRI, MRS, which harnesses the enormous sensitivity gain offered by hyperpolarization. PMID:19067008

  17. Large Production of Hyperpolarized 129-Xe for MRI Applications

    NASA Astrophysics Data System (ADS)

    Ruset, Iulian; Hersman, F. W.; Distelbrink, Jan; Ketel, Stephen; Covrig, Silviu; Muradian, Iga; Sindile, Adrian

    2007-03-01

    Although 129-Xe was the first hyperpolarized gas to be used in MRI studies, the research community has focused on 3-He, mainly because of the larger quantities of hyperpolarized gas available. Xenon has advantages over helium, such as natural abundance, lower diffusion, and high solubility in blood. It presents a large frequency chemical shift when dissolved in blood, tissue, brain, or trapped in molecular cages. A new design of a high-flow low-pressure spin-exchange optical pumping Rb-Xe polarizer was recently demonstrated by our group. The concept of counterflowing the gas mixture against laser light and dividing the polarizing cell in three operational zones has resulted in an increase with over an order of magnitude in the output magnetization compared with previously reported polarizers. We were able to produce hyperpolarized xenon at 64% polarization for 0.3 liters/hour flow rate and 22% polarization at 6 liters/hour. We also demonstrated a new design of freezing and thawing hyperpolarized xenon with minimum losses. We will present the concept of the high-flow low-pressure counterflowing xenon polarizer, its performance, as well as new optical pumping laser technologies. We will discuss optimization plans for xenon polarizing systems based on experimental observed limitations and theoretical modeling.

  18. Probing alanine transaminase catalysis with hyperpolarized 13CD3-pyruvate

    PubMed Central

    Barb, A.W.; Hekmatyar, S.K.; Glushka, J.N.; Prestegard, J.H.

    2013-01-01

    Hyperpolarized metabolites offer a tremendous sensitivity advantage (>104 fold) when measuring flux and enzyme activity in living tissues by magnetic resonance methods. These sensitivity gains can also be applied to mechanistic studies that impose time and metabolite concentration limitations. Here we explore the use of hyperpolarization by dissolution dynamic nuclear polarization (DNP) in mechanistic studies of alanine transaminase (ALT), a well-established biomarker of liver disease and cancer that converts pyruvate to alanine using glutamate as a nitrogen donor. A specific deuterated, 13C-enriched analog of pyruvic acid, 13C3D3-pyruvic acid, is demonstrated to have advantages in terms of detection by both direct 13C observation and indirect observation through methyl protons introduced by ALT-catalyzed H–D exchange. Exchange on injecting hyperpolarized 13C3D3-pyruvate into ALT dissolved in buffered 1H2O, combined with an experimental approach to measure proton incorporation, provided information on mechanistic details of transaminase action on a 1.5 s timescale. ALT introduced, on average, 0.8 new protons into the methyl group of the alanine produced, indicating the presence of an off-pathway enamine intermediate. The opportunities for exploiting mechanism-dependent molecular signatures as well as indirect detection of hyperpolarized 13C3-pyruvate and products in imaging applications are discussed. PMID:23357427

  19. Responses of human sensory and motor axons to the release of ischaemia and to hyperpolarizing currents.

    PubMed

    Lin, Cindy S-Y; Kuwabara, Satoshi; Cappelen-Smith, Cecilia; Burke, David

    2002-06-15

    This study compared directly the post-ischaemic behaviour of sensory and motor axons in the human median nerve, focusing on the excitability changes produced by ischaemia and its release and by continuous polarizing DC. The decrease in threshold during ischaemia for 13 min was greater, the post-ischaemic increase in threshold was more rapid, and the return to the pre-ischaemic excitability took longer in sensory axons. However, a transient depolarizing threshold shift developed in sensory axons a few minutes after release of ischaemia. This pattern could not be reproduced by polarizing currents designed to mimic the probable pump-induced changes in membrane potential, even though the applied currents produced greater changes in threshold. Hyperpolarizing currents of equivalent intensity produced a greater increase in threshold for motor axons than sensory axons and, in studies of threshold electrotonus using graded hyperpolarizing DC, accommodation was greater in sensory than motor axons. The post-ischaemic changes in threshold were not uniform for axons of different threshold, whether sensory or motor, the threshold increase was usually less prominent for low-threshold axons. A transient post-ischaemic depolarization could be produced in motor axons with ischaemia of 20 min duration. Greater ischaemic and post-ischaemic changes in threshold for sensory axons could reflect greater dependence on the electrogenic Na+-K+ pump to maintain resting membrane potential and/or greater extracellular K+ accumulation in ischaemic sensory axons. Inward K+ currents due to extracellular K+ accumulation would then be more likely to trigger a depolarizing shift in membrane potential, the degree of K+ accumulation and pump activity being dependent on the duration of ischaemia. In sensory axons the greater tendency to accommodate to hyperpolarizing stimuli presumably contributes to shaping their post-ischaemic behaviour but is probably insufficient to explain why their behaviour

  20. Imaging pH with hyperpolarized 13C.

    PubMed

    Gallagher, Ferdia A; Kettunen, Mikko I; Brindle, Kevin M

    2011-10-01

    pH is a fundamental physiological parameter that is tightly controlled by endogenous buffers. The acid-base balance is altered in many disease states, such as inflammation, ischemia and cancer. Despite the importance of pH, there are currently no routine methods for imaging the spatial distribution of pH in humans. The enormous gain in sensitivity afforded by dynamic nuclear polarization (DNP) has provided a novel way in which to image tissue pH using MR, which has the potential to be translated into the clinic. This review explores the advantages and disadvantages of current pH imaging techniques and how they compare with DNP-based approaches for the measurement and imaging of pH with hyperpolarized (13)C. Intravenous injection of hyperpolarized (13)C-labeled bicarbonate results in the rapid production of hyperpolarized (13)CO(2) in the reaction catalyzed by carbonic anhydrase. As this reaction is close to equilibrium in the body and is pH dependent, the ratio of the (13)C signal intensities from H(13)CO(3)(-) and (13)CO(2), measured using MRS, can be used to calculate pH in vivo. The application of this technique to a murine tumor model demonstrated that it measured predominantly extracellular pH and could be mapped in the animal using spectroscopic imaging techniques. A second approach has been to use the production of hyperpolarized (13)CO(2) from hyperpolarized [1-(13)C]pyruvate to measure predominantly intracellular pH. In tissues with a high aerobic capacity, such as the heart, the hyperpolarized [1-(13)C]pyruvate undergoes rapid oxidative decarboxylation, catalyzed by intramitochondrial pyruvate dehydrogenase. Provided that there is sufficient carbonic anhydrase present to catalyze the rapid equilibration of the hyperpolarized (13)C label between CO(2) and bicarbonate, the ratio of their resonance intensities may again be used to estimate pH, which, in this case, is predominantly intracellular. As both pyruvate and bicarbonate are endogenous molecules they

  1. Propane-d6 Heterogeneously Hyperpolarized by Parahydrogen

    PubMed Central

    2015-01-01

    Long-lived spin states of hyperpolarized propane-d6 gas were demonstrated following pairwise addition of parahydrogen gas to propene-d6 using heterogeneous parahydrogen-induced polarization (HET-PHIP). Hyperpolarized molecules were synthesized using Rh/TiO2 solid catalyst with 1.6 nm Rh nanoparticles. Hyperpolarized (PH ∼ 1%) propane-d6 was detected at high magnetic field (9.4 T) spectroscopically and by high-resolution 3D gradient-echo MRI (4.7 T) as the gas flowed through the radiofrequency coil with a spatial and temporal resolution of 0.5 × 0.5 × 0.5 mm3 and 17.7 s, respectively. Stopped-flow hyperpolarized propane-d6 gas was also detected at 0.0475 T with an observed nuclear spin polarization of PH ∼ 0.1% and a relatively long lifetime with T1,eff = 6.0 ± 0.3 s. Importantly, it was shown that the hyperpolarized protons of the deuterated product obtained via pairwise parahydrogen addition could be detected directly at low magnetic field. Importantly, the relatively long low-field T1,eff of HP propane-d6 gas is not susceptible to paramagnetic impurities as tested by exposure to ∼0.2 atm oxygen. This long lifetime and nontoxic nature of propane gas could be useful for bioimaging applications including potentially pulmonary low-field MRI. The feasibility of high-resolution low-field 2D gradient-echo MRI was demonstrated with 0.88 × 0.88 mm2 spatial and ∼0.7 s temporal resolution, respectively, at 0.0475 T. PMID:25506406

  2. Chloride dependence of hyperpolarization-activated chloride channel gates

    PubMed Central

    Pusch, Michael; Jordt, Sven-Eric; Stein, Valentin; Jentsch, Thomas J

    1999-01-01

    ClC proteins are a class of voltage-dependent Cl− channels with several members mutated in human diseases. The prototype ClC-0 Torpedo channel is a dimeric protein; each subunit forms a pore that can gate independently from the other one. A common slower gating mechanism acts on both pores simultaneously; slow gating activates ClC-0 at hyperpolarized voltages. The ClC-2 Cl− channel is also activated by hyperpolarization, as are some ClC-1 mutants (e.g. D136G) and wild-type (WT) ClC-1 at certain pH values.We studied the dependence on internal Cl− ([Cl−]i) of the hyperpolarization-activated gates of several ClC channels (WT ClC-0, ClC-0 mutant P522G, ClC-1 mutant D136G and an N-terminal deletion mutant of ClC-2), by patch clamping channels expressed in Xenopus oocytes.With all these channels, reducing [Cl−]i shifted activation to more negative voltages and reduced the maximal activation at most negative voltages.We also investigated the external halide dependence of WT ClC-2 using two-electrode voltage-clamp recording. Reducing external Cl− ([Cl−]o) activated ClC-2 currents. Replacing [Cl−]o by the less permeant Br− reduced channel activity and accelerated deactivation.Gating of the ClC-2 mutant K566Q in normal [Cl−]o resembled that of WT ClC-2 in low [Cl−]o, i.e. channels had a considerable open probability (Po) at resting membrane potential. Substituting external Cl− by Br− or I− led to a decrease in Po.The [Cl−]i dependence of the hyperpolarization-activated gates of various ClC channels suggests a similar gating mechanism, and raises the possibility that the gating charge for the hyperpolarization-activated gate is provided by Cl−.The external halide dependence of hyperpolarization-activated gating of ClC-2 suggests that it is mediated or modulated by anions as in other ClC channels. In contrast to the depolarization-activated fast gates of ClC-0 and ClC-1, the absence of Cl− favours channel opening. Lysine 556 may be important

  3. Physiological response of rats to delivery of helium and xenon: implications for hyperpolarized noble gas imaging

    NASA Technical Reports Server (NTRS)

    Ramirez, M. P.; Sigaloff, K. C.; Kubatina, L. V.; Donahue, M. A.; Venkatesh, A. K.; Albert, M. S.; ALbert, M. S. (Principal Investigator)

    2000-01-01

    The physiological effects of various hyperpolarized helium and xenon MRI-compatible breathing protocols were investigated in 17 Sprague-Dawley rats, by continuous monitoring of blood oxygen saturation, heart rate, EKG, temperature and endotracheal pressure. The protocols included alternating breaths of pure noble gas and oxygen, continuous breaths of pure noble gas, breath-holds of pure noble gas for varying durations, and helium breath-holds preceded by two helium rinses. Alternate-breath protocols up to 128 breaths caused a decrease in oxygen saturation level of less than 5% for either helium or xenon, whereas 16 continuous-breaths caused a 31.5% +/- 2.3% decrease in oxygen saturation for helium and a 30.7% +/- 1. 3% decrease for xenon. Breath-hold protocols up to 25 s did not cause the oxygen saturation to fall below 90% for either of the noble gases. Oxygen saturation values below 90% are considered pathological. At 30 s of breath-hold, the blood oxygen saturation dropped precipitously to 82% +/- 0.6% for helium, and to 76.5% +/- 7. 4% for xenon. Breath-holds longer than 10 s preceded by pre-rinses caused oxygen saturation to drop below 90%. These findings demonstrate the need for standardized noble gas inhalation procedures that have been carefully tested, and for continuous physiological monitoring to ensure the safety of the subject. We find short breath-hold and alternate-breath protocols to be safe procedures for use in hyperpolarized noble gas MRI experiments. Copyright 2000 John Wiley & Sons, Ltd.

  4. SU-E-J-120: Comparing 4D CT Computed Ventilation to Lung Function Measured with Hyperpolarized Xenon-129 MRI

    SciTech Connect

    Neal, B; Chen, Q

    2015-06-15

    Purpose: To correlate ventilation parameters computed from 4D CT to ventilation, profusion, and gas exchange measured with hyperpolarized Xenon-129 MRI for a set of lung cancer patients. Methods: Hyperpolarized Xe-129 MRI lung scans were acquired for lung cancer patients, before and after radiation therapy, measuring ventilation, perfusion, and gas exchange. In the standard clinical workflow, these patients also received 4D CT scans before treatment. Ventilation was computed from 4D CT using deformable image registration (DIR). All phases of the 4D CT scan were registered using a B-spline deformable registration. Ventilation at the voxel level was then computed for each phase based on a Jacobian volume expansion metric, yielding phase sorted ventilation images. Ventilation based upon 4D CT and Xe-129 MRI were co-registered, allowing qualitative visual comparison and qualitative comparison via the Pearson correlation coefficient. Results: Analysis shows a weak correlation between hyperpolarized Xe-129 MRI and 4D CT DIR ventilation, with a Pearson correlation coefficient of 0.17 to 0.22. Further work will refine the DIR parameters to optimize the correlation. The weak correlation could be due to the limitations of 4D CT, registration algorithms, or the Xe-129 MRI imaging. Continued development will refine parameters to optimize correlation. Conclusion: Current analysis yields a minimal correlation between 4D CT DIR and Xe-129 MRI ventilation. Funding provided by the 2014 George Amorino Pilot Grant in Radiation Oncology at the University of Virginia.

  5. Sampling Hyperpolarized Molecules Utilizing a 1 Tesla Permanent Magnetic Field

    PubMed Central

    Tee, Sui Seng; DiGialleonardo, Valentina; Eskandari, Roozbeh; Jeong, Sangmoo; Granlund, Kristin L.; Miloushev, Vesselin; Poot, Alex J.; Truong, Steven; Alvarez, Julio A.; Aldeborgh, Hannah N.; Keshari, Kayvan R.

    2016-01-01

    Hyperpolarized magnetic resonance spectroscopy (HP MRS) using dynamic nuclear polarization (DNP) is a technique that has greatly enhanced the sensitivity of detecting 13C nuclei. However, the HP MRS polarization decays in the liquid state according to the spin-lattice relaxation time (T1) of the nucleus. Sampling of the signal also destroys polarization, resulting in a limited temporal ability to observe biologically interesting reactions. In this study, we demonstrate that sampling hyperpolarized signals using a permanent magnet at 1 Tesla (1T) is a simple and cost-effective method to increase T1s without sacrificing signal-to-noise. Biologically-relevant information may be obtained with a permanent magnet using enzyme solutions and in whole cells. Of significance, our findings indicate that changes in pyruvate metabolism can also be quantified in a xenograft model at this field strength. PMID:27597137

  6. Hyperpolarized Magnetic Resonance as a Sensitive Detector of Metabolic Function

    PubMed Central

    2015-01-01

    Hyperpolarized magnetic resonance allows for noninvasive measurements of biochemical reactions in vivo. Although this technique provides a unique tool for assaying enzymatic activities in intact organs, the scope of its application is still elusive for the wider scientific community. The purpose of this review is to provide key principles and parameters to guide the researcher interested in adopting this technology to address a biochemical, biomedical, or medical issue. It is presented in the form of a compendium containing the underlying essential physical concepts as well as suggestions to help assess the potential of the technique within the framework of specific research environments. Explicit examples are used to illustrate the power as well as the limitations of hyperpolarized magnetic resonance. PMID:25369537

  7. Sampling Hyperpolarized Molecules Utilizing a 1 Tesla Permanent Magnetic Field

    NASA Astrophysics Data System (ADS)

    Tee, Sui Seng; Digialleonardo, Valentina; Eskandari, Roozbeh; Jeong, Sangmoo; Granlund, Kristin L.; Miloushev, Vesselin; Poot, Alex J.; Truong, Steven; Alvarez, Julio A.; Aldeborgh, Hannah N.; Keshari, Kayvan R.

    2016-09-01

    Hyperpolarized magnetic resonance spectroscopy (HP MRS) using dynamic nuclear polarization (DNP) is a technique that has greatly enhanced the sensitivity of detecting 13C nuclei. However, the HP MRS polarization decays in the liquid state according to the spin-lattice relaxation time (T1) of the nucleus. Sampling of the signal also destroys polarization, resulting in a limited temporal ability to observe biologically interesting reactions. In this study, we demonstrate that sampling hyperpolarized signals using a permanent magnet at 1 Tesla (1T) is a simple and cost-effective method to increase T1s without sacrificing signal-to-noise. Biologically-relevant information may be obtained with a permanent magnet using enzyme solutions and in whole cells. Of significance, our findings indicate that changes in pyruvate metabolism can also be quantified in a xenograft model at this field strength.

  8. Optical hyperpolarization of 13C nuclear spins in nanodiamond ensembles

    NASA Astrophysics Data System (ADS)

    Chen, Q.; Schwarz, I.; Jelezko, F.; Retzker, A.; Plenio, M. B.

    2015-11-01

    Dynamical nuclear polarization holds the key for orders of magnitude enhancements of nuclear magnetic resonance signals which, in turn, would enable a wide range of novel applications in biomedical sciences. However, current implementations of DNP require cryogenic temperatures and long times for achieving high polarization. Here we propose and analyze in detail protocols that can achieve rapid hyperpolarization of 13C nuclear spins in randomly oriented ensembles of nanodiamonds at room temperature. Our protocols exploit a combination of optical polarization of electron spins in nitrogen-vacancy centers and the transfer of this polarization to 13C nuclei by means of microwave control to overcome the severe challenges that are posed by the random orientation of the nanodiamonds and their nitrogen-vacancy centers. Specifically, these random orientations result in exceedingly large energy variations of the electron spin levels that render the polarization and coherent control of the nitrogen-vacancy center electron spins as well as the control of their coherent interaction with the surrounding 13C nuclear spins highly inefficient. We address these challenges by a combination of an off-resonant microwave double resonance scheme in conjunction with a realization of the integrated solid effect which, together with adiabatic rotations of external magnetic fields or rotations of nanodiamonds, leads to a protocol that achieves high levels of hyperpolarization of the entire nuclear-spin bath in a randomly oriented ensemble of nanodiamonds even at room temperature. This hyperpolarization together with the long nuclear-spin polarization lifetimes in nanodiamonds and the relatively high density of 13C nuclei has the potential to result in a major signal enhancement in 13C nuclear magnetic resonance imaging and suggests functionalized and hyperpolarized nanodiamonds as a unique probe for molecular imaging both in vitro and in vivo.

  9. The Spin-Lattice Relaxation of Hyperpolarized 89Y Complexes

    NASA Astrophysics Data System (ADS)

    Jindal, Ashish; Lumata, Lloyd; Xing, Yixun; Merritt, Matthew; Zhao, Piyu; Malloy, Craig; Sherry, Dean; Kovacs, Zoltan

    2011-03-01

    The low sensitivity of NMR can be overcome by dynamic nuclear polarization (DNP). However, a limitation to the use of hyperpolarized materials is the signal decay due to T1 relaxation. Among NMR-active nuclei, 89 Y is potentially valuable in medical imaging because in chelated form, pH-sensitive agents can be developed. 89 Y also offers many attractive features -- 100 % abundance, a 1/2 spin, and a long T1 , up to 10 min. Yet, developing new 89 Y complexes with even longer T1 values is desirable. Designing such complexes relies upon understanding the mechanism(s) responsible for T1 relaxation. We report an approach to hyperpolarized T1 measurements that enabled an analysis of relaxation mechanisms by selective deuteration of the ligand backbone, the solvent or both. Hyperpolarized 89 Y -- DTPA, DOTA, EDTA, and deuterated EDTA complexes were studied. Results suggest that substitution of low-gamma nuclei on the ligand backbone as opposed to that of the solvent most effectively increase the 89 Y T1 . These results are encouraging for in vivo applications as the presence of bound water may not dramatically affect the T1 .

  10. Improving the Hyperpolarization of 31P Nuclei by Synthetic Design

    PubMed Central

    2015-01-01

    Traditional 31P NMR or MRI measurements suffer from low sensitivity relative to 1H detection and consequently require longer scan times. We show here that hyperpolarization of 31P nuclei through reversible interactions with parahydrogen can deliver substantial signal enhancements in a range of regioisomeric phosphonate esters containing a heteroaromatic motif which were synthesized in order to identify the optimum molecular scaffold for polarization transfer. A 3588-fold 31P signal enhancement (2.34% polarization) was returned for a partially deuterated pyridyl substituted phosphonate ester. This hyperpolarization level is sufficient to allow single scan 31P MR images of a phantom to be recorded at a 9.4 T observation field in seconds that have signal-to-noise ratios of up to 94.4 when the analyte concentration is 10 mM. In contrast, a 12 h 2048 scan measurement under standard conditions yields a signal-to-noise ratio of just 11.4. 31P-hyperpolarized images are also reported from a 7 T preclinical scanner. PMID:25811635

  11. Extending the Lifetime of Hyperpolarized Propane Gas through Reversible Dissolution.

    PubMed

    Burueva, Dudari B; Romanov, Alexey S; Salnikov, Oleg G; Zhivonitko, Vladimir V; Chen, Yu-Wen; Barskiy, Danila A; Chekmenev, Eduard Y; Hwang, Dennis W; Kovtunov, Kirill V; Koptyug, Igor V

    2017-03-02

    Hyperpolarized (HP) propane produced by the parahydrogen-induced polarization (PHIP) technique has been recently introduced as a promising contrast agent for functional lung magnetic resonance (MR) imaging. However, its short lifetime due to a spin-lattice relaxation time T1 of less than 1 s in the gas phase is a significant translational challenge for its potential biomedical applications. The previously demonstrated approach for extending the lifetime of the HP propane state through long-lived spin states allows the HP propane lifetime to be increased by a factor of ∼3. Here, we demonstrate that a remarkable increase in the propane hyperpolarization decay time at high magnetic field (7.1 T) can be achieved by its dissolution in deuterated organic solvents (acetone-d6 or methanol-d4). The approximate values of the HP decay time for propane dissolved in acetone-d6 are 35.1 and 28.6 s for the CH2 group and the CH3 group, respectively (similar values were obtained for propane dissolved in methanol-d4), which are ∼50 times larger than the gaseous propane T1 value. Furthermore, we show that it is possible to retrieve HP propane from solution to the gas phase with the preservation of hyperpolarization.

  12. Time resolved spectroscopic NMR imaging using hyperpolarized 129Xe

    NASA Astrophysics Data System (ADS)

    Han, S.; Kühn, H.; Häsing, F. W.; Münnemann, K.; Blümich, B.; Appelt, S.

    2004-04-01

    We have visualized the melting and dissolution processes of xenon (Xe) ice into different solvents using the methods of nuclear magnetic resonance (NMR) spectroscopy, imaging, and time resolved spectroscopic imaging by means of hyperpolarized 129Xe. Starting from the initial condition of a hyperpolarized solid Xe layer frozen on top of an ethanol (ethanol/water) ice block we measured the Xe phase transitions as a function of time and temperature. In the pure ethanol sample, pieces of Xe ice first fall through the viscous ethanol to the bottom of the sample tube and then form a thin layer of liquid Xe/ethanol. The xenon atoms are trapped in this liquid layer up to room temperature and keep their magnetization over a time period of 11 min. In the ethanol/water mixture (80 vol%/20%), most of the polarized Xe liquid first stays on top of the ethanol/water ice block and then starts to penetrate into the pores and cracks of the ethanol/water ice block. In the final stage, nearly all the Xe polarization is in the gas phase above the liquid and trapped inside the pores. NMR spectra of homogeneous samples of pure ethanol containing thermally polarized Xe and the spectroscopic images of the melting process show that very high concentrations of hyperpolarized Xe (about half of the density of liquid Xe) can be stored or delivered in pure ethanol.

  13. Production of nitrogen-free, hyperpolarized 129Xe gas

    NASA Astrophysics Data System (ADS)

    Ruth, U.; Hof, T.; Schmidt, J.; Fick, D.; Jänsch, H. J.

    129Xe with a nuclear polarization far above the thermal equilibrium value (hyperpolarized) is used in NMR studies to increase sensitivity. Gaseous, adsorbed, or dissolved xenon is utilized in physical, chemical, and medical applications. With the aim in mind to study single-crystal surfaces by NMR of adsorbed hyperpolarized 129Xe, three problems have to be solved. The reliable production of 129Xe with highest nuclear polarization possible, the separation of the xenon gas from the necessary quench gas nitrogen without polarization loss, and the dosing/delivery of small amounts of polarized xenon gas to a sample surface. Here we describe an optical pumping setup that regularly produces xenon gas with a 129Xe nuclear polarization of 0.7(+/-0.07). We show that a freeze-pump-thaw separation of xenon and nitrogen is feasible without a significant loss in xenon polarization. The nitrogen partial pressure can be suppressed by a factor of 400 in a single separation cycle. Dosing is achieved by using the low vapor pressure of a frozen hyperpolarized xenon sample.

  14. Constant-variable flip angles for hyperpolarized media MRI

    NASA Astrophysics Data System (ADS)

    Deng, He; Zhong, Jianping; Ruan, Weiwei; Chen, Xian; Sun, Xianping; Ye, Chaohui; Liu, Maili; Zhou, Xin

    2016-02-01

    The longitudinal magnetization of hyperpolarized media, such as hyperpolarized 129Xe, 3He, etc., is nonrenewable. When the MRI data acquisition begins at the k-domain center, a constant flip angle (CFA) results in an image of high signal-to-noise ratio (SNR) but sacrifices the accuracy of spatial information. On the other hand, a variable flip angle (VFA) strategy results in high accuracy but suffers from a low SNR. In this paper, we propose a novel scheme to optimize both the SNR and accuracy, called constant-variable flip angles (CVFA). The proposed scheme suggests that hyperpolarized magnetic resonance signals are firstly acquired through a train of n∗ CFA excitation pulses, followed by a train of N-n∗ VFA excitation pulses. We simulate and optimize the flip angle used in the CFA section, the number of CFA excitation pulses, the number of VFA excitation pulses, and the initial and final variable flip angles adopted in the VFA section. Phantom and in vivo experiments demonstrate the good performance of the CVFA designs and their ability to maintain both high SNR and spatial resolution.

  15. A hyperpolarized equilibrium for magnetic resonance.

    PubMed

    Hövener, Jan-Bernd; Schwaderlapp, Niels; Lickert, Thomas; Duckett, Simon B; Mewis, Ryan E; Highton, Louise A R; Kenny, Stephen M; Green, Gary G R; Leibfritz, Dieter; Korvink, Jan G; Hennig, Jürgen; von Elverfeldt, Dominik

    2013-01-01

    Nuclear magnetic resonance spectroscopy and imaging (MRI) play an indispensable role in science and healthcare but use only a tiny fraction of their potential. No more than ≈10 p.p.m. of all ¹H nuclei are effectively detected in a 3-Tesla clinical MRI system. Thus, a vast array of new applications lays dormant, awaiting improved sensitivity. Here we demonstrate the continuous polarization of small molecules in solution to a level that cannot be achieved in a viable magnet. The magnetization does not decay and is effectively reinitialized within seconds after being measured. This effect depends on the long-lived, entangled spin-order of parahydrogen and an exchange reaction in a low magnetic field of 10⁻³ Tesla. We demonstrate the potential of this method by fast MRI and envision the catalysis of new applications such as cancer screening or indeed low-field MRI for routine use and remote application.

  16. Noradrenaline hyperpolarizes identified rat mesopontine cholinergic neurons in vitro.

    PubMed

    Williams, J A; Reiner, P B

    1993-09-01

    Inhibition of brainstem cholinergic neurons by noradrenergic neurons of the locus ceruleus has long been suggested as a key mechanism of behavioral state control. In particular, the commonly held view is that noradrenaline (NA) plays a permissive role in rapid eye movement (REM) sleep generation by disinhibiting brainstem cholinergic neurons. While this notion has been supported by numerous investigations, the inhibition of cholinergic neurons by NA has never been directly demonstrated. The purpose of this study was to investigate the effects of NA upon identified cholinergic neurons in the rat mesopontine tegmentum. Using whole-cell patch-clamp recordings in slices, 175 cells were studied during bath application of 50 microM NA. Cholinergic neurons were positively identified by intracellular labeling with biocytin and subsequent staining with NADPH-diaphorase, a reliable marker for brainstem cholinergic neurons (Vincent et al., 1983). Successful intracellular labeling was obtained in 96 cells. Ninety-two percent (36 of 39) of cholinergic neurons hyperpolarized in response to NA, while noncholinergic cells (n = 57) exhibited mixed responses. Application of NA in a low-Ca2+, high-Mg2+ solution elicited the same hyperpolarizing effect as in normal solution, which indicated that the effect of NA on cholinergic neurons was direct. The noradrenergic hyperpolarization was mimicked by the alpha 2-adrenoceptor agonist UK-14,304, and was blocked by the alpha 2-adrenoceptor antagonist idazoxan, which suggested an alpha 2-mediated response. Finally, voltage-clamp experiments revealed that NA activates the inwardly rectifying potassium current, IKG.(ABSTRACT TRUNCATED AT 250 WORDS)

  17. Astrocytic mitochondrial membrane hyperpolarization following extended oxygen and glucose deprivation.

    PubMed

    Korenić, Andrej; Boltze, Johannes; Deten, Alexander; Peters, Myriam; Andjus, Pavle; Radenović, Lidija

    2014-01-01

    Astrocytes can tolerate longer periods of oxygen and glucose deprivation (OGD) as compared to neurons. The reasons for this reduced vulnerability are not well understood. Particularly, changes in mitochondrial membrane potential (Δψ(m)) in astrocytes, an indicator of the cellular redox state, have not been investigated during reperfusion after extended OGD exposure. Here, we subjected primary mouse astrocytes to glucose deprivation (GD), OGD and combinations of both conditions varying in duration and sequence. Changes in Δψ(m), visualized by change in the fluorescence of JC-1, were investigated within one hour after reconstitution of oxygen and glucose supply, intended to model in vivo reperfusion. In all experiments, astrocytes showed resilience to extended periods of OGD, which had little effect on Δψ(m) during reperfusion, whereas GD caused a robust Δψ(m) negativation. In case no Δψ(m) negativation was observed after OGD, subsequent chemical oxygen deprivation (OD) induced by sodium azide caused depolarization, which, however, was significantly delayed as compared to normoxic group. When GD preceded OD for 12 h, Δψ(m) hyperpolarization was induced by both GD and subsequent OD, but significant interaction between these conditions was not detected. However, when GD was extended to 48 h preceding OGD, hyperpolarization enhanced during reperfusion. This implicates synergistic effects of both conditions in that sequence. These findings provide novel information regarding the role of the two main substrates of electron transport chain (glucose and oxygen) and their hyperpolarizing effect on Δψ(m) during substrate deprivation, thus shedding new light on mechanisms of astrocyte resilience to prolonged ischemic injury.

  18. Hyperpolarized 13C Metabolic MRI of the Human Heart

    PubMed Central

    Lau, Justin Y.C.; Chen, Albert P.; Geraghty, Benjamin J.; Perks, William J.; Roifman, Idan; Wright, Graham A.; Connelly, Kim A.

    2016-01-01

    Rationale: Altered cardiac energetics is known to play an important role in the progression toward heart failure. A noninvasive method for imaging metabolic markers that could be used in longitudinal studies would be useful for understanding therapeutic approaches that target metabolism. Objective: To demonstrate the first hyperpolarized 13C metabolic magnetic resonance imaging of the human heart. Methods and Results: Four healthy subjects underwent conventional proton cardiac magnetic resonance imaging followed by 13C imaging and spectroscopic acquisition immediately after intravenous administration of a 0.1 mmol/kg dose of hyperpolarized [1-13C]pyruvate. All subjects tolerated the procedure well with no adverse effects reported ≤1 month post procedure. The [1-13C]pyruvate signal appeared within the chambers but not within the muscle. Imaging of the downstream metabolites showed 13C-bicarbonate signal mainly confined to the left ventricular myocardium, whereas the [1-13C]lactate signal appeared both within the chambers and in the myocardium. The mean 13C image signal:noise ratio was 115 for [1-13C]pyruvate, 56 for 13C-bicarbonate, and 53 for [1-13C]lactate. Conclusions: These results represent the first 13C images of the human heart. The appearance of 13C-bicarbonate signal after administration of hyperpolarized [1-13C]pyruvate was readily detected in this healthy cohort (n=4). This shows that assessment of pyruvate metabolism in vivo in humans is feasible using current technology. Clinical Trial Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT02648009. PMID:27635086

  19. The Efficiency Limits of Spin Exchange Optical Pumping Methods of 129Xe Hyperpolarization: Implications for in vivo MRI Applications

    NASA Astrophysics Data System (ADS)

    Freeman, Matthew S.

    Since the inception of hyperpolarized 129Xe MRI, the field has yearned for more efficient production of more highly polarized 129Xe. For nearly all polarizers built to date, both peak 129Xe polarization and production rate fall far below theoretical predictions. This thesis sought to develop a fundamental understanding of why the observed performance of large-scale 129Xe hyperpolarization lagged so badly behind theoretical predictions. This is done by thoroughly characterizing a high-volume, continuous-flow polarizer using optical cells having three different internal volumes, and employing two different laser sources. For each of these 6 combinations, 129Xe polarization was carefully measured as a function of production rate across a range of laser absorption levels. The resultant peak polarizations were consistently a factor of 2-3 lower than predicted across a range of absorption levels, and scaling of production rates deviated badly from predictions based on spin exchange efficiency. To bridge this gap, we propose that paramagnetic, activated Rb clusters form during spin exchange optical pumping (SEOP), and depolarize Rb and 129Xe, while unproductively scattering optical pumping light. When a model was built that incorporated the effects of clusters, its predictions matched observations for both polarization and production rate for all 6 systems studied. This permits us to place a limit on cluster number density of <2 x 109 cm-3. The work culminates with deploying this framework to identify methods to improve polarization to above 50%, leaving the SEOP cell. Combined with additional methods of preserving polarization, the polarization of a 300-mL batch of 129Xe increased from an average of 9%, before this work began, to a recent value of 34%. We anticipate that these developments will lay the groundwork for continued advancement and scaling up of SEOP-based hyperpolarization methods that may one day permit real-time, on-demand 129Xe MRI to become a reality.

  20. Inhibition of the production of endothelium-derived hyperpolarizing factor by cannabinoid receptor agonists

    PubMed Central

    Fleming, I; Schermer, B; Popp, R; Busse, R

    1999-01-01

    The endogenous cannabinoid, anandamide, has been reported to induce an 'endothelium-derived hyperpolarizing factor (EDHF)-like' relaxation in vitro. We therefore investigated the effects of cannabinoid CB1 receptor agonists; HU 210, Δ9-tetrahydrocannabinol (Δ9-THC) and anandamide, and a CB1 antagonist/inverse agonist, SR 141716A, on nitric oxide (NO) and EDHF-mediated relaxation in precontracted rings of porcine coronary, rabbit carotid and mesenteric arteries. In rings of mesenteric artery HU 210 and Δ9-THC induced endothelium- and cyclo-oxygenase-independent relaxations which were sensitive to SR 141716A. Anandamide (0.03–30 μM) induced a slowly developing, endothelium-independent relaxation which was abolished by diclofenac and was therefore mediated by cyclo-oxygenase product(s). None of the CB1 agonists tested affected the tone of precontracted rings of rabbit carotid or porcine coronary artery. In endothelium-intact segments, HU 210, Δ9-THC and anandamide did not affect NO-mediated responses but under conditions of continuous NO synthase/cyclo-oxygenase blockade, significantly inhibited acetylcholine and bradykinin-induced relaxations which are attributed to the production of EDHF. The effects of HU 210 and Δ9-THC were not observed when experiments were performed in the presence of SR 141716A suggesting the involvement of the CB1 receptor. In a patch clamp bioassay of EDHF production, HU 210 decreased the EDHF-mediated hyperpolarization of detector smooth muscle cells when applied to the donor segment but was without effect on the membrane potential of detector cells. The inhibition of EDHF production was unrelated to alterations in Ca2+-signalling or cytochrome P450 activity. These results suggest that the activation of endothelial CB1 receptors appears to be negatively coupled to the production of EDHF. PMID:10193775

  1. Fast Padé Transform Accelerated CSI for Hyperpolarized MRS

    PubMed Central

    Hansen, Esben Szocska Søvsø; Kim, Sun; Miller, Jack J.; Geferath, Marcus; Morrell, Glen; Laustsen, Christoffer

    2016-01-01

    The fast Padé transform (FPT) is a method of spectral analysis that can be used to reconstruct nuclear magnetic resonance spectra from truncated free induction decay signals with superior robustness and spectral resolution compared with conventional Fourier analysis. The aim of this study is to show the utility of FPT in reducing of the scan time required for hyperpolarized 13C chemical shift imaging (CSI) without sacrificing the ability to resolve a full spectrum. Simulations, phantom, and in vivo hyperpolarized [1-13C] pyruvate CSI data were processed with FPT and compared with conventional analysis methods. FPT shows improved stability and spectral resolution on truncated data compared with the fast Fourier transform and shows results that are comparable to those of the model-based fitting methods, enabling a reduction in the needed acquisition time in 13C CSI experiments. Using FPT can reduce the readout length in the spectral dimension by 2–6 times in 13C CSI compared with conventional Fourier analysis without sacrificing the spectral resolution. This increased speed is crucial for 13C CSI because T1 relaxation considerably limits the available scan time. In addition, FPT can also yield direct quantification of metabolite concentration without the additional peak analysis required in conventional Fourier analysis. PMID:28018967

  2. Genetically encoded reporters for hyperpolarized xenon magnetic resonance imaging

    NASA Astrophysics Data System (ADS)

    Shapiro, Mikhail G.; Ramirez, R. Matthew; Sperling, Lindsay J.; Sun, George; Sun, Jinny; Pines, Alexander; Schaffer, David V.; Bajaj, Vikram S.

    2014-07-01

    Magnetic resonance imaging (MRI) enables high-resolution non-invasive observation of the anatomy and function of intact organisms. However, previous MRI reporters of key biological processes tied to gene expression have been limited by the inherently low molecular sensitivity of conventional 1H MRI. This limitation could be overcome through the use of hyperpolarized nuclei, such as in the noble gas xenon, but previous reporters acting on such nuclei have been synthetic. Here, we introduce the first genetically encoded reporters for hyperpolarized 129Xe MRI. These expressible reporters are based on gas vesicles (GVs), gas-binding protein nanostructures expressed by certain buoyant microorganisms. We show that GVs are capable of chemical exchange saturation transfer interactions with xenon, which enables chemically amplified GV detection at picomolar concentrations (a 100- to 10,000-fold improvement over comparable constructs for 1H MRI). We demonstrate the use of GVs as heterologously expressed indicators of gene expression and chemically targeted exogenous labels in MRI experiments performed on living cells.

  3. Switchable resonant hyperpolarization transfer to 29Si spins in natural silicon

    NASA Astrophysics Data System (ADS)

    Dluhy, Phillip; Salvail, Jeff Z.; Saeedi, Kamyar; Thewalt, Mike L. W.; Simmons, Stephanie

    2015-05-01

    Silicon nano- and microparticles containing polarized 29Si spins are promising inexpensive and biocompatible medical imaging agents, particularly for magnetic resonance imaging (MRI). Maximizing out-of-equilibrium polarization (i.e., hyperpolarization) of the 29Si nuclear spins as efficiently as possible is critical for such an application. Here we identify and exploit a frequency-matched resonant transfer process between easily hyperpolarized bulk 31P and otherwise insensitive 29Si nuclear spins in natural silicon, boosting the 29Si signal to over 200 times its thermal equilibrium signal. This technique could be used in tandem with microwave-based hyperpolarization schemes for even higher efficiencies. Lastly, this hyperpolarization buildup process does not necessarily introduce an additional source of decoherence; after hyperpolarization the resonant transfer process can be switched off to recover the ultralong lifetimes of 29Si spins for in vivo imaging.

  4. Observing and preventing rubidium runaway in a direct-infusion xenon-spin hyperpolarizer optimized for high-resolution hyper-CEST (chemical exchange saturation transfer using hyperpolarized nuclei) NMR

    SciTech Connect

    Witte, C.; Kunth, M.; Rossella, F.; Schröder, L.

    2014-02-28

    Xenon is well known to undergo host-guest interactions with proteins and synthetic molecules. As xenon can also be hyperpolarized by spin exchange optical pumping, allowing the investigation of highly dilute systems, it makes an ideal nuclear magnetic resonance probe for such host molecules. The utility of xenon as a probe can be further improved using Chemical Exchange Saturation Transfer using hyperpolarized nuclei (Hyper-CEST), but for highly accurate experiments requires a polarizer and xenon infusion system optimized for such measurements. We present the design of a hyperpolarizer and xenon infusion system specifically designed to meet the requirements of Hyper-CEST measurements. One key element of this design is preventing rubidium runaway, a chain reaction induced by laser heating that prevents efficient utilization of high photon densities. Using thermocouples positioned along the pumping cell we identify the sources of heating and conditions for rubidium runaway to occur. We then demonstrate the effectiveness of actively cooling the optical cell to prevent rubidium runaway in a compact setup. This results in a 2–3-fold higher polarization than without cooling, allowing us to achieve a polarization of 25% at continuous flow rates of 9 ml/min of {sup 129}Xe. The simplicity of this design also allows it to be retrofitted to many existing polarizers. Combined with a direction infusion system that reduces shot-to-shot noise down to 0.56% we have captured Hyper-CEST spectra in unprecedented detail, allowing us to completely resolve peaks separated by just 1.62 ppm. Due to its high polarization and excellent stability, our design allows the comparison of underlying theories of host-guest systems with experiment at low concentrations, something extremely difficult with previous polarizers.

  5. Observing and preventing rubidium runaway in a direct-infusion xenon-spin hyperpolarizer optimized for high-resolution hyper-CEST (chemical exchange saturation transfer using hyperpolarized nuclei) NMR.

    PubMed

    Witte, C; Kunth, M; Rossella, F; Schröder, L

    2014-02-28

    Xenon is well known to undergo host-guest interactions with proteins and synthetic molecules. As xenon can also be hyperpolarized by spin exchange optical pumping, allowing the investigation of highly dilute systems, it makes an ideal nuclear magnetic resonance probe for such host molecules. The utility of xenon as a probe can be further improved using Chemical Exchange Saturation Transfer using hyperpolarized nuclei (Hyper-CEST), but for highly accurate experiments requires a polarizer and xenon infusion system optimized for such measurements. We present the design of a hyperpolarizer and xenon infusion system specifically designed to meet the requirements of Hyper-CEST measurements. One key element of this design is preventing rubidium runaway, a chain reaction induced by laser heating that prevents efficient utilization of high photon densities. Using thermocouples positioned along the pumping cell we identify the sources of heating and conditions for rubidium runaway to occur. We then demonstrate the effectiveness of actively cooling the optical cell to prevent rubidium runaway in a compact setup. This results in a 2-3-fold higher polarization than without cooling, allowing us to achieve a polarization of 25% at continuous flow rates of 9 ml/min of (129)Xe. The simplicity of this design also allows it to be retrofitted to many existing polarizers. Combined with a direction infusion system that reduces shot-to-shot noise down to 0.56% we have captured Hyper-CEST spectra in unprecedented detail, allowing us to completely resolve peaks separated by just 1.62 ppm. Due to its high polarization and excellent stability, our design allows the comparison of underlying theories of host-guest systems with experiment at low concentrations, something extremely difficult with previous polarizers.

  6. Molecular hydrogen and catalytic combustion in the production of hyperpolarized 83Kr and 129Xe MRI contrast agents

    PubMed Central

    Rogers, Nicola J.; Hill-Casey, Fraser; Stupic, Karl F.; Six, Joseph S.; Lesbats, Clémentine; Rigby, Sean P.; Fraissard, Jacques; Pavlovskaya, Galina E.; Meersmann, Thomas

    2016-01-01

    Hyperpolarized (hp) 83Kr is a promising MRI contrast agent for the diagnosis of pulmonary diseases affecting the surface of the respiratory zone. However, the distinct physical properties of 83Kr that enable unique MRI contrast also complicate the production of hp 83Kr. This work presents a previously unexplored approach in the generation of hp 83Kr that can likewise be used for the production of hp 129Xe. Molecular nitrogen, typically used as buffer gas in spin-exchange optical pumping (SEOP), was replaced by molecular hydrogen without penalty for the achievable hyperpolarization. In this particular study, the highest obtained nuclear spin polarizations were P = 29% for 83Kr and P = 63% for 129Xe. The results were reproduced over many SEOP cycles despite the laser-induced on-resonance formation of rubidium hydride (RbH). Following SEOP, the H2 was reactively removed via catalytic combustion without measurable losses in hyperpolarized spin state of either 83Kr or 129Xe. Highly spin-polarized 83Kr can now be purified for the first time, to our knowledge, to provide high signal intensity for the advancement of in vivo hp 83Kr MRI. More generally, a chemical reaction appears as a viable alternative to the cryogenic separation process, the primary purification method of hp 129Xe for the past 2 1/2 decades. The inherent simplicity of the combustion process will facilitate hp 129Xe production and should allow for on-demand continuous flow of purified and highly spin-polarized 129Xe. PMID:26961001

  7. Molecular hydrogen and catalytic combustion in the production of hyperpolarized 83Kr and 129Xe MRI contrast agents.

    PubMed

    Rogers, Nicola J; Hill-Casey, Fraser; Stupic, Karl F; Six, Joseph S; Lesbats, Clémentine; Rigby, Sean P; Fraissard, Jacques; Pavlovskaya, Galina E; Meersmann, Thomas

    2016-03-22

    Hyperpolarized (hp) (83)Kr is a promising MRI contrast agent for the diagnosis of pulmonary diseases affecting the surface of the respiratory zone. However, the distinct physical properties of (83)Kr that enable unique MRI contrast also complicate the production of hp (83)Kr. This work presents a previously unexplored approach in the generation of hp (83)Kr that can likewise be used for the production of hp (129)Xe. Molecular nitrogen, typically used as buffer gas in spin-exchange optical pumping (SEOP), was replaced by molecular hydrogen without penalty for the achievable hyperpolarization. In this particular study, the highest obtained nuclear spin polarizations were P =29% for(83)Kr and P= 63% for (129)Xe. The results were reproduced over many SEOP cycles despite the laser-induced on-resonance formation of rubidium hydride (RbH). Following SEOP, the H2 was reactively removed via catalytic combustion without measurable losses in hyperpolarized spin state of either (83)Kr or (129)Xe. Highly spin-polarized (83)Kr can now be purified for the first time, to our knowledge, to provide high signal intensity for the advancement of in vivo hp (83)Kr MRI. More generally, a chemical reaction appears as a viable alternative to the cryogenic separation process, the primary purification method of hp(129)Xe for the past 2 1/2 decades. The inherent simplicity of the combustion process will facilitate hp (129)Xe production and should allow for on-demand continuous flow of purified and highly spin-polarized (129)Xe.

  8. Molecular hydrogen and catalytic combustion in the production of hyperpolarized 83Kr and 129Xe MRI contrast agents

    NASA Astrophysics Data System (ADS)

    Rogers, Nicola J.; Hill-Casey, Fraser; Stupic, Karl F.; Six, Joseph S.; Lesbats, Clémentine; Rigby, Sean P.; Fraissard, Jacques; Pavlovskaya, Galina E.; Meersmann, Thomas

    2016-03-01

    Hyperpolarized (hp) 83Kr is a promising MRI contrast agent for the diagnosis of pulmonary diseases affecting the surface of the respiratory zone. However, the distinct physical properties of 83Kr that enable unique MRI contrast also complicate the production of hp 83Kr. This work presents a previously unexplored approach in the generation of hp 83Kr that can likewise be used for the production of hp 129Xe. Molecular nitrogen, typically used as buffer gas in spin-exchange optical pumping (SEOP), was replaced by molecular hydrogen without penalty for the achievable hyperpolarization. In this particular study, the highest obtained nuclear spin polarizations were P = 29% for 83Kr and P = 63% for 129Xe. The results were reproduced over many SEOP cycles despite the laser-induced on-resonance formation of rubidium hydride (RbH). Following SEOP, the H2 was reactively removed via catalytic combustion without measurable losses in hyperpolarized spin state of either 83Kr or 129Xe. Highly spin-polarized 83Kr can now be purified for the first time, to our knowledge, to provide high signal intensity for the advancement of in vivo hp 83Kr MRI. More generally, a chemical reaction appears as a viable alternative to the cryogenic separation process, the primary purification method of hp 129Xe for the past 2 1/2 decades. The inherent simplicity of the combustion process will facilitate hp 129Xe production and should allow for on-demand continuous flow of purified and highly spin-polarized 129Xe.

  9. NITRIC OXIDE, MITOCHONDRIAL HYPERPOLARIZATION AND T-CELL ACTIVATION

    PubMed Central

    Nagy, Gyorgy; Koncz, Agnes; Fernandez, David; Perl, Andras

    2007-01-01

    T lymphocyte activation is associated with nitric oxide (NO) production that plays an essential role in multiple T cell functions. NO acts as a messenger, activating soluble guanyl cyclase and participating in the transduction signaling pathways involving cyclic GMP. NO modulates mitochondrial events that are involved in apoptosis and regulates mitochondrial membrane potential and mitochondrial biogenesis in many cell types, including lymphocytes. Mitochondrial hyperpolarization (MHP), an early and reversible event during both T lymphocyte activation and apoptosis, is regulated by NO. Here, we discuss recent evidence that NO-induced MHP represents a molecular switch in multiple T cell signaling pathways. Overproduction of NO in systemic lupus erythematosus (SLE) induces mitochondrial biogenesis and alters Ca2+ signaling. Thus, while NO plays a physiological role in lymphocyte cell signaling, its overproduction may disturb normal T cell function, contributing to the pathogenesis of autoimmunity. PMID:17462531

  10. Perspectives of hyperpolarized noble gas MRI beyond 3He

    NASA Astrophysics Data System (ADS)

    Lilburn, David M. L.; Pavlovskaya, Galina E.; Meersmann, Thomas

    2013-04-01

    Nuclear Magnetic Resonance (NMR) studies with hyperpolarized (hp) noble gases are at an exciting interface between physics, chemistry, materials science and biomedical sciences. This paper intends to provide a brief overview and outlook of magnetic resonance imaging (MRI) with hp noble gases other than hp 3He. A particular focus are the many intriguing experiments with 129Xe, some of which have already matured to useful MRI protocols, while others display high potential for future MRI applications. Quite naturally for MRI applications the major usage so far has been for biomedical research but perspectives for engineering and materials science studies are also provided. In addition, the prospects for surface sensitive contrast with hp 83Kr MRI is discussed.

  11. [Microstructure of the lung: diffusion measurement of hyperpolarized 3Helium].

    PubMed

    Morbach, Andreas E; Gast, Klaus K; Schmiedeskamp, Jörg; Herweling, Annette; Windirsch, Michael; Dahmen, Anja; Ley, Sebastian; Heussel, Claus-Peter; Heil, Werner; Kauczor, Hans-Ulrich; Schreiber, Wolfgang G

    2006-01-01

    Imaging methods to study the lung are traditionally based on x-ray or on radioactive contrast agents. Conventional magnetic resonance imaging (MRI) has only limited applications for lung imaging because of the low tissue density of protons concentration of hydrogen atoms, which are usually the basis for the imaging. The introduction of hyperpolarized noble gases as a contrast agent in MRI has opened new possibilities for lung diagnosis. The present paper describes this new technique. Diffusion-weighted MRI for assessment of the lung microstructure is presented here as an example of the new possibilities of functional imaging. Studies to determine the sensitivity of the diffusion measurement and regarding the correlation with traditionally established methods are also presented, along with results of the measurement of the reproducibility determined in a clinical pilot study on healthy volunteers and patients. Furthermore, a pilot measurement of the 3He diffusion tensor in the lung is presented.

  12. NMR/MRI with hyperpolarized gas and high Tc SQUID

    DOEpatents

    Schlenga, Klaus; de Souza, Ricardo E.; Wong-Foy, Annjoe; Clarke, John; Pines, Alexander

    2000-01-01

    A method and apparatus for the detection of nuclear magnetic resonance (NMR) signals and production of magnetic resonance imaging (MRI) from samples combines the use of hyperpolarized inert gases to enhance the NMR signals from target nuclei in a sample and a high critical temperature (Tc) superconducting quantum interference device (SQUID) to detect the NMR signals. The system operates in static magnetic fields of 3 mT or less (down to 0.1 mT), and at temperatures from liquid nitrogen (77K) to room temperature. Sample size is limited only by the size of the magnetic field coils and not by the detector. The detector is a high Tc SQUID magnetometer designed so that the SQUID detector can be very close to the sample, which can be at room temperature.

  13. Hyperpolarized cesium ions doped in a glass material.

    PubMed

    Ishikawa, Kiyoshi

    2014-10-17

    Hyperpolarized (HP) (133)Cs nuclear magnetic resonance signals were measured from borosilicate glass cell walls during optical pumping of cesium vapor at high magnetic field (9.4T). Significant signal enhancements were observed when additional heating of the cell wall was provided by intense but non-resonant laser irradiation, with integrated HP (133)Cs NMR signals and line widths varying as a function of heating laser power (and hence glass temperature). Given that virtually no Cs ions would originally be present in the glass, absorbed HP Cs atoms rarely met thermally-polarized Cs ions already at the surface; thus, spin-exchange via nuclear dipole interaction cannot be the primary mechanism for injecting spin polarization into the glass. Instead, it is concluded that the absorption and transport of HP atoms into the glass material itself is the dominant mechanism of nuclear spin injection at high temperatures-the first reported experimental demonstration of such a mechanism.

  14. Optical pumping system design for large production of hyperpolarized.

    PubMed

    Ruset, I C; Ketel, S; Hersman, F W

    2006-02-10

    We present a design for a spin-exchange optical pumping system to produce large quantities of highly polarized 129Xe. Low xenon concentrations in the flowing gas mixture allow the laser to maintain high Rb polarization. The large spin-exchange rate between Rb and 129Xe through the long-lived van der Waals molecules at low pressure, combined with a high flow rate, results in large production rates of hyperpolarized xenon. We report a maximum polarization of 64% achieved for a 0.3 l/h Xe flow rate, and maximum magnetization output of 6 l/h at 22% polarization. Our findings regarding the polarization dependence on temperature, nitrogen partial pressure, and gas mixture flow velocity are also reported.

  15. Hyperpolarization of rabbit superior cervical ganglion cells due to activity of an electrogenic sodium pump

    PubMed Central

    Lees, G.M.; Wallis, D.I.

    1974-01-01

    1 The mechanisms underlying the hyperpolarization which follows depolarization of rabbit superior cervical ganglion cells by acetylcholine, have been investigated and compared with the mechanisms responsible for the hyperpolarizations induced by orthodromic stimulation of the ganglion. 2 The amplitude of the drug-induced hyperpolarization (after-hyperpolarization) was diminished when [Na+]0 and the duration of the preceding depolarization were reduced. 3 In K+-free solutions, the amplitude of the after-hyperpolarization was often diminished and its rate of development was reduced. In 12.5 mM K+-Krebs solutions, the amplitude and rate of development of the after-hyperpolarization were increased; the potential was still present when the resting potential was at or close to EK. 4 Ouabain (10 μM) prevented or greatly diminished the after-hyperpolarization. The rates of onset and decay of the after-hyperpolarization were reduced in glucose-free solutions. 5 It is, therefore, concluded that the after-hypolarization is due to the activity of an electrogenic sodium pump. 6 The positive after-potential associated with the ganglionic action potential was increased in K+-free solutions and diminished when the resting potential approached EK, indicating that it is due to a period of increased K+ conductance. In the presence of high concentrations of hexamethonium (276 μM), the P wave was not selectively depressed by ouabain and has been shown by other workers to be due to a mechanism not involving an increased potassium conductance. It is concluded, therefore, that the positive after-potential, the P wave and the after-hyperpolarization are due to different mechanisms. PMID:4823465

  16. Mechanisms of long-lasting hyperpolarizations underlying slow sleep oscillations in cat corticothalamic networks.

    PubMed Central

    Contreras, D; Timofeev, I; Steriade, M

    1996-01-01

    1. To explore the nature of the long-lasting hyperpolarizations that characterize slow oscillations in corticothalamic circuits in vivo, intracellular recordings were obtained under ketamine-xylazine anaesthesia from cortical (Cx) cells of the cat precruciate motor cortex, thalamic reticular (RE) cells from the rostrolateral sector, and thalamocortical (TC) cells from the ventrolateral (VL) nucleus. 2. Measurements in the three cell types showed input resistance (Rin) to be highest during the long-lasting hyperpolarizations that correspond to depth-positive waves of the cortical EEG. Rin was lowest during the early phase of high-amplitude depth-negative EEG waves and increased thereafter until the next cycle of the slow oscillation. 3. Spontaneous long-lasting hyperpolarizations were compared with those evoked by dorsal thalamic stimulation. Voltage versus current (V-I) plots showed similar membrane potential (Vm) ranges and slopes for spontaneous and evoked hyperpolarizations in both Cx and RE cells. V-I plots from TC cells had similar slopes, but Vm during evoked hyperpolarizations was displaced towards more negative values. 4. Intracellular injection of constant hyperpolarizing current in Cx cells increased the amplitude of the initial part of the depolarizing plateau of the slow oscillation, but decreased the amplitude of the last part. 5. These results suggest disfacilitation to be the dominant mechanism in the membrane of cortical and thalamic cells during the spontaneous long-lasting hyperpolarizations, which shape and synchronize slow oscillations in corticothalamic networks. In Cx and RE cells, the same mechanism underlies thalamically evoked long-lasting hyperpolarizations. By contrast, evoked responses in TC cells show a strong additional hyperpolarizing factor. We propose that GABAB processes are stronger in TC than in Cx neurones, thus rendering the thalamus an easier target for absence-type epileptic phenomena through potentiation of thalamic rebound

  17. Prolonged hyperpolarizing potentials precede spindle oscillations in the thalamic reticular nucleus

    PubMed Central

    Fuentealba, Pablo; Timofeev, Igor; Steriade, Mircea

    2004-01-01

    The thalamic reticular (RE) nucleus is a key structure in the generation of spindles, a hallmark bioelectrical oscillation during early stages of sleep. Intracellular recordings of RE neurons in vivo revealed the presence of prolonged hyperpolarizing potentials preceding spindles in a subgroup (30%) of neurons. These hyperpolarizations (6-10 mV) lasted for 200-300 ms and were present just before the onset of spontaneously occurring spindle waves. Corticothalamic volleys also were effective in generating such hyperpolarizations followed by spindles in RE neurons. A drop of up to 40% in the apparent input resistance (Rin) was associated with these hyperpolarizing potentials, suggesting an active process rather than disfacilitation. Accordingly, the reversal potential was approximately -100 mV for both spontaneous and cortically elicited hyperpolarizations, consistent with the activation of slow K+ conductances. QX-314 in the recording pipettes decreased both the amplitude and incidence of prolonged hyperpolarizations, suggesting the participation of G protein-dependent K+ currents in the generation of hyperpolarizations. Simultaneous extracellular and intracellular recordings in the RE nucleus demonstrated that some RE neurons discharged during the hyperpolarizations and, thus, may be implicated in their generation. The prolonged hyperpolarizations preceding spindles may play a role in the transition from tonic to bursting firing of RE neurons within a range of membrane potential (-60 to -65 mV) at which they set favorable conditions for the generation of low-threshold spike bursts that initiate spindle sequences. These data are further arguments for the generation of spindles within the thalamic RE nucleus. PMID:15210981

  18. Apparatus for preparing a solution of a hyperpolarized noble gas for NMR and MRI analysis

    DOEpatents

    Pines, Alexander; Budinger, Thomas; Navon, Gil; Song, Yi-Qiao; Appelt, Stephan; Bifone, Angelo; Taylor, Rebecca; Goodson, Boyd; Seydoux, Roberto; Room, Toomas; Pietrass, Tanja

    2008-06-10

    The present invention relates generally to nuclear magnetic resonance (NMR) techniques for both spectroscopy and imaging. More particularly, the present invention relates to methods in which hyperpolarized noble gases (e.g., Xe and He) are used to enhance and improve NMR and MRI. Additionally, the hyperpolarized gas solutions of the invention are useful both in vitro and in vivo to study the dynamics or structure of a system. When used with biological systems, either in vivo or in vitro, it is within the scope of the invention to target the hyperpolarized gas and deliver it to specific regions within the system.

  19. Enhancement of NMR and MRI in the presence of hyperpolarized noble gases

    DOEpatents

    Pines, Alexander; Budinger, Thomas; Navon, Gil; Song, Yi-Qiao; Appelt, Stephan; Bifone, Angelo; Taylor, Rebecca; Goodson, Boyd; Seydoux, Roberto; Room, Toomas; Pietrass, Tanja

    2004-11-16

    The present invention relates generally to nuclear magnetic resonance (NMR) techniques for both spectroscopy and imaging. More particularly, the present invention relates to methods in which hyperpolarized noble gases (e.g., Xe and He) are used to enhance and improve NMR and MRI. Additionally, the hyperpolarized gas solutions of the invention are useful both in vitro and in vivo to study the dynamics or structure of a system. When used with biological systems, either in vivo or in vitro, it is within the scope of the invention to target the hyperpolarized gas and deliver it to specific regions within the system.

  20. Modulation of the isoprenaline-induced membrane hyperpolarization of mouse skeletal muscle cells.

    PubMed Central

    van Mil, H. G.; Kerkhof, C. J.; Siegenbeek van Heukelom, J.

    1995-01-01

    1. The hyperpolarization of the resting membrane potential, Vm, induced by isoprenaline in the lumbrical muscle fibres of the mouse, was investigated by use of intracellular microelectrodes. 2. In normal Krebs-Henseleit solution (potassium concentration: K+o = 5.7 mM, 'control'), Vm was -7.40 +/- 0.2 mV; lowering K+o to 0.76 mM ('low K+o') resulted in either a hyperpolarization (Vm = -95.7 +/- 2.9 mV), or a depolarization (Vm = -52.0 +/- 0.3 mV). 3. Isoprenaline (> or = 200 nM) induced a hyperpolarization of Vm by delta Vm = -5.6 +/- 0.4 mV in control solution. 4. When Vm hyperpolarized after switching to low K+o, the addition of isoprenaline resulted in increased hyperpolarization Vm: delta Vm = -16.3 +/- 3.2 mV to a final Vm = -110.1 +/- 3.4 mV. Adding iso-prenaline when Vm depolarized in low K+o, leads to a hyperpolarization of either by -11.6 +/- 0.5 mV to -63.6 +/- 0.8 mV or by -51.7 +/- 2.7 mV to -106.9 +/- 3.9 mV. 5. Ouabain (0.1 to 1 mM) did not suppress the hyperpolarization by isoprenaline in 5.7 mM K+o (delta Vm = -6.7 +/- 0.4 mV) or the hyperpolarization of the depolarized cells in low K+- (delta Vm = -9.7 +/- 1.5 mV). 6. The hyperpolarization is a logarithmically decreasing function of K+o in the range between 2 and 20 mM (12 mV/decade). 7.IBMX and 8Br-cyclic AMP mimicked the response to isoprenaline whereas forskolin (FSK) induced in low K+o a hyperpolarization of -7.0 +/- 0.7 mV that could be augmented by addition of isoprenaline (delta Vm = -8.2 +/- 1.8 mV). 8. In control and low K+o, Ba2+ (0.6 mM) inhibited the hyperpolarization induced by isoprenaline, IBMX or 8Br-cyclic AMP. Other blockers of the potassium conductance such as TEA (5 mM) and apamin (0.4 microM) had no effect. 9. We conclude that in the lumbrical muscle of the mouse the isoprenaline-induced hyperpolarization is primarily due to an increase in potassium permeability. PMID:8680720

  1. Screening and Monitoring Response to Treatment Using Subsecond Molecular Imaging and Hyperpolarized Contrast Agents

    DTIC Science & Technology

    2013-05-01

    3 Table of Contents Page INTRODUCTION ...35 4 Introduction We are developing hyperpolarized MRI allowing for ultrafast (potentially sub...Vanderbilt University, Nashville, TN 37232 United States Introduction : One of the main bottlenecks for preclinical and clinical application of

  2. Separation of extra- and intracellular metabolites using hyperpolarized 13C diffusion weighted MR

    NASA Astrophysics Data System (ADS)

    Koelsch, Bertram L.; Sriram, Renuka; Keshari, Kayvan R.; Leon Swisher, Christine; Van Criekinge, Mark; Sukumar, Subramaniam; Vigneron, Daniel B.; Wang, Zhen J.; Larson, Peder E. Z.; Kurhanewicz, John

    2016-09-01

    This work demonstrates the separation of extra- and intracellular components of glycolytic metabolites with diffusion weighted hyperpolarized 13C magnetic resonance spectroscopy. Using b-values of up to 15,000 s mm-2, a multi-exponential signal response was measured for hyperpolarized [1-13C] pyruvate and lactate. By fitting the fast and slow asymptotes of these curves, their extra- and intracellular weighted diffusion coefficients were determined in cells perfused in a MR compatible bioreactor. In addition to measuring intracellular weighted diffusion, extra- and intracellular weighted hyperpolarized 13C metabolites pools are assessed in real-time, including their modulation with inhibition of monocarboxylate transporters. These studies demonstrate the ability to simultaneously assess membrane transport in addition to enzymatic activity with the use of diffusion weighted hyperpolarized 13C MR. This technique could be an indispensible tool to evaluate the impact of microenvironment on the presence, aggressiveness and metastatic potential of a variety of cancers.

  3. Design of a 15N Molecular Unit to Achieve Long Retention of Hyperpolarized Spin State

    PubMed Central

    Nonaka, Hiroshi; Hirano, Masashi; Imakura, Yuki; Takakusagi, Yoichi; Ichikawa, Kazuhiro; Sando, Shinsuke

    2017-01-01

    Nuclear hyperpolarization is a phenomenon that can be used to improve the sensitivity of magnetic resonance molecular sensors. However, such sensors typically suffer from short hyperpolarization lifetime. Herein we report that [15N, D14]trimethylphenylammonium (TMPA) has a remarkably long spin–lattice relaxation time (1128 s, 14.1 T, 30 °C, D2O) on its 15N nuclei and achieves a long retention of the hyperpolarized state. [15N, D14]TMPA-based hyperpolarized sensor for carboxylesterase allowed the highly sensitive analysis of enzymatic reaction by 15N NMR for over 40 min in phophate-buffered saline (H2O, pH 7.4, 37 °C). PMID:28067292

  4. Pulmonary MRI contrast using Surface Quadrupolar Relaxation (SQUARE) of hyperpolarized (83)Kr.

    PubMed

    Six, Joseph S; Hughes-Riley, Theodore; Lilburn, David M L; Dorkes, Alan C; Stupic, Karl F; Shaw, Dominick E; Morris, Peter G; Hall, Ian P; Pavlovskaya, Galina E; Meersmann, Thomas

    2014-01-01

    Hyperpolarized (83)Kr has previously been demonstrated to enable MRI contrast that is sensitive to the chemical composition of the surface in a porous model system. Methodological advances have lead to a substantial increase in the (83)Kr hyperpolarization and the resulting signal intensity. Using the improved methodology for spin exchange optical pumping of isotopically enriched (83)Kr, internal anatomical details of ex vivo rodent lung were resolved with hyperpolarized (83)Kr MRI after krypton inhalation. Different (83)Kr relaxation times were found between the main bronchi and the parenchymal regions in ex vivo rat lungs. The T1 weighted hyperpolarized (83)Kr MRI provided a first demonstration of surface quadrupolar relaxation (SQUARE) pulmonary MRI contrast.

  5. Design of a 15N Molecular Unit to Achieve Long Retention of Hyperpolarized Spin State

    NASA Astrophysics Data System (ADS)

    Nonaka, Hiroshi; Hirano, Masashi; Imakura, Yuki; Takakusagi, Yoichi; Ichikawa, Kazuhiro; Sando, Shinsuke

    2017-01-01

    Nuclear hyperpolarization is a phenomenon that can be used to improve the sensitivity of magnetic resonance molecular sensors. However, such sensors typically suffer from short hyperpolarization lifetime. Herein we report that [15N, D14]trimethylphenylammonium (TMPA) has a remarkably long spin–lattice relaxation time (1128 s, 14.1 T, 30 °C, D2O) on its 15N nuclei and achieves a long retention of the hyperpolarized state. [15N, D14]TMPA-based hyperpolarized sensor for carboxylesterase allowed the highly sensitive analysis of enzymatic reaction by 15N NMR for over 40 min in phophate-buffered saline (H2O, pH 7.4, 37 °C).

  6. Vascular hyperpolarization in human physiology and cardiovascular risk conditions and disease.

    PubMed

    Schinzari, F; Tesauro, M; Cardillo, C

    2017-01-01

    Hyperpolarization causing smooth muscle relaxation contributes to the maintenance of vascular homeostasis, particularly in small-calibre arteries and arterioles. It may also become a compensatory vasodilator mechanism upregulated in states with impaired nitric oxide (NO) availability. Bioassay of vascular hyperpolarization in the human circulation has been hampered by the complexity of mechanisms involved and the limited availability of investigational tools. Firm evidence, however, supports the notion that hyperpolarization participates in the regulation of resting vasodilator tone and vascular reactivity in healthy subjects. In addition, an enhanced endothelium-derived hyperpolarization contributes to both resting and agonist-stimulated vasodilation in a variety of cardiovascular risk conditions and disease. Thus, hyperpolarization mediated by epoxyeicosatrienoic acids (EETs) and H2 O2 has been observed in coronary arterioles of patients with coronary artery disease. Similarly, ouabain-sensitive and EETs-mediated hyperpolarization has been observed to compensate for NO deficiency in patients with essential hypertension. Moreover, in non-hypertensive patients with multiple cardiovascular risk factors and in hypercholesterolaemia, KCa channel-mediated vasodilation appears to be activated. A novel paradigm establishes that perivascular adipose tissue (PVAT) is an additional regulator of vascular tone/function and endothelium is not the only agent in vascular hyperpolarization. Indeed, some PVAT-derived relaxing substances, such as adiponectin and angiotensin 1-7, may exert anticontractile and vasodilator actions by the opening of KCa channels in smooth muscle cells. Conversely, PVAT-derived factors impair coronary vasodilation via differential inhibition of some K(+) channels. In view of adipose tissue abnormalities occurring in human obesity, changes in PVAT-dependent hyperpolarization may be relevant for vascular dysfunction also in this condition.

  7. Discovery of Hyperpolarized Molecular Imaging Biomarkers in a Novel Prostate Tissue Slice Culture Model

    DTIC Science & Technology

    2013-06-01

    compatible bioreactor optimized in year 1 to identify hyperpolarized metabolic biomarkers of prostate cancer presence and aggressiveness. To...accomplish this goal my group finished the engineering of a 5 mm bioreactor and acquired hyperpolarized [1-13C]pyruvate data indicating that similar signal...to noise and quality data can be achieved with 4 to 5 prostate tissue slices in the 5 mm bioreactor as was acquired from 30-40 tissue slices in the

  8. Discovery of Hyperpolarized Molecular Imaging Biomarkers in a Novel Prostate Tissue Slice Culture Model

    DTIC Science & Technology

    2013-06-01

    bioreactor optimized in year 1 to identify hyperpolarized metabolic biomarkers of prostate cancer presence and aggressiveness. To accomplish this goal my...group finished the engineering of a 5 mm bioreactor and acquired hyperpolarized [1-13C]pyruvate data indicating that similar signal to noise and...quality data can be achieved with 4 to 5 prostate tissue slices in the 5 mm bioreactor as was acquired from 30-40 tissue slices in the prior 10 mm

  9. Discovery of Hyperpolarized Molecular Imaging Biomarkers in a Novel Prostate Tissue Slice Culture Model

    DTIC Science & Technology

    2013-06-01

    compatible bioreactor and that hyperpolarized 13C spectroscopy could be employed to study real-time metabolism of normal and malignant tissues. The...function of prostate tissue slice cultures (TCSs) in an nuclear magnetic resonance (NMR)-compatible, 3-dimensional tissue culture bioreactor , (2) to use...the TSC/NMR bioreactor model to identify hyperpolarized metabolic biomarkers of prostate cancer presence and aggressiveness, and (3) to use the TSC

  10. Irreversible Catalyst Activation Enables Hyperpolarization and Water Solubility for NMR Signal Amplification by Reversible Exchange

    DTIC Science & Technology

    2016-09-12

    Irreversible Catalyst Activation Enables Hyperpolarization and Water Solubility for NMR Signal Amplification by Reversible Exchange Milton L. Truong...Supporting Information ABSTRACT: Activation of a catalyst [IrCl(COD)(IMes)] (IMes = 1,3-bis(2,4,6-trimethylphenyl)imidazol-2-ylidene; COD = cyclooctadiene...for signal amplification by reversible exchange (SABRE) was monitored by in situ hyperpolarized proton NMR at 9.4 T. During the catalyst -activation

  11. Discovery of Hyperpolarized Molecular Imaging Biomarkers in a Novel Prostate Tissue Slice Culture Model

    DTIC Science & Technology

    2011-06-01

    Ronen’s report) above the rotor and pushing the tissues out the sample tube into the rotor using a small plastic rod. Following the insertion of each...for pathology and LDH enzyme activity. Two pathologists used a five-point scale (1= excellent, 5 = poor) to quantify the quality of the pathology of...AUC), time to maximum hyperpolarized lactate, maximum hyperpolarized lactate peak, and LDH activity were statistically compared between benign and

  12. Distal airways in humans: dynamic hyperpolarized 3He MR imaging--feasibility

    NASA Technical Reports Server (NTRS)

    Tooker, Angela C.; Hong, Kwan Soo; McKinstry, Erin L.; Costello, Philip; Jolesz, Ferenc A.; Albert, Mitchell S.

    2003-01-01

    Dynamic hyperpolarized helium 3 (3He) magnetic resonance (MR) imaging of the human airways is achieved by using a fast gradient-echo pulse sequence during inhalation. The resulting dynamic images show differential contrast enhancement of both distal airways and the lung periphery, unlike static hyperpolarized 3He MR images on which only the lung periphery is seen. With this technique, up to seventh-generation airway branching can be visualized. Copyright RSNA, 2003.

  13. Flow Cytometry Analysis Reveals That Only a Subpopulation of Mouse Sperm Undergoes Hyperpolarization During Capacitation1

    PubMed Central

    Escoffier, Jessica; Navarrete, Felipe; Haddad, Doug; Santi, Celia M.; Darszon, Alberto; Visconti, Pablo E.

    2015-01-01

    To gain fertilizing capacity, mammalian sperm should reside in the female tract for a period of time. The physiological changes that render the sperm able to fertilize are known as capacitation. Capacitation is associated with an increase in intracellular pH, an increase in intracellular calcium, and phosphorylation of different proteins. This process is also accompanied by the hyperpolarization of the sperm plasma membrane potential (Em). In the present work, we used flow cytometry to analyze changes in sperm Em during capacitation in individual cells. Our results indicate that a subpopulation of hyperpolarized mouse sperm can be clearly distinguished by sperm flow cytometry analysis. Using sperm bearing green fluorescent protein in their acrosomes, we found that this hyperpolarized subpopulation is composed of sperm with intact acrosomes. In addition, we show that the capacitation-associated hyperpolarization is blocked by high extracellular K+, by PKA inhibitors, and by SLO3 inhibitors in CD1 mouse sperm, and undetectable in Slo3 knockout mouse sperm. On the other hand, in sperm incubated in conditions that do not support capacitation, sperm membrane hyperpolarization can be induced by amiloride, high extracellular NaHCO3, and cAMP agonists. Altogether, our observations are consistent with a model in which sperm Em hyperpolarization is downstream of a cAMP-dependent pathway and is mediated by the activation of SLO3 K+ channels. PMID:25855261

  14. Signal-to-noise ratio comparison of encoding methods for hyperpolarized noble gas MRI

    NASA Technical Reports Server (NTRS)

    Zhao, L.; Venkatesh, A. K.; Albert, M. S.; Panych, L. P.

    2001-01-01

    Some non-Fourier encoding methods such as wavelet and direct encoding use spatially localized bases. The spatial localization feature of these methods enables optimized encoding for improved spatial and temporal resolution during dynamically adaptive MR imaging. These spatially localized bases, however, have inherently reduced image signal-to-noise ratio compared with Fourier or Hadamad encoding for proton imaging. Hyperpolarized noble gases, on the other hand, have quite different MR properties compared to proton, primarily the nonrenewability of the signal. It could be expected, therefore, that the characteristics of image SNR with respect to encoding method will also be very different from hyperpolarized noble gas MRI compared to proton MRI. In this article, hyperpolarized noble gas image SNRs of different encoding methods are compared theoretically using a matrix description of the encoding process. It is shown that image SNR for hyperpolarized noble gas imaging is maximized for any orthonormal encoding method. Methods are then proposed for designing RF pulses to achieve normalized encoding profiles using Fourier, Hadamard, wavelet, and direct encoding methods for hyperpolarized noble gases. Theoretical results are confirmed with hyperpolarized noble gas MRI experiments. Copyright 2001 Academic Press.

  15. Real-Time MRI-Guided Catheter Tracking Using Hyperpolarized Silicon Particles

    PubMed Central

    Whiting, Nicholas; Hu, Jingzhe; Shah, Jay V.; Cassidy, Maja C.; Cressman, Erik; Zacharias Millward, Niki; Menter, David G.; Marcus, Charles M.; Bhattacharya, Pratip K.

    2015-01-01

    Visualizing the movement of angiocatheters during endovascular interventions is typically accomplished using x-ray fluoroscopy. There are many potential advantages to developing magnetic resonance imaging-based approaches that will allow three-dimensional imaging of the tissue/vasculature interface while monitoring other physiologically-relevant criteria, without exposing the patient or clinician team to ionizing radiation. Here we introduce a proof-of-concept development of a magnetic resonance imaging-guided catheter tracking method that utilizes hyperpolarized silicon particles. The increased signal of the silicon particles is generated via low-temperature, solid-state dynamic nuclear polarization, and the particles retain their enhanced signal for ≥40 minutes—allowing imaging experiments over extended time durations. The particles are affixed to the tip of standard medical-grade catheters and are used to track passage under set distal and temporal points in phantoms and live mouse models. With continued development, this method has the potential to supplement x-ray fluoroscopy and other MRI-guided catheter tracking methods as a zero-background, positive contrast agent that does not require ionizing radiation. PMID:26239953

  16. Regional fractional ventilation mapping in spontaneously breathing mice using hyperpolarized ¹²⁹Xe MRI.

    PubMed

    Imai, Hirohiko; Matsumoto, Hironobu; Miyakoshi, Erika; Okumura, Shintaro; Fujiwara, Hideaki; Kimura, Atsuomi

    2015-01-01

    The feasibility of ventilation imaging with hyperpolarized (HP) (129) Xe MRI has been investigated for quantitative and regional assessment of ventilation in spontaneously breathing mice. The multiple breath ventilation imaging technique was modified to the protocol of spontaneous inhalation of HP (129) Xe delivered continuously from a (129) Xe polarizer. A series of (129) Xe ventilation images was obtained by varying the number of breaths before the (129) Xe lung imaging. The fractional ventilation, r, was successfully evaluated for spontaneously breathing mice. An attempt was made to detect ventilation dysfunction in the emphysematous mouse lung induced by intratracheal administration of porcine pancreatic elastase (PPE). As a result, the distribution of fractional ventilation could be visualized by the r map. Significant dysfunction of ventilation was quantitatively identified in the PPE-treated group. The whole-lung r value of 0.34 ± 0.01 for control mice (N = 4) was significantly reduced, to 0.25 ± 0.07, in PPE-treated mice (N = 4) (p = 0.038). This study is the first application of multiple breath ventilation imaging to spontaneously breathing mice, and shows that this methodology is sensitive to differences in the pulmonary ventilation. This methodology is expected to improve simplicity as well as noninvasiveness when assessing regional ventilation in small rodents.

  17. Real-Time MRI-Guided Catheter Tracking Using Hyperpolarized Silicon Particles

    NASA Astrophysics Data System (ADS)

    Whiting, Nicholas; Hu, Jingzhe; Shah, Jay V.; Cassidy, Maja C.; Cressman, Erik; Zacharias Millward, Niki; Menter, David G.; Marcus, Charles M.; Bhattacharya, Pratip K.

    2015-08-01

    Visualizing the movement of angiocatheters during endovascular interventions is typically accomplished using x-ray fluoroscopy. There are many potential advantages to developing magnetic resonance imaging-based approaches that will allow three-dimensional imaging of the tissue/vasculature interface while monitoring other physiologically-relevant criteria, without exposing the patient or clinician team to ionizing radiation. Here we introduce a proof-of-concept development of a magnetic resonance imaging-guided catheter tracking method that utilizes hyperpolarized silicon particles. The increased signal of the silicon particles is generated via low-temperature, solid-state dynamic nuclear polarization, and the particles retain their enhanced signal for ≥40 minutes—allowing imaging experiments over extended time durations. The particles are affixed to the tip of standard medical-grade catheters and are used to track passage under set distal and temporal points in phantoms and live mouse models. With continued development, this method has the potential to supplement x-ray fluoroscopy and other MRI-guided catheter tracking methods as a zero-background, positive contrast agent that does not require ionizing radiation.

  18. Relaxation of hyperpolarized 129 Xe in a deflating polymer bag

    NASA Astrophysics Data System (ADS)

    Möller, Harald E.; Cleveland, Zackary I.; Driehuys, Bastiaan

    2011-09-01

    In magnetic resonance imaging with hyperpolarized (HP) noble gases, data is often acquired during prolonged gas delivery from a storage reservoir. However, little is known about the extent to which relaxation within the reservoir will limit the useful acquisition time. For quantitative characterization, 129Xe relaxation was studied in a bag made of polyvinyl fluoride (Tedlar). Particular emphasis was on wall relaxation, as this mechanism is expected to dominate. The HP 129Xe magnetization dynamics in the deflating bag were accurately described by a model assuming dissolution of Xe in the polymer matrix and dipolar relaxation with neighboring nuclear spins. In particular, the wall relaxation rate changed linearly with the surface-to-volume ratio and exhibited a relaxivity of κ = 0.392 ± 0.008 cm/h, which is in reasonable agreement with κ = 0.331 ± 0.051 cm/h measured in a static Tedlar bag. Estimates for the bulk gas-phase 129Xe relaxation yielded T1bulk=2.55±0.22 h, which is dominated by intrinsic Xe-Xe relaxation, with small additional contributions from magnetic field inhomogeneities and oxygen-induced relaxation. Calculations based on these findings indicate that relaxation may limit HP 129Xe experiments when slow gas delivery rates are employed as, for example, in mouse imaging or vascular infusion experiments.

  19. Reducing Relaxation of Hyperpolarized ^129 Xe during Cryogenic Separation

    NASA Astrophysics Data System (ADS)

    Patton, B.; Kuzma, N. N.; Happer, W.

    2004-05-01

    Recent experimental results^1 indicate that the T1 relaxation time of solid ^129Xe is much shorter than previous models had predicted^2 near the xenon melting point of 161 K and at low magnetic fields. This enhanced relaxation is detrimental to commercial xenon polarizers, which cryogenically distill hyperpolarized ^129Xe from a buffer gas mixture. We have measured the fraction of xenon polarization lost during a typical cryogenic collection, using different permanent magnets to vary the holding field from 700 gauss to over a tesla. The results indicate that using a stronger permanent magnet around the cryo-trap is a simple way to increase the final polarization of the pure xenon gas. An additional experiment was conducted in order to determine whether the majority of the xenon relaxation occurs throughout accumulation (possibly as a result of temperature inhomogeneities within the frozen sample) or during the brief thawing time. In pinpointing the polarization loss, this research may suggest new designs for xenon polarizers. 1. Kuzma et al., Phys. Rev. Lett. 88, 147602 (2002). 2. Fitzgerald et al., Phys. Rev. B 59, 8795 (1999).

  20. Systematic T1 improvement for hyperpolarized 129xenon

    NASA Astrophysics Data System (ADS)

    Repetto, Maricel; Babcock, Earl; Blümler, Peter; Heil, Werner; Karpuk, Sergei; Tullney, Kathlynne

    2015-03-01

    The spin-lattice relaxation time T1 of hyperpolarized (HP)-129Xe was improved at typical storage conditions (i.e. low and homogeneous magnetic fields). Very long wall relaxation times T1wall of about 18 h were observed in uncoated, spherical GE180 glass cells of ∅ = 10 cm which were free of rubidium and not permanently sealed but attached to a standard glass stopcock. An "aging" process of the wall relaxation was identified by repeating measurements on the same cell. This effect could be easily removed by repeating the initial cleaning procedure. In this way, a constant wall relaxation was ensured. The Xe nuclear spin-relaxation rate 1 /T1Xel -Xe due to van der Waals molecules was investigated too, by admixing three different buffer gases (N2, SF6 and CO2). Especially CO2 exhibited an unexpected high efficiency (r) in shortening the lifetime of the Xe-Xe dimers and hence prolonging the total T1 relaxation even further. These measurements also yielded an improved accuracy for the van der Waals relaxation for pure Xe (with 85% 129Xe) of T1Xe -Xe = (4.6 ± 0.1)h . Repeating the measurements with HP 129Xe in natural abundance in mixtures with SF6, a strong dependence of T1Xe -Xe and r on the isotopic enrichment was observed, uncovering a shorter T1Xe -Xe relaxation for the 129Xe in natural composition as compared to the 85% isotopically enriched gas.

  1. Hyperpolarized 3He diffusion MRI and histology in pulmonary emphysema.

    PubMed

    Woods, Jason C; Choong, Cliff K; Yablonskiy, Dmitriy A; Bentley, John; Wong, Jonathan; Pierce, John A; Cooper, Joel D; Macklem, Peter T; Conradi, Mark S; Hogg, James C

    2006-12-01

    Diffusion MRI of hyperpolarized (3)He shows that the apparent diffusion coefficient (ADC) of (3)He gas is highly restricted in the normal lung and becomes nearly unrestricted in severe emphysema. The nature of this restricted diffusion provides information about lung structure; however, no direct comparison with histology in human lungs has been reported. The purpose of this study is to provide information about (3)He gas diffusivity in explanted human lungs, and describe the relationship between (3)He diffusivity and the surface area to lung volume ratio (SA/V) and mean linear intercept (L(m)) measurements--the gold standard for diagnosis of emphysema. Explanted lungs from patients who were undergoing lung transplantation for advanced COPD, and donor lungs that were not used for transplantation were imaged via (3)He diffusion MRI. Histological measurements were made on the same specimens after they were frozen in the position of study. There is an inverse correlation between diffusivity and SA/V (and a positive correlation between diffusivity and L(m)). An important result is that restricted (3)He diffusivity separated normal from emphysematous lung tissue more clearly than the morphometric analyses. This effect may be due to the smaller histologic sampling size compared to the MRI voxel sizes.

  2. Exploring hyperpolarized 83Kr by remotely detected NMR relaxometry

    NASA Astrophysics Data System (ADS)

    Cleveland, Zackary I.; Pavlovskaya, Galina E.; Stupic, Karl F.; LeNoir, Catherine F.; Meersmann, Thomas

    2006-01-01

    For the first time, a hyperpolarized (hp) noble gas with a nuclear electric quadrupole moment is available for high-field nuclear-magnetic-resonance (NMR) spectroscopy and magnetic-resonance imaging. Hp Kr83 (I=9/2) is generated by spin-exchange optical pumping and separated from the rubidium vapor used in the pumping process. Optical pumping occurs under the previously unstudied condition of high krypton gas densities. Signal enhancements of more than three orders of magnitude compared to the thermal equilibrium Kr83 signal at 9.4T magnetic-field strength are obtained. The spin-lattice relaxation of Kr83 is caused primarly by quadrupolar couplings during the brief adsorption periods of the krypton atoms on the surrounding container walls and significantly limits the currently obtained spin polarization. Measurements in macroscopic glass containers and in desiccated canine lung tissue at field strengths between 0.05 and 3T using remotely detected hp Kr83 NMR spectroscopy reveal that the longitudinal relaxation dramatically accelerates as the magnetic-field strength decreases.

  3. Detection of tobacco smoke deposition by hyperpolarized krypton-83 MRI.

    PubMed

    Cleveland, Zackary I; Pavlovskaya, Galina E; Stupic, Karl F; Wooten, Jan B; Repine, John E; Meersmann, Thomas

    2008-02-01

    Despite the importance of the tobacco smoke particulate matter in the lungs to the etiology of pulmonary disease in cigarette smokers, little is currently known about the spatial distribution of particle deposition or the persistence of the resulting deposits in humans, and no satisfactory technique currently exists to directly observe tobacco smoke condensate in airways. In this proof-of-principle work, hyperpolarized (hp) 83Kr MRI and NMR spectroscopy are introduced as probes for tobacco smoke deposition in porous media. A reduction in the hp-83Kr longitudinal (T1) relaxation of up to 95% under near-ambient humidity, pressure and temperature conditions was observed when the krypton gas was brought into contact with surfaces that had been exposed to cigarette smoke. This smoke-induced acceleration of the 83Kr self-relaxation was observed for model glass surfaces that, in some experiments, were coated with bovine lung surfactant extract. However, a similar effect was not observed with hp-(129)Xe indicating that the 83Kr sensitivity to smoke deposition was not caused by paramagnetic species but rather by quadrupolar relaxation due to high adsorption affinity for the smoke deposits. The 83Kr T1 differences between smoke-treated and untreated surfaces were sufficient to produce a strong contrast in variable flip angle FLASH hp-83Kr MRI, suggesting that hp-83Kr may be a promising contrast agent for in vivo pulmonary MRI.

  4. Hyperpolarized 129Xe NMR signature of living biological cells.

    PubMed

    Boutin, Céline; Desvaux, Hervé; Carrière, Marie; Leteurtre, François; Jamin, Nadège; Boulard, Yves; Berthault, Patrick

    2011-12-01

    We show that the differentiation between internal and external compartments of various biological cells in suspension can be made via simple NMR spectra of hyperpolarized (129) Xe. The spectral separation between the signals of (129) Xe in these two compartments is already known for red blood cells, because of the strong interaction of the noble gas with hemoglobin. The observation of two separate peaks in the 200-ppm region can be seen with both eukaryotic and prokaryotic cells, some of which are not known to contain paramagnetic proteins in large quantities. Using different experiments in which the cells are lysed, swell or are blocked in G2 phase, we demonstrate that the low-field-shifted peak observed corresponds to xenon in the aqueous pool inside the cells and not in the membranes. The presence of this additional peak is a clear indication of cell integrity, and its integration allows the quantification of the total cell volume. The relaxation time of intracellular xenon is sufficiently long to open up promising perspectives for cell characterization. The exchange time between the inner and outer cell compartments (on the order of 30 ms) renders possible the targeting of intracellular receptors, whereas the observation of chemical shift variations represents a method of revealing the presence of toxic species in the cells.

  5. Is my lung function really that good? Flow-type spirometer problems that elevate test results.

    PubMed

    Townsend, Mary C; Hankinson, John L; Lindesmith, Larry A; Slivka, William A; Stiver, Gregg; Ayres, Gerald T

    2004-05-01

    Most spirometry errors reduce test results, and it is widely assumed that measurement accuracy is guaranteed by frequent spirometer calibrations or calibration checks. However, zero errors and changes in flow-type spirometer sensors may occur during testing that significantly elevate test results, even though the spirometer was calibrated recently. To draw attention to these often-unrecognized problems, this report presents anomalous spirograms and test results obtained from occupational medicine clinics and hospital pulmonary function laboratories during quality assurance spirogram reviews. The spurious results appear to have been caused by inaccurate zeroing of the flow sensor, or by condensation, mucus deposition, or unstable calibration of various flow-type spirometers. These errors elevated some FVCs to 144 to 204% of predicted and probably caused 40% of 121 middle-aged working men in respirator medical clearance programs to record both FVC and FEV1 > 120% of predicted. Since spirometers report the largest values from a test, these errors must be recognized and deleted to avoid false-negative interpretations. Flow-type spirometer users at all levels, from the technician to the interpreter of test results, should be aware of the potential for and the appearance of these errors in spirograms.

  6. Systematic T1 improvement for hyperpolarized 129xenon.

    PubMed

    Repetto, Maricel; Babcock, Earl; Blümler, Peter; Heil, Werner; Karpuk, Sergei; Tullney, Kathlynne

    2015-03-01

    The spin-lattice relaxation time T1 of hyperpolarized (HP)-(129)Xe was improved at typical storage conditions (i.e. low and homogeneous magnetic fields). Very long wall relaxation times T(1)(wall) of about 18 h were observed in uncoated, spherical GE180 glass cells of ∅=10 cm which were free of rubidium and not permanently sealed but attached to a standard glass stopcock. An "aging" process of the wall relaxation was identified by repeating measurements on the same cell. This effect could be easily removed by repeating the initial cleaning procedure. In this way, a constant wall relaxation was ensured. The Xe nuclear spin-relaxation rate 1/T1(Xe-Xe) due to van der Waals molecules was investigated too, by admixing three different buffer gases (N(2), SF(6) and CO(2)). Especially CO(2) exhibited an unexpected high efficiency (r) in shortening the lifetime of the Xe-Xe dimers and hence prolonging the total T1 relaxation even further. These measurements also yielded an improved accuracy for the van der Waals relaxation for pure Xe (with 85% (129)Xe) of T(1)(Xe-Xe)=(4.6±0.1)h. Repeating the measurements with HP (129)Xe in natural abundance in mixtures with SF6, a strong dependence of T(1)(Xe-Xe) and r on the isotopic enrichment was observed, uncovering a shorter T(1)(Xe-Xe) relaxation for the (129)Xe in natural composition as compared to the 85% isotopically enriched gas.

  7. Oxygen-dependent hyperpolarized (129) Xe brain MR.

    PubMed

    Li, Haidong; Zhang, Zhiying; Zhong, Jianping; Ruan, Weiwei; Han, Yeqing; Sun, Xianping; Ye, Chaohui; Zhou, Xin

    2016-03-01

    Hyperpolarized (HP) (129) Xe MR offers unique advantages for brain functional imaging (fMRI) because of its extremely high sensitivity to different chemical environments and the total absence of background noise in biological tissues. However, its advancement and applications are currently plagued by issues of signal strength. Generally, xenon atoms found in the brain after inhalation are transferred from the lung via the bloodstream. The longitudinal relaxation time (T1 ) of HP (129) Xe is inversely proportional to the pulmonary oxygen concentration in the lung because oxygen molecules are paramagnetic. However, the T1 of (129) Xe is proportional to the pulmonary oxygen concentration in the blood, because the higher pulmonary oxygen concentration will result in a higher concentration of diamagnetic oxyhemoglobin. Accordingly, there should be an optimal pulmonary oxygen concentration for a given quantity of HP (129) Xe in the brain. In this study, the relationship between pulmonary oxygen concentration and HP (129) Xe signal in the brain was analyzed using a theoretical model and measured through in vivo experiments. The results from the theoretical model and experiments in rats are found to be in good agreement with each other. The optimal pulmonary oxygen concentration predicted by the theoretical model was 21%, and the in vivo experiments confirmed the presence of such an optimal ratio by reporting measurements between 25% and 35%. These findings are helpful for improving the (129) Xe signal in the brain and make the most of the limited spin polarization available for brain experiments. Copyright © 2016 John Wiley & Sons, Ltd.

  8. Modulation of voltage-gated sodium channels hyperpolarizes the voltage threshold for activation in spinal motoneurones.

    PubMed

    Power, Kevin E; Carlin, Kevin P; Fedirchuk, Brent

    2012-03-01

    Previous work has shown that motoneurone excitability is enhanced by a hyperpolarization of the membrane potential at which an action potential is initiated (V(th)) at the onset, and throughout brainstem-evoked fictive locomotion in the adult decerebrate cat and neonatal rat. Modeling work has suggested the modulation of Na(+) conductance as a putative mechanism underlying this state-dependent change in excitability. This study sought to determine whether modulation of voltage-gated sodium channels could induce V(th) hyperpolarization. Whole-cell patch-clamp recordings were made from antidromically identified lumbar spinal motoneurones in an isolated neonatal rat spinal cord preparation. Recordings were made with and without the bath application of veratridine, a plant alkaloid neurotoxin that acts as a sodium channel modulator. As seen in HEK 293 cells expressing Nav1.2 channels, veratridine-modified channels demonstrated a hyperpolarizing shift in their voltage-dependence of activation and a slowing of inactivation that resulted in an enhanced inward current in response to voltage ramp stimulations. In the native rat motoneurones, veratridine-modified sodium channels induced a hyperpolarization of V(th) in all 29 neonatal rat motoneurones examined (mean hyperpolarization: -6.6 ± 4.3 mV). V(th) hyperpolarization was not due to the effects on Ca(2+) and/or K(+) channels as blockade of these currents did not alter V(th). Veratridine also significantly increased the amplitude of persistent inward currents (PICs; mean increase: 72.5 ± 98.5 pA) evoked in response to slow depolarizing current ramps. However, the enhancement of the PIC amplitude had a slower time course than the hyperpolarization of V(th), and the PIC onset voltage could be either depolarized or hyperpolarized, suggesting that PIC facilitation did not mediate the V(th) hyperpolarization. We therefore suggest that central neuronal circuitry in mammals could affect V(th) in a mechanism similar to that of

  9. High-resolution low-field molecular magnetic resonance imaging of hyperpolarized liquids.

    PubMed

    Coffey, Aaron M; Kovtunov, Kirill V; Barskiy, Danila A; Koptyug, Igor V; Shchepin, Roman V; Waddell, Kevin W; He, Ping; Groome, Kirsten A; Best, Quinn A; Shi, Fan; Goodson, Boyd M; Chekmenev, Eduard Y

    2014-09-16

    We demonstrate the feasibility of microscale molecular imaging using hyperpolarized proton and carbon-13 MRI contrast media and low-field (47.5 mT) preclinical scale (38 mm i.d.) 2D magnetic resonance imaging (MRI). Hyperpolarized proton images with 94 × 94 μm(2) spatial resolution and hyperpolarized carbon-13 images with 250 × 250 μm(2) in-plane spatial resolution were recorded in 4-8 s (largely limited by the electronics response), surpassing the in-plane spatial resolution (i.e., pixel size) achievable with micro-positron emission tomography (PET). These hyperpolarized proton and (13)C images were recorded using large imaging matrices of up to 256 × 256 pixels and relatively large fields of view of up to 6.4 × 6.4 cm(2). (13)C images were recorded using hyperpolarized 1-(13)C-succinate-d2 (30 mM in water, %P(13C) = 25.8 ± 5.1% (when produced) and %P(13C) = 14.2 ± 0.7% (when imaged), T1 = 74 ± 3 s), and proton images were recorded using (1)H hyperpolarized pyridine (100 mM in methanol-d4, %P(H) = 0.1 ± 0.02% (when imaged), T1 = 11 ± 0.1 s). Both contrast agents were hyperpolarized using parahydrogen (>90% para-fraction) in an automated 5.75 mT parahydrogen induced polarization (PHIP) hyperpolarizer. A magnetized path was demonstrated for successful transportation of a (13)C hyperpolarized contrast agent (1-(13)C-succinate-d2, sensitive to fast depolarization when at the Earth's magnetic field) from the PHIP polarizer to the 47.5 mT low-field MRI. While future polarizing and low-field MRI hardware and imaging sequence developments can further improve the low-field detection sensitivity, the current results demonstrate that microscale molecular imaging in vivo is already feasible at low (<50 mT) fields and potentially at low (~1 mM) metabolite concentrations.

  10. Concentric Rings K-Space Trajectory for Hyperpolarized 13C MR Spectroscopic Imaging

    PubMed Central

    Jiang, Wenwen; Lustig, Michael; Larson, Peder E.Z.

    2014-01-01

    Purpose To develop a robust and rapid imaging technique for hyperpolarized 13C MR Spectroscopic Imaging (MRSI) and investigate its performance. Methods A concentric rings readout trajectory with constant angular velocity is proposed for hyperpolarized 13C spectroscopic imaging and its properties are analyzed. Quantitative analyses of design tradeoffs are presented for several imaging scenarios. The first application of concentric rings on 13C phantoms and in vivo animal hyperpolarized 13C MRSI studies were performed to demonstrate the feasibility of the proposed method. Finally, a parallel imaging accelerated concentric rings study is presented. Results The concentric rings MRSI trajectory has the advantages of acquisition timesaving compared to echo-planar spectroscopic imaging (EPSI). It provides sufficient spectral bandwidth with relatively high SNR efficiency compared to EPSI and spiral techniques. Phantom and in vivo animal studies showed good image quality with half the scan time and reduced pulsatile flow artifacts compared to EPSI. Parallel imaging accelerated concentric rings showed advantages over Cartesian sampling in g-factor simulations and demonstrated aliasing-free image quality in a hyperpolarized 13C in vivo study. Conclusion The concentric rings trajectory is a robust and rapid imaging technique that fits very well with the speed, bandwidth, and resolution requirements of hyperpolarized 13C MRSI. PMID:25533653

  11. Nitric oxide: Mediator of nonadrenergic noncholinergic hyperpolarization of opossum esophageal circular smooth muscle

    SciTech Connect

    Du, C.; Murray, J.; Conklin, J.L.; Bates, J.N. )

    1991-03-15

    The electromyogram recorded from circular smooth muscle (SM) of opossum esophagus, either during peristalsis or when the intrinsic esophageal nerves are stimulated by an electrical field (EFS), consists of a hyperpolarization followed by a depolarization. This membrane response results from the interaction of a nonadrenergic-noncholinergic (NANC) neurotransmitter with its receptors on SM membrane. N{sup G}-nitro-L-arginine (L-NNA), an inhibitor of nitric oxide synthase, and nitric oxide (NO) were used to test the hypothesis that NO is a mediator of this NANC nerve-induced response. The transmembrane potential difference of circular SM cells of opossum esophagus was recorded with glass microelectrode. The nerve-mediated membrane response was evoked by EFS. L-NNA (50uM) abolished the initial hyperpolarization and reduced the amplitude of and the time to maximal depolarization. L-arginine (1mM), the substrate for NO synthase, antagonized the effect of L-NNA. Neither L-NNA nor L-arginine altered the resting membrane potential. Exogenous NO produced hyperpolarization of SM membrane potential and attenuated the amplitude of EFS-induced hyperpolarization and depolarization. Nitrosocysteine, a NO-containing compound, also hyperpolarized the membrane potential. Effect of NO was neither blocked by L-NNA nor by TTX. The data support the hypothesis that NO or an NO-containing compound mediates NANC nerve-induced responses of the esophageal SM membrane.

  12. Accommodation to hyperpolarizing currents: differences between motor and sensory nerves in mice.

    PubMed

    Nodera, Hiroyuki; Rutkove, Seward B

    2012-06-19

    Peripheral motor nerves have revealed variability in excitability by hyperpolarizing current at specific target response levels, likely reflecting differences in the hyperpolarization-activated current (Ih). Whether such variability in Ih exists in sensory axons is yet to be established. We performed nerve excitability testing in mouse tail motor and sensory nerves at 3 target response levels (20, 40, and 60% of the maximum amplitudes). Target-level dependent variability was present by long hyperpolarizing currents in motor and sensory nerves in which the recording at the low target level showed smaller threshold changes than at the high target level. Other excitability measures, however, showed no variability. Furthermore, the accommodation by long, strong hyperpolarization revealed smaller S3 accommodation (threshold change between the maximum and at the end of the 200 ms conditioning pulse) at the low target response level in sensory axons, but not in motor axons. Variation in the kinetics of the subtypes of the hyperpolarization-activated cyclic nucleotide-gated (HCN) channels in motor and sensory axons is the most likely explanation for these findings. The present study has proposed that nerve excitability testing may provide a non-invasive means for the assessment of the different types of Ih in neurological disorders where HCN subtypes play unique pathophysiological roles.

  13. Efficient Synthesis of Nicotinamide-1-15N for Ultrafast NMR Hyperpolarization Using Parahydrogen

    PubMed Central

    2016-01-01

    Nicotinamide (a vitamin B3 amide) is one of the key vitamins as well as a drug for treatment of M. tuberculosis, HIV, cancer, and other diseases. Here, an improved Zincke reaction methodology is presented allowing for straightforward and scalable synthesis of nicotinamide-1-15N with an excellent isotopic purity (98%) and good yield (55%). 15N nuclear spin label in nicotinamide-1-15N can be NMR hyperpolarized in seconds using parahydrogen gas. NMR hyperpolarization using the process of temporary conjugation between parahydrogen and to-be-hyperpolarized biomolecule on hexacoordinate iridium complex via the Signal Amplification By Reversible Exchange (SABRE) method significantly increases detection sensitivity (e.g., >20 000-fold for nicotinamide-1-15N at 9.4 T) as has been shown by Theis T. et al. (J. Am. Chem. Soc.2015, 137, 1404), and hyperpolarized in this fashion, nicotinamide-1-15N can be potentially used to probe metabolic processes in vivo in future studies. Moreover, the presented synthetic methodology utilizes mild reaction conditions, and therefore can also be potentially applied to synthesis of a wide range of 15N-enriched N-heterocycles that can be used as hyperpolarized contrast agents for future in vivo molecular imaging studies. PMID:26999571

  14. A Leucine Zipper Motif Essential for Gating of Hyperpolarization-activated Channels*

    PubMed Central

    Wemhöner, Konstantin; Silbernagel, Nicole; Marzian, Stefanie; Netter, Michael F.; Rinné, Susanne; Stansfeld, Phillip J.; Decher, Niels

    2012-01-01

    Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are pacemakers in cardiac myocytes and neurons. Although their membrane topology closely resembles that of voltage-gated K+ channels, the mechanism of their unique gating behavior in response to hyperpolarization is still poorly understood. We have identified a highly conserved leucine zipper motif in the S5 segment of HCN family members. In order to study the role of this motif for channel function, the leucine residues of the zipper were individually mutated to alanine, arginine, or glutamine residues. Leucine zipper mutants traffic to the plasma membrane, but the channels lose their sensitivity to open upon hyperpolarization. Thus, our data indicate that the leucine zipper is an important molecular determinant for hyperpolarization-activated channel gating. Residues of the leucine zipper interact with the adjacent S6 segment of the channel. This interaction is essential for voltage-dependent gating of the channel. The lower part of the leucine zipper, at the intracellular mouth of the channel, is important for stabilizing the closed state. Mutations at these sites increase current amplitudes or result in channels with deficient closing and increased min-Po. Our data are further supported by homology models of the open and closed state of the HCN2 channel pore. Thus, we conclude that the leucine zipper of HCN channels is a major determinant for hyperpolarization-activated channel gating. PMID:23048023

  15. Cortical hyperpolarization-activated depolarizing current takes part in the generation of focal paroxysmal activities

    PubMed Central

    Timofeev, Igor; Bazhenov, Maxim; Sejnowski, Terrence; Steriade, Mircea

    2002-01-01

    During paroxysmal neocortical oscillations, sudden depolarization leading to the next cycle occurs when the majority of cortical neurons are hyperpolarized. Both the Ca2+-dependent K+ currents (IK(Ca)) and disfacilitation play critical roles in the generation of hyperpolarizing potentials. In vivo experiments and computational models are used here to investigate whether the hyperpolarization-activated depolarizing current (Ih) in cortical neurons also contributes to the generation of paroxysmal onsets. Hyperpolarizing current pulses revealed a depolarizing sag in ≈20% of cortical neurons. Intracellular recordings from glial cells indirectly indicated an increase in extracellular potassium concentration ([K+]o) during paroxysmal activities, leading to a positive shift in the reversal potential of K+-mediated currents, including Ih. In the paroxysmal neocortex, ≈20% of neurons show repolarizing potentials originating from hyperpolarizations associated with depth-electroencephalogram positive waves of spike-wave complexes. The onset of these repolarizing potentials corresponds to maximal [K+]o as estimated from dual simultaneous impalements from neurons and glial cells. Computational models showed how, after the increased [K+]o, the interplay between Ih, IK(Ca), and a persistent Na+ current, INa(P), could organize paroxysmal oscillations at a frequency of 2–3 Hz. PMID:12089324

  16. Real-time assessment of Krebs cycle metabolism using hyperpolarized 13C magnetic resonance spectroscopy.

    PubMed

    Schroeder, Marie A; Atherton, Helen J; Ball, Daniel R; Cole, Mark A; Heather, Lisa C; Griffin, Julian L; Clarke, Kieran; Radda, George K; Tyler, Damian J

    2009-08-01

    The Krebs cycle plays a fundamental role in cardiac energy production and is often implicated in the energetic imbalance characteristic of heart disease. In this study, we measured Krebs cycle flux in real time in perfused rat hearts using hyperpolarized magnetic resonance spectroscopy (MRS). [2-(13)C]Pyruvate was hyperpolarized and infused into isolated perfused hearts in both healthy and postischemic metabolic states. We followed the enzymatic conversion of pyruvate to lactate, acetylcarnitine, citrate, and glutamate with 1 s temporal resolution. The appearance of (13)C-labeled glutamate was delayed compared with that of other metabolites, indicating that Krebs cycle flux can be measured directly. The production of (13)C-labeled citrate and glutamate was decreased postischemia, as opposed to lactate, which was significantly elevated. These results showed that the control and fluxes of the Krebs cycle in heart disease can be studied using hyperpolarized [2-(13)C]pyruvate.

  17. Hyperpolarized 13C pyruvate mouse brain metabolism with absorptive-mode EPSI at 1 T

    NASA Astrophysics Data System (ADS)

    Miloushev, Vesselin Z.; Di Gialleonardo, Valentina; Salamanca-Cardona, Lucia; Correa, Fabian; Granlund, Kristin L.; Keshari, Kayvan R.

    2017-02-01

    The expected signal in echo-planar spectroscopic imaging experiments was explicitly modeled jointly in spatial and spectral dimensions. Using this as a basis, absorptive-mode type detection can be achieved by appropriate choice of spectral delays and post-processing techniques. We discuss the effects of gradient imperfections and demonstrate the implementation of this sequence at low field (1.05 T), with application to hyperpolarized [1-13C] pyruvate imaging of the mouse brain. The sequence achieves sufficient signal-to-noise to monitor the conversion of hyperpolarized [1-13C] pyruvate to lactate in the mouse brain. Hyperpolarized pyruvate imaging of mouse brain metabolism using an absorptive-mode EPSI sequence can be applied to more sophisticated murine disease and treatment models. The simple modifications presented in this work, which permit absorptive-mode detection, are directly translatable to human clinical imaging and generate improved absorptive-mode spectra without the need for refocusing pulses.

  18. Decomposition of cumyl hydroperoxide in the presence of sulphonated silica in a flow-type system

    SciTech Connect

    Shelpakova, N.A.; Ioffa, A.F.

    1993-12-31

    An investigation has been made of the decomposition of cumyl hydroperoxide (CHP) in the presence of silica-based sulphocationites in a flow-type system. It was established that the given specimens are effective catalysts for the decomposition of CHP into phenol and acetone. It was shown that, in the course of the process, no irreversible poisoning of the surface of the catalyst by the products of CHP decomposition occurs. Data of chromatographic analysis of the products of CHP decomposition in the presence of sulphuric acid and silica-based sulphocationites are given.

  19. Hyperpolarized Xenon-129 Magnetic Resonance Imaging of Functional Lung Microstructure

    NASA Astrophysics Data System (ADS)

    Dregely, Isabel

    Hyperpolarized 129Xe (HXe) is a non-invasive contrast agent for lung magnetic resonance imaging (MRI), which upon inhalation follows the functional pathway of oxygen in the lung by dissolving into lung tissue structures and entering the blood stream. HXe MRI therefore provides unique opportunities for functional lung imaging of gas exchange which occurs from alveolar air spaces across the air-blood boundary into parenchymal tissue. However challenges in acquisition speed and signal-to-noise ratio have limited the development of a HXe imaging biomarker to diagnose lung disease. This thesis addresses these challenges by introducing parallel imaging to HXe MRI. Parallel imaging requires dedicated hardware. This work describes design, implementation, and characterization of a 32-channel phased-array chest receive coil with an integrated asymmetric birdcage transmit coil tuned to the HXe resonance on a 3 Tesla MRI system. Using the newly developed human chest coil, a functional HXe imaging method, multiple exchange time xenon magnetization transfer contrast (MXTC) is implemented. MXTC dynamically encodes HXe gas exchange into the image contrast. This permits two parameters to be derived regionally which are related to gas-exchange functionality by characterizing tissue-to-alveolar-volume ratio and alveolar wall thickness in the lung parenchyma. Initial results in healthy subjects demonstrate the sensitivity of MXTC by quantifying the subtle changes in lung microstructure in response to orientation and lung inflation. Our results in subjects with lung disease show that the MXTC-derived functional tissue density parameter exhibits excellent agreement with established imaging techniques. The newly developed dynamic parameter, which characterizes the alveolar wall, was elevated in subjects with lung disease, most likely indicating parenchymal inflammation. In light of these observations we believe that MXTC has potential as a biomarker for the regional quantification of 1

  20. Magnetic resonance imaging with hyperpolarized agents: methods and applications.

    PubMed

    Adamson, Erin; Ludwig, Kai; Mummy, David; Fain, Sean B

    2017-04-06

    In the past decade, hyperpolarized (HP) contrast agents have been under active development for MRI applications to address the twin challenges of functional and quantitative imaging. Both HP helium (3He) and xenon (129Xe) gases have reached the stage where they are under study in clinical research. HP 129Xe, in particular, is poised for larger scale clinical research to investigate asthma, chronic obstructive pulmonary disease, and fibrotic lung diseases. With advances in polarizer technology and unique capabilities for imaging of 129Xe gas exchange into lung tissue and blood, HP 129Xe MRI is attracting new attention. In parallel, HP 13C and 15N MRI methods have steadily advanced in a wide range of pre-clinical research applications for imaging metabolism in various cancers and cardiac disease. The HP [1-13C] pyruvate MRI technique, in particular, has undergone phase I trials in prostate cancer and is poised for investigational new drug trials at multiple institutions in cancer and cardiac applications. This review treats the methodology behind both HP gases and HP 13C and 15N liquid state agents. Gas and liquid phase HP agents share similar technologies for achieving non-equilibrium polarization outside the field of the MRI scanner, strategies for image data acquisition, and translational challenges in moving from pre-clinical to clinical research. To cover the wide array of methods and applications, this review is organized by numerical section into 1) a brief introduction, 2) the physical and biological properties of the most common polarized agents with a brief summary of applications and methods of polarization, 3) methods for image acquisition and reconstruction specific to improving data acquisition efficiency for HP MRI, 4) the main physical properties that enable unique measures of physiology or metabolic pathways, followed by a more detailed

  1. Ion Permeabilities in Mouse Sperm Reveal an External Trigger for SLO3-Dependent Hyperpolarization

    PubMed Central

    Chávez, Julio C.; de la Vega-Beltrán, José L.; Escoffier, Jessica; Visconti, Pablo E.; Treviño, Claudia L.; Darszon, Alberto; Salkoff, Lawrence; Santi, Celia M.

    2013-01-01

    Unlike most cells of the body which function in an ionic environment controlled within narrow limits, spermatozoa must function in a less controlled external environment. In order to better understand how sperm control their membrane potential in different ionic conditions, we measured mouse sperm membrane potentials under a variety of conditions and at different external K+ concentrations, both before and after capacitation. Experiments were undertaken using both wild-type, and mutant mouse sperm from the knock-out strain of the sperm-specific, pH-sensitive, SLO3 K+ channel. Membrane voltage data were fit to the Goldman-Hodgkin-Katz equation. Our study revealed a significant membrane permeability to both K+ and Cl− before capacitation, as well as Na+. The permeability to both K+ and Cl− has the effect of preventing large changes in membrane potential when the extracellular concentration of either ion is changed. Such a mechanism may protect against undesired shifts in membrane potential in changing ionic environments. We found that a significant portion of resting membrane potassium permeability in wild-type sperm was contributed by SLO3 K+ channels. We also found that further activation of SLO3 channels was the essential mechanism producing membrane hyperpolarization under two separate conditions, 1) elevation of external pH prior to capacitation and 2) capacitating conditions. Both conditions produced a significant membrane hyperpolarization in wild-type which was absent in SLO3 mutant sperm. Hyperpolarization in both conditions may result from activation of SLO3 channels by raising intracellular pH; however, demonstrating that SLO3-dependent hyperpolarization is achieved by an alkaline environment alone shows that SLO3 channel activation might occur independently of other events associated with capacitation. For example sperm may undergo stages of membrane hyperpolarization when reaching alkaline regions of the female genital tract. Significantly, other

  2. Single shot three‐dimensional pulse sequence for hyperpolarized 13C MRI

    PubMed Central

    Wang, Jiazheng; Wright, Alan J.; Hu, De‐en; Hesketh, Richard

    2016-01-01

    Purpose Metabolic imaging with hyperpolarized 13C‐labeled cell substrates is a promising technique for imaging tissue metabolism in vivo. However, the transient nature of the hyperpolarization, and its depletion following excitation, limits the imaging time and the number of excitation pulses that can be used. We describe here a single‐shot three‐dimensional (3D) imaging sequence and demonstrate its capability to generate 13C MR images in tumor‐bearing mice injected with hyperpolarized [1‐13C]pyruvate. Methods The pulse sequence acquires a stack‐of‐spirals at two spin echoes after a single excitation pulse and encodes the kz‐dimension in an interleaved manner to enhance robustness to B0 inhomogeneity. Spectral‐spatial pulses are used to acquire dynamic 3D images from selected hyperpolarized 13C‐labeled metabolites. Results A nominal spatial/temporal resolution of 1.25 × 1.25 × 2.5 mm3 × 2 s was achieved in tumor images of hyperpolarized [1‐13C]pyruvate and [1‐13C]lactate acquired in vivo. Higher resolution in the z‐direction, with a different k‐space trajectory, was demonstrated in measurements on a thermally polarized [1‐13C]lactate phantom. Conclusion The pulse sequence is capable of imaging hyperpolarized 13C‐labeled substrates at relatively high spatial and temporal resolutions and is robust to moderate system imperfections. Magn Reson Med 77:740–752, 2017. © 2016 The Authors Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. PMID:26916384

  3. Mouse Sperm Membrane Potential Hyperpolarization Is Necessary and Sufficient to Prepare Sperm for the Acrosome Reaction*

    PubMed Central

    De La Vega-Beltran, Jose Luis; Sánchez-Cárdenas, Claudia; Krapf, Darío; Hernandez-González, Enrique O.; Wertheimer, Eva; Treviño, Claudia L.; Visconti, Pablo E.; Darszon, Alberto

    2012-01-01

    Mammalian sperm are unable to fertilize the egg immediately after ejaculation; they acquire this capacity during migration in the female reproductive tract. This maturational process is called capacitation and in mouse sperm it involves a plasma membrane reorganization, extensive changes in the state of protein phosphorylation, increases in intracellular pH (pHi) and Ca2+ ([Ca2+]i), and the appearance of hyperactivated motility. In addition, mouse sperm capacitation is associated with the hyperpolarization of the cell membrane potential. However, the functional role of this process is not known. In this work, to dissect the role of this membrane potential change, hyperpolarization was induced in noncapacitated sperm using either the ENaC inhibitor amiloride, the CFTR agonist genistein or the K+ ionophore valinomycin. In this experimental setting, other capacitation-associated processes such as activation of a cAMP-dependent pathway and the consequent increase in protein tyrosine phosphorylation were not observed. However, hyperpolarization was sufficient to prepare sperm for the acrosome reaction induced either by depolarization with high K+ or by addition of solubilized zona pellucida (sZP). Moreover, K+ and sZP were also able to increase [Ca2+]i in non-capacitated sperm treated with these hyperpolarizing agents but not in untreated cells. On the other hand, in conditions that support capacitation-associated processes blocking hyperpolarization by adding valinomycin and increasing K+ concentrations inhibited the agonist-induced acrosome reaction as well as the increase in [Ca2+]i. Altogether, these results suggest that sperm hyperpolarization by itself is key to enabling mice sperm to undergo the acrosome reaction. PMID:23095755

  4. Synthesis of Long-T1 Silicon Nanoparticles for Hyperpolarized 29Si Magnetic Resonance Imaging

    PubMed Central

    Atkins, Tonya M.; Cassidy, Maja C.; Lee, Menyoung; Ganguly, Shreyashi; Marcus, Charles M.; Kauzlarich, Susan M.

    2013-01-01

    We describe the synthesis, materials characterization and dynamic nuclear polarization (DNP) of amorphous and crystalline silicon nanoparticles for use as hyperpolarized magnetic resonance imaging (MRI) agents. The particles were synthesized by means of a metathesis reaction between sodium silicide (Na4Si4) and silicon tetrachloride (SiCl4) and were surface functionalized with a variety of passivating ligands. The synthesis scheme results in particles of diameter ~10 nm with long size-adjusted 29Si spin lattice relaxation (T1) times (> 600 s), which are retained after hyperpolarization by low temperature DNP. PMID:23350651

  5. Chemistry and biochemistry of 13C hyperpolarized magnetic resonance using dynamic nuclear polarization

    PubMed Central

    Keshari, Kayvan R.; Wilson, David M.

    2014-01-01

    The study of transient chemical phenomena by conventional NMR has proved elusive, particularly for non-1H nuclei. For 13C, hyperpolarization using the dynamic nuclear polarization (DNP) technique has emerged as a powerful means to improve SNR. The recent development of rapid dissolution DNP methods has facilitated previously impossible in vitro and in vivo study of small molecules. This review presents the basics of the DNP technique, identification of appropriate DNP substrates, and approaches to increase hyperpolarized signal lifetimes. Also addressed are the biochemical events to which DNP-NMR has been applied, with descriptions of several probes that have met with in vivo success. PMID:24363044

  6. Production of hyperpolarized xenon in a static pump cell: Numerical simulations and experiments

    SciTech Connect

    Fink, A.; Baumer, D.; Brunner, E.

    2005-11-15

    Hyperpolarized {sup 129}Xe finds numerous applications in NMR spectroscopy and magnetic resonance imaging. The production of hyperpolarized {sup 129}Xe by spin-exchange optical pumping is therefore an important experimental issue. We model the three-dimensional transport processes within a so-called batch mode pump cell via numerical finite element method simulations and compare the results with experimental data. In particular, the influence of different experimental parameters, such as temperature, xenon and nitrogen partial pressure, laser power, and radius-to-length ratio of a cylindrical pump cell, is evaluated. The developed numerical method is capable of describing the spin-exchange optical pumping process in a realistic manner.

  7. Characterization of a chloride conductance activated by hyperpolarization in Aplysia neurones.

    PubMed

    Chesnoy-Marchais, D

    1983-09-01

    A voltage-clamp study was made of some properties of the non-synaptic hyperpolarization-activated Cl- conductance recently described in Aplysia neurones loaded with Cl- ions (Chesnoy-Marchais, 1982). The experiments were performed on an identified family of neurones, which present cholinergic responses allowing an easy measurement of the equilibrium potentials of Cl- (ECl) and K+ ions (EK). The Cl- selectivity of the hyperpolarization-activated conductance was deduced from four observations: (1) the extrapolated reversal potential of the hyperpolarization-activated current, Er, was close to the reversal potential of the cholinergic Cl- response, which is the equilibrium potential for Cl- ions, ECl. (2) Modifications of the intracellular or extracellular Cl- concentration induced changes of the reversal potential Er. (3) A prolonged and intense activation of the current lowered the intracellular Cl- concentration. (4) The current persisted after complete substitution of intracellular and extracellular cations by CS+ ions, as well as after replacement of extracellular Na+ ions by Tris. The steady-state Cl- conductance (gss) increases steeply with hyperpolarization. The kinetics of activation and deactivation are exponential and are characterized by the same voltage-dependent time constant (tau), of the order of a few seconds or fractions of seconds. The curves gss(V) and tau (V) can both be fitted by a two-state model in which the rate constants are exponential functions of the membrane potential (e-fold change for 12-16 mV). The Cl- current is much more affected by changes of the intracellular Cl- concentration than predicted simply from the change in Cl- driving force. Both the conductance and the time constant of activation are strongly modified. Modifications of the extracellular Cl- concentration do not always alter the amplitude of the hyperpolarization-activated Cl- current, but systematically affect its kinetics. The hyperpolarization-activated current is

  8. Effect of ionic interaction between a hyperpolarized magnetic resonance chemical probe and a gadolinium contrast agent for the hyperpolarized lifetime after dissolution

    NASA Astrophysics Data System (ADS)

    Takakusagi, Yoichi; Inoue, Kaori; Naganuma, Tatsuya; Hyodo, Fuminori; Ichikawa, Kazuhiro

    2016-09-01

    In hyperpolarization of 13C-enriched magnetic resonance chemical probes in the solid-state, a trace amount of gadolinium (Gd) contrast agent can be used to maximize polarization of the 13C nuclear spins. Here, we report systematic measurement of the spin-lattice relaxation time (T1) and enhancement level of 13C-enriched chemical probes in the presence of various Gd contrast agents in the liquid-state after dissolution. Using two different 13C probes having opposite electric charges at neutral pH, we clearly show the T1 of hyperpolarized 13C was barely affected by the use of a Gd complex that displays repulsive interaction with the 13C probe in solution, whilst T1 was drastically shortened when there was ionic attraction between probe and complex.

  9. Developmental conditioning of endothelium-derived hyperpolarizing factor-mediated vasorelaxation

    PubMed Central

    Stead, Rebecca; Musa, Moji G.; Bryant, Claire L.; Lanham, Stuart A.; Johnston, David A.; Reynolds, Richard; Torrens, Christopher; Fraser, Paul A.; Clough, Geraldine F.

    2016-01-01

    Objectives: The endothelium maintains vascular homeostasis through the release of endothelium-derived relaxing factors (EDRF) and endothelium-derived hyperpolarization (EDH). The balance in EDH : EDRF is disturbed in cardiovascular disease and may also be susceptible to developmental conditioning through exposure to an adverse uterine environment to predispose to later risk of hypertension and vascular disease. Methods: Developmentally conditioned changes in EDH : EDRF signalling pathways were investigated in cremaster arterioles (18–32 μm diameter) and third-order mesenteric arteries of adult male mice offspring of dams fed either a fat-rich (high fat, HF, 45% energy from fat) or control (C, 10% energy from fat) diet. After weaning, offspring either continued on high fat or were placed on control diets to give four dietary groups (C/C, HF/C, C/HF, and HF/HF) and studied at 15 weeks of age. Results: EDH via intermediate (IKCa) and small (SKca) conductance calcium-activated potassium channels contributed less than 10% to arteriolar acetylcholine-induced relaxation in in-situ conditioned HF/C offspring compared with ∼60% in C/C (P < 0.01). The conditioned reduction in EDH signalling in HF/C offspring was reversed in offspring exposed to a high-fat diet both before and after weaning (HF/HF, 55%, P < 0.01 vs. HF/C). EDH signalling was unaffected in arterioles from C/HF offspring. The changes in EDH : EDRF were associated with altered endothelial cell expression and localization of IKCa channels. Conclusion: This is the first evidence that EDH-mediated microvascular relaxation is susceptible to an adverse developmental environment through down-regulation of the IKCa signalling pathway. Conditioned offspring exposed to a ‘second hit’ (HF/HF) exhibit adaptive vascular mechanisms to preserve dilator function. PMID:26682783

  10. Multi-point optimization of recirculation flow type casing treatment in centrifugal compressors

    NASA Astrophysics Data System (ADS)

    Tun, Min Thaw; Sakaguchi, Daisaku

    2016-06-01

    High-pressure ratio and wide operating range are highly required for a turbocharger in diesel engines. A recirculation flow type casing treatment is effective for flow range enhancement of centrifugal compressors. Two ring grooves on a suction pipe and a shroud casing wall are connected by means of an annular passage and stable recirculation flow is formed at small flow rates from the downstream groove toward the upstream groove through the annular bypass. The shape of baseline recirculation flow type casing is modified and optimized by using a multi-point optimization code with a metamodel assisted evolutionary algorithm embedding a commercial CFD code CFX from ANSYS. The numerical optimization results give the optimized design of casing with improving adiabatic efficiency in wide operating flow rate range. Sensitivity analysis of design parameters as a function of efficiency has been performed. It is found that the optimized casing design provides optimized recirculation flow rate, in which an increment of entropy rise is minimized at grooves and passages of the rotating impeller.

  11. In Vivo Lung Morphometry with Accelerated Hyperpolarized 3He Diffusion MRI: A Preliminary Study

    PubMed Central

    Chang, Yulin V.; Quirk, James D.; Yablonskiy, Dmitriy A.

    2014-01-01

    Purpose Parallel imaging can be used to reduce imaging time and to increase the spatial coverage in hyperpolarized gas MRI of the lung. In this proof-of-concept study we investigate the effects of parallel imaging on the morphometric measurement of lung microstructure using diffusion MRI with hyperpolarized 3He. Methods Fully sampled and under-sampled multi-b diffusion data were acquired from human subjects using an 8-channel 3He receive coil. A parallel imaging reconstruction technique (GRAPPA) was used to reconstruct under-sampled k-space data. The morphometric results of the GRAPPA-reconstructed data were compared with the results of fully sampled data for three types of subjects: healthy volunteers, mild, and moderate COPD patients. Results Morphometric measurements varied only slightly at mild acceleration factors. The results were largely well preserved compared to fully sampled data for different lung conditions. Conclusion Parallel imaging, given sufficient signal-to-noise ratio, provides a reliable means to accelerate hyperpolarized-gas MRI with no significant difference in the measurement of lung morphometry from the fully sampled images. GRAPPA is a promising technique to significantly reduce imaging time and/or to improve the spatial coverage for the morphometric measurement with hyperpolarized gases. PMID:24799044

  12. Acetylcholine releases endothelium-derived hyperpolarizing factor and EDRF from rat blood vessels.

    PubMed Central

    Chen, G.; Suzuki, H.; Weston, A. H.

    1988-01-01

    1. The effects of haemoglobin and methylene blue on the acetylcholine (ACh)-induced electrical and mechanical responses of smooth muscle cells were investigated in rat aorta and rat main pulmonary artery. 2. When the endothelium was intact, ACh induced a transient hyperpolarization and sustained relaxation of tissues precontracted with noradrenaline. Both hyperpolarization and relaxation were absent in preparations without endothelium. 3. Haemoglobin and methylene blue inhibited the ACh-induced relaxation, but not the transient hyperpolarization. 4. In aorta with an intact endothelium, ACh produced an increase in both the rate of 86Rb efflux and tissue cyclic GMP levels. The changes in ion flux were unaffected by either haemoglobin or methylene blue in concentrations which almost abolished the increase in cyclic GMP concentrations. 5. In arteries with an intact endothelium, indomethacin had no effect on the ACh-induced electrical and mechanical responses or on the increase in 86Rb efflux and tissue cyclic GMP levels. 6. It is concluded that in the rat aorta and rat main pulmonary artery, ACh releases two different substances, an endothelium-derived relaxing factor (EDRF) and a hyperpolarizing factor (EDHF), from the endothelial cells. Neither substance appears to be derived from a pathway dependent on cyclo-oxygenase. EDHF seems to play a minor role in the relaxation of noradrenaline-induced contractions. PMID:2851359

  13. Imaging Renal Urea Handling in Rats at Millimeter Resolution using Hyperpolarized Magnetic Resonance Relaxometry

    PubMed Central

    Reed, Galen D.; von Morze, Cornelius; Verkman, Alan S.; Koelsch, Bertram L.; Chaumeil, Myriam M.; Lustig, Michael; Ronen, Sabrina M.; Bok, Robert A.; Sands, Jeff M.; Larson, Peder E. Z.; Wang, Zhen J.; Larsen, Jan Henrik Ardenkjær; Kurhanewicz, John; Vigneron, Daniel B.

    2016-01-01

    In vivo spin spin relaxation time (T2) heterogeneity of hyperpolarized [13C,15N2]urea in the rat kidney was investigated. Selective quenching of the vascular hyperpolarized 13C signal with a macromolecular relaxation agent revealed that a long-T2 component of the [13C,15N2]urea signal originated from the renal extravascular space, thus allowing the vascular and renal filtrate contrast agent pools of the [13C,15N2]urea to be distinguished via multi-exponential analysis. The T2 response to induced diuresis and antidiuresis was performed with two imaging agents: hyperpolarized [13C,15N2]urea and a control agent hyperpolarized bis-1,1-(hydroxymethyl)-1-13C-cyclopropane-2H8. Large T2 increases in the inner-medullar and papilla were observed with the former agent and not the latter during antidiuresis. Therefore, [13C,15N2]urea relaxometry is sensitive to two steps of the renal urea handling process: glomerular filtration and the inner-medullary urea transporter (UT)-A1 and UT-A3 mediated urea concentrating process. Simple motion correction and subspace denoising algorithms are presented to aid in the multi exponential data analysis. Furthermore, a T2-edited, ultra long echo time sequence was developed for sub-2 mm3 resolution 3D encoding of urea by exploiting relaxation differences in the vascular and filtrate pools. PMID:27570835

  14. An optimized microfabricated platform for the optical generation and detection of hyperpolarized 129Xe

    PubMed Central

    Kennedy, Daniel J.; Seltzer, Scott J.; Jiménez-Martínez, Ricardo; Ring, Hattie L.; Malecek, Nicolas S.; Knappe, Svenja; Donley, Elizabeth A.; Kitching, John; Bajaj, Vikram S.; Pines, Alexander

    2017-01-01

    Low thermal-equilibrium nuclear spin polarizations and the need for sophisticated instrumentation render conventional nuclear magnetic resonance (NMR) spectroscopy and imaging (MRI) incompatible with small-scale microfluidic devices. Hyperpolarized 129Xe gas has found use in the study of many materials but has required very large and expensive instrumentation. Recently a microfabricated device with modest instrumentation demonstrated all-optical hyperpolarization and detection of 129Xe gas. This device was limited by 129Xe polarizations less than 1%, 129Xe NMR signals smaller than 20 nT, and transport of hyperpolarized 129Xe over millimeter lengths. Higher polarizations, versatile detection schemes, and flow of 129Xe over larger distances are desirable for wider applications. Here we demonstrate an ultra-sensitive microfabricated platform that achieves 129Xe polarizations reaching 7%, NMR signals exceeding 1 μT, lifetimes up to 6 s, and simultaneous two-mode detection, consisting of a high-sensitivity in situ channel with signal-to-noise of 105 and a lower-sensitivity ex situ detection channel which may be useful in a wider variety of conditions. 129Xe is hyperpolarized and detected in locations more than 1 cm apart. Our versatile device is an optimal platform for microfluidic magnetic resonance in particular, but equally attractive for wider nuclear spin applications benefitting from ultra-sensitive detection, long coherences, and simple instrumentation. PMID:28266629

  15. A hyperpolarization-activated inward current alters swim frequency of the pteropod mollusk Clione limacina.

    PubMed

    Pirtle, Thomas J; Willingham, Kyle; Satterlie, Richard A

    2010-12-01

    The pteropod mollusk, Clione limacina, exhibits behaviorally relevant swim speed changes that occur within the context of the animal's ecology. Modulation of C. limacina swimming speed involves changes that occur at the network and cellular levels. Intracellular recordings from interneurons of the swim central pattern generator show the presence of a sag potential that is indicative of the hyperpolarization-activated inward current (I(h)). Here we provide evidence that I(h) in primary swim interneurons plays a role in C. limacina swimming speed control and may be a modulatory target. Recordings from central pattern generator swim interneurons show that hyperpolarizing current injection produces a sag potential that lasts for the duration of the hyperpolarization, a characteristic of cells possessing I(h). Following the hyperpolarizing current injection, swim interneurons also exhibit postinhibitory rebound (PIR). Serotonin enhances the sag potential of C. limacina swim interneurons while the I(h) blocker, ZD7288, reduces the sag potential. Furthermore, a negative correlation was found between the amplitude of the sag potential and latency to PIR. Because latency to PIR was previously shown to influence swimming speed, we hypothesize that I(h) has an effect on swimming speed. The I(h) blocker, ZD7288, suppresses swimming in C. limacina and inhibits serotonin-induced acceleration, evidence that supports our hypothesis.

  16. An optimized microfabricated platform for the optical generation and detection of hyperpolarized 129Xe

    NASA Astrophysics Data System (ADS)

    Kennedy, Daniel J.; Seltzer, Scott J.; Jiménez-Martínez, Ricardo; Ring, Hattie L.; Malecek, Nicolas S.; Knappe, Svenja; Donley, Elizabeth A.; Kitching, John; Bajaj, Vikram S.; Pines, Alexander

    2017-03-01

    Low thermal-equilibrium nuclear spin polarizations and the need for sophisticated instrumentation render conventional nuclear magnetic resonance (NMR) spectroscopy and imaging (MRI) incompatible with small-scale microfluidic devices. Hyperpolarized 129Xe gas has found use in the study of many materials but has required very large and expensive instrumentation. Recently a microfabricated device with modest instrumentation demonstrated all-optical hyperpolarization and detection of 129Xe gas. This device was limited by 129Xe polarizations less than 1%, 129Xe NMR signals smaller than 20 nT, and transport of hyperpolarized 129Xe over millimeter lengths. Higher polarizations, versatile detection schemes, and flow of 129Xe over larger distances are desirable for wider applications. Here we demonstrate an ultra-sensitive microfabricated platform that achieves 129Xe polarizations reaching 7%, NMR signals exceeding 1 μT, lifetimes up to 6 s, and simultaneous two-mode detection, consisting of a high-sensitivity in situ channel with signal-to-noise of 105 and a lower-sensitivity ex situ detection channel which may be useful in a wider variety of conditions. 129Xe is hyperpolarized and detected in locations more than 1 cm apart. Our versatile device is an optimal platform for microfluidic magnetic resonance in particular, but equally attractive for wider nuclear spin applications benefitting from ultra-sensitive detection, long coherences, and simple instrumentation.

  17. Milli-tesla NMR and spectrophotometry of liquids hyperpolarized by dissolution dynamic nuclear polarization

    NASA Astrophysics Data System (ADS)

    Zhu, Yue; Chen, Chia-Hsiu; Wilson, Zechariah; Savukov, Igor; Hilty, Christian

    2016-09-01

    Hyperpolarization methods offer a unique means of improving low signal strength obtained in low-field NMR. Here, simultaneous measurements of NMR at a field of 0.7 mT and laser optical absorption from samples hyperpolarized by dissolution dynamic nuclear polarization (D-DNP) are reported. The NMR measurement field closely corresponds to a typical field encountered during sample injection in a D-DNP experiment. The optical spectroscopy allows determination of the concentration of the free radical required for DNP. Correlation of radical concentration to NMR measurement of spin polarization and spin-lattice relaxation time allows determination of relaxivity and can be used for optimization of the D-DNP process. Further, the observation of the nuclear Overhauser effect originating from hyperpolarized spins is demonstrated. Signals from 1H and 19F in a mixture of trifluoroethanol and water are detected in a single spectrum, while different atoms of the same type are distinguished by J-coupling patterns. The resulting signal changes of individual peaks are indicative of molecular contact, suggesting a new application area of hyperpolarized low-field NMR for the determination of intermolecular interactions.

  18. Pomeranchuk cell for hyperpolarized 3He based on the brute force method

    NASA Astrophysics Data System (ADS)

    Makino, Seiji; Tanaka, Masayoshi; Ueda, Kunihiro; Fujiwara, Mamoru; Fujimura, Hisako; Yosoi, Masaru; Ohta, Takeshi; Frossati, Giorgio; de Waard, Arlette; Rouille, Gerard

    2014-09-01

    MRI (Magnetic Resonance Imaging) has been used for the medical diagnosis as a radiation-free imaging equipment. Since the proton has been mainly used for medical MRI, usefulness has been rather restrictive. As an example for expanding the range of applicability, MRI with hyperpolarized 3He gas has been used for the lung disease. Here, ``hyperpolarized'' means ``polarized higher than the thermal equilibrium polarization.'' For producing a large amount of hyperpolarized 3He gas at a time, we have been developing a hyperpolarization technique based on the brute force method which uses an ultralow temperature of a few mK and a strong magnetic field around 17 T in combination with the principle of the Pomeranchuk cooling. The Pomeranchuk cell made with non-metallic materials of small heat capacity is attached to the 3He/4He dilution refrigerator using a sintered silver allowing large heat conduction. After the sensors to monitor the temperature and pressure of 3He are calibrated and the Pomeranchuk cell is constructed, the system is tested. Then, the solidification of 3He and the measurement of NMR (Nuclear Magnetic Resonance) signals of 3He under the magnetic field of 17 T are carried out. The current status is reported in this talk.

  19. Hyperpolarized helium-3 mouse lung MRI: Studies of lung structure and function

    NASA Astrophysics Data System (ADS)

    Dugas, Joseph Paul

    Hyperpolarized 3He magnetic resonance imaging (MRI) of human and animal lungs has displayed promising and useful applications to studies of lung structure and function in both healthy and diseased lungs. Hyperpolarized 3He MRI allows the visualization of gas in the gas-exchange spaces of the lungs (as opposed to tissue) and has proven especially effective in studying diseases that are characterized by ventilation defects, such as emphysema. In particular, in-vivo measurements of the 3He apparent diffusion coefficient (ADC) can quantify lung structure by measuring its restrictive effects on the motion of 3He spins. This allows for detection and longitudinal tracking of changes in micro-architecture that result from disease destruction of alveolar walls. Due, in part, to the difficulties inherent in administering and imaging hyperpolarized 3He within the small (0.5 cc volume) mouse lung, applications of hyperpolarized 3He MRI techniques to laboratory mice are scarce. We have been able to implement and improve the techniques of hyperpolarized 3He mouse lung MRI and subsequently apply them to studies of several mouse models of disease, including elastase-induced emphysema, smoking-induced emphysema, and lung cancer. Here we detail the design, development, and implementation of a versatile, electronically-controlled, small animal ventilator that is capable of delivering tiny volumes of hyperpolarized 3He, mixed with oxygen, to the mouse and is also compatible with both the easily depolarized 3He gas and the highly magnetic environment within and around an imaging magnet. Also described are NM techniques developed to improve the signal-to-noise ratio of our images and effectively utilize the gas hyperpolarization. Applications of these technologies and techniques to small animal models of disease are presented wherein we have measured up to a 35% increase in 3He ADC in mice with elastase-induced emphysema as compared to healthy mice. We also demonstrate the potential

  20. Scalable control program for multiprecursor flow-type atomic layer deposition system

    SciTech Connect

    Selvaraj, Sathees Kannan; Takoudis, Christos G.

    2015-01-01

    The authors report the development and implementation of a scalable control program to control flow type atomic layer deposition (ALD) reactor with multiple precursor delivery lines. The program logic is written and tested in LABVIEW environment to control ALD reactor with four precursor delivery lines to deposit up to four layers of different materials in cyclic manner. The programming logic is conceived such that to facilitate scale up for depositing more layers with multiple precursors and scale down for using single layer with any one precursor in the ALD reactor. The program takes precursor and oxidizer exposure and purging times as input and controls the sequential opening and closing of the valves to facilitate the complex ALD process in cyclic manner. The program could be used to deposit materials from any single line or in tandem with other lines in any combination and in any sequence.

  1. Flow type barrier-discharge UV photoreactor for irradiating liquids and gases

    NASA Astrophysics Data System (ADS)

    Tsvetkov, V. M.; Pikulev, A. A.

    2012-04-01

    A flow type barrier-discharge UV photoreactor intended for irradiation of liquids and gases has been developed. In the proposed reactor design, both the discharge region and processed medium occur inside the bulb of an excilamp and the electrodes are made of an UV-reflecting material. The UV radiation intensity in the photoreactor is determined using the photochemical reaction of acetic acid decomposition (CH3COOH + hν → CH4 + CO2) that takes place under the action of KrCl exciplex emission at 222 nm, and the UV exposure dose is evaluated by measuring the evolved gas volume. The experiments showed that the UV radiation intensity in the proposed photoreactor is higher by an order of magnitude than that at the surface of an excilamp with analogous geometry.

  2. Hyperpolarizing inhibition develops without trophic support by GABA in cultured rat midbrain neurons.

    PubMed

    Titz, Stefan; Hans, Michael; Kelsch, Wolfgang; Lewen, Andrea; Swandulla, Dieter; Misgeld, Ulrich

    2003-08-01

    During a limited period of early neuronal development, GABA is depolarizing and elevates [Ca2+]i, which mediates the trophic action of GABA in neuronal maturation. We tested the attractive hypothesis that GABA itself promotes the developmental change of its response from depolarizing to hyperpolarizing (Ganguly et al. 2001). In cultured midbrain neurons we found that the GABA response changed from depolarizing to hyperpolarizing, although GABAA receptors had been blocked throughout development. In immature neurons prolonged exposure of the cells to nanomolar concentrations of GABA or brief repetitive applications of GABA strongly diminished the elevation of [Ca+]i by GABA. As revealed by gramicidin perforated-patch recording, reduced [Ca2+]i responses were due to a diminished driving force for Cl-. This suggests that immature neurons do not have an efficient inward transport that can compensate the loss of cytosolic Cl-resulting from sustained GABAA receptor activation by ambient GABA. Transient increases in external K+, which can induce voltage-dependent Cl- entry, restored GABA-induced [Ca2+]i elevations. In mature neurons, GABA reduced [Ca2+]i provided that background [Ca2+]i was elevated by the application of an L-type Ca2+ channel agonist. This was probably due to a hyperpolarization of the membrane by Cl- currents. K(+)-Cl- cotransport maintained the gradient for hyperpolarizing Cl-currents. We conclude that in immature midbrain neurons an inward Cl- transport is not effective although the GABA response is depolarizing. Further, GABA itself is not required for the developmental switch of GABAergic responses from depolarizing to hyperpolarizing in cultured midbrain neurons.

  3. Towards Real-time Metabolic Profiling of Cancer with Hyperpolarized Succinate

    PubMed Central

    Zacharias, Niki M.; McCullough, Christopher R.; Wagner, Shawn; Sailasuta, Napapon; Chan, Henry R.; Lee, Youngbok; Hu, Jingzhe; Perman, William H.; Henneberg, Cameron; Ross, Brian D.; Bhattacharya, Pratip

    2016-01-01

    Purpose The energy-yielding mitochondrial Krebs cycle has been shown in many cancers and other diseases to be inhibited or mutated. In most cells, the Krebs cycle with oxidative phosphorylation generates approximately 90% of the adenosine triphosphate in the cell. We designed and hyperpolarized carbon-13 labeled succinate (SUC) and its derivative diethyl succinate (DES) to interrogate the Krebs cycle in real-time in cancer animal models. Procedures Using Parahydrogen Induced Polarization (PHIP), we generated hyperpolarized SUC and DES by hydrogenating their respective fumarate precursors. DES and SUC metabolism was studied in five cancer allograft animal models: breast (4T1), Renal Cell Carcinoma (RENCA), colon (CT26), lymphoma NSO, and lymphoma A20. Results The extent of hyperpolarization was 8 ± 2% for SUC and 2.1 ± 0.6% for DES. The metabolism of DES and SUC in the Krebs cycle could be followed in animals 5 s after tail vein injection. The biodistribution of the compounds was observed using 13C FISP imaging. We observed significant differences in uptake and conversion of both compounds in different cell types both in vivo and in vitro. Conclusion With hyperpolarized DES and SUC, we are able to meet many of the requirements for a useable in vivo metabolic imaging compound – high polarization, relatively long T1 values, low toxicity and high water solubility. However, succinate and its derivative DES are metabolized robustly by RENCA but not by the other cancer models. Our results underscore the heterogeneity of cancer cells and the role cellular uptake plays in hyperpolarized metabolic spectroscopy. PMID:27547490

  4. In vivo single-shot 13C spectroscopic imaging of hyperpolarized metabolites by spatiotemporal encoding

    NASA Astrophysics Data System (ADS)

    Schmidt, Rita; Laustsen, Christoffer; Dumez, Jean-Nicolas; Kettunen, Mikko I.; Serrao, Eva M.; Marco-Rius, Irene; Brindle, Kevin M.; Ardenkjaer-Larsen, Jan Henrik; Frydman, Lucio

    2014-03-01

    Hyperpolarized metabolic imaging is a growing field that has provided a new tool for analyzing metabolism, particularly in cancer. Given the short life times of the hyperpolarized signal, fast and effective spectroscopic imaging methods compatible with dynamic metabolic characterizations are necessary. Several approaches have been customized for hyperpolarized 13C MRI, including CSI with a center-out k-space encoding, EPSI, and spectrally selective pulses in combination with spiral EPI acquisitions. Recent studies have described the potential of single-shot alternatives based on spatiotemporal encoding (SPEN) principles, to derive chemical-shift images within a sub-second period. By contrast to EPSI, SPEN does not require oscillating acquisition gradients to deliver chemical-shift information: its signal encodes both spatial as well as chemical shift information, at no extra cost in experimental complexity. SPEN MRI sequences with slice-selection and arbitrary excitation pulses can also be devised, endowing SPEN with the potential to deliver single-shot multi-slice chemical shift images, with a temporal resolution required for hyperpolarized dynamic metabolic imaging. The present work demonstrates this with initial in vivo results obtained from SPEN-based imaging of pyruvate and its metabolic products, after injection of hyperpolarized [1-13C]pyruvate. Multi-slice chemical-shift images of healthy rats were obtained at 4.7 T in the region of the kidney, and 4D (2D spatial, 1D spectral, 1D temporal) data sets were obtained at 7 T from a murine lymphoma tumor model.

  5. Hyperpolarizing Inhibition Develops without Trophic support by GABA in Cultured Rat Midbrain Neurons

    PubMed Central

    Titz, Stefan; Hans, Michael; Kelsch, Wolfgang; Lewen, Andrea; Swandulla, Dieter; Misgeld, Ulrich

    2003-01-01

    During a limited period of early neuronal development, GABA is depolarizing and elevates [Ca2+]i, which mediates the trophic action of GABA in neuronal maturation. We tested the attractive hypothesis that GABA itself promotes the developmental change of its response from depolarizing to hyperpolarizing (Ganguly et al. 2001). In cultured midbrain neurons we found that the GABA response changed from depolarizing to hyperpolarizing, although GABAA receptors had been blocked throughout development. In immature neurons prolonged exposure of the cells to nanomolar concentrations of GABA or brief repetitive applications of GABA strongly diminished the elevation of [Ca2+]i by GABA. As revealed by gramicidin perforated-patch recording, reduced [Ca2+]i responses were due to a diminished driving force for Cl−. This suggests that immature neurons do not have an efficient inward transport that can compensate the loss of cytosolic Cl− resulting from sustained GABAA receptor activation by ambient GABA. Transient increases in external K+, which can induce voltage-dependent Cl− entry, restored GABA-induced [Ca2+]i elevations. In mature neurons, GABA reduced [Ca2+]i provided that background [Ca2+]i was elevated by the application of an L-type Ca2+ channel agonist. This was probably due to a hyperpolarization of the membrane by Cl− currents. K+-Cl− cotransport maintained the gradient for hyperpolarizing Cl− currents. We conclude that in immature midbrain neurons an inward Cl− transport is not effective although the GABA response is depolarizing. Further, GABA itself is not required for the developmental switch of GABAergic responses from depolarizing to hyperpolarizing in cultured midbrain neurons. PMID:12938674

  6. An evaluation of potassium ions as endothelium-derived hyperpolarizing factor in porcine coronary arteries

    PubMed Central

    Bény, Jean-Louis; Schaad, Olivier

    2000-01-01

    In the rat hepatic artery, the endothelium-derived hyperpolarizing factor (EDHF) was identified as potassium. Potassium hyperpolarizes the smooth muscles by gating inward rectified potassium channels and by activating the sodium-potassium adenosine triphosphatase (Na+-K+ATPase). Our goal was to examine whether potassium could explain the EDHF in porcine coronary arteries. On coronary strips, the inhibition of calcium-dependent potassium channels with 100 nM apamin plus 100 μM charibdotoxin inhibited the endothelium-dependent relaxations, produced by 10 nM substance P and 300 nM bradykinin and resistant to nitro-L-arginine and indomethacin. The scavenging of potassium with 2 mM Kryptofix 2.2.2 abolished the endothelium-dependent relaxations produced by the kinins and resistant to nitro-L-arginine and indomethacin. Forty μM 18α glycyrrethinic acid or 50 μM palmitoleic acid, both uncoupling agents, did not inhibit these kinin relaxations. Therefore, EDHF does not result from an electrotonic spreading of an endothelial hyperpolarization. Barium (0.3 nM) did not inhibit the kinin relaxations resistant to nitro-L-arginine and indomethacin. Therefore, EDHF does not result from the activation of inward rectified potassium channels. Five hundred nM ouabain abolished the endothelium-dependent relaxations resistant to nitro-L-arginine and indomethacin without inhibiting the endothelium-derived NO relaxation. The perifusion of a medium supplemented with potassium depolarized and contracted a coronary strip; however, the short application of potassium hyperpolarized the smooth muscles. These results are compatible with the concept that, in porcine coronary artery, the EDHF is potassium released by the endothelial cells and that this ion hyperpolarizes and relaxes the smooth muscles by activating the Na+-K+ATPase. PMID:11053218

  7. Robust hyperpolarized (13)C metabolic imaging with selective non-excitation of pyruvate (SNEP).

    PubMed

    Chen, Way Cherng; Teo, Xing Qi; Lee, Man Ying; Radda, George K; Lee, Philip

    2015-08-01

    In vivo metabolic imaging using hyperpolarized [1-(13)C]pyruvate provides localized biochemical information and is particularly useful in detecting early disease changes, as well as monitoring disease progression and treatment response. However, a major limitation of hyperpolarized magnetization is its unrecoverable decay, due not only to T1 relaxation but also to radio-frequency (RF) excitation. RF excitation schemes used in metabolic imaging must therefore be able to utilize available hyperpolarized magnetization efficiently and robustly for the optimal detection of substrate and metabolite activities. In this work, a novel RF excitation scheme called selective non-excitation of pyruvate (SNEP) is presented. This excitation scheme involves the use of a spectral selective RF pulse to specifically exclude the excitation of [1-(13)C]pyruvate, while uniformly exciting the key metabolites of interest (namely [1-(13)C]lactate and [1-(13)C]alanine) and [1-(13)C]pyruvate-hydrate. By eliminating the loss of hyperpolarized [1-(13)C]pyruvate magnetization due to RF excitation, the signal from downstream metabolite pools is increased together with enhanced dynamic range. Simulation results, together with phantom measurements and in vivo experiments, demonstrated the improvement in signal-to-noise ratio (SNR) and the extension of the lifetime of the [1-(13)C]lactate and [1-(13)C]alanine pools when compared with conventional non-spectral selective (NS) excitation. SNEP has also been shown to perform comparably well with multi-band (MB) excitation, yet SNEP possesses distinct advantages, including ease of implementation, less stringent demands on gradient performance, increased robustness to frequency drifts and B0 inhomogeneity as well as easier quantification involving the use of [1-(13)C]pyruvate-hydrate as a proxy for the actual [1-(13)C] pyruvate signal. SNEP is therefore a promising alternative for robust hyperpolarized [1-(13)C]pyruvate metabolic imaging with high

  8. Gain Modulation of Cholinergic Neurons in the Medial Septum-Diagonal Band of Broca Through Hyperpolarization.

    PubMed

    Melonakos, Eric D; White, John A; Fernandez, Fernando R

    2016-12-01

    Hippocampal network oscillations are important for learning and memory. Theta rhythms are involved in attention, navigation, and memory encoding, whereas sharp wave-ripple complexes are involved in memory consolidation. Cholinergic neurons in the medial septum-diagonal band of Broca (MS-DB) influence both types of hippocampal oscillations, promoting theta rhythms and suppressing sharp wave-ripples. They also receive frequency-dependent hyperpolarizing feedback from hippocamposeptal connections, potentially affecting their role as neuromodulators in the septohippocampal circuit. However, little is known about how the integration properties of cholinergic MS-DB neurons change with hyperpolarization. By potentially altering firing behavior in cholinergic neurons, hyperpolarizing feedback from the hippocampal neurons may, in turn, change hippocampal network activity. To study changes in membrane integration properties in cholinergic neurons in response to hyperpolarizing inputs, we used whole-cell patch-clamp recordings targeting genetically labeled, choline acetyltransferase-positive neurons in mouse brain slices. Hyperpolarization of cholinergic MS-DB neurons resulted in a long-lasting decrease in spike firing rate and input-output gain. Additionally, voltage-clamp measures implicated a slowly inactivating, 4-AP-insensitive, outward K(+) conductance. Using a conductance-based model of cholinergic MS-DB neurons, we show that the ability of this conductance to modulate firing rate and gain depends on the expression of an experimentally verified shallow intrinsic spike frequency-voltage relationship. Together, these findings point to a means through which negative feedback from hippocampal neurons can influence the role of cholinergic MS-DB neurons. © 2016 Wiley Periodicals, Inc.

  9. Flow-type landslides magnitude evaluation: the case study of the Campania Region (Southern Italy)

    NASA Astrophysics Data System (ADS)

    Santo, Antonio; De Falco, Melania; Di Crescenzo, Giuseppe

    2015-04-01

    In the last years studies concerning the triggering and the run-out susceptibility for different kind of landslides have become more and more precise. In the most of the cases the methodological approach involve the production of detailed thematic maps (at least 1:5000 scale) which represent a very useful tool for territorial planning, especially in urbanized areas. More recently these researches were accompanied by the growth of other studies dealing with landslide magnitude evaluation (especially in terms of volume and velocity estimate). In this paper the results of a flow-type landslides magnitude evaluation are presented. The study area is located in Southern Italy and is very wide (1,500 square kilometres) including all the Campania region. In this context flow type landslides represent the most frequent instabilities as shown by the large number of victims and the huge economic damage caused in the last few centuries. These shallow landslides involve thin cohesionless, unsaturated pyroclastic soils found over steep slopes around Somma-Vesuvio and Phlegrean district, affecting a wide area where over 100 towns are located. Since the potential volume of flow-type landslides is a measure of event magnitude we propose to estimate the potential volume at the scale of slope or basin for about 90 municipalities affecting 850 hierarchized drainage basins and 900 regular slopes. An empirical approach recently proposed in literature (De Falco et al., 2012), allows to estimate the volume of the pyroclastic cover that can be displaced along the slope. The method derives from the interpretation of numerous geological and geomorphological data gathered from a vast amount of case histories on landslides in volcanic and carbonatic contexts and it is based on determining the thickness of the pyroclastic cover and the width of the detachment and erosion-transport zone. Thickness can be evaluated with a good degree of approximation since, in these landslides, the failure

  10. Novel Imaging Contrast Methods for Hyperpolarized 13 C Magnetic Resonance Imaging

    NASA Astrophysics Data System (ADS)

    Reed, Galen Durant

    Magnetic resonance imaging using hyperpolarized 13C-labeled small molecules has emerged as an extremely powerful tool for the in vivo monitoring of perfusion and metabolism. This work presents methods for improved imaging, parameter mapping, and image contrast generation for in vivo hyperpolarized 13C MRI. Angiography using hyperpolarized urea was greatly improved with a highly T2-weighted acquisition in combination with 15N labeling of the urea amide groups. This is due to the fact that the T2 of [13C]urea is strongly limited by the scalar coupling to the neighboring quadrupolar 14N. The long in vivo T2 values of [13C, 15N2]urea were utilized for sub-millimeter projection angiography using a contrast agent that could be safely injected in concentrations of 10-100 mM while still tolerated in patients with renal insufficiency. This study also presented the first method for in vivo T2 mapping of hyperpolarized 13C compounds. The in vivo T2 of urea was short in the blood and long within the kidneys. This persistent signal component was isolated to the renal filtrate, thus enabling for the first time direct detection of an imaging contrast agent undergoing glomerular filtration. While highly T2-weighted acquisitions select for molecules with short rotational correlation times, high diffusion weighting selects for those with the long translational correlation times. A specialized spin-echo EPI sequence was developed in order to generate highly diffusion-weighted hyperpolarized 13C images on a clinical MRI system operating within clinical peak- RF and gradient amplitude constraints. Low power adiabatic spin echo pulses were developed in order to generate a sufficiently large refocused bandwidth while maintaining low nominal power. This diffusion weighted acquisition gave enhanced tumor contrast-to-noise ratio when imaging [1-13C]lactate after infusion of [1-13C]pyruvate. Finally, the first in-man hyperpolarized 13C MRI clinical trial is discussed.

  11. A modelling study of locomotion-induced hyperpolarization of voltage threshold in cat lumbar motoneurones

    PubMed Central

    Dai, Yue; Jones, Kelvin E; Fedirchuk, Brent; McCrea, David A; Jordan, Larry M

    2002-01-01

    During fictive locomotion the excitability of adult cat lumbar motoneurones is increased by a reduction (a mean hyperpolarization of ≈6.0 mV) of voltage threshold (Vth) for action potential (AP) initiation that is accompanied by only small changes in AP height and width. Further examination of the experimental data in the present study confirms that Vth lowering is present to a similar degree in both the hyperpolarized and depolarized portions of the locomotor step cycle. This indicates that Vth reduction is a modulation of motoneurone membrane currents throughout the locomotor state rather than being related to the phasic synaptic input within the locomotor cycle. Potential ionic mechanisms of this locomotor-state-dependent increase in excitability were examined using three five-compartment models of the motoneurone innervating slow, fast fatigue resistant and fast fatigable muscle fibres. Passive and active membrane conductances were set to produce input resistance, rheobase, afterhyperpolarization (AHP) and membrane time constant values similar to those measured in adult cat motoneurones in non-locomoting conditions. The parameters of 10 membrane conductances were then individually altered in an attempt to replicate the hyperpolarization of Vth that occurs in decerebrate cats during fictive locomotion. The goal was to find conductance changes that could produce a greater than 3 mV hyperpolarization of Vth with only small changes in AP height (< 3 mV) and width (< 1.2 ms). Vth reduction without large changes in AP shape could be produced either by increasing fast sodium current or by reducing delayed rectifier potassium current. The most effective Vth reductions were achieved by either increasing the conductance of fast sodium channels or by hyperpolarizing the voltage dependency of their activation. These changes were particularly effective when localized to the initial segment. Reducing the conductance of delayed rectifier channels or depolarizing their

  12. Spreading dilatation in rat mesenteric arteries associated with calcium-independent endothelial cell hyperpolarization.

    PubMed

    Takano, Hiromichi; Dora, Kim A; Spitaler, Michaela M; Garland, Chris J

    2004-05-01

    Both ACh and levcromakalim evoke smooth muscle cell hyperpolarization and associated relaxation in rat mesenteric resistance arteries. We investigated if they could evoke conducted vasodilatation along isolated arteries, whether this reflected spreading hyperpolarization and the possible mechanism involved. Focal micropipette application of either ACh, to stimulate endothelial cell muscarinic receptors, or levcromakalim, to activate smooth muscle K(ATP) channels, each evoked a local dilatation (88 +/- 14%, n= 6 and 92 +/- 6% reversal of phenylephrine-induced tone, n= 11, respectively) that rapidly spread upstream (at 1.5 mm 46 +/- 19%, n= 6 and 57 +/- 13%, n= 9) to dilate the entire isolated artery. The local dilatation to ACh was associated with a rise in endothelial cell [Ca(2+)](i) (F/F(t = 0)= 1.22 +/- 0.33, n= 14) which did not spread beyond 0.5 mm (F/F(t = 0)= 1.01 +/- 0.01, n= 14), while the local dilatation to levcromakalim was not associated with any change in endothelial cell [Ca(2+)](i). In contrast, ACh and levcromakalim both stimulated local (12.7 +/- 1.2 mV, n= 10 and 13.5 +/- 4.7 mV, n= 10) and spreading (at 2 mm: 3.0 +/- 1.1 mV, n= 5 and 4.1 +/- 0.7 mV, n= 5) smooth muscle hyperpolarization. The spread of hyperpolarization could be prevented by cutting the artery, so was not due to a diffusible agent. Both the spreading dilatation and hyperpolarization were endothelium dependent. The injection of propidium iodide into either endothelial or smooth muscle cells revealed extensive dye coupling between the endothelial cells, but limited coupling between the smooth muscle cells. Some evidence for heterocellular spread of dye was also evident. Together, these data show that vasodilatation can spread over significant distances in mesenteric resistance arteries, and suggest this reflects an effective coupling between the endothelial cells to facilitate [Ca(2+)](i)-independent spread of hyperpolarization.

  13. Continuous-flow DNP polarizer for MRI applications at 1.5 T

    PubMed Central

    Denysenkov, V.; Terekhov, M.; Maeder, R.; Fischer, S.; Zangos, S.; Vogl, T.; Prisner, T. F.

    2017-01-01

    Here we describe a new hyperpolarization approach for magnetic resonance imaging applications at 1.5 T. Proton signal enhancements of more than 20 were achieved with a newly designed multimode microwave resonator situated inside the bore of the imager and used for Overhauser dynamic nuclear polarization of the water proton signal. Different from other approaches in our setup the hyperpolarization is achieved continuously by liquid water flowing through the polarizer under continuous microwave excitation. With an available flow rate of up to 1.5 ml/min, which should be high enough for DNP MR angiography applications in small animals like mice and rats. The hyperpolarized liquid cooled to physiological temperature can be routed by a mechanical switch to a quartz capillary for injection into the blood vessels of the target object. This new approach allows hyperpolarization of protons without the need of an additional magnet and avoids the losses arising from the transfer of the hyperpolarized solution between magnets. The signal-to-noise improvement of this method is demonstrated on two- and three-dimensional phantoms of blood vessels. PMID:28290535

  14. Continuous-flow DNP polarizer for MRI applications at 1.5 T.

    PubMed

    Denysenkov, V; Terekhov, M; Maeder, R; Fischer, S; Zangos, S; Vogl, T; Prisner, T F

    2017-03-14

    Here we describe a new hyperpolarization approach for magnetic resonance imaging applications at 1.5 T. Proton signal enhancements of more than 20 were achieved with a newly designed multimode microwave resonator situated inside the bore of the imager and used for Overhauser dynamic nuclear polarization of the water proton signal. Different from other approaches in our setup the hyperpolarization is achieved continuously by liquid water flowing through the polarizer under continuous microwave excitation. With an available flow rate of up to 1.5 ml/min, which should be high enough for DNP MR angiography applications in small animals like mice and rats. The hyperpolarized liquid cooled to physiological temperature can be routed by a mechanical switch to a quartz capillary for injection into the blood vessels of the target object. This new approach allows hyperpolarization of protons without the need of an additional magnet and avoids the losses arising from the transfer of the hyperpolarized solution between magnets. The signal-to-noise improvement of this method is demonstrated on two- and three-dimensional phantoms of blood vessels.

  15. Continuous-flow DNP polarizer for MRI applications at 1.5 T

    NASA Astrophysics Data System (ADS)

    Denysenkov, V.; Terekhov, M.; Maeder, R.; Fischer, S.; Zangos, S.; Vogl, T.; Prisner, T. F.

    2017-03-01

    Here we describe a new hyperpolarization approach for magnetic resonance imaging applications at 1.5 T. Proton signal enhancements of more than 20 were achieved with a newly designed multimode microwave resonator situated inside the bore of the imager and used for Overhauser dynamic nuclear polarization of the water proton signal. Different from other approaches in our setup the hyperpolarization is achieved continuously by liquid water flowing through the polarizer under continuous microwave excitation. With an available flow rate of up to 1.5 ml/min, which should be high enough for DNP MR angiography applications in small animals like mice and rats. The hyperpolarized liquid cooled to physiological temperature can be routed by a mechanical switch to a quartz capillary for injection into the blood vessels of the target object. This new approach allows hyperpolarization of protons without the need of an additional magnet and avoids the losses arising from the transfer of the hyperpolarized solution between magnets. The signal-to-noise improvement of this method is demonstrated on two- and three-dimensional phantoms of blood vessels.

  16. Density fluctuations in solar wind flow types at 1 AU: Comparison to Doppler scintillation measurements

    NASA Technical Reports Server (NTRS)

    Huddleston, D. E.; Woo, R.; Neugebauer, M.

    1995-01-01

    Density fluctuations with periods 10 minutes to 1 hour have been investigated in ISEE 3 plasma measurements of solar wind flows at l AU. Coronal hole, interstream, plasma sheet, coronal mass ejection, and interaction region flow types are considered. The ISEE 3 results support the interpretation of the large-scale variations in density fluctuations observed by Doppler scintillation measurement techniques inside 0.2 AU. The highest absolute and relative density fluctuations occur ahead of and within the plasma from coronal mass ejections, with the maximum values occurring between the associated interplanetary shocks and the driver gas. For the quasi-stationary solar wind, density and relative density fluctuations are highest around the heliospheric current sheet and lowest in the high-speed coronal flow. Superposed epoch analysis shows that the region of enhanced density fluctuations and its abrupt boundaries observed in the vicinity of the heliospheric current sheet near the Sun persists to l AU, providing further support for the filamentary nature of the extensions of coronal streamers. The results of this study confirm the advantages of using density fluctuations rather than density as a tracer of solar wind flows with differing origins at the Sun and as a detector of propagating interplanetary disturbances.

  17. Stimulus-evoked outer segment changes occur before the hyperpolarization of retinal photoreceptors

    PubMed Central

    Lu, Yiming; Wang, Benquan; Pepperberg, David R.; Yao, Xincheng

    2016-01-01

    Transient retinal phototropism (TRP) has been predominantly observed in rod photoreceptors activated by oblique visible light stimulation. Dynamic confocal microscopy and optical coherence tomography (OCT) have revealed rod outer segment (ROS) movement as the physical source of TRP. However, the physiological source of ROS movement is still not well understood. In this study, concurrent near-infrared imaging of TRP and electroretinogram (ERG) measurement of retinal electrophysiology revealed that ROS movement occurs before the onset of the ERG a-wave, which is known to reflect the hyperpolarization of retinal photoreceptors. Moreover, substitution of normal superfusing medium with low-sodium medium reversibly blocked the photoreceptor ERG a-wave, but largely preserved the stimulus-evoked ROS movements. Our experimental results and theoretical analysis indicate that early, disc-based stages of the phototransduction cascade, which occur before the hyperpolarization of retinal photoreceptors, contribute to the TRP associated ROS movement. PMID:28101399

  18. Liquid-state carbon-13 hyperpolarization generated in an MRI system for fast imaging

    PubMed Central

    Schmidt, A. B.; Berner, S.; Schimpf, W.; Müller, C.; Lickert, T.; Schwaderlapp, N.; Knecht, S.; Skinner, J. G.; Dost, A.; Rovedo, P.; Hennig, J.; von Elverfeldt, D.; Hövener, J. -B.

    2017-01-01

    Hyperpolarized (HP) tracers dramatically increase the sensitivity of magnetic resonance imaging (MRI) to monitor metabolism non-invasively and in vivo. Their production, however, requires an extra polarizing device (polarizer) whose complexity, operation and cost can exceed that of an MRI system itself. Furthermore, the lifetime of HP tracers is short and some of the enhancement is lost during transfer to the application site. Here, we present the production of HP tracers in water without an external polarizer: by Synthesis Amid the Magnet Bore, A Dramatically Enhanced Nuclear Alignment (SAMBADENA) is achieved within seconds, corresponding to a hyperpolarization of ∼20%. As transfer of the tracer is no longer required, SAMBADENA may permit a higher polarization at the time of detection at a fraction of the cost and complexity of external polarizers. This development is particularly promising in light of the recently extended portfolio of biomedically relevant para-hydrogen-tracers and may lead to new diagnostic applications. PMID:28262691

  19. NMR spectroscopy of hyperpolarized ^129Xe at high fields: Maintaining spin polarization after optical pumping.

    NASA Astrophysics Data System (ADS)

    Patton, Brian; Kuzma, Nicholas N.; Lisitza, Natalia V.; Happer, William

    2003-05-01

    Spin-polarized ^129Xe has become an invaluable tool in nuclear magnetic resonance research, with applications ranging from medical imaging to high-resolution spectroscopy. High-field NMR studies using hyperpolarized xenon as a spectroscopic probe benefit from the high signal-to-noise ratios and large chemical shifts typical of optically-pumped noble gases. The experimental sensitivity is ultimately determined by the absolute polarization of the xenon in the sample, which can be substantially decreased during purification and transfer. NMR of xenon at high fields (9.4 Tesla) will be discussed, and potential mechanisms of spin relaxation during the distillation, storage(N. N. Kuzma, B. Patton, K. Raman, and W. Happer, Phys. Rev. Lett. 88), 147602 (2002)., and delivery of hyperpolarized xenon will be analyzed.

  20. Light-evoked hyperpolarization and silencing of neurons by conjugated polymers

    NASA Astrophysics Data System (ADS)

    Feyen, Paul; Colombo, Elisabetta; Endeman, Duco; Nova, Mattia; Laudato, Lucia; Martino, Nicola; Antognazza, Maria Rosa; Lanzani, Guglielmo; Benfenati, Fabio; Ghezzi, Diego

    2016-03-01

    The ability to control and modulate the action potential firing in neurons represents a powerful tool for neuroscience research and clinical applications. While neuronal excitation has been achieved with many tools, including electrical and optical stimulation, hyperpolarization and neuronal inhibition are typically obtained through patch-clamp or optogenetic manipulations. Here we report the use of conjugated polymer films interfaced with neurons for inducing a light-mediated inhibition of their electrical activity. We show that prolonged illumination of the interface triggers a sustained hyperpolarization of the neuronal membrane that significantly reduces both spontaneous and evoked action potential firing. We demonstrate that the polymeric interface can be activated by either visible or infrared light and is capable of modulating neuronal activity in brain slices and explanted retinas. These findings prove the ability of conjugated polymers to tune neuronal firing and suggest their potential application for the in-vivo modulation of neuronal activity.

  1. Liquid-state carbon-13 hyperpolarization generated in an MRI system for fast imaging

    NASA Astrophysics Data System (ADS)

    Schmidt, A. B.; Berner, S.; Schimpf, W.; Müller, C.; Lickert, T.; Schwaderlapp, N.; Knecht, S.; Skinner, J. G.; Dost, A.; Rovedo, P.; Hennig, J.; von Elverfeldt, D.; Hövener, J.-B.

    2017-03-01

    Hyperpolarized (HP) tracers dramatically increase the sensitivity of magnetic resonance imaging (MRI) to monitor metabolism non-invasively and in vivo. Their production, however, requires an extra polarizing device (polarizer) whose complexity, operation and cost can exceed that of an MRI system itself. Furthermore, the lifetime of HP tracers is short and some of the enhancement is lost during transfer to the application site. Here, we present the production of HP tracers in water without an external polarizer: by Synthesis Amid the Magnet Bore, A Dramatically Enhanced Nuclear Alignment (SAMBADENA) is achieved within seconds, corresponding to a hyperpolarization of ~20%. As transfer of the tracer is no longer required, SAMBADENA may permit a higher polarization at the time of detection at a fraction of the cost and complexity of external polarizers. This development is particularly promising in light of the recently extended portfolio of biomedically relevant para-hydrogen-tracers and may lead to new diagnostic applications.

  2. Magnetic resonance imaging of pH in vivo using hyperpolarized 13C-labelled bicarbonate.

    PubMed

    Gallagher, Ferdia A; Kettunen, Mikko I; Day, Sam E; Hu, De-En; Ardenkjaer-Larsen, Jan Henrik; Zandt, René in 't; Jensen, Pernille R; Karlsson, Magnus; Golman, Klaes; Lerche, Mathilde H; Brindle, Kevin M

    2008-06-12

    As alterations in tissue pH underlie many pathological processes, the capability to image tissue pH in the clinic could offer new ways of detecting disease and response to treatment. Dynamic nuclear polarization is an emerging technique for substantially increasing the sensitivity of magnetic resonance imaging experiments. Here we show that tissue pH can be imaged in vivo from the ratio of the signal intensities of hyperpolarized bicarbonate (H(13)CO(3)(-)) and (13)CO(2) following intravenous injection of hyperpolarized H(13)CO(3)(-). The technique was demonstrated in a mouse tumour model, which showed that the average tumour interstitial pH was significantly lower than the surrounding tissue. Given that bicarbonate is an endogenous molecule that can be infused in relatively high concentrations into patients, we propose that this technique could be used clinically to image pathological processes that are associated with alterations in tissue pH, such as cancer, ischaemia and inflammation.

  3. Light-evoked hyperpolarization and silencing of neurons by conjugated polymers

    PubMed Central

    Feyen, Paul; Colombo, Elisabetta; Endeman, Duco; Nova, Mattia; Laudato, Lucia; Martino, Nicola; Antognazza, Maria Rosa; Lanzani, Guglielmo; Benfenati, Fabio; Ghezzi, Diego

    2016-01-01

    The ability to control and modulate the action potential firing in neurons represents a powerful tool for neuroscience research and clinical applications. While neuronal excitation has been achieved with many tools, including electrical and optical stimulation, hyperpolarization and neuronal inhibition are typically obtained through patch-clamp or optogenetic manipulations. Here we report the use of conjugated polymer films interfaced with neurons for inducing a light-mediated inhibition of their electrical activity. We show that prolonged illumination of the interface triggers a sustained hyperpolarization of the neuronal membrane that significantly reduces both spontaneous and evoked action potential firing. We demonstrate that the polymeric interface can be activated by either visible or infrared light and is capable of modulating neuronal activity in brain slices and explanted retinas. These findings prove the ability of conjugated polymers to tune neuronal firing and suggest their potential application for the in-vivo modulation of neuronal activity. PMID:26940513

  4. Detection of brown adipose tissue and thermogenic activity in mice by hyperpolarized xenon MRI

    PubMed Central

    Branca, Rosa Tamara; He, Ting; Zhang, Le; Floyd, Carlos S.; Freeman, Matthew; White, Christian; Burant, Alex

    2014-01-01

    The study of brown adipose tissue (BAT) in human weight regulation has been constrained by the lack of a noninvasive tool for measuring this tissue and its function in vivo. Existing imaging modalities are nonspecific and intrinsically insensitive to the less active, lipid-rich BAT of obese subjects, the target population for BAT studies. We demonstrate noninvasive imaging of BAT in mice by hyperpolarized xenon gas MRI. We detect a greater than 15-fold increase in xenon uptake by BAT during stimulation of BAT thermogenesis, which enables us to acquire background-free maps of the tissue in both lean and obese mouse phenotypes. We also demonstrate in vivo MR thermometry of BAT by hyperpolarized xenon gas. Finally, we use the linear temperature dependence of the chemical shift of xenon dissolved in adipose tissue to directly measure BAT temperature and to track thermogenic activity in vivo. PMID:25453088

  5. Cell uptake of a biosensor detected by hyperpolarized 129Xe NMR: the transferrin case.

    PubMed

    Boutin, Céline; Stopin, Antoine; Lenda, Fatimazohra; Brotin, Thierry; Dutasta, Jean-Pierre; Jamin, Nadège; Sanson, Alain; Boulard, Yves; Leteurtre, François; Huber, Gaspard; Bogaert-Buchmann, Aurore; Tassali, Nawal; Desvaux, Hervé; Carrière, Marie; Berthault, Patrick

    2011-07-01

    For detection of biological events in vitro, sensors using hyperpolarized (129)Xe NMR can become a powerful tool, provided the approach can bridge the gap in sensitivity. Here we propose constructs based on the non-selective grafting of cryptophane precursors on holo-transferrin. This biological system was chosen because there are many receptors on the cell surface, and endocytosis further increases this density. The study of these biosensors with K562 cell suspensions via fluorescence microscopy and (129)Xe NMR indicates a strong interaction, as well as interesting features such as the capacity of xenon to enter the cryptophane even when the biosensor is endocytosed, while keeping a high level of polarization. Despite a lack of specificity for transferrin receptors, undoubtedly due to the hydrophobic character of the cryptophane moiety that attracts the biosensor into the cell membrane, these biosensors allow the first in-cell probing of biological events using hyperpolarized xenon.

  6. Endothelium-dependent relaxation and hyperpolarization of canine coronary artery smooth muscles in relation to the electrogenic Na-K pump.

    PubMed Central

    Chen, G.; Hashitani, H.; Suzuki, H.

    1989-01-01

    1 In the smooth muscle cells of canine coronary artery, acetylcholine (ACh) produced a transient, endothelium-dependent hyperpolarization of the membrane. A similar hyperpolarization was also elicited by exposure to Krebs solution after incubation of the artery in K-free solution for 30 min. 2 A hyperpolarization of reproducible amplitude was generated when ACh was applied at intervals greater than 30 min. Repetitive application of ACh at 15 min intervals caused a successive reduction in the amplitude of hyperpolarization. 3 The reduction in the amplitude of relaxation during five successive applications of ACh at 15 min intervals was less than 10% of the first relaxation. 4 The ACh-induced hyperpolarization was blocked by atropine but not by ouabain, whereas the K-free induced hyperpolarization was blocked by ouabain. In low Na (Li-substituted) solution, ACh still induced a hyperpolarization but the K-free induced hyperpolarization was absent. 5 In coronary artery precontracted by high-K solution, ACh produced an endothelium-dependent relaxation, without membrane hyperpolarization. The associated relaxation was resistant to ouabain but sensitive to atropine. 6 It is concluded that in the canine coronary artery, the electrogenic Na-K pump does not contribute to the endothelium-dependent hyperpolarization or relaxation. The results are consistent with the release of two different inhibitory factors from the vascular endothelium. PMID:2590775

  7. DTG and (+)-3-PPP inhibit a ligand-activated hyperpolarization in mammalian neurons.

    PubMed

    Bobker, D H; Shen, K Z; Surprenant, A; Williams, J T

    1989-12-01

    The effects of three compounds with high affinity for the haloperidol-sensitive alpha-binding site were studied with intracellular recordings in the vitro neuronal preparations of the rat locus ceruleus, rat dorsal raphe and the guinea pig submucous plexus. Both (+)-3-(3-hydroxyphenyl)-N-(1-propyl)piperidine [(+)-3-PPP] and 1,3-di-o-tolylguanidine (DTG) inhibited the hyperpolarization induced by a ligand-activated potassium conductance. In the locus ceruleus, (+)-3-PPP and DTG produced a maximal 40 to 45% inhibition of the [Met5]enkephalin hyperpolarization, and had EC50 values of 6.6 and 2.2 microM, respectively. In the submucous plexus, the two compounds had a similar action on the alpha-2 adrenoceptor agonist UK14304 hyperpolarization, producing a maximal 50% inhibition with EC50 values of 140 and 32 nM, respectively. In addition, DTG inhibited the alpha-2-mediated inhibitory postsynaptic potential in both preparations. In contrast, (+)-3-PPP increased and prolonged the inhibitory postsynaptic potential. This action is qualitatively similar to the actions of cocaine on locus ceruleus and submucous plexus neurons. Haloperidol (1-10 microM) shared none of these actions. It is concluded that DTG and (+)-3-PPP are inhibitors of the opiate and alpha-2-mediated hyperpolarization at a postreceptor site, possibly the potassium channel. In addition, (+)-3-PPP, but not DTG, inhibits norepinephrine reuptake. None of these effects appear to be related to the sigma -binding site, because haloperidol acted as neither an agonist nor an antagonist.

  8. Hyperpolarization-activated cyclic nucleotide-gated channels in mouse vomeronasal sensory neurons.

    PubMed

    Dibattista, Michele; Mazzatenta, Andrea; Grassi, Francesca; Tirindelli, Roberto; Menini, Anna

    2008-08-01

    Hyperpolarization-activated currents (Ih) are present in several neurons of the central and peripheral nervous system. However, Ih in neurons of the vomeronasal organ (VNO) is not well characterized. We studied the properties of Ih in sensory neurons from acute slices of mouse VNO. In voltage-clamp studies, Ih was identified by the characteristic kinetics of activation, voltage dependence, and blockage by Cs+ or ZD-7288, two blockers of the Ih. Forskolin, an activator of adenylyl cyclase, shifted the activation curve for Ih to less negative potentials. A comparison of Ih properties in VNO neurons with those of heterologously expressed hyperpolarization-activated cyclic nucleotide-gated (HCN) channels, together with RT-PCR experiments in VNO, indicate that Ih is caused by HCN2 and/or HCN4 subunits. In current-clamp recordings, blocking Ih with ZD-7288 induced a hyperpolarization of 5.1 mV, an increase in input resistance, a decrease in the sensitivity to elicit action potentials in response to small current injections, and did not modify the frequency of action potentials elicited by a large current injection. It has been shown that in VNO neurons some pheromones induce a decrease in cAMP concentration, but the physiological role of cAMP is unknown. After application of blockers of adenylyl cyclase, we measured a hyperpolarization of 5.1 mV in 11 of 14 neurons, suggesting that basal levels of cAMP could modulate the resting potential. In conclusion, these results show that mouse VNO neurons express HCN2 and/or HCN4 subunits and that Ih contributes to setting the resting membrane potential and to increase excitability at stimulus threshold.

  9. High field hyperpolarization-EXSY experiment for fast determination of dissociation rates in SABRE complexes.

    PubMed

    Hermkens, Niels K J; Feiters, Martin C; Rutjes, Floris P J T; Wijmenga, Sybren S; Tessari, Marco

    2017-03-01

    SABRE (Signal Amplification By Reversible Exchange) is a nuclear spin hyperpolarization technique based on the reversible concurrent binding of small molecules and para-hydrogen (p-H2) to an iridium metal complex in solution. At low magnetic field, spontaneous conversion of p-H2 spin order to enhanced longitudinal magnetization of the nuclear spins of the other ligands occurs. Subsequent complex dissociation results in hyperpolarized substrate molecules in solution. The lifetime of this complex plays a crucial role in attained SABRE NMR signal enhancements. Depending on the ligands, vastly different dissociation rates have been previously measured using EXSY or selective inversion experiments. However, both these approaches are generally time-consuming due to the long recycle delays (up to 2min) necessary to reach thermal equilibrium for the nuclear spins of interest. In the cases of dilute solutions, signal averaging aggravates the problem, further extending the experimental time. Here, a new approach is proposed based on coherent hyperpolarization transfer to substrate protons in asymmetric complexes at high magnetic field. We have previously shown that such asymmetric complexes are important for application of SABRE to dilute substrates. Our results demonstrate that a series of high sensitivity EXSY spectra can be collected in a short experimental time thanks to the NMR signal enhancement and much shorter recycle delay.

  10. Room-temperature in situ nuclear spin hyperpolarization from optically pumped nitrogen vacancy centres in diamond

    SciTech Connect

    King, Jonathan P.; Jeong, Keunhong; Vassiliou, Christophoros C.; Shin, Chang S.; Page, Ralph H.; Avalos, Claudia E.; Wang, Hai-Jing; Pines, Alexander

    2015-12-07

    Low detection sensitivity stemming from the weak polarization of nuclear spins is a primary limitation of magnetic resonance spectroscopy and imaging. Methods have been developed to enhance nuclear spin polarization but they typically require high magnetic fields, cryogenic temperatures or sample transfer between magnets. Here we report bulk, room-temperature hyperpolarization of 13C nuclear spins observed via high-field magnetic resonance. The technique harnesses the high optically induced spin polarization of diamond nitrogen vacancy centres at room temperature in combination with dynamic nuclear polarization. We observe bulk nuclear spin polarization of 6%, an enhancement of ~170,000 over thermal equilibrium. The signal of the hyperpolarized spins was detected in situ with a standard nuclear magnetic resonance probe without the need for sample shuttling or precise crystal orientation. In conclusion, hyperpolarization via optical pumping/dynamic nuclear polarization should function at arbitrary magnetic fields enabling orders of magnitude sensitivity enhancement for nuclear magnetic resonance of solids and liquids under ambient conditions.

  11. Niflumic acid reduces the hyperpolarization-activated current (I(h)) in rod photoreceptor cells.

    PubMed

    Satoh, T O; Yamada, M

    2001-08-01

    We examined the effects of niflumic acid (NFA), a chloride channel blocker, on the hyperpolarization-activated current (I(h)) in newt rod photoreceptors. At 100 microM, NFA delayed the activation of I(h) induced by hyperpolarizing voltage pulses to -83 mV from a holding potential of -43 mV, and reduced the steady-state current. However, reduction by NFA was weakened when I(h) was activated by hyperpolarizing steps to -123 mV, suggesting that these effects were voltage-dependent. The suppressive effects of NFA on I(h) were accompanied by a negative shift in activation voltage. NFA also delayed the relaxation of I(h) tail currents, showing that this drug also inhibited deactivation of the current. The reversal potential and the fully activated conductance were not affected. These observations suggest that NFA reduces I(h) by modifying the gating kinetics of the underlying channels. The suppressive actions of NFA remained when intracellular Ca2+ was strongly chelated, and the failure of suppression by NFA in inside-out patches suggests that the agent may act on the I(h) channel from the extracellular side. These results, obtained in rod photoreceptors, are consistent with similar effects of NFA on I(f) in cardiac myocytes, suggesting that both currents share similar pharmacological properties.

  12. Room-temperature in situ nuclear spin hyperpolarization from optically pumped nitrogen vacancy centres in diamond

    DOE PAGES

    King, Jonathan P.; Jeong, Keunhong; Vassiliou, Christophoros C.; ...

    2015-12-07

    Low detection sensitivity stemming from the weak polarization of nuclear spins is a primary limitation of magnetic resonance spectroscopy and imaging. Methods have been developed to enhance nuclear spin polarization but they typically require high magnetic fields, cryogenic temperatures or sample transfer between magnets. Here we report bulk, room-temperature hyperpolarization of 13C nuclear spins observed via high-field magnetic resonance. The technique harnesses the high optically induced spin polarization of diamond nitrogen vacancy centres at room temperature in combination with dynamic nuclear polarization. We observe bulk nuclear spin polarization of 6%, an enhancement of ~170,000 over thermal equilibrium. The signal ofmore » the hyperpolarized spins was detected in situ with a standard nuclear magnetic resonance probe without the need for sample shuttling or precise crystal orientation. In conclusion, hyperpolarization via optical pumping/dynamic nuclear polarization should function at arbitrary magnetic fields enabling orders of magnitude sensitivity enhancement for nuclear magnetic resonance of solids and liquids under ambient conditions.« less

  13. Hyperpolarized 13C dehydroascorbate as an endogenous redox sensor for in vivo metabolic imaging.

    PubMed

    Keshari, Kayvan R; Kurhanewicz, John; Bok, Robert; Larson, Peder E Z; Vigneron, Daniel B; Wilson, David M

    2011-11-15

    Reduction and oxidation (redox) chemistry is involved in both normal and abnormal cellular function, in processes as diverse as circadian rhythms and neurotransmission. Intracellular redox is maintained by coupled reactions involving NADPH, glutathione (GSH), and vitamin C, as well as their corresponding oxidized counterparts. In addition to functioning as enzyme cofactors, these reducing agents have a critical role in dealing with reactive oxygen species (ROS), the toxic products of oxidative metabolism seen as culprits in aging, neurodegenerative disease, and ischemia/ reperfusion injury. Despite this strong relationship between redox and human disease, methods to interrogate a redox pair in vivo are limited. Here we report the development of [1-(13)C] dehydroascorbate [DHA], the oxidized form of Vitamin C, as an endogenous redox sensor for in vivo imaging using hyperpolarized (13)C spectroscopy. In murine models, hyperpolarized [1-(13)C] DHA was rapidly converted to [1-(13)C] vitamin C within the liver, kidneys, and brain, as well as within tumor in a transgenic prostate cancer mouse. This result is consistent with what has been previously described for the DHA/Vitamin C redox pair, and points to a role for hyperpolarized [1-(13)C] DHA in characterizing the concentrations of key intracellular reducing agents, including GSH. More broadly, these findings suggest a prognostic role for this new redox sensor in determining vulnerability of both normal and abnormal tissues to ROS.

  14. High field hyperpolarization-EXSY experiment for fast determination of dissociation rates in SABRE complexes

    NASA Astrophysics Data System (ADS)

    Hermkens, Niels K. J.; Feiters, Martin C.; Rutjes, Floris P. J. T.; Wijmenga, Sybren S.; Tessari, Marco

    2017-03-01

    SABRE (Signal Amplification By Reversible Exchange) is a nuclear spin hyperpolarization technique based on the reversible concurrent binding of small molecules and para-hydrogen (p-H2) to an iridium metal complex in solution. At low magnetic field, spontaneous conversion of p-H2 spin order to enhanced longitudinal magnetization of the nuclear spins of the other ligands occurs. Subsequent complex dissociation results in hyperpolarized substrate molecules in solution. The lifetime of this complex plays a crucial role in attained SABRE NMR signal enhancements. Depending on the ligands, vastly different dissociation rates have been previously measured using EXSY or selective inversion experiments. However, both these approaches are generally time-consuming due to the long recycle delays (up to 2 min) necessary to reach thermal equilibrium for the nuclear spins of interest. In the cases of dilute solutions, signal averaging aggravates the problem, further extending the experimental time. Here, a new approach is proposed based on coherent hyperpolarization transfer to substrate protons in asymmetric complexes at high magnetic field. We have previously shown that such asymmetric complexes are important for application of SABRE to dilute substrates. Our results demonstrate that a series of high sensitivity EXSY spectra can be collected in a short experimental time thanks to the NMR signal enhancement and much shorter recycle delay.

  15. Evaluation of hyperpolarized [1-¹³C]-pyruvate by magnetic resonance to detect ionizing radiation effects in real time.

    PubMed

    Sandulache, Vlad C; Chen, Yunyun; Lee, Jaehyuk; Rubinstein, Ashley; Ramirez, Marc S; Skinner, Heath D; Walker, Christopher M; Williams, Michelle D; Tailor, Ramesh; Court, Laurence E; Bankson, James A; Lai, Stephen Y

    2014-01-01

    Ionizing radiation (IR) cytotoxicity is primarily mediated through reactive oxygen species (ROS). Since tumor cells neutralize ROS by utilizing reducing equivalents, we hypothesized that measurements of reducing potential using real-time hyperpolarized (HP) magnetic resonance spectroscopy (MRS) and spectroscopic imaging (MRSI) can serve as a surrogate marker of IR induced ROS. This hypothesis was tested in a pre-clinical model of anaplastic thyroid carcinoma (ATC), an aggressive head and neck malignancy. Human ATC cell lines were utilized to test IR effects on ROS and reducing potential in vitro and [1-¹³C] pyruvate HP-MRS/MRSI imaging of ATC orthotopic xenografts was used to study in vivo effects of IR. IR increased ATC intra-cellular ROS levels resulting in a corresponding decrease in reducing equivalent levels. Exogenous manipulation of cellular ROS and reducing equivalent levels altered ATC radiosensitivity in a predictable manner. Irradiation of ATC xenografts resulted in an acute drop in reducing potential measured using HP-MRS, reflecting the shunting of reducing equivalents towards ROS neutralization. Residual tumor tissue post irradiation demonstrated heterogeneous viability. We have adapted HP-MRS/MRSI to non-invasively measure IR mediated changes in tumor reducing potential in real time. Continued development of this technology could facilitate the development of an adaptive clinical algorithm based on real-time adjustments in IR dose and dose mapping.

  16. Investigation of Lung Structure-Function Relationships Using Hyperpolarized Noble Gases

    NASA Astrophysics Data System (ADS)

    Thomen, Robert P.

    Magnetic Resonance Imaging (MRI) is an application of the nuclear magnetic resonance (NMR) phenomenon to non-invasively generate 3D tomographic images. MRI is an emerging modality for the lung, but it suffers from low sensitivity due to inherent low tissue density and short T(*/2) . Hyperpolarization is a process by which the nuclear contribution to NMR signal is greatly enhanced to more than 100,000 times that of samples in thermal equilibrium. The noble gases 3He and 129Xe are most often hyperpolarized by transfer of light angular momentum through the electron of a vaporized alkali metal to the noble gas nucleus (called Spin Exchange Optical Pumping). The enhancement in NMR signal is so great that the gas itself can be imaged via MRI, and because noble gases are chemically inert, they can be safely inhaled by a subject, and the gas distribution within the interior of the lung can be imaged. The mechanics of respiration is an elegant physical process by which air is is brought into the distal airspaces of the lungs for oxygen/carbon dioxide gas exchange with blood. Therefore proper description of lung function is intricately related to its physical structure , and the basic mechanical operation of healthy lungs -- from pressure driven airflow, to alveolar airspace gas kinetics, to gas exchange by blood/gas concentration gradients, to elastic contraction of parenchymal tissue -- is a process decidedly governed by the laws of physics. This dissertation will describe experiments investigating the relationship of lung structure and function using hyperpolarized (HP) noble gas MRI. In particular HP gases will be applied to the study of several pulmonary diseases each of which demonstrates unique structure-function abnormalities: asthma, cystic fibrosis, and chronic obstructive pulmonary disease. Successful implementation of an HP gas acquisition protocol for pulmonary studies is an involved and stratified undertaking which requires a solid theoretical foundation in NMR

  17. Design of a hyperpolarized (15)N NMR probe that induces a large chemical-shift change upon binding of calcium ions.

    PubMed

    Hata, Ryunosuke; Nonaka, Hiroshi; Takakusagi, Yoichi; Ichikawa, Kazuhiro; Sando, Shinsuke

    2015-08-07

    Ca(2+) is a fundamental metal ion for physiological functioning. Therefore, molecular probes for Ca(2+) analysis are required. Recently, a hyperpolarized NMR probe has emerged as a promising tool. Here, we report a new design of a hyperpolarized NMR probe for Ca(2+), which showed a large chemical shift change upon binding to Ca(2+) and was applied for Ca(2+) sensing in a hyperpolarized state.

  18. Hyperpolarized [1-13C] glutamate: a metabolic imaging biomarker of IDH1 mutational status in glioma

    PubMed Central

    Chaumeil, Myriam M.; Larson, Peder E.Z.; Woods, Sarah M.; Cai, Larry; Eriksson, Pia; Robinson, Aaron E.; Lupo, Janine M.; Vigneron, Daniel B.; Nelson, Sarah J.; Pieper, Russell O.; Phillips, Joanna J.; Ronen, Sabrina M.

    2014-01-01

    Mutations of the isocitrate dehydrogenase 1 (IDH1) gene are among the most prevalent in low-grade glioma and secondary glioblastoma, represent an early pathogenic event, and are associated with epigenetically-driven modulations of metabolism. Of particular interest is the recently uncovered relationship between the IDH1 mutation and decreased activity of the branched-chain amino acid transaminase 1 (BCAT1) enzyme. Non-invasive imaging methods that can assess BCAT1 activity could therefore improve detection of mutant IDH1 tumors and aid in developing and monitoring new targeted therapies. BCAT1 catalyzes the transamination of branched-chain amino acids while converting α-ketoglutarate (α-KG) to glutamate. Our goal was to use 13C magnetic resonance spectroscopy to probe the conversion of hyperpolarized [1-13C] α-KG to hyperpolarized [1-13C] glutamate as a readout of BCAT1 activity. We investigated two isogenic glioblastoma lines that differed only in their IDH1 status, and performed experiments in live cells and in vivo in rat orthotopic tumors. Following injection of hyperpolarized [1-13C] α-KG, hyperpolarized [1-13C] glutamate production was detected both in cells and in vivo, and the level of hyperpolarized [1-13C] glutamate was significantly lower in mutant IDH1 cells and tumors compared to their IDH1-wild-type counterparts. Importantly however, in our cells the observed drop in hyperpolarized [1-13C] glutamate was likely mediated not only by a drop in BCAT1 activity, but also by reductions in aspartate transaminase and glutamate dehydrogenase activities, suggesting additional metabolic reprogramming at least in our model. Hyperpolarized [1-13C] glutamate could thus inform on multiple mutant IDH1-associated metabolic events that mediate reduced glutamate production. PMID:24876103

  19. SUMOylation of the Hyperpolarization-Activated Cyclic Nucleotide-Gated Channel 2 Increases Surface Expression and the Maximal Conductance of the Hyperpolarization-Activated Current

    PubMed Central

    Parker, Anna R.; Welch, Meghyn A.; Forster, Lori A.; Tasneem, Sarah M.; Dubhashi, Janhavi A.; Baro, Deborah J.

    2017-01-01

    Small Ubiquitin-like Modifier (SUMO) is a ∼10 kDa peptide that can be post-translationally added to a lysine (K) on a target protein to facilitate protein–protein interactions. Recent studies have found that SUMOylation can be regulated in an activity-dependent manner and that ion channel SUMOylation can alter the biophysical properties and surface expression of the channel. Hyperpolarization-activated cyclic nucleotide-gated (HCN) channel surface expression can be regulated in an activity-dependent manner through unknown processes. We hypothesized that SUMOylation might influence the surface expression of HCN2 channels. In this manuscript, we show that HCN2 channels are SUMOylated in the mouse brain. Baseline levels of SUMOylation were also observed for a GFP-tagged HCN2 channel stably expressed in Human embryonic kidney (Hek) cells. Elevating GFP-HCN2 channel SUMOylation above baseline in Hek cells led to an increase in surface expression that augmented the hyperpolarization-activated current (Ih) mediated by these channels. Increased SUMOylation did not alter Ih voltage-dependence or kinetics of activation. There are five predicted intracellular SUMOylation sites on HCN2. Site-directed mutagenesis indicated that more than one K on the GFP-HCN2 channel was SUMOylated. Enhancing SUMOylation at one of the five predicted sites, K669, led to the increase in surface expression and Ih Gmax. The role of SUMOylation at additional sites is currently unknown. The SUMOylation site at K669 is also conserved in HCN1 channels. Aberrant SUMOylation has been linked to neurological diseases that also display alterations in HCN1 and HCN2 channel expression, such as seizures and Parkinson’s disease. This work is the first report that HCN channels can be SUMOylated and that this can regulate surface expression and Ih. PMID:28127275

  20. Demonstration of Heterogeneous Parahydrogen Induced Polarization Using Hyperpolarized Agent Migration from Dissolved Rh(I) Complex to Gas Phase

    PubMed Central

    2015-01-01

    Parahydrogen-induced polarization (PHIP) was used to demonstrate the concept that highly polarized, catalyst-free fluids can be obtained in a catalysis-free regime using a chemical reaction with molecular addition of parahydrogen to a water-soluble Rh(I) complex carrying a payload of compound with unsaturated (C=C) bonds. Hydrogenation of norbornadiene leads to formation of norbornene, which is eliminated from the Rh(I) complex and, therefore, leaves the aqueous phase and becomes a gaseous hyperpolarized molecule. The Rh(I) metal complex resides in the original liquid phase, while the product of hydrogen addition is found exclusively in the gaseous phase based on the affinity. Hyperpolarized norbornene 1H NMR signals observed in situ were enhanced by a factor of approximately 10 000 at a static field of 47.5 mT. High-resolution 1H NMR at a field of 9.4 T was used for ex situ detection of hyperpolarized norbornene in the gaseous phase, where a signal enhancement factor of approximately 160 was observed. This concept of stoichiometric as opposed to purely catalytic use of PHIP-available complexes with an unsaturated payload precursor molecule can be extended to other contrast agents for both homogeneous and heterogeneous PHIP. The Rh(I) complex was employed in aqueous medium suitable for production of hyperpolarized contrast agents for biomedical use. Detection of PHIP hyperpolarized gas by low-field NMR is demonstrated here for the first time. PMID:24918975

  1. Unmyelinated axons in the rat hippocampus hyperpolarize and activate an H current when spike frequency exceeds 1 Hz

    PubMed Central

    Soleng, A F; Chiu, K; Raastad, M

    2003-01-01

    The mammalian cortex is densely populated by extensively branching, thin, unmyelinated axons that form en passant synapses. Some thin axons in the peripheral nervous system hyperpolarize if action potential frequency exceeds 1-5 Hz. To test the hypothesis that cortical axons also show activity-induced hyperpolarization, we recorded extracellularly from individual CA3 pyramidal neurons while activating their axon with trains consisting of 30 electrical stimuli. Synaptic excitation was blocked by kynurenic acid. We observed a positive correlation between stimulation strength and the number of consecutive axonal stimuli that resulted in soma spikes, suggesting that the threshold increased as a function of the number of spikes. During trains without response failures there was always a cumulative increase in the soma response latency. Intermittent failures, however, decreased the latency of the subsequent response. At frequencies of > 1 Hz, the threshold and latency increases were enhanced by blocking the hyperpolarization-activated H current (Ih)by applying the specific Ih blocker ZD7288 (25 μM) or 2 mM Cs+. Under these conditions, response failures occurred after 15–25 stimuli, independent of the stimulation strength. Adding GABA receptor blockers (saclofen and bicuculline) and a blocker of metabotropic glutamate receptors did not change the activity-induced latency increase in recordings of the compound action potential. We interpret these results as an activity-induced hyperpolarization that is partly counteracted by Ih. Such a hyperpolarization may influence transmitter release and the conduction reliability of these axons. PMID:14561829

  2. Demonstration of heterogeneous parahydrogen induced polarization using hyperpolarized agent migration from dissolved Rh(I) complex to gas phase.

    PubMed

    Kovtunov, Kirill V; Barskiy, Danila A; Shchepin, Roman V; Coffey, Aaron M; Waddell, Kevin W; Koptyug, Igor V; Chekmenev, Eduard Y

    2014-07-01

    Parahydrogen-induced polarization (PHIP) was used to demonstrate the concept that highly polarized, catalyst-free fluids can be obtained in a catalysis-free regime using a chemical reaction with molecular addition of parahydrogen to a water-soluble Rh(I) complex carrying a payload of compound with unsaturated (C═C) bonds. Hydrogenation of norbornadiene leads to formation of norbornene, which is eliminated from the Rh(I) complex and, therefore, leaves the aqueous phase and becomes a gaseous hyperpolarized molecule. The Rh(I) metal complex resides in the original liquid phase, while the product of hydrogen addition is found exclusively in the gaseous phase based on the affinity. Hyperpolarized norbornene (1)H NMR signals observed in situ were enhanced by a factor of approximately 10,000 at a static field of 47.5 mT. High-resolution (1)H NMR at a field of 9.4 T was used for ex situ detection of hyperpolarized norbornene in the gaseous phase, where a signal enhancement factor of approximately 160 was observed. This concept of stoichiometric as opposed to purely catalytic use of PHIP-available complexes with an unsaturated payload precursor molecule can be extended to other contrast agents for both homogeneous and heterogeneous PHIP. The Rh(I) complex was employed in aqueous medium suitable for production of hyperpolarized contrast agents for biomedical use. Detection of PHIP hyperpolarized gas by low-field NMR is demonstrated here for the first time.

  3. A Large Volume Double Channel 1H-X RF Probe for Hyperpolarized Magnetic Resonance at 0.0475 Tesla

    PubMed Central

    Coffey, Aaron M.; Shchepin, Roman V.; Wilkens, Ken; Waddell, Kevin W.; Chekmenev, Eduard Y.

    2012-01-01

    In this work we describe a large volume 340 mL 1H-X magnetic resonance (MR) probe for studies of hyperpolarized compounds at 0.0475 T. 1H/13C and 1H/15N probe configurations are demonstrated with the potential for extension to 1H/129Xe. The primary applications of this probe are preparation and quality assurance of 13C and 15N hyperpolarized contrast agents using PASADENA (parahydrogen and synthesis allow dramatically enhanced nuclear alignment) and other parahydrogen-based methods of hyperpolarization. The probe is efficient and permits 62 μs 13C excitation pulses at 5.3 Watts, making it suitable for portable operation. The sensitivity and detection limits of this probe, tuned to 13C, are compared with a commercial radio frequency (RF) coil operating at 4.7 T. We demonstrate that low field MR of hyperpolarized contrast agents could be as sensitive as conventional high field detection and outline potential improvements and optimization of the probe design for preclinical in vivo MRI. PASADENA application of this low-power probe is exemplified with 13C hyperpolarized 2-hydroxyethyl propionate-1-13C,2,3,3-d3. PMID:22706029

  4. Detection of inflammatory cell function using 13C magnetic resonance spectroscopy of hyperpolarized [6-13C]-arginine

    PubMed Central

    Najac, Chloé; Chaumeil, Myriam M.; Kohanbash, Gary; Guglielmetti, Caroline; Gordon, Jeremy W.; Okada, Hideho; Ronen, Sabrina M.

    2016-01-01

    Myeloid-derived suppressor cells (MDSCs) are highly prevalent inflammatory cells that play a key role in tumor development and are considered therapeutic targets. MDSCs promote tumor growth by blocking T-cell-mediated anti-tumoral immune response through depletion of arginine that is essential for T-cell proliferation. To deplete arginine, MDSCs express high levels of arginase, which catalyzes the breakdown of arginine into urea and ornithine. Here, we developed a new hyperpolarized 13C probe, [6-13C]-arginine, to image arginase activity. We show that [6-13C]-arginine can be hyperpolarized, and hyperpolarized [13C]-urea production from [6-13C]-arginine is linearly correlated with arginase concentration in vitro. Furthermore we show that we can detect a statistically significant increase in hyperpolarized [13C]-urea production in MDSCs when compared to control bone marrow cells. This increase was associated with an increase in intracellular arginase concentration detected using a spectrophotometric assay. Hyperpolarized [6-13C]-arginine could therefore serve to image tumoral MDSC function and more broadly M2-like macrophages. PMID:27507680

  5. Crizotinib Inhibits Hyperpolarization-activated Cyclic Nucleotide-Gated Channel 4 Activity

    PubMed Central

    Zhang, Zhushan; Huang, Tai-Qin; Nepliouev, Igor; Zhang, Hengtao; Barnett, Adam S.; Rosenberg, Paul B.; Ou, Sai-Hong I.; Stiber, Jonathan A.

    2017-01-01

    Background Sinus bradycardia is frequently observed in patients treated with crizotinib, a receptor tyrosine kinase inhibitor used for the treatment of anaplastic lymphoma kinase (ALK)-rearranged non-small cell lung cancer (NSCLC). We investigated whether crizotinib could influence heart rate (HR) through direct cardiac effects. Methods The direct effect of crizotinib on HR was studied using ECG analysis of Langendorff-perfused mouse hearts. The whole-cell patch clamp technique was used to measure the effects of crizotinib on the hyperpolarization-activated funny current, If, in mouse sinoatrial node cells (SANCs) and hyperpolarization-activated cyclic nucleotide-gated channel 4 (HCN4) activity in HEK-293 cells stably expressing human HCN4. Results Crizotinib resulted in a dose-dependent reduction in HR in isolated intact mouse hearts with a half maximal inhibitory concentration (IC50) of 1.7 ± 0.4 μmol/L. Because ECG analysis revealed that crizotinib (0–5 μmol/L) resulted in significant reductions in HR in isolated mouse hearts without changes in PR, QRS, or QT intervals, we performed whole-cell patch clamp recordings of SANCs which showed that crizotinib inhibited If which regulates cardiac pacemaker activity. Crizotinib resulted in diminished current density of HCN4, the major molecular determinant of If, with an IC50 of 1.4 ± 0.3 μmol/L. Crizotinib also slowed HCN4 activation and shifted the activation curve to the left towards more hyperpolarized potentials. Conclusions Our results suggest that crizotinib’s effects on HCN4 channels play a significant role in mediating its observed effects on HR. PMID:28217366

  6. Inhibition of the hyperpolarization-activated current (if) of rabbit SA node myocytes by niflumic acid.

    PubMed

    Accili, E A; DiFrancesco, D

    1996-03-01

    The effects of the amphiphilic substance niflumic acid (NFA) were examined in myocytes isolated from the sino-atrial node of the rabbit heart. NFA (50 and 500 microM), for 30-60 s, produced a reversible negative chronotropic effect by reducing the rate of diastolic depolarization, suggesting an inhibitory effect on the hyperpolarization-activated pacemaker current (if). NFA (from 0.05 to 500 microM) inhibited if by modifying the current kinetics, without alteration of the conductance. This was shown by evidence indicating that: (1) NFA inhibited if during hyperpolarizing pulses to the mid-point of if activation but not at fully activating voltages; (2) the slope and reversal potential of the fully activated current/voltage (I/V) relation were not altered by NFA, indicating no change in slope conductance or ion selectivity; and (3) hyperpolarizing ramp protocols confirmed the lack of action of 50 microM NFA on the fully activated current and a shift of approximately -8 mV. Although similar to inhibition by acetylcholine (ACh), inhibition by NFA was only partly additive with the action of ACh and was not altered by atropine or pertussis toxin, both of which eliminated the action of ACh. The effect of NFA was present after stimulation of adenylate cyclase by forskolin and after inhibition of phosphodiesterase by isobutylmethylxanthine (IBMX). In cell-attached patch measurements, NFA applied externally did not affect if measured in the patch. Finally, application of NFA to the cytoplasmic side of excised patches did not alter the current in the absence or presence of adenosine 3',5'-cyclic monophosphate (cAMP). These results suggest an external, membrane-delimited action of NFA on if.

  7. Cardiac perfusion imaging using hyperpolarized 13c urea using flow sensitizing gradients

    PubMed Central

    Miller, Jack J.; Robson, Matthew D.; Tyler, Damian J.

    2015-01-01

    Purpose To demonstrate the feasibility of imaging the first passage of a bolus of hyperpolarized 13C urea through the rodent heart using flow‐sensitizing gradients to reduce signal from the blood pool. Methods A flow‐sensitizing bipolar gradient was optimized to reduce the bright signal within the cardiac chambers, enabling improved contrast of the agent within the tissue capillary bed. The gradient was incorporated into a dynamic golden angle spiral 13C imaging sequence. Healthy rats were scanned during rest (n = 3) and under adenosine stress‐induced hyperemia (n = 3). Results A two‐fold increase in myocardial perfusion relative to rest was detected during adenosine stress‐induced hyperemia, consistent with a myocardial perfusion reserve of two in rodents. Conclusion The new pulse sequence was used to obtain dynamic images of the first passage of hyperpolarized 13C urea in the rodent heart, without contamination from bright signal within the neighboring cardiac lumen. This probe of myocardial perfusion is expected to enable new hyperpolarized 13C studies in which the cardiac metabolism/perfusion mismatch can be identified. Magn Reson Med, 2015. © 2015 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. Magn Reson Med 75:1474–1483, 2016. © 2015 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance. PMID:25991580

  8. Metabolism of hyperpolarized [1‐13C]pyruvate through alternate pathways in rat liver

    PubMed Central

    Moreno, Karlos X.; Wang, Jian‐Xiong; Fidelino, Leila; Merritt, Matthew E.; Sherry, A. Dean; Malloy, Craig R.

    2016-01-01

    The source of hyperpolarized (HP) [13C]bicarbonate in the liver during metabolism of HP [1‐13C]pyruvate is uncertain and likely changes with physiology. Multiple processes including decarboxylation through pyruvate dehydrogenase or pyruvate carboxylase followed by subsequent decarboxylation via phosphoenolpyruvate carboxykinase (gluconeogenesis) could play a role. Here we tested which metabolic fate of pyruvate contributed to the appearance of HP [13C]bicarbonate during metabolism of HP [1‐13C]pyruvate by the liver in rats after 21 h of fasting compared to rats with free access to food. The 13C NMR of HP [13C]bicarbonate was observed in the liver of fed rats, but not in fasted rats where pyruvate carboxylation and gluconeogenesis was active. To further explore the relative fluxes through pyruvate carboxylase versus pyruvate dehydrogenase in the liver under typical conditions of hyperpolarization studies, separate parallel experiments were performed with rats given non‐hyperpolarized [2,3‐13C]pyruvate. 13C NMR analysis of glutamate isolated from the liver of rats revealed that flux from injected pyruvate through pyruvate dehydrogenase was dominant under fed conditions whereas flux through pyruvate carboxylase dominated under fasted conditions. The NMR signal of HP [13C]bicarbonate does not parallel pyruvate carboxylase activity followed by subsequent decarboxylation reaction leading to glucose production. In the liver of healthy well‐fed rats, the appearance of HP [13C]bicarbonate exclusively reflects decarboxylation of HP [1‐13C]pyruvate via pyruvate dehydrogenase. © 2016 The Authors. NMR in Biomedicine published by John Wiley & Sons Ltd. PMID:26836042

  9. Metabolism of hyperpolarized [1-(13)C]pyruvate through alternate pathways in rat liver.

    PubMed

    Jin, Eunsook S; Moreno, Karlos X; Wang, Jian-Xiong; Fidelino, Leila; Merritt, Matthew E; Sherry, A Dean; Malloy, Craig R

    2016-04-01

    The source of hyperpolarized (HP) [(13)C]bicarbonate in the liver during metabolism of HP [1-(13)C]pyruvate is uncertain and likely changes with physiology. Multiple processes including decarboxylation through pyruvate dehydrogenase or pyruvate carboxylase followed by subsequent decarboxylation via phosphoenolpyruvate carboxykinase (gluconeogenesis) could play a role. Here we tested which metabolic fate of pyruvate contributed to the appearance of HP [(13)C]bicarbonate during metabolism of HP [1-(13)C]pyruvate by the liver in rats after 21 h of fasting compared to rats with free access to food. The (13)C NMR of HP [(13)C]bicarbonate was observed in the liver of fed rats, but not in fasted rats where pyruvate carboxylation and gluconeogenesis was active. To further explore the relative fluxes through pyruvate carboxylase versus pyruvate dehydrogenase in the liver under typical conditions of hyperpolarization studies, separate parallel experiments were performed with rats given non-hyperpolarized [2,3-(13)C]pyruvate. (13)C NMR analysis of glutamate isolated from the liver of rats revealed that flux from injected pyruvate through pyruvate dehydrogenase was dominant under fed conditions whereas flux through pyruvate carboxylase dominated under fasted conditions. The NMR signal of HP [(13)C]bicarbonate does not parallel pyruvate carboxylase activity followed by subsequent decarboxylation reaction leading to glucose production. In the liver of healthy well-fed rats, the appearance of HP [(13)C]bicarbonate exclusively reflects decarboxylation of HP [1-(13)C]pyruvate via pyruvate dehydrogenase.

  10. A comparison of quantitative methods for clinical imaging with hyperpolarized (13)C-pyruvate.

    PubMed

    Daniels, Charlie J; McLean, Mary A; Schulte, Rolf F; Robb, Fraser J; Gill, Andrew B; McGlashan, Nicholas; Graves, Martin J; Schwaiger, Markus; Lomas, David J; Brindle, Kevin M; Gallagher, Ferdia A

    2016-04-01

    Dissolution dynamic nuclear polarization (DNP) enables the metabolism of hyperpolarized (13)C-labelled molecules, such as the conversion of [1-(13)C]pyruvate to [1-(13)C]lactate, to be dynamically and non-invasively imaged in tissue. Imaging of this exchange reaction in animal models has been shown to detect early treatment response and correlate with tumour grade. The first human DNP study has recently been completed, and, for widespread clinical translation, simple and reliable methods are necessary to accurately probe the reaction in patients. However, there is currently no consensus on the most appropriate method to quantify this exchange reaction. In this study, an in vitro system was used to compare several kinetic models, as well as simple model-free methods. Experiments were performed using a clinical hyperpolarizer, a human 3 T MR system, and spectroscopic imaging sequences. The quantitative methods were compared in vivo by using subcutaneous breast tumours in rats to examine the effect of pyruvate inflow. The two-way kinetic model was the most accurate method for characterizing the exchange reaction in vitro, and the incorporation of a Heaviside step inflow profile was best able to describe the in vivo data. The lactate time-to-peak and the lactate-to-pyruvate area under the curve ratio were simple model-free approaches that accurately represented the full reaction, with the time-to-peak method performing indistinguishably from the best kinetic model. Finally, extracting data from a single pixel was a robust and reliable surrogate of the whole region of interest. This work has identified appropriate quantitative methods for future work in the analysis of human hyperpolarized (13)C data.

  11. Characteristics and physiological role of hyperpolarization activated currents in mouse cold thermoreceptors

    PubMed Central

    Orio, Patricio; Madrid, Rodolfo; de la Peña, Elvira; Parra, Andrés; Meseguer, Víctor; Bayliss, Douglas A; Belmonte, Carlos; Viana, Félix

    2009-01-01

    Hyperpolarization-activated currents (Ih) are mediated by the expression of combinations of hyperpolarization-activated, cyclic nucleotide-gated (HCN) channel subunits (HCN1–4). These cation currents are key regulators of cellular excitability in the heart and many neurons in the nervous system. Subunit composition determines the gating properties and cAMP sensitivity of native Ih currents. We investigated the functional properties of Ih in adult mouse cold thermoreceptor neurons from the trigeminal ganglion, identified by their high sensitivity to moderate cooling and responsiveness to menthol. All cultured cold-sensitive (CS) neurons expressed a fast activating Ih, which was fully blocked by extracellular Cs+ or ZD7288 and had biophysical properties consistent with those of heteromeric HCN1–HCN2 channels. In CS neurons from HCN1(−/−) animals, Ih was greatly reduced but not abolished. We find that Ih activity is not essential for the transduction of cold stimuli in CS neurons. Nevertheless, Ih has the potential to shape the excitability of CS neurons. First, Ih blockade caused a membrane hyperpolarization in CS neurons of about 5 mV. Furthermore, impedance power analysis showed that all CS neurons had a prominent subthreshold membrane resonance in the 5–7 Hz range, completely abolished upon blockade of Ih and absent in HCN1 null mice. This frequency range matches the spontaneous firing frequency of cold thermoreceptor terminals in vivo. Behavioural responses to cooling were reduced in HCN1 null mice and after peripheral pharmacological blockade of Ih with ZD7288, suggesting that Ih plays an important role in peripheral sensitivity to cold. PMID:19273581

  12. Nuclear spin imaging with hyperpolarized nuclei created by brute force method

    NASA Astrophysics Data System (ADS)

    Tanaka, Masayoshi; Kunimatsu, Takayuki; Fujiwara, Mamoru; Kohri, Hideki; Ohta, Takeshi; Utsuro, Masahiko; Yosoi, Masaru; Ono, Satoshi; Fukuda, Kohji; Takamatsu, Kunihiko; Ueda, Kunihiro; Didelez, Jean-P.; Prossati, Giorgio; de Waard, Arlette

    2011-05-01

    We have been developing a polarized HD target for particle physics at the SPring-8 under the leadership of the RCNP, Osaka University for the past 5 years. Nuclear polarizaton is created by means of the brute force method which uses a high magnetic field (~17 T) and a low temperature (~ 10 mK). As one of the promising applications of the brute force method to life sciences we started a new project, "NSI" (Nuclear Spin Imaging), where hyperpolarized nuclei are used for the MRI (Magnetic Resonance Imaging). The candidate nuclei with spin ½hslash are 3He, 13C, 15N, 19F, 29Si, and 31P, which are important elements for the composition of the biomolecules. Since the NMR signals from these isotopes are enhanced by orders of magnitudes, the spacial resolution in the imaging would be much more improved compared to the practical MRI used so far. Another advantage of hyperpolarized MRI is that the MRI is basically free from the radiation, while the problems of radiation exposure caused by the X-ray CT or PET (Positron Emission Tomography) cannot be neglected. In fact, the risk of cancer for Japanese due to the radiation exposure through these diagnoses is exceptionally high among the advanced countries. As the first step of the NSI project, we are developing a system to produce hyperpolarized 3He gas for the diagnosis of serious lung diseases, for example, COPD (Chronic Obstructive Pulmonary Disease). The system employs the same 3He/4He dilution refrigerator and superconducting solenoidal coil as those used for the polarized HD target with some modification allowing the 3He Pomeranchuk cooling and the following rapid melting of the polarized solid 3He to avoid the depolarization. In this report, the present and future steps of our project will be outlined with some latest experimental results.

  13. Hyperpolarized 129Xe magnetic resonance imaging of a rat model of transient Ischemic Stroke

    NASA Astrophysics Data System (ADS)

    Walvick, Ronn P.; Bastan, Birgul; Reno, Austin; Mansour, Joey; Sun, Yanping; Zhou, Xin; Mazzani, Mary; Fisher, Marc; Sotak, Christopher H.; Albert, Mitchell S.

    2009-02-01

    Ischemic stroke accounts for nearly 80% of all stroke cases. Although proton diffusion and perfusion magnetic resonance imaging (MRI) are the gold standards in ischemic stroke diagnostics, the use of hyperpolarized 129Xe MRI has a potential role to contribute to the diagnostic picture. The highly lipophilic hyperpolarized 129Xe can be non-invasively delivered via inhalation into the lungs where it is dissolved into the blood and delivered to other organs such as the brain. As such, we expect hyperpolarized 129Xe to act as a perfusion tracer which will result in a signal deficit in areas of blood deprived tissue. In this work, we present imaging results from an animal model of transient ischemic stroke characterized through 129Xe MRI. In this model, a suture is used to occlude the middle cerebral artery (MCA) in the rat brain, thus causing an ischemic event. After a period of MCA occlusion, the suture can then be removed to reperfuse the ischemic area. During the ischemic phase of the stroke, a signal void was observed in the MCA territory; which was subsequently restored by normal 129Xe MRI signal once perfusion was reinstated. Further, a higher resolution one-dimensional chemical shift image shows a sharp signal drop in the area of ischemia. Validation of ischemic damage was shown through both proton diffusion-weighted MRI (DWI) and by 2,3,5-triphenyltetrazoliumchloride (TTC) staining. The results show the potential of 129Xe to act as a perfusion tracer; information that may add to the diagnostic and prognostic utility of the clinical picture of stroke.

  14. Development and testing of hyperpolarized 13C MR calibrationless parallel imaging

    PubMed Central

    Feng, Yesu; Gordon, Jeremy W.; Shin, Peter J.; von Morze, Cornelius; Lustig, Michael; Larson, Peder E.Z.; Ohliger, Michael A.; Carvajal, Lucas; Tropp, James; Pauly, John M.; Vigneron, Daniel B.

    2016-01-01

    A calibrationless parallel imaging technique developed previously for 1H MRI was modified and tested for hyperpolarized 13C MRI for applications requiring large FOV and high spatial resolution. The technique was demonstrated with both retrospective and prospective under-sampled data acquired in phantom and in vivo rat studies. A 2-fold acceleration was achieved using a 2D symmetric EPI readout equipped with random blips on the phase encode dimension. Reconstructed images showed excellent qualitative agreement with fully sampled data. Further acceleration can be achieved using acquisition schemes that incorporate multi-dimensional under-sampling. PMID:26679288

  15. Rapid longitudinal relaxation measurement of hyperpolarized 129Xe by a highly sensitive atomic magnetometer

    NASA Astrophysics Data System (ADS)

    Li, Yingying; Wang, Zhiguo; Jin, Shilong; Yuan, Jie; Zhao, Hongchang; Luo, Hui

    2017-02-01

    A fast and accurate determination of longitudinal relaxation time is put forward for hyperpolarized 129Xe. The theoretical framework for the method is developed. Measurement of the longitudinal relaxation time is by the determination of a close-loop response of 129Xe magnetization to the external magnetic fields and is implemented with a highly sensitive Rb magnetometer. The indirect measurement dramatically reduces the time consuming than the conventional inversion-recovery method and is more suitable for the samples with long longitudinal relaxation time.

  16. Boron Induces Hyperpolarization of Sunflower Root Cell Membranes and Increases Membrane Permeability to K+1

    PubMed Central

    Schon, Mary K.; Novacky, Anton; Blevins, Dale G.

    1990-01-01

    Although many studies have alluded to a role for boron (B) in membrane function, there is little evidence for a direct effect of B on the plasmalemma of higher plant cells. These studies were conducted to demonstrate, by electrophysiological techniques, a direct effect of B on the membrane potential (Em) of sunflower (Helianthus annuus [L.], cv Mammoth Grey Stripe) root tip cells and to determine if the response to B occurs rapidly enough to account for the previously observed effects of B on ion uptake. By inserting a glass microelectrode into an individual cell in the root tip, the Em of the cell was determined in basal salt medium (BSM), pH 6.0. The perfusion solution surrounding the root tissue was then changed to BSM + 50 micromolar H3BO3, pH 6.0. The exposure to B induced a significant plasmalemma hyperpolarization in sunflower root cells within 20 minutes. After just 3 minutes of exposure to B, the change in Em was already significantly different from the negligible change in Em observed over time in root cells never exposed to B. Membrane hyperpolarization could be caused by a stimulation of the proton pump or by a change in the conductance of one or more permeable ions. Since B has been shown to affect K+ uptake by plants, the electrophysiological techniques described above were used to determine if B has an effect on membrane permeability to K+, and could thereby lead to an increased diffusion potential. When sunflower root tips were pretreated in 50 micromolar B for 2 hours, cell membranes exhibited a significantly greater depolarization with each 10-fold increase in external [K+] than minus-B cells. Subsequent studies demonstrated that the depolarization due to increased external [K+] was also significantly greater when tissue was exposed to B at the same time as the 10-fold increase in [K+], indicating that the effect of B on K+ permeability was immediate. Analysis of sunflower root tips demonstrated that treatment in 50 micromolar B caused a

  17. The Relationship between NMR Chemical Shifts of Thermally Polarized and Hyperpolarized (89) Y Complexes and Their Solution Structures.

    PubMed

    Xing, Yixun; Jindal, Ashish K; Regueiro-Figueroa, Martín; Le Fur, Mariane; Kervarec, Nelly; Zhao, Piyu; Kovacs, Zoltan; Valencia, Laura; Pérez-Lourido, Paulo; Tripier, Raphaël; Esteban-Gómez, David; Platas-Iglesias, Carlos; Sherry, A Dean

    2016-11-07

    Recently developed dynamic nuclear polarization (DNP) technology offers the potential of increasing the NMR sensitivity of even rare nuclei for biological imaging applications. Hyperpolarized (89) Y is an ideal candidate because of its narrow NMR linewidth, favorable spin quantum number (I=1/2 ), and long longitudinal relaxation times (T1 ). Strong NMR signals were detected in hyperpolarized (89) Y samples of a variety of yttrium complexes. A dataset of (89) Y NMR data composed of 23 complexes with polyaminocarboxylate ligands was obtained using hyperpolarized (89) Y measurements or (1) H,(89) Y-HMQC spectroscopy. These data were used to derive an empirical equation that describes the correlation between the (89) Y chemical shift and the chemical structure of the complexes. This empirical correlation serves as a guide for the design of (89) Y sensors. Relativistic (DKH2) DFT calculations were found to predict the experimental (89) Y chemical shifts to a rather good accuracy.

  18. In vivo detection of cucurbit[6]uril, a hyperpolarized xenon contrast agent for a xenon magnetic resonance imaging biosensor

    NASA Astrophysics Data System (ADS)

    Hane, Francis T.; Li, Tao; Smylie, Peter; Pellizzari, Raiili M.; Plata, Jennifer A.; Deboef, Brenton; Albert, Mitchell S.

    2017-01-01

    The Hyperpolarized gas Chemical Exchange Saturation Transfer (HyperCEST) Magnetic Resonance (MR) technique has the potential to increase the sensitivity of a hyperpolarized xenon-129 MRI contrast agent. Signal enhancement is accomplished by selectively depolarizing the xenon within a cage molecule which, upon exchange, reduces the signal in the dissolved phase pool. Herein we demonstrate the in vivo detection of the cucurbit[6]uril (CB6) contrast agent within the vasculature of a living rat. Our work may be used as a stepping stone towards using the HyperCEST technique as a molecular imaging modality.

  19. In vivo detection of cucurbit[6]uril, a hyperpolarized xenon contrast agent for a xenon magnetic resonance imaging biosensor

    PubMed Central

    Hane, Francis T.; Li, Tao; Smylie, Peter; Pellizzari, Raiili M.; Plata, Jennifer A.; DeBoef, Brenton; Albert, Mitchell S.

    2017-01-01

    The Hyperpolarized gas Chemical Exchange Saturation Transfer (HyperCEST) Magnetic Resonance (MR) technique has the potential to increase the sensitivity of a hyperpolarized xenon-129 MRI contrast agent. Signal enhancement is accomplished by selectively depolarizing the xenon within a cage molecule which, upon exchange, reduces the signal in the dissolved phase pool. Herein we demonstrate the in vivo detection of the cucurbit[6]uril (CB6) contrast agent within the vasculature of a living rat. Our work may be used as a stepping stone towards using the HyperCEST technique as a molecular imaging modality. PMID:28106110

  20. BI-07HYPERPOLARIZED [1-13C] GLUTAMATE: A METABOLIC IMAGING BIOMARKER OF IDH1 MUTATIONAL STATUS IN GLIOMA

    PubMed Central

    Chaumeil, Myriam; Larson, Peder; Woods, Sarah; Cai, Larry; Eriksson, Pia; Robinson, Aaron; Lupo, Janine; Vigneron, Daniel; Nelson, Sarah; Pieper, Russell; Phillips, Joanna; Ronen, Sabrina

    2014-01-01

    Mutations of the isocitrate dehydrogenase 1 (IDH1) gene are among the most prevalent in low-grade glioma and secondary glioblastoma, represent an early pathogenic event and are being considered a promising therapeutic target. Consequently, non-invasive imaging methods are needed to monitor IDH1 status. Amongst these, we previously demonstrated the use of 13C MR spectroscopic imaging of hyperpolarized [1-13C] α-ketoglutarate (α-KG) to non-invasively assess IDH1 status through the detection of the conversion of hyperpolarized α-ketoglutarate to 2-hydroxyglutarate (2-HG) catalyzed by mutant IDH1. Importantly, in addition to its oncogenic role, IDH1 mutation is also associated with global modulations in metabolism. Interestingly, a study recently uncovered a relationship between presence of IDH1 mutation and decreased activity of the branched chained amino acid transaminase 1 (BCAT1) enzyme, which transaminates amino acids while converting α-KG to glutamate. Given this new study, we decided to expand on our previous findings and investigated the potential of hyperpolarized α-KG as an imaging probe to monitor BCAT1-driven α-KG-to-glutamate conversion and its modulation in the presence of IDH1 mutation. We investigated two isogenic glioblastoma lines that differed only in their IDH1 status, and performed experiments in live cells and in vivo in rat orthotopic tumors. Following injection of hyperpolarized α-KG, hyperpolarized glutamate production was detected both in cells and in vivo, and the level of hyperpolarized glutamate was significantly lower in mutant IDH1 cells and tumors compared to their IDH1-wild-type counterparts. Importantly however, the observed drop in hyperpolarized glutamate was likely mediated not only by a drop in BCAT1 activity, but also by reductions in aspartate transaminase and glutamate dehydrogenase activities, suggesting additional metabolic reprogramming at least in our model. Hyperpolarized glutamate could thus inform on multiple

  1. Alterations in endothelium-dependent hyperpolarization and relaxation in mesenteric arteries from streptozotocin-induced diabetic rats

    PubMed Central

    Fukao, Mitsuhiro; Hattori, Yuichi; Kanno, Morio; Sakuma, Ichiro; Kitabatake, Akira

    1997-01-01

    The aim of this study was to determine whether endothelium-dependent hyperpolarization and relaxation are altered during experimental diabetes mellitus. Membrane potentials were recorded in mesenteric arteries from rats with streptozotocin-induced diabetes and age-matched controls. The resting membrane potentials were not significantly different between control and diabetic mesenteric arteries (−55.3±0.5 vs −55.6±0.4 mV). However, endothelium-dependent hyperpolarization produced by acetylcholine (ACh; 10−8–10−5 M) was significantly diminished in amplitude in diabetic arteries compared with that in controls (maximum −10.4±1.1 vs −17.2±0.8 mV). Furthermore, the hyperpolarizing responses of diabetic arteries were more transient. ACh-induced hyperpolarization observed in control and diabetic arteries remained unaltered even after treatment with 3×10−4 M NG-nitro-L-arginine (L-NOARG), 10−5 M indomethacin or 60 u ml−1 superoxide dismutase. Endothelium-dependent hyperpolarization with 10−6 M A23187, a calcium ionophore, was also decreased in diabetic arteries compared to controls (−8.3±1.4 vs −18.0±1.9 mV). However, endothelium-independent hyperpolarizing responses to 10−6 M pinacidil, a potassium channel opener, were similar in control and diabetic arteries (−20.0±1.4 vs −19.2±1.1 mV). The altered endothelium-dependent hyperpolarizations in diabetic arteries were almost completely prevented by insulin therapy. Endothelium-dependent relaxations by ACh in the presence of 10−4 M L-NOARG and 10−5 M indomethacin in diabetic arteries were also reduced and more transient compared to controls. These data indicate that endothelium-dependent hyperpolarization is reduced by diabetes, and this would, in part, account for the impaired endothelium-dependent relaxations in mesenteric arteries from diabetic rats. PMID:9257918

  2. Correlation of measures of regional lung ventilation from 4DCT vs. hyperpolarized helium-3 MR

    NASA Astrophysics Data System (ADS)

    Ding, Kai; Cao, Kunlin; Miller, Wilson; Christensen, Gary; Reinhardt, Joseph; Benedict, Stanley; Libby, Bruce; Sheng, Ke

    2012-03-01

    Radiation induced pulmonary diseases can change the tissue material properties of lung parenchyma and the mechanics of the respiratory system. Recent advances in multi-detector-row CT (MDCT), 4DCT respiratory gating methods, and image processing techniques enable us to follow and measure those changes noninvasively during radiation therapy at a regional level. This study compares the 4DCT based ventilation measurement with the results from hyperpolarized helium-3 MR using the cumulative distribution function maps and the relative overlap (RO) statistic. We show that the similarity between the two measurements increases as the increase of the B-Spline grid spacing and Laplacian weighting which result a smoother ventilation map. The best similarity is found with weighting of 0.5 for linear elasticity and B-Spline grid spacing of 32 mm. Future work is to improve the lung image registration algorithm by incorporating hyperpolarized helium-3 MR information so as to improve its physiological modeling of the lung tissue deformation.

  3. Spin Noise Detection of Nuclear Hyperpolarization at 1.2 K

    PubMed Central

    Pöschko, Maria Theresia; Vuichoud, Basile; Milani, Jonas; Bornet, Aurélien; Bechmann, Matthias; Bodenhausen, Geoffrey; Jannin, Sami; Müller, Norbert

    2015-01-01

    We report proton spin noise spectra of a hyperpolarized solid sample of commonly used “DNP (dynamic nuclear polarization) juice” containing TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidine N-oxide) and irradiated by a microwave field at a temperature of 1.2 K in a magnetic field of 6.7 T. The line shapes of the spin noise power spectra are sensitive to the variation of the microwave irradiation frequency and change from dip to bump, when the electron Larmor frequency is crossed, which is shown to be in good accordance with theory by simulations. Small but significant deviations from these predictions are observed, which can be related to spin noise and radiation damping phenomena that have been reported in thermally polarized systems. The non-linear dependence of the spin noise integral on nuclear polarization provides a means to monitor hyperpolarization semi-quantitatively without any perturbation of the spin system by radio frequency irradiation. PMID:26477605

  4. Large dose hyperpolarized water with dissolution-DNP at high magnetic field

    NASA Astrophysics Data System (ADS)

    Lipsø, Kasper Wigh; Bowen, Sean; Rybalko, Oleksandr; Ardenkjær-Larsen, Jan Henrik

    2017-01-01

    We demonstrate a method for the preparation of hyperpolarized water by dissolution Dynamic Nuclear Polarization at high magnetic field. Protons were polarized at 6.7 T and 1.1 K to >70% with frequency modulated microwave irradiation at 188G Hz. 97.2 ± 0.7% of the radical was extracted from the sample in the dissolution in a two-phase system. 16 ± 1 mL of 5.0 M 1H in D2O with a polarization of 13.0 ± 0.9% in the liquid state was obtained, corresponding to an enhancement factor of 4000 ± 300 compared to the thermal equilibrium at 9.4 T and 293 K. A longitudinal relaxation time constant of 16 ± 1 s was measured. The sample was polarized and dissolved in a fluid path compatible with clinical polarizers. The volume of hyperpolarized water produced by this method enables angiography and perfusion measurements in large animals, as well as NMR experiments for studies of e.g. proton exchange and polarization transfer to other nuclei.

  5. Effects of N-glycosylation on hyperpolarization-activated cyclic nucleotide-gated (HCN) channels.

    PubMed

    Li, Mo; Tonggu, Lige; Tang, Lan; Wang, Liguo

    2015-02-15

    Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are activated by membrane hyperpolarization and conduct an inward cation current, which contributes to rhythmic electrical activity of neural and cardiac pacemaker cells. HCN channels have been shown to undergo N-linked glycosylation, and the N-glycosylation has been shown to be required for membrane trafficking and possibly function. In this study, recombinant wild-type (WT) and glycosylation-defective N380Q HCN2 channels were individually or co-expressed in HEK-293 cells. We demonstrate that glycosylation is required for trafficking to the plasma membrane and for the stability of HCN channels in the cell. Interestingly, the heteromeric HCN2 channels of WT and glycosylation-defective N380Q have been observed on cell membranes, indicating that not all four subunits of a tetrameric HCN2 channel need to be glycosylated for HCN2 channels to traffic to plasma membranes. Subsequently, we investigate the effect of N-glycosylation on the function of HCN2 channels. We developed a fluorescence-based flux assay, which makes it possible to establish a negative potential inside liposomes to open HCN2 channels. Using this flux assay, we demonstrate that glycosylation-defective N380Q HCN2 channels reconstituted into liposomes function similarly to WT HCN2 channels. This suggests that N-glycosylation is not required for HCN2 channels to function.

  6. Sensory deprivation regulates the development of the hyperpolarization-activated current in auditory brainstem neurons.

    PubMed

    Hassfurth, Benjamin; Magnusson, Anna K; Grothe, Benedikt; Koch, Ursula

    2009-10-01

    Hyperpolarization-activated and cyclic nucleotide-gated (HCN) channels are highly expressed in the superior olivary complex, the primary locus for binaural information processing. This hyperpolarization-activated current (I(h)) regulates the excitability of neurons and enhances the temporally precise analysis of the binaural acoustic cues. By using the whole-cell patch-clamp technique, we examined the properties of I(h) current in neurons of the lateral superior olive (LSO) and the medial nucleus of the trapezoid body (MNTB) before and after hearing onset. Moreover, we tested the hypothesis that I(h) currents are actively regulated by sensory input activity by performing bilateral and unilateral cochlear ablations before hearing onset, resulting in a chronic auditory deprivation. The results show that after hearing onset, I(h) currents are rapidly upregulated in LSO neurons, but change only marginally in neurons of the MNTB. We also found a striking difference in maximal current density, voltage dependence and activation time constant between the LSO and the MNTB in mature-like animals. Following bilateral cochlear ablations before hearing onset, the I(h) currents were scaled up in the LSO and scaled down in the MNTB. Consequently, in the LSO this resulted in a depolarized resting membrane potential and a lower input resistance of these neurons. This type of activity-dependent homeostatic change could thus result in an augmented response to the remaining inputs.

  7. High-throughput hyperpolarized 13C metabolic investigations using a multi-channel acquisition system

    NASA Astrophysics Data System (ADS)

    Lee, Jaehyuk; Ramirez, Marc S.; Walker, Christopher M.; Chen, Yunyun; Yi, Stacey; Sandulache, Vlad C.; Lai, Stephen Y.; Bankson, James A.

    2015-11-01

    Magnetic resonance imaging and spectroscopy of hyperpolarized (HP) compounds such as [1-13C]-pyruvate have shown tremendous potential for offering new insight into disease and response to therapy. New applications of this technology in clinical research and care will require extensive validation in cells and animal models, a process that may be limited by the high cost and modest throughput associated with dynamic nuclear polarization. Relatively wide spectral separation between [1-13C]-pyruvate and its chemical endpoints in vivo are conducive to simultaneous multi-sample measurements, even in the presence of a suboptimal global shim. Multi-channel acquisitions could conserve costs and accelerate experiments by allowing acquisition from multiple independent samples following a single dissolution. Unfortunately, many existing preclinical MRI systems are equipped with only a single channel for broadband acquisitions. In this work, we examine the feasibility of this concept using a broadband multi-channel digital receiver extension and detector arrays that allow concurrent measurement of dynamic spectroscopic data from ex vivo enzyme phantoms, in vitro anaplastic thyroid carcinoma cells, and in vivo in tumor-bearing mice. Throughput and the cost of consumables were improved by up to a factor of four. These preliminary results demonstrate the potential for efficient multi-sample studies employing hyperpolarized agents.

  8. Hyperpolarization-activated, cyclic nucleotide-gated cation channels in Aplysia: Contribution to classical conditioning

    PubMed Central

    Yang, Qizong; Kuzyk, Pavlo; Antonov, Igor; Bostwick, Caleb J.; Kohn, Andrea B.; Moroz, Leonid L.; Hawkins, Robert D.

    2015-01-01

    Hyperpolarization-activated, cyclic nucleotide-gated cation (HCN) channels are critical regulators of neuronal excitability, but less is known about their possible roles in synaptic plasticity and memory circuits. Here, we characterized the HCN gene organization, channel properties, distribution, and involvement in associative and nonassociative forms of learning in Aplysia californica. Aplysia has only one HCN gene, which codes for a channel that has many similarities to the mammalian HCN channel. The cloned acHCN gene was expressed in Xenopus oocytes, which displayed a hyperpolarization-induced inward current that was enhanced by cGMP as well as cAMP. Similarly to its homologs in other animals, acHCN is permeable to K+ and Na+ ions, and is selectively blocked by Cs+ and ZD7288. We found that acHCN is predominantly expressed in inter- and motor neurons, including LFS siphon motor neurons, and therefore tested whether HCN channels are involved in simple forms of learning of the siphon-withdrawal reflex in a semiintact preparation. ZD7288 (100 μM) significantly reduced an associative form of learning (classical conditioning) but had no effect on two nonassociative forms of learning (intermediate-term sensitization and unpaired training) or baseline responses. The HCN current is enhanced by nitric oxide (NO), which may explain the postsynaptic role of NO during conditioning. HCN current in turn enhances the NMDA-like current in the motor neurons, suggesting that HCN channels contribute to conditioning through this pathway. PMID:26668355

  9. Hyperpolarization-independent maturation and refinement of GABA/glycinergic connections in the auditory brain stem

    PubMed Central

    Lee, Hanmi; Bach, Eva; Noh, Jihyun; Delpire, Eric

    2015-01-01

    During development GABA and glycine synapses are initially excitatory before they gradually become inhibitory. This transition is due to a developmental increase in the activity of neuronal potassium-chloride cotransporter 2 (KCC2), which shifts the chloride equilibrium potential (ECl) to values more negative than the resting membrane potential. While the role of early GABA and glycine depolarizations in neuronal development has become increasingly clear, the role of the transition to hyperpolarization in synapse maturation and circuit refinement has remained an open question. Here we investigated this question by examining the maturation and developmental refinement of GABA/glycinergic and glutamatergic synapses in the lateral superior olive (LSO), a binaural auditory brain stem nucleus, in KCC2-knockdown mice, in which GABA and glycine remain depolarizing. We found that many key events in the development of synaptic inputs to the LSO, such as changes in neurotransmitter phenotype, strengthening and elimination of GABA/glycinergic connection, and maturation of glutamatergic synapses, occur undisturbed in KCC2-knockdown mice compared with wild-type mice. These results indicate that maturation of inhibitory and excitatory synapses in the LSO is independent of the GABA and glycine depolarization-to-hyperpolarization transition. PMID:26655825

  10. A Molecular Imaging Approach to Mercury Sensing Based on Hyperpolarized (129)Xe Molecular Clamp Probe.

    PubMed

    Guo, Qianni; Zeng, Qingbin; Jiang, Weiping; Zhang, Xiaoxiao; Luo, Qing; Zhang, Xu; Bouchard, Louis-S; Liu, Maili; Zhou, Xin

    2016-03-14

    Mercury pollution, in the form of mercury ions (Hg(2+)), is a major health and environmental hazard. Commonly used sensors are invasive and limited to point measurements. Fluorescence-based sensors do not provide depth resolution needed to image spatial distributions. Herein we report a novel sensor capable of yielding spatial distributions by MRI using hyperpolarized (129)Xe. A molecular clamp probe was developed consisting of dipyrrolylquinoxaline (DPQ) derivatives and twocryptophane-A cages. The DPQ derivatives act as cation receptors whereas cryptophane-A acts as a suitable host molecule for xenon. When the DPQ moiety interacts with mercury ions, the molecular clamp closes on the ion. Due to overlap of the electron clouds of the two cryptophane-A cages, the shielding effect on the encapsulated Xe becomes important. This leads to an upfield change of the chemical shift of the encapsulated Xe. This sensor exhibits good selectivity and sensitivity toward the mercury ion. This mercury-activated hyperpolarized (129)Xe-based chemosensor is a new concept method for monitoring Hg(2+) ion distributions by MRI.

  11. Hyperpolarized noble gas magnetic resonance imaging of the animal lung: Approaches and applications

    NASA Astrophysics Data System (ADS)

    Santyr, Giles E.; Lam, Wilfred W.; Parra-Robles, Juan M.; Taves, Timothy M.; Ouriadov, Alexei V.

    2009-05-01

    Hyperpolarized noble gas (HNG) magnetic resonance (MR) imaging is a very promising noninvasive tool for the investigation of animal models of lung disease, particularly to follow longitudinal changes in lung function and anatomy without the accumulated radiation dose associated with x rays. The two most common noble gases for this purpose are H3e (helium 3) and X129e (xenon 129), the latter providing a cost-effective approach for clinical applications. Hyperpolarization is typically achieved using spin-exchange optical pumping techniques resulting in ˜10 000-fold improvement in available magnetization compared to conventional Boltzmann polarizations. This substantial increase in polarization allows high spatial resolution (<1 mm) single-slice images of the lung to be obtained with excellent temporal resolution (<1 s). Complete three-dimensional images of the lungs with 1 mm slice thickness can be obtained within reasonable breath-hold intervals (<20 s). This article provides an overview of the current methods used in HNG MR imaging with an emphasis on ventilation studies in animals. Special MR hardware and software considerations are described in order to use the strong but nonrecoverable magnetization as efficiently as possible and avoid depolarization primarily by molecular oxygen. Several applications of HNG MR imaging are presented, including measurement of gross lung anatomy (e.g., airway diameters), microscopic anatomy (e.g., apparent diffusion coefficient), and a variety of functional parameters including dynamic ventilation, alveolar oxygen partial pressure, and xenon diffusing capacity.

  12. Dynamic UltraFast 2D EXchange SpectroscopY (UF-EXSY) of hyperpolarized substrates

    PubMed Central

    Swisher, Christine Leon; Koelsch, Bertram; Sukumar, Subramianam; Sriram, Renuka; Santos, Romelyn Delos; Wang, Zhen Jane; Kurhanewicz, John; Vigneron, Daniel; Larson, Peder

    2015-01-01

    In this work, we present a new ultrafast method for acquiring dynamic 2D EXchange SpectroscopY (EXSY) within a single acquisition. This technique reconstructs two-dimensional EXSY spectra from one-dimensional spectra based on the phase accrual during echo times. The Ultrafast-EXSY acquisition overcomes long acquisition times typically needed to acquire 2D NMR data by utilizing sparsity and phase dependence to dramatically undersample in the indirect time dimension. This allows for the acquisition of the 2D spectrum within a single shot. We have validated this method in simulations and hyperpolarized enzyme assay experiments separating the dehydration of pyruvate and lactate-to-pyruvate conversion. In a renal cell carcinoma cell (RCC) line, bidirectional exchange was observed. This new technique revealed decreased conversion of lactate-to-pyruvate with high expression of monocarboxylate transporter 4 (MCT4), known to correlate with aggressive cancer phenotypes. We also showed feasibility of this technique in vivo in a RCC model where bidirectional exchange was observed for pyruvate–lactate, pyruvate–alanine, and pyruvate–hydrate and were resolved in time. Broadly, the technique is well suited to investigate the dynamics of multiple exchange pathways and applicable to hyperpolarized substrates where chemical exchange has shown great promise across a range of disciplines. PMID:26117655

  13. Optimizing water hyperpolarization and dissolution for sensitivity-enhanced 2D biomolecular NMR

    NASA Astrophysics Data System (ADS)

    Olsen, Greg; Markhasin, Evgeny; Szekely, Or; Bretschneider, Christian; Frydman, Lucio

    2016-03-01

    A recent study explored the use of hyperpolarized water, to enhance the sensitivity of nuclei in biomolecules thanks to rapid proton exchanges with labile amide backbone and sidechain groups. Further optimizations of this approach have now allowed us to achieve proton polarizations approaching 25% in the water transferred into the NMR spectrometer, effective water T1 times approaching 40 s, and a reduction in the dilution demanded for the cryogenic dissolution process. Further hardware developments have allowed us to perform these experiments, repeatedly and reliably, in 5 mm NMR tubes. All these ingredients - particularly the ⩾3000× 1H polarization enhancements over 11.7 T thermal counterparts, long T1 times and a compatibility with high-resolution biomolecular NMR setups - augur well for hyperpolarized 2D NMR studies of peptides, unfolded proteins and intrinsically disordered systems undergoing fast exchanges of their protons with the solvent. This hypothesis is here explored by detailing the provisions that lead to these significant improvements over previous reports, and demonstrating 1D coherence transfer experiments and 2D biomolecular HMQC acquisitions delivering NMR spectral enhancements of 100-500× over their optimized, thermally-polarized, counterparts.

  14. Pathological impact of hyperpolarization-activated chloride current peculiar to rat pulmonary vein cardiomyocytes.

    PubMed

    Okamoto, Yosuke; Kawamura, Koichi; Nakamura, Yuta; Ono, Kyoichi

    2014-01-01

    Pulmonary veins (PVs) are believed to be a crucial origin of atrial fibrillation. We recently reported that rat PV cardiomyocytes exhibit arrhythmogenic automaticity in response to norepinephrine. Herein, we further characterized the electrophysiological properties underlying the potential arrhythmogenicity of PV cardiomyocytes. Patch clamping studies revealed a time dependent hyperpolarization-activated inward current in rat PV cardiomyocytes, but not in left atrial (LA) myocytes. The current was Cs(+) resistant, and was not affected by removal of external Na(+) or K(+). The current was inhibited with Cd(2+), and the reversal potential was sensitive to changes in [Cl(-)] on either side of the membrane in a manner consistent with a Cl(-) selective channel. Cl(-) channel blockers attenuated the current, and slowed or completely inhibited the norepinephrine-induced automaticity. The biophysical properties of the hyperpolarization-activated Cl(-) current in rat PVs were different from those of ClC-2 currents previously reported: (i) the voltage-dependent activation of the Cl(-) current in rat PVs was shifted to negative potentials as [Cl(-)]i increased, (ii) the Cl(-) current was enhanced by extracellular acidification, and (iii) extracellular hyper-osmotic stress increased the current, whereas hypo-osmotic cell swelling suppressed the current. qPCR analysis revealed negligible ClC-2 mRNA expression in the rat PV. These findings suggest that rat PV cardiomyocytes possess a peculiar voltage-dependent Cl(-) channel, and that the channel may play a functional role in norepinephrine-induced automaticity.

  15. Quantification of rate constants for successive enzymatic reactions with DNP hyperpolarized MR.

    PubMed

    Allouche-Arnon, Hyla; Hovav, Yonatan; Friesen-Waldner, Lanette; Sosna, Jacob; Moshe Gomori, J; Vega, Shimon; Katz-Brull, Rachel

    2014-06-01

    A kinetic model is provided to obtain reaction rate constants in successive enzymatic reactions that are monitored using NMR spectroscopy and hyperpolarized substrates. The model was applied for simulation and analysis of the successive oxidation of choline to betaine aldehyde, and further to betaine, by the enzyme choline oxidase. This enzymatic reaction was investigated under two different sets of conditions: two different choline molecular probes were used, [1,1,2,2-D4 , 1-(13) C]choline chloride and [1,1,2,2-D4 , 2-(13) C]choline chloride, in different MR systems (clinical scanner and high-resolution spectrometer), as well as in different reactors and reaction volumes (4.8 and 0.7 mL). The kinetic analysis according to the model yielded similar results in both set-ups, supporting the robustness of the model. This was achieved despite the complex and negating influences of reaction kinetics and polarization decay, and in the presence of uncontrolled mixing characteristics, which may introduce uncertainties in both effective timing and effective pulses. The ability to quantify rate constants using hyperpolarized MR in the first seconds of consecutive enzyme activity is important for further development of the utilization of dynamic nuclear polarization-MR for biological determinations.

  16. Dynamic UltraFast 2D EXchange SpectroscopY (UF-EXSY) of hyperpolarized substrates

    NASA Astrophysics Data System (ADS)

    Leon Swisher, Christine; Koelsch, Bertram; Sukumar, Subramianam; Sriram, Renuka; Santos, Romelyn Delos; Wang, Zhen Jane; Kurhanewicz, John; Vigneron, Daniel; Larson, Peder

    2015-08-01

    In this work, we present a new ultrafast method for acquiring dynamic 2D EXchange SpectroscopY (EXSY) within a single acquisition. This technique reconstructs two-dimensional EXSY spectra from one-dimensional spectra based on the phase accrual during echo times. The Ultrafast-EXSY acquisition overcomes long acquisition times typically needed to acquire 2D NMR data by utilizing sparsity and phase dependence to dramatically undersample in the indirect time dimension. This allows for the acquisition of the 2D spectrum within a single shot. We have validated this method in simulations and hyperpolarized enzyme assay experiments separating the dehydration of pyruvate and lactate-to-pyruvate conversion. In a renal cell carcinoma cell (RCC) line, bidirectional exchange was observed. This new technique revealed decreased conversion of lactate-to-pyruvate with high expression of monocarboxylate transporter 4 (MCT4), known to correlate with aggressive cancer phenotypes. We also showed feasibility of this technique in vivo in a RCC model where bidirectional exchange was observed for pyruvate-lactate, pyruvate-alanine, and pyruvate-hydrate and were resolved in time. Broadly, the technique is well suited to investigate the dynamics of multiple exchange pathways and applicable to hyperpolarized substrates where chemical exchange has shown great promise across a range of disciplines.

  17. Noradrenergic and serotonergic modulation of a hyperpolarization-activated cation current in thalamic relay neurones.

    PubMed Central

    McCormick, D A; Pape, H C

    1990-01-01

    1. Modulation of the hyperpolarization-activated cation current, Ih, by noradrenaline (NA) and serotonin (5-HT) was examined in guinea-pig and cat medial and lateral geniculate relay neurones using the in vitro slice technique. 2. In the absence of pharmacological antagonists, local application of NA resulted in a slow depolarization and decrease in apparent input conductance, a response which was blocked by local or bath application of the alpha 1-adrenoceptor antagonist prazosin. Application of NA after pharmacological block of alpha 1- and alpha 2-adrenoceptors, or application of 5-HT in all conditions, induced a 1-3 mV slow depolarization which was associated with a pronounced increase in apparent input conductance. This response to NA and 5-HT persisted during blocked synaptic transmission and was present in both the guinea-pig and cat medial and lateral geniculate nuclei. 3. The increase in membrane conductance elicited by NA was mimicked by the beta-specific agonist isoprenaline and blocked by the beta-antagonists propranolol and atenolol, indicating that it is mediated by beta-adrenoceptors. The response to 5-HT was blocked by the 5-HT1 and 5-HT2 antagonist methysergide, but not by the 5-HT2 antagonist ritanserin. Applications of either the 5-HT1A agonist ipsapirone or the partial agonist 8-hydroxy-dipropylaminotetralin (8-OHDPAT) were without effect. 4. Current versus voltage relationships obtained under voltage clamp revealed NA and 5-HT to cause a voltage-dependent inward shift at membrane potentials negative to approximately -60 mV. This response appeared to be shared by NA and 5-HT since maximal application of 5-HT greatly reduced or abolished the response to NA. 5. Application of NA and/or 5-HT during hyperpolarizing voltage steps in voltage clamp resulted in a marked increase in amplitude of the hyperpolarization-activated cation current, Ih. In addition, the rate of activation of Ih was strongly increased during activation of beta-adrenoceptors. 6

  18. Noradrenergic and serotonergic modulation of a hyperpolarization-activated cation current in thalamic relay neurones.

    PubMed

    McCormick, D A; Pape, H C

    1990-12-01

    1. Modulation of the hyperpolarization-activated cation current, Ih, by noradrenaline (NA) and serotonin (5-HT) was examined in guinea-pig and cat medial and lateral geniculate relay neurones using the in vitro slice technique. 2. In the absence of pharmacological antagonists, local application of NA resulted in a slow depolarization and decrease in apparent input conductance, a response which was blocked by local or bath application of the alpha 1-adrenoceptor antagonist prazosin. Application of NA after pharmacological block of alpha 1- and alpha 2-adrenoceptors, or application of 5-HT in all conditions, induced a 1-3 mV slow depolarization which was associated with a pronounced increase in apparent input conductance. This response to NA and 5-HT persisted during blocked synaptic transmission and was present in both the guinea-pig and cat medial and lateral geniculate nuclei. 3. The increase in membrane conductance elicited by NA was mimicked by the beta-specific agonist isoprenaline and blocked by the beta-antagonists propranolol and atenolol, indicating that it is mediated by beta-adrenoceptors. The response to 5-HT was blocked by the 5-HT1 and 5-HT2 antagonist methysergide, but not by the 5-HT2 antagonist ritanserin. Applications of either the 5-HT1A agonist ipsapirone or the partial agonist 8-hydroxy-dipropylaminotetralin (8-OHDPAT) were without effect. 4. Current versus voltage relationships obtained under voltage clamp revealed NA and 5-HT to cause a voltage-dependent inward shift at membrane potentials negative to approximately -60 mV. This response appeared to be shared by NA and 5-HT since maximal application of 5-HT greatly reduced or abolished the response to NA. 5. Application of NA and/or 5-HT during hyperpolarizing voltage steps in voltage clamp resulted in a marked increase in amplitude of the hyperpolarization-activated cation current, Ih. In addition, the rate of activation of Ih was strongly increased during activation of beta-adrenoceptors. 6

  19. Performance of 4600-pound-thrust centrifugal-flow-type turbojet engine with water-alcohol injection at inlet

    NASA Technical Reports Server (NTRS)

    Glasser, Philip W

    1950-01-01

    An experimental investigation of the effects of injecting a water-alcohol mixture of 2:1 at the compressor inlet of a centrifugal-flow type turbojet engine was conducted in an altitude test chamber at static sea-level conditions and at an altitude of 20,000 feet with a flight Mach number of 0.78 with an engine operating at rated speed. The net thrust was augmented by 0.16 for both flight conditions with a ratio of injected liquid to air flow of 0.05. Further increases in the liquid-air ratio did not give comparable increases in thrust.

  20. Imaging oxygen metabolism with hyperpolarized magnetic resonance: a novel approach for the examination of cardiac and renal function

    PubMed Central

    Schroeder, Marie

    2016-01-01

    Every tissue in the body critically depends on meeting its energetic demands with sufficient oxygen supply. Oxygen supply/demand imbalances underlie the diseases that inflict the greatest socio-economic burden globally. The purpose of this review is to examine how hyperpolarized contrast media, used in combination with MR data acquisition methods, may advance our ability to assess oxygen metabolism non-invasively and thus improve management of clinical disease. We first introduce the concept of hyperpolarization and how hyperpolarized contrast media have been practically implemented to achieve translational and clinical research. We will then analyse how incorporating hyperpolarized contrast media could enable realization of unmet technical needs in clinical practice. We will focus on imaging cardiac and renal oxygen metabolism, as both organs have unique physiological demands to satisfy their requirements for tissue oxygenation, their dysfunction plays a fundamental role in society’s most prevalent diseases, and each organ presents unique imaging challenges. It is our aim that this review attracts a multi-disciplinary audience and sparks collaborations that utilize an exciting, emergent technology to advance our ability to treat patients adversely affected by an oxygen supply/demand mismatch. PMID:27899435

  1. A Bloch-McConnell simulator with pharmacokinetic modeling to explore accuracy and reproducibility in the measurement of hyperpolarized pyruvate

    NASA Astrophysics Data System (ADS)

    Walker, Christopher M.; Bankson, James A.

    2015-03-01

    Magnetic resonance imaging (MRI) of hyperpolarized (HP) agents has the potential to probe in-vivo metabolism with sensitivity and specificity that was not previously possible. Biological conversion of HP agents specifically for cancer has been shown to correlate to presence of disease, stage and response to therapy. For such metabolic biomarkers derived from MRI of hyperpolarized agents to be clinically impactful, they need to be validated and well characterized. However, imaging of HP substrates is distinct from conventional MRI, due to the non-renewable nature of transient HP magnetization. Moreover, due to current practical limitations in generation and evolution of hyperpolarized agents, it is not feasible to fully experimentally characterize measurement and processing strategies. In this work we use a custom Bloch-McConnell simulator with pharmacokinetic modeling to characterize the performance of specific magnetic resonance spectroscopy sequences over a range of biological conditions. We performed numerical simulations to evaluate the effect of sequence parameters over a range of chemical conversion rates. Each simulation was analyzed repeatedly with the addition of noise in order to determine the accuracy and reproducibility of measurements. Results indicate that under both closed and perfused conditions, acquisition parameters can affect measurements in a tissue dependent manner, suggesting that great care needs to be taken when designing studies involving hyperpolarized agents. More modeling studies will be needed to determine what effect sequence parameters have on more advanced acquisitions and processing methods.

  2. Toward Biocompatible Nuclear Hyperpolarization Using Signal Amplification by Reversible Exchange: Quantitative in Situ Spectroscopy and High-Field Imaging

    PubMed Central

    2014-01-01

    Signal amplification by reversible exchange (SABRE) of a substrate and parahydrogen at a catalytic center promises to overcome the inherent insensitivity of magnetic resonance. In order to apply the new approach to biomedical applications, there is a need to develop experimental equipment, in situ quantification methods, and a biocompatible solvent. We present results detailing a low-field SABRE polarizer which provides well-controlled experimental conditions, defined spins manipulations, and which allows in situ detection of thermally polarized and hyperpolarized samples. We introduce a method for absolute quantification of hyperpolarization yield in situ by means of a thermally polarized reference. A maximum signal-to-noise ratio of ∼103 for 148 μmol of substance, a signal enhancement of 106 with respect to polarization transfer field of SABRE, or an absolute 1H-polarization level of ≈10–2 is achieved. In an important step toward biomedical application, we demonstrate 1H in situ NMR as well as 1H and 13C high-field MRI using hyperpolarized pyridine (d3) and 13C nicotinamide in pure and 11% ethanol in aqueous solution. Further increase of hyperpolarization yield, implications of in situ detection, and in vivo application are discussed. PMID:24397559

  3. In vivo and in vitro liver cancer metabolism observed with hyperpolarized [5-13C]glutamine

    NASA Astrophysics Data System (ADS)

    Cabella, C.; Karlsson, M.; Canapè, C.; Catanzaro, G.; Colombo Serra, S.; Miragoli, L.; Poggi, L.; Uggeri, F.; Venturi, L.; Jensen, P. R.; Lerche, M. H.; Tedoldi, F.

    2013-07-01

    Glutamine metabolism is, with its many links to oncogene expression, considered a crucial step in cancer metabolism and it is thereby a key target for alteration in cancer development. In particular, strong correlations have been reported between oncogene expression and expression and activity of the enzyme glutaminase. This mitochondrial enzyme, which is responsible for the deamidation of glutamine to form glutamate, is overexpressed in many tumour tissues. In animal models, glutaminase expression is correlated with tumour growth rate and it is readily possible to limit tumour growth by suppression of glutaminase activity. In principle, hyperpolarized 13C MR spectroscopy can provide insight to glutamine metabolism and should hence be a valuable tool to study changes in glutaminase activity as tumours progress. However, no such successful in vivo studies have been reported, even though several good biological models have been tested. This may, at least partly, be due to problems in preparing glutamine for hyperpolarization. This paper reports a new and improved preparation of hyperpolarized [5-13C]glutamine, which provides a highly sensitive 13C MR marker. With this preparation of hyperpolarized [5-13C]glutamine, glutaminase activity in vivo in a rat liver tumour was investigated. Moreover, this marker was also used to measure response to drug treatment in vitro in cancer cells. These examples of [5-13C]glutamine used in tumour models warrant the new preparation to allow metabolic studies with this conditionally essential amino acid.

  4. Use of a Multi-compartment Dynamic Single Enzyme Phantom for Studies of Hyperpolarized Magnetic Resonance Agents.

    PubMed

    Walker, Christopher M; Merritt, Matthew; Wang, Jian-Xiong; Bankson, James A

    2016-04-15

    Imaging of hyperpolarized substrates by magnetic resonance shows great clinical promise for assessment of critical biochemical processes in real time. Due to fundamental constraints imposed by the hyperpolarized state, exotic imaging and reconstruction techniques are commonly used. A practical system for characterization of dynamic, multi-spectral imaging methods is critically needed. Such a system must reproducibly recapitulate the relevant chemical dynamics of normal and pathological tissues. The most widely utilized substrate to date is hyperpolarized [1-(13)C]-pyruvate for assessment of cancer metabolism. We describe an enzyme-based phantom system that mediates the conversion of pyruvate to lactate. The reaction is initiated by injection of the hyperpolarized agent into multiple chambers within the phantom, each of which contains varying concentrations of reagents that control the reaction rate. Multiple compartments are necessary to ensure that imaging sequences faithfully capture the spatial and metabolic heterogeneity of tissue. This system will aid the development and validation of advanced imaging strategies by providing chemical dynamics that are not available from conventional phantoms, as well as control and reproducibility that is not possible in vivo.

  5. Hyperpolarized 13C NMR lifetimes in the liquid-state: relating structures and T1 relaxation times

    NASA Astrophysics Data System (ADS)

    Parish, Christopher; Niedbalski, Peter; Hashami, Zohreh; Fidelino, Leila; Kovacs, Zoltan; Lumata, Lloyd

    Among the various attempts to solve the insensitivity problem in nuclear magnetic resonance (NMR), the physics-based technique dissolution dynamic nuclear polarization (DNP) is probably the most successful method of hyperpolarization or amplifying NMR signals. Using this technique, liquid-state NMR signal enhancements of several thousand-fold are expected for low-gamma nuclei such as carbon-13. The lifetimes of these hyperpolarized 13C NMR signals are directly related to their 13C spin-lattice relaxation times T1. Depending upon the 13C isotopic location, the lifetimes of hyperpolarized 13C compounds can range from a few seconds to minutes. In this study, we have investigated the hyperpolarized 13C NMR lifetimes of several 13C compounds with various chemical structures from glucose, acetate, citric acid, naphthalene to tetramethylallene and their deuterated analogs at 9.4 T and 25 deg C. Our results show that the 13C T1s of these compounds can range from a few seconds to more than 60 s at this field. Correlations between the chemical structures and T1 relaxation times will be discussed and corresponding implications of these results on 13C DNP experiments will be revealed. US Dept of Defense Award No. W81XWH-14-1-0048 and Robert A. Welch Foundation Grant No. AT-1877.

  6. Excitatory postsynaptic potentials trigger a plateau potential in rat subthalamic neurons at hyperpolarized states.

    PubMed

    Otsuka, T; Murakami, F; Song, W J

    2001-10-01

    The subthalamic nucleus (STN) directly innervates the output structures of the basal ganglia, playing a key role in basal ganglia function. It is therefore important to understand the regulatory mechanisms for the activity of STN neurons. In the present study, we aimed to investigate how the intrinsic membrane properties of STN neurons interact with their synaptic inputs, focusing on their generation and the properties of the long-lasting, plateau potential. Whole cell recordings were obtained from STN neurons in slices prepared from postnatal day 14 (P14) to P20 rats. We found that activation of glutamate receptor-mediated excitatory synaptic potentials (EPSPs) evoked a plateau potential in a subpopulation of STN neurons (n = 13/22), in a voltage-dependent manner. Plateau potentials could be induced only when the cell was hyperpolarized to more negative than about -75 mV. Plateau potentials, evoked with a depolarizing current pulse, again only from a hyperpolarized state, were observed in about half of STN neurons tested (n = 162/327). Only in neurons in which a plateau potential could be evoked by current injection did EPSPs evoke plateau potentials. L-type Ca(2+) channels, Ca(2+)-dependent K(+) channels, and TEA-sensitive K(+) channels were found to be involved in the generation of the potential. The stability of the plateau potential, tested by the injection of a negative pulse current during the plateau phase, was found to be robust at the early phase of the potential, but decreased toward the end. As a result the early part of the plateau potential was resistant to membrane potential perturbations and would be able to support a train of action potentials. We conclude that excitatory postsynaptic potentials, evoked in a subpopulation of STN neurons at a hyperpolarized state, activate L-type Ca(2+) and other channels, leading to the generation of a plateau potential. Thus about half of STN neurons can transform short-lasting synaptic excitation into a long

  7. Remodeling of Hyperpolarization-Activated Current, Ih, in Ah-Type Visceral Ganglion Neurons Following Ovariectomy in Adult Rats

    PubMed Central

    Xu, Wen-Xiao; Yan, Zhen-Yu; Liu, Yang; Zhou, Jia-Ying; Zhang, Hao-Cheng; Wang, Li-Juan; Pan, Xiao-Dong; Fu, Yili

    2013-01-01

    Hyperpolarization-activated currents (Ih) mediated by hyperpolarization-activated cyclic nucleotide-gated (HCN) channels modulate excitability of myelinated A− and Ah-type visceral ganglion neurons (VGN). Whether alterations in Ih underlie the previously reported reduction of excitability of myelinated Ah-type VGNs following ovariectomy (OVX) has remained unclear. Here we used the intact nodose ganglion preparation in conjunction with electrophysiological approaches to examine the role of Ih remodeling in altering Ah-type neuron excitability following ovariectomy in adult rats. Ah-type neurons were identified based on their afferent conduction velocity. Ah-type neurons in nodose ganglia from non-OVX rats exhibited a voltage ‘sag’ as well as ‘rebound’ action potentials immediately following hyperpolarizing current injections, which both were suppressed by the Ih blocker ZD7288. Repetitive spike activity induced afterhyperpolarizations lasting several hundreds of milliseconds (termed post-excitatory membrane hyperpolarizations, PEMHs), which were significantly reduced by ZD7288, suggesting that they resulted from transient deactivation of Ih during the preceding spike trains. Ovariectomy reduced whole-cell Ih density, caused a hyperpolarizing shift of the voltage-dependence of Ih activation, and slowed Ih activation. OVX-induced Ih remodeling was accompanied by a flattening of the stimulus frequency/response curve and loss of PEMHs. Also, HCN1 mRNA levels were reduced by ∼30% in nodose ganglia from OVX rats compared with their non-OVX counterparts. Acute exposure of nodose ganglia to 17beta-estradiol partly restored Ih density and accelerated Ih activation in Ah-type cells. In conclusion, Ih plays a significant role in modulating the excitability of myelinated Ah-type VGNs in adult female rats. PMID:23951107

  8. Efficient room-temperature nuclear spin hyperpolarization of a defect atom in a semiconductor.

    PubMed

    Puttisong, Y; Wang, X J; Buyanova, I A; Geelhaar, L; Riechert, H; Ptak, A J; Tu, C W; Chen, W M

    2013-01-01

    Nuclear spin hyperpolarization is essential to future solid-state quantum computation using nuclear spin qubits and in highly sensitive magnetic resonance imaging. Though efficient dynamic nuclear polarization in semiconductors has been demonstrated at low temperatures for decades, its realization at room temperature is largely lacking. Here we demonstrate that a combined effect of efficient spin-dependent recombination and hyperfine coupling can facilitate strong dynamic nuclear polarization of a defect atom in a semiconductor at room temperature. We provide direct evidence that a sizeable nuclear field (~150 Gauss) and nuclear spin polarization (~15%) sensed by conduction electrons in GaNAs originates from dynamic nuclear polarization of a Ga interstitial defect. We further show that the dynamic nuclear polarization process is remarkably fast and is completed in <5 μs at room temperature. The proposed new concept could pave a way to overcome a major obstacle in achieving strong dynamic nuclear polarization at room temperature, desirable for practical device applications.

  9. In vivo detection of brain Krebs cycle intermediate by hyperpolarized magnetic resonance.

    PubMed

    Mishkovsky, Mor; Comment, Arnaud; Gruetter, Rolf

    2012-12-01

    The Krebs (or tricarboxylic acid (TCA)) cycle has a central role in the regulation of brain energy regulation and metabolism, yet brain TCA cycle intermediates have never been directly detected in vivo. This study reports the first direct in vivo observation of a TCA cycle intermediate in intact brain, namely, 2-oxoglutarate, a key biomolecule connecting metabolism to neuronal activity. Our observation reveals important information about in vivo biochemical processes hitherto considered undetectable. In particular, it provides direct evidence that transport across the inner mitochondria membrane is rate limiting in the brain. The hyperpolarized magnetic resonance protocol designed for this study opens the way to direct and real-time studies of TCA cycle kinetics.

  10. Real-time Detection of Polymerization Reactions with Hyperpolarized Xenon at Low Magnetic Fields

    NASA Astrophysics Data System (ADS)

    Glöggler, Stefan; Blümich, Bernhard; Appelt, Stephan

    2011-03-01

    For process control it is desirable to develop simple devices for studying polymerization reactions in real-time and in-situ. We are demonstrating an approach using NMR at fields as low as 35 G and hyperpolarized xenon, which allows us to observe polymerization reactions in real-time. The investigated reaction is a free radical polymerization with the initiator azobisisobutyronitrile (AIBN) and the monomer methyl methacrylate (MMA). AIBN and MMA are mixed together in a sample tube under noble gas atmosphere, and the reaction is started by irradiation with UV light (360 nm). As the reaction goes on, xenon NMR spectra are acquired. They show increasing line broadening and a variation of the chemical shift depending on the state of polymerization. This observation gives rise to the idea that a single-sided high resolution NMR sensor can be developed with which at least light induced polymerization reactions can be studied in-situ and in real-time.

  11. Hyperpolarization of Frozen Hydrocarbon Gases by Dynamic Nuclear Polarization at 1.2 K.

    PubMed

    Vuichoud, Basile; Canet, Estel; Milani, Jonas; Bornet, Aurélien; Baudouin, David; Veyre, Laurent; Gajan, David; Emsley, Lyndon; Lesage, Anne; Copéret, Christophe; Thieuleux, Chloé; Bodenhausen, Geoffrey; Koptyug, Igor; Jannin, Sami

    2016-08-18

    We report a simple and general method for the hyperpolarization of condensed gases by dynamic nuclear polarization (DNP). The gases are adsorbed in the pores of structured mesoporous silica matrices known as HYPSOs (HYper Polarizing SOlids) that have paramagnetic polarizing agents covalently bound to the surface of the mesopores. DNP is performed at low temperatures and moderate magnetic fields (T = 1.2 K and B0 = 6.7 T). Frequency-modulated microwave irradiation is applied close to the electron spin resonance frequency (f = 188.3 GHz), and the electron spin polarization of the polarizing agents of HYPSO is transferred to the nuclear spins of the frozen gas. A proton polarization as high as P((1)H) = 70% can be obtained, which can be subsequently transferred to (13)C in natural abundance by cross-polarization, yielding up to P((13)C) = 27% for ethylene.

  12. MRI of the lung gas-space at very low-field using hyperpolarized noble gases

    NASA Technical Reports Server (NTRS)

    Venkatesh, Arvind K.; Zhang, Adelaide X.; Mansour, Joey; Kubatina, Lyubov; Oh, Chang Hyun; Blasche, Gregory; Selim Unlu, M.; Balamore, Dilip; Jolesz, Ferenc A.; Goldberg, Bennett B.; Albert, Mitchell S.

    2003-01-01

    In hyperpolarized (HP) noble-gas magnetic resonance imaging, large nuclear spin polarizations, about 100,000 times that ordinarily obtainable at thermal equilibrium, are created in 3He and 129Xe. The enhanced signal that results can be employed in high-resolution MRI studies of void spaces such as in the lungs. In HP gas MRI the signal-to-noise ratio (SNR) depends only weakly on the static magnetic field (B(0)), making very low-field (VLF) MRI possible; indeed, it is possible to contemplate portable MRI using light-weight solenoids or permanent magnets. This article reports the first in vivo VLF MR images of the lungs in humans and in rats, obtained at a field of only 15 millitesla (150 Gauss).

  13. Hyperpolarized 83Kr magnetic resonance imaging of alveolar degradation in a rat model of emphysema.

    PubMed

    Lilburn, David M L; Lesbats, Clémentine; Six, Joseph S; Dubuis, Eric; Yew-Booth, Liang; Shaw, Dominick E; Belvisi, Maria G; Birrell, Mark A; Pavlovskaya, Galina E; Meersmann, Thomas

    2015-06-06

    Hyperpolarized (83)Kr surface quadrupolar relaxation (SQUARE) generates MRI contrast that was previously shown to correlate with surface-to-volume ratios in porous model surface systems. The underlying physics of SQUARE contrast is conceptually different from any other current MRI methodology as the method uses the nuclear electric properties of the spin I = 9/2 isotope (83)Kr. To explore the usage of this non-radioactive isotope for pulmonary pathophysiology, MRI SQUARE contrast was acquired in excised rat lungs obtained from an elastase-induced model of emphysema. A significant (83)Kr T1 relaxation time increase in the SQUARE contrast was found in the elastase-treated lungs compared with the baseline data from control lungs. The SQUARE contrast suggests a reduction in pulmonary surface-to-volume ratio in the emphysema model that was validated by histology. The finding supports usage of (83)Kr SQUARE as a new biomarker for surface-to-volume ratio changes in emphysema.

  14. Quadrupolar relaxation of hyperpolarized krypton-83 as a probe for surfaces.

    PubMed

    Stupic, Karl F; Cleveland, Zackary I; Pavlovskaya, Galina E; Meersmann, Thomas

    2006-02-01

    This work reports the first systematic study of relaxation experienced by the hyperpolarized (hp) noble gas isotope (83)Kr (I=9/2) in contact with surfaces. The spin-lattice relaxation of (83)Kr is found to depend strongly on the chemical composition of the surfaces in the vicinity of the gas. This effect is caused by quadrupolar interactions during brief periods of surface adsorption that are the dominating source of longitudinal spin relaxation in the (83)Kr atoms. Simple model systems of closest packed glass beads with uniform but variable bead sizes are used for the relaxation measurements. The observed relaxation rates depend strongly on the chemical treatment of the glass surfaces and on the surface to volume ratio. Hp (83)Kr NMR relaxation measurements of porous polymers with pore sizes of 70-250 microm demonstrate the potential use of this new technique for material sciences applications.

  15. Hyperpolarized krypton-83 as a contrast agent for magnetic resonance imaging.

    PubMed

    Pavlovskaya, Galina E; Cleveland, Zackary I; Stupic, Karl F; Basaraba, Randall J; Meersmann, Thomas

    2005-12-20

    For the first time, magnetic resonance imaging (MRI) with hyperpolarized (hp) krypton-83 (83Kr) has become available. The relaxation of the nuclear spin of 83Kr atoms (I = 9/2) is driven by quadrupolar interactions during brief adsorption periods on surrounding material interfaces. Experiments in model systems reveal that the longitudinal relaxation of hp 83Kr gas strongly depends on the chemical composition of the materials. The relaxation-weighted contrast in hp 83Kr MRI allows for the distinction between hydrophobic and hydrophilic surfaces. The feasibility of hp 83Kr MRI of airways is tested in canine lung tissue by using krypton gas with natural abundance isotopic distribution. Additionally, the influence of magnetic field strength and the presence of a breathable concentration of molecular oxygen on longitudinal relaxation are investigated.

  16. Fast Volumetric Spatial-Spectral MR Imaging of Hyperpolarized 13C-Labeled Compounds using Multiple Echo 3D bSSFP

    PubMed Central

    Perman, William H.; Bhattacharya, Pratip; Leupold, Jochen; Lin, Alexander P.; Harris, Kent C.; Norton, Valerie A.; Hovener, Jan B.; Ross, Brian D.

    2010-01-01

    PURPOSE The goal of this work was to develop a fast 3D chemical shift imaging technique for the non-invasive measurement of hyperpolarized 13C-labeled substrates and metabolic products at low concentration. MATERIALS AND METHODS Multiple echo 3D balanced steady state MR imaging (ME-3DbSSFP) was performed in vitro on a syringe containing hyperpolarized [1,3,3-2H3; 1-13C]2-hydroxyethylpropionate (HEP) adjacent to a 13C-enriched acetate phantom, and in vivo on a rat before and after IV injection of hyperpolarized HEP at 1.5 T. Chemical shift images of the hyperpolarized HEP were derived from the multiple echo data by Fourier transformation along the echoes on a voxel by voxel basis for each slice of the 3D data set. RESULTS ME-3DbSSFP imaging was able to provide chemical shift images of hyperpolarized HEP in vivo, and in a rat with isotropic 7 mm spatial resolution, 93 Hz spectral resolution and 16 second temporal resolution for a period greater than 45 seconds. CONCLUSION Multiple echo 3D bSSFP imaging can provide chemical shift images of hyperpolarized 13C-labeled compounds in vivo with relatively high spatial resolution and moderate spectral resolution. The increased signal-to-noise ratio (SNR) of this 3D technique will enable the detection of hyperpolarized 13C-labeled metabolites at lower concentrations as compared to a 2D technique. PMID:20171034

  17. Components of after-hyperpolarization in magnocellular neurones of the rat supraoptic nucleus in vitro

    PubMed Central

    Greffrath, Wolfgang; Martin, Erich; Reuss, Stefan; Boehmer, Gerd

    1998-01-01

    The pharmacological sensitivity of hyperpolarizing components of spike train after-potentials was examined in sixty-one magnocellular neurones of the rat supraoptic nucleus using intracellular recording techniques in a brain slice preparation.In 26 % of all neurones a slow after-hyperpolarization (AHP) was observed in addition to a fast AHP. In 31 % of all neurones a depolarizing after-potential (DAP) was observed.The fast AHP was blocked by apamin whereas the slow AHP was blocked by charybdotoxin (ChTX). The DAP was enhanced by ChTX or a DAP was unmasked if not present during the control period.Low concentrations of TEA (0.15–1.5 mm) induced effects on the slow AHP and the DAP essentially resembling those of ChTX. The same was true for the effects of CoCl2 (1 mm).Spike train after-potentials were not affected by either iberiotoxin (IbTX), a selective high-conductance potassium (BK) channel antagonist, or margatoxin (MgTX), a Kv1.3 α-subunit antagonist.Kv1.3 α-subunit immunohistochemistry revealed that these units are not expressed in the somato-dendritic region of supraoptic neurones.The effects of ChTX, IbTX, MgTX, TEA, CoCl2 and CdCl2 on spike train after-potentials are interpreted in terms of an induction of the slow AHP by the activation of calcium-dependent potassium channels of intermediate single channel conductance (IK channels).The results suggest that at least the majority of supraoptic magnocellular neurones share the capability of generating both a slow AHP and a DAP. The slow AHP may act to control the expression of the DAP, thus regulating the excitability of magnocellular neurones. The interaction of the slow AHP and the DAP may be important for the control of phasic discharge. PMID:9806998

  18. Simultaneous in vivo assessment of cardiac and hepatic metabolism in the diabetic rat using hyperpolarized MRS

    PubMed Central

    Ball, Daniel R.; Ball, Vicky; Dodd, Michael S.; Miller, Jack J.; Heather, Lisa C.; Tyler, Damian J.

    2016-01-01

    Understanding and assessing diabetic metabolism is vital for monitoring disease progression and improving treatment of patients. In vivo assessments, using MRI and MRS, provide non‐invasive and accurate measurements, and the development of hyperpolarized 13C spectroscopy in particular has been demonstrated to provide valuable metabolic data in real time. Until now, studies have focussed on individual organs. However, diabetes is a systemic disease affecting multiple tissues in the body. Therefore, we have developed a technique to simultaneously measure metabolism in both the heart and liver during a single acquisition. A hyperpolarized 13C MRS protocol was developed to allow acquisition of metabolic data from the heart and liver during a single scan. This protocol was subsequently used to assess metabolism in the heart and liver of seven control male Wistar rats and seven diabetic rats (diabetes was induced by three weeks of high‐fat feeding and a 30 mg/kg injection of streptozotocin). Using our new acquisition, we observed decreased cardiac and hepatic pyruvate dehydrogenase flux in our diabetic rat model. These diabetic rats also had increased blood glucose levels, decreased insulin, and increased hepatic triglycerides. Decreased production of hepatic [1‐13C]alanine was observed in the diabetic group, but this change was not present in the hearts of the same diabetic animals. We have demonstrated the ability to measure cardiac and hepatic metabolism simultaneously, with sufficient sensitivity to detect metabolic alterations in both organs. Further, we have non‐invasively observed the different reactions of the heart and liver to the metabolic challenge of diabetes. PMID:27779334

  19. Hyperpolarized Functional Magnetic Resonance of Murine Skeletal Muscle Enabled by Multiple Tracer-Paradigm Synchronizations

    PubMed Central

    Leftin, Avigdor; Roussel, Tangi; Frydman, Lucio

    2014-01-01

    Measuring metabolism's time- and space-dependent responses upon stimulation lies at the core of functional magnetic resonance imaging. While focusing on water's sole resonance, further insight could arise from monitoring the temporal responses arising from the metabolites themselves, in what is known as functional magnetic resonance spectroscopy. Performing these measurements in real time, however, is severely challenged by the short functional timescales and low concentrations of natural metabolites. Dissolution dynamic nuclear polarization is an emerging technique that can potentially alleviate this, as it provides a massive sensitivity enhancement allowing one to probe low-concentration tracers and products in a single-scan. Still, conventional implementations of this hyperpolarization approach are not immediately amenable to the repeated acquisitions needed in real-time functional settings. This work proposes a strategy for functional magnetic resonance of hyperpolarized metabolites that bypasses this limitation, and enables the observation of real-time metabolic changes through the synchronization of stimuli-triggered, multiple-bolus injections of the metabolic tracer 13C1-pyruvate. This new approach is demonstrated with paradigms tailored to reveal in vivo thresholds of murine hind-limb skeletal muscle activation, involving the conversion of 13C1-pyruvate to 13C1-lactate and 13C1-alanine. These functional hind-limb studies revealed that graded skeletal muscle stimulation causes commensurate increases in glycolytic metabolism in a frequency- and amplitude-dependent fashion, that can be monitored on the seconds/minutes timescale using dissolution dynamic nuclear polarization. Spectroscopic imaging further allowed the in vivo visualization of uptake, transformation and distribution of the tracer and products, in fast-twitch glycolytic and in slow-twitch oxidative muscle fiber groups. While these studies open vistas in time and sensitivity for metabolic

  20. Endothelium-derived hyperpolarizing factor contributes to hypoxia-induced skeletal muscle vasodilation in humans.

    PubMed

    Spilk, Samson; Herr, Michael D; Sinoway, Lawrence I; Leuenberger, Urs A

    2013-12-01

    Systemic hypoxia causes skeletal muscle vasodilation, thereby preserving O2 delivery to active tissues. Nitric oxide (NO), adenosine, and prostaglandins contribute to this vasodilation, but other factors may also play a role. We tested the hypothesis that regional inhibition of endothelium-derived hyperpolarizing factor with the cytochrome P-450 2C9 antagonist fluconazole, alone or combined with the NO synthase antagonist N(G)-monomethyl-L-arginine (L-NMMA), attenuates hypoxia-induced vasodilation. We compared forearm blood flow (FBF) and skin blood flow before and during brachial artery infusion of fluconazole (0.3 mg/min; trial 1) or fluconazole + L-NMMA (50 mg over 10 min; trial 2) and during systemic hypoxia (10 min, arterial Po2 ~37 mmHg) in infused (experimental) and control forearms of 12 healthy humans. During normoxia, fluconazole and fluconazole + L-NMMA reduced (P < 0.05) forearm vascular conductance (FVC) by ~10% and ~18%, respectively. During hypoxia and fluconazole (trial 1), FVC increased by 1.76 ± 0.37 and 0.95 ± 0.35 units in control and experimental forearms, respectively (P < 0.05). During hypoxia and fluconazole + L-NMMA (trial 2), FVC increased by 2.32 ± 0.51 and 0.72 ± 0.22 units in control and experimental forearms, respectively (P < 0.05). Similarly, during hypoxia with L-NMMA alone (trial 3; n = 8) FVC increased by 1.51 ± 0.46 and 0.45 ± 0.32 units in control and experimental forearms, respectively (P < 0.05). These effects were not due to altered skin blood flow. We conclude that endothelium-derived hyperpolarizing factor contributes to basal vascular tone and to hypoxia-induced skeletal muscle vasodilation and could be particularly relevant when other vasodilator systems are impaired.

  1. Hyperpolarized functional magnetic resonance of murine skeletal muscle enabled by multiple tracer-paradigm synchronizations.

    PubMed

    Leftin, Avigdor; Roussel, Tangi; Frydman, Lucio

    2014-01-01

    Measuring metabolism's time- and space-dependent responses upon stimulation lies at the core of functional magnetic resonance imaging. While focusing on water's sole resonance, further insight could arise from monitoring the temporal responses arising from the metabolites themselves, in what is known as functional magnetic resonance spectroscopy. Performing these measurements in real time, however, is severely challenged by the short functional timescales and low concentrations of natural metabolites. Dissolution dynamic nuclear polarization is an emerging technique that can potentially alleviate this, as it provides a massive sensitivity enhancement allowing one to probe low-concentration tracers and products in a single-scan. Still, conventional implementations of this hyperpolarization approach are not immediately amenable to the repeated acquisitions needed in real-time functional settings. This work proposes a strategy for functional magnetic resonance of hyperpolarized metabolites that bypasses this limitation, and enables the observation of real-time metabolic changes through the synchronization of stimuli-triggered, multiple-bolus injections of the metabolic tracer 13C1-pyruvate. This new approach is demonstrated with paradigms tailored to reveal in vivo thresholds of murine hind-limb skeletal muscle activation, involving the conversion of 13C1-pyruvate to 13C1-lactate and 13C1-alanine. These functional hind-limb studies revealed that graded skeletal muscle stimulation causes commensurate increases in glycolytic metabolism in a frequency- and amplitude-dependent fashion, that can be monitored on the seconds/minutes timescale using dissolution dynamic nuclear polarization. Spectroscopic imaging further allowed the in vivo visualization of uptake, transformation and distribution of the tracer and products, in fast-twitch glycolytic and in slow-twitch oxidative muscle fiber groups. While these studies open vistas in time and sensitivity for metabolic

  2. Reaction monitoring using hyperpolarized NMR with scaling of heteronuclear couplings by optimal tracking.

    PubMed

    Zhang, Guannan; Schilling, Franz; Glaser, Steffen J; Hilty, Christian

    2016-11-01

    Off-resonance decoupling using the method of Scaling of Heteronuclear Couplings by Optimal Tracking (SHOT) enables determination of heteronuclear correlations of chemical shifts in single scan NMR spectra. Through modulation of J-coupling evolution by shaped radio frequency pulses, off resonance decoupling using SHOT pulses causes a user-defined dependence of the observed J-splitting, such as the splitting of (13)C peaks, on the chemical shift offset of coupled nuclei, such as (1)H. Because a decoupling experiment requires only a single scan, this method is suitable for characterizing on-going chemical reactions using hyperpolarization by dissolution dynamic nuclear polarization (D-DNP). We demonstrate the calculation of [(13)C, (1)H] chemical shift correlations of the carbanionic active sites from hyperpolarized styrene polymerized using sodium naphthalene as an initiator. While off resonance decoupling by SHOT pulses does not enhance the resolution in the same way as a 2D NMR spectrum would, the ability to obtain the correlations in single scans makes this method ideal for determination of chemical shifts in on-going reactions on the second time scale. In addition, we present a novel SHOT pulse that allows to scale J-splittings 50% larger than the respective J-coupling constant. This feature can be used to enhance the resolution of the indirectly detected chemical shift and reduce peak overlap, as demonstrated in a model reaction between p-anisaldehyde and isobutylamine. For both pulses, the accuracy is evaluated under changing signal-to-noise ratios (SNR) of the peaks from reactants and reaction products, with an overall standard deviation of chemical shift differences compared to reference spectra of 0.02ppm when measured on a 400MHz NMR spectrometer. Notably, the appearance of decoupling side-bands, which scale with peak intensity, appears to be of secondary importance.

  3. Hyperpolarization-Activated Currents and Subthreshold Resonance in Granule Cells of the Olfactory Bulb

    PubMed Central

    Hu, Ruilong; Ferguson, Katie A.; Meijer, Dimphna H.

    2016-01-01

    Abstract An important contribution to neural circuit oscillatory dynamics is the ongoing activation and inactivation of hyperpolarization-activated currents (Ih). Network synchrony dynamics play an important role in the initial processing of odor signals by the main olfactory bulb (MOB) and accessory olfactory bulb (AOB). In the mouse olfactory bulb, we show that Ih is present in granule cells (GCs), the most prominent inhibitory neuron in the olfactory bulb, and that Ih underlies subthreshold resonance in GCs. In accord with the properties of Ih, the currents exhibited sensitivity to changes in extracellular K+ concentration and ZD7288 (4-ethylphenylamino-1,2-dimethyl-6-methylaminopyrimidin chloride), a blocker of Ih. ZD7288 also caused GCs to hyperpolarize and increase their input resistance, suggesting that Ih is active at rest in GCs. The inclusion of cAMP in the intracellular solution shifted the activation of Ih to less negative potentials in the MOB, but not in the AOB, suggesting that channels with different subunit composition mediate Ih in these regions. Furthermore, we show that mature GCs exhibit Ih-dependent subthreshold resonance in the theta frequency range (4–12 Hz). Another inhibitory subtype in the MOB, the periglomerular cells, exhibited Ih-dependent subthreshold resonance in the delta range (1–4 Hz), while principal neurons, the mitral cells, do not exhibit Ih-dependent subthreshold resonance. Importantly, Ih size, as well as the strength and frequency of resonance in GCs, exhibited a postnatal developmental progression, suggesting that this development of Ih in GCs may differentially contribute to their integration of sensory input and contribution to oscillatory circuit dynamics. PMID:27844056

  4. Reaction monitoring using hyperpolarized NMR with scaling of heteronuclear couplings by optimal tracking

    NASA Astrophysics Data System (ADS)

    Zhang, Guannan; Schilling, Franz; Glaser, Steffen J.; Hilty, Christian

    2016-11-01

    Off-resonance decoupling using the method of Scaling of Heteronuclear Couplings by Optimal Tracking (SHOT) enables determination of heteronuclear correlations of chemical shifts in single scan NMR spectra. Through modulation of J-coupling evolution by shaped radio frequency pulses, off resonance decoupling using SHOT pulses causes a user-defined dependence of the observed J-splitting, such as the splitting of 13C peaks, on the chemical shift offset of coupled nuclei, such as 1H. Because a decoupling experiment requires only a single scan, this method is suitable for characterizing on-going chemical reactions using hyperpolarization by dissolution dynamic nuclear polarization (D-DNP). We demonstrate the calculation of [13C, 1H] chemical shift correlations of the carbanionic active sites from hyperpolarized styrene polymerized using sodium naphthalene as an initiator. While off resonance decoupling by SHOT pulses does not enhance the resolution in the same way as a 2D NMR spectrum would, the ability to obtain the correlations in single scans makes this method ideal for determination of chemical shifts in on-going reactions on the second time scale. In addition, we present a novel SHOT pulse that allows to scale J-splittings 50% larger than the respective J-coupling constant. This feature can be used to enhance the resolution of the indirectly detected chemical shift and reduce peak overlap, as demonstrated in a model reaction between p-anisaldehyde and isobutylamine. For both pulses, the accuracy is evaluated under changing signal-to-noise ratios (SNR) of the peaks from reactants and reaction products, with an overall standard deviation of chemical shift differences compared to reference spectra of 0.02 ppm when measured on a 400 MHz NMR spectrometer. Notably, the appearance of decoupling side-bands, which scale with peak intensity, appears to be of secondary importance.

  5. Endothelial-derived hyperpolarization contributes to acetylcholine-mediated vasodilation in human skin in a dose-dependent manner.

    PubMed

    Brunt, Vienna E; Fujii, Naoto; Minson, Christopher T

    2015-11-01

    Cutaneous acetylcholine (ACh)-mediated dilation is commonly used to assess microvascular function, but the mechanisms of dilation are poorly understood. Depending on dose and method of administration, nitric oxide (NO) and prostanoids are involved to varying extents and the roles of endothelial-derived hyperpolarizing factors (EDHFs) are unclear. In the present study, five incremental doses of ACh (0.01-100 mM) were delivered either as a 1-min bolus (protocol 1, n = 12) or as a ≥20-min continuous infusion (protocol 2, n = 10) via microdialysis fibers infused with 1) lactated Ringer, 2) tetraethylammonium (TEA) [a calcium-activated potassium channel (KCa) and EDHF inhibitor], 3) L-NNA+ketorolac [NO synthase (NOS) and cyclooxygenase (COX) inhibitors], and 4) TEA+L-NNA+Ketorolac. The hyperemic response was characterized as peak and area under the curve (AUC) cutaneous vascular conductance (CVC) for bolus infusions or plateau CVC for continuous infusions, and reported as %maximal CVC. In protocol 1, TEA, alone and combined with NOS+COX inhibition, attenuated peak CVC (100 mM Ringer 59 ± 6% vs. TEA 43 ± 5%, P < 0.05; L-NNA+ketorolac 35 ± 4% vs. TEA+L-NNA+ketorolac 25 ± 4%, P < 0.05) and AUC (Ringer 25,414 ± 3,528 vs. TEA 21,403 ± 3,416%·s, P < 0.05; L-NNA+ketorolac 25,628 ± 3,828%(.)s vs. TEA+L-NNA+ketorolac 20,772 ± 3,711%·s, P < 0.05), although these effects were only significant at the highest dose of ACh. At lower doses, TEA lengthened the total time of the hyperemic response (10 mM Ringer 609 ± 78 s vs. TEA 860 ± 67 s, P < 0.05). In protocol 2, TEA alone did not affect plateau CVC, but attenuated plateau in combination with NOS+COX inhibition (100 mM 50.4 ± 6.6% vs. 30.9 ± 6.3%, P < 0.05). Therefore, EDHFs contribute to cutaneous ACh-mediated dilation, but their relative contribution is altered by the dose and infusion procedure.

  6. Hyperpolarized [1,4-13C]-Diethylsuccinate: A Potential DNP Substrate for In Vivo Metabolic Imaging

    PubMed Central

    Billingsley, Kelvin L.; Josan, Sonal; Park, Jae Mo; Tee, Sui Seng; Spielman-Sun, Eleanor; Hurd, Ralph; Mayer, Dirk; Spielman, Daniel

    2014-01-01

    The tricarboxylic acid cycle (TCA) performs an essential role in the regulation of energy and metabolism, and deficiencies in this pathway are commonly correlated with various diseases. However, the development of non-invasive techniques for the assessment of the cycle in vivo has remained challenging. In this work, the applicability of a novel imaging agent, [1,4-13C]-diethylsuccinate, for hyperpolarized 13C metabolic imaging of the TCA cycle was explored. In vivo spectroscopic studies were conducted in conjunction with in vitro analyses to determine the metabolic fate of the imaging agent. Contrary to previous reports (Zacharias, N. M. et. al. J. Am. Chem. Soc. 2012, 134, 934-943), [13C]-labeled diethylsuccinate was primarily metabolized to succinate-derived products not originating from TCA cycle metabolism. These results illustrate potential issues of utilizing dialkyl ester analogs of TCA cycle intermediates as molecular probes for hyperpolarized 13C metabolic imaging. PMID:24421249

  7. Dipolar cross-relaxation modulates signal amplitudes in the 1H NMR spectrum of hyperpolarized [ 13C]formate

    NASA Astrophysics Data System (ADS)

    Merritt, Matthew E.; Harrison, Crystal; Mander, William; Malloy, Craig R.; Dean Sherry, A.

    2007-12-01

    The asymmetry in the doublet of a spin coupled to hyperpolarized 13C has been used previously to measure the initial polarization of 13C. We tested the hypothesis that a single observation of the 1H NMR spectrum of hyperpolarized 13C formate monitors 13C polarization. Depending on the microwave frequency during the polarization process, in-phase or out-of-phase doublets were observed in the 1H NMR spectrum. Even in this simple two-spin system, 13C polarization was not reflected in the relative area of the JCH doublet components due to strong heteronuclear cross-relaxation. The Solomon equations were used to model the proton signal as a function of time after polarization and to estimate 13C polarization from the 1H NMR spectra.

  8. Facilitated Anion Transport Induces Hyperpolarization of the Cell Membrane That Triggers Differentiation and Cell Death in Cancer Stem Cells.

    PubMed

    Soto-Cerrato, Vanessa; Manuel-Manresa, Pilar; Hernando, Elsa; Calabuig-Fariñas, Silvia; Martínez-Romero, Alicia; Fernández-Dueñas, Víctor; Sahlholm, Kristoffer; Knöpfel, Thomas; García-Valverde, María; Rodilla, Ananda M; Jantus-Lewintre, Eloisa; Farràs, Rosa; Ciruela, Francisco; Pérez-Tomás, Ricardo; Quesada, Roberto

    2015-12-23

    Facilitated anion transport potentially represents a powerful tool to modulate various cellular functions. However, research into the biological effects of small molecule anionophores is still at an early stage. Here we have used two potent anionophore molecules inspired in the structure of marine metabolites tambjamines to gain insight into the effect induced by these compounds at the cellular level. We show how active anionophores, capable of facilitating the transmembrane transport of chloride and bicarbonate in model phospholipid liposomes, induce acidification of the cytosol and hyperpolarization of plasma cell membranes. We demonstrate how this combined effect can be used against cancer stem cells (CSCs). Hyperpolarization of cell membrane induces cell differentiation and loss of stemness of CSCs leading to effective elimination of this cancer cell subpopulation.

  9. Fast volumetric imaging of ethanol metabolism in rat liver with hyperpolarized [1-13C]-pyruvate

    PubMed Central

    Josan, Sonal; Spielman, Daniel; Yen, Yi-Fen; Hurd, Ralph; Pfefferbaum, Adolf; Mayer, Dirk

    2012-01-01

    Rapid, volumetric imaging of hyperpolarized 13C compounds allows the real time measurement of metabolic activity and can be useful in distinguishing between normal and diseased tissues. This work extends a fast 2D under-sampled spiral magnetic resonance spectroscopic imaging (MRSI) sequence to provide volumetric coverage, acquiring a 16×16×12 matrix with a nominal 5 mm isotropic resolution in 4.5 s. The rapid acquisition enables a high temporal resolution for dynamic imaging. This dynamic 3D MRSI method was used to investigate hyperpolarized [1-13C]-pyruvate metabolism modulated by the administration of ethanol in rat liver. A significant increase in the pyruvate to lactate conversion was observed in the liver due to the greater availability of NADH from ethanol metabolism. PMID:22331837

  10. [Use of laser flow-type fluorescence aerosol particle counter to evaluate the concentration of microbes in the surface air under high dust content].

    PubMed

    Kalinin, Iu T; Vorob'ev, S A; Khramov, E N; Vorob'eva, E A; Kuznetsov, A P; Kiselev, O S

    2000-01-01

    The paper deals with the use of a laser flow-type fluorescence aerosol particle counter to evaluate the concentrations of microbes in the surface air under high dust content. Various circuits of flow-type optic aerosol recorders are analyzed. Flow spectral luminescence analysis of some particles flow while exciting the fourth harmonics of a pulse laser on yttrium-aluminium garnet with neodymium by ultraviolet radiation is shown to be the most optimum method for indication of individual aerosol particles. Experiments were conducted on the authors' model of a pilot plant based on this method. The model of a laser flow-type optic analyzer was developed for experimental studies that give a clear display of biological aerosols in complex aerosols. The laser flow-type analyzer-based unit developed may provide a fluorescence signal of aerosol particles in the flow of a sample and that light diffusion signal from them at an exciting light wavelength of 266 nm. Experiments with BVC aerosols and soil dust particles were conducted in different regions of Russia. They showed it possible to detect and to rapidly calculate soil microorganisms by laser flow-type fluorescence assay of individual particles when excited by ultraviolet radiation.

  11. XeNA: An automated ‘open-source’ 129Xe hyperpolarizer for clinical use

    PubMed Central

    Nikolaou, Panayiotis; Coffey, Aaron M.; Walkup, Laura L.; Gust, Brogan M.; Whiting, Nicholas; Newton, Hayley; Muradyan, Iga; Dabaghyan, Mikayel; Ranta, Kaili; Moroz, Gregory D.; Rosen, Matthew S.; Patz, Samuel; Barlow, Michael J.; Chekmenev, Eduard Y.; Goodson, Boyd M.

    2014-01-01

    Here we provide a full report on the construction, components, and capabilities of our consortium’s “open-source” large-scale (~1 L/hr) 129Xe hyperpolarizer for clinical, pre-clinical, and materials NMR/MRI (Nikolaou et al., Proc. Natl. Acad. Sci. USA, 110, 14150 (2013)). The ‘hyperpolarizer’ is automated and built mostly of off-the-shelf components; moreover, it is designed to be cost-effective and installed in both research laboratories and clinical settings with materials costing less than $125,000. The device runs in the xenon-rich regime (up to 1800 Torr Xe in 0.5 L) in either stopped-flow or single-batch mode—making cryo-collection of the hyperpolarized gas unnecessary for many applications. In-cell 129Xe nuclear spin polarization values of ~30-90% have been measured for Xe loadings of ~300-1600 Torr. Typical 129Xe polarization build-up and T1 relaxation time constants were ~8.5 min and ~1.9 hr respectively under our SEOP conditions; such ratios, combined with near-unity Rb electron spin polarizations enabled by the high resonant laser power (up to ~200 W), permits such high PXe values to be achieved despite the high in-cell Xe densities. Importantly, most of the polarization is maintained during efficient HP gas transfer to other containers, and ultra-long 129Xe relaxation times (up to nearly 6 hr) were observed in Tedlar bags following transport to a clinical 3 T scanner for MR spectroscopy and imaging as a prelude to in vivo experiments. The device has received FDA IND approval for a clinical study of COPD subjects. The primary focus of this paper is on the technical / engineering development of the polarizer, with the explicit goals of facilitating the adaptation of design features and operative modes into other laboratories, and of spurring the further advancement of HP-gas MR applications in biomedicine. PMID:24631715

  12. A novel slow hyperpolarization-activated potassium current (IK(SHA)) from a mouse hippocampal cell line.

    PubMed Central

    Wischmeyer, E; Karschin, A

    1997-01-01

    1. A slow hyperpolarization-activated inwardly rectifying K+ current (IK(SHA)) with novel characteristics was identified from the mouse embryonic hippocampus x neuroblastoma cell line HN9.10e. 2. The non-inactivating current activated negative to a membrane potential of -80 mV with slow and complex activation kinetics (tau act approximately 1-7 s) and a characteristic delay of 1-10 s (-80 to -140 mV) that was linearly dependent on the membrane potential. 3. Tail currents and instantaneous open channel currents determined through fast voltage ramps reversed at the K+ equilibrium potential (EK) indicating that primarily K+, but not Na+, permeated the channels. 4. IK(SHA) was unaffected by altering the intracellular Ca2+ concentration between approximately 0 and 10 microM, but was susceptible to block by 5 mM extracellular Ca2+, Ba2+ (Ki = 0.42 mM), and Cs+ (Ki = 2.77 mM) 5. In cells stably transformed with M2 muscarinic receptors, IK(SHA) was rapidly, but reversibly, suppressed by application of micromolar concentrations of muscarine. 6. At the single channel level K(SHA) channel openings were observed with the characteristic delay upon membrane hyperpolarization. Analysis of unitary currents revealed an inwardly rectifying I-V profile and a channel slope conductance of 7 pS. Channel activity persisted in the inside-out configuration for many minutes. 7. It is concluded that IK(SHA) in HN9.10e cells represents a novel K+ current, which is activated upon membrane hyperpolarization. It is functionally different from both classic inwardly rectifying IKir currents and other cationic hyperpolarization-activated IH currents that have been previously described in neuronal or glial cells. Images Figure 1 Figure 2 Figure 6 Figure 7 Figure 8 PMID:9401967

  13. HYPERPOLARIZED 13C MAGNETIC RESONANCE AND ITS USE IN METABOLIC ASSESSMENT OF CULTURED CELLS AND PERFUSED ORGANS

    PubMed Central

    Lumata, Lloyd; Yang, Chendong; Ragavan, Mukundan; Carpenter, Nicholas; DeBerardinis, Ralph J.; Merritt, Matthew E.

    2016-01-01

    Diseased tissue is often characterized by abnormalities in intermediary metabolism. Observing these alterations in situ may lead to an improved understanding of pathological processes and novel ways to monitor these processes non-invasively in human patients. Although 13C is a stable isotope safe for use in animal models of disease as well as human subjects, its utility as a metabolic tracer has largely been limited to ex vivo analyses employing analytical techniques like mass spectrometry or nuclear magnetic resonance spectroscopy. Neither of these techniques is suitable for non-invasive metabolic monitoring, and the low abundance and poor gyromagnetic ratio of conventional 13C make it a poor nucleus for imaging. However, the recent advent of hyperpolarization methods, particularly dynamic nuclear polarization (DNP), make it possible to enhance the spin polarization state of 13C by many orders of magnitude, resulting in a temporary amplification of the signal sufficient for monitoring kinetics of enzyme-catalyzed reactions in living tissue through magnetic resonance spectroscopy or magnetic resonance imaging. Here we review DNP techniques to monitor metabolism in cultured cells, perfused hearts, and perfused livers, focusing on our experiences with hyperpolarized [1-13C]pyruvate. We present detailed approaches to optimize the DNP procedure, streamline biological sample preparation, and maximize detection of specific metabolic activities. We also discuss practical aspects in the choice of metabolic substrates for hyperpolarization studies, and outline some of the current technical and conceptual challenges in the field, including efforts to use hyperpolarization to quantify metabolic rates in vivo. PMID:26358902

  14. Deuteration of a molecular probe for DNP hyperpolarization--a new approach and validation for choline chloride.

    PubMed

    Allouche-Arnon, Hyla; Lerche, Mathilde H; Karlsson, Magnus; Lenkinski, Robert E; Katz-Brull, Rachel

    2011-01-01

    The promising dynamic nuclear polarization (DNP) for hyperpolarized (13)C-MRI/MRS of real-time metabolism in vivo is challenged by the limited number of agents with the required physical and biological properties. The physical requirement of a liquid-state T(1) of tens of seconds is mostly found for (13)C-carbons in small molecules that have no direct protons attached, i.e. carbonyl, carboxyl and certain quaternary carbons. Unfortunately, such carbon positions do not exist in a large number of metabolic agents, and chemical shift dispersion often limits detection of their chemical evolution. We have previously shown that direct deuteration of protonated carbon positions significantly prolongs the (13)C T(1) in the liquid state and provides potential (13)C-labeled agents with differential chemical shift with respect to metabolism. The Choline Molecular Probe [1,1,2,2-D(4), 2-(13)C]choline chloride (CMP2) has recently been introduced as a means of studying choline metabolism in a hyperpolarized state. Here, the biophysical properties of CMP2 were characterized and compared with those of [1-(13)C]pyruvate to evaluate the impact of molecular probe deuteration. The CMP2 solid-state polarization build-up time constant (30 min) and polarization level (24%) were comparable to those of [1-(13)C]pyruvate. Both compounds' liquid state T(1) increased with temperature. The high-field T(1) of CMP2 compared favorably with [1-(13)C]pyruvate. Thus, a deuterated agent demonstrated physical properties comparable to a hyperpolarized compound of already proven value, whereas both showed chemical shift dispersion that allowed monitoring of their metabolism. It is expected that the use of deuterated carbon-13 positions as reporting hyperpolarized nuclei will substantially expand the library of agents for DNP-MR.

  15. Hyperpolarization of the Membrane Potential Caused by Somatostatin in Dissociated Human Pituitary Adenoma Cells that Secrete Growth Hormone

    NASA Astrophysics Data System (ADS)

    Yamashita, Naohide; Shibuya, Naohiko; Ogata, Etsuro

    1986-08-01

    Membrane electrical properties and the response to somatostatin were examined in dissociated human pituitary adenoma cells that secrete growth hormone (GH). Under current clamp condition with a patch electrode, the resting potential was -52.4 ± 8.0 mV, and spontaneous action potentials were observed in 58% of the cells. Under voltage clamp condition an outward K+ current, a tetrodotoxin-sensitive Na+ current, and a Ca2+ current were observed. Cobalt ions suppressed the Ca2+ current. The threshold of Ca2+ current activation was about -60 mV. Somatostatin elicited a membrane hyperpolarization associated with increased membrane permeability in these cells. The reversal potential of somatostatin-induced hyperpolarization was -78.4 ± 4.3 mV in 6 mM K+ medium and -97.2 ± 6.4 mV in 3 mM K+ medium. These reversal potential values and a shift with the external K+ concentration indicated that membrane hyperpolarization was caused by increased permeability to K+. The hyperpolarized membrane potential induced by somatostatin was -63.6 ± 5.9 mV in the standard medium. This level was subthreshold for Ca2+ and Na+ currents and was sufficient to inhibit spontaneous action potentials. Hormone secretion was significantly suppressed by somatostatin and cobalt ions. Therefore, we suggest that Ca2+ entering the cell through voltage-dependent channels are playing an important role for GH secretion and that somatostatin suppresses GH secretion by blocking Ca2+ currents. Finally, we discuss other possibilities for the inhibitory effect of somatostatin on GH secretion.

  16. Continuous flow Overhauser dynamic nuclear polarization of water in the fringe field of a clinical magnetic resonance imaging system for authentic image contrast

    PubMed Central

    Lingwood, Mark D.; Siaw, Ting Ann; Sailasuta, Napapon; Ross, Brian D.; Bhattacharya, Pratip; Han, Songi

    2016-01-01

    We describe and demonstrate a system to generate hyperpolarized water in the 0.35 T fringe field of a clinical 1.5 T whole-body magnetic resonance imaging (MRI) magnet. Once generated, the hyperpolarized water is quickly and continuously transferred from the 0.35 T fringe to the 1.5 T center field of the same magnet for image acquisition using standard MRI equipment. The hyperpolarization is based on Overhauser dynamic nuclear polarization (DNP), which effectively and quickly transfers the higher spin polarization of free radicals to nuclear spins at ambient temperatures. We visualize the dispersion of hyperpolarized water as it flows through water-saturated systems by utilizing an observed −15 fold DNP signal enhancement with respect to the unenhanced 1H MRI signal of water at 1.5 T. The experimental DNP apparatus presented here is readily portable and can be brought to and used with any conventional unshielded MRI system. A new method of immobilizing radicals to gel beads via polyelectrolyte linker arms is described, which led to superior flow Overhauser DNP performance compared to previously presented gels. We discuss the general applicability of Overhauser DNP hyperpolarization of water and aqueous solutions in the fringe field of commercially available magnets with central fields up to 4.7 Tesla. PMID:20541445

  17. Presynaptic hyperpolarization induces a fast analogue modulation of spike-evoked transmission mediated by axonal sodium channels

    PubMed Central

    Rama, Sylvain; Zbili, Mickaël; Bialowas, Andrzej; Fronzaroli-Molinieres, Laure; Ankri, Norbert; Carlier, Edmond; Marra, Vincenzo; Debanne, Dominique

    2015-01-01

    In the mammalian brain, synaptic transmission usually depends on presynaptic action potentials (APs) in an all-or-none (or digital) manner. Recent studies suggest, however, that subthreshold depolarization in the presynaptic cell facilitates spike-evoked transmission, thus creating an analogue modulation of a digital process (or analogue–digital (AD) modulation). At most synapses, this process is slow and not ideally suited for the fast dynamics of neural networks. We show here that transmission at CA3–CA3 and L5–L5 synapses can be enhanced by brief presynaptic hyperpolarization such as an inhibitory postsynaptic potential (IPSP). Using dual soma–axon patch recordings and live imaging, we find that this hyperpolarization-induced AD facilitation (h-ADF) is due to the recovery from inactivation of Nav channels controlling AP amplitude in the axon. Incorporated in a network model, h-ADF promotes both pyramidal cell synchrony and gamma oscillations. In conclusion, cortical excitatory synapses in local circuits display hyperpolarization-induced facilitation of spike-evoked synaptic transmission that promotes network synchrony. PMID:26657943

  18. Hydrogen sulfide induces hyperpolarization and decreases the exocytosis of secretory granules of rat GH3 pituitary tumor cells.

    PubMed

    Mustafina, Alsu N; Yakovlev, Aleksey V; Gaifullina, Aisylu Sh; Weiger, Thomas M; Hermann, Anton; Sitdikova, Guzel F

    2015-10-02

    The aim of the present study was to evaluate the effects of hydrogen sulfide (H2S) on the membrane potential, action potential discharge and exocytosis of secretory granules in neurosecretory pituitary tumor cells (GH3). The H2S donor - sodium hydrosulfide (NaHS) induced membrane hyperpolarization, followed by truncation of spontaneous electrical activity and decrease of the membrane resistance. The NaHS effect was dose-dependent with an EC50 of 152 μM (equals effective H2S of 16-19 μM). NaHS effects were not altered after inhibition of maxi conductance calcium-activated potassium (BK) channels by tetraethylammonium or paxilline, but were significantly reduced after inhibition or activation of ATP-dependent potassium channels (KATP) by glibenclamide or by diazoxide, respectively. In whole-cell recordings NaHS increased the amplitude of KATP currents, induced by hyperpolarizing pulses and subsequent application of glibenclamide decreased currents to control levels. Using the fluorescent dye FM 1-43 exocytosis of secretory granules was analyzed in basal and stimulated conditions (high K(+) external solution). Prior application of NaHS decreased the fluorescence of the cell membrane in both conditions which links with activation of KATP currents (basal secretion) and activation of KATP currents and BK-currents (stimulated exocytosis). We suggest that H2S induces hyperpolarization of GH3 cells by activation of KATP channels which results in a truncation of spontaneous action potentials and a decrease of hormone release.

  19. Multidimensional Mapping of Spin-Exchange Optical Pumping in Clinical-Scale Batch-Mode 129Xe Hyperpolarizers

    PubMed Central

    2015-01-01

    We present a systematic, multiparameter study of Rb/129Xe spin-exchange optical pumping (SEOP) in the regimes of high xenon pressure and photon flux using a 3D-printed, clinical-scale stopped-flow hyperpolarizer. In situ NMR detection was used to study the dynamics of 129Xe polarization as a function of SEOP-cell operating temperature, photon flux, and xenon partial pressure to maximize 129Xe polarization (PXe). PXe values of 95 ± 9%, 73 ± 4%, 60 ± 2%, 41 ± 1%, and 31 ± 1% at 275, 515, 1000, 1500, and 2000 Torr Xe partial pressure were achieved. These PXe polarization values were separately validated by ejecting the hyperpolarized 129Xe gas and performing low-field MRI at 47.5 mT. It is shown that PXe in this high-pressure regime can be increased beyond already record levels with higher photon flux and better SEOP thermal management, as well as optimization of the polarization dynamics, pointing the way to further improvements in hyperpolarized 129Xe production efficiency. PMID:24731261

  20. Reproducibility study for free-breathing measurements of pyruvate metabolism using hyperpolarized (13) C in the heart.

    PubMed

    Lau, Angus Z; Chen, Albert P; Barry, Jennifer; Graham, John J; Dominguez-Viqueira, William; Ghugre, Nilesh R; Wright, Graham A; Cunningham, Charles H

    2013-04-01

    Spatially resolved images of hyperpolarized (13) C substrates and their downstream products provide insight into real-time metabolic processes occurring in vivo. Recently, hyperpolarized (13) C pyruvate has been used to characterize in vivo cardiac metabolism in the rat and pig, but accurate and reproducible measurements remain challenging due to the limited period available for imaging as well as physiological motion. In this article, time-resolved cardiac- and respiratory-gated images of [1-(13) C] pyruvate, [1-(13) C] lactate, and (13) C bicarbonate in the heart are acquired without the need for a breathhold. The robustness of these free-breathing measurements is demonstrated using the time-resolved data to produce a normalized metric of pyruvate dehydrogenase and lactate dehydrogenase activity in the heart. The values obtained are reproducible in a controlled metabolic state. In a 60-min ischemia/reperfusion model, significant differences in hyperpolarized bicarbonate and lactate, normalized using the left ventricular pyruvate signal, were detected between scans performed at baseline and 45 min after reperfusion. The sequence is anticipated to improve quantitative measurements of cardiac metabolism, leading to feasible validation studies using fewer subjects, and potentially improved diagnosis, serial monitoring, and treatment of cardiac disease in patients.

  1. Hyperpolarized 13C allows a direct measure of flux through a single enzyme-catalyzed step by NMR

    PubMed Central

    Merritt, Matthew E.; Harrison, Crystal; Storey, Charles; Jeffrey, F. Mark; Sherry, A. Dean; Malloy, Craig R.

    2007-01-01

    13C NMR is a powerful tool for monitoring metabolic fluxes in vivo. The recent availability of automated dynamic nuclear polarization equipment for hyperpolarizing 13C nuclei now offers the potential to measure metabolic fluxes through select enzyme-catalyzed steps with substantially improved sensitivity. Here, we investigated the metabolism of hyperpolarized [1-13C1]pyruvate in a widely used model for physiology and pharmacology, the perfused rat heart. Dissolved 13CO2, the immediate product of the first step of the reaction catalyzed by pyruvate dehydrogenase, was observed with a temporal resolution of ≈1 s along with H13CO3−, the hydrated form of 13CO2 generated catalytically by carbonic anhydrase. In hearts presented with the medium-chain fatty acid octanoate in addition to hyperpolarized [1-13C1]pyruvate, production of 13CO2 and H13CO3− was suppressed by ≈90%, whereas the signal from [1-13C1]lactate was enhanced. In separate experiments, it was shown that O2 consumption and tricarboxylic acid (TCA) cycle flux were unchanged in the presence of added octanoate. Thus, the rate of appearance of 13CO2 and H13CO3− from [1-13C1]pyruvate does not reflect production of CO2 in the TCA cycle but rather reflects flux through pyruvate dehydrogenase exclusively. PMID:18056642

  2. Multidimensional mapping of spin-exchange optical pumping in clinical-scale batch-mode 129Xe hyperpolarizers.

    PubMed

    Nikolaou, Panayiotis; Coffey, Aaron M; Ranta, Kaili; Walkup, Laura L; Gust, Brogan M; Barlow, Michael J; Rosen, Matthew S; Goodson, Boyd M; Chekmenev, Eduard Y

    2014-05-08

    We present a systematic, multiparameter study of Rb/(129)Xe spin-exchange optical pumping (SEOP) in the regimes of high xenon pressure and photon flux using a 3D-printed, clinical-scale stopped-flow hyperpolarizer. In situ NMR detection was used to study the dynamics of (129)Xe polarization as a function of SEOP-cell operating temperature, photon flux, and xenon partial pressure to maximize (129)Xe polarization (PXe). PXe values of 95 ± 9%, 73 ± 4%, 60 ± 2%, 41 ± 1%, and 31 ± 1% at 275, 515, 1000, 1500, and 2000 Torr Xe partial pressure were achieved. These PXe polarization values were separately validated by ejecting the hyperpolarized (129)Xe gas and performing low-field MRI at 47.5 mT. It is shown that PXe in this high-pressure regime can be increased beyond already record levels with higher photon flux and better SEOP thermal management, as well as optimization of the polarization dynamics, pointing the way to further improvements in hyperpolarized (129)Xe production efficiency.

  3. Fast Determination of Flip Angle and T1 in Hyperpolarized Gas MRI During a Single Breath-Hold

    PubMed Central

    Zhong, Jianping; Ruan, Weiwei; Han, Yeqing; Sun, Xianping; Ye, Chaohui; Zhou, Xin

    2016-01-01

    MRI of hyperpolarized media, such as 129Xe and 3He, shows great potential for clinical applications. The optimal use of the available spin polarization requires accurate flip angle calibrations and T1 measurements. Traditional flip angle calibration methods are time-consuming and suffer from polarization losses during T1 relaxation. In this paper, we propose a method to simultaneously calibrate flip angles and measure T1 in vivo during a breath-hold time of less than 4 seconds. We demonstrate the accuracy, robustness and repeatability of this method and contrast it with traditional methods. By measuring the T1 of hyperpolarized gas, the oxygen pressure in vivo can be calibrated during the same breath hold. The results of the calibration have been applied in variable flip angle (VFA) scheme to obtain a stable steady-state transverse magnetization. Coupled with this method, the ultra-short TE (UTE) and constant VFA (CVFA) schemes are expected to give rise to new applications of hyperpolarized media. PMID:27169670

  4. Endothelium-derived hyperpolarization and coronary vasodilation: diverse and integrated roles of epoxyeicosatrienoic acids, hydrogen peroxide and gap junctions

    PubMed Central

    Ellinsworth, David C.; Sandow, Shaun L.; Shukla, Nilima; Liu, Yanping; Jeremy, Jamie Y.; Gutterman, David D.

    2015-01-01

    Myocardial perfusion and coronary vascular resistance are regulated by signalling metabolites released from the local myocardium that act either directly on the vascular smooth muscle cells (VSMC) or indirectly via stimulation of the endothelium. A prominent mechanism of vasodilation is endothelium-derived hyperpolarization (EDH) of the arteriolar smooth muscle, with epoxyeicosatrienoic acids (EETs) and hydrogen peroxide (H2O2) playing important roles in EDH in the coronary microcirculation. In some cases, EETs and H2O2 are released as transferable hyperpolarizing factors (EDHFs) that act directly on the VSMCs. By contrast, EETs and H2O2 can also promote endothelial Ca2+-activated K+ channel activity secondary to the amplification of extracellular Ca2+ influx and Ca2+ mobilization from intracellular stores, respectively. The resulting endothelial hyperpolarization may subsequently conduct to the media via myoendothelial gap junctions, or potentially lead to the release of a chemically-distinct factor(s). Furthermore, in human isolated coronary arterioles dilator signalling involving EETs and H2O2 may be integrated; being either complimentary or inhibitory depending on the stimulus. With an emphasis on the human coronary microcirculation, this review addresses the diverse and integrated mechanisms by which EETs and H2O2 regulate vessel tone, and also examines the hypothesis that myoendothelial microdomain signalling facilitates EDH activity in the human heart. PMID:26541094

  5. Gas cells for 3He hyperpolarized via spin-exchange optical pumping

    NASA Astrophysics Data System (ADS)

    Kim, W.; Stepanyan, S. S.; Kim, A.; Tan, J. A.; Woo, S.

    2016-01-01

    We present a device for the production of hyperpolarized 3He, which is widely used in spinrelated nuclear physics research. Spin-exchange optical pumping (SEOP) is employed to polarize 3He enclosed in a circular borosilicate glass cell suitable not only for the production of polarized gas but also for its storage. The portable glass cell can, thus, be transported to any other research facility. The glass cell can be refilled several times. Special attention is given to the preparation and the filling of the cell to minimize the impurities on its walls and in the gas. We employ glass tubes with shorter lengths and larger diameters in the gas-filling system to achieve the improvement in the air flow necessary to obtain purer polarized 3He samples. The cell is prepared, and after it has been filled with rubidium (Rb) and 3He-N2 mixture, it is sealed under high vacuum conditions. The cell containing the mixture is exposed to circularly-polarized laser light with a wavelength of 795 nm at temperatures of 180 - 220 °C for SEOP. The polarization of 3He is measured via nuclear magnetic resonance (NMR). We obtained 40% polarized 3He in less than 15 hours and 50% in about 25 hours. The longitudinal relaxation time T 1 of the polarized 3He we measured was about 58 hours.

  6. Hyperpolarized 83Kr magnetic resonance imaging of alveolar degradation in a rat model of emphysema

    PubMed Central

    Lilburn, David M. L.; Lesbats, Clémentine; Six, Joseph S.; Dubuis, Eric; Yew-Booth, Liang; Shaw, Dominick E.; Belvisi, Maria G.; Birrell, Mark A.; Pavlovskaya, Galina E.; Meersmann, Thomas

    2015-01-01

    Hyperpolarized 83Kr surface quadrupolar relaxation (SQUARE) generates MRI contrast that was previously shown to correlate with surface-to-volume ratios in porous model surface systems. The underlying physics of SQUARE contrast is conceptually different from any other current MRI methodology as the method uses the nuclear electric properties of the spin I = 9/2 isotope 83Kr. To explore the usage of this non-radioactive isotope for pulmonary pathophysiology, MRI SQUARE contrast was acquired in excised rat lungs obtained from an elastase-induced model of emphysema. A significant 83Kr T1 relaxation time increase in the SQUARE contrast was found in the elastase-treated lungs compared with the baseline data from control lungs. The SQUARE contrast suggests a reduction in pulmonary surface-to-volume ratio in the emphysema model that was validated by histology. The finding supports usage of 83Kr SQUARE as a new biomarker for surface-to-volume ratio changes in emphysema. PMID:25994296

  7. Hyperpolarization-Activated Current Induces Period-Doubling Cascades and Chaos in a Cold Thermoreceptor Model.

    PubMed

    Xu, Kesheng; Maidana, Jean P; Caviedes, Mauricio; Quero, Daniel; Aguirre, Pablo; Orio, Patricio

    2017-01-01

    In this article, we describe and analyze the chaotic behavior of a conductance-based neuronal bursting model. This is a model with a reduced number of variables, yet it retains biophysical plausibility. Inspired by the activity of cold thermoreceptors, the model contains a persistent Sodium current, a Calcium-activated Potassium current and a hyperpolarization-activated current (Ih) that drive a slow subthreshold oscillation. Driven by this oscillation, a fast subsystem (fast Sodium and Potassium currents) fires action potentials in a periodic fashion. Depending on the parameters, this model can generate a variety of firing patterns that includes bursting, regular tonic and polymodal firing. Here we show that the transitions between different firing patterns are often accompanied by a range of chaotic firing, as suggested by an irregular, non-periodic firing pattern. To confirm this, we measure the maximum Lyapunov exponent of the voltage trajectories, and the Lyapunov exponent and Lempel-Ziv's complexity of the ISI time series. The four-variable slow system (without spiking) also generates chaotic behavior, and bifurcation analysis shows that this is often originated by period doubling cascades. Either with or without spikes, chaos is no longer generated when the Ih is removed from the system. As the model is biologically plausible with biophysically meaningful parameters, we propose it as a useful tool to understand chaotic dynamics in neurons.

  8. Dopamine and baclofen inhibit the hyperpolarization-activated cation current in rat ventral tegmental neurones.

    PubMed Central

    Jiang, Z G; Pessia, M; North, R A

    1993-01-01

    1. Whole-cell patch-clamp recordings were made from dopamine-containing ventral tegmental area neurones in slices of rat midbrain. An inward current (Ih) was activated by hyperpolarization from -60 mV. 2. Dopamine (30 microM) reduced the amplitude of Ih by 10-30% at potentials from -70 to -120 mV. The effect was concentration dependent, mimicked by the D2 agonist quinpirole, and prevented by the D2 antagonist (-)-sulpiride. Baclofen (0.3-3 microM) also inhibited Ih; this action was antagonized by 2-hydroxysaclofen but not by (-)-sulpiride. The decrease in Ih resulted from a reduction in the maximal current with no change in the voltage dependence. 3. The action of dopamine was unaffected by cadmium (200 microM), forskolin (10 microM), the adenylyl cyclase inhibitor 2',3'-dideoxyadenosine (100 microM), or by intracellular solution containing cyclic AMP (2 mM). 4. Ih was progressively reduced during the first 5-10 min of recording with electrodes containing guanosine 5'-O-(3-thiotriphosphate); after this time, dopamine had no further effect. 5. It is concluded that agonists acting at D2 receptors and GABAB receptors reduce Ih in ventral tegmental neurones. PMID:8392580

  9. Effects of Diffusion Time on Short-Range Hyperpolarized 3He Diffusivity Measurements in Emphysema

    PubMed Central

    Gierada, David S.; Woods, Jason C.; Bierhals, Andrew J.; Bartel, Seth T.; Ritter, Jon H.; Choong, Cliff K.; Das, Nitin A.; Hong, Cheng; Pilgram, Thomas K.; Chang, Yulin V.; Jacob, Richard E.; Hogg, James C.; Battafarano, Richard J.; Cooper, Joel D.; Meyers, Bryan F.; Patterson, G. Alexander; Yablonskiy, Dmitriy A.; Conradi, Mark S.

    2010-01-01

    Purpose To characterize the effect of diffusion time on short-range hyperpolarized 3He MR diffusion measurements across a wide range of emphysema severity. Materials and Methods 3He diffusion MR imaging was performed on 19 lungs or lobes resected from 18 subjects with varying degrees of emphysema using 3 diffusion times (1.6 msec, 5 msec, and 10 msec) at constant b value. Emphysema severity was quantified as the mean apparent diffusion coefficient (ADC) and as the percentage of pixels with ADC higher than multiple thresholds from 0.30–0.55 cm2/sec (ADC index). Quantitative histology (mean linear intercept) was obtained in 10 of the lung specimens from 10 of the subjects. Results The mean ADCs with diffusion times of 1.6, 5.0, and 10.0 msec were 0.46, 0.40, and 0.37 cm2/sec, respectively (P <0.0001, ANOVA). There was no relationship between the ADC magnitude and the effect of diffusion time on ADC values. Mean linear intercept correlated with ADC (r=0.91–0.94, P<0.001) and ADC index (r=0.78–0.92, P<0.01) at all diffusion times. Conclusion Decreases in ADC with longer diffusion time were unrelated to emphysema severity. The strong correlations between the ADC at all diffusion times tested and quantitative histology demonstrate that the ADC is a robust measure of emphysema. PMID:19787725

  10. Hyperpolarized 3He functional magnetic resonance imaging of bronchoscopic airway bypass in chronic obstructive pulmonary disease

    PubMed Central

    Mathew, Lindsay; Kirby, Miranda; Farquhar, Donald; Licskai, Christopher; Santyr, Giles; Etemad-Rezai, Roya; Parraga, Grace; McCormack, David G

    2012-01-01

    A 73-year-old exsmoker with Global initiative for chronic Obstructive Lung Disease stage III chronic obstructive pulmonary disease underwent airway bypass (AB) as part of the Exhale Airway Stents for Emphysema (EASE) trial, and was the only EASE subject to undergo hyperpolarized 3He magnetic resonance imaging for evaluation of lung function pre- and post-AB. 3He magnetic resonance imaging was acquired twice previously (32 and eight months pre-AB) and twice post-AB (six and 12 months post-AB). Six months post-AB, his increase in forced vital capacity was <12% predicted, and he was classified as an AB nonresponder. However, post-AB, he also demonstrated improvements in quality of life scores, 6 min walk distance and improvements in 3He gas distribution in the regions of stent placement. Given the complex relationship between well-established pulmonary function and quality of life measurements, the present case provides evidence of the value-added information functional imaging may provide in chronic obstructive pulmonary disease interventional studies. PMID:22332133

  11. Hyperpolarized 3He functional magnetic resonance imaging of bronchoscopic airway bypass in chronic obstructive pulmonary disease.

    PubMed

    Mathew, Lindsay; Kirby, Miranda; Farquhar, Donald; Licskai, Christopher; Santyr, Giles; Etemad-Rezai, Roya; Parraga, Grace; McCormack, David G

    2012-01-01

    A 73-year-old exsmoker with Global initiative for chronic Obstructive Lung Disease stage III chronic obstructive pulmonary disease underwent airway bypass (AB) as part of the Exhale Airway Stents for Emphysema (EASE) trial, and was the only EASE subject to undergo hyperpolarized 3He magnetic resonance imaging for evaluation of lung function pre- and post-AB. 3He magnetic resonance imaging was acquired twice previously (32 and eight months pre-AB) and twice post-AB (six and 12 months post-AB). Six months post-AB, his increase in forced vital capacity was <12% predicted, and he was classified as an AB nonresponder. However, post-AB, he also demonstrated improvements in quality of life scores, 6 min walk distance and improvements in 3He gas distribution in the regions of stent placement. Given the complex relationship between well-established pulmonary function and quality of life measurements, the present case provides evidence of the value-added information functional imaging may provide in chronic obstructive pulmonary disease interventional studies.

  12. Essential Role of CFTR in PKA-Dependent Phosphorylation, Alkalinization, and Hyperpolarization During Human Sperm Capacitation.

    PubMed

    Puga Molina, Lis C; Pinto, Nicolás A; Torres Rodríguez, Paulina; Romarowski, Ana; Vicens Sanchez, Alberto; Visconti, Pablo E; Darszon, Alberto; Treviño, Claudia L; Buffone, Mariano G

    2017-06-01

    Mammalian sperm require to spend a limited period of time in the female reproductive tract to become competent to fertilize in a process called capacitation. It is well established that HCO3(-) is essential for capacitation because it activates the atypical soluble adenylate cyclase ADCY10 leading to cAMP production, and promotes alkalinization of cytoplasm, and membrane hyperpolarization. However, how HCO3(-) is transported into the sperm is not well understood. There is evidence that CFTR activity is involved in the human sperm capacitation but how this channel is integrated in the complex signaling cascades associated with this process remains largely unknown. In the present work, we have analyzed the extent to which CFTR regulates different events in human sperm capacitation. We observed that inhibition of CFTR affects HCO3(-) -entrance dependent events resulting in lower PKA activity. CFTR inhibition also affected cAMP/PKA-downstream events such as the increase in tyrosine phosphorylation, hyperactivated motility, and acrosome reaction. In addition, we demonstrated for the first time, that CFTR and PKA activity are essential for the regulation of intracellular pH, and membrane potential in human sperm. Addition of permeable cAMP partially recovered all the PKA-dependent events altered in the presence of inh-172 which is consistent with a role of CFTR upstream of PKA activation. J. Cell. Physiol. 232: 1404-1414, 2017. © 2016 Wiley Periodicals, Inc.

  13. Contribution of endothelium-derived hyperpolarizing factor to exercise-induced vasodilation in health and hypercholesterolemia.

    PubMed

    Ozkor, Muhiddin A; Hayek, Salim S; Rahman, Ayaz M; Murrow, Jonathan R; Kavtaradze, Nino; Lin, Ji; Manatunga, Amita; Quyyumi, Arshed A

    2015-02-01

    The role of endothelium-derived hyperpolarizing factor (EDHF) in either the healthy circulation or in those with hypercholesterolemia is unknown. In healthy and hypercholesterolemic subjects, we measured forearm blood flow (FBF) using strain-gauge plethysmography at rest, during graded handgrip exercise, and after sodium nitroprusside infusion. Measurements were repeated after l-NMMA, tetraethylammonium (TEA), and combined infusions. At rest, l-NMMA infusion reduced FBF in healthy but not hypercholesterolemic subjects. At peak exercise, vasodilation was lower in hypercholesterolemic compared to healthy subjects (274% vs 438% increase in FBF, p=0.017). TEA infusion reduced exercise-induced vasodilation in both healthy and hypercholesterolemic subjects (27%, p<0.0001 and -20%, p<0.0001, respectively). The addition of l-NMMA to TEA further reduced FBF in healthy (-14%, p=0.012) but not in hypercholesterolemic subjects, indicating a reduced nitric oxide and greater EDHF-mediated contribution to exercise-induced vasodilation in hypercholesterolemia. In conclusion, exercise-induced vasodilation is impaired and predominantly mediated by EDHF in hypercholesterolemic subjects. CLINICAL TRIAL REGISTRATION IDENTIFIER NCT00166166:

  14. Mitochondria Targeted and Intracellular Biothiol Triggered Hyperpolarized (129)Xe Magnetofluorescent Biosensor.

    PubMed

    Zeng, Qingbin; Guo, Qianni; Yuan, Yaping; Yang, Yuqi; Zhang, Bin; Ren, Lili; Zhang, Xiaoxiao; Luo, Qing; Liu, Maili; Bouchard, Louis-S; Zhou, Xin

    2017-02-21

    Biothiols such as gluthathione (GSH), cysteine (Cys), homocysteine (Hcy), and thioredoxin (Trx) play vital roles in cellular metabolism. Various diseases are associated with abnormal cellular biothiol levels. Thus, the intracellular detection of biothiol levels could be a useful diagnostic tool. A number of methods have been developed to detect intracellular thiols, but sensitivity and specificity problems have limited their applications. To address these limitations, we have designed a new biosensor based on hyperpolarized xenon magnetic resonance detection, which can be used to detect biothiol levels noninvasively. The biosensor is a multimodal probe that incorporates a cryptophane-A cage as (129)Xe NMR reporter, a naphthalimide moiety as fluorescence reporter, a disulfide bond as thiol-specific cleavable group, and a triphenylphosphonium moiety as mitochondria targeting unit. When the biosensor interacts with biothiols, disulfide bond cleavage leads to enhancements in the fluorescence intensity and changes in the (129)Xe chemical shift. Using Hyper-CEST (chemical exchange saturation transfer) NMR, our biosensor shows a low detection limit at picomolar (10(-10) M) concentration, which makes a promise to detect thiols in cells. The biosensor can detect biothiol effectively in live cells and shows good targeting ability to the mitochondria. This new approach not only offers a practical technique to detect thiols in live cells, but may also present an excellent in vivo test platform for xenon biosensors.

  15. Optimizing Flip Angles for Metabolic Rate Estimation in Hyperpolarized Carbon-13 MRI.

    PubMed

    Maidens, John; Gordon, Jeremy W; Arcak, Murat; Larson, Peder E Z

    2016-11-01

    Hyperpolarized carbon-13 magnetic resonance imaging has enabled the real-time observation of perfusion and metabolism in vivo. These experiments typically aim to distinguish between healthy and diseased tissues based on the rate at which they metabolize an injected substrate. However, existing approaches to optimizing flip angle sequences for these experiments have focused on indirect metrics of the reliability of metabolic rate estimates, such as signal variation and signal-to-noise ratio. In this paper we present an optimization procedure that focuses on maximizing the Fisher information about the metabolic rate. We demonstrate through numerical simulation experiments that flip angles optimized based on the Fisher information lead to lower variance in metabolic rate estimates than previous flip angle sequences. In particular, we demonstrate a 20% decrease in metabolic rate uncertainty when compared with the best competing sequence. We then demonstrate appropriateness of the mathematical model used in the simulation experiments with in vivo experiments in a prostate cancer mouse model. While there is no ground truth against which to compare the parameter estimates generated in the in vivo experiments, we demonstrate that our model used can reproduce consistent parameter estimates for a number of flip angle sequences.

  16. Hyperpolarization-activated cyclic nucleotide-gated 1 independent grid cell-phase precession in mice.

    PubMed

    Eggink, Hannah; Mertens, Paul; Storm, Eline; Giocomo, Lisa M

    2014-03-01

    Cell assemblies code information in both the temporal and spatial domain. One tractable example of temporal coding is the phenomenon of phase precession. In medial entorhinal cortex, theta-phase precession is observed in spatially specific grid cells, with grid spike-times shifting to earlier phases of the extracellular theta rhythm as the animal passes through the grid field. Although the exact mechanisms underlying spatial-temporal coding remain unknown, computational work points to single-cell oscillatory activity as a biophysical mechanism capable of producing phase precession. Support for this idea comes from observed correlations between single-cell resonance and entorhinal neurons characterized by phase precession. Here, we take advantage of the absence of single-cell theta-frequency resonance in hyperpolarization-activated cyclic nucleotide-gated (HCN) 1 knockout (KO) mice to examine the relationship between intrinsic rhythmicity and phase precession. We find phase precession is highly comparable between forebrain-restricted HCN1 KO and wild-type mice. Grid fields in HCN1 KO mice display more experience-dependent asymmetry however, consistent with reports of enhanced long-term potentiation in the absence of HCN1 and raising the possibility that the loss of HCN1 improves temporal coding via the rate-phase transformation. Combined, our results clarify the role of HCN1 channels in temporal coding and constrain the number of possible mechanisms generating phase precession. © 2013 Wiley Periodicals, Inc.

  17. Effects of pulmonary inhalation on hyperpolarized krypton-83 magnetic resonance T1 relaxation

    NASA Astrophysics Data System (ADS)

    Stupic, K. F.; Elkins, N. D.; Pavlovskaya, G. E.; Repine, J. E.; Meersmann, T.

    2011-07-01

    The 83Kr magnetic resonance (MR) relaxation time T1 of krypton gas in contact with model surfaces was previously found to be highly sensitive to surface composition, surface-to-volume ratio, and surface temperature. The work presented here explored aspects of pulmonary 83Kr T1 relaxation measurements in excised lungs from healthy rats using hyperpolarized (hp) 83Kr with approximately 4.4% spin polarization. MR spectroscopy without spatial resolution was applied to the ex vivo lungs that actively inhale hp 83Kr through a custom designed ventilation system. Various inhalation schemes were devised to study the influence of anatomical dead space upon the measured 83Kr T1 relaxation times. The longitudinal 83Kr relaxation times in the distal airways and the respiratory zones were independent of the lung inhalation volume, with T1 = 1.3 s and T1 = 1.0 s, depending only on the applied inhalation scheme. The obtained data were highly reproducible between different specimens. Further, the 83Kr T1 relaxation times in excised lungs were unaffected by the presence of up to 40% oxygen in the hp gas mixture. The results support the possible importance of 83Kr as a biomarker for evaluating lung function.

  18. HP-Xe to go: Storage and transportation of hyperpolarized 129Xenon

    NASA Astrophysics Data System (ADS)

    Repetto, M.; Zimmer, S.; Allmendinger, F.; Blümler, P.; Doll, M.; Grasdijk, J. O.; Heil, W.; Jungmann, K.; Karpuk, S.; Krause, H.-J.; Offenhäusser, A.; Schmidt, U.; Sobolev, Y.; Willmann, L.

    2016-04-01

    Recently the spin-lattice relaxation time T1 of hyperpolarized (HP)-129Xe was significantly improved by using uncoated and Rb-free storage vessels of GE180 glass. For these cells, a simple procedure was established to obtain reproducible wall relaxation times of about 18 h. Then the limiting relaxation mechanism in pure Xe is due to the coupling between the nuclear spins and the angular momentum of the Xe-Xe van-der-Waals-molecules. This mechanism can be significantly reduced by using different buffer gases of which CO2 was discovered to be the most efficient so far. From these values, it was estimated that for a 1:1 mixture of HP-Xe with CO2 a longitudinal relaxation time of about 7 h can be expected, sufficient to transport HP-Xe from a production to a remote application site. This prediction was verified for such a mixture at a total pressure of about 1 bar in a 10 cm glass cell showing a storage time of T1 ≈ 9 h (for T1wall = (34 ± 9) h) which was transported inside a magnetic box over a distance of about 200 km by car.

  19. Cryogenics free production of hyperpolarized 129Xe and 83Kr for biomedical MRI applications.

    PubMed

    Hughes-Riley, Theodore; Six, Joseph S; Lilburn, David M L; Stupic, Karl F; Dorkes, Alan C; Shaw, Dominick E; Pavlovskaya, Galina E; Meersmann, Thomas

    2013-12-01

    As an alternative to cryogenic gas handling, hyperpolarized (hp) gas mixtures were extracted directly from the spin exchange optical pumping (SEOP) process through expansion followed by compression to ambient pressure for biomedical MRI applications. The omission of cryogenic gas separation generally requires the usage of high xenon or krypton concentrations at low SEOP gas pressures to generate hp (129)Xe or hp (83)Kr with sufficient MR signal intensity for imaging applications. Two different extraction schemes for the hp gasses were explored with focus on the preservation of the nuclear spin polarization. It was found that an extraction scheme based on an inflatable, pressure controlled balloon is sufficient for hp (129)Xe handling, while (83)Kr can efficiently be extracted through a single cycle piston pump. The extraction methods were tested for ex vivo MRI applications with excised rat lungs. Precise mixing of the hp gases with oxygen, which may be of interest for potential in vivo applications, was accomplished during the extraction process using a piston pump. The (83)Kr bulk gas phase T1 relaxation in the mixtures containing more than approximately 1% O2 was found to be slower than that of (129)Xe in corresponding mixtures. The experimental setup also facilitated (129)Xe T1 relaxation measurements as a function of O2 concentration within excised lungs.

  20. Cryogenics free production of hyperpolarized 129Xe and 83Kr for biomedical MRI applications

    NASA Astrophysics Data System (ADS)

    Hughes-Riley, Theodore; Six, Joseph S.; Lilburn, David M. L.; Stupic, Karl F.; Dorkes, Alan C.; Shaw, Dominick E.; Pavlovskaya, Galina E.; Meersmann, Thomas

    2013-12-01

    As an alternative to cryogenic gas handling, hyperpolarized (hp) gas mixtures were extracted directly from the spin exchange optical pumping (SEOP) process through expansion followed by compression to ambient pressure for biomedical MRI applications. The omission of cryogenic gas separation generally requires the usage of high xenon or krypton concentrations at low SEOP gas pressures to generate hp 129Xe or hp 83Kr with sufficient MR signal intensity for imaging applications. Two different extraction schemes for the hp gasses were explored with focus on the preservation of the nuclear spin polarization. It was found that an extraction scheme based on an inflatable, pressure controlled balloon is sufficient for hp 129Xe handling, while 83Kr can efficiently be extracted through a single cycle piston pump. The extraction methods were tested for ex vivo MRI applications with excised rat lungs. Precise mixing of the hp gases with oxygen, which may be of interest for potential in vivo applications, was accomplished during the extraction process using a piston pump. The 83Kr bulk gas phase T1 relaxation in the mixtures containing more than approximately 1% O2 was found to be slower than that of 129Xe in corresponding mixtures. The experimental setup also facilitated 129Xe T1 relaxation measurements as a function of O2 concentration within excised lungs.

  1. Effects of Diffusion Time on Short-Range Hyperpolarized 3He Diffusivity Measurements in Emphysema

    SciTech Connect

    Gierada, David S.; Woods, Jason C.; Bierhals, Andrew J.; Bartel, Seth T.; Ritter, Jon H.; Choong, Cliff K.; Das, Nitin A.; Hong, Cheng; Pilgram, Thomas K.; Chang, Yulin V.; Jacob, Rick E.; Hogg, James C.; Battafarano, Richard J.; Cooper, Joel D.; Meyers, Bryan F.; Patterson, G Alexander; Yablonskiy, Dmitriy A.; Conradi, Mark S.

    2009-09-28

    Purpose: To characterize the effect of diffusion time on short-range hyperpolarized 3He MR diffusion measurements across a wide range of emphysema severity. Materials and Methods: 3He diffusion MR imaging was performed on 19 lungs or lobes resected from 18 subjects with varying degrees of emphysema using 3 diffusion times (1.6 msec, 5 msec, and 10 msec) at constant b value. Emphysema severity was quantified as the mean apparent diffusion coefficient (ADC) and as the percentage of pixels with ADC higher than multiple thresholds from 0.30-0.55 cm2/sec (ADC index). Quantitative histology (mean linear intercept) was obtained in 10 of the lung specimens from 10 of the subjects. Results: The mean ADCs with diffusion times of 1.6, 5.0, and 10.0 msec were 0.46, 0.40, and 0.37 cm2/sec, respectively (P <0.0001, ANOVA). There was no relationship between the ADC magnitude and the effect of diffusion time on ADC values. Mean linear intercept correlated with ADC (r=0.91-0.94, P<0.001) and ADC index (r=0.78-0.92, P<0.01) at all diffusion times.

  2. Universal Long-Time Behavior of Nuclear Spin Decays in Solid Hyperpolarized Xenon

    NASA Astrophysics Data System (ADS)

    Saam, Brian; Morgan, Steven W.; Fine, Boris V.

    2009-05-01

    We have observed a universal long-time behavior of ^129Xe FIDs and solid echoes in polycrystalline hyperpolarized xenon at 77 K. In all cases, a decay of the form F(t) = Ae^-γt(φt + φ) sets in after just a few times T2; the behavior is universal in the sense that the decay constant γ and the beat frequency φ, which together characterize the long-time decay are the same for the FID and for solid echoes having different interpulse delay times τ. These findings reveal a fundamental property of nuclear spin dynamics and are thus relevant to theoretical efforts that have been ongoing for decades to understand NMR lineshapes in solids. Moreover, the functional form and universality of this behavior were previously predicted on the basis of analogy with resonances in classical chaotic systems [2]. While we expect this behavior to be characteristic of nuclear-spin solids in general, ^129Xe is an ideal system to examine it with high precision because of the relatively long T2 1 ms and because spin-exchange optical pumping can be used to achieve greatly enhanced magnetization, allowing precise examination of the decay over 3-4 orders of magnitude. [1] S.W. Morgan, et al., PRL 101, 067601 (2008). [2] B.V. Fine, PRL 94, 247601 (2005).

  3. Membrane potential hyperpolarization in Mammalian cardiac cells by synchronization modulation of Na/K pumps.

    PubMed

    Chen, Wei; Dando, Robin

    2008-02-01

    In previously reported work, we developed a new technique, synchronization modulation, to electrically activate Na/K pump molecules. The fundamental mechanism involved in this technique is a dynamic entrainment procedure of the pump molecules, carried out in a stepwise pattern. The entrainment procedure consists of two steps: synchronization and modulation. We theoretically predicted that the pump functions can be activated exponentially as a function of the membrane potential. We have experimentally demonstrated synchronization of the Na/K pump molecules and acceleration of their pumping rates by many fold through use of voltage-clamp techniques, directly monitoring the pump currents. We further applied this technique to intact skeletal muscle fibers from amphibians and found significant effects on the membrane resting potential. Here, we extend our study to intact mammalian cardiomyocytes. We employed a noninvasive confocal microscopic fluorescent imaging technique to monitor electric field-induced changes in ionic concentration gradient and membrane resting potential. Our results further confirm that the well-designed synchronization modulation electric field can effectively accelerate the Na/K pumping rate, increasing the ionic concentration gradient across the cell membrane and hyperpolarizing the membrane resting potential.

  4. Effects of pulmonary inhalation on hyperpolarized krypton-83 magnetic resonance T1 relaxation.

    PubMed

    Stupic, K F; Elkins, N D; Pavlovskaya, G E; Repine, J E; Meersmann, T

    2011-07-07

    The (83)Kr magnetic resonance (MR) relaxation time T(1) of krypton gas in contact with model surfaces was previously found to be highly sensitive to surface composition, surface-to-volume ratio, and surface temperature. The work presented here explored aspects of pulmonary (83)Kr T(1) relaxation measurements in excised lungs from healthy rats using hyperpolarized (hp) (83)Kr with approximately 4.4% spin polarization. MR spectroscopy without spatial resolution was applied to the ex vivo lungs that actively inhale hp (83)Kr through a custom designed ventilation system. Various inhalation schemes were devised to study the influence of anatomical dead space upon the measured (83)Kr T(1) relaxation times. The longitudinal (83)Kr relaxation times in the distal airways and the respiratory zones were independent of the lung inhalation volume, with T(1) = 1.3 s and T(1) = 1.0 s, depending only on the applied inhalation scheme. The obtained data were highly reproducible between different specimens. Further, the (83)Kr T(1) relaxation times in excised lungs were unaffected by the presence of up to 40% oxygen in the hp gas mixture. The results support the possible importance of (83)Kr as a biomarker for evaluating lung function.

  5. Hyperpolarized xenon-based molecular sensors for label-free detection of analytes.

    PubMed

    Garimella, Praveena D; Meldrum, Tyler; Witus, Leah S; Smith, Monica; Bajaj, Vikram S; Wemmer, David E; Francis, Matthew B; Pines, Alexander

    2014-01-08

    Nuclear magnetic resonance (NMR) can reveal the chemical constituents of a complex mixture without resorting to chemical modification, separation, or other perturbation. Recently, we and others have developed magnetic resonance agents that report on the presence of dilute analytes by proportionately altering the response of a more abundant or easily detected species, a form of amplification. One example of such a sensing medium is xenon gas, which is chemically inert and can be optically hyperpolarized, a process that enhances its NMR signal by up to 5 orders of magnitude. Here, we use a combinatorial synthetic approach to produce xenon magnetic resonance sensors that respond to small molecule analytes. The sensor responds to the ligand by producing a small chemical shift change in the Xe NMR spectrum. We demonstrate this technique for the dye, Rhodamine 6G, for which we have an independent optical assay to verify binding. We thus demonstrate that specific binding of a small molecule can produce a xenon chemical shift change, suggesting a general approach to the production of xenon sensors targeted to small molecule analytes for in vitro assays or molecular imaging in vivo.

  6. Electrophysiological characterization of Grueneberg ganglion olfactory neurons: spontaneous firing, sodium conductance, and hyperpolarization-activated currents.

    PubMed

    Liu, Cambrian Y; Xiao, Cheng; Fraser, Scott E; Lester, Henry A; Koos, David S

    2012-09-01

    Mammals rely on their acute olfactory sense for their survival. The most anterior olfactory subsystem in the nose, the Grueneberg ganglion (GG), plays a role in detecting alarm pheromone, cold, and urinary compounds. GG neurons respond homogeneously to these stimuli with increases in intracellular [Ca(2+)] or transcription of immediate-early genes. In this electrophysiological study, we used patch-clamp techniques to characterize the membrane properties of GG neurons. Our results offer evidence of functional heterogeneity in the GG. GG neurons fire spontaneously and independently in several stable patterns, including phasic and repetitive single-spike modes of discharge. Whole cell recordings demonstrated two distinct voltage-gated fast-inactivating Na(+) currents with different steady-state voltage dependencies and different sensitivities to tetrodotoxin. Hodgkin-Huxley simulations showed that these Na(+) currents confer dual mechanisms of action potential generation and contribute to different firing patterns. Additionally, GG neurons exhibited hyperpolarization-activated inward currents that modulated spontaneous firing in vitro. Thus, in GG neurons, the heterogeneity of firing patterns is linked to the unusual repertoire of ionic currents. The membrane properties described here will aid the interpretation of chemosensory function in the GG.

  7. Hyperpolarization-activated current (I(h)) in ganglion-cell photoreceptors.

    PubMed

    Van Hook, Matthew J; Berson, David M

    2010-12-20

    Intrinsically photosensitive retinal ganglion cells (ipRGCs) express the photopigment melanopsin and serve as the primary retinal drivers of non-image-forming visual functions such as circadian photoentrainment, the pupillary light reflex, and suppression of melatonin production in the pineal. Past electrophysiological studies of these cells have focused on their intrinsic photosensitivity and synaptic inputs. Much less is known about their voltage-gated channels and how these might shape their output to non-image-forming visual centers. Here, we show that rat ipRGCs retrolabeled from the suprachiasmatic nucleus (SCN) express a hyperpolarization-activated inwardly-rectifying current (I(h)). This current is blocked by the known I(h) blockers ZD7288 and extracellular cesium. As in other systems, including other retinal ganglion cells, I(h) in ipRGCs is characterized by slow kinetics and a slightly greater permeability for K(+) than for Na(+). Unlike in other systems, however, I(h) in ipRGCs apparently does not actively contribute to resting membrane potential. We also explore non-specific effects of the common I(h) blocker ZD7288 on rebound depolarization and evoked spiking and discuss possible functional roles of I(h) in non-image-forming vision. This study is the first to characterize I(h) in a well-defined population of retinal ganglion cells, namely SCN-projecting ipRGCs.

  8. Hyperpolarization-Activated Current Induces Period-Doubling Cascades and Chaos in a Cold Thermoreceptor Model

    PubMed Central

    Xu, Kesheng; Maidana, Jean P.; Caviedes, Mauricio; Quero, Daniel; Aguirre, Pablo; Orio, Patricio

    2017-01-01

    In this article, we describe and analyze the chaotic behavior of a conductance-based neuronal bursting model. This is a model with a reduced number of variables, yet it retains biophysical plausibility. Inspired by the activity of cold thermoreceptors, the model contains a persistent Sodium current, a Calcium-activated Potassium current and a hyperpolarization-activated current (Ih) that drive a slow subthreshold oscillation. Driven by this oscillation, a fast subsystem (fast Sodium and Potassium currents) fires action potentials in a periodic fashion. Depending on the parameters, this model can generate a variety of firing patterns that includes bursting, regular tonic and polymodal firing. Here we show that the transitions between different firing patterns are often accompanied by a range of chaotic firing, as suggested by an irregular, non-periodic firing pattern. To confirm this, we measure the maximum Lyapunov exponent of the voltage trajectories, and the Lyapunov exponent and Lempel-Ziv's complexity of the ISI time series. The four-variable slow system (without spiking) also generates chaotic behavior, and bifurcation analysis shows that this is often originated by period doubling cascades. Either with or without spikes, chaos is no longer generated when the Ih is removed from the system. As the model is biologically plausible with biophysically meaningful parameters, we propose it as a useful tool to understand chaotic dynamics in neurons. PMID:28344550

  9. Hyperpolarization induces a long-term increase in the spontaneous firing rate of cerebellar Golgi cells

    PubMed Central

    Hull, Court; Chu, YunXiang; Thanawala, Monica; Regehr, Wade G.

    2013-01-01

    Golgi cells (GoCs) are inhibitory interneurons that influence the cerebellar cortical response to sensory input by regulating the excitability of the granule cell layer. While GoC inhibition is essential for normal motor coordination, little is known about the circuit dynamics that govern the activity of these cells. In particular, while GoC spontaneous spiking influences the extent of inhibition and gain throughout the granule cell layer, it is not known whether this spontaneous activity can be modulated in a long-term manner. Here we describe a form of long-term plasticity that regulates the spontaneous firing rate of GoCs in the rat cerebellar cortex. We find that membrane hyperpolarization, either by mGluR2 activation of potassium channels, or by somatic current injection, induces a long-lasting increase in GoC spontaneous firing. This spike rate plasticity appears to result from a strong reduction in the spike afterhyperpolarization (AHP). Pharmacological manipulations suggest the involvement of calcium-calmodulin dependent kinase II (CaMKII) and calcium-activated potassium channels in mediating these firing rate increases. As a consequence of this plasticity, GoC spontaneous spiking is selectively enhanced, but the gain of evoked spiking is unaffected. Hence this plasticity is well-suited for selectively regulating the tonic output of GoCs rather than their sensory-evoked responses. PMID:23554471

  10. Continuous Problem of Function Continuity

    ERIC Educational Resources Information Center

    Jayakody, Gaya; Zazkis, Rina

    2015-01-01

    We examine different definitions presented in textbooks and other mathematical sources for "continuity of a function at a point" and "continuous function" in the context of introductory level Calculus. We then identify problematic issues related to definitions of continuity and discontinuity: inconsistency and absence of…

  11. Action potential repolarization and a fast after-hyperpolarization in rat hippocampal pyramidal cells.

    PubMed

    Storm, J F

    1987-04-01

    1. The repolarization of the action potential, and a fast after-hyperpolarization (a.h.p.) were studied in CA1 pyramidal cells (n = 76) in rat hippocampal slices (28-37 degrees C). Single spikes were elicited by brief (1-3 ms) current pulses, at membrane potentials close to rest (-60 to -70 mV). 2. Each action potential was followed by four after-potentials: (a) the fast a.h.p., lasting 2-5 ms; (b) an after-depolarization; (c) a medium a.h.p., (50-100 ms); and (d) a slow a.h.p. (1-2 s). Both the fast a.h.p. and the slow a.h.p. (but not the medium a.h.p.) were inhibited by Ca2+-free medium or Ca2+-channel blockers (Co2+, Mn2+ or Cd2+); but tetraethylammonium (TEA; 0.5-2 nM) blocked only the fast a.h.p., and noradrenaline (2-5 microM) only the slow a.h.p. This suggests that two Ca2+-activated K+ currents were involved: a fast, TEA-sensitive one (IC) underlying the fast a.h.p., and a slow noradrenaline-sensitive one (IAHP) underlying the slow a.h.p. 3. Like the fast a.h.p., spike repolarization seems to depend on a Ca2+-dependent K+ current of the fast, TEA-sensitive kind (IC). The repolarization was slowed by Ca2+-free medium, Co2+, Mn2+, Cd2+, or TEA, but not by noradrenaline. Charybdotoxin (CTX; 30 nM), a scorpion toxin which blocks the large-conductance Ca2+-activated K+ channel in muscle, had a similar effect to TEA. The effects of TEA and Cd2+ (or Mn2+) showed mutual occlusion. Raising the external K+ concentration reduced the fast a.h.p. and slowed the spike repolarization, whereas Cl- loading of the cell was ineffective. 4. The transient K+ current, IA, seems also to contribute to spike repolarization, because: (a) 4-aminopyridine (4-AP; 0.1 mM), which blocks IA, slowed the spike repolarization; (b) depolarizing pre-pulses, which inactivate IA, had a similar effect; (c) hyperpolarizing pre-pulses speeded up the spike repolarization; (d) the effects of 4-AP and pre-pulses persisted during Ca2+ blockade (like IA); and (e) depolarizing pre-pulses reduced the

  12. Optimized production of hyperpolarized 129Xe at 2 bars for in vivo lung magnetic resonance imaging

    NASA Astrophysics Data System (ADS)

    Norquay, Graham; Parnell, Steven R.; Xu, Xiaojun; Parra-Robles, Juan; Wild, Jim M.

    2013-01-01

    In this work, the production rate of a spin-exchange optical pumping 129Xe gas polarizer was optimized for routine generation of hyperpolarized 129Xe for in vivo lung MRI. This system uses a narrow (˜ 0.1 nm linewidth), tuneable external cavity laser (operating at ˜25 W) for SEOP of 3% gas mixtures of Xe inside a mid-pressure (2 bars) cell of 491 cm3 volume. Under this regime, theoretical and experimentally measured 129Xe polarizations were calculated to be 24% and 12%, respectively, for a gas flow rate of 300 sccm and a cell temperature of 373 K. The photon efficiency was evaluated, yielding theoretical and experimental values of 0.039 and 0.046, respectively. The theoretical efficiency was calculated from spin-exchange and spin-destruction cross sections and the experimental photon efficiency was measured under flow for a gas-cell residency time equal to an empirically determined spin-exchange time of 45 s. In addition, details of the Xe freeze-out process were analyzed with a model of polarization decay during Xe accumulation in the frozen phase, where a T1 of 87 ± 2 min was observed. To demonstrate the system's application, in vivo lung magnetic resonance images (signal-to-noise ratio ˜ 50 from a voxel of 15 mm× 4 mm× 4 mm) were acquired using modest volumes (<400 ml) of isotopically enriched (86% 129Xe) Xe gas polarized to >10%. Despite the experimental polarization being a factor of 2 lower than the predicted polarization for typical operating parameters, the system is close to the theoretical photon efficiency and the system has so far produced polarized gas for more than 100 in vivo 129Xe lung imaging studies.

  13. Strategies for the hyperpolarization of acetonitrile and related ligands by SABRE.

    PubMed

    Mewis, Ryan E; Green, Richard A; Cockett, Martin C R; Cowley, Michael J; Duckett, Simon B; Green, Gary G R; John, Richard O; Rayner, Peter J; Williamson, David C

    2015-01-29

    We report on a strategy for using SABRE (signal amplification by reversible exchange) for polarizing (1)H and (13)C nuclei of weakly interacting ligands which possess biologically relevant and nonaromatic motifs. We first demonstrate this via the polarization of acetonitrile, using Ir(IMes)(COD)Cl as the catalyst precursor, and confirm that the route to hyperpolarization transfer is via the J-coupling network. We extend this work to the polarization of propionitrile, benzylnitrile, benzonitrile, and trans-3-hexenedinitrile in order to assess its generality. In the (1)H NMR spectrum, the signal for acetonitrile is enhanced 8-fold over its thermal counterpart when [Ir(H)2(IMes)(MeCN)3](+) is the catalyst. Upon addition of pyridine or pyridine-d5, the active catalyst changes to [Ir(H)2(IMes)(py)2(MeCN)](+) and the resulting acetonitrile (1)H signal enhancement increases to 20- and 60-fold, respectively. In (13)C NMR studies, polarization transfers optimally to the quaternary (13)C nucleus of MeCN while the methyl (13)C is hardly polarized. Transfer to (13)C is shown to occur first via the (1)H-(1)H coupling between the hydrides and the methyl protons and then via either the (2)J or (1)J couplings to the respective (13)Cs, of which the (2)J route is more efficient. These experimental results are rationalized through a theoretical treatment which shows excellent agreement with experiment. In the case of MeCN, longitudinal two-spin orders between pairs of (1)H nuclei in the three-spin methyl group are created. Two-spin order states, between the (1)H and (13)C nuclei, are also created, and their existence is confirmed for Me(13)CN in both the (1)H and (13)C NMR spectra using the Only Parahydrogen Spectroscopy protocol.

  14. In vivo lung morphometry with hyperpolarized 3He diffusion MRI: theoretical background.

    PubMed

    Sukstanskii, A L; Yablonskiy, D A

    2008-02-01

    MRI-based study of (3)He gas diffusion in lungs may provide important information on lung microstructure. Lung acinar airways can be described in terms of cylinders covered with alveolar sleeve [Haefeli-Bleuer, Weibel, Anat. Rec. 220 (1988) 401]. For relatively short diffusion times (on the order of a few ms) this geometry allows description of the (3)He diffusion attenuated MR signal in lungs in terms of two diffusion coefficients-longitudinal (D(L)) and transverse (D(T)) with respect to the individual acinar airway axis [Yablonskiy et al., PNAS 99 (2002) 3111]. In this paper, empirical relationships between D(L) and D(T) and the geometrical parameters of airways and alveoli are found by means of computer Monte Carlo simulations. The effects of non-Gaussian signal behavior (dependence of D(L) and D(T) on b-value) are also taken into account. The results obtained are quantitatively valid in the physiologically important range of airway parameters characteristic of healthy lungs and lungs with mild emphysema. In lungs with advanced emphysema, the results provide only "apparent" characteristics but still could potentially be used to evaluate emphysema progression. This creates a basis for in vivo lung morphometry-evaluation of the geometrical parameters of acinar airways from hyperpolarized (3)He diffusion MRI, despite the airways being too small to be resolved by direct imaging. These results also predict a rather substantial dependence of (3)He ADC on the experimentally-controllable diffusion time, Delta. If Delta is decreased from 3 ms to 1 ms, the ADC in normal human lungs may increase by almost 50%. This effect should be taken into account when comparing experimental data obtained with different pulse sequences.

  15. Mitochondrial Hyperpolarization and ATP Depletion in Patients With Systemic Lupus Erythematosus

    PubMed Central

    Gergely, Peter; Grossman, Craig; Niland, Brian; Puskas, Ferenc; Neupane, Hom; Allam, Fatme; Banki, Katalin; Phillips, Paul E.; Perl, Andras

    2014-01-01

    Objective Peripheral blood lymphocytes (PBLs) from systemic lupus erythematosus (SLE) patients exhibit increased spontaneous and diminished activation-induced apoptosis. We tested the hypothesis that key biochemical checkpoints, the mitochondrial transmembrane potential (ΔΨm) and production of reactive oxygen intermediates (ROIs), mediate the imbalance of apoptosis in SLE. Methods We assessed the ΔΨm with potentiometric dyes, measured ROI production with oxidation-sensitive fluorochromes, and monitored cell death by annexin V and propidium iodide staining of lymphocytes, using flow cytometry. Intracellular glutathione levels were measured by high-performance liquid chromatography, while ATP and ADP levels were assessed by the luciferin–luciferase assay. Results Both ΔΨm and ROI production were elevated in the 25 SLE patients compared with the 25 healthy subjects and the 10 rheumatoid arthritis patients. Intracellular glutathione contents were diminished, suggesting increased utilization of reducing equivalents in SLE. H2O2, a precursor of ROIs, increased ΔΨm and caused apoptosis in normal PBLs. In contrast, H2O2-induced apoptosis and ΔΨm elevation were diminished, particularly in T cells, and the rate of necrotic cell death was increased in patients with SLE. The intracellular ATP content and the ATP:ADP ratio were reduced and correlated with the ΔΨm elevation in lupus. CD3:CD28 costimulation led to transient elevation of the ΔΨm, followed by ATP depletion, and sensitization of normal PBLs to H2O2-induced necrosis. Depletion of ATP by oligomycin, an inhibitor of F0F1–ATPase, had similar effects. Conclusion T cell activation and apoptosis are mediated by ΔΨm elevation and increased ROI production. Mitochondrial hyperpolarization and the resultant ATP depletion sensitize T cells for necrosis, which may significantly contribute to inflammation in patients with SLE. PMID:11817589

  16. Endothelium-Derived Hyperpolarizing Factor Mediates Bradykinin Stimulated Tissue Plasminogen Activator Release In Humans

    PubMed Central

    Rahman, Ayaz M.; Murrow, Jonathan R.; Ozkor, Muhiddin A.; Kavtaradze, Nino; Lin, Ji; De Staercke, Christine; Hooper, W. Craig; Manatunga, Amita; Hayek, Salim; Quyyumi, Arshed A.

    2014-01-01

    Aims Bradykinin stimulates tissue plasminogen activator (t-PA) release from human endothelium. Although bradykinin stimulates both nitric oxide and endothelium-derived hyperpolarizing factor (EDHF) release, the role of EDHF in t-PA release remains unexplored. This study sought to determine the mechanisms of bradykinin-stimulated t-PA release in the forearm vasculature of healthy human subjects. Methods In 33 healthy subjects (age 40.3±1.9 years) forearm blood flow (FBF) and t-PA release were measured at rest, and after intra-arterial infusions of bradykinin (400ng/min) and sodium nitroprusside (3.2 mg/min). Measurements were repeated after intra-arterial infusion of TEA (1 μmol/min), fluconazole (0.4 μmol.min-1.L-1), and NG-monomethyl-L-arginine (L-NMMA, 8 μmol/min) to block nitric oxide, and their combination in separate studies. Results Bradykinin significantly increased net t-PA release across the forearm (P<0.0001). Fluconazole attenuated both bradykinin-mediated vasodilation (-23.3±2.7% FBF, P<0.0001) and t-PA release (from 50.9±9.0 to 21.3±8.9 ng/min/100ml, P=0.02). TEA attenuated FBF (-14.7±3.2%, P=0.002) and abolished bradykinin-stimulated t-PA release (from 22.9+5.7 to - 0.8±3.6 ng/min/100ml, P=0.0002). L-NMMA attenuated FBF (P<0.0001), but did not inhibit bradykinin-induced t-PA release (P=NS). Conclusion Bradykinin-stimulated t-PA release is partly due to cytochrome P450-derived epoxides, and is inhibited by K+ca channel blockade. Thus, bradykinin stimulates both EDHF-dependent vasodilation and t-PA release. PMID:24925526

  17. Clcn2 encodes the hyperpolarization-activated chloride channel in the ducts of mouse salivary glands

    PubMed Central

    Romanenko, Victor G.; Nakamoto, Tetsuji; Catalán, Marcelo A.; Gonzalez-Begne, Mireya; Schwartz, George J.; Jaramillo, Yasna; Sepúlveda, Francisco V.; Figueroa, Carlos D.; Melvin, James E.

    2008-01-01

    Transepithelial Cl− transport in salivary gland ducts is a major component of the ion reabsorption process, the final stage of saliva production. It was previously demonstrated that a Cl− current with the biophysical properties of ClC-2 channels dominates the Cl− conductance of unstimulated granular duct cells in the mouse submandibular gland. This inward-rectifying Cl− current is activated by hyperpolarization and elevated intracellular Cl− concentration. Here we show that ClC-2 immunolocalized to the basolateral region of acinar and duct cells in mouse salivary glands, whereas its expression was most robust in granular and striated duct cells. Consistent with this observation, nearly 10-fold larger ClC-2-like currents were observed in granular duct cells than the acinar cells obtained from submandibular glands. The loss of inward-rectifying Cl− current in cells from Clcn2−/− mice confirmed the molecular identity of the channel responsible for these currents as ClC-2. Nevertheless, both in vivo and ex vivo fluid secretion assays failed to identify significant changes in the ion composition, osmolality, or salivary flow rate of Clcn2−/− mice. Additionally, neither a compensatory increase in Cftr Cl− channel protein expression nor in Cftr-like Cl− currents were detected in Clcn2 null mice, nor did it appear that ClC-2 was important for blood-organ barrier function. We conclude that ClC-2 is the inward-rectifying Cl− channel in duct cells, but its expression is not apparently required for the ion reabsorption or the barrier function of salivary ductal epithelium. PMID:18801913

  18. Src tyrosine kinase alters gating of hyperpolarization-activated HCN4 pacemaker channel through Tyr531.

    PubMed

    Li, Chen-Hong; Zhang, Qi; Teng, Bunyen; Mustafa, S Jamal; Huang, Jian-Ying; Yu, Han-Gang

    2008-01-01

    We recently discovered that the constitutively active Src tyrosine kinase can enhance hyperpolarization-activated, cyclic nucleotide-gated (HCN) 4 channel activity by binding to the channel protein. To investigate the mechanism of modulation by Src of HCN channels, we studied the effects of a selective inhibitor of Src tyrosine kinase, 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2), on HCN4 and its mutant channels expressed in HEK 293 cells by using a whole cell patch-clamp technique. We found that PP2 can inhibit HCN4 currents by negatively shifting the voltage dependence of channel activation, decreasing the whole cell channel conductance, and slowing activation and deactivation kinetics. Screening putative tyrosine residues subject to phosphorylation yielded two candidates: Tyr(531) and Tyr(554). Substituting HCN4-Tyr(531) with phenylalanine largely abolished the effects of PP2 on HCN4 channels. Replacing HCN4-Tyr(554) with phenylalanine did not abolish the effects of PP2 on voltage-dependent activation but did eliminate PP2-induced slowing of channel kinetics. The inhibitory effects of HCN channels associated with reduced Src tyrosine activity is confirmed in HL-1 cardiomyocytes. Finally, we found that PP2 can decrease the heart rate in a mouse model. These results demonstrate that Src tyrosine kinase enhances HCN4 currents by shifting their activation to more positive potentials and increasing the whole cell channel conductance as well as speeding the channel kinetics. The tyrosine residue that mediates most of Src's actions on HCN4 channels is Tyr(531).

  19. Kinetic modeling of hyperpolarized 13C 1-pyruvate metabolism in normal rats and TRAMP mice

    NASA Astrophysics Data System (ADS)

    Zierhut, Matthew L.; Yen, Yi-Fen; Chen, Albert P.; Bok, Robert; Albers, Mark J.; Zhang, Vickie; Tropp, Jim; Park, Ilwoo; Vigneron, Daniel B.; Kurhanewicz, John; Hurd, Ralph E.; Nelson, Sarah J.

    2010-01-01

    PurposeTo investigate metabolic exchange between 13C 1-pyruvate, 13C 1-lactate, and 13C 1-alanine in pre-clinical model systems using kinetic modeling of dynamic hyperpolarized 13C spectroscopic data and to examine the relationship between fitted parameters and dose-response. Materials and methodsDynamic 13C spectroscopy data were acquired in normal rats, wild type mice, and mice with transgenic prostate tumors (TRAMP) either within a single slice or using a one-dimensional echo-planar spectroscopic imaging (1D-EPSI) encoding technique. Rate constants were estimated by fitting a set of exponential equations to the dynamic data. Variations in fitted parameters were used to determine model robustness in 15 mm slices centered on normal rat kidneys. Parameter values were used to investigate differences in metabolism between and within TRAMP and wild type mice. ResultsThe kinetic model was shown here to be robust when fitting data from a rat given similar doses. In normal rats, Michaelis-Menten kinetics were able to describe the dose-response of the fitted exchange rate constants with a 13.65% and 16.75% scaled fitting error (SFE) for kpyr→lac and kpyr→ala, respectively. In TRAMP mice, kpyr→lac increased an average of 94% after up to 23 days of disease progression, whether the mice were untreated or treated with casodex. Parameters estimated from dynamic 13C 1D-EPSI data were able to differentiate anatomical structures within both wild type and TRAMP mice. ConclusionsThe metabolic parameters estimated using this approach may be useful for in vivo monitoring of tumor progression and treatment efficacy, as well as to distinguish between various tissues based on metabolic activity.

  20. Novel blockers of hyperpolarization-activated current with isoform selectivity in recombinant cells and native tissue.

    PubMed

    Del Lungo, Martina; Melchiorre, Michele; Guandalini, Luca; Sartiani, Laura; Mugelli, Alessandro; Koncz, Istvan; Szel, Tamas; Varro, Andras; Romanelli, Maria Novella; Cerbai, Elisabetta

    2012-05-01

    BACKGROUND AND PURPOSE Selective hyperpolarization activated, cyclic nucleotide-gated channel (HCN) blockers represent an important therapeutic goal due to the wide distribution and multiple functions of these proteins, representing the molecular correlate of f- and h-current (I(f) or I(h) ). Recently, new compounds able to block differentially the homomeric HCN isoforms expressed in HEK293 have been synthesized. In the present work, the electrophysiological and pharmacological properties of these new HCN blockers were characterized and their activities evaluated on native channels. EXPERIMENTAL APPROACH HEK293 cells expressing mHCN1, mHCN2 and hHCN4 isoforms were used to verify channel blockade. Selected compounds were tested on native guinea pig sinoatrial node cells and neurons from mouse dorsal root ganglion (DRG) by patch-clamp recordings and on dog Purkinje fibres by intracellular recordings. KEY RESULTS In HEK293 cells, EC18 was found to be significantly selective for HCN4 and MEL57A for HCN1 at physiological membrane potential. When tested on guinea pig sinoatrial node cells, EC18 (10 µM) maintained its activity, reducing I(f) by 67% at -120 mV, while MEL57A (3 µM) reduced I(f) by 18%. In contrast, in mouse DRG neurons, only MEL57A (30 and 100 µM) significantly reduced I(h) by 60% at -80 mV. In dog cardiac Purkinje fibres, EC18, but not MEL57A, reduced the amplitude and slowed the slope of the spontaneous diastolic depolarization. CONCLUSIONS Our results have identified novel and highly selective HCN isoform blockers, EC18 and MEL57A; the selectivity found in recombinant system was maintained in various tissues expressing different HCN isoforms.

  1. Hyperpolarized Gas Diffusion MRI for the Study of Atelectasis and Acute Respiratory Distress Syndrome

    PubMed Central

    Cereda, Maurizio; Xin, Yi; Kadlecek, Stephen; Hamedani, Hooman; Rajaei, Jennia; Clapp, Justin; Rizi, Rahim R.

    2014-01-01

    Considerable uncertainty remains about the best ventilator strategies for the mitigation of atelectasis and associated airspace stretch in patients with acute respiratory distress syndrome (ARDS). In addition to several immediate physiological effects, atelectasis increases the risk of ventilator-associated lung injury (VALI), which has been shown to significantly worsen ARDS outcomes. A number of lung imaging techniques have made substantial headway in clarifying the mechanisms of atelectasis. This paper reviews the contributions of CT, PET, and conventional MRI to understanding this phenomenon. In doing so, it also reveals several important shortcomings inherent to each of these approaches. Once these shortcomings have been made apparent, we describe how hyperpolarized gas magnetic resonance imaging (HP MRI)—a technique that is uniquely able to assess responses to mechanical ventilation and lung injury in peripheral airspaces—is poised to fill several of these knowledge gaps. The HP-MRI-derived apparent diffusion coefficient (ADC) quantifies the restriction of 3He diffusion by peripheral airspaces, thereby obtaining pulmonary structural information at an extremely small scale. Lastly, this paper reports the results of a series of experiments that measured ADC in mechanically ventilated rats in order to investigate (i) the effect of atelectasis on ventilated airspaces; (ii) the relationship between positive end-expiratory pressure (PEEP), hysteresis, and the dimensions of peripheral airspaces; and (iii) the ability of PEEP and surfactant to reduce airspace dimensions after lung injury. An increase in ADC was found to be a marker of atelectasis-induced overdistension. With recruitment, higher airway pressures were shown to reduce stretch rather than worsen it. Moving forward, HP MRI has significant potential to shed further light on the atelectatic processes that occur during mechanical ventilation. PMID:24920074

  2. In vivo lung morphometry with hyperpolarized 3He diffusion MRI: Theoretical background

    NASA Astrophysics Data System (ADS)

    Sukstanskii, A. L.; Yablonskiy, D. A.

    2008-02-01

    MRI-based study of 3He gas diffusion in lungs may provide important information on lung microstructure. Lung acinar airways can be described in terms of cylinders covered with alveolar sleeve [Haefeli-Bleuer, Weibel, Anat. Rec. 220 (1988) 401]. For relatively short diffusion times (on the order of a few ms) this geometry allows description of the 3He diffusion attenuated MR signal in lungs in terms of two diffusion coefficients—longitudinal (D) and transverse (D) with respect to the individual acinar airway axis [Yablonskiy et al., PNAS 99 (2002) 3111]. In this paper, empirical relationships between D and D and the geometrical parameters of airways and alveoli are found by means of computer Monte Carlo simulations. The effects of non-Gaussian signal behavior (dependence of D and D on b-value) are also taken into account. The results obtained are quantitatively valid in the physiologically important range of airway parameters characteristic of healthy lungs and lungs with mild emphysema. In lungs with advanced emphysema, the results provide only "apparent" characteristics but still could potentially be used to evaluate emphysema progression. This creates a basis for in vivo lung morphometry—evaluation of the geometrical parameters of acinar airways from hyperpolarized 3He diffusion MRI, despite the airways being too small to be resolved by direct imaging. These results also predict a rather substantial dependence of 3He ADC on the experimentally-controllable diffusion time, Δ. If Δ is decreased from 3 ms to 1 ms, the ADC in normal human lungs may increase by almost 50%. This effect should be taken into account when comparing experimental data obtained with different pulse sequences.

  3. Pathway to Cryogen Free Production of Hyperpolarized Krypton-83 and Xenon-129

    PubMed Central

    Six, Joseph S.; Hughes-Riley, Theodore; Stupic, Karl F.; Pavlovskaya, Galina E.; Meersmann, Thomas

    2012-01-01

    Hyperpolarized (hp) 129Xe and hp 83Kr for magnetic resonance imaging (MRI) are typically obtained through spin-exchange optical pumping (SEOP) in gas mixtures with dilute concentrations of the respective noble gas. The usage of dilute noble gases mixtures requires cryogenic gas separation after SEOP, a step that makes clinical and preclinical applications of hp 129Xe MRI cumbersome. For hp 83Kr MRI, cryogenic concentration is not practical due to depolarization that is caused by quadrupolar relaxation in the condensed phase. In this work, the concept of stopped flow SEOP with concentrated noble gas mixtures at low pressures was explored using a laser with 23.3 W of output power and 0.25 nm linewidth. For 129Xe SEOP without cryogenic separation, the highest obtained MR signal intensity from the hp xenon-nitrogen gas mixture was equivalent to that arising from 15.5±1.9% spin polarized 129Xe in pure xenon gas. The production rate of the hp gas mixture, measured at 298 K, was 1.8 cm3/min. For hp 83Kr, the equivalent of 4.4±0.5% spin polarization in pure krypton at a production rate of 2 cm3/min was produced. The general dependency of spin polarization upon gas pressure obtained in stopped flow SEOP is reported for various noble gas concentrations. Aspects of SEOP specific to the two noble gas isotopes are discussed and compared with current theoretical opinions. A non-linear pressure broadening of the Rb D1 transition was observed and taken into account for the qualitative description of the SEOP process. PMID:23209620

  4. Pathway to cryogen free production of hyperpolarized Krypton-83 and Xenon-129.

    PubMed

    Six, Joseph S; Hughes-Riley, Theodore; Stupic, Karl F; Pavlovskaya, Galina E; Meersmann, Thomas

    2012-01-01

    Hyperpolarized (hp) (129)Xe and hp (83)Kr for magnetic resonance imaging (MRI) are typically obtained through spin-exchange optical pumping (SEOP) in gas mixtures with dilute concentrations of the respective noble gas. The usage of dilute noble gases mixtures requires cryogenic gas separation after SEOP, a step that makes clinical and preclinical applications of hp (129)Xe MRI cumbersome. For hp (83)Kr MRI, cryogenic concentration is not practical due to depolarization that is caused by quadrupolar relaxation in the condensed phase. In this work, the concept of stopped flow SEOP with concentrated noble gas mixtures at low pressures was explored using a laser with 23.3 W of output power and 0.25 nm linewidth. For (129)Xe SEOP without cryogenic separation, the highest obtained MR signal intensity from the hp xenon-nitrogen gas mixture was equivalent to that arising from 15.5±1.9% spin polarized (129)Xe in pure xenon gas. The production rate of the hp gas mixture, measured at 298 K, was 1.8 cm(3)/min. For hp (83)Kr, the equivalent of 4.4±0.5% spin polarization in pure krypton at a production rate of 2 cm(3)/min was produced. The general dependency of spin polarization upon gas pressure obtained in stopped flow SEOP is reported for various noble gas concentrations. Aspects of SEOP specific to the two noble gas isotopes are discussed and compared with current theoretical opinions. A non-linear pressure broadening of the Rb D(1) transition was observed and taken into account for the qualitative description of the SEOP process.

  5. Linking Ventilation Heterogeneity Quantified via Hyperpolarized 3He MRI to Dynamic Lung Mechanics and Airway Hyperresponsiveness.

    PubMed

    Lui, Justin K; Parameswaran, Harikrishnan; Albert, Mitchell S; Lutchen, Kenneth R

    2015-01-01

    Advancements in hyperpolarized helium-3 MRI (HP 3He-MRI) have introduced the ability to render and quantify ventilation patterns throughout the anatomic regions of the lung. The goal of this study was to establish how ventilation heterogeneity relates to the dynamic changes in mechanical lung function and airway hyperresponsiveness in asthmatic subjects. In four healthy and nine mild-to-moderate asthmatic subjects, we measured dynamic lung resistance and lung elastance from 0.1 to 8 Hz via a broadband ventilation waveform technique. We quantified ventilation heterogeneity using a recently developed coefficient of variation method from HP 3He-MRI imaging. Dynamic lung mechanics and imaging were performed at baseline, post-challenge, and after a series of five deep inspirations. AHR was measured via the concentration of agonist that elicits a 20% decrease in the subject's forced expiratory volume in one second compared to baseline (PC20) dose. The ventilation coefficient of variation was correlated to low-frequency lung resistance (R = 0.647, P < 0.0001), the difference between high and low frequency lung resistance (R = 0.668, P < 0.0001), and low-frequency lung elastance (R = 0.547, P = 0.0003). In asthmatic subjects with PC20 values <25 mg/mL, the coefficient of variation at baseline exhibited a strong negative trend (R = -0.798, P = 0.02) to PC20 dose. Our findings were consistent with the notion of peripheral rather than central involvement of ventilation heterogeneity. Also, the degree of AHR appears to be dependent on the degree to which baseline airway constriction creates baseline ventilation heterogeneity. HP 3He-MRI imaging may be a powerful predictor of the degree of AHR and in tracking the efficacy of therapy.

  6. Strategies for the Hyperpolarization of Acetonitrile and Related Ligands by SABRE

    PubMed Central

    2014-01-01

    We report on a strategy for using SABRE (signal amplification by reversible exchange) for polarizing 1H and 13C nuclei of weakly interacting ligands which possess biologically relevant and nonaromatic motifs. We first demonstrate this via the polarization of acetonitrile, using Ir(IMes)(COD)Cl as the catalyst precursor, and confirm that the route to hyperpolarization transfer is via the J-coupling network. We extend this work to the polarization of propionitrile, benzylnitrile, benzonitrile, and trans-3-hexenedinitrile in order to assess its generality. In the 1H NMR spectrum, the signal for acetonitrile is enhanced 8-fold over its thermal counterpart when [Ir(H)2(IMes)(MeCN)3]+ is the catalyst. Upon addition of pyridine or pyridine-d5, the active catalyst changes to [Ir(H)2(IMes)(py)2(MeCN)]+ and the resulting acetonitrile 1H signal enhancement increases to 20- and 60-fold, respectively. In 13C NMR studies, polarization transfers optimally to the quaternary 13C nucleus of MeCN while the methyl 13C is hardly polarized. Transfer to 13C is shown to occur first via the 1H–1H coupling between the hydrides and the methyl protons and then via either the 2J or 1J couplings to the respective 13Cs, of which the 2J route is more efficient. These experimental results are rationalized through a theoretical treatment which shows excellent agreement with experiment. In the case of MeCN, longitudinal two-spin orders between pairs of 1H nuclei in the three-spin methyl group are created. Two-spin order states, between the 1H and 13C nuclei, are also created, and their existence is confirmed for Me13CN in both the 1H and 13C NMR spectra using the Only Parahydrogen Spectroscopy protocol. PMID:25539423

  7. Applying non-uniform grids to evaluating susceptibility from flow-type phenomena: an example of application to Mount Etna

    NASA Astrophysics Data System (ADS)

    Bongolan, Vena Pearl; Lupiano, Valeria; D'Ambrosio, Donato; Rongo, Rocco; Spataro, William; Iovine, Giulio

    2013-04-01

    The hazard induced by dangerous flow-type phenomena - e.g. lava flows, earth flows, debris flows, and debris avalanches - can be assessed by analysing a proper set of simulations of hypothetical events. Non-uniform grids are commonly used to study particular areas of interest in computational domains. Examples of application concern, for instance, the turbulence in a boundary layer. While non-uniform grids frequently appear in adaptive methods, they may also be used in a "static" environment. A purposive sampling method, based on a non-uniform grid of sources coupled with numerical simulations of independent events, has recently been employed to evaluate the hazard induced by flow-type phenomena. An example of application to lava-flows at Mount Etna (Italy) is described in this study. The method aims at refining the spatial distribution of hypothetical eruptive vents with respect to an original uniform grid. The density of eruptive vents has been determined by considering the historical distribution of lateral and eccentric vents, and the distribution of the main faults/structures on the volcano. A higher number of sources marks higher-probability zones of vent opening, based on classes of activation: the number of vents in each class has been set proportionally to the probability of activation of the class. By considering the different types of eruption expected from the considered volcano, based on the historical activity of the past 400 years, a set of simulations per each vent has been performed. The employed model is SCIARA-fv2, a Cellular Automata numerical code recently applied to the same study area for preliminary hazard analyses. In this work, calibration could therefore be skipped, by taking advantage from such experience of tuning of the parameters. Performed simulations have been analysed by a GIS, to verify the number of events affecting each cell of the domain. A probability of occurrence could be assigned to each simulation, based on statistics of

  8. Removal Efficiency of Radioactive Cesium and Iodine Ions by a Flow-Type Apparatus Designed for Electrochemically Reduced Water Production

    PubMed Central

    Hamasaki, Takeki; Nakamichi, Noboru; Teruya, Kiichiro; Shirahata, Sanetaka

    2014-01-01

    The Fukushima Daiichi Nuclear Power Plant accident on March 11, 2011 attracted people’s attention, with anxiety over possible radiation hazards. Immediate and long-term concerns are around protection from external and internal exposure by the liberated radionuclides. In particular, residents living in the affected regions are most concerned about ingesting contaminated foodstuffs, including drinking water. Efficient removal of radionuclides from rainwater and drinking water has been reported using several pot-type filtration devices. A currently used flow-type test apparatus is expected to simultaneously provide radionuclide elimination prior to ingestion and protection from internal exposure by accidental ingestion of radionuclides through the use of a micro-carbon carboxymethyl cartridge unit and an electrochemically reduced water production unit, respectively. However, the removability of radionuclides from contaminated tap water has not been tested to date. Thus, the current research was undertaken to assess the capability of the apparatus to remove radionuclides from artificially contaminated tap water. The results presented here demonstrate that the apparatus can reduce radioactivity levels to below the detection limit in applied tap water containing either 300 Bq/kg of 137Cs or 150 Bq/kg of 125I. The apparatus had a removal efficiency of over 90% for all concentration ranges of radio–cesium and –iodine tested. The results showing efficient radionuclide removability, together with previous studies on molecular hydrogen and platinum nanoparticles as reactive oxygen species scavengers, strongly suggest that the test apparatus has the potential to offer maximum safety against radionuclide-contaminated foodstuffs, including drinking water. PMID:25029447

  9. Removal efficiency of radioactive cesium and iodine ions by a flow-type apparatus designed for electrochemically reduced water production.

    PubMed

    Hamasaki, Takeki; Nakamichi, Noboru; Teruya, Kiichiro; Shirahata, Sanetaka

    2014-01-01

    The Fukushima Daiichi Nuclear Power Plant accident on March 11, 2011 attracted people's attention, with anxiety over possible radiation hazards. Immediate and long-term concerns are around protection from external and internal exposure by the liberated radionuclides. In particular, residents living in the affected regions are most concerned about ingesting contaminated foodstuffs, including drinking water. Efficient removal of radionuclides from rainwater and drinking water has been reported using several pot-type filtration devices. A currently used flow-type test apparatus is expected to simultaneously provide radionuclide elimination prior to ingestion and protection from internal exposure by accidental ingestion of radionuclides through the use of a micro-carbon carboxymethyl cartridge unit and an electrochemically reduced water production unit, respectively. However, the removability of radionuclides from contaminated tap water has not been tested to date. Thus, the current research was undertaken to assess the capability of the apparatus to remove radionuclides from artificially contaminated tap water. The results presented here demonstrate that the apparatus can reduce radioactivity levels to below the detection limit in applied tap water containing either 300 Bq/kg of 137Cs or 150 Bq/kg of 125I. The apparatus had a removal efficiency of over 90% for all concentration ranges of radio-cesium and -iodine tested. The results showing efficient radionuclide removability, together with previous studies on molecular hydrogen and platinum nanoparticles as reactive oxygen species scavengers, strongly suggest that the test apparatus has the potential to offer maximum safety against radionuclide-contaminated foodstuffs, including drinking water.

  10. Continuation Education.

    ERIC Educational Resources Information Center

    Fremont Unified School District, CA.

    The Fremont, California Unified School District organized a continuing education program through a workshop held in the summer of 1968. This paper presents the results of that workshop. Following a statement of philosophy, an outline of the characteristics of the continuation student, and an outline of the functions of the program, an overview of…

  11. Acute porcine renal metabolic effect of endogastric soft drink administration assessed with hyperpolarized [1‐13c]pyruvate

    PubMed Central

    Hansen, Esben Søvsø Szocska; Kjærgaard, Uffe; Bertelsen, Lotte Bonde; Ringgaard, Steffen; Stødkilde‐Jørgensen, Hans

    2015-01-01

    Purpose Our aim was to determine the quantitative reproducibility of metabolic breakdown products in the kidney following intravenous injection of hyperpolarized [1‐13C]pyruvate and secondly to investigate the metabolic effect on the pyruvate metabolism of oral sucrose load using dissolution dynamic nuclear polarization. By this technique, metabolic alterations in several different metabolic related diseases and their metabolic treatment responses can be accessed. Methods In four healthy pigs the lactate‐to‐pyruvate, alanine‐to‐pyruvate and bicarbonate‐to‐pyruvate ratio was measured following administration of regular cola and consecutive injections of hyperpolarized [1‐13C]pyruvate four times within an hour. Results The overall lactate‐to‐pyruvate metabolic profile changed significantly over one hour following an acute sucrose load leading to a significant rise in blood glucose. Conclusion The reproducibility of hyperpolarized magnetic resonance spectroscopy in the healthy pig kidney demonstrated a repeatability of more than 94% for all metabolites and, furthermore, that the pyruvate to lactate conversion and the blood glucose level is elevated following endogastric sucrose administration. Magn Reson Med 74:558–563, 2015. © 2015 The Authors Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial‐NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. PMID:26014387

  12. Modeling non-linear kinetics of hyperpolarized [1-(13)C] pyruvate in the crystalloid-perfused rat heart.

    PubMed

    Mariotti, E; Orton, M R; Eerbeek, O; Ashruf, J F; Zuurbier, C J; Southworth, R; Eykyn, T R

    2016-04-01

    Hyperpolarized (13)C MR measurements have the potential to display non-linear kinetics. We have developed an approach to describe possible non-first-order kinetics of hyperpolarized [1-(13)C] pyruvate employing a system of differential equations that agrees with the principle of conservation of mass of the hyperpolarized signal. Simultaneous fitting to a second-order model for conversion of [1-(13)C] pyruvate to bicarbonate, lactate and alanine was well described in the isolated rat heart perfused with Krebs buffer containing glucose as sole energy substrate, or glucose supplemented with pyruvate. Second-order modeling yielded significantly improved fits of pyruvate-bicarbonate kinetics compared with the more traditionally used first-order model and suggested time-dependent decreases in pyruvate-bicarbonate flux. Second-order modeling gave time-dependent changes in forward and reverse reaction kinetics of pyruvate-lactate exchange and pyruvate-alanine exchange in both groups of hearts during the infusion of pyruvate; however, the fits were not significantly improved with respect to a traditional first-order model. The mechanism giving rise to second-order pyruvate dehydrogenase (PDH) kinetics was explored experimentally using surface fluorescence measurements of nicotinamide adenine dinucleotide reduced form (NADH) performed under the same conditions, demonstrating a significant increase of NADH during pyruvate infusion. This suggests a simultaneous depletion of available mitochondrial NAD(+) (the cofactor for PDH), consistent with the non-linear nature of the kinetics. NADH levels returned to baseline following cessation of the pyruvate infusion, suggesting this to be a transient effect.

  13. ¹H NMR and hyperpolarized ¹³C NMR assays of pyruvate-lactate: a comparative study.

    PubMed

    Hill, Deborah K; Jamin, Yann; Orton, Matthew R; Tardif, Nicolas; Parkes, Harold G; Robinson, Simon P; Leach, Martin O; Chung, Yuen-Li; Eykyn, Thomas R

    2013-10-01

    Pyruvate-lactate exchange is mediated by the enzyme lactate dehydrogenase (LDH) and is central to the altered energy metabolism in cancer cells. The measurement of exchange kinetics using hyperpolarized (13) C NMR has provided a biomarker of response to novel therapeutics. However, the observable signal is restricted to the exchanging hyperpolarized (13) C pools and the endogenous pools of (12) C-labelled metabolites are invisible in these measurements. In this study, we investigated an alternative in vitro (1) H NMR assay, using [3-(13) C]pyruvate, and compared the measured kinetics with a hyperpolarized (13) C NMR assay, using [1-(13) C]pyruvate, under the same conditions in human colorectal carcinoma SW1222 cells. The apparent forward reaction rate constants (kPL ) derived from the two assays showed no significant difference, and both assays had similar reproducibility (kPL  = 0.506 ± 0.054 and kPL  = 0.441 ± 0.090 nmol/s/10(6) cells; mean ± standard deviation; n = 3); (1) H, (13) C assays, respectively). The apparent backward reaction rate constant (kLP ) could only be measured with good reproducibility using the (1) H NMR assay (kLP  = 0.376 ± 0.091 nmol/s/10(6) cells; mean ± standard deviation; n = 3). The (1) H NMR assay has adequate sensitivity to measure real-time pyruvate-lactate exchange kinetics in vitro, offering a complementary and accessible assay of apparent LDH activity.

  14. 129Xe chemical shift in human blood and pulmonary blood oxygenation measurement in humans using hyperpolarized 129Xe NMR

    PubMed Central

    Norquay, Graham; Leung, General; Stewart, Neil J.; Wolber, Jan

    2016-01-01

    Purpose To evaluate the dependency of the 129Xe‐red blood cell (RBC) chemical shift on blood oxygenation, and to use this relation for noninvasive measurement of pulmonary blood oxygenation in vivo with hyperpolarized 129Xe NMR. Methods Hyperpolarized 129Xe was equilibrated with blood samples of varying oxygenation in vitro, and NMR was performed at 1.5 T and 3 T. Dynamic in vivo NMR during breath hold apnea was performed at 3 T on two healthy volunteers following inhalation of hyperpolarized 129Xe. Results The 129Xe chemical shift in RBCs was found to increase nonlinearly with blood oxygenation at 1.5 T and 3 T. During breath hold apnea, the 129Xe chemical shift in RBCs exhibited a periodic time modulation and showed a net decrease in chemical shift of ∼1 ppm over a 35 s breath hold, corresponding to a decrease of 7–10 % in RBC oxygenation. The 129Xe‐RBC signal amplitude showed a modulation with the same frequency as the 129Xe‐RBC chemical shift. Conclusion The feasibility of using the 129Xe‐RBC chemical shift to measure pulmonary blood oxygenation in vivo has been demonstrated. Correlation between 129Xe‐RBC signal and 129Xe‐RBC chemical shift modulations in the lung warrants further investigation, with the aim to better quantify temporal blood oxygenation changes in the cardiopulmonary vascular circuit. Magn Reson Med 77:1399–1408, 2017. © 2016 The Authors Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. PMID:27062652

  15. Hyperpolarization-Activated Current (Ih) Is Reduced in Hippocampal Neurons from Gabra5−/− Mice

    PubMed Central

    Bonin, Robert P.; Zurek, Agnieszka A.; Yu, Jieying; Bayliss, Douglas A.; Orser, Beverley A.

    2013-01-01

    Changes in the expression of γ-aminobutyric acid type A (GABAA) receptors can either drive or mediate homeostatic alterations in neuronal excitability. A homeostatic relationship between α5 subunit-containing GABAA (α5GABAA) receptors that generate a tonic inhibitory conductance, and HCN channels that generate a hyperpolarization-activated cation current (Ih) was recently described for cortical neurons, where a reduction in Ih was accompanied by a reciprocal increase in the expression of α5GABAA receptors resulting in the preservation of dendritosomatic synaptic function. Here, we report that in mice that lack the α5 subunit gene (Gabra5−/−), cultured embryonic hippocampal pyramidal neurons and ex vivo CA1 hippocampal neurons unexpectedly exhibited a decrease in Ih current density (by 40% and 28%, respectively), compared with neurons from wild-type (WT) mice. The resting membrane potential and membrane hyperpolarization induced by blockade of Ih with ZD-7288 were similar in cultured WT and Gabra5−/− neurons. In contrast, membrane hyperpolarization measured after a train of action potentials was lower in Gabra5−/− neurons than in WT neurons. Also, membrane impedance measured in response to low frequency stimulation was greater in cultured Gabra5−/− neurons. Finally, the expression of HCN1 protein that generates Ih was reduced by 41% in the hippocampus of Gabra5−/− mice. These data indicate that loss of a tonic GABAergic inhibitory conductance was followed by a compensatory reduction in Ih. The results further suggest that the maintenance of resting membrane potential is preferentially maintained in mature and immature hippocampal neurons through the homeostatic co-regulation of structurally and biophysically distinct cation and anion channels. PMID:23516534

  16. Modulation of the hyperpolarization-activated Cl- current in human intestinal T84 epithelial cells by phosphorylation.

    PubMed Central

    Fritsch, J; Edelman, A

    1996-01-01

    1. Hyperpolarization-activated Cl- currents (ICl,hyp) were investigated in the T84 human adenocarcinoma cell line, using the patch-clamp whole-cell configuration. 2. During whole-cell recording with high-chloride and ATP-containing internal solutions, hyperpolarizing jumps from a holding potential of 0 mV elicited slow inward current relaxations, carried by Cl- and detected at membrane potentials more negative than -40 mV. Analysis of the relative permeabilities to monovalent anions gave the following sequence: Cl- > Br- > I- > glutamate. 3. ICl,hyp was partially inhibited by 1 mM diphenylamine-2-carboxylic acid or 0.1 mM 5-nitro-2-(3-phenylpropylamino)-benzoate, and was completely blocked by Cd2+ (> 300 microM). It was insensitive to 1 mM external 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid or 1 mM Ba2+. 4. ICl,hyp was inhibited by external application of 500 microM cptcAMP (8-(4-chlorophenylthio)-adenosine 3':5'-cyclic monophosphate) or 500 nM of the protein kinase C activator, phorbol 12-myristate, 13-acetate. 5. (i) Omission of ATP from the pipette solution, (ii) ATP replacement by the non-hydrolysable ATP analogue 5'-adenylylimidodiphosphate, and (iii) inhibition of protein kinase C by staurosporine or calphostin C accelerated the activation kinetics of the current and increased its amplitude, but did not alter its pharmacological properties. 6. We conclude that hyperpolarization-activated Cl- channels similar to those of ClC-2 channels (mammalian homologue of Torpedo chloride channel ClC-0) are present in T84 cells, and that their gating properties are modulated by phosphorylation. PMID:8745282

  17. Quantitation of a spin polarization-induced nuclear Overhauser effect (SPINOE) between a hyperpolarized (13) C-labeled cell metabolite and water protons.

    PubMed

    Marco-Rius, Irene; Bohndiek, Sarah E; Kettunen, Mikko I; Larkin, Timothy J; Basharat, Meer; Seeley, Colm; Brindle, Kevin M

    2014-01-01

    The spin polarization-induced nuclear Overhauser effect (SPINOE) describes the enhancement of spin polarization of solvent nuclei by the hyperpolarized spins of a solute. In this communication we demonstrate that SPINOEs can be observed between [1,4-(13) C2 ]fumarate, hyperpolarized using the dissolution dynamic nuclear polarization technique, and solvent water protons. We derive a theoretical expression for the expected enhancement and demonstrate that this fits well with experimental measurements. Although the magnitude of the effect is relatively small (around 2% measured here), the SPINOE increases at lower field strengths, so that at clinically relevant magnetic fields (1.5-3 T) it may be possible to track the passage through the circulation of a bolus containing a hyperpolarized (13) C-labeled substrate through the increase in solvent water (1) H signal.

  18. A Probabilistic Approach of Hazard Mapping for flow-type phenomena. An example of application at Mt. Etna

    NASA Astrophysics Data System (ADS)

    Bongolan, Vena Pearl; Iovine, Giulio; D'Ambrosio, Donato; Rongo, Rocco; Spataro, William

    2014-05-01

    The hazard induced by dangerous flow-type phenomena like lava flows, debris flows and debris avalanches can be usefully assessed by analysing a proper set of simulations of hypothetical events characterized in probability. In particular, to map lava-flow hazard, simulations can be performed by assuming a number of nodes from a regular grid of potential vents, selected to uniformly covering the study area. A probability of occurrence can then be assigned to each simulation, based on statistics of historical events and location of each vent with respect to the volcano. In this study, different hazard scenarios for Mt. Etna (Italy) have been realized, based on computer simulations of lava flows generated by a non-uniform grid of sources. The adopted grid covers the volcano with a variable density of nodes. Five macro-areas can in fact be recognised, in which at higher densities of nodes correspond higher probabilities of vent activation. Moreover, 4 distinct temporal frames have been considered (next 1, 25, 50 and 100 years) and related hazard scenarios have been computed. In addition, the topographic effects of the expected simulation within the considered temporal frames have been analysed, aiming at evaluating hazard trends due to natural morphological changes. Model parameters, e.g. the probability distribution function for vent activation and for types of eruption (distinguished into classes by duration and volume), have been derived by analysing the past 400-years volcanic history at Mt. Etna. Probabilities of vent activation for the 4 considered scenarios have been computed in terms of total number of expected events per each temporal frame. The actual number of events to be simulated per each scenario has been obtained by considering a Poisson distribution, with the number of expected events in that frame being the mean of the obtained probabilities. For each frame, a total of 240 runs have been performed. Each run is made of the set of simulations expected

  19. On the Use of Hyperpolarized Helium MRI for Conformal Avoidance Lung Radiotherapy

    SciTech Connect

    Hodge, C.W.; Tome, Wolfgang A.; Fain, S.B.; Bentzen, S.M.; Mehta, M.P.

    2010-01-01

    We wanted to illustrate the feasibility of using hyperpolarized helium magnetic resonance imaging (HPH-MRI) to obtain functional information that may assist in improving conformal avoidance of ventilating lung tissue during thoracic radiotherapy. HPH-MRI images were obtained from a volunteer patient and were first fused with a proton density-weighted (PD{sub w}) MRI to provide corresponding anatomic detail; they were then fused with the treatment planning computed tomography scan of a patient from our treatment planning database who possessed equivalent thoracic dimensions. An optimized treatment plan was then generated using the TomoTherapy treatment planning system, designating the HPH-enhancing regions as ventilation volume (VV). A dose-volume histogram compares the dosimetry of the lungs as a paired organ, the VV, and the lungs minus the VV. The clinical consequences of these changes was estimated using a bio-effect model, the parallel architecture model, or the local damage (f{sub dam}) model. Model parameters were chosen from published studies linking the incidence of grade 3+ pneumonitis, with the dose and volume irradiated. For two hypothetical treatment plans of 60 Gy in 30 fractions delivered to a right upper-lobe lung mass, one using and one ignoring the VV as an avoidance structure, the mean normalized total dose (NTD{sub mean}) values for the lung subvolumes were: lungs = 12.5 Gy{sub 3}vs. 13.52 Gy{sub 3}, VV = 9.94 Gy{sub 3}vs. 13.95 Gy{sub 3}, and lungs minus VV = 16.69 Gy{sub 3}vs. 19.16 Gy{sub 3}. Using the f{sub dam} values generated from these plans, one would predict a reduction of the incidence of grade 3+ radiation pneumonitis from 12%-4% when compared with a conventionally optimized plan. The use of HPH-MRI to identify ventilated lung subvolumes is feasible and has the potential to be incorporated into conformal avoidance treatment planning paradigms. A prospective clinical study evaluating this imaging technique is being developed.

  20. Pharmacological and physiological properties of the after-hyperpolarization current of bullfrog ganglion neurones.

    PubMed Central

    Goh, J W; Pennefather, P S

    1987-01-01

    1. The slowly decaying, calcium-dependent after-hyperpolarization (a.h.p.) that follows action potentials in bullfrog ganglion B cells has previously been shown to be generated by a potassium current called IAHP. We have recorded IAHP using a switched, single-electrode hybrid clamp where current-clamp mode was changed to voltage-clamp mode immediately after repolarization of a spike or the last spike of a train. 2. Reduction of extracellular calcium reduced the decay time of IAHP following a single spike. At all levels of extracellular calcium tested (0.5-4 mM), the decay time of IAHP was longer following a train of action potentials than following a single action potential. Thus, the time course of IAHP evoked by action potentials is a function of the calcium load induced by the action potentials. Conversely, agents that reduce the amount of IAHP activated without affecting its rate of decay, probably do not affect calcium influx. 3. Muscarine (2 or 10 microM) inhibits IAHP following an action potential by at most 30% and has no effect on decay rate of IAHP. These results suggest that muscarine has little or no effect on either calcium influx or sequestration. Decay of the a.h.p. is accelerated by muscarine but this effect is due to an increased leak conductance. 4. Charybdotoxin (CTX) between 4 and 20 nM, prolongs action potential duration in a manner consistent with blockade of the voltage- and calcium-dependent potassium current (Ic) involved in spike repolarization in these cells. This action is consistent with its reported action on analogous channels in other systems. However, CTX also reduces IAHP. Thus, in bullfrog ganglion neurones, two distinct calcium-dependent potassium currents exhibit a comparable sensitivity to CTX. This cannot be due to a decreased influx of calcium because the decay rate of IAHP following an action potential is unchanged. The action of CTX was observed with both crude and purified preparations of CTX. 5. Apamin (25 nM) and

  1. Developing hyperpolarized krypton-83 for nuclear magnetic resonance spectroscopy and magnetic resonance imaging

    NASA Astrophysics Data System (ADS)

    Cleveland, Zackary I.

    This dissertation discusses the production of highly nonequilibrium nuclear spin polarization, referred to as hyperpolarization or hp, in the nuclear spin I = 9/2 noble gas isotope krypton-83 using spin exchange optical pumping (SEOP). This nonequilibrium polarization yields nuclear magnetic resonance (NMR) signals that are enhanced three or more orders of magnitude above those of thermally polarized krypton and enables experiments that would otherwise be impossible. Krypton-83 possesses a nuclear electric quadrupole moment that dominates the longitudinal (T1) relaxation due to coupling of the quadrupole moment to fluctuating electric field gradients generated by distortions to the spherical symmetry of the electronic environment. Relaxation slows polarization buildup and limits the maximum signal intensity but makes krypton-83 a sensitive probe of its environment. The gas-phase krypton-83 longitudinal relaxation rate increases linearly with total gas density due to binary collisions. Density independent relaxation, caused by the formation of krypton-krypton van der Waals molecules and surface adsorption, also contributes to the observed rate. Buffer gases suppress van der Waals molecule mediated relaxation by breaking apart the weakly bound krypton dimers. Surface relaxation is gas composition independent and therefore more difficult to suppress. However, this relaxation mechanism makes hp krypton-83 sensitive to important surface properties including surface-to-volume ratio, surface chemistry, and surface temperature. The presence of surfaces with high krypton adsorption affinities (i.e. hydrophobic surfaces) accelerates the relaxation times and can produce T1 contrast in hp krypton-83 magnetic resonance imaging (MRI). Tobacco smoke deposited on surfaces generates strong T1 contrast allowing the observation of smoke deposition with spatial resolution. Conversely, water adsorption on surfaces significantly lengths the T1 times due competitive surface adsorption

  2. Steady-state free precession with hyperpolarized 3He: Experiments and theory

    NASA Astrophysics Data System (ADS)

    Wild, Jim M.; Teh, Kevin; Woodhouse, Neil; Paley, Martyn N. J.; Fichele, Stan; de Zanche, Nicola; Kasuboski, Larry

    2006-11-01

    The magnetization response of hyperpolarized 3He gas to a steady-state free precession (SSFP) sequence was simulated using matrix product operators. The simulations included the effects of flip angle ( α), sequence timings, resonant frequency, gas diffusion coefficient, imaging gradients, T1 and T2. Experiments performed at 1.5 T, on gas phantoms and with healthy human subjects, confirm the predicted theory, and indicate increased SNR with SSFP through use of higher flip angles when compared to optimized spoiled gradient echo (SPGR). Simulations and experiments show some compromise to the SNR and some point spread function broadening at high α due to the incomplete refocusing of transverse magnetization, caused by diffusion dephasing from the readout gradient. Mixing of gas polarization levels by diffusion between slices is also identified as a source of signal loss in SSFP at higher α through incomplete refocusing. Nevertheless, in the sample experiments, a SSFP sequence with an optimized flip angle of α = 20°, and 128 sequential phase encoding views, showed a higher SNR when compared to SPGR ( α = 7.2°) with the same bandwidth. Some of the gas sample experiments demonstrated a transient signal response that deviates from theory in the initial phase. This was identified as being caused by radiation damping interactions between the large initial transverse magnetization and the high quality factor ( Q = 250) birdcage resonator. In 3He NMR experiments, performed without imaging gradients, diffusion dephasing can be mitigated, and the effective T2 is relatively long (⩾1 s). Under these circumstances the SSFP sequence behaves like a CPMG sequence with sin( α/2) weighting of SNR. Experiments and simulations were also performed to characterize the off-resonance behaviour of the SSFP HP 3He signal. Characteristic banding artifacts due to off-resonance harmonic beating were observed in some of the in vivo SSFP images, for instance in axial slices close to the

  3. Steady-state free precession with hyperpolarized 3He: experiments and theory.

    PubMed

    Wild, Jim M; Teh, Kevin; Woodhouse, Neil; Paley, Martyn N J; Fichele, Stan; de Zanche, Nicola; Kasuboski, Larry

    2006-11-01

    The magnetization response of hyperpolarized 3He gas to a steady-state free precession (SSFP) sequence was simulated using matrix product operators. The simulations included the effects of flip angle (alpha), sequence timings, resonant frequency, gas diffusion coefficient, imaging gradients, T1 and T2. Experiments performed at 1.5 T, on gas phantoms and with healthy human subjects, confirm the predicted theory, and indicate increased SNR with SSFP through use of higher flip angles when compared to optimized spoiled gradient echo (SPGR). Simulations and experiments show some compromise to the SNR and some point spread function broadening at high alpha due to the incomplete refocusing of transverse magnetization, caused by diffusion dephasing from the readout gradient. Mixing of gas polarization levels by diffusion between slices is also identified as a source of signal loss in SSFP at higher alpha through incomplete refocusing. Nevertheless, in the sample experiments, a SSFP sequence with an optimized flip angle of alpha=20 degrees, and 128 sequential phase encoding views, showed a higher SNR when compared to SPGR (alpha=7.2 degrees) with the same bandwidth. Some of the gas sample experiments demonstrated a transient signal response that deviates from theory in the initial phase. This was identified as being caused by radiation damping interactions between the large initial transverse magnetization and the high quality factor (Q=250) birdcage resonator. In 3He NMR experiments, performed without imaging gradients, diffusion dephasing can be mitigated, and the effective T2 is relatively long (1 s). Under these circumstances the SSFP sequence behaves like a CPMG sequence with sinalpha/2 weighting of SNR. Experiments and simulations were also performed to characterize the off-resonance behaviour of the SSFP HP 3He signal. Characteristic banding artifacts due to off-resonance harmonic beating were observed in some of the in vivo SSFP images, for instance in axial slices

  4. Pharmacological and physiological properties of the after-hyperpolarization current of bullfrog ganglion neurones.

    PubMed

    Goh, J W; Pennefather, P S

    1987-12-01

    1. The slowly decaying, calcium-dependent after-hyperpolarization (a.h.p.) that follows action potentials in bullfrog ganglion B cells has previously been shown to be generated by a potassium current called IAHP. We have recorded IAHP using a switched, single-electrode hybrid clamp where current-clamp mode was changed to voltage-clamp mode immediately after repolarization of a spike or the last spike of a train. 2. Reduction of extracellular calcium reduced the decay time of IAHP following a single spike. At all levels of extracellular calcium tested (0.5-4 mM), the decay time of IAHP was longer following a train of action potentials than following a single action potential. Thus, the time course of IAHP evoked by action potentials is a function of the calcium load induced by the action potentials. Conversely, agents that reduce the amount of IAHP activated without affecting its rate of decay, probably do not affect calcium influx. 3. Muscarine (2 or 10 microM) inhibits IAHP following an action potential by at most 30% and has no effect on decay rate of IAHP. These results suggest that muscarine has little or no effect on either calcium influx or sequestration. Decay of the a.h.p. is accelerated by muscarine but this effect is due to an increased leak conductance. 4. Charybdotoxin (CTX) between 4 and 20 nM, prolongs action potential duration in a manner consistent with blockade of the voltage- and calcium-dependent potassium current (Ic) involved in spike repolarization in these cells. This action is consistent with its reported action on analogous channels in other systems. However, CTX also reduces IAHP. Thus, in bullfrog ganglion neurones, two distinct calcium-dependent potassium currents exhibit a comparable sensitivity to CTX. This cannot be due to a decreased influx of calcium because the decay rate of IAHP following an action potential is unchanged. The action of CTX was observed with both crude and purified preparations of CTX. 5. Apamin (25 nM) and

  5. Developmental Febrile Seizures Modulate Hippocampal Gene Expression of Hyperpolarization-Activated Channels in an Isoform- and Cell-Specific Manner

    PubMed Central

    Brewster, Amy; Bender, Roland A.; Chen, Yuncai; Dube, Celine; Eghbal-Ahmadi, Mariam; Baram, Tallie Z.

    2012-01-01

    Febrile seizures, in addition to being the most common seizure type of the developing human, may contribute to the generation of subsequent limbic epilepsy. Our previous work has demonstrated that prolonged experimental febrile seizures in the immature rat model increased hippocampal excitability long term, enhancing susceptibility to future seizures. The mechanisms for these profound proepileptogenic changes did not require cell death and were associated with long-term slowed kinetics of the hyperpolarization-activated depolarizing current (IH). Here we show that these seizures modulate the expression of genes encoding this current, the hyperpolarization-activated, cyclic nucleotide-gated channels (HCNs): In CA1 neurons expressing multiple HCN isoforms, the seizures induced a coordinated reduction of HCN1 mRNA and enhancement of HCN2 expression, thus altering the neuronal HCN phenotype. The seizure-induced augmentation of HCN2 expression involved CA3 in addition to CA1, whereas for HCN4, mRNA expression was not changed by the seizures in either hippocampal region. This isoform- and region-specific transcriptional regulation of the HCNs required neuronal activity rather than hyperthermia alone, correlated with seizure duration, and favored the formation of slow-kinetics HCN2-encoded channels. In summary, these data demonstrate a novel, activity-dependent transcriptional regulation of HCN molecules by developmental seizures. These changes result in long-lasting alteration of the HCN phenotype of specific hippocampal neuronal populations, with profound consequences on the excitability of the hippocampal network. PMID:12040066

  6. 32-channel phased-array receive with asymmetric birdcage transmit coil for hyperpolarized xenon-129 lung imaging.

    PubMed

    Dregely, Isabel; Ruset, Iulian C; Wiggins, Graham; Mareyam, Azma; Mugler, John P; Altes, Talissa A; Meyer, Craig; Ruppert, Kai; Wald, Lawrence L; Hersman, F William

    2013-08-01

    Hyperpolarized xenon-129 has the potential to become a noninvasive contrast agent for lung MRI. In addition to its utility for imaging of ventilated airspaces, the property of xenon to dissolve in lung tissue and blood upon inhalation provides the opportunity to study gas exchange. Implementations of imaging protocols for obtaining regional parameters that exploit the dissolved phase are limited by the available signal-to-noise ratio, excitation homogeneity, and length of acquisition times. To address these challenges, a 32-channel receive-array coil complemented by an asymmetric birdcage transmit coil tuned to the hyperpolarized xenon-129 resonance at 3 T was developed. First results of spin-density imaging in healthy subjects and subjects with obstructive lung disease demonstrated the improvements in image quality by high-resolution ventilation images with high signal-to-noise ratio. Parallel imaging performance of the phased-array coil was demonstrated by acceleration factors up to three in 2D acquisitions and up to six in 3D acquisitions. Transmit-field maps showed a regional variation of only 8% across the whole lung. The newly developed phased-array receive coil with the birdcage transmit coil will lead to an improvement in existing imaging protocols, but moreover enable the development of new, functional lung imaging protocols based on the improvements in excitation homogeneity, signal-to-noise ratio, and acquisition speed.

  7. Nonlinear effects of hyperpolarizing shifts in activation of mutant NaV1.7 channels on resting membrane potential.

    PubMed

    Estacion, Mark; Waxman, Stephen G

    2017-02-01

    The Nav1.7 sodium channel is preferentially expressed within dorsal root ganglion (DRG) and sympathetic ganglion neurons. Gain-of-function mutations that cause the painful disorder inherited erythromelalgia (IEM) shift channel activation in a hyperpolarizing direction. When expressed within DRG neurons, these mutations produce a depolarization of resting membrane potential (RMP). The biophysical basis for the depolarized RMP has to date not been established. To explore the effect on RMP of the shift in activation associated with a prototypical IEM mutation (L858H), we used dynamic clamp models that represent graded shifts that fractionate the effect of the mutation on activation voltage-dependence. Dynamic clamp recording from DRG neurons using a before-and-after protocol for each cell made it possible, even in the presence of cell-to-cell variation in starting RMP, to assess the effects of these graded mutant models. Our results demonstrate a non-linear, progressively larger effect on RMP as the shift in activation voltage-dependence becomes more hyperpolarized. The observed differences in RMP were predicted by the "late" current of each mutant model. Since the depolarization of RMP imposed by IEM mutant channels is known, in itself, to produce hyperexcitability of DRG neurons, the development of pharmacological agents that normalize or partially normalize activation voltage-dependence of IEM mutant channels merits further study.

  8. Local and bulk 13C hyperpolarization in nitrogen-vacancy-centred diamonds at variable fields and orientations

    PubMed Central

    Álvarez, Gonzalo A.; Bretschneider, Christian O.; Fischer, Ran; London, Paz; Kanda, Hisao; Onoda, Shinobu; Isoya, Junichi; Gershoni, David; Frydman, Lucio

    2015-01-01

    Polarizing nuclear spins is of fundamental importance in biology, chemistry and physics. Methods for hyperpolarizing 13C nuclei from free electrons in bulk usually demand operation at cryogenic temperatures. Room temperature approaches targeting diamonds with nitrogen-vacancy centres could alleviate this need; however, hitherto proposed strategies lack generality as they demand stringent conditions on the strength and/or alignment of the magnetic field. We report here an approach for achieving efficient electron-13C spin-alignment transfers, compatible with a broad range of magnetic field strengths and field orientations with respect to the diamond crystal. This versatility results from combining coherent microwave- and incoherent laser-induced transitions between selected energy states of the coupled electron–nuclear spin manifold. 13C-detected nuclear magnetic resonance experiments demonstrate that this hyperpolarization can be transferred via first-shell or via distant 13Cs throughout the nuclear bulk ensemble. This method opens new perspectives for applications of diamond nitrogen-vacancy centres in nuclear magnetic resonance, and in quantum information processing. PMID:26404169

  9. Hyperpolarization and inhibition of contraction mediated by nitric oxide released from enteric inhibitory neurones in guinea-pig taenia coli.

    PubMed Central

    Ward, S. M.; Dalziel, H. H.; Khoyi, M. A.; Westfall, A. S.; Sanders, K. M.; Westfall, D. P.

    1996-01-01

    1. Inhibition of nitric oxide synthase by NG-nitro-L-arginine (L-NNA) reduced the neurogenic relaxation of precontracted taenia coli only in the absence of atropine. The membrane hyperpolarization associated with the neurogenic relaxation was also reduced by inhibition of NOS only when atropine was absent. 2. The membrane hyperpolarization associated with the neurogenic relaxation of the taenia coli was inhibited by oxyhaemoglobin only in the absence of atropine. In the presence of atropine, oxyhaemoglobin did not reduce the i.j.p. or nerve evoked relaxation. 3. Inhibition of NOS by L-NNA did not affect the overflow of [3H]-ACh in response to electrical field stimulation (EFS), suggesting that, under the conditions of our experiments, endogenous NO did not modulate release of ACh. Sodium nitroprusside also had no effect on the neurogenic overflow of [3H]-ACh; however, noradrenaline significantly reduced [3H]-ACh overflow. 4. In summary, the postjunctional effects of neurally-released NO are not apparent in guinea-pig taenia coli when atropine is present. This implies muscarinic regulation of NO release or muscarinic regulation of another excitatory substance, such as tachykinin(s), that, when blocked, masks the postjunctional effects of NO. These data, together with previous studies, suggest a possible regulatory role for NO in enteric neurotransmission that may be more prominent in some species or tissues than others. PMID:8733575

  10. Rundown of the hyperpolarization-activated KAT1 channel involves slowing of the opening transitions regulated by phosphorylation.

    PubMed Central

    Tang, X D; Hoshi, T

    1999-01-01

    Disappearance of the functional activity or rundown of ion channels upon patch excision in many cells involves a decrease in the number of channels available to open. A variety of cellular and biophysical mechanisms have been shown to be involved in the rundown of different ion channels. We examined the rundown process of the plant hyperpolarization-activated KAT1 K+ channel expressed in Xenopus oocytes. The decrease in the KAT1 channel activity on patch excision was accompanied by progressive slowing of the activation time course, and it was caused by a shift in the voltage dependence of the channel without any change in the single-channel amplitude. The single-channel analysis showed that patch excision alters only the transitions leading up to the burst states of the channel. Patch cramming or concurrent application of protein kinase A (PKA) and ATP restored the channel activity. In contrast, nonspecific alkaline phosphatase (ALP) accelerated the rundown time course. Low internal pH, which inhibits ALP activity, slowed the KAT1 rundown time course. The results show that the opening transitions of the KAT1 channel are enhanced not only by hyperpolarization but also by PKA-mediated phosphorylation. PMID:10354434

  11. Reduced Hyperpolarization-Activated Current Contributes to Enhanced Intrinsic Excitability in Cultured Hippocampal Neurons from PrP−/− Mice

    PubMed Central

    Fan, Jing; Stemkowski, Patrick L.; Gandini, Maria A.; Black, Stefanie A.; Zhang, Zizhen; Souza, Ivana A.; Chen, Lina; Zamponi, Gerald W.

    2016-01-01

    Genetic ablation of cellular prion protein (PrPC) has been linked to increased neuronal excitability and synaptic activity in the hippocampus. We have previously shown that synaptic activity in hippocampi of PrP-null mice is increased due to enhanced N-methyl-D-aspartate receptor (NMDAR) function. Here, we focused on the effect of PRNP gene knock-out (KO) on intrinsic neuronal excitability, and in particular, the underlying ionic mechanism in hippocampal neurons cultured from P0 mouse pups. We found that the absence of PrPC profoundly affected the firing properties of cultured hippocampal neurons in the presence of synaptic blockers. The membrane impedance was greater in PrP-null neurons, and this difference was abolished by the hyperpolarization-activated cyclic nucleotide-gated (HCN) channel blocker ZD7288 (100 μM). HCN channel activity appeared to be functionally regulated by PrPC. The amplitude of voltage sag, a characteristic of activating HCN channel current (Ih), was decreased in null mice. Moreover, Ih peak current was reduced, along with a hyperpolarizing shift in activation gating and slower kinetics. However, neither HCN1 nor HCN2 formed a biochemical complex with PrPC. These results suggest that the absence of PrP downregulates the activity of HCN channels through activation of a cell signaling pathway rather than through direct interactions. This in turn contributes to an increase in membrane impedance to potentiate neuronal excitability. PMID:27047338

  12. Developmental febrile seizures modulate hippocampal gene expression of hyperpolarization-activated channels in an isoform- and cell-specific manner.

    PubMed

    Brewster, Amy; Bender, Roland A; Chen, Yuncai; Dube, Celine; Eghbal-Ahmadi, Mariam; Baram, Tallie Z

    2002-06-01

    Febrile seizures, in addition to being the most common seizure type of the developing human, may contribute to the generation of subsequent limbic epilepsy. Our previous work has demonstrated that prolonged experimental febrile seizures in the immature rat model increased hippocampal excitability long term, enhancing susceptibility to future seizures. The mechanisms for these profound proepileptogenic changes did not require cell death and were associated with long-term slowed kinetics of the hyperpolarization-activated depolarizing current (I(H)). Here we show that these seizures modulate the expression of genes encoding this current, the hyperpolarization-activated, cyclic nucleotide-gated channels (HCNs): In CA1 neurons expressing multiple HCN isoforms, the seizures induced a coordinated reduction of HCN1 mRNA and enhancement of HCN2 expression, thus altering the neuronal HCN phenotype. The seizure-induced augmentation of HCN2 expression involved CA3 in addition to CA1, whereas for HCN4, mRNA expression was not changed by the seizures in either hippocampal region. This isoform- and region-specific transcriptional regulation of the HCNs required neuronal activity rather than hyperthermia alone, correlated with seizure duration, and favored the formation of slow-kinetics HCN2-encoded channels. In summary, these data demonstrate a novel, activity-dependent transcriptional regulation of HCN molecules by developmental seizures. These changes result in long-lasting alteration of the HCN phenotype of specific hippocampal neuronal populations, with profound consequences on the excitability of the hippocampal network.

  13. Flow-induced activation of TRPV5 and TRPV6 channels stimulates Ca(2+)-activated K(+) channel causing membrane hyperpolarization.

    PubMed

    Cha, Seung-Kuy; Kim, Ji-Hee; Huang, Chou-Long

    2013-12-01

    TRPV5 and TRPV6 channels are expressed in distal renal tubules and play important roles in the transcellular Ca(2+) reabsorption in kidney. They are regulated by multiple intracellular factors including protein kinases A and C, membrane phospholipid PIP2, protons, and divalent ions Ca(2+) and Mg(2+). Here, we report that fluid flow that generates shear force within the physiological range of distal tubular fluid flow activated TRPV5 and TRPV6 channels expressed in HEK cells. Flow-induced activation of channel activity was reversible and did not desensitize over 2min. Fluid flow stimulated TRPV5 and 6-mediated Ca(2+) entry and increased intracellular Ca(2+) concentration. N-glycosylation-deficient TRPV5 channel was relatively insensitive to fluid flow. In cells coexpressing TRPV5 (or TRPV6) and Slo1-encoded maxi-K channels, fluid flow induced membrane hyperpolarization, which could be prevented by the maxi-K blocker iberiotoxin or TRPV5 and 6 blocker La(3+). In contrast, fluid flow did not cause membrane hyperpolarization in cells coexpressing ROMK1 and TRPV5 or 6 channel. These results reveal a new mechanism for the regulation of TRPV5 and TRPV6 channels. Activation of TRPV5 and TRPV6 by fluid flow may play a role in the regulation of flow-stimulated K(+) secretion via maxi-K channels in distal renal tubules and in the mechanism of pathogenesis of thiazide-induced hypocalciuria.

  14. Investigation of metabolic changes in STZ-induced diabetic rats with hyperpolarized [1-13C]acetate

    PubMed Central

    Koellisch, Ulrich; Laustsen, Christoffer; Nørlinger, Thomas S; Østergaard, Jakob Appel; Flyvbjerg, Allan; Gringeri, Concetta V; Menzel, Marion I; Schulte, Rolf F; Haase, Axel; Stødkilde-Jørgensen, Hans

    2015-01-01

    In the metabolism of acetate several enzymes are involved, which play an important role in free fatty acid oxidation. Fatty acid metabolism is altered in diabetes patients and therefore acetate might serve as a marker for pathological changes in the fuel selection of cells, as these changes occur in diabetes patients. Acetylcarnitine is a metabolic product of acetate, which enables its transport into the mitochondria for energy production. This study investigates whether the ratio of acetylcarnitine to acetate, measured by noninvasive hyperpolarized [1-13C]acetate magnetic resonance spectroscopy, could serve as a marker for myocardial, hepatic, and renal metabolic changes in rats with Streptozotocin (STZ)-induced diabetes in vivo. We demonstrate that the conversion of acetate to acetylcarnitine could be detected and quantified in all three organs of interest. More interestingly, we found that the hyperpolarized acetylcarnitine to acetate ratio was independent of blood glucose levels and prolonged hyperglycemia following diabetes induction in a type-1 diabetes model. PMID:26272734

  15. HIV-1 Tat Activates Neuronal Ryanodine Receptors with Rapid Induction of the Unfolded Protein Response and Mitochondrial Hyperpolarization

    PubMed Central

    Norman, John P.; Perry, Seth W.; Reynolds, Holly M.; Kiebala, Michelle; De Mesy Bentley, Karen L.; Trejo, Margarita; Volsky, David J.; Maggirwar, Sanjay B.; Dewhurst, Stephen; Masliah, Eliezer; Gelbard, Harris A.

    2008-01-01

    Neurologic disease caused by human immunodeficiency virus type 1 (HIV-1) is ultimately refractory to highly active antiretroviral therapy (HAART) because of failure of complete virus eradication in the central nervous system (CNS), and disruption of normal neural signaling events by virally induced chronic neuroinflammation. We have previously reported that HIV-1 Tat can induce mitochondrial hyperpolarization in cortical neurons, thus compromising the ability of the neuron to buffer calcium and sustain energy production for normal synaptic communication. In this report, we demonstrate that Tat induces rapid loss of ER calcium mediated by the ryanodine receptor (RyR), followed by the unfolded protein response (UPR) and pathologic dilatation of the ER in cortical neurons in vitro. RyR antagonism attenuated both Tat-mediated mitochondrial hyperpolarization and UPR induction. Delivery of Tat to murine CNS in vivo also leads to long-lasting pathologic ER dilatation and mitochondrial morphologic abnormalities. Finally, we performed ultrastructural studies that demonstrated mitochondria with abnormal morphology and dilated endoplasmic reticulum (ER) in brain tissue of patients with HIV-1 inflammation and neurodegeneration. Collectively, these data suggest that abnormal RyR signaling mediates the neuronal UPR with failure of mitochondrial energy metabolism, and is a critical locus for the neuropathogenesis of HIV-1 in the CNS. PMID:19009018

  16. Flow-Induced Activation of TRPV5 and TRPV6 Channel Stimulates Ca2+-Activated K+ Channel Causing Membrane Hyperpolarization

    PubMed Central

    Cha, Seung-Kuy; Kim, Ji-Hee; Huang, Chou-Long

    2014-01-01

    TRPV5 and TRPV6 channels are expressed in distal renal tubules and play important roles in the transcellular Ca2+ reabsorption in kidney. They are regulated by multiple intracellular factors including protein kinase A and C, membrane phospholipid PIP2, protons, and divalent ions Ca2+ and Mg2+. Here, we report that fluid flow that generates shear force within the physiological range of distal tubular fluid flow activated TRPV5 and TRPV6 channels expressed in HEK cells. Flow-induced activation of channel activity was reversible and did not desensitize over 2 minutes. Fluid flow stimulated TRPV5 and 6-mediated Ca2+ entry and increased intracellular Ca2+ concentration. N-glycosylation-deficient TRPV5 channel was relatively insensitive to fluid flow. In cells coexpressing TRPV5 (or TRPV6) and Slo1-encoded maxi-K channels, fluid flow induced membrane hyperpolarization, which could be prevented by the maxi-K blocker iberiotoxin or TRPV5 and 6 blocker La3+. In contrast, fluid flow did not cause membrane hyperpolarization in cells coexpressing ROMK1 and TRPV5 or 6 channels. These results reveal a new mechanism for regulation of TRPV5 and TRPV6 channels. Activation of TRPV5 and TRPV6 by fluid flow may play a role in the regulation of flow-stimulated K+ secretion via maxi-K channels in distal renal tubules and in the mechanism of pathogenesis of thiazide-induced hypocalciuria. PMID:24001793

  17. Differential effects of ascorbate on endothelium-derived hyperpolarizing factor (EDHF)-mediated vasodilatation in the bovine ciliary vascular bed and coronary artery.

    PubMed

    McNeish, Alister J; Nelli, Silvia; Wilson, William S; Dowell, Fiona J; Martin, William

    2003-03-01

    1. The ability of ascorbate to inhibit endothelium-derived hyperpolarizing factor (EDHF)-mediated vasodilatation was compared in the bovine perfused ciliary vascular bed and isolated rings of coronary artery. 2. Acetylcholine-induced, EDHF-mediated vasodilatation of the ciliary circulation was blocked following inclusion of ascorbate (50 micro M, 120 min) in the perfusion fluid. The blockade was highly selective since ascorbate had no effect on the vasodilator actions of the K(ATP) channel opener, levcromakalim, nor on the tonic vasodepressor action of basally released nitric oxide. 3. The possibility that concentration of ascorbate by the ciliary body was a prerequisite for blockade to occur was ruled out, since EDHF was still blocked when the anterior and posterior chambers were continuously flushed with Krebs solution or when both the aqueous and vitreous humour were drained. 4. Ascorbate at 50 micro M failed to affect bradykinin- or acetylcholine-induced, EDHF-mediated vasodilatation in rings of bovine coronary artery. Raising the concentration to 3 mM did produce blockade of EDHF, but this was nonselective, since vasodilator responses to endothelium-derived nitric oxide were also inhibited. 5. Thus, ascorbate (50 micro M) is not a universal blocker of EDHF. Whether its ability to block in the bovine ciliary circulation, but not in the coronary artery, is due to differences in the nature of EDHF at the two sites, differences in vessel size (resistance arterioles versus conduit artery), the presence or absence of flow, or to some other factor remains to be determined.

  18. Tonic nanomolar dopamine enables an activity-dependent phase recovery mechanism that persistently alters the maximal conductance of the hyperpolarization-activated current in a rhythmically active neuron.

    PubMed

    Rodgers, Edmund W; Fu, Jing Jing; Krenz, Wulf-Dieter C; Baro, Deborah J

    2011-11-09

    The phases at which network neurons fire in rhythmic motor outputs are critically important for the proper generation of motor behaviors. The pyloric network in the crustacean stomatogastric ganglion generates a rhythmic motor output wherein neuronal phase relationships are remarkably invariant across individuals and throughout lifetimes. The mechanisms for maintaining these robust phase relationships over the long-term are not well described. Here we show that tonic nanomolar dopamine (DA) acts at type 1 DA receptors (D1Rs) to enable an activity-dependent mechanism that can contribute to phase maintenance in the lateral pyloric (LP) neuron. The LP displays continuous rhythmic bursting. The activity-dependent mechanism was triggered by a prolonged decrease in LP burst duration, and it generated a persistent increase in the maximal conductance (G(max)) of the LP hyperpolarization-activated current (I(h)), but only in the presence of steady-state DA. Interestingly, micromolar DA produces an LP phase advance accompanied by a decrease in LP burst duration that abolishes normal LP network function. During a 1 h application of micromolar DA, LP phase recovered over tens of minutes because, the activity-dependent mechanism enabled by steady-state DA was triggered by the micromolar DA-induced decrease in LP burst duration. Presumably, this mechanism restored normal LP network function. These data suggest steady-state DA may enable homeostatic mechanisms that maintain motor network output during protracted neuromodulation. This DA-enabled, activity-dependent mechanism to preserve phase may be broadly relevant, as diminished dopaminergic tone has recently been shown to reduce I(h) in rhythmically active neurons in the mammalian brain.

  19. Tonic Nanomolar Dopamine Enables an Activity-Dependent Phase Recovery Mechanism That Persistently Alters the Maximal Conductance of the Hyperpolarization-Activated Current in a Rhythmically Active Neuron

    PubMed Central

    Rodgers, Edmund W.; Fu, Jing Jing; Krenz, Wulf-Dieter C.

    2011-01-01

    The phases at which network neurons fire in rhythmic motor outputs are critically important for the proper generation of motor behaviors. The pyloric network in the crustacean stomatogastric ganglion generates a rhythmic motor output wherein neuronal phase relationships are remarkably invariant across individuals and throughout lifetimes. The mechanisms for maintaining these robust phase relationships over the long-term are not well described. Here we show that tonic nanomolar dopamine (DA) acts at type 1 DA receptors (D1Rs) to enable an activity-dependent mechanism that can contribute to phase maintenance in the lateral pyloric (LP) neuron. The LP displays continuous rhythmic bursting. The activity-dependent mechanism was triggered by a prolonged decrease in LP burst duration, and it generated a persistent increase in the maximal conductance (Gmax) of the LP hyperpolarization-activated current (Ih), but only in the presence of steady-state DA. Interestingly, micromolar DA produces an LP phase advance accompanied by a decrease in LP burst duration that abolishes normal LP network function. During a 1 h application of micromolar DA, LP phase recovered over tens of minutes because, the activity-dependent mechanism enabled by steady-state DA was triggered by the micromolar DA-induced decrease in LP burst duration. Presumably, this mechanism restored normal LP network function. These data suggest steady-state DA may enable homeostatic mechanisms that maintain motor network output during protracted neuromodulation. This DA-enabled, activity-dependent mechanism to preserve phase may be broadly relevant, as diminished dopaminergic tone has recently been shown to reduce Ih in rhythmically active neurons in the mammalian brain. PMID:22072689

  20. Dynamic and High-Resolution Metabolic Imaging of Hyperpolarized [1-13C]-Pyruvate in the Rat Brain Using a High-Performance Gradient Insert

    PubMed Central

    Mayer, Dirk; Yen, Yi-Fen; Takahashi, Atsushi; Josan, Sonal; Tropp, James; Rutt, B.K.; Hurd, Ralph E.; Spielman, Daniel M.; Pfefferbaum, Adolf

    2010-01-01

    Fast chemical shift imaging techniques are advantageous in metabolic imaging of hyperpolarized compounds due to the limited duration of the signal amplification. At the same time, reducing the acquisition time in hyperpolarized imaging does not necessarily lead to the conventional penalty in signal-to-noise ratio that occurs in imaging at thermal equilibrium polarization levels. Here a high-performance gradient insert was used in combination with undersampled spiral chemical shift imaging to increase either the imaging speed or the spatial resolution of hyperpolarized 13C metabolic imaging on a clinical 3T MR scanner. Both a single-shot sequence with a total acquisition time of 125 ms and a 3-shot sequence with a nominal in-plane resolution of 1.5 mm were implemented. The k-space trajectories were measured and then used during image reconstruction. The technique was applied to metabolic imaging of the rat brain in vivo after the injection of hyperpolarized [1-13C]-pyruvate. Dynamic imaging afforded the measurement of region-of-interest-specific time courses of pyruvate and its metabolic products, while imaging at high spatial resolution was used to better characterize the spatial distribution of the metabolite signals. PMID:21500253

  1. Endothelium-dependent relaxation and hyperpolarization evoked by bradykinin in canine coronary arteries: enhancement by exercise-training.

    PubMed Central

    Mombouli, J. V.; Nakashima, M.; Hamra, M.; Vanhoutte, P. M.

    1996-01-01

    1. Kinins, which are produced locally in arterial walls, stimulate the release of endothelium-derived vasodilator substances. Therefore, they may participate in the metabolic adaptation to chronic exercise that occurs in the coronary circulation. Experiments were designed to compare the reactivity to bradykinin in coronary arteries isolated from sedentary and exercised-trained dogs (for 8-10 weeks). 2. The organ chambers used in this study were designed for measurement of isometric tension and cell membrane potential with glass microelectrodes. Rings of canine isolated coronary arteries with endothelium were suspended in the organ chambers filled with modified Krebs-Ringer bicarbonate solution (37 degrees C, gassed with 5% CO2 in 95 O2), and were all treated with indomethacin to prevent interference from prostaglandins. 3. Bradykinin evoked concentration-dependent relaxations of the coronary arteries. However, the kinin was significantly less potent in relaxing coronary arteries from the sedentary dogs than those from the trained ones. 4. In the presence of NG-nitro-L-arginine (an inhibitor of nitric oxide synthases), concentration-relaxation curves to bradykinin were shifted to the right in both types of preparations. Nonetheless, the peptide was still significantly more potent in arteries from exercise-trained animals. 5. In the electrophysiological experiments, concentration-hyperpolarization curves to bradykinin obtained in arteries from sedentary dogs were also significantly to the right of those in vessels from exercise-trained animals. Thus, in arteries from exercised animals, bradykinin more potently evoked the release of both nitric oxide (NO) and endothelium-derived hyperpolarizing factor (EDHF). 7. The angiotensin converting enzyme (ACE)-inhibitor, perindoprilat, shifted to the left the concentration-relaxation curves to bradykinin obtained under control conditions and in the presence of NG-nitro-L-arginine. The concentration-hyperpolarization curves to

  2. Hyperpolarization-activated current (I(h)) in vestibular calyx terminals: characterization and role in shaping postsynaptic events.

    PubMed

    Meredith, Frances L; Benke, Tim A; Rennie, Katherine J

    2012-12-01

    Calyx afferent terminals engulf the basolateral region of type I vestibular hair cells, and synaptic transmission across the vestibular type I hair cell/calyx is not well understood. Calyces express several ionic conductances, which may shape postsynaptic potentials. These include previously described tetrodotoxin-sensitive inward Na(+) currents, voltage-dependent outward K(+) currents and a K(Ca) current. Here, we characterize an inwardly rectifying conductance in gerbil semicircular canal calyx terminals (postnatal days 3-45), sensitive to voltage and to cyclic nucleotides. Using whole-cell patch clamp, we recorded from isolated calyx terminals still attached to their type I hair cells. A slowly activating, noninactivating current (I(h)) was seen with hyperpolarizing voltage steps negative to the resting potential. External Cs(+) (1-5 mM) and ZD7288 (100 μM) blocked the inward current by 97 and 83 %, respectively, confirming that I(h) was carried by hyperpolarization-activated, cyclic nucleotide gated channels. Mean half-activation voltage of I(h) was -123 mV, which shifted to -114 mV in the presence of cAMP. Activation of I(h) was well described with a third order exponential fit to the current (mean time constant of activation, τ, was 190 ms at -139 mV). Activation speeded up significantly (τ=136 and 127 ms, respectively) when intracellular cAMP and cGMP were present, suggesting that in vivo I(h) could be subject to efferent modulation via cyclic nucleotide-dependent mechanisms. In current clamp, hyperpolarizing current steps produced a time-dependent depolarizing sag followed by either a rebound afterdepolarization or an action potential. Spontaneous excitatory postsynaptic potentials (EPSPs) became larger and wider when I(h) was blocked with ZD7288. In a three-dimensional mathematical model of the calyx terminal based on Hodgkin-Huxley type ionic conductances, removal of I(h) similarly increased the EPSP, whereas cAMP slightly decreased simulated EPSP size

  3. GABA actions in hippocampal area CA3 during postnatal development: differential shift from depolarizing to hyperpolarizing in somatic and dendritic compartments.

    PubMed

    Romo-Parra, Héctor; Treviño, Mario; Heinemann, Uwe; Gutiérrez, Rafael

    2008-03-01

    Gamma-aminobutyric acid type A receptor (GABA(A)-R) activation leads to depolarization of pyramidal cells during the first postnatal week and produces hyperpolarization from the second week. However, immunohistochemical evidence has suggested that during the second and third postnatal weeks the NKCC1 cotransporter relocates from the soma to the dendrites of CA3 pyramidal cells. We hypothesized that this leads to depolarizing responses in apical dendrites. Here we show that the activation of GABA(A)-R in the distal dendrites of CA3 pyramidal cells at P15 by restricted application of muscimol or synaptic activation by stimulation of interneurons in stratum radiatum (SR) causes depolarizing postsynaptic potentials (PSPs), which are blocked by NKCC1 cotransporter antagonists. By contrast, activation of proximal GABA(A)-R by muscimol application or by stimulation of interneurons in s. oriens (SO) leads to hyperpolarizing PSPs. Activation of the dentate gyrus (DG) in the presence of glutamatergic blockers evokes hyperpolarizing responses during the second postnatal week; however, the reversal potential of the DG-evoked inhibitory (I)PSPs is more depolarized than that of IPSPs evoked by activation of SO interneurons. Despite the shift of GABA action from depolarizing to hyperpolarizing, DG-evoked field potentials (f-PSPs) recorded in s. lucidum/radiatum (SL/R) do not change in polarity until the third week. Current source density analysis yielded results consistent with depolarizing actions of GABA in the dendritic compartment. Our data suggest that GABAergic input to apical dendrites of pyramidal cells of CA3 evokes depolarizing PSPs long after synaptic inhibition has become hyperpolarizing in the somata, in the axon initial segments and in basal dendrites.

  4. The hyperpolarization-activated current (Ih) and its contribution to pacemaker activity in rat CA1 hippocampal stratum oriens-alveus interneurones.

    PubMed Central

    Maccaferri, G; McBain, C J

    1996-01-01

    1. The hyperpolarization-activated current (Ih) and its role in pacemaking activity in rat hippocampal stratum oriens-alveus interneurones was studied using whole-cell and perforated patch-clamp configurations. 2. Voltage-clamp recordings revealed Ih as a slowly activating, inward current, activated by hyperpolarizing steps (holding potential, Vh = -40 mV), with a reversal potential close to -30 mV. Its activation curve ranged from approximately -50 to -120 mV with a mid-activation point of -84.1 mV. 3. Ih was blocked by external application of Cs+ (2-5 mM) and ZD7288 (100 microM), but not by Ba2+ (1 mM). 4. Ih was potentiated by both noradrenaline and isoprenaline by a mechanism consistent with a shift in the Ih activation curve. 5. Under current-clamp conditions (Vh = -60 mV), ZD7288 induced a membrane hyperpolarization concomitant with an increase in the membrane input resistance and abolished the voltage sag generated by hyperpolarizing current injection. 6. Analysis of the current-discharge relationship revealed that block of Ih differentially increased the firing frequency of spikes occurring early in the train compared with those occurring late in the discharge. 7. When applied to spontaneously firing cells, ZD7288 reduced the firing frequency by selectively altering the time course of the interspike interval, while minimally affecting other action potential characteristics. Similarly, isoprenaline increased the spontaneous firing frequency by an effect exclusively on the after-hyperpolarization and interspike interval. 8. These results provide evidence for the involvement of Ih in the excitability and generation of spontaneous firing in hippocampal stratum oriens-alveus interneurones. PMID:8951716

  5. Rapid sequential injections of hyperpolarized [1-¹³C]pyruvate in vivo using a sub-kelvin, multi-sample DNP polarizer.

    PubMed

    Hu, Simon; Larson, Peder E Z; Vancriekinge, Mark; Leach, Andrew M; Park, Ilwoo; Leon, Christine; Zhou, Jenny; Shin, Peter J; Reed, Galen; Keselman, Paul; von Morze, Cornelius; Yoshihara, Hikari; Bok, Robert A; Nelson, Sarah J; Kurhanewicz, John; Vigneron, Daniel B

    2013-05-01

    The development of hyperpolarized technology utilizing dynamic nuclear polarization (DNP) has enabled the rapid measurement of (13)C metabolism in vivo with very high SNR. However, with traditional DNP equipment, consecutive injections of a hyperpolarized compound in an animal have been subject to a practical minimum time between injections governed by the polarization build-up time, which is on the order of an hour for [1-(13)C]pyruvate. This has precluded the monitoring of metabolic changes occurring on a faster time scale. In this study, we demonstrated the ability to acquire in vivo dynamic magnetic resonance spectroscopy (MRS) and 3D magnetic resonance spectroscopic imaging (MRSI) data in normal rats with a 5 min interval between injections of hyperpolarized [1-(13)C]pyruvate using a prototype, sub-Kelvin dynamic nuclear polarizer with the capability to simultaneously polarize up to 4 samples and dissolve them in rapid succession. There were minimal perturbations in the hyperpolarized spectra as a result of the multiple injections, suggesting that such an approach would not confound the investigation of metabolism occurring on this time scale. As an initial demonstration of the application of this technology and approach for monitoring rapid changes in metabolism as a result of a physiological intervention, we investigated the pharmacodynamics of the anti-cancer agent dichloroacetate (DCA), collecting hyperpolarized data before administration of DCA, 1 min after administration, and 6 min after administration. Dramatic increases in (13)C-bicarbonate were detected just 1 min (as well as 6 min) after DCA administration.

  6. Src Kinase Is the Connecting Player between Protein Kinase A (PKA) Activation and Hyperpolarization through SLO3 Potassium Channel Regulation in Mouse Sperm.

    PubMed

    Stival, Cintia; La Spina, Florenza A; Baró Graf, Carolina; Arcelay, Enid; Arranz, Silvia E; Ferreira, Juan J; Le Grand, Sibylle; Dzikunu, Victor A; Santi, Celia M; Visconti, Pablo E; Buffone, Mariano G; Krapf, Dario

    2015-07-24

    Plasma membrane hyperpolarization is crucial for mammalian sperm to acquire acrosomal responsiveness during capacitation. Among the signaling events leading to mammalian sperm capacitation, the immediate activation of protein kinase A plays a pivotal role, promoting the subsequent stimulation of protein tyrosine phosphorylation that associates with fertilizing capacity. We have shown previously that mice deficient in the tyrosine kinase cSrc are infertile and exhibit improper cauda epididymis development. It is therefore not clear whether lack of sperm functionality is due to problems in epididymal maturation or to the absence of cSrc in sperm. To further address this problem, we investigated the kinetics of cSrc activation using anti-Tyr(P)-416-cSrc antibodies that only recognize active cSrc. Our results provide evidence that cSrc is activated downstream of PKA and that inhibition of its activity blocks the capacitation-induced hyperpolarization of the sperm plasma membrane without blocking the increase in tyrosine phosphorylation that accompanies capacitation. In addition, we show that cSrc inhibition also blocks the agonist-induced acrosome reaction and that this inhibition is overcome by pharmacological hyperpolarization. Considering that capacitation-induced hyperpolarization is mediated by SLO3, we evaluated the action of cSrc inhibitors on the heterologously expressed SLO3 channel. Our results indicate that, similar to SLO1 K(+) channels, cSrc blockers significantly decreased SLO3-mediated currents. Together, these results are consistent with findings showing that hyperpolarization of the sperm plasma membrane is necessary and sufficient to prepare the sperm for the acrosome reaction and suggest that changes in sperm membrane potential are mediated by cSrc activation.

  7. The hyperpolarization-activated non-specific cation current (In ) adjusts the membrane properties, excitability, and activity pattern of the giant cells in the rat dorsal cochlear nucleus.

    PubMed

    Rusznák, Zoltán; Pál, Balázs; Kőszeghy, Aron; Fu, Yuhong; Szücs, Géza; Paxinos, George

    2013-03-01

    Giant cells of the cochlear nucleus are thought to integrate multimodal sensory inputs and participate in monaural sound source localization. Our aim was to explore the significance of a hyperpolarization-activated current in determining the activity of giant neurones in slices prepared from 10 to 14-day-old rats. When subjected to hyperpolarizing stimuli, giant cells produced a 4-(N-ethyl-N-phenylamino)-1,2-dimethyl-6-(methylamino) pyridinium chloride (ZD7288)-sensitive inward current with a reversal potential and half-activation voltage of -36 and -88 mV, respectively. Consequently, the current was identified as the hyperpolarization-activated non-specific cationic current (Ih ). At the resting membrane potential, 3.5% of the maximum Ih conductance was available. Immunohistochemistry experiments suggested that hyperpolarization-activated, cyclic nucleotide-gated, cation non-selective (HCN)1, HCN2, and HCN4 subunits contribute to the assembly of the functional channels. Inhibition of Ih hyperpolarized the membrane by 6 mV and impeded spontaneous firing. The frequencies of spontaneous inhibitory and excitatory postsynaptic currents reaching the giant cell bodies were reduced but no significant change was observed when evoked postsynaptic currents were recorded. Giant cells are affected by biphasic postsynaptic currents consisting of an excitatory and a subsequent inhibitory component. Inhibition of Ih reduced the frequency of these biphasic events by 65% and increased the decay time constants of the inhibitory component. We conclude that Ih adjusts the resting membrane potential, contributes to spontaneous action potential firing, and may participate in the dendritic integration of the synaptic inputs of the giant neurones. Because its amplitude was higher in young than in adult rats, Ih of the giant cells may be especially important during the postnatal maturation of the auditory system.

  8. Platelet-derived growth factor receptor-α-positive cells and not smooth muscle cells mediate purinergic hyperpolarization in murine colonic muscles.

    PubMed

    Kurahashi, Masaaki; Mutafova-Yambolieva, Violeta; Koh, Sang Don; Sanders, Kenton M

    2014-09-15

    Enteric inhibitory neurotransmission is an important feature of the neural regulation of gastrointestinal motility. Purinergic neurotransmission, via P2Y1 receptors, mediates one phase of inhibitory neural control. For decades, ATP has been assumed to be the purinergic neurotransmitter and smooth muscle cells (SMCs) have been considered the primary targets for inhibitory neurotransmission. Recent experiments have cast doubt on both of these assumptions and suggested that another cell type, platelet-derived growth factor receptor-α-positive (PDGFRα(+)) cells, is the target for purinergic neurotransmission. We compared responses of PDGFRα(+) cells and SMCs to several purine compounds to determine if these cells responded in a manner consistent with enteric inhibitory neurotransmission. ATP hyperpolarized PDGFRα(+) cells but depolarized SMCs. Only part of the ATP response in PDGFRα(+) cells was blocked by MRS 2500, a P2Y1 antagonist. ADP, MRS 2365, β-NAD, and adenosine 5-diphosphate-ribose, P2Y1 agonists, hyperpolarized PDGFRα(+) cells, and these responses were blocked by MRS 2500. Adenosine 5-diphosphate-ribose was more potent in eliciting hyperpolarization responses than β-NAD. P2Y1 agonists failed to elicit responses in SMCs. Small hyperpolarization responses were elicited in SMCs by a small-conductance Ca(2+)-activated K(+) channel agonist, cyclohexyl-[2-(3,5-dimethyl-pyrazol-1-yl)-6-methyl-pyrimidin-4-yl]-amine, consistent with the low expression and current density of small-conductance Ca(2+)-activated K(+) channels in these cells. Large-amplitude hyperpolarization responses, elicited in PDGFRα(+) cells, but not SMCs, by P2Y1 agonists are consistent with the generation of inhibitory junction potentials in intact muscles in response to purinergic neurotransmission. The responses of PDGFRα(+) cells and SMCs to purines suggest that SMCs are unlikely targets for purinergic neurotransmission in colonic muscles.

  9. Transfer of hyperpolarization from long T 1 storage nuclei to short T 1 neighbors using FLOPSY-8

    NASA Astrophysics Data System (ADS)

    Moreno, Karlos X.; Harrison, Crystal; Dean Sherry, A.; Malloy, Craig R.; Merritt, Matthew E.

    2011-12-01

    Nuclei with long T 1s are optimal targets for dynamic nuclear polarization (DNP). Therefore, most of the agents used in metabolic imaging and spectroscopy studies are based on carboxylic acid moieties that lack protons, a strong source of dipolar relaxation. Metabolic flux information encoded into spectra of small molecule metabolites in the form of the 13C isotopomer data cannot be accessed using standard 13C hyperpolarization methods because protonated carbons relax too quickly through T 1 dipolar relaxation. It is shown here that the longitudinal mixing sequence FLOPSY-8 can be used to transfer polarization from a long T 1 storage nucleus to adjacent protonated carbons so that they may be detected with high sensitivity. We demonstrate that FLOPSY-8 allows a direct readout of isotopomer populations in butyrate and glutamate in vitro.

  10. Effects of fasting on serial measurements of hyperpolarized [1-(13) C]pyruvate metabolism in tumors.

    PubMed

    Serrao, Eva M; Rodrigues, Tiago B; Gallagher, Ferdia A; Kettunen, Mikko I; Kennedy, Brett W C; Vowler, Sarah L; Burling, Keith A; Brindle, Kevin M

    2016-08-01

    Imaging of the metabolism of hyperpolarized [1-(13) C]pyruvate has shown considerable promise in preclinical studies in oncology, particularly for the assessment of early treatment response. The repeatability of measurements of (13) C label exchange between pyruvate and lactate was determined in a murine lymphoma model in fasted and non-fasted animals. The fasted state showed lower intra-individual variability, although the [1-(13) C]lactate/[1-(13) C]pyruvate signal ratio was significantly greater in fasted than in non-fasted mice, which may be explained by the higher tumor lactate concentrations in fasted animals. These results indicate that the fasted state may be preferable for the measurement of (13) C label exchange between pyruvate and lactate, as it reduces the variability and therefore should make it easier to detect the effects of therapy. © 2016 The Authors. NMR in Biomedicine published by John Wiley & Sons Ltd.

  11. Effects of fasting on serial measurements of hyperpolarized [1‐13C]pyruvate metabolism in tumors

    PubMed Central

    Serrao, Eva M.; Rodrigues, Tiago B.; Gallagher, Ferdia A.; Kettunen, Mikko I.; Kennedy, Brett W. C.; Vowler, Sarah L.; Burling, Keith A.

    2016-01-01

    Imaging of the metabolism of hyperpolarized [1‐13C]pyruvate has shown considerable promise in preclinical studies in oncology, particularly for the assessment of early treatment response. The repeatability of measurements of 13C label exchange between pyruvate and lactate was determined in a murine lymphoma model in fasted and non‐fasted animals. The fasted state showed lower intra‐individual variability, although the [1‐13C]lactate/[1‐13C]pyruvate signal ratio was significantly greater in fasted than in non‐fasted mice, which may be explained by the higher tumor lactate concentrations in fasted animals. These results indicate that the fasted state may be preferable for the measurement of 13C label exchange between pyruvate and lactate, as it reduces the variability and therefore should make it easier to detect the effects of therapy. © 2016 The Authors. NMR in Biomedicine published by John Wiley & Sons Ltd. PMID:27309986

  12. Characterization of the hyperpolarization-activated current, I(f), in ventricular myocytes isolated from hypertensive rats.

    PubMed Central

    Cerbai, E; Barbieri, M; Mugelli, A

    1994-01-01

    1. Left ventricular myocytes isolated from the heart of young (2-month-old) and old (18- to 20-month-old) spontaneously hypertensive rats (SHRs) were studied in the whole-cell configuration. Since multicellular preparations from old SHRs show a diastolic depolarization phase, we performed experiments to test whether it was associated with the presence of a hyperpolarization-activated If-like current. 2. In control Tyrode solution, a time-dependent increasing inward current activated by hyperpolarization was recorded in myocytes from old SHRs showing a diastolic depolarization phase. A barium-insensitive, caesium-sensitive, time-dependent inward current was recorded in a minority (4 of 33) of cells from young SHRs (membrane capacitance, 160 +/- 7 pF) but in 93% (25 of 27, P < 0.01) of myocytes from old SHRs (membrane capacitance, 355 +/- 19 pF, P < 0.01). 3. The current was fully activated at -120 mV and voltage of half-maximal activation was -88.1 +/- 1.5 mV; it was blocked by extracellular CsCl (4 mM) in a voltage-dependent manner. Reducing [K+]o from 25 to 5.4 mM caused a shift of the reversal potential from -17.3 +/- 3.8 to -25.7 +/- 2.7 mV and a 60% decrease of current conductance. 4. These findings suggest that an If-like current is present in rat ventricular myocytes from old SHRs, where it might favour the occurrence of spontaneous action potentials. PMID:7707227

  13. Alpha-2 noradrenergic receptor activation inhibits the hyperpolarization-activated cation current (Ih ) in neurons of the ventral tegmental area

    PubMed Central

    Inyushin, Mikhail U.; Arencibia-Albite, Francisco; Vázquez-Torres, Rafael; Vélez-Hernández, María E.; Jiménez-Rivera, Carlos A.

    2010-01-01

    The ventral tegmental area (VTA) is the source of dopaminergic projections innervating cortical structures and ventral forebrain. Dysfunction of this mesocorticolimbic system is critically involved in psychiatric disorders such as addiction and schizophrenia. Changes in VTA dopamine (DA) neuronal activity can alter neurotransmitter release at target regions which modify information processing in the reward circuit. Here we studied the effect of α-2 noradrenergic receptor activation on the hyperpolarization-activated cation current (Ih ) in DA neurons of the rat VTA. Brain slice preparations using whole-cell current and voltage-clamp techniques were employed. Clonidine and UK14304 (α-2 receptor selective agonists) were found to decrease Ih amplitude and to slow its rate of activation indicating a negative shift in the current’s voltage dependence. Two non-subtype-selective α-2 receptor antagonists, yohimbine and RS79948, prevented the effects of α-2 receptor activation. RX821002, a noradrenergic antagonist specific for α-2A and α-2D did not prevent Ih inhibition. This result suggests that clonidine might be acting via an α-2C subtype since this receptor is the most abundant variant in the VTA. Analysis of a second messenger system associated with the α-2 receptor revealed that Ih inhibition is independent of cyclic adenosine monophosphate (cAMP) and resulted from the activation of protein kinase C. It is suggested that the α-2 mediated hyperpolarizing shift in Ih voltage dependence can facilitate the transition from pacemaker firing to afferent-driven burst activity. This transition may play a key role on the changes in synaptic plasticity that occurs in the mesocorticolimbic system under pathological conditions. PMID:20122999

  14. Real-time Detection of Hepatic Gluconeogenic and Glycogenolytic States Using Hyperpolarized [2-13C]Dihydroxyacetone*

    PubMed Central

    Moreno, Karlos X.; Satapati, Santhosh; DeBerardinis, Ralph J.; Burgess, Shawn C.; Malloy, Craig R.; Merritt, Matthew E.

    2014-01-01

    Glycogenolysis and gluconeogenesis are sensitive to nutritional state, and the net direction of flux is controlled by multiple enzymatic steps. This delicate balance in the liver is disrupted by a variety of pathological states including cancer and diabetes mellitus. Hyperpolarized carbon-13 magnetic resonance is a new metabolic imaging technique that can probe intermediary metabolism nondestructively. There are currently no methods to rapidly distinguish livers in a gluconeogenic from glycogenolytic state. Here we use the gluconeogenic precursor dihydroxyacetone (DHA) to deliver hyperpolarized carbon-13 to the perfused mouse liver. DHA enters gluconeogenesis at the level of the trioses. Perfusion conditions were designed to establish either a gluconeogenic or a glycogenolytic state. Unexpectedly, we found that [2-13C]DHA was metabolized within a few seconds to the common intermediates and end products of both glycolysis and gluconeogenesis under both conditions, including [2,5-13C]glucose, [2-13C]glycerol 3-phosphate, [2-13C]phosphoenolpyruvate (PEP), [2-13C]pyruvate, [2-13C]alanine, and [2-13C]lactate. [2-13C]Phosphoenolpyruvate, a key branch point in gluconeogenesis and glycolysis, was monitored in functioning tissue for the first time. Observation of [2-13C]PEP was not anticipated as the free energy difference between PEP and pyruvate is large. Pyruvate kinase is the only regulatory step of the common glycolytic-gluconeogenic pathway that appears to exert significant control over the kinetics of any metabolites of DHA. A ratio of glycolytic to gluconeogenic products distinguished the gluconeogenic from glycogenolytic state in these functioning livers. PMID:25352600

  15. Comparison of Kinetic Models for Analysis of Pyruvate-to-Lactate Exchange by Hyperpolarized 13C NMR

    PubMed Central

    Harrison, Crystal; Yang, Chendong; Jindal, Ashish; DeBerardinis, Ralph J.; Hooshyar, M.A.; Merritt, Matthew; Sherry, A. Dean; Malloy, Craig R.

    2012-01-01

    The activity of specific enzyme-catalyzed reactions may be detected in vivo by 13C NMR of hyperpolarized (HP) substrates. The signal from HP substrates and products, acquired over time, have been fit to a number of different mathematical models to determine fluxes, but these models have not been critically compared. In this study, two-pool and three-pool first-order models were constructed to measure flux through lactate dehydrogenase in isolated glioblastoma cells by NMR detection of lactate and pyruvate following addition of hyperpolarized [1-13C]pyruvate. Mass spectrometry (MS) was used to independently monitor 13C enrichment in intra- and extracellular lactate. Six models were evaluated using time dependent pyruvate C2 and lactate C1 HP NMR data acquired by use of selective excitation pulses plus 13C enrichment data from intracellular and extracellular lactate measured by MS. A three-pool bi-directional model provided the most accurate description of pyruvate metabolism in these cells. With computed values for the T1 of pyruvate and lactate as well as the effect of pulsing, the initial flux through lactate dehydrogenase (LDH) was well-determined by both the two-pool bidirectional and unidirectional models when only HP data was available. The three-pool model was necessary to fit the combined data from both MS and HP, but the simpler two-pool exchange model was sufficient to determine the 13C lactate concentration when the lactate appearance was measured only by HP. PMID:22451442

  16. Real-time detection of hepatic gluconeogenic and glycogenolytic states using hyperpolarized [2-13C]dihydroxyacetone.

    PubMed

    Moreno, Karlos X; Satapati, Santhosh; DeBerardinis, Ralph J; Burgess, Shawn C; Malloy, Craig R; Merritt, Matthew E

    2014-12-26

    Glycogenolysis and gluconeogenesis are sensitive to nutritional state, and the net direction of flux is controlled by multiple enzymatic steps. This delicate balance in the liver is disrupted by a variety of pathological states including cancer and diabetes mellitus. Hyperpolarized carbon-13 magnetic resonance is a new metabolic imaging technique that can probe intermediary metabolism nondestructively. There are currently no methods to rapidly distinguish livers in a gluconeogenic from glycogenolytic state. Here we use the gluconeogenic precursor dihydroxyacetone (DHA) to deliver hyperpolarized carbon-13 to the perfused mouse liver. DHA enters gluconeogenesis at the level of the trioses. Perfusion conditions were designed to establish either a gluconeogenic or a glycogenolytic state. Unexpectedly, we found that [2-(13)C]DHA was metabolized within a few seconds to the common intermediates and end products of both glycolysis and gluconeogenesis under both conditions, including [2,5-(13)C]glucose, [2-(13)C]glycerol 3-phosphate, [2-(13)C]phosphoenolpyruvate (PEP), [2-(13)C]pyruvate, [2-(13)C]alanine, and [2-(13)C]lactate. [2-(13)C]Phosphoenolpyruvate, a key branch point in gluconeogenesis and glycolysis, was monitored in functioning tissue for the first time. Observation of [2-(13)C]PEP was not anticipated as the free energy difference between PEP and pyruvate is large. Pyruvate kinase is the only regulatory step of the common glycolytic-gluconeogenic pathway that appears to exert significant control over the kinetics of any metabolites of DHA. A ratio of glycolytic to gluconeogenic products distinguished the gluconeogenic from glycogenolytic state in these functioning livers.

  17. Cation-dependent gating of the hyperpolarization-activated cation current in the rabbit sino-atrial node cells.

    PubMed Central

    Maruoka, F; Nakashima, Y; Takano, M; Ono, K; Noma, A

    1994-01-01

    1. The gating properties of the hyperpolarization-activated cation current (I(f) or Ih) were investigated in single pacemaker cells dissociated from the rabbit sino-atrial node. 2. The whole-cell I(f) was recorded in the presence of different external cations. The inward I(f) was increased when external Na+ was replaced with K+, and was decreased in Li+ or Rb+ solution. In Tris+ and Cs+ solutions, the inward I(f) was negligible. The outward tail current recorded upon depolarization was largest in Li+ solution and smaller in a sequence of Na+, Tris+ and K+ solutions. In Rb+ and Cs+ solutions, only a small tail current was recorded. 3. The outward tail current had a 'shoulder' in Na+ solution, which was much delayed by replacing Na+ with Li+. In K+ solution, the decay of the tail current was much faster, and no obvious shoulder was recorded. The tail current was slowest in Li(+)-rich and 0 mM K+ solution, and was progressively accelerated by adding K+ over the range from 0 to 3 mM. The tail current at 30 mM [K+]o showed only a small shoulder. A common binding site to modulate the I(f) deactivation was suggested for monovalent cations. 4. The shoulder of the I(f) tail became more evident as I(f) was activated to a larger extent either by prolonging the duration or by increasing the amplitude of the preceding hyperpolarization in both Na+ and Li+ solutions. 5. The I(f) was first activated by hyperpolarizing the membrane to -110 mV, and then deactivated by depolarization. The inward tail current at -50 mV showed a single exponential decay. At more positive potentials, the shoulder of the outward tail currents became more evident and the rate of the final decay was increased. 6. The time course of I(f) activation was well fitted with the sum of two exponential functions. Time constants of both components were not affected by the external cation (Na+, K+ or Li+) replacement. Likewise, the quasi-steady state activation was conserved when external Na+ was replaced with Li

  18. High-Resolution 3D Proton MRI of Hyperpolarized Gas Enabled by Parahydrogen and Rh/TiO2 Heterogeneous Catalyst

    PubMed Central

    Barskiy, Danila A.; Coffey, Aaron M.; Truong, Milton L.; Salnikov, Oleg G.; Khudorozhkov, Alexander K.; Inozemtseva, Elizaveta A.; Prosvirin, Igor P.; Bukhtiyarov, Valery I.; Waddell, Kevin W.; Koptyug, Igor V.

    2015-01-01

    Several supported metal catalysts were synthesized, characterized, and tested in heterogeneous hydrogenation of propene with parahydrogen to maximize nuclear spin hyperpolarization of propane gas using parahydrogen induced polarization (PHIP). The Rh/TiO2 catalyst with a metal particle size of 1.6 nm was found to be the most active and effective in the pairwise hydrogen addition and robust, demonstrating reproducible results with multiple hydrogenation experiments and stability for ≥1.5 years. 3D 1H magnetic resonance imaging (MRI) of 1 % hyperpolarized flowing gas with microscale spatial resolution (625 × 625 × 625 μm3) and large imaging matrix (128 × 128 × 32) was demonstrated by using a preclinical 4.7 T scanner and 17.4 s imaging scan time. PMID:24961814

  19. Application of Double Spin-Echo Spiral Chemical Shift Imaging to Rapid Metabolic Mapping of Hyperpolarized [1-13C]-Pyruvate

    PubMed Central

    Josan, Sonal; Yen, Yi-Fen; Hurd, Ralph; Pfefferbaum, Adolf; Spielman, Daniel; Mayer, Dirk

    2011-01-01

    Undersampled spiral CSI (spCSI) using a free induction decay (FID) acquisition allows sub-second metabolic imaging of hyperpolarized 13C. Phase correction of the FID acquisition can be difficult, especially with contributions from aliased out-of-phase peaks. This work extends the spCSI sequence by incorporating double spin-echo radiofrequency (RF) pulses to eliminate the need for phase correction and obtain high quality spectra in magnitude mode. The sequence also provides an added benefit of attenuating signal from flowing spins, which can otherwise contaminate signal in the organ of interest. The refocusing pulses can potentially lead to a loss of hyperpolarized magnetization in dynamic imaging due to flow of spins through the fringe field of the RF coil, where the refocusing pulses fail to provide complete refocusing. Care must be taken for dynamic imaging to ensure that the spins remain within the B1-homogeneous sensitive volume of the RF coil. PMID:21316280

  20. Monitoring tumor response of prostate cancer to radiation therapy by multi-parametric 1H and hyperpolarized 13C magnetic resonance imaging

    NASA Astrophysics Data System (ADS)

    Zhang, Vickie Yi

    Radiation therapy is one of the most common curative therapies for patients with localized prostate cancer, but despite excellent success rates, a significant number of patients suffer post- treatment cancer recurrence. The accurate characterization of early tumor response remains a major challenge for the clinical management of these patients. Multi-parametric MRI/1H MR spectroscopy imaging (MRSI) has been shown to increase the diagnostic performance in evaluating the effectiveness of radiation therapy. 1H MRSI can detect altered metabolic profiles in cancerous tissue. In this project, the concentrations of prostate metabolites from snap-frozen biopsies of recurrent cancer after failed radiation therapy were correlated with histopathological findings to identify quantitative biomarkers that predict for residual aggressive versus indolent cancer. The total choline to creatine ratio was significantly higher in recurrent aggressive versus indolent cancer, suggesting that use of a higher threshold tCho/Cr ratio in future in vivo 1H MRSI studies could improve the selection and therapeutic planning for patients after failed radiation therapy. Varying radiation doses may cause a diverse effect on prostate cancer micro-environment and metabolism, which could hold the key to improving treatment protocols for individual patients. The recent development and clinical translation of hyperpolarized 13C MRI have provided the ability to monitor both changes in the tumor micro-environment and its metabolism using a multi-probe approach, [1-13C]pyruvate and 13C urea, combined with 1H Multi-parametric MRI. In this thesis, hyperpolarized 13C MRI, 1H dynamic contrast enhancement, and diffusion weighted imaging were used to identify early radiation dose response in a transgenic prostate cancer model. Hyperpolarized pyruvate to lactate metabolism significantly decreased in a dose dependent fashion by 1 day after radiation therapy, prior to any changes observed using 1H DCE and diffusion

  1. Direct and cost-efficient hyperpolarization of long-lived nuclear spin states on universal 15N2-diazirine molecular tags

    PubMed Central

    Theis, Thomas; Ortiz, Gerardo X.; Logan, Angus W. J.; Claytor, Kevin E.; Feng, Yesu; Huhn, William P.; Blum, Volker; Malcolmson, Steven J.; Chekmenev, Eduard Y.; Wang, Qiu; Warren, Warren S.

    2016-01-01

    Conventional magnetic resonance (MR) faces serious sensitivity limitations which can be overcome by hyperpolarization methods, but the most common method (dynamic nuclear polarization) is complex and expensive, and applications are limited by short spin lifetimes (typically seconds) of biologically relevant molecules. We use a recently developed method, SABRE-SHEATH, to directly hyperpolarize 15N2 magnetization and long-lived 15N2 singlet spin order, with signal decay time constants of 5.8 and 23 minutes, respectively. We find >10,000-fold enhancements generating detectable nuclear MR signals that last for over an hour. 15N2-diazirines represent a class of particularly promising and versatile molecular tags, and can be incorporated into a wide range of biomolecules without significantly altering molecular function. PMID:27051867

  2. SU-E-QI-11: Measurement of Renal Pyruvate-To-Lactate Exchange with Hyperpolarized 13C MRI

    SciTech Connect

    Adamson, E; Johnson, K; Fain, S; Gordon, J

    2014-06-15

    Purpose: Previous work [1] modeling the metabolic flux between hyperpolarized [1-13C]pyruvate and [1-13C]lactate in magnetic resonance spectroscopic imaging (MRSI) experiments failed to account for vascular signal artifacts. Here, we investigate a method to minimize the vascular signal and its impact on the fidelity of metabolic modeling. Methods: MRSI was simulated for renal metabolism in MATLAB both with and without bipolar gradients. The resulting data were fit to a two-site exchange model [1], and the effects of vascular partial volume artifacts on kinetic modeling were assessed. Bipolar gradients were then incorporated into a gradient echo sequence to validate the simulations experimentally. The degree of diffusion weighting (b = 32 s/mm{sup 2}) was determined empirically from 1H imaging of murine renal vascular signal. The method was then tested in vivo using MRSI with bipolar gradients following injection of hyperpolarized [1-{sup 13}C]pyruvate (∼80 mM at 20% polarization). Results: In simulations, vascular signal contaminated the renal metabolic signal at resolutions as high as 2 × 2 mm{sup 2} due to partial volume effects. The apparent exchange rate from pyruvate to lactate (k{sub p}) was underestimated in the presence of these artifacts due to contaminating pyruvate signal. Incorporation of bipolar gradients suppressed vascular signal and improved the accuracy of kp estimation. Experimentally, the in vivo results supported the ability of bipolar gradients to suppress vascular signal. The in vivo exchange rate increased, as predicted in simulations, from k{sub p} = 0.012 s-{sup 1} to k{sub p} = 0.020-{sup 1} after vascular signal suppression. Conclusion: We have demonstrated the limited accuracy of the two-site exchange model in the presence of vascular partial volume artifacts. The addition of bipolar gradients suppressed vascular signal and improved model accuracy in simulations. Bipolar gradients largely affected kp estimation in vivo. Currently

  3. Shift from depolarizing to hyperpolarizing glycine action in rat auditory neurones is due to age-dependent Cl− regulation

    PubMed Central

    Ehrlich, Ingrid; Löhrke, Stefan; Friauf, Eckhard

    1999-01-01

    The inhibitory neurotransmitter glycine can elicit depolarizing responses in immature neurones. We investigated the changes in glycine responses and their ionic mechanism in developing neurones of the rat lateral superior olive (LSO), an auditory brainstem nucleus involved in sound localization. Whole-cell and gramicidin perforated-patch recordings were performed from visually identified LSO neurones in brain slices and glycine was pressure applied for 3–100 ms to the soma. Glycine-evoked currents were reversibly blocked by strychnine. They were mostly monophasic, but biphasic responses occurred in ∼30% of P8-11 neurones in perforated-patch recordings. In whole-cell recordings from P2-11 neurones, the reversal potential of glycine-evoked currents (EGly) was determined by the transmembranous Cl− gradient and corresponded closely to the Nernst potential for Cl−, regardless of age. This indicates that Cl− is the principle ion permeating glycine receptors, but is also consistent with a low relative (10–20%) permeability for HCO3−. The Cl− gradient also determined the polarity and amplitude of glycine-evoked membrane potential changes. Leaving the native intracellular [Cl−] undisturbed with gramicidin perforated-patch recordings, we found a highly significant, age-dependent change of EGly from −46.8 ± 1.8 mV (P1-4, n = 28) to −67.6 ± 3.3 mV (P5-8, n = 10) to −82.2 ± 4.1 mV (P9–11, n = 18). The majority of P1–4 neurones were depolarized by glycine (∼80%) and spikes were evoked in ∼30%. In contrast, P9–11 neurones were hyperpolarized. In perforated-patch recordings, EGly was influenced by the voltage protocol and the glycine application interval; it could be shifted in the positive and negative direction. For a given application interval, these shifts were always larger in P1–4 than in P8–11 neurones, pointing to less effective Cl− regulation mechanisms in younger neurones. Furosemide (frusemide), a blocker of cation

  4. 2D multinuclear NMR, hyperpolarized xenon and gas storage in organosilica nanochannels with crystalline order in the walls.

    PubMed

    Comotti, Angiolina; Bracco, Silvia; Valsesia, Patrizia; Ferretti, Lisa; Sozzani, Piero

    2007-07-11

    The combination of 2D 1H-13C and 1H-29Si solid state NMR, hyperpolarized 129Xe NMR, synchrotron X-ray diffraction, together with adsorption measurements of vapors and gases for environmental and energetic relevance, was used to investigate the structure and the properties of periodic mesoporous hybrid p-phenylenesilica endowed with crystalline order in the walls. The interplay of 1H, 13C, and 29Si in the 2D heteronuclear correlation NMR measurements, together with the application of Lee-Goldburg homonuclear decoupling, revealed the spatial relationships (<5 angstroms) among various spin-active nuclei of the framework. Indeed, the through-space correlations in the 2D experiments evidenced, for the first time, the interfaces of the matrix walls with guest molecules confined in the nanochannels. Organic-inorganic and organic-organic heterogeneous interfaces between the matrix and the guests were identified. The open-pore structure and the easy accessibility of the nanochannels to the gas phase have been demonstrated by highly sensitive hyperpolarized (HP) xenon NMR, under extreme xenon dilution. Two-dimensional exchange experiments showed the exchange time to be as short as 2 ms. Through variable-temperature HP 129Xe NMR experiments we were able to achieve an unprecedented description of the nanochannel space and surface, a physisorption energy of 13.9 kJ mol-1, and the chemical shift value of xenon probing the internal surfaces. These results prompted us to measure the high storage capacity of the matrix towards benzene, hexafluorobenzene, ethanol, and carbon dioxide. Both host-guest, CH...pi, and OH...pi interactions contribute to the stabilization of the aromatic guests (benzene and hexafluorobenzene) on the extended surfaces. The full carbon dioxide loading in the channels could be detected by synchrotron radiation X-ray diffraction experiments. The selective adsorption of carbon dioxide (ca. 90 wt %) vs that of oxygen and hydrogen, together with the permanent

  5. Niflumic acid hyperpolarizes the smooth muscle cells by opening BK(Ca) channels through ryanodine-sensitive Ca(2+) release in spiral modiolar artery.

    PubMed

    Li, Li; Ma, Ke-Tao; Zhao, Lei; Si, Jun-Qiang

    2008-12-25

    The mechanism by which niflumic acid (NFA), a Cl(-) channel antagonist, hyperpolarizes the smooth muscle cells (SMCs) of cochlear spiral modiolar artery (SMA) was explored. Guinea pigs were used as subjects and perforated patch clamp and intracellular recording technique were used to observe NFA-induced response of SMC in the acutely isolated SMA preparation. The results showed that bath application of NFA, indanyloxyacetic acid 94 (IAA-94) and disodium 4,4'-diisothiocyanatostilbene-2,2'-disulfonate (DIDS) caused hyperpolarization and evoked outward currents in all cells at low resting potential (RP), but had no effects in cells at high RP. In the low RP SMCs, the average RP was about (-42.47+/-1.38) mV (n=24). Application of NFA (100 mumol/L), IAA-94 (10 mumol/L) and DIDS (200 mumol/L) shifted the RP to (13.7+/-4.3) mV (n=9, P<0.01), (11.4+/-4.2) mV (n=7, P<0.01) and (12.3+/-3.7) mV (n=8, P<0.01), respectively. These drug-induced responses were in a concentration-dependent manner. NFA-induced hyperpolarization and outward current were almost blocked by charybdotoxin (100 nmol/L), iberiotoxin (100 nmol/L), tetraethylammonium (10 mmol/L), BAPTA-AM (50 mumol/L), ryanodine (10 mumol/L) and caffeine (0.1-10 mmol/L), respectively, but not by nifedipine (100 mumol/L), CdCl2 (100 mumol/L) and Ca(2+)-free medium. It is concluded that NFA induces a release of intracellular calcium from the Ca(2+) stores and the released intracellular calcium in turn causes concentration-dependent and reversible hyperpolarization and evokes outward currents in the SMCs of the cochlear SMA via activation of the Ca(2+)-activated potassium channels.

  6. Detecting response of rat C6 glioma tumors to radiotherapy using hyperpolarized [1- 13C]pyruvate and 13C magnetic resonance spectroscopic imaging.

    PubMed

    Day, Sam E; Kettunen, Mikko I; Cherukuri, Murali Krishna; Mitchell, James B; Lizak, Martin J; Morris, H Douglas; Matsumoto, Shingo; Koretsky, Alan P; Brindle, Kevin M

    2011-02-01

    We show here that hyperpolarized [1-(13) C]pyruvate can be used to detect treatment response in a glioma tumor model; a tumor type where detection of response with (18) fluoro-2-deoxyglucose, using positron emission tomography, is limited by the high background signals from normal brain tissue. (13) C chemical shift images acquired following intravenous injection of hyperpolarized [1-(13) C]pyruvate into rats with implanted C6 gliomas showed significant labeling of lactate within the tumors but comparatively low levels in surrounding brain.Labeled pyruvate was observed at high levels in blood vessels above the brain and from other major vessels elsewhere but was detected at only low levels in tumor and brain.The ratio of hyperpolarized (13) C label in tumor lactate compared to the maximum pyruvate signal in the blood vessels was decreased from 0.38 ± 0.16 to 0.23 ± 0.13, (a reduction of 34%) by 72 h following whole brain irradiation with 15 Gy.

  7. Lys-[Leu8,des-Arg9]-bradykinin blocks lipopolysaccharide-induced SHR aorta hyperpolarization by inhibition of Ca(++)- and ATP-dependent K+ channels.

    PubMed

    Farias, Nelson C; Feres, Teresa; Paiva, Antonio C M; Paiva, Therezinha B

    2004-09-13

    The mediators involved in the hyperpolarizing effects of lipopolysaccharide and of the bradykinin B1 receptor agonist des-Arg9-bradykinin on the rat aorta were investigated by comparing the responses of aortic rings of spontaneously hypertensive and normotensive Wistar rats. Endothelized rings from hypertensive rats were hyperpolarized by des-Arg9-bradykinin and lipopolysaccharide, whereas de-endothelized rings responded to lipopolysaccharide but not to des-Arg9-bradykinin. In endothelized preparations, the responses to des-Arg9-bradykinin were inhibited by Nomega-nitro-L-arginine and iberiotoxin. De-endothelized ring responses to lipopolysaccharide were inhibited by iberiotoxin, glibenclamide and B1 antagonist Lys-[Leu8,des-Arg9]-bradykinin. This antagonist also inhibited hyperpolarization by des-Arg9-bradykinin and by the á2-adrenoceptor agonist, brimonidine. Our results indicate that Ca(2+)-sensitive K+ channels are the final mediators of the responses to des-Arg9-bradykinin, whereas both Ca(2+)- and ATP-sensitive K+ channels mediate the responses to lipopolysaccharide. The inhibitory effects of Lys-[Leu8,des-Arg9]-bradykinin is due to a direct action on Ca(2+)- and ATP-sensitive potassium channels.

  8. In vivo measurement of aldehyde dehydrogenase-2 activity in rat liver ethanol model using dynamic MRSI of hyperpolarized [1-(13) C]pyruvate.

    PubMed

    Josan, Sonal; Xu, Tao; Yen, Yi-Fen; Hurd, Ralph; Ferreira, Julio; Chen, Che-Hong; Mochly-Rosen, Daria; Pfefferbaum, Adolf; Mayer, Dirk; Spielman, Daniel

    2013-06-01

    To date, measurements of the activity of aldehyde dehydrogenase-2 (ALDH2), a critical mitochondrial enzyme for the elimination of certain cytotoxic aldehydes in the body and a promising target for drug development, have been largely limited to in vitro methods. Recent advancements in MRS of hyperpolarized (13) C-labeled substrates have provided a method to detect and image in vivo metabolic pathways with signal-to-noise ratio gains greater than 10 000-fold over conventional MRS techniques. However aldehydes, because of their toxicity and short T1 relaxation times, are generally poor targets for such (13) C-labeled studies. In this work, we show that dynamic MRSI of hyperpolarized [1-(13) C]pyruvate and its conversion to [1-(13) C]lactate can provide an indirect in vivo measurement of ALDH2 activity via the concentration of NADH (nicotinamide adenine dinucleotide, reduced form), a co-factor common to both the reduction of pyruvate to lactate and the oxidation of acetaldehyde to acetate. Results from a rat liver ethanol model (n = 9) show that changes in (13) C-lactate labeling following the bolus injection of hyperpolarized pyruvate are highly correlated with changes in ALDH2 activity (R(2) = 0.76).

  9. Enhancing the [13C]bicarbonate signal in cardiac hyperpolarized [1-13C]pyruvate MRS studies by infusion of glucose, insulin and potassium.

    PubMed

    Lauritzen, Mette Hauge; Laustsen, Christoffer; Butt, Sadia Asghar; Magnusson, Peter; Søgaard, Lise Vejby; Ardenkjær-Larsen, Jan Henrik; Åkeson, Per

    2013-11-01

    A change in myocardial metabolism is a known effect of several diseases. MRS with hyperpolarized (13)C-labelled pyruvate is a technique capable of detecting changes in myocardial pyruvate metabolism, and has proven to be useful for the evaluation of myocardial ischaemia in vivo. However, during fasting, the myocardial glucose oxidation is low and the fatty acid oxidation (β-oxidation) is high, which complicates the interpretation of pyruvate metabolism with the technique. The aim of this study was to investigate whether the infusion of glucose, insulin and potassium (GIK) could increase the myocardial glucose oxidation in the citric acid cycle, reflected as an increase in the [(13)C]bicarbonate signal in cardiac hyperpolarized [1-(13)C]pyruvate MRS measurements in fasted rats. Two groups of rats were infused with two different doses of GIK and investigated by MRS after injection of hyperpolarized [1-(13)C]pyruvate. No [(13)C]bicarbonate signal could be detected in the fasted state. However, a significant increase in the [(13)C]bicarbonate signal was observed by the infusion of a high dose of GIK. This study demonstrates that a high [(13)C]bicarbonate signal can be achieved by GIK infusion in fasted rats. The increased [(13)C]bicarbonate signal indicates an increased flux of pyruvate through the pyruvate dehydrogenase enzyme complex and an increase in myocardial glucose oxidation through the citric acid cycle.

  10. In vivo measurement of ethanol metabolism in the rat liver using magnetic resonance spectroscopy of hyperpolarized [1-13C]pyruvate

    PubMed Central

    Spielman, Daniel M.; Mayer, Dirk; Yen, Yi-Fen; Tropp, James; Hurd, Ralph E.; Pfefferbaum, Adolf

    2009-01-01

    [1-13C]pyruvate is readily polarizable substrate that has been the subject of numerous magnetic resonance spectroscopy (MRS) studies of in vivo metabolism. In this work, 13C-MRS of hyperpolarized [1-13C]pyruvate is used to interrogate a metabolic pathway involved in neither aerobic nor anaerobic metabolism. In particular, ethanol consumption leads to altered liver metabolism, which when excessive is associated with adverse medical conditions including fatty liver disease, hepatitis, cirrhosis, and cancer. Here we present a method for noninvasively monitoring this important process in vivo. Following the bolus injection of hyperpolarized [1-13C]pyruvate, we demonstrate a significantly increased rat liver lactate production rate with the co-administration of ethanol (P = 0.0016 unpaired t-test). The affect is attributable to increased liver nicotinamide adenine dinucleotide (NADH) associated with ethanol metabolism in combination with NADH's role as a coenzyme in pyruvate to lactate conversion. Beyond studies of liver metabolism, this novel in vivo assay of changes in NADH levels makes hyperpolarized [1-13C]pyruvate a potentially viable substrate for studying the multiple in vivo metabolic pathways that use NADH (or NAD+) as a coenzyme, thus broadening the range of applications that have been discussed in the literature to date. PMID:19526498

  11. Saturation-recovery metabolic-exchange rate imaging with hyperpolarized [1-13C] pyruvate using spectral-spatial excitation.

    PubMed

    Schulte, Rolf F; Sperl, Jonathan I; Weidl, Eliane; Menzel, Marion I; Janich, Martin A; Khegai, Oleksandr; Durst, Markus; Ardenkjaer-Larsen, Jan Henrik; Glaser, Steffen J; Haase, Axel; Schwaiger, Markus; Wiesinger, Florian

    2013-05-01

    Within the last decade hyperpolarized [1-13C] pyruvate chemical-shift imaging has demonstrated impressive potential for metabolic MR imaging for a wide range of applications in oncology, cardiology, and neurology. In this work, a highly efficient pulse sequence is described for time-resolved, multislice chemical shift imaging of the injected substrate and obtained downstream metabolites. Using spectral-spatial excitation in combination with single-shot spiral data acquisition, the overall encoding is evenly distributed between excitation and signal reception, allowing the encoding of one full two-dimensional metabolite image per excitation. The signal-to-noise ratio can be flexibly adjusted and optimized using lower flip angles for the pyruvate substrate and larger ones for the downstream metabolites. Selectively adjusting the excitation of the down-stream metabolites to 90° leads to a so-called "saturation-recovery" scheme with the detected signal content being determined by forward conversion of the available pyruvate. In case of repetitive excitations, the polarization is preserved using smaller flip angles for pyruvate. Metabolic exchange rates are determined spatially resolved from the metabolite images using a simplified two-site exchange model. This novel contrast is an important step toward more quantitative metabolic imaging. Goal of this work was to derive, analyze, and implement this "saturation-recovery metabolic exchange rate imaging" and demonstrate its capabilities in four rats bearing subcutaneous tumors.

  12. Control of the hyperpolarization-activated cation current by external anions in rabbit sino-atrial node cells.

    PubMed Central

    Frace, A M; Maruoka, F; Noma, A

    1992-01-01

    1. Effects of varying concentrations of anions on the hyperpolarization-activated current (I(f)) were studied in myocytes isolated from the rabbit sino-atrial node. Substituting Cs+ for the intracellular K+ clearly separated I(f) from the delayed rectifier K+ current. Control properties, including gating kinetics and ion selectivity, similar to previous studies were obtained. 2. Substitution of extracellular Cl- with larger anions including isethionate, glutamate, acetate, and aspartate, reduced the amplitude of I(f) without changing the reversal potential. Substitution with small anions such as iodide or nitrate supported an intact I(f). These effects were reproduced in the excised outside-out patch conformation. 3. The conductance for I(f) was a saturating function of the extracellular Cl- concentration ([Cl-]o) with an equilibrium binding constant (K1/2) of 11 mM and a slope factor of about 1 when substituted with large anions. Total removal of small anions completely abolished I(f). 4. The voltage-dependent gating of I(f) was not affected by changing ([Cl-]o), suggesting that Cl- modulates conductance properties of I(f). 5. The results indicate that I(f) conductance is unique in that it is dependent on an extracellular anion (Cl-), yet it is carried exclusively by cations, K+ and Na+. These effects are independent of any measurable voltage-dependent gating parameters. PMID:1281504

  13. A rapid method for direct detection of metabolic conversion and magnetization exchange with application to hyperpolarized substrates

    NASA Astrophysics Data System (ADS)

    Larson, Peder E. Z.; Kerr, Adam B.; Leon Swisher, Christine; Pauly, John M.; Vigneron, Daniel B.

    2012-12-01

    In this work, we present a new MR spectroscopy approach for directly observing nuclear spins that undergo exchange, metabolic conversion, or, generally, any frequency shift during a mixing time. Unlike conventional approaches to observe these processes, such as exchange spectroscopy (EXSY), this rapid approach requires only a single encoding step and thus is readily applicable to hyperpolarized MR in which the magnetization is not replenished after T1 decay and RF excitations. This method is based on stimulated-echoes and uses phase-sensitive detection in conjunction with precisely chosen echo times in order to separate spins generated during the mixing time from those present prior to mixing. We are calling the method Metabolic Activity Decomposition Stimulated-echo Acquisition Mode or MAD-STEAM. We have validated this approach as well as applied it in vivo to normal mice and a transgenic prostate cancer mouse model for observing pyruvate-lactate conversion, which has been shown to be elevated in numerous tumor types. In this application, it provides an improved measure of cellular metabolism by separating [1-13C]-lactate produced in tissue by metabolic conversion from [1-13C]-lactate that has flowed into the tissue or is in the blood. Generally, MAD-STEAM can be applied to any system in which spins undergo a frequency shift.

  14. Regional Mapping of Gas Uptake by Blood and Tissue in the Human Lung using Hyperpolarized Xenon-129 MRI

    PubMed Central

    Qing, Kun; Ruppert, Kai; Jiang, Yun; Mata, Jaime F.; Miller, G. Wilson; Shim, Y. Michael; Wang, Chengbo; Ruset, Iulian C.; Hersman, F. William; Altes, Talissa A.; Mugler, John P.

    2013-01-01

    Purpose To develop a breath-hold acquisition for regional mapping of ventilation and the fractions of hyperpolarized xenon-129 (Xe129) dissolved in tissue (lung parenchyma and plasma) and red blood cells (RBCs), and to perform an exploratory study to characterize data obtained in human subjects. Materials and Methods A three-dimensional, multi-echo, radial-trajectory pulse sequence was developed to obtain ventilation (gaseous Xe129), tissue and RBC images in healthy subjects, smokers and asthmatics. Signal ratios (total dissolved Xe129 to gas, tissue-to-gas, RBC-to-gas and RBC-to-tissue) were calculated from the images for quantitative comparison. Results Healthy subjects demonstrated generally uniform values within coronal slices, and a gradient in values along the anterior-to-posterior direction. In contrast, images and associated ratio maps in smokers and asthmatics were generally heterogeneous and exhibited values mostly lower than those in healthy subjects. Whole-lung values of total dissolved Xe129 to gas, tissue-to-gas, and RBC-to-gas ratios in healthy subjects were significantly larger than those in diseased subjects. Conclusion Regional maps of tissue and RBC fractions of dissolved Xe129 were obtained from a short breath-hold acquisition, well tolerated by healthy volunteers and subjects with obstructive lung disease. Marked differences were observed in spatial distributions and overall amounts of Xe129 dissolved in tissue and RBCs among healthy subjects, smokers and asthmatics. PMID:23681559

  15. Differences in Vascular Nitric Oxide and Endothelium-Derived Hyperpolarizing Factor Bioavailability in African Americans and Whites

    PubMed Central

    Ozkor, Muhiddin A; Rahman, Ayaz M; Murrow, Jonathan R; Kavtaradze, Nino; Lin, Ji; Manatunga, Amita; Hayek, Salim; Quyyumi, Arshed A

    2014-01-01

    Objectives Abnormalities in nitric oxide (NO) bioavailability have been reported in African Americans. Whether there are differences in endothelium-derived hyperpolarizing factor (EDHF) in addition to NO between African Americans and whites, and how these affect physiologic vasodilation remains unknown. We hypothesized that the bioavailability of vascular NO and EDHF, at rest and with pharmacologic and physiologic vasodilation, varies between white and African Americans. Approach and Results In 74 white and 86 African American subjects without known cardiovascular disease risk factors, forearm blood flow (FBF) was measured using plethysmography at rest and during inhibition of NO with NG-monomethyl-L-arginine (L-NMMA) and/or of K+Ca channels (EDHF) with tetraethylammonium (TEA). The reduction in resting FBF was greater with L-NMMA (p=0.019) and similar with TEA in whites compared to African Americans. Vasodilation with bradykinin, acetylcholine, and sodium nitroprusside was lower in African Americans compared to whites (all p<0.0001). Inhibition with L-NMMA was greater in whites compared to African Americans with bradykinin, acetylcholine, and exercise. Inhibition with TEA was lower in African Americans with bradykinin, but greater during exercise and with acetylcholine. Conclusions The contribution to both resting and stimulus-mediated vasodilator tone of NO is greater in whites compared to African Americans. EDHF partly compensates for the reduced NO release in exercise and acetylcholine-mediated vasodilation in African Americans. Preserved EDHF but reduced NO bioavailability and sensitivity characterizes the vasculature in healthy African Americans. PMID:24675657

  16. ATP-dependent acidification and tonoplast hyperpolarization in isolated vacuoles from green suspension cells of Chenopodium rubrum L

    PubMed Central

    Bentrup, F.-W.; Gogarten-Boekels, M.; Hoffmann, B.; Gogarten, J. P.; Baumann, C.

    1986-01-01

    The tonoplast of isolated vacuoles from photoautotrophic suspension cells of Chenopodium rubrum L. was studied by means of the patch-clamp technique. In a symmetrical K+ concentration of 46 mM, similar to in vivo conditions, the tonoplast displayed a membrane potential near zero and a linear current-voltage relationship with a mean slope of 1.0 S/m2. ATP at 2 mM hyperpolarized the tonoplast (vacuole positive) by 15-20 mV and, in a parallel experiment, acidified the vacuole (outside pH 7.0) to pH 5.0, as monitored by accumulation of acridine orange. Analysis of the voltage-clamp current indicates a 2-fold, ATP-dependent increase of the membrane capacitance, from 4 to 8 mF/m2, and an ATP-independent, unidentified ion channel having a mean opening time of about 5 msec and a conductivity of 0.5-1.0 pS. PMID:16593685

  17. Role of metabolic modulator Bet-CA in altering mitochondrial hyperpolarization to suppress cancer associated angiogenesis and metastasis

    PubMed Central

    Saha, Suchandrima; Ghosh, Monisankar; Dutta, Samir Kumar

    2016-01-01

    Solid tumors characteristically reflect a metabolic switching from glucose oxidation to glycolysis that plays a fundamental role in angiogenesis and metastasis to facilitate aggressive tumor outcomes. Hyperpolarized mitochondrial membrane potential is a manifestation of malignant cells that compromise the intrinsic pathways of apoptosis and confer a suitable niche to promote the cancer associated hallmark traits. We have previously reported that co-drug Bet-CA selectively targets cancer cells by inducing metabolic catastrophe without a manifest in toxicity. Here we report that the same molecule at a relatively lower concentration deregulates the cardinal phenotypes associated with angiogenesis and metastasis. In mice syngeneic 4T1 breast cancer model, Bet-CA exhibited effective abrogation of angiogenesis and concomitantly obliterated lung metastasis consistent with altered mitochondrial bioenergetics. Furthermore, Bet-CA significantly lowered vascular endothelial growth factor (VEGF) levels and obviated matrix metalloproteases (MMP-2/9) production directly to the criterion where abrogation of autocrine VEGF/VEGFR2 signalling loop was documented. In vitro studies anticipatedly documented the role of Bet-CA in inhibiting actin remodeling, lamellipodia formation and cell membrane ruffling to constitutively suppress cell motility and invasion. Results comprehensively postulate that Bet-CA, a mitochondria targeting metabolic modulator may serve as an excellent candidate for combating angiogenesis and metastasis. PMID:27003027

  18. Enhancement in low field nuclear magnetic resonance with a high-Tc superconducting quantum interference device and hyperpolarized 3He

    NASA Astrophysics Data System (ADS)

    Liao, Shu-Hsien; Yang, Hong-Chang; Horng, Herng-Er; Chen, Hsin-Hsien; Yang, Shieh-Yueh; Chen, Ming-Jye; Yang, Chang-Hau

    2008-09-01

    In this work, we present a design that improves signals produced by nuclear magnetic resonance (NMR) and magnetic resonance imaging by using optical pumping and a high-Tc superconducting quantum interference device (SQUID) magnetometer. In our design for a NMR detection system, a pickup coil is coupled to the spin procession of a H3e nucleus; the input coil is coupled to a high-Tc SQUID magnetometer; and the capacitor is connected in series to form a tank circuit resonating at the Larmor frequency of the H3e nucleus in the measuring field. A signal-to-noise ratio gain of 2.67 over a conventional Faraday detection coil was obtained with the high-Tc SQUID detection system in a measuring magnetic field equaling 0.1128 mT, at which the central frequency was 3.66 kHz for H3e nucleus. The improvement in the NMR signal for large-size, hyperpolarized H3e coupled to a high-Tc SQUID-based spectrometer in low magnetic fields at room temperature is significant compared to that without flux coupling. This result may be of interest given its potential for use in a low field imager.

  19. Effect of stimulation and hyperpolarization on non-electrolyte and sodium permeability in perfused axons of squid.

    PubMed

    Hidalgo, C; Latorre, R

    1970-11-01

    1. The permeability for micro-injected [(3)H]ethylene glycol was measured in resting state and during stimulation at 100/sec in squid giant axons. No detectable changes during electrical activity were observed.2. The influxes of urethane, tritiated water, ethylene glycol, urea and sodium were measured in internally perfused squid axons. Ethylene glycol and urea influxes were determined simultaneously with sodium influxes. The electrical stimulation of the fibre produced an increase in the influx of sodium but did not alter the influxes of the non-electrolytes listed above.3. Experiments were done with the combined voltage clamp-perfusion technique. The influxes of ethylene glycol and sodium were simultaneously measured in resting state and during maximum sodium current under stimulation at 10/sec. The influx of sodium increased in these conditions but the influx of ethylene glycol remained constant. In some experiments, the fibre was hyperpolarized to 10 or 20 mV, above the resting potential and the influxes of ethylene glycol and sodium were measured. The sodium influx decreased to 60% at 20 mV above the resting potential whereas the influx of ethylene glycol remained constant.4. These results indicate that in the giant axons of the squid Dosidicus gigas, sodium and non-electrolytes fluxes are not coupled.

  20. Ascorbate elevates perfusion pressure in the bovine extraocular long posterior ciliary artery: role of endothelium-derived hyperpolarizing factor (EDHF).

    PubMed

    Stirrat, Alison; Nelli, Silvia; McGuckin, Alicia; Ho, Vivian Wing Man; Wilson, William S; Martin, William

    2006-03-18

    Ascorbate blocks agonist-induced, endothelium-derived hyperpolarizing factor (EDHF)-mediated vasodilatation in the bovine perfused ciliary artery and this is associated with a rise in perfusion pressure. We now report the origins of this ascorbate-induced rise in perfusion pressure. In segments of ciliary artery perfused at 2.5 ml/min, the addition of ascorbate (10-150 microM) enhanced U46619-induced perfusion pressure. Ascorbate produced no enhancement in the absence of U46619, suggesting that its effects resulted not from a constrictor action but through removal of a tonic vasodilator influence. Experiments revealed the endothelial source of this vasodilator influence, and EDHF, but not nitric oxide or prostanoids, appeared to be involved. The ascorbate-induced enhancement of vasoconstrictor tone was not seen in a static myograph or in segments perfused at low rates of flow, but was seen at flow rates of 2.5 ml(-1) and above. We conclude that ascorbate augments vasoconstrictor tone through inhibition of flow-induced EDHF activity.

  1. Long-range diffusion of hyperpolarized 3He in explanted normal and emphysematous human lungs via magnetization tagging

    PubMed Central

    Woods, Jason C.; Yablonskiy, Dmitriy A.; Choong, Cliff K.; Chino, Kimiaki; Pierce, John A.; Hogg, James C.; Bentley, John; Cooper, Joel D.; Conradi, Mark S.; Macklem, Peter T.

    2007-01-01

    Long-range diffusivity of hyperpolarized 3He gas was measured from the decay rate of sinusoidally modulated longitudinal nuclear magnetization in three normal donor and nine severely emphysematous explanted human lungs. This (long-range) diffusivity, which we call Dsec, is measured over seconds and centimeters and is ~10 times smaller in healthy lungs (0.022 cm2/s) than the more traditionally measured Dmsec, which is measured over milliseconds and submillimeters. The increased restriction of Dsec reflects the complex, tortuous paths required to navigate long distances through the maze of branching peripheral airways. In emphysematous lungs, Dsec is substantially increased, with some regions showing nearly the unrestricted value of the self-diffusion coefficient (0.88 cm2/s for dilute 3He in air, a 40-fold increase). This suggests the presence of large collateral pathways opened by alveolar destruction that bypass the airways proper. This destruction was confirmed by comparison with histology in seven lungs and by removal of trapped gas via holes in the pleural surface in five lungs. PMID:16024528

  2. Diffusion of hyperpolarized 129Xe in the lung: a simplified model of 129Xe septal uptake and experimental results

    NASA Astrophysics Data System (ADS)

    Patz, Samuel; Muradyan, Iga; Hrovat, Mirko I.; Dabaghyan, Mikayel; Washko, George R.; Hatabu, Hiroto; Butler, James P.

    2011-01-01

    We used hyperpolarized 129Xe NMR to measure pulmonary alveolar surface area per unit gas volume SA/Vgas, alveolar septal thickness h and capillary transit time τ, three critical determinants of the lung's primary role as a gas exchange organ. An analytical solution for a simplified diffusion model is described, together with a modification of the xenon transfer contrast imaging technique utilizing 90° radio-frequency pulses applied to the dissolved phase, rather than traditional 180° pulses. With this approach, three-dimensional (3D) maps of SA/Vgas were obtained. We measured global SA/Vgas, h and τ in four normal subjects, two subjects with mild interstitial lung disease (ILD) and two subjects with mild chronic obstructive pulmonary disease (COPD). In normals, SA/Vgas decreased with increasing lung volume from ~320 to 80 cm-1 both h~13 μm and τ~1.5 s were relatively constant. For the two ILD subjects, h was, respectively, 36 and 97% larger than normal, quantifying an increased gas/blood tissue barrier; SA/Vgas and τ were normal. The two COPD subjects had SA/Vgas values ~25% that of normals, quantifying septal surface loss in emphysema; h and τ were normal. These are the first noninvasive, non-radiation-based, quantitative measurements of h and τ in patients with pulmonary disease.

  3. ZD7288, a selective hyperpolarization-activated cyclic nucleotide-gated channel blocker, inhibits hippocampal synaptic plasticity

    PubMed Central

    Zhang, Xiao-xue; Min, Xiao-chun; Xu, Xu-lin; Zheng, Min; Guo, Lian-jun

    2016-01-01

    The selective hyperpolarization-activated cyclic nucleotide-gated (HCN) channel blocker 4-(N-ethyl-N-phenylamino)-1,2-dimethyl-6-(methylamino) pyrimidinium chloride (ZD7288) blocks the induction of long-term potentiation in the perforant path–CA3 region in rat hippocampus in vivo. To explore the mechanisms underlying the action of ZD7288, we recorded excitatory postsynaptic potentials in perforant path–CA3 synapses in male Sprague-Dawley rats. We measured glutamate content in the hippocampus and in cultured hippocampal neurons using high performance liquid chromatography, and determined intracellular Ca2+ concentration [Ca2+]i) using Fura-2. ZD7288 inhibited the induction and maintenance of long-term potentiation, and these effects were mirrored by the nonspecific HCN channel blocker cesium. ZD7288 also decreased glutamate release in hippocampal tissue and in cultured hippocampal neurons. Furthermore, ZD7288 attenuated glutamate-induced rises in [Ca2+]i in a concentration-dependent manner and reversed 8-Br-cAMP-mediated facilitation of these glutamate-induced [Ca2+]i rises. Our results suggest that ZD7288 inhibits hippocampal synaptic plasticity both glutamate release and resultant [Ca2+]i increases in rat hippocampal neurons. PMID:27335562

  4. Mechanisms of seizure-induced ‘transcriptional channelopathy’ of hyperpolarization-activated cyclic nucleotide gated (HCN) channels

    PubMed Central

    Richichi, Cristina; Brewster, Amy L.; Bender, Roland A.; Simeone, Timothy A.; Zha, Qinqin; Yin, Hong Z.; Weiss, John H.; Baram, Tallie Z.

    2008-01-01

    Epilepsy may result from abnormal function of ion channels, such as those caused by genetic mutations. Recently, pathological alterations of the expression or localization of normal channels have been implicated in epilepsy generation, and termed ‘acquired channelopathies’. Altered expression levels of the HCN channels--that conduct the hyperpolarization-activated current, Ih--have been demonstrated in hippocampus of patients with severe temporal lobe epilepsy as well as in animal models of temporal lobe and absence epilepsies. Here we probe the mechanisms for the altered expression of HCN channels which is provoked by seizures. In organotypic hippocampal slice cultures, seizure-like events selectively reduced HCN type 1 channel expression and increased HCN2 mRNA levels, as occurs in vivo. The mechanisms for HCN1 reduction involved Ca2+-permeable AMPA receptors-mediated Ca2+ influx, and subsequent activation of Ca2+/calmodulin-dependent protein kinase II. In contrast, upregulation of HCN2 expression was independent of these processes. The data demonstrate an orchestrated program for seizure-evoked transcriptional channelopathy involving the HCN channels, that may contribute to certain epilepsies. PMID:17964174

  5. Delivering strong (1)H nuclear hyperpolarization levels and long magnetic lifetimes through signal amplification by reversible exchange.

    PubMed

    Rayner, Peter J; Burns, Michael J; Olaru, Alexandra M; Norcott, Philip; Fekete, Marianna; Green, Gary G R; Highton, Louise A R; Mewis, Ryan E; Duckett, Simon B

    2017-04-04

    Hyperpolarization turns typically weak NMR and MRI responses into strong signals so that ordinarily impractical measurements become possible. The potential to revolutionize analytical NMR and clinical diagnosis through this approach reflect this area's most compelling outcomes. Methods to optimize the low-cost parahydrogen-based approach signal amplification by reversible exchange with studies on a series of biologically relevant nicotinamides and methyl nicotinates are detailed. These procedures involve specific (2)H labeling in both the agent and catalyst and achieve polarization lifetimes of ca 2 min with 50% polarization in the case of methyl-4,6-d2 -nicotinate. Because a 1.5-T hospital scanner has an effective (1)H polarization level of just 0.0005% this strategy should result in compressed detection times for chemically discerning measurements that probe disease. To demonstrate this technique's generality, we exemplify further studies on a range of pyridazine, pyrimidine, pyrazine, and isonicotinamide analogs that feature as building blocks in biochemistry and many disease-treating drugs.

  6. Synchronization modulation of Na/K pump molecules can hyperpolarize the membrane resting potential in intact fibers.

    PubMed

    Chen, Wei; Dando, Robin

    2007-02-01

    Previously, we have theoretically studied the possibility of electrical rhythmic entrainment of carrier-mediated ion transporters, and experimentally realized synchronization and acceleration of the Na/K pumping rate in the cell membrane of skeletal muscle fibers by a specially designed synchronization modulation electric field. In these studies we either used cut fibers under a voltage clamp or intact fibers, but in the presence of ion channels blockers. A question remained as to whether the field-induced activation observed in the pump molecules could effectively increase the intracellular ionic concentration and the membrane potential at physiological conditions. In this paper, we studied the effects of the field on intact fibers without any channel blockers. We monitored the field-induced changes in the ionic concentration gradient across the cell membrane and the membrane potential non-invasively by using a fluorescent probe and confocal microscopic imaging techniques. The results clearly show that the entrainment of the pump molecules by the synchronization modulation electric field can effectively increase the ionic concentration gradient, and hence, hyperpolarize the membrane potential.

  7. Band-selective chemical exchange saturation transfer imaging with hyperpolarized xenon-based molecular sensors

    NASA Astrophysics Data System (ADS)

    Meldrum, Tyler; Bajaj, Vikram S.; Wemmer, David E.; Pines, Alexander

    2011-12-01

    Molecular imaging based on saturation transfer in exchanging systems is a tool for amplified and chemically specific magnetic resonance imaging. Xenon-based molecular sensors are a promising category of molecular imaging agents in which chemical exchange of dissolved xenon between its bulk and agent-bound phases has been use to achieve sub-picomolar detection sensitivity. Control over the saturation transfer dynamics, particularly when multiple exchanging resonances are present in the spectra, requires saturation fields of limited bandwidth and is generally accomplished by continuous wave irradiation. We demonstrate instead how band-selective saturation sequences based on multiple pulse inversion elements can yield saturation bandwidth tuneable over a wide range, while depositing less RF power in the sample. We show how these sequences can be used in imaging experiments that require spatial-spectral and multispectral saturation. The results should be applicable to all CEST experiments and, in particular, will provide the spectroscopic control required for applications of arrays of xenon chemical sensors in microfluidic chemical analysis devices.

  8. Band-selective chemical exchange saturation transfer imaging with hyperpolarized xenon-based molecular sensors.

    PubMed

    Meldrum, Tyler; Bajaj, Vikram S; Wemmer, David E; Pines, Alexander

    2011-12-01

    Molecular imaging based on saturation transfer in exchanging systems is a tool for amplified and chemically specific magnetic resonance imaging. Xenon-based molecular sensors are a promising category of molecular imaging agents in which chemical exchange of dissolved xenon between its bulk and agent-bound phases has been use to achieve sub-picomolar detection sensitivity. Control over the saturation transfer dynamics, particularly when multiple exchanging resonances are present in the spectra, requires saturation fields of limited bandwidth and is generally accomplished by continuous wave irradiation. We demonstrate instead how band-selective saturation sequences based on multiple pulse inversion elements can yield saturation bandwidth tuneable over a wide range, while depositing less RF power in the sample. We show how these sequences can be used in imaging experiments that require spatial-spectral and multispectral saturation. The results should be applicable to all CEST experiments and, in particular, will provide the spectroscopic control required for applications of arrays of xenon chemical sensors in microfluidic chemical analysis devices.

  9. 5-Hydroxytryptamine1A receptor-activation hyperpolarizes pyramidal cells and suppresses hippocampal gamma oscillations via Kir3 channel activation.

    PubMed

    Johnston, April; McBain, Chris J; Fisahn, André

    2014-10-01

    Rhythmic cortical neuronal oscillations in the gamma frequency band (30-80 Hz, gamma oscillations) have been associated with cognitive processes such as sensory perception and integration, attention, learning, and memory. Gamma oscillations are disrupted in disorders for which cognitive deficits are hallmark symptoms such as schizophrenia and Alzheimer's disease.In vitro, various neurotransmitters have been found to modulate gamma oscillations. Serotonin(5-HT) has long been known to be important for both behavioural and cognitive functions such as learning and memory. Multiple 5-HT receptor subtypes are expressed in the CA3 region of the hippocampus and high doses of 5-HT reduce the power of induced gamma oscillations.Hypothesizing that 5-HT may have cell- and receptor subtype-specific modulatory effects, we investigated the receptor subtypes, cell types and cellular mechanisms engaged by 5-HT in the modulation of gamma oscillations in mice and rats. We found that 5-HT decreases the power of kainate-induced hippocampal gamma oscillations in both species via the 5-HT1A receptor subtype. Whole-cell patch clamp recordings demonstrated that this decrease was caused by a hyperpolarization of CA3 pyramidal cells and a reduction of their firing frequency, but not by alteration of inhibitory neurotransmission. Finally, our results show that the effect on pyramidal cells is mediated via the G protein-coupled receptor inwardly rectifying potassium channel Kir3.Our findings suggest this novel cellular mechanism as a potential target for therapies that are aimed at alleviating cognitive decline by helping the brain to maintain or re-establish normal gamma oscillation levels in neuropsychiatric and neurodegenerative disorders.

  10. Modulation of the hyperpolarization-activated current (Ih) by cyclic nucleotides in guinea-pig primary afferent neurons.

    PubMed Central

    Ingram, S L; Williams, J T

    1996-01-01

    1. Whole-cell patch-clamp recordings were made from dissociated guinea-pig nodose and trigeminal ganglion neurons in culture to study second messenger mechanisms of the hyperpolarization-activated current (Ih) modulation. 2. Prostaglandin E2 (PGE2) and forskolin modulate Ih in primary afferents by shifting the activation curve in the depolarizing direction and increasing the maximum amplitude. 3. The cAMP analogues, RP-cAMP-S (an inhibitor of protein kinase A (PKA)) and SP-cAMP-S (an activator of PKA), both shifted the activation curve of Ih to more depolarized potentials and occluded the effects of forskolin. These results suggest that Ih is modulated by a direct action of the cAMP analogues. 4. Superfusion of other cyclic nucleotide analogues (8-Br-cAMP, 8-(4-chlorophenylthio)-cAMP and 8-Br-cGMP) mimicked the actions of forskolin and PGE2, but dibutyryl cGMP, 5'-AMP and adenosine had no effect on Ih. 8-Br-cAMP and 8-Br-cGMP had similar concentration response profiles, suggesting that Ih has little nucleotide selectivity. 5. The inhibitor peptide (PKI), the catalytic subunit of PKA (C subunit) and phosphatase inhibitors (microcystin and okadaic acid) had no effect on forskolin modulation of Ih. 6. These results indicate that Ih is regulated by cyclic nucleotides in sensory neurons. Positive regulation of Ih by prostaglandins produced during inflammation may lead to depolarization and facilitation of repetitive activity, and thus contribute to sensitization to painful stimuli. PMID:8730586

  11. A robust protocol for regional evaluation of methacholine challenge in mouse models of allergic asthma using hyperpolarized 3He MRI.

    PubMed

    Thomas, Abraham C; Potts, Erin N; Chen, Ben T; Slipetz, Deborah M; Foster, W Michael; Driehuys, Bastiaan

    2009-06-01

    Hyperpolarized (HP) (3)He magnetic resonance imaging has been recently used to produce high-resolution images of pulmonary ventilation after methacholine (MCh) challenge in mouse models of allergic inflammation. This capability presents an opportunity to gain new insights about these models and to more sensitively evaluate new drug treatments in the pre-clinical setting. In the current study, we present our initial experience using two-dimensional (2D), time-resolved (3)He MRI of MCh challenge-induced airways hyperreactivity (AHR) to compare ovalbumin-sensitized and challenged (N = 8) mice to controls (N = 8). Imaging demonstrated that ovalbumin-sensitized and challenged animals exhibited many large ventilation defects even prior to MCh challenge (four out of eight) compared to no defects in the control animals. Additionally, the ovalbumin-sensitized and challenged animals experienced a greater number of ventilation defects (4.5 +/- 0.4) following MCh infusion than did controls (3.3 +/- 0.6). However, due to variability in MCh delivery that was specific to the small animal MRI environment, the difference in mean defect number was not statistically significant. These findings are reviewed in detail and a comprehensive solution to the variability problem is presented that has greatly enhanced the magnitude and reproducibility of the MCh response. This has permitted us to develop a new imaging protocol consisting of a baseline 3D image, a time-resolved 2D series during MCh challenge, and a post-MCh 3D image that reveals persistent ventilation defects.

  12. Increasing Pyruvate Dehydrogenase Flux as a Treatment for Diabetic Cardiomyopathy: A Combined 13C Hyperpolarized Magnetic Resonance and Echocardiography Study.

    PubMed

    Le Page, Lydia M; Rider, Oliver J; Lewis, Andrew J; Ball, Vicky; Clarke, Kieran; Johansson, Edvin; Carr, Carolyn A; Heather, Lisa C; Tyler, Damian J

    2015-08-01

    Although diabetic cardiomyopathy is widely recognized, there are no specific treatments available. Altered myocardial substrate selection has emerged as a candidate mechanism behind the development of cardiac dysfunction in diabetes. As pyruvate dehydrogenase (PDH) activity appears central to the balance of substrate use, we aimed to investigate the relationship between PDH flux and myocardial function in a rodent model of type 2 diabetes and to explore whether or not increasing PDH flux, with dichloroacetate, would restore the balance of substrate use and improve cardiac function. All animals underwent in vivo hyperpolarized [1-(13)C]pyruvate magnetic resonance spectroscopy and echocardiography to assess cardiac PDH flux and function, respectively. Diabetic animals showed significantly higher blood glucose levels (10.8 ± 0.7 vs. 8.4 ± 0.5 mmol/L), lower PDH flux (0.005 ± 0.001 vs. 0.017 ± 0.002 s(-1)), and significantly impaired diastolic function (transmitral early diastolic peak velocity/early diastolic myocardial velocity ratio [E/E'] 12.2 ± 0.8 vs. 20 ± 2), which are in keeping with early diabetic cardiomyopathy. Twenty-eight days of treatment with dichloroacetate restored PDH flux to normal levels (0.018 ± 0.002 s(-1)), reversed diastolic dysfunction (E/E' 14 ± 1), and normalized blood glucose levels (7.5 ± 0.7 mmol/L). The treatment of diabetes with dichloroacetate therefore restored the balance of myocardial substrate selection, reversed diastolic dysfunction, and normalized blood glucose levels. This suggests that PDH modulation could be a novel therapy for the treatment and/or prevention of diabetic cardiomyopathy.

  13. Near-unity nuclear polarization with an open-source 129Xe hyperpolarizer for NMR and MRI

    PubMed Central

    Nikolaou, Panayiotis; Coffey, Aaron M.; Walkup, Laura L.; Gust, Brogan M.; Whiting, Nicholas; Newton, Hayley; Barcus, Scott; Muradyan, Iga; Dabaghyan, Mikayel; Moroz, Gregory D.; Rosen, Matthew S.; Patz, Samuel; Barlow, Michael J.; Chekmenev, Eduard Y.; Goodson, Boyd M.

    2013-01-01

    The exquisite NMR spectral sensitivity and negligible reactivity of hyperpolarized xenon-129 (HP129Xe) make it attractive for a number of magnetic resonance applications; moreover, HP129Xe embodies an alternative to rare and nonrenewable 3He. However, the ability to reliably and inexpensively produce large quantities of HP129Xe with sufficiently high 129Xe nuclear spin polarization (PXe) remains a significant challenge—particularly at high Xe densities. We present results from our “open-source” large-scale (∼1 L/h) 129Xe polarizer for clinical, preclinical, and materials NMR and MRI research. Automated and composed mostly of off-the-shelf components, this “hyperpolarizer” is designed to be readily implementable in other laboratories. The device runs with high resonant photon flux (up to 200 W at the Rb D1 line) in the xenon-rich regime (up to 1,800 torr Xe in 500 cc) in either single-batch or stopped-flow mode, negating in part the usual requirement of Xe cryocollection. Excellent agreement is observed among four independent methods used to measure spin polarization. In-cell PXe values of ∼90%, ∼57%, ∼50%, and ∼30% have been measured for Xe loadings of ∼300, ∼500, ∼760, and ∼1,570 torr, respectively. PXe values of ∼41% and ∼28% (with ∼760 and ∼1,545 torr Xe loadings) have been measured after transfer to Tedlar bags and transport to a clinical 3 T scanner for MR imaging, including demonstration of lung MRI with a healthy human subject. Long “in-bag” 129Xe polarization decay times have been measured (T1 ∼38 min and ∼5.9 h at ∼1.5 mT and 3 T, respectively)—more than sufficient for a variety of applications. PMID:23946420

  14. The BRAF inhibitor vemurafenib activates mitochondrial metabolism and inhibits hyperpolarized pyruvate-lactate exchange in BRAF mutant human melanoma cells

    PubMed Central

    Delgado-Goni, Teresa; Falck Miniotis, Maria; Wantuch, Slawomir; Parkes, Harold G.; Marais, Richard; Workman, Paul; Leach, Martin O.; Beloueche-Babari, Mounia

    2016-01-01

    Understanding the impact of BRAF signaling inhibition in human melanoma on key disease mechanisms is important for developing biomarkers of therapeutic response and combination strategies to improve long term disease control. This work investigates the downstream metabolic consequences of BRAF inhibition with vemurafenib, the molecular and biochemical processes that underpin them, their significance for antineoplastic activity and potential as non-invasive imaging response biomarkers.1H NMR spectroscopy showed that vemurafenib decreases the glycolytic activity of BRAF mutant (WM266.4 and SKMEL28) but not BRAFWT (CHL-1 and D04) human melanoma cells. In WM266.4 cells, this was associated with increased acetate, glycine and myo-inositol levels and decreased fatty acyl signals, while the bioenergetic status was maintained. 13C NMR metabolic flux analysis of treated WM266.4 cells revealed inhibition of de novo lactate synthesis and glucose utilization, associated with increased oxidative and anaplerotic pyruvate carboxylase mitochondrial metabolism and decreased lipid synthesis. This metabolic shift was associated with depletion of HKII, acyl-CoA dehydrogenase 9, 3-phosphoglycerate dehydrogenase and monocarboxylate transporter (MCT) 1 and 4 in BRAF mutant but not BRAFWT cells and, interestingly, decreased BRAF mutant cell dependency on glucose and glutamine for growth. Further, the reduction in MCT1 expression observed led to inhibition of hyperpolarized 13C-pyruvate-lactate exchange, a parameter that is translatable to in vivo imaging studies, in live WM266.4 cells. In conclusion, our data provide new insights into the molecular and metabolic consequences of BRAF inhibition in BRAF-driven human melanoma cells that may have potential for combinatorial therapeutic targeting as well as non-invasive imaging of response. PMID:27765851

  15. Real-time cardiac metabolism assessed with hyperpolarized [1-13C]acetate in a large-animal model

    PubMed Central

    Flori, Alessandra; Liserani, Matteo; Frijia, Francesca; Giovannetti, Giulio; Lionetti, Vincenzo; Casieri, Valentina; Positano, Vincenzo; Aquaro, Giovanni Donato; Recchia, Fabio A.; Santarelli, Maria Filomena; Landini, Luigi; Ardenkjaer-Larsen, Jan Henrik; Menichetti, Luca

    2014-01-01

    Dissolution-Dynamic Nuclear Polarization (dissolution-DNP) for Magnetic Resonance (MR) Spectroscopic Imaging has recently emerged as a novel technique for non invasive studies of the metabolic fate of biomolecules in vivo. Since acetate is the most abundant extra- and intra-cellular short-chain fatty acid, we focused on [1-13C]acetate as a promising candidate for a chemical probe to study myocardial metabolism of beating heart. Dissolution-DNP procedure of Na[1-13C]acetate for in vivo cardiac applications with 3T MR scanner was optimized in pigs during bolus injection of doses up to 3 mmoles. The Na[1-13C]acetate formulation was characterized by a liquid-state polarization of 14.2% and T1Eff in vivo of 17.6 ± 1.7 s. In vivo Na[1-13C]acetate kinetic displayed a bimodal shape: [1-13C]acetyl carnitine (AcC) was detected in a slice covering the cardiac volume, and the signal of 13C-acetate and 13C-AcC was modeled using the total Area Under the Curve (AUC) for kinetic analysis. A good correlation was found between the ratio AUC(AcC)/AUC(acetate) and the apparent kinetic constant of metabolic conversion kAcC/r1 from [1-13C]acetate to [1-13C]AcC. Our study proved the feasibility and limitations of administration of large doses of hyperpolarized [1-13C]acetate with dissolution DNP to study by MR spectroscopy the myocardial conversion of [1-13C]acetate in [1-13C]acetyl-carnitine generated by acetyltransferase in healthy pigs. PMID:25201079

  16. Two and three-dimensional segmentation of hyperpolarized 3He magnetic resonance imaging of pulmonary gas distribution

    NASA Astrophysics Data System (ADS)

    Heydarian, Mohammadreza; Kirby, Miranda; Wheatley, Andrew; Fenster, Aaron; Parraga, Grace

    2012-03-01

    A semi-automated method for generating hyperpolarized helium-3 (3He) measurements of individual slice (2D) or whole lung (3D) gas distribution was developed. 3He MRI functional images were segmented using two-dimensional (2D) and three-dimensional (3D) hierarchical K-means clustering of the 3He MRI signal and in addition a seeded region-growing algorithm was employed for segmentation of the 1H MRI thoracic cavity volume. 3He MRI pulmonary function measurements were generated following two-dimensional landmark-based non-rigid registration of the 3He and 1H pulmonary images. We applied this method to MRI of healthy subjects and subjects with chronic obstructive lung disease (COPD). The results of hierarchical K-means 2D and 3D segmentation were compared to an expert observer's manual segmentation results using linear regression, Pearson correlations and the Dice similarity coefficient. 2D hierarchical K-means segmentation of ventilation volume (VV) and ventilation defect volume (VDV) was strongly and significantly correlated with manual measurements (VV: r=0.98, p<.0001 VDV: r=0.97, p<.0001) and mean Dice coefficients were greater than 92% for all subjects. 3D hierarchical K-means segmentation of VV and VDV was also strongly and significantly correlated with manual measurements (VV: r=0.98, p<.0001 VDV: r=0.64, p<.0001) and the mean Dice coefficients were greater than 91% for all subjects. Both 2D and 3D semi-automated segmentation of 3He MRI gas distribution provides a way to generate novel pulmonary function measurements.

  17. Metabolic Response of Prostate Cancer to Nicotinamide Phophoribosyltransferase Inhibition in a Hyperpolarized MR/PET Compatible Bioreactor

    PubMed Central

    Keshari, Kayvan R.; Wilson, David M.; Van Criekinge, Mark; Sriram, Renuka; Koelsch, Bertram L.; Wang, Zhen J.; VanBrocklin, Henry F.; Peehl, Donna M.; O’Brien, Tom; Sampath, Deepak; Carano, Richard A. D.; Kurhanewicz, John

    2015-01-01

    Background Metabolic shifts in disease are of great interest for the development of novel therapeutics. In cancer treatment, these therapies exploit the metabolic phenotype associated with oncogenesis and cancer progression. One recent strategy involves the depletion of the cofactors needed to maintain the high rate of glycolysis seen with the Warburg effect. Specifically, blocking nicotinamide adenine dinucleotide (NAD) biosynthesis via nicotinamide phosphoribosyltransferase (NAMPT) inhibition depletes cancer cells of the NAD needed for glycolysis. To characterize this metabolic phenotype in vivo and describe changes in flux with treatment, non-invasive biomarkers are necessary. One such biomarker is hyperpolarized (HP) [1-13C] pyruvate, a clinically translatable probe that allows real-time assessment of metabolism. Methods We therefore developed a cell perfusion system compatible with HP magnetic resonance (MR) and positron emission tomography (PET) to develop translatable biomarkers of response to NAMPT inhibition in reduced volume cell cultures. Results Using this platform, we observed a reduction in pyruvate flux through lactate dehydrogenase with NAMPT inhibition in prostate cancer cells, and showed that both HP lactate and 2-[18F] fluoro-2-deoxy-D-glucose (FDG) can be used as biomarkers for treatment response of such targeted agents. Moreover, we observed dynamic flux changes whereby HP pyruvate was re-routed to alanine, providing both positive and negative indicators of treatment response. Conclusions This study demonstrated the feasibility of a MR/PET compatible bioreactor approach to efficiently explore cell and tissue metabolism, the understanding of which is critical for developing clinically translatable biomarkers of disease states and responses to therapeutics. PMID:26177608

  18. Learning-induced reduction in post-burst after-hyperpolarization (AHP) is mediated by activation of PKC.

    PubMed

    Seroussi, Yaron; Brosh, Inbar; Barkai, Edi

    2002-09-01

    We studied the role of protein kinase C (PKC) and protein kinase A (PKA) in mediating learning-related long lasting reduction of the post-burst after-hyperpolarization (AHP) in cortical pyramidal neurons. We have shown previously that pyramidal neurons in the rat piriform (olfactory) cortex from trained (TR) rats have reduced post-burst AHP for 3 days after odour-discrimination learning, and that this reduction is due to decreased conductance of calcium-dependent potassium current. In the present study, we examined whether this long-lasting reduction in AHP is mediated by second messenger systems. The broad-spectrum kinase inhibitor, H7, increased the AHP in neurons from TR rats, but not in neurons from pseudo-trained (pseudo-TR) and naive rats. Consequently, the difference in AHP amplitude between neurons from TR and control animals was diminished. This effect was also obtained by application of the specific PKC inhibitor, GF-109203x. The PKC activator, 1-Oleoyl-2-acetyl-sn-glycerol (OAG), significantly reduced the AHP in neurons from naive and pseudo-TR rats, but not in neurons from TR rats, so that the difference between the groups was abolished. The PKA-specific inhibitor, H-89, increased the AHP in neurons from all groups to a similar extent, and the difference in AHP amplitude between neurons from TR rats and neurons from controls was maintained. We suggest that while the post-burst AHP in piriform cortex pyramidal neurons is modulated by both PKC and PKA, a PKC-dependent process maintains the learning-related reduction of the AHP in these cells.

  19. The role of sodium pump activity in the hyperpolarization and in subsequent depolarization of smooth muscle in response to stimulation of post-synaptic alpha 1-adrenoceptors.

    PubMed

    Török, T L; Vizi, E S

    1980-01-01

    The electrical and mechanical activities of guinea pig taenia coli smooth muscle were measured by a "sucrose gap" technique. Under the same experimental conditions the ionic content of smooth muscle was also measured. The mean value of the resting potential was 56.9 +/- 1.1 mV (S.E.M.; n = 46). In normal Krebs solution immediately after dissection intracellular sodium amounted to 30.1, and intracellular calcium to 1.5 mmole x kg-1 wet weight. In response to adrenaline administration there was a Ca-dependent hyperpolarization (peak, 6.8 +/- 0.3 mV S.E.M.; n = 5) and an increased Na efflux with a rate constant (k) of 0.16 min-1 (60'). Removal of adrenaline was followed by so-called "postadrenaline depolarization" i.e. the decrease of the membrane potential was greater than the initial rise, an effect enhanced by ouabain (2 X 10(-5) M). Clonidine (5.3 X 10(-6) M), a selective presynaptic-adrenoceptor (alpha 2-receptor) stimulant failed to produce hyperpolarization, however, phenylephrine (5 X 10(-5) M) a pure postsynaptic alpha-adrenoceptor (alpha 1-receptor) stimulant produced a similar effect as adrenaline. In addition, yohimbine (1.4 X 10(-6) M), a typical presynaptic alpha-adrenoceptor inhibitor failed to affect the action of adrenaline or phenylephrine. These facts indicate that the alpha 1-adrenoceptors present on the smooth muscle are different from those situated presynaptically on the cholinergic nerve terminals modulating the release of acetylcholine. The effect of ouabain to lower membrane potential proved to be Ca2+-dependent. The intracellular sodium content was enhanced by ouabain from 30.1 to 90.9 +/- 4.7 mmole x kg-1 wet weight (60'). On washing out ouabain, hyperpolarization "post-ouabain hyperpolarization" was detected, i.e. the rise of membrane potential was greater than the initial fall. It is suggested that the sodium pump plays a significant role in the post-ouabain hyperpolarization. Direct calculation of sodium movements suggests that the

  20. 13C magnetic resonance spectroscopy measurements with hyperpolarized [1‐13C] pyruvate can be used to detect the expression of transgenic pyruvate decarboxylase activity in vivo

    PubMed Central

    Dzien, Piotr; Tee, Sui‐Seng; Kettunen, Mikko I.; Lyons, Scott K.; Larkin, Timothy J.; Timm, Kerstin N.; Hu, De‐En; Wright, Alan; Rodrigues, Tiago B.; Serrao, Eva M.; Marco‐Rius, Irene; Mannion, Elizabeth; D'Santos, Paula; Kennedy, Brett W. C.

    2015-01-01

    Purpose Dissolution dynamic nuclear polarization can increase the sensitivity of the 13C magnetic resonance spectroscopy experiment by at least four orders of magnitude and offers a novel approach to the development of MRI gene reporters based on enzymes that metabolize 13C‐labeled tracers. We describe here a gene reporter based on the enzyme pyruvate decarboxylase (EC 4.1.1.1), which catalyzes the decarboxylation of pyruvate to produce acetaldehyde and carbon dioxide. Methods Pyruvate decarboxylase from Zymomonas mobilis (zmPDC) and a mutant that lacked enzyme activity were expressed using an inducible promoter in human embryonic kidney (HEK293T) cells. Enzyme activity was measured in the cells and in xenografts derived from the cells using 13C MRS measurements of the conversion of hyperpolarized [1‐13C] pyruvate to H13 CO3–. Results Induction of zmPDC expression in the cells and in the xenografts derived from them resulted in an approximately two‐fold increase in the H13 CO3–/[1‐13C] pyruvate signal ratio following intravenous injection of hyperpolarized [1‐13C] pyruvate. Conclusion We have demonstrated the feasibility of using zmPDC as an in vivo reporter gene for use with hyperpolarized 13C MRS. Magn Reson Med 76:391–401, 2016. © 2015 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. PMID:26388418

  1. Inhibitory actions of ZENECA ZD7288 on whole-cell hyperpolarization activated inward current (If) in guinea-pig dissociated sinoatrial node cells.

    PubMed Central

    BoSmith, R. E.; Briggs, I.; Sturgess, N. C.

    1993-01-01

    1. ZENECA ZD7288 (4-(N-ethyl-N-phenylamino)-1,2-dimethyl-6-(methylamino) pyridinium chloride) is a sinoatrial node (SAN) modulating agent which produces a selective slowing of the heart rate. Its effects have been studied in single, freshly dissociated guinea-pig SAN cells, by standard patch clamp procedures. 2. Whole-cell inward currents were evoked by hyperpolarizing voltage clamp steps from a holding potential of -40 mV. ZD7288 inhibited the hyperpolarization activated cationic current (If) in a concentration-dependent manner. The 'selective bradycardic agents' alinidine and UL-FS 49 (zatebradine) both also inhibited If. 3. The activation of If was investigated by measuring tail current amplitudes at +20 mV after hyperpolarizing steps to different potentials to activate the current. The reduction in If resulted from both a shift in the If current activation curve in the negative direction on the voltage axis, and also a reduction in the activation curve amplitude. 4. ZD7288 did not affect the ion selectivity of the If channel, since the tail current reversal potential was unchanged in the presence of the drug. 5. With ZD7288 the inhibition of If was not use-dependent, whereas UL-FS 49 displayed use-dependence in the block of the If current. 6. Whereas ZD7288 had no significant effect on the delayed rectifier current (Ik) in these cells, both alinidine and UL-FS 49 significantly reduced Ik at the same concentrations which reduced If. 7. The data show that ZD7288 reduces If by affecting the activation characteristics of the If current; this inhibition may account for this agent's selective bradycardiac properties. PMID:7693281

  2. The magnitudes of hyperpolarization-activated and low-voltage-activated potassium currents co-vary in neurons of the ventral cochlear nucleus.

    PubMed

    Cao, Xiao-Jie; Oertel, Donata

    2011-08-01

    In the ventral cochlear nucleus (VCN), neurons have hyperpolarization-activated conductances, which in some cells are enormous, that contribute to the ability of neurons to convey acoustic information in the timing of their firing by decreasing the input resistance and speeding-up voltage changes. Comparisons of the electrophysiological properties of neurons in the VCN of mutant mice that lack the hyperpolarization-activated cyclic nucleotide-gated channel α subunit 1 (HCN1(-/-)) (Nolan et al. 2003) with wild-type controls (HCN1(+/+)) and with outbred ICR mice reveal that octopus, T stellate, and bushy cells maintain their electrophysiological distinctions in all strains. Hyperpolarization-activated (I(h)) currents were smaller and slower, input resistances were higher, and membrane time constants were longer in HCN1(-/-) than in HCN1(+/+) in octopus, bushy, and T stellate cells. There were significant differences in the average magnitudes of I(h), input resistances, and time constants between HCN1(+/+) and ICR mice, but the resting potentials did not differ between strains. I(h) is opposed by a low-voltage-activated potassium (I(KL)) current in bushy and octopus cells, whose magnitudes varied widely between neuronal types and between strains. The magnitudes of I(h) and I(KL) were correlated across neuronal types and across mouse strains. Furthermore, these currents balanced one another at the resting potential in individual cells. The magnitude of I(h) and I(KL) is linked in bushy and octopus cells and varies not only between HCN1(-/-) and HCN1(+/+) but also between "wild-type" strains of mice, raising the question to what extent the wild-type strains reflect normal mice.

  3. Persistent Mitochondrial Hyperpolarization, Increased Reactive Oxygen Intermediate Production, and Cytoplasmic Alkalinization Characterize Altered IL-10 Signaling in Patients with Systemic Lupus Erythematosus1

    PubMed Central

    Gergely, Peter; Niland, Brian; Gonchoroff, Nick; Pullmann, Rudolf; Phillips, Paul E.; Perl, Andras

    2014-01-01

    Abnormal death signaling in lymphocytes of systemic lupus erythematosus (SLE) patients has been associated with elevation of the mitochondrial transmembrane potential (Δψm) and increased production of reactive oxygen intermediates (ROI). The resultant ATP depletion sensitizes T cells for necrosis that may significantly contribute to inflammation in patients with SLE. In the present study, the role of mitochondrial signal processing in T cell activation was investigated. CD3/CD28 costimulation of PBL elicited transient mitochondrial hyperpolarization and intracellular pH (pHi) elevation, followed by increased ROI production. Baseline Δψm, ROI production, and pHi were elevated, while T cell activation-induced changes were blunted in 15 patients with SLE in comparison with 10 healthy donors and 10 rheumatoid arthritis patients. Similar to CD3/CD28 costimulation, treatment of control PBL with IL-3, IL-10, TGF-β1, and IFN-γ led to transient Δψm elevation. IL-10 had diametrically opposing effects on mitochondrial signaling in lupus and control donors. Unlike healthy or rheumatoid arthritis PBL, cells of lupus patients were resistant to IL-10-induced mitochondrial hyperpolarization. By contrast, IL-10 enhanced ROI production and cell death in lupus PBL without affecting ROI levels and survival of control PBL. Ab-mediated IL-10 blockade or stimulation with antagonistic lymphokine IL-12 normalized baseline and CD3/CD28-induced changes in ROI production and pHi with no impact on Δψm of lupus PBL. The results suggest that mitochondrial hyperpolarization, increased ROI production, and cytoplasmic alkalinization play crucial roles in altered IL-10 responsiveness in SLE. PMID:12097418

  4. Modulation of single hyperpolarization-activated channels (i(f)) by cAMP in the rabbit sino-atrial node.

    PubMed Central

    DiFrancesco, D; Mangoni, M

    1994-01-01

    1. The hyperpolarization-activated 'pacemaker' current (i(f)) was recorded in inside-out patches excised from rabbit sino-atrial (SA) node cell membranes. 2. Single-channel activity could be resolved in patches containing only a few channels; the voltage dependence of single-channel size and single-channel conductance (0.97 pS) were similar to those measured previously in cell-attached conditions. 3. Perfusion of the intracellular side of the patch membrane with 10 microM cAMP facilitated the opening of single i(f) channels on hyperpolarization. The cAMP-induced i(f) current activation occurred without modification of the single-channel conductance. 4. Modification by cAMP of the probability of channel opening was investigated with respect to the latency to first opening during hyperpolarization and in patches containing a large number of channels (macro-patches). First-latency histograms showed that cAMP shifts the probability curve of first openings to shorter times, in agreement with a cAMP-induced facilitation of channel opening. In macro-patches, measurement of the voltage dependence of the open probability by a slow voltage ramp protocol showed that cAMP shifts the probability curve to more positive voltages without modifying its shape. 5. In cell-free macro-patches the normalized open probability curve in control solutions was centred around -121.9 mV, a voltage some 30 mV more negative than in cell-attached macro-patches. Negative shifting of the curve after patch excision could only partly be explained by the removal of intracellular cAMP, and progressed with time during the ramp protocol, suggesting the presence of a run-down process independent from cAMP. PMID:7516974

  5. Accelerated Ca2+ entry by membrane hyperpolarization due to Ca2+-activated K+ channel activation in response to histamine in chondrocytes.

    PubMed

    Funabashi, Kenji; Ohya, Susumu; Yamamura, Hisao; Hatano, Noriyuki; Muraki, Katsuhiko; Giles, Wayne; Imaizumi, Yuji

    2010-04-01

    In articular cartilage inflammation, histamine release from mast cells is a key event. It can enhance cytokine production and matrix synthesis and also promote cell proliferation by stimulating chondrocytes. In this study, the functional impact of Ca(2+)-activated K(+) (K(Ca)) channels in the regulation of intracellular Ca(2+) concentration ([Ca(2+)](i)) in chondrocytes in response to histamine was examined using OUMS-27 cells, as a model of chondrocytes derived from human chondrosarcoma. Application of histamine induced a significant [Ca(2+)](i) rise and also membrane hyperpolarization, and both effects were mediated by the stimulation of H(1) receptors. The histamine-induced membrane hyperpolarization was attenuated to approximately 50% by large-conductance K(Ca) (BK) channel blockers, and further reduced by intermediate (IK) and small conductance K(Ca) (SK) channel blockers. The tonic component of histamine-induced [Ca(2+)](i) rise strongly depended on the presence of extracellular Ca(2+) ([Ca(2+)](o)) and was markedly reduced by La(3+) or Gd(3+) but not by nifedipine. It was significantly attenuated by BK channel blockers, and further blocked by the cocktail of BK, IK, and SK channel blockers. The K(Ca) blocker cocktail also significantly reduced the store-operated Ca(2+) entry (SOCE), which was induced by Ca(2+) addition after store-depletion by thapsigargin in [Ca(2+)](o) free solution. Our results demonstrate that the histamine-induced membrane hyperpolarization in chondrocytes due to K(Ca) channel activation contributes to sustained Ca(2+) entry mainly through SOCE channels in OUMS-27 cells. Thus, K(Ca) channels appear to play an important role in the positive feedback mechanism of [Ca(2+)](i) regulation in chondrocytes in the presence of articular cartilage inflammation.

  6. The magnitudes of hyperpolarization-activated and low-voltage-activated potassium currents co-vary in neurons of the ventral cochlear nucleus

    PubMed Central

    Cao, Xiao-Jie

    2011-01-01

    In the ventral cochlear nucleus (VCN), neurons have hyperpolarization-activated conductances, which in some cells are enormous, that contribute to the ability of neurons to convey acoustic information in the timing of their firing by decreasing the input resistance and speeding-up voltage changes. Comparisons of the electrophysiological properties of neurons in the VCN of mutant mice that lack the hyperpolarization-activated cyclic nucleotide-gated channel α subunit 1 (HCN1−/−) (Nolan et al. 2003) with wild-type controls (HCN1+/+) and with outbred ICR mice reveal that octopus, T stellate, and bushy cells maintain their electrophysiological distinctions in all strains. Hyperpolarization-activated (Ih) currents were smaller and slower, input resistances were higher, and membrane time constants were longer in HCN1−/− than in HCN1+/+ in octopus, bushy, and T stellate cells. There were significant differences in the average magnitudes of Ih, input resistances, and time constants between HCN1+/+ and ICR mice, but the resting potentials did not differ between strains. Ih is opposed by a low-voltage-activated potassium (IKL) current in bushy and octopus cells, whose magnitudes varied widely between neuronal types and between strains. The magnitudes of Ih and IKL were correlated across neuronal types and across mouse strains. Furthermore, these currents balanced one another at the resting potential in individual cells. The magnitude of Ih and IKL is linked in bushy and octopus cells and varies not only between HCN1−/− and HCN1+/+ but also between “wild-type” strains of mice, raising the question to what extent the wild-type strains reflect normal mice. PMID:21562186

  7. Assessing Oxidative Stress in Tumors by Measuring the Rate of Hyperpolarized [1-13C]Dehydroascorbic Acid Reduction Using 13C Magnetic Resonance Spectroscopy*

    PubMed Central

    Timm, Kerstin N.; Hu, De-En; Williams, Michael; Wright, Alan J.; Kettunen, Mikko I.; Kennedy, Brett W. C.; Larkin, Timothy J.; Dzien, Piotr; Marco-Rius, Irene; Bohndiek, Sarah E.; Brindle, Kevin M.

    2017-01-01

    Rapid cancer cell proliferation promotes the production of reducing equivalents, which counteract the effects of relatively high levels of reactive oxygen species. Reactive oxygen species levels increase in response to chemotherapy and cell death, whereas an increase in antioxidant capacity can confer resistance to chemotherapy and is associated with an aggressive tumor phenotype. The pentose phosphate pathway is a major site of NADPH production in the cell, which is used to maintain the main intracellular antioxidant, glutathione, in its reduced state. Previous studies have shown that the rate of hyperpolarized [1-13C]dehydroascorbic acid (DHA) reduction, which can be measured in vivo using non-invasive 13C magnetic resonance spectroscopic imaging, is increased in tumors and that this is correlated with the levels of reduced glutathione. We show here that the rate of hyperpolarized [1-13C]DHA reduction is increased in tumors that have been oxidatively prestressed by depleting the glutathione pool by buthionine sulfoximine treatment. This increase was associated with a corresponding increase in pentose phosphate pathway flux, assessed using 13C-labeled glucose, and an increase in glutaredoxin activity, which catalyzes the glutathione-dependent reduction of DHA. These results show that the rate of DHA reduction depends not only on the level of reduced glutathione, but also on the rate of NADPH production, contradicting the conclusions of some previous studies. Hyperpolarized [1-13C]DHA can be used, therefore, to assess the capacity of tumor cells to resist oxidative stress in vivo. However, DHA administration resulted in transient respiratory arrest and cardiac depression, which may prevent translation to the clinic. PMID:27994059

  8. Carbonic Anhydrase Activity Monitored In Vivo by Hyperpolarized 13C-Magnetic Resonance Spectroscopy Demonstrates Its Importance for pH Regulation in Tumors.

    PubMed

    Gallagher, Ferdia A; Sladen, Helen; Kettunen, Mikko I; Serrao, Eva M; Rodrigues, Tiago B; Wright, Alan; Gill, Andrew B; McGuire, Sarah; Booth, Thomas C; Boren, Joan; McIntyre, Alan; Miller, Jodi L; Lee, Shen-Han; Honess, Davina; Day, Sam E; Hu, De-En; Howat, William J; Harris, Adrian L; Brindle, Kevin M

    2015-10-01

    Carbonic anhydrase buffers tissue pH by catalyzing the rapid interconversion of carbon dioxide (CO2) and bicarbonate (HCO3 (-)). We assessed the functional activity of CAIX in two colorectal tumor models, expressing different levels of the enzyme, by measuring the rate of exchange of hyperpolarized (13)C label between bicarbonate (H(13)CO3(-)) and carbon dioxide ((13)CO2), following injection of hyperpolarized H(13)CO3(-), using (13)C-magnetic resonance spectroscopy ((13)C-MRS) magnetization transfer measurements. (31)P-MRS measurements of the chemical shift of the pH probe, 3-aminopropylphosphonate, and (13)C-MRS measurements of the H(13)CO3(-)/(13)CO2 peak intensity ratio showed that CAIX overexpression lowered extracellular pH in these tumors. However, the (13)C measurements overestimated pH due to incomplete equilibration of the hyperpolarized (13)C label between the H(13)CO3(-) and (13)CO2 pools. Paradoxically, tumors overexpressing CAIX showed lower enzyme activity using magnetization transfer measurements, which can be explained by the more acidic extracellular pH in these tumors and the decreased activity of the enzyme at low pH. This explanation was confirmed by administration of bicarbonate in the drinking water, which elevated tumor extracellular pH and restored enzyme activity to control levels. These results suggest that CAIX expression is increased in hypoxia to compensate for the decrease in its activity produced by a low extracellular pH and supports the hypothesis that a major function of CAIX is to lower the extracellular pH.

  9. Carbonic anhydrase activity monitored in vivo by hyperpolarized 13C-magnetic resonance spectroscopy demonstrate its importance for pH regulation in tumors

    PubMed Central

    Gallagher, Ferdia A.; Sladen, Helen; Kettunen, Mikko I.; Serrao, Eva M.; Rodrigues, Tiago B.; Wright, Alan; Gill, Andrew B.; McGuire, Sarah; Booth, Thomas C.; Boren, Joan; McIntyre, Alan; Miller, Jodi L.; Lee, Shen-Han; Honess, Davina; Day, Sam E.; Hu, De-en; Howat, William J.; Harris, Adrian L.; Brindle, Kevin M.

    2015-01-01

    Carbonic anhydrase (CA) buffers tissue pH by catalyzing the rapid interconversion of carbon dioxide (CO2) and bicarbonate (HCO3−). We assessed the functional activity of CAIX in two colorectal tumor models, expressing different levels of the enzyme, by measuring the rate of exchange of hyperpolarized 13C label between bicarbonate (H13CO3−) and carbon dioxide (13CO2), following injection of hyperpolarized H13CO3−, using 13C magnetic resonance spectroscopy (13C-MRS) magnetization transfer measurements. 31P-MRS measurements of the chemical shift of the pH probe, 3-aminopropylphosphonate, and 13C-MRS measurements of the H13CO3−/13CO2 peak intensity ratio showed that CAIX overexpression lowered extracellular pH in these tumors. However, the 13C measurements overestimated pH due to incomplete equilibration of the hyperpolarized 13C label between the H13CO3− and 13CO2 pools. Paradoxically, tumors overexpressing CAIX showed lower enzyme activity using magnetization transfer measurements, which can be explained by the more acidic extracellular pH in these tumors and the decreased activity of the enzyme at low pH. This explanation was confirmed by administration of bicarbonate in the drinking water, which elevates tumor extracellular pH and restored enzyme activity to control levels. These results suggest that CAIX expression is increased in hypoxia to compensate for the decrease in its activity produced by a low extracellular pH, and supports the hypothesis that a major function of CAIX is to lower the extracellular pH. PMID:26249175

  10. Hyperpolarization-activated current Ih in mouse trigeminal sensory neurons in a transgenic mouse model of familial hemiplegic migraine type-1.

    PubMed

    Eroli, Francesca; Vilotti, Sandra; van den Maagdenberg, Arn M J M; Nistri, Andrea

    2017-03-29

    Transgenic knock-in (KI) mice that express CaV2.1 channels containing an R192Q gain-of-function mutation in the α1A subunit known to cause familial hemiplegic migraine type-1 in patients, exhibit key disease characteristics and provide a useful tool to investigate pathophysiological mechanisms of pain transduction. Previously, KI trigeminal sensory neurons were shown to exhibit constitutive hyperexcitability due to up-regulation of ATP-gated P2X3 receptors that trigger spike activity at a more negative threshold. This implies that intrinsic neuronal conductances may shape action potential generation in response to ATP, which could act as a mediator of migraine headache. Here we investigated whether the hyperpolarization-activated conductance Ih, mediated by hyperpolarization-activated cyclic nucleotide-gated channel (HCN) channels, contributes to sub-threshold behavior and firing in wild-type (WT) and KI trigeminal ganglia (TG) neurons. Whereas most WT and KI trigeminal neurons expressed Ih current, blocked by the specific inhibitor ZD7288, it was smaller in KI neurons despite similar activation and deactivation kinetics. HCN1 and HCN2 were the most abundantly expressed subunits in TG, both in situ and in culture. In KI TG neurons, HCN2 subunits were predominantly present in the cytoplasm, not at the plasma membrane, likely accounting for the smaller Ih of such cells. ZD7288 hyperpolarized the membrane potential, thereby raising the firing threshold, and prolonging the spike trajectory to generate fewer spikes due to P2X3 receptor activation. The low amplitude of Ih in KI TG neurons suggests that down-regulation of Ih current in sub-threshold behavior acts as a compensatory mechanism to limit sensory hyperexcitability, manifested under certain stressful stimuli.

  11. Ratiometric analysis in hyperpolarized NMR (I): test of the two-site exchange model and the quantification of reaction rate constants.

    PubMed

    Li, Lin Z; Kadlececk, Stephen; Xu, He N; Daye, Dania; Pullinger, Benjamin; Profka, Harrilla; Chodosh, Lewis; Rizi, Rahim

    2013-10-01

    Conventional methods for the analysis of in vivo hyperpolarized (13) C NMR data from the lactate dehydrogenase (LDH) reaction usually make assumptions on the stability of rate constants and/or the validity of the two-site exchange model. In this study, we developed a framework to test the validity of the assumption of stable reaction rate constants and the two-site exchange model in vivo via ratiometric fitting of the time courses of the signal ratio L(t)/P(t). Our analysis provided evidence that the LDH enzymatic kinetics observed by hyperpolarized NMR are in near-equilibrium and satisfy the two-site exchange model for only a specific time window. In addition, we quantified both the forward and reverse exchange rate constants of the LDH reaction for the transgenic and mouse xenograft models of breast cancer using the ratio fitting method developed, which includes only two modeling parameters and is less sensitive to the influence of instrument settings/protocols, such as flip angles, degree of polarization and tracer dosage. We further compared the ratio fitting method with a conventional two-site exchange modeling method, i.e. the differential equation fitting method, using both the experimental and simulated hyperpolarized NMR data. The ratio fitting method appeared to fit better than the differential equation fitting method for the reverse rate constant on the mouse tumor data, with less relative errors on average, whereas the differential equation fitting method also resulted in a negative reverse rate constant for one tumor. The simulation results indicated that the accuracy of both methods depends on the width of the transport function, noise level and rate constant ratio; one method may be more accurate than the other based on the experimental/biological conditions aforementioned. We were able to categorize our tumor models into specific conditions of the computer simulation and to estimate the errors of rate quantification. We also discussed possible

  12. Magnetic Field Strength Dependence of Transverse Relaxation and Signal-to-Noise Ratio for Hyperpolarized Xenon-129 and Helium-3 Gas Magnetic Resonance Imaging of Lungs

    NASA Astrophysics Data System (ADS)

    Dominguez-Viqueira, William

    Magnetic resonance (MR) imaging with hyperpolarized noble gases (HNG), 3He or 129Xe, has become a promising approach for studying lung anatomy and function. Unlike conventional MR imaging, the magnetization in HNG MR is independent of the magnetic field strength. This means that no improvement in signal-to-noise ratio (SNR) is expected with increasing clinical field strength above ˜0.25T. Furthermore, it has been predicted that the SNR may decline at clinical field strength due to decreases in the apparent transverse relaxation time (T2*), caused by the increased magnetic susceptibility induced field gradients at the air-tissue interface. In this thesis the magnetic field strength dependence of T2* and SNR in HNG MR is investigated experimentally in rodent and human lungs. For rodent imaging, a novel broad-band (0.1-100MHz) variable field strength MR imaging system for rodents was built. This system permitted imaging of 129Xe, 3He and 1H at low magnetic field strengths (3-73.5mT) to experimentally investigate the field dependence of HNG imaging SNR in rodent lungs. In vivo 129Xe and 3He signals were acquired at 73.5mT and T 2* was estimated to be approximately 180+/-8 ms, in good agreement with previously reported values. At 73.5mT, image noise is dominated by losses originated from the radiofrequency (RF) coils. To address this issue, RF coils were built using different types of copper wire and compared in phantoms and in vivo in rat lungs using hyperpolarized 3He and 129Xe gas. An SNR improvement of up to 200% was obtained with Litz wire compared to conventional copper wire. This improvement demonstrated the feasibility of HNG lung imaging in rodents at 73.5mT with SNR comparable to that obtained at clinical field strengths. To verify the SNR field dependence in humans, hyperpolarized 3He lung imaging at two commonly used clinical field strengths (1.5T and 3T) was performed in the same volunteers and compared. No significant differences in SNR were obtained

  13. Low-field thermal mixing in [1-(13)C] pyruvic acid for brute-force hyperpolarization.

    PubMed

    Peat, David T; Hirsch, Matthew L; Gadian, David G; Horsewill, Anthony J; Owers-Bradley, John R; Kempf, James G

    2016-07-28

    We detail the process of low-field thermal mixing (LFTM) between (1)H and (13)C nuclei in neat [1-(13)C] pyruvic acid at cryogenic temperatures (4-15 K). Using fast-field-cycling NMR, (1)H nuclei in the molecule were polarized at modest high field (2 T) and then equilibrated with (13)C nuclei by fast cycling (∼300-400 ms) to a low field (0-300 G) that activates thermal mixing. The (13)C NMR spectrum was recorded after fast cycling back to 2 T. The (13)C signal derives from (1)H polarization via LFTM, in which the polarized ('cold') proton bath contacts the unpolarised ('hot') (13)C bath at a field so low that Zeeman and dipolar interactions are similar-sized and fluctuations in the latter drive (1)H-(13)C equilibration. By varying mixing time (tmix) and field (Bmix), we determined field-dependent rates of polarization transfer (1/τ) and decay (1/T1m) during mixing. This defines conditions for effective mixing, as utilized in 'brute-force' hyperpolarization of low-γ nuclei like (13)C using Boltzmann polarization from nearby protons. For neat pyruvic acid, near-optimum mixing occurs for tmix∼ 100-300 ms and Bmix∼ 30-60 G. Three forms of frozen neat pyruvic acid were tested: two glassy samples, (one well-deoxygenated, the other O2-exposed) and one sample pre-treated by annealing (also well-deoxygenated). Both annealing and the presence of O2 are known to dramatically alter high-field longitudinal relaxation (T1) of (1)H and (13)C (up to 10(2)-10(3)-fold effects). Here, we found smaller, but still critical factors of ∼(2-5)× on both τ and T1m. Annealed, well-deoxygenated samples exhibit the longest time constants, e.g., τ∼ 30-70 ms and T1m∼ 1-20 s, each growing vs. Bmix. Mixing 'turns off' for Bmix > ∼100 G. That T1m≫τ is consistent with earlier success with polarization transfer from (1)H to (13)C by LFTM.

  14. Expression and properties of hyperpolarization-activated current in rat dorsal root ganglion neurons with known sensory function

    PubMed Central

    Gao, L L; McMullan, S; Djouhri, L; Acosta, C; Harper, A A; Lawson, S N

    2012-01-01

    The hyperpolarization-activated current (Ih) has been implicated in nociception/pain, but its expression levels in nociceptors remained unknown. We recorded Ih magnitude and properties by voltage clamp from dorsal root ganglion (DRG) neurons in vivo, after classifying them as nociceptive or low-threshold-mechanoreceptors (LTMs) and as having C-, Aδ- or Aα/β-conduction velocities (CVs). For both nociceptors and LTMs, Ih amplitude and Ih density (at −100 mV) were significantly positively correlated with CV. Median Ih magnitudes and Ih density in neuronal subgroups were respectively: muscle spindle afferents (MSAs): −4.6 nA, −33 pA pF−1; cutaneous Aα/β LTMs: −2.2 nA, −20 pA pF−1; Aβ-nociceptors: −2.6 nA, −21 pA pF−1; both Aδ-LTMs and nociceptors: −1.3 nA, ∼−14 pA pF−1; C-LTMs: −0.4 nA, −7.6 pA pF−1; and C-nociceptors: −0.26 nA, −5 pA pF−1. Ih activation slow time constants (slow τ values) were strongly correlated with fast τ values; both were shortest in MSAs. Most neurons had τ values consistent with HCN1-related Ih; others had τ values closer to HCN1+HCN2 channels, or HCN2 in the presence of cAMP. In contrast, median half-activation voltages (V0.5) of −80 to −86 mV for neuronal subgroups suggest contributions of HCN2 to Ih. τ values were unrelated to CV but were inversely correlated with Ih and Ih density for all non-MSA LTMs, and for Aδ-nociceptors. From activation curves ∼2–7% of Ih would be activated at normal membrane potentials. The high Ih may be important for excitability of A-nociceptors (responsible for sharp/pricking-type pain) and Aα/β-LTMs (tactile sensations and proprioception). Underlying HCN expression in these subgroups therefore needs to be determined. Altered Ih expression and/or properties (e.g. in chronic/pathological pain states) may influence both nociceptor and LTM excitability. PMID:22753545

  15. MO-G-18C-03: Evaluation of Deformable Image Registration for Lung Motion Estimation Using Hyperpolarized Gas Tagging MRI

    SciTech Connect

    Huang, Q; Zhang, Y; Liu, Y; Hu, L; Yin, F; Cai, J; Miller, W

    2014-06-15

    Purpose: Hyperpolarized gas (HP) tagging MRI is a novel imaging technique for direct measurement of lung motion during breathing. This study aims to quantitatively evaluate the accuracy of deformable image registration (DIR) in lung motion estimation using HP tagging MRI as references. Methods: Three healthy subjects were imaged using the HP MR tagging, as well as a high-resolution 3D proton MR sequence (TrueFISP) at the end-of-inhalation (EOI) and the end-of-exhalation (EOE). Ground truth of lung motion and corresponding displacement vector field (tDVF) was derived from HP tagging MRI by manually tracking the displacement of tagging grids between EOI and EOE. Seven different DIR methods were applied to the high-resolution TrueFISP MR images (EOI and EOE) to generate the DIR-based DVFs (dDVF). The DIR methods include Velocity (VEL), MIM, Mirada, multi-grid B-spline from Elastix (MGB) and 3 other algorithms from DIRART toolbox (Double Force Demons (DFD), Improved Lucas-Kanade (ILK), and Iterative Optical Flow (IOF)). All registrations were performed by independent experts. Target registration error (TRE) was calculated as tDVF – dDVF. Analysis was performed for the entire lungs, and separately for the upper and lower lungs. Results: Significant differences between tDVF and dDVF were observed. Besides the DFD and IOF algorithms, all other dDVFs showed similarity in deformation magnitude distribution but away from the ground truth. The average TRE for entire lung ranged 2.5−23.7mm (mean=8.8mm), depending on the DIR method and subject's breathing amplitude. Larger TRE (13.3–23.7mm) was found in subject with larger breathing amplitude of 45.6mm. TRE was greater in lower lung (2.5−33.9 mm, mean=12.4mm) than that in upper lung (2.5−11.9 mm, mean=5.8mm). Conclusion: Significant differences were observed in lung motion estimation between the HP gas tagging MRI method and the DIR methods, especially when lung motion is large. Large variation among different DIR

  16. Hybrid channel flow-type mechanisms in the Greater Himalayan Sequence (West Nepal): new constraints from vorticity of flow and quartz petrofabric analyses.

    NASA Astrophysics Data System (ADS)

    Frassi, Chiara

    2016-04-01

    Three main tectono-metamorphic units are classically recognized along the Himalayan belt: the Lesser Himalayan (LH), the Greater Himalayan sequence (GHS) and the Tibetan Sedimentary sequence (TSS). The GHS may be interpreted as a low-viscosity tabular body of mid-crustal rocks extruded southward in Miocene times beneath the Tibetan plateau between two parallel and opposite-sense crustal-scale shear zones: the Main Central thrust at the base, and the South Tibetan Detachment system at the top. The pre-/syn-shearing mineral assemblage documented within these crustal-scale shear zones indicates that the metamorphic grade increases toward the core of the GHS producing an inverted and a normal thermal gradient respectively on the top and on the bottom of the slab. In addition, thermal profiles estimated using both petrology- and microstructures/fabrics-based thermometers indicate that the metamorphic isograds are condensed. Although horizontal extension and vorticity estimates collected across the GHS could be strongly biased by the criteria used to define the map position of the MCT, published vorticity data document general shear flow (1>Wk>0) within the slab with a pure-shear component of flow slightly predominant within the core of the GHS whereas the simple-shear component seems to dominate at the top of the slab. The lower boundary of the GHS records a general shear flow with a comparable contribution of simple and pure shearing. The associated crustal extrusion is compatible with Couette - Poiseuille velocity flow profile as assumed in crustal-scale channel flow-type models In this study, the quartz c-axis petrofabrics, vorticity and deformation-temperature studies are integrated with microstructures and metamorphic studies to individuate the location of the MCT and to document the spatial distribution of ductile deformation patterns across the lower portion of the GHS exposed in the Chaudabise river valley in western Nepal. My results indicate that the Main

  17. A Comparison between Radiolabeled Fluorodeoxyglucose Uptake and Hyperpolarized 13C-Labeled Pyruvate Utilization as Methods for Detecting Tumor Response to Treatment12

    PubMed Central

    Witney, Timothy H; Kettunen, Mikko I; Day, Samuel E; Hu, De-en; Neves, Andre A; Gallagher, Ferdia A; Fulton, Sandra M; Brindle, Kevin M

    2009-01-01

    Detection of early tumor responses to treatment can give an indication of clinical outcome. Positron emission tomography measurements of the uptake of the glucose analog, [18F] 2-fluoro-2-deoxy-d-glucose (FDG), have demonstrated their potential for detecting early treatment response in the clinic. We have shown recently that 13C magnetic resonance spectroscopy and spectroscopic imaging measurements of the uptake and conversion of hyperpolarized [1-13C]pyruvate into [1-13C]lactate can be used to detect treatment response in a murine lymphoma model. The present study compares these magnetic resonance measurements with changes in FDG uptake after chemotherapy. A decrease in FDG uptake was found to precede the decrease in flux of hyperpolarized 13C label between pyruvate and lactate, both in tumor cells in vitro and in tumors in vivo. However, the magnitude of the decrease in FDG uptake and the decrease in pyruvate to lactate flux was comparable at 24 hours after drug treatment. In cells, the decrease in FDG uptake was shown to correlate with changes in plasma membrane expression of the facilitative glucose transporters, whereas the decrease in pyruvate to lactate flux could be explained by an increase in poly(ADP-ribose) polymerase activity and subsequent depletion of the NAD(H) pool. These results show that measurement of flux between pyruvate and lactate may be an alternative to FDG-positron emission tomography for imaging tumor treatment response in the clinic. PMID:19484146

  18. DIFFERENTIAL AND AGE-DEPENDENT EXPRESSION OF HYPERPOLARIZATION-ACTIVATED, CYCLIC NUCLEOTIDE-GATED CATION CHANNEL ISOFORMS 1–4 SUGGESTS EVOLVING ROLES IN THE DEVELOPING RAT HIPPOCAMPUS

    PubMed Central

    BENDER, R. A.; BREWSTER, A.; SANTORO, B.; LUDWIG, A.; HOFMANN, F.; BIEL, M.; BARAM, T. Z.

    2011-01-01

    Hyperpolarization-activated cation currents (Ih) are found in several brain regions including thalamus and hippocampus. Important functions of these currents in promoting synchronized network activity and in determining neuronal membrane properties have been progressively recognized, but the molecular underpinnings of these currents are only emerging. Ih currents are generated by hyperpolarization-activated, cyclic nucleotide-gated cation channels (HCNs). These channel proteins are encoded by at least four HCN genes, that govern the kinetic and functional properties of the resulting channels. Because of the potential impact of Ih-mediated coordinated neuronal activity on the maturation of the functional hippocampal network, this study focused on determining the expression of the four members of the HCN gene family throughout postnatal hippocampal development at both the regional and single cell level. The results of these experiments demonstrated that HCNs 1, 2 and 4 are differentially expressed in interneuronal and principal cell populations of the rat hippocampal formation. Expression profiles of each HCN isoform evolve during postnatal development, and patterns observed during early postnatal ages differ significantly from those in mature hippocampus. The onset of HCN expression in interneurons of the hippocampus proper precedes that in the dentate gyrus, suggesting that HCN-mediated pacing activity may be generated in hippocampal interneurons prior to those in the hilus. Taken together, these findings indicate an age-dependent spatiotemporal evolution of specific HCN expression in distinct hippocampal cell populations, and suggest that these channels serve differing and evolving functions in the maturation of coordinated hippocampal activity. PMID:11682156

  19. The role of hyperpolarization-activated cationic current in spike-time precision and intrinsic resonance in cortical neurons in vitro.

    PubMed

    Gastrein, Philippe; Campanac, Emilie; Gasselin, Célia; Cudmore, Robert H; Bialowas, Andrzej; Carlier, Edmond; Fronzaroli-Molinieres, Laure; Ankri, Norbert; Debanne, Dominique

    2011-08-01

    Hyperpolarization-activated cyclic nucleotide modulated current (I(h)) sets resonance frequency within the θ-range (5–12 Hz) in pyramidal neurons. However, its precise contribution to the temporal fidelity of spike generation in response to stimulation of excitatory or inhibitory synapses remains unclear. In conditions where pharmacological blockade of I(h) does not affect synaptic transmission, we show that postsynaptic h-channels improve spike time precision in CA1 pyramidal neurons through two main mechanisms. I(h) enhances precision of excitatory postsynaptic potential (EPSP)--spike coupling because I(h) reduces peak EPSP duration. I(h) improves the precision of rebound spiking following inhibitory postsynaptic potentials (IPSPs) in CA1 pyramidal neurons and sets pacemaker activity in stratum oriens interneurons because I(h) accelerates the decay of both IPSPs and after-hyperpolarizing potentials (AHPs). The contribution of h-channels to intrinsic resonance and EPSP waveform was comparatively much smaller in CA3 pyramidal neurons. Our results indicate that the elementary mechanisms by which postsynaptic h-channels control fidelity of spike timing at the scale of individual neurons may account for the decreased theta-activity observed in hippocampal and neocortical networks when h-channel activity is pharmacologically reduced.

  20. The conduction system and expressions of hyperpolarization-activated cyclic nucleotide-gated cation channel 4 and connexin43 expressions in the hearts of fetal day 13 mice.

    PubMed

    Wen, Y; Li, B

    2017-01-01

    We investigated the development of the sinus node of the heart conduction system by localizing hyperpolarization-activated cyclic nucleotide-gated cation channel 4 (HCN4) and connexin43 (Cx43) in the hearts of fetal day 13 mice. Horizontal serial sections of day 13 whole fetuses were stained by hematoxylin and eosin and immunofluorescence to identify myocardial cells that express HCN4, hyperpolarization-activated cyclic nucleotide-gated cation channel 2 (HCN2) and Cx43. Expression levels of HCN4 and Cx43 were determined by quantitative RT-PCR in both fetal day 13 and adult mice. We found that both Cx43 and HCN4 expressions were located on the cell membranes in the hearts of fetal day 13 mice, but Cx43 was distributed throughout the myocardial cells. HCN4 expression was concentrated mainly in the left dorsal epicardium of the right atrium where Cx43 expression was low or absent. Quantitative RT-PCR demonstrated that HCN4 expression was significantly higher and HCN2 expression was significantly lower in fetal day 13 mice than in adults. We found no statistically significant difference in Cx43 expression between fetal day 13 mice and adults. HCN4 stained myocardial cells in the left dorsal epicardium of the right atrium are the origin of the sinus node and the remainder of the heart conduction system.

  1. Zn(2+) induces hyperpolarization by activation of a K(+) channel and increases intracellular Ca(2+) and pH in sea urchin spermatozoa.

    PubMed

    Beltrán, Carmen; Rodríguez-Miranda, Esmeralda; Granados-González, Gisela; García de De la Torre, Lucia; Nishigaki, Takuya; Darszon, Alberto

    2014-10-01

    Zinc (Zn(2+)) has been recently recognized as a crucial element for male gamete function in many species although its detailed mechanism of action is poorly understood. In sea urchin spermatozoa, Zn(2+) was reported as an essential trace ion for efficient sperm motility initiation and the acrosome reaction by modulating intracellular pH (pHi). In this study we found that submicromolar concentrations of free Zn(2+) change membrane potential (Em) and increase the concentration of intracellular Ca(2+) ([Ca(2+)]i) and cAMP in Lytechinus pictus sperm. Our results indicate that the Zn(2+) response in sperm of this species mainly involves an Em hyperpolarization caused by K(+) channel activation. The pharmacological profile of the Zn(2+)-induced hyperpolarization indicates that the cGMP-gated K(+) selective channel (tetraKCNG/CNGK), which is crucial for speract signaling, is likely a main target for Zn(2+). Considering that Zn(2+) also induces [Ca(2+)]i fluctuations, our observations suggest that Zn(2+) activates the signaling cascade of speract, except for an increase in cGMP, and facilitates sperm motility initiation upon spawning. These findings provide new insights about the role of Zn(2+) in male gamete function.

  2. Tetramerization Dynamics of C-terminal Domain Underlies Isoform-specific cAMP Gating in Hyperpolarization-activated Cyclic Nucleotide-gated Channels*

    PubMed Central

    Lolicato, Marco; Nardini, Marco; Gazzarrini, Sabrina; Möller, Stefan; Bertinetti, Daniela; Herberg, Friedrich W.; Bolognesi, Martino; Martin, Holger; Fasolini, Marina; Bertrand, Jay A.; Arrigoni, Cristina; Thiel, Gerhard; Moroni, Anna

    2011-01-01

    Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are dually activated by hyperpolarization and binding of cAMP to their cyclic nucleotide binding domain (CNBD). HCN isoforms respond differently to cAMP; binding of cAMP shifts activation of HCN2 and HCN4 by 17 mV but shifts that of HCN1 by only 2–4 mV. To explain the peculiarity of HCN1, we solved the crystal structures and performed a biochemical-biophysical characterization of the C-terminal domain (C-linker plus CNBD) of the three isoforms. Our main finding is that tetramerization of the C-terminal domain of HCN1 occurs at basal cAMP concentrations, whereas those of HCN2 and HCN4 require cAMP saturating levels. Therefore, HCN1 responds less markedly than HCN2 and HCN4 to cAMP increase because its CNBD is already partly tetrameric. This is confirmed by voltage clamp experiments showing that the right-shifted position of V½ in HCN1 is correlated with its propensity to tetramerize in vitro. These data underscore that ligand-induced CNBD tetramerization removes tonic inhibition from the pore of HCN channels. PMID:22006928

  3. Combined measurement of pulmonary inert gas washout and regional ventilation heterogeneity by MR of a single dose of hyperpolarized 3He.

    PubMed

    Deppe, Martin H; Parra-Robles, Juan; Ajraoui, Salma; Wild, Jim M

    2011-04-01

    Washout of inert gases is a measure of pulmonary function well-known in lung physiology. This work presents a method combining inert gas washout and spatially resolved imaging using hyperpolarized (3) He, thus providing complementary information on lung function and physiology. The nuclear magnetic resonance signal of intrapulmonary hyperpolarized (3) He is used to track the total amount of gas present within the lungs during multiple-breath washout via tidal breathing. Before the washout phase, 3D ventilation images are acquired using (3) He magnetic resonance imaging from the same dose of inhaled gas. The measured washout signal is corrected for T(1) relaxation and radiofrequency depletion, converting it into a quantity proportional to the apparent amount of gas within the lungs. The use of a pneumotachograph for acquisition of breathing volumes during washout, together with lung volumes derived from the magnetic resonance imaging data, permits assessment of the washout curves against physiological model predictions for healthy lungs. The shape of the resulting washout curves obtained from healthy volunteers matches the predictions, demonstrating the utility of the technique for the quantitative assessment of lung function. The proposed method can be readily integrated with a standard breath-hold (3) He ventilation imaging sequence, thus providing additional information from a single dose of gas.

  4. Real-time tracking of dissociation of hyperpolarized 89Y-DTPA: a model for degradation of open-chain Gd3+ MRI contrast agents

    NASA Astrophysics Data System (ADS)

    Ferguson, Sarah; Niedbalski, Peter; Parish, Christopher; Kiswandhi, Andhika; Kovacs, Zoltan; Lumata, Lloyd

    Gadolinium (Gd) complexes are widely used relaxation-based clinical contrast agents in magnetic resonance imaging (MRI). Gd-based MRI contrast agents with open-chain ligand such as Gd-DTPA, commercially known as magnevist, are less stable compared to Gd complexes with macrocyclic ligands such as GdDOTA (Dotarem). The dissociation of Gd-DPTA into Gd ion and DTPA ligand under certain biological conditions such as high zinc levels can potentially cause kidney damage. Since Gd is paramagnetic, direct NMR detection of the Gd-DTPA dissociation is quite challenging due to ultra-short relaxation times. In this work, we have investigated Y-DTPA as a model for Gd-DPTA dissociation under high zinc content solutions. Using dissolution dynamic nuclear polarization (DNP), the 89Y NMR signal is amplified by several thousand-fold. Due to the the relatively long T1 relaxation time of 89Y which translates to hyperpolarization lifetime of several minutes, the dissociation of Y-DTPA can be tracked in real-time by hyperpolarized 89Y NMR spectroscopy. Dissociation kinetic rates and implications on the degradation of open-chain Gd3+ MRI contrast agents will be discussed. This work was supported by the U.S. Department of Defense Award Number W81XWH-14-1-0048 and by the Robert A. Welch Foundation research Grant Number AT-1877.

  5. Diffusion-mediated 129Xe gas depolarization in magnetic field gradients during continuous-flow optical pumping

    NASA Astrophysics Data System (ADS)

    Burant, Alex; Branca, Rosa Tamara

    2016-12-01

    The production of large volumes of highly polarized noble gases like helium and xenon is vital to applications of magnetic resonance imaging and spectroscopy with hyperpolarized (HP) gas in humans. In the past ten years, 129Xe has become the gas of choice due to its lower cost, higher availability, relatively high tissue solubility, and wide range of chemical shift values. Though near unity levels of xenon polarization have been achieved in-cell using stopped-flow Spin Exchange Optical Pumping (SEOP), these levels are currently unmatched by continuous-flow SEOP methods. Among the various mechanisms that cause xenon relaxation, such as persistent and transient xenon dimers, wall collisions, and interactions with oxygen, relaxation due to diffusion in magnetic field gradients, caused by rapidly changing magnetic field strength and direction, is often ignored. However, during continuous-flow SEOP production, magnetic field gradients may not have a negligible contribution, especially considering that this methodology requires the combined use of magnets with very different characteristics (low field for spin exchange optical pumping and high field for the reduction of xenon depolarization in the solid state during the freeze out phase) that, when placed together, inevitably create magnetic field gradients along the gas-flow-path. Here, a combination of finite element analysis and Monte Carlo simulations is used to determine the effect of such magnetic field gradients on xenon gas polarization with applications to a specific, continuous-flow hyperpolarization system.

  6. Diffusion-mediated (129)Xe gas depolarization in magnetic field gradients during continuous-flow optical pumping.

    PubMed

    Burant, Alex; Branca, Rosa Tamara

    2016-12-01

    The production of large volumes of highly polarized noble gases like helium and xenon is vital to applications of magnetic resonance imaging and spectroscopy with hyperpolarized (HP) gas in humans. In the past ten years, (129)Xe has become the gas of choice due to its lower cost, higher availability, relatively high tissue solubility, and wide range of chemical shift values. Though near unity levels of xenon polarization have been achieved in-cell using stopped-flow Spin Exchange Optical Pumping (SEOP), these levels are currently unmatched by continuous-flow SEOP methods. Among the various mechanisms that cause xenon relaxation, such as persistent and transient xenon dimers, wall collisions, and interactions with oxygen, relaxation due to diffusion in magnetic field gradients, caused by rapidly changing magnetic field strength and direction, is often ignored. However, during continuous-flow SEOP production, magnetic field gradients may not have a negligible contribution, especially considering that this methodology requires the combined use of magnets with very different characteristics (low field for spin exchange optical pumping and high field for the reduction of xenon depolarization in the solid state during the freeze out phase) that, when placed together, inevitably create magnetic field gradients along the gas-flow-path. Here, a combination of finite element analysis and Monte Carlo simulations is used to determine the effect of such magnetic field gradients on xenon gas polarization with applications to a specific, continuous-flow hyperpolarization system.

  7. Adult and Continuing Education: Continuities and Discontinuities

    ERIC Educational Resources Information Center

    Tennant, Mark

    2005-01-01

    This paper presents a comprehensive review of the five-volume set, "Adult and Continuing Education." This publication is a monumental and diverse collection of material related directly or indirectly to the broad theme of adult and continuing education. The collection captures a great deal of the thought which has informed the…

  8. Detection of radiation induced lung injury in rats using dynamic hyperpolarized {sup 129}Xe magnetic resonance spectroscopy

    SciTech Connect

    Fox, Matthew S.; Ouriadov, Alexei; Hegarty, Elaine; Thind, Kundan; Wong, Eugene; Hope, Andrew; Santyr, Giles E.

    2014-07-15

    Purpose: Radiation induced lung injury (RILI) is a common side effect for patients undergoing thoracic radiation therapy (RT). RILI can lead to temporary or permanent loss of lung function and in extreme cases, death. Combining functional lung imaging information with conventional radiation treatment plans may lead to more desirable treatment plans that reduce lung toxicity and improve the quality of life for lung cancer survivors. Magnetic Resonance Imaging of the lung following inhalation of hyperpolarized{sup 129}Xe may provide a useful nonionizing approach for probing changes in lung function and structure associated with RILI before, during, or after RT (early and late time-points). Methods: In this study, dynamic{sup 129}Xe MR spectroscopy was used to measure whole-lung gas transfer time constants for lung tissue and red blood cells (RBC), respectively (T{sub Tr-tissue} and T{sub Tr-RBC}) in groups of rats at two weeks and six weeks following 14 Gy whole-lung exposure to radiation from a {sup 60}Co source. A separate group of six healthy age-matched rats served as a control group. Results: T{sub Tr-tissue} values at two weeks post-irradiation (51.6 ± 6.8 ms) were found to be significantly elevated (p < 0.05) with respect to the healthy control group (37.2 ± 4.8 ms). T{sub Tr-RBC} did not show any significant changes between groups. T{sub Tr-tissue} was strongly correlated with T{sub Tr-RBC} in the control group (r = 0.9601 p < 0.05) and uncorrelated in the irradiated groups. Measurements of arterial partial pressure of oxygen obtained by arterial blood sampling were found to be significantly decreased (p < 0.05) in the two-week group (54.2 ± 12.3 mm Hg) compared to those from a representative control group (85.0 ± 10.0 mm Hg). Histology of a separate group of similarly irradiated animals confirmed the presence of inflammation due to radiation exposure with alveolar wall thicknesses that were significantly different (p < 0.05). At six weeks post

  9. Protective Effect and Mechanism of Total Flavones from Rhododendron simsii Planch on Endothelium-Dependent Dilatation and Hyperpolarization in Cerebral Ischemia-Reperfusion and Correlation to Hydrogen Sulphide Release in Rats

    PubMed Central

    Han, Jun; He, Guo-Wei; Chen, Zhi-Wu

    2014-01-01

    We for the first time investigated the effect and mechanism of the total flavones of Rhododendron simsii Planch (TFR), a widely-used Chinese herb for a thousand years, on vasodilatation and hyperpolarization in middle cerebral artery (MCA) of rats subject to global cerebral ischemia-reperfusion (CIR). TFR (11~2700 mg/L) evoked dose-dependent vasodilation and hyperpolarization in MCA of both sham and CIR that were partially inhibited by 30 μM N-nitro-L-arginine-methyl-ester and 10 μM indomethacin and further attenuated by endogenous H2S synthese-CSE inhibitor PPG (100 μM) or Ca2+-activated potassium channel (Kca) inhibitor TEA (1 mM). In whole-cell patch clamp recording, TFR remarkably enhanced the outward current that was inhibited by TEA. CIR increased CSE mRNA expression and the contents of H2S that were further increased by TFR. We conclude that, in MCA of CIR rats, TFR induces non-NO and non-PGI2-mediated effects of vasodilatation and hyperpolarization involving Kca and increases CSE mRNA expression level in endothelial cells and H2S content in the cerebrum. These findings suggest that the response induced by TFR is potentially related to endothelium-derived hyperpolarizing factor mediated by the endogenous H2S and promote the use of TFR in protection of brain from ischemia-reperfusion injury. PMID:25050128

  10. Difference between Extra- and Intracellular T1 Values of Carboxylic Acids Affects the Quantitative Analysis of Cellular Kinetics by Hyperpolarized NMR.

    PubMed

    Karlsson, Magnus; Jensen, Pernille Rose; Ardenkjaer-Larsen, Jan Henrik; Lerche, Mathilde H

    2016-10-17

    Incomplete knowledge of the longitudinal relaxation time constant (T1 ) leads to incorrect assumptions in quantitative kinetic models of cellular systems, studied by hyperpolarized real-time NMR. Using an assay that measures the intracellular signal of small carboxylic acids in living cells, the intracellular T1 of the carboxylic acid moiety of acetate, keto-isocaproate, pyruvate, and butyrate was determined. The intracellular T1 is shown to be up to four-fold shorter than the extracellular T1 . Such a large difference in T1 values between the inside and the outside of the cell has significant influence on the quantification of intracellular metabolic activity. It is expected that the significantly shorter T1 value of the carboxylic moieties inside cells is a result of macromolecular crowding. An artificial cytosol has been prepared and applied to predict the T1 of other carboxylic acids. We demonstrate the value of this prediction tool.

  11. Multi-channel metabolic imaging, with SENSE reconstruction, of hyperpolarized [1- 13C] pyruvate in a live rat at 3.0 tesla on a clinical MR scanner

    NASA Astrophysics Data System (ADS)

    Tropp, James; Lupo, Janine M.; Chen, Albert; Calderon, Paul; McCune, Don; Grafendorfer, Thomas; Ozturk-Isik, Esin; Larson, Peder E. Z.; Hu, Simon; Yen, Yi-Fen; Robb, Fraser; Bok, Robert; Schulte, Rolf; Xu, Duan; Hurd, Ralph; Vigneron, Daniel; Nelson, Sarah

    2011-01-01

    We report metabolic images of 13C, following injection of a bolus of hyperpolarized [1-13C] pyruvate in a live rat. The data were acquired on a clinical scanner, using custom coils for volume transmission and array reception. Proton blocking of all carbon resonators enabled proton anatomic imaging with the system body coil, to allow for registration of anatomic and metabolic images, for which good correlation was achieved, with some anatomic features (kidney and heart) clearly visible in a carbon image, without reference to the corresponding proton image. Parallel imaging with sensitivity encoding was used to increase the spatial resolution in the SI direction of the rat. The signal to noise ratio in was in some instances unexpectedly high in the parallel images; variability of the polarization among different trials, plus partial volume effects, are noted as a possible cause of this.

  12. Cellular adaptation to anthrax lethal toxin-induced mitochondrial cholesterol enrichment, hyperpolarization, and reactive oxygen species generation through downregulating MLN64 in macrophages.

    PubMed

    Ha, Soon-Duck; Park, Sangwook; Han, Chae Young; Nguyen, Marilyn L; Kim, Sung Ouk

    2012-12-01

    Cellular adaptation to different stresses related to survival and function has been demonstrated in several cell types. Anthrax lethal toxin (LeTx) induces rapid cell death, termed "pyroptosis," by activating NLRP1b/caspase-1 in murine macrophages. We and others (S. D. Ha et al., J. Biol. Chem. 282:26275-26283, 2007; I. I. Salles et al., Proc. Natl. Acad. Sci. U. S. A. 100:12426 -12431, 2003) have shown that RAW264.7 cells preexposed to sublethal doses of LeTx become resistant to subsequent high cytolytic doses of LeTx, termed toxin-induced resistance (TIR). To date, the cellular mechanisms of pyroptosis and TIR are largely unknown. We found that LeTx caused NLRP1b/caspase-1-dependent mitochondrial dysfunction, including hyperpolarization and generation of reactive oxygen species, which was distinct from that induced by stimuli such as NLRP3-activating ATP. In TIR cells, these mitochondrial events were not detected, although caspase-1 was activated, in response to LeTx. We identified that downregulation of the late endosomal cholesterol-transferring protein MLN64 in TIR cells was involved in TIR. The downregulation of MLN64 in TIR cells was at least in part due to DNA methyltransferase 1-mediated DNA methylation. In wild-type RAW264.7 cells and primary bone marrow-derived macrophages, LeTx caused NLRP1b/caspase-1-dependent mitochondrial translocation of MLN64, resulting in cholesterol enrichment, membrane hyperpolarization, reactive oxygen species (ROS) generation, and depletion of free glutathione (GSH). This study demonstrates for the first time that MLN64 plays a key role in LeTx/caspase-1-induced mitochondrial dysfunction.

  13. Small-fiber neuropathy Nav1.8 mutation shifts activation to hyperpolarized potentials and increases excitability of dorsal root ganglion neurons.

    PubMed

    Huang, Jianying; Yang, Yang; Zhao, Peng; Gerrits, Monique M; Hoeijmakers, Janneke G J; Bekelaar, Kim; Merkies, Ingemar S J; Faber, Catharina G; Dib-Hajj, Sulayman D; Waxman, Stephen G

    2013-08-28

    Idiopathic small-fiber neuropathy (I-SFN), clinically characterized by burning pain in distal extremities and autonomic dysfunction, is a disorder of small-caliber nerve fibers of unknown etiology with limited treatment options. Functional variants of voltage-gated sodium channel Nav1.7, encoded by SCN9A, have been identified in approximately one-third of I-SFN patients. These variants render dorsal root ganglion (DRG) neurons hyperexcitable. Sodium channel Nav1.8, encoded by SCN10A, is preferentially expressed in small-diameter DRG neurons, and produces most of the current underlying the upstroke of action potentials in these neurons. We previously demonstrated two functional variants of Nav1.8 that either enhance ramp current or shift activation in a hyperpolarizing direction, and render DRG neurons hyperexcitable, in I-SFN patients with no mutations of SCN9A. We have now evaluated additional I-SFN patients with no mutations in SCN9A, and report a novel I-SFN-related Nav1.8 mutation I1706V in a patient with painful I-SFN. Whole-cell voltage-clamp recordings in small DRG neurons demonstrate that the mutation hyperpolarizes activation and the response to slow ramp depolarizations. However, it decreases fractional channels resistant to fast inactivation and reduces persistent currents. Current-clamp studies reveal that mutant channels decrease current threshold and increase the firing frequency of evoked action potentials within small DRG neurons. These observations suggest that the effects of this mutation on activation and ramp current are dominant over the reduced persistent current, and show that these pro-excitatory gating changes confer hyperexcitability on peripheral sensory neurons, which may contribute to pain in this individual with I-SFN.

  14. Immunolocalization of hyperpolarization-activated cationic HCN1 and HCN3 channels in the rat nephron: regulation of HCN3 by potassium diets.

    PubMed

    López-González, Zinaeli; Ayala-Aguilera, Cosete; Martinez-Morales, Flavio; Galicia-Cruz, Othir; Salvador-Hernández, Carolina; Pedraza-Chaverri, José; Medeiros, Mara; Hernández, Ana Maria; Escobar, Laura I

    2016-01-01

    Hyperpolarization-activated cationic and cyclic nucleotide-gated channels (HCN) comprise four homologous subunits (HCN1-HCN4). HCN channels are found in excitable and non-excitable tissues in mammals. We have previously shown that HCN2 may transport ammonium (NH4 (+)), besides sodium (Na(+)), in the rat distal nephron. In the present work, we identified HCN1 and HCN3 in the proximal tubule (PT) and HCN3 in the thick ascending limb of Henle (TALH) of the rat kidney. Immunoblot assays detected HCN1 (130 kDa) and HCN3 (90 KDa) and their truncated proteins C-terminal HCN1 (93 KDa) and N-terminal HCN3 (65 KDa) in enriched plasma membranes from cortex (CX) and outer medulla (OM), as well as in brush-border membrane vesicles. Immunofluorescence assays confirmed apical localization of HCN1 and HCN3 in the PT. HCN3 was also found at the basolateral membrane of TALH. We evaluated chronic changes in mineral dietary on HCN3 protein abundance. Animals were fed with three different diets: sodium-deficient (SD) diet, potassium-deficient (KD) diet, and high-potassium (HK) diet. Up-regulation of HCN3 was observed in OM by KD and in CX and OM by HK; the opposite effect occurred with the N-terminal truncated HCN3 in CX (KD) and OM (HK). SD diet did not produce any change. Since HCN channels activate with membrane hyperpolarization, our results suggest that HCN channels may play a role in the Na(+)-K(+)-ATPase activity, contributing to Na(+), K(+), and acid-base homeostasis in the rat kidney.

  15. Development and application of methods to quantify spatial and temporal hyperpolarized 3He MRI ventilation dynamics: preliminary results in chronic obstructive pulmonary disease

    NASA Astrophysics Data System (ADS)

    Kirby, Miranda; Wheatley, Andrew; McCormack, David G.; Parraga, Grace

    2010-03-01

    Hyperpolarized helium-3 (3He) magnetic resonance imaging (MRI) has emerged as a non-invasive research method for quantifying lung structural and functional changes, enabling direct visualization in vivo at high spatial and temporal resolution. Here we described the development of methods for quantifying ventilation dynamics in response to salbutamol in Chronic Obstructive Pulmonary Disease (COPD). Whole body 3.0 Tesla Excite 12.0 MRI system was used to obtain multi-slice coronal images acquired immediately after subjects inhaled hyperpolarized 3He gas. Ventilated volume (VV), ventilation defect volume (VDV) and thoracic cavity volume (TCV) were recorded following segmentation of 3He and 1H images respectively, and used to calculate percent ventilated volume (PVV) and ventilation defect percent (VDP). Manual segmentation and Otsu thresholding were significantly correlated for VV (r=.82, p=.001), VDV (r=.87 p=.0002), PVV (r=.85, p=.0005), and VDP (r=.85, p=.0005). The level of agreement between these segmentation methods was also evaluated using Bland-Altman analysis and this showed that manual segmentation was consistently higher for VV (Mean=.22 L, SD=.05) and consistently lower for VDV (Mean=-.13, SD=.05) measurements than Otsu thresholding. To automate the quantification of newly ventilated pixels (NVp) post-bronchodilator, we used translation, rotation, and scaling transformations to register pre-and post-salbutamol images. There was a significant correlation between NVp and VDV (r=-.94 p=.005) and between percent newly ventilated pixels (PNVp) and VDP (r=- .89, p=.02), but not for VV or PVV. Evaluation of 3He MRI ventilation dynamics using Otsu thresholding and landmark-based image registration provides a way to regionally quantify functional changes in COPD subjects after treatment with beta-agonist bronchodilators, a common COPD and asthma therapy.

  16. N-methyl-D-aspartate, hyperpolarization-activated cation current (Ih) and gamma-aminobutyric acid conductances govern the risk of epileptogenesis following febrile seizures in rat hippocampus.

    PubMed

    Ouardouz, Mohamed; Lema, Pablo; Awad, Patricia N; Di Cristo, Graziella; Carmant, Lionel

    2010-04-01

    Febrile seizures are the most common types of seizure in children, and are generally considered to be benign. However, febrile seizures in children with dysgenesis have been associated with the development of temporal lobe epilepsy. We have previously shown in a rat model of dysgenesis (cortical freeze lesion) and hyperthermia-induced seizures that 86% of these animals developed recurrent seizures in adulthood. The cellular changes underlying the increased risk of epileptogenesis in this model are not known. Using whole cell patch-clamp recordings from CA1 hippocampal pyramidal cells, we found a more pronounced increase in excitability in rats with both hyperthermic seizures and dysgenesis than in rats with hyperthermic seizures alone or dysgenesis alone. The change was found to be secondary to an increase in N-methyl-D-aspartate (NMDA) receptor-mediated excitatory postsynaptic currents (EPSCs). Inversely, hyperpolarization-activated cation current was more pronounced in naïve rats with hyperthermic seizures than in rats with dysgenesis and hyperthermic seizures or with dysgenesis alone. The increase in GABAA-mediated inhibition observed was comparable in rats with or without dysgenesis after hyperthermic seizures, whereas no changes were observed in rats with dysgenesis alone. Our work indicates that in this two-hit model, changes in NMDA receptor-mediated EPSCs may facilitate epileptogenesis following febrile seizures. Changes in the hyperpolarization-activated cation currents may represent a protective reaction and act by damping the NMDA receptor-mediated hyperexcitability, rather than converting inhibition into excitation. These findings provide a new hypothesis of cellular changes following hyperthermic seizures in predisposed individuals, and may help in the design of therapeutic strategies to prevent epileptogenesis following prolonged febrile seizures.

  17. High resolution spectroscopy and chemical shift imaging of hyperpolarized 129Xe dissolved in the human brain in vivo at 1.5 tesla

    PubMed Central

    Rao, Madhwesha; Stewart, Neil J.; Norquay, Graham; Griffiths, Paul D.

    2016-01-01

    Purpose Upon inhalation, xenon diffuses into the bloodstream and is transported to the brain, where it dissolves in various compartments of the brain. Although up to five chemically distinct peaks have been previously observed in 129Xe rat head spectra, to date only three peaks have been reported in the human head. This study demonstrates high resolution spectroscopy and chemical shift imaging (CSI) of 129Xe dissolved in the human head at 1.5 Tesla. Methods A 129Xe radiofrequency coil was built in‐house and 129Xe gas was polarized using spin‐exchange optical pumping. Following the inhalation of 129Xe gas, NMR spectroscopy was performed with spectral resolution of 0.033 ppm. Two‐dimensional CSI in all three anatomical planes was performed with spectral resolution of 2.1 ppm and voxel size 20 mm × 20 mm. Results Spectra of hyperpolarized 129Xe dissolved in the human head showed five distinct peaks at 188 ppm, 192 ppm, 196 ppm, 200 ppm, and 217 ppm. Assignment of these peaks was consistent with earlier studies. Conclusion High resolution spectroscopy and CSI of hyperpolarized 129Xe dissolved in the human head has been demonstrated. For the first time, five distinct NMR peaks have been observed in 129Xe spectra from the human head in vivo. Magn Reson Med 75:2227–2234, 2016. © 2016 The Authors Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. PMID:27080441

  18. NO Hyperpolarizes Pulmonary Artery Smooth Muscle Cells and Decreases the Intracellular Ca2+ Concentration by Activating Voltage-Gated K+ Channels

    NASA Astrophysics Data System (ADS)

    Yuan, Xiao-Jian; Tod, Mary L.; Rubin, Lewis J.; Blaustein, Mordecai P.

    1996-09-01

    NO causes pulmonary vasodilation in patients with pulmonary hypertension. In pulmonary arterial smooth muscle cells, the activity of voltage-gated K+ (KV) channels controls resting membrane potential. In turn, membrane potential is an important regulator of the intracellular free calcium concentration ([Ca2+]i) and pulmonary vascular tone. We used patch clamp methods to determine whether the NO-induced pulmonary vasodilation is mediated by activation of KV channels. Quantitative fluorescence microscopy was employed to test the effect of NO on the depolarization-induced rise in [Ca2+]i. Blockade of KV channels by 4-aminopyridine (5 mM) depolarized pulmonary artery myocytes to threshold for initiation of Ca2+ action potentials, and thereby increased [Ca2+]i. NO (≈ 3 μ M) and the NO-generating compound sodium nitroprusside (5-10 μ M) opened KV channels in rat pulmonary artery smooth muscle cells. The enhanced K+ currents then hyperpolarized the cells, and blocked Ca2+-dependent action potentials, thereby preventing the evoked increases in [Ca2+]i. Nitroprusside also increased the probability of KV channel opening in excised, outside-out membrane patches. This raises the possibility that NO may act either directly on the channel protein or on a closely associated molecule rather than via soluble guanylate cyclase. In isolated pulmonary arteries, 4-aminopyridine significantly inhibited NO-induced relaxation. We conclude that NO promotes the opening of KV channels in pulmonary arterial smooth muscle cells. The resulting membrane hyperpolarization, which lowers [Ca2+]i, is apparently one of the mechanisms by which NO induces pulmonary vasodilation.

  19. Continuous-flow electrophoretic separator for biologicals

    NASA Technical Reports Server (NTRS)

    Mccreight, L. R.; Griffin, R. N.; Locker, R. J.

    1976-01-01

    In the near absence of gravity, a continuous-flow type of electrophoretic separator can be operated with a much thicker separation chamber than is possible under 1 g conditions. This should permit either better resolution or shorter separation time per unit of sample. An apparatus to perform experiments on sounding rockets is under development and will be described. The electrophoresis cell is 5 mm thick by 5 cm wide with 10 cm long electrodes. It is supplied with buffer, sample, and coolant at about 4 C through the use of a passive refrigerant system. UV sample detection and provision for recovery and cold storage of up to 50 sample fractions are now being added to the basic unit. A wide range of operating conditions are electronically programmable into the unit, even up to a short time before flight, and a further range of some parameters can be achieved by exchanging power supplies and by changing gears in the motor drive units of the pump. The preliminary results of some separation studies on various biological products using a commercially available electrophoretic separator are also presented.

  20. Programmes in Continuing Education

    ERIC Educational Resources Information Center

    Shah, L. R.

    1976-01-01

    The various types and forms of credit and non-credit university continuing education programmes are described in these extracts from a paper presented at the Hyderabad conference on university continuing education. (ABM)

  1. Uniform Continuity of POVMs

    NASA Astrophysics Data System (ADS)

    Beneduci, Roberto

    2014-10-01

    Recently a characterization of uniformly continuous POVMs and a necessary condition for a uniformly continuous POVM F to have the norm-1 property have been provided. Moreover it was proved that in the commutative case, uniform continuity corresponds to the existence of a Feller Markov kernel. We apply such results to the analysis of some relevant physical examples; i.e., the phase space localization observables, the unsharp phase observable and the unsharp number observable of which we study the uniform continuity, the norm-1 property and the existence of a Feller Markov kernel.

  2. A system for accurate and automated injection of hyperpolarized substrate with minimal dead time and scalable volumes over a large range.

    PubMed

    Reynolds, Steven; Bucur, Adriana; Port, Michael; Alizadeh, Tooba; Kazan, Samira M; Tozer, Gillian M; Paley, Martyn N J

    2014-02-01

    Over recent years hyperpolarization by dissolution dynamic nuclear polarization has become an established technique for studying metabolism in vivo in animal models. Temporal signal plots obtained from the injected metabolite and daughter products, e.g. pyruvate and lactate, can be fitted to compartmental models to estimate kinetic rate constants. Modeling and physiological parameter estimation can be made more robust by consistent and reproducible injections through automation. An injection system previously developed by us was limited in the injectable volume to between 0.6 and 2.4ml and injection was delayed due to a required syringe filling step. An improved MR-compatible injector system has been developed that measures the pH of injected substrate, uses flow control to reduce dead volume within the injection cannula and can be operated over a larger volume range. The delay time to injection has been minimized by removing the syringe filling step by use of a peristaltic pump. For 100μl to 10.000ml, the volume range typically used for mice to rabbits, the average delivered volume was 97.8% of the demand volume. The standard deviation of delivered volumes was 7μl for 100μl and 20μl for 10.000ml demand volumes (mean S.D. was 9 ul in this range). In three repeat injections through a fixed 0.96mm O.D. tube the coefficient of variation for the area under the curve was 2%. For in vivo injections of hyperpolarized pyruvate in tumor-bearing rats, signal was first detected in the input femoral vein cannula at 3-4s post-injection trigger signal and at 9-12s in tumor tissue. The pH of the injected pyruvate was 7.1±0.3 (mean±S.D., n=10). For small injection volumes, e.g. less than 100μl, the internal diameter of the tubing contained within the peristaltic pump could be reduced to improve accuracy. Larger injection volumes are limited only by the size of the receiving vessel connected to the pump.

  3. A system for accurate and automated injection of hyperpolarized substrate with minimal dead time and scalable volumes over a large range☆

    PubMed Central

    Reynolds, Steven; Bucur, Adriana; Port, Michael; Alizadeh, Tooba; Kazan, Samira M.; Tozer, Gillian M.; Paley, Martyn N.J.

    2014-01-01

    Over recent years hyperpolarization by dissolution dynamic nuclear polarization has become an established technique for studying metabolism in vivo in animal models. Temporal signal plots obtained from the injected metabolite and daughter products, e.g. pyruvate and lactate, can be fitted to compartmental models to estimate kinetic rate constants. Modeling and physiological parameter estimation can be made more robust by consistent and reproducible injections through automation. An injection system previously developed by us was limited in the injectable volume to between 0.6 and 2.4 ml and injection was delayed due to a required syringe filling step. An improved MR-compatible injector system has been developed that measures the pH of injected substrate, uses flow control to reduce dead volume within the injection cannula and can be operated over a larger volume range. The delay time to injection has been minimized by removing the syringe filling step by use of a peristaltic pump. For 100 μl to 10.000 ml, the volume range typically used for mice to rabbits, the average delivered volume was 97.8% of the demand volume. The standard deviation of delivered volumes was 7 μl for 100 μl and 20 μl for 10.000 ml demand volumes (mean S.D. was 9 ul in this range). In three repeat injections through a fixed 0.96 mm O.D. tube the coefficient of variation for the area under the curve was 2%. For in vivo injections of hyperpolarized pyruvate in tumor-bearing rats, signal was first detected in the input femoral vein cannula at 3–4 s post-injection trigger signal and at 9–12 s in tumor tissue. The pH of the injected pyruvate was 7.1 ± 0.3 (mean ± S.D., n = 10). For small injection volumes, e.g. less than 100 μl, the internal diameter of the tubing contained within the peristaltic pump could be reduced to improve accuracy. Larger injection volumes are limited only by the size of the receiving vessel connected to the pump. PMID:24355621

  4. A system for accurate and automated injection of hyperpolarized substrate with minimal dead time and scalable volumes over a large range

    NASA Astrophysics Data System (ADS)

    Reynolds, Steven; Bucur, Adriana; Port, Michael; Alizadeh, Tooba; Kazan, Samira M.; Tozer, Gillian M.; Paley, Martyn N. J.

    2014-02-01

    Over recent years hyperpolarization by dissolution dynamic nuclear polarization has become an established technique for studying metabolism in vivo in animal models. Temporal signal plots obtained from the injected metabolite and daughter products, e.g. pyruvate and lactate, can be fitted to compartmental models to estimate kinetic rate constants. Modeling and physiological parameter estimation can be made more robust by consistent and reproducible injections through automation. An injection system previously developed by us was limited in the injectable volume to between 0.6 and 2.4 ml and injection was delayed due to a required syringe filling step. An improved MR-compatible injector system has been developed that measures the pH of injected substrate, uses flow control to reduce dead volume within the injection cannula and can be operated over a larger volume range. The delay time to injection has been minimized by removing the syringe filling step by use of a peristaltic pump. For 100 μl to 10.000 ml, the volume range typically used for mice to rabbits, the average delivered volume was 97.8% of the demand volume. The standard deviation of delivered volumes was 7 μl for 100 μl and 20 μl for 10.000 ml demand volumes (mean S.D. was 9 ul in this range). In three repeat injections through a fixed 0.96 mm O.D. tube the coefficient of variation for the area under the curve was 2%. For in vivo injections of hyperpolarized pyruvate in tumor-bearing rats, signal was first detected in the input femoral vein cannula at 3-4 s post-injection trigger signal and at 9-12 s in tumor tissue. The pH of the injected pyruvate was 7.1 ± 0.3 (mean ± S.D., n = 10). For small injection volumes, e.g. less than 100 μl, the internal diameter of the tubing contained within the peristaltic pump could be reduced to improve accuracy. Larger injection volumes are limited only by the size of the receiving vessel connected to the pump.

  5. The Tactile Continuity Illusion

    ERIC Educational Resources Information Center

    Kitagawa, Norimichi; Igarashi, Yuka; Kashino, Makio

    2009-01-01

    We can perceive the continuity of an object or event by integrating spatially/temporally discrete sensory inputs. The mechanism underlying this perception of continuity has intrigued many researchers and has been well documented in both the visual and auditory modalities. The present study shows for the first time to our knowledge that an illusion…

  6. Reinventing Continuing Higher Education

    ERIC Educational Resources Information Center

    Walshok, Mary Lindenstein

    2012-01-01

    Re-inventing continuing higher education is about finding ways to be a more central player in a region's civic, cultural, and economic life as well as in the education of individuals for work and citizenship. Continuing higher education will require data gathering, analytical tools, convening authority, interpretive skills, new models of delivery,…

  7. Continuing Vocational Training (CVT).

    ERIC Educational Resources Information Center

    Drogosz-Zablocka, Elzbieta; Izycka, Halina; Trzeciak, Wlodzimierz

    Polish continuing education (CE) includes education, further education, and professional development in and out of school; in day, evening, or weekend courses; and distance education. The state, workplaces, grants, and foreign assistance provide financing. A variety of organizations cooperate to provide continuing education. High-risk groups…

  8. Residential Continuing Education.

    ERIC Educational Resources Information Center

    Houle, Cyril O.

    The theme of this discursive essay is residential continuing education: its definition, its development along somewhat different lines in Europe and in America, and its practice in university centers in the United States. Continuing education includes any learning or teaching program that is based on the assumptions that the learners have studied…

  9. Establishing a Continuous Repertoire.

    ERIC Educational Resources Information Center

    Meadowcroft, Pamela; Holland, James G.

    Investigators in this study looked for conditions that can rapidly establish continuous stimulus control of continuous response variations, or "response mapping." Unlike previous research in stimulus control, where a single stimulus comes to control a single response, 36 5-year-old children received errorless discrimination training at…

  10. Use of hyperpolarized [1-13C]pyruvate and [2-13C]pyruvate to probe the effects of the anticancer agent dichloroacetate on mitochondrial metabolism in vivo in the normal rat.

    PubMed

    Hu, Simon; Yoshihara, Hikari A I; Bok, Robert; Zhou, Jenny; Zhu, Minhua; Kurhanewicz, John; Vigneron, Daniel B

    2012-12-01

    Development of hyperpolarized technology utilizing dynamic nuclear polarization has enabled the measurement of (13)C metabolism in vivo at very high signal-to-noise ratio (SNR). In vivo mitochondrial metabolism can, in principle, be monitored with pyruvate, which is catalyzed to acetyl-CoA via pyruvate dehydrogenase (PDH). The purpose of this work was to determine whether the compound sodium dichloroacetate (DCA) could aid the study of mitochondrial metabolism with hyperpolarized pyruvate. DCA stimulates PDH by inhibiting its inhibitor, pyruvate dehydrogenase kinase. In this work, hyperpolarized [1-(13)C]pyruvate and [2-(13)C]pyruvate were used to probe mitochondrial metabolism in normal rats. Increased conversion to bicarbonate (+181±69%, P=.025) was measured when [1-(13)C]pyruvate was injected after DCA administration, and increased glutamate (+74±23%, P=.004), acetoacetate (+504±281%, P=.009) and acetylcarnitine (+377±157%, P=.003) were detected when [2-(13)C]pyruvate was used.

  11. Column continuous transition functions

    NASA Astrophysics Data System (ADS)

    Li, Yangrong

    2007-04-01

    A column continuous transition function is by definition a standard transition function P(t) whose every column is continuous for t[greater-or-equal, slanted]0 in the norm topology of bounded sequence space l[infinity]. We will prove that it has a stable q-matrix and that there exists a one-to-one relationship between column continuous transition functions and increasing integrated semigroups on l[infinity]. Using the theory of integrated semigroups, we give some necessary and sufficient conditions under which the minimal q-function is column continuous, in terms of its generator (of the Markov semigroup) as well as its q-matrix. Furthermore, we will construct all column continuous Q-functions for a conservative, single-exit and column bounded q-matrix Q. As applications, we find that many interesting continuous-time Markov chains (CTMCs), say Feller-Reuter-Riley processes, monotone processes, birth-death processes and branching processes, etc., have column continuity.

  12. Probing Porosity and Pore Interconnectivity in Self-Assembled TiO2–Graphene Hybrid Nanostructures Using Hyperpolarized 129Xe NMR

    SciTech Connect

    Wang, Li Qiong; Wang, Donghai; Liu, Jun; Exarhos, Gregory J.

    2012-01-12

    Hyperpolarized (HP) {sup 129}Xe NMR was used to probe the porosity and interconnectivity of pores in self-assembled hybrid TiO{sub 2}-graphene nanostructures. We have demonstrated that HP {sup 129}Xe NMR is a powerful technique in probing any changes in porosity and interconnectivity of the pores caused by addition of a small amount of functionalized graphene sheets (FGSs) (1% weight percent) into the network of mesoporous TiO{sub 2}. To obtain the information on the changes in porosity and interconnectivity of the pores caused by addition of a small amount of FGSs, a comparative study has been carried out by acquiring HP {sup 129}Xe NMR spectra under identical experimental conditions for both pure mesoporous TiO{sub 2} and hybrid TiO{sub 2}-FGSs. The HP {sup 129}Xe NMR results from our comparative study suggest that TiO{sub 2} and graphene are mixed uniformly on the nanoscale and the resulting hybrid nanostructure has better channel connectivity between different domains, enhancing the transport property for Li-insertion/extraction.

  13. EPR oxygen imaging and hyperpolarized 13C MRI of pyruvate metabolism as noninvasive biomarkers of tumor treatment response to a glycolysis inhibitor 3-bromopyruvate.

    PubMed

    Matsumoto, Shingo; Saito, Keita; Yasui, Hironobu; Morris, H Douglas; Munasinghe, Jeeva P; Lizak, Martin; Merkle, Hellmut; Ardenkjaer-Larsen, Jan Henrik; Choudhuri, Rajani; Devasahayam, Nallathamby; Subramanian, Sankaran; Koretsky, Alan P; Mitchell, James B; Krishna, Murali C

    2013-05-01

    The hypoxic nature of tumors results in treatment resistance and poor prognosis. To spare limited oxygen for more crucial pathways, hypoxic cancerous cells suppress mitochondrial oxidative phosphorylation and promote glycolysis for energy production. Thereby, inhibition of glycolysis has the potential to overcome treatment resistance of hypoxic tumors. Here, EPR imaging was used to evaluate oxygen dependent efficacy on hypoxia-sensitive drug. The small molecule 3-bromopyruvate blocks glycolysis pathway by inhibiting hypoxia inducible enzymes and enhanced cytotoxicity of 3-bromopyruvate under hypoxic conditions has been reported in vitro. However, the efficacy of 3-bromopyruvate was substantially attenuated in hypoxic tumor regions (pO2<10 mmHg) in vivo using squamous cell carcinoma (SCCVII)-bearing mouse model. Metabolic MRI studies using hyperpolarized 13C-labeled pyruvate showed that monocarboxylate transporter-1 is the major transporter for pyruvate and the analog 3-bromopyruvate in SCCVII tumor. The discrepant results between in vitro and in vivo data were attributed to biphasic oxygen dependent expression of monocarboxylate transporter-1 in vivo. Expression of monocarboxylate transporter-1 was enhanced in moderately hypoxic (8-15 mmHg) tumor regions but down regulated in severely hypoxic (<5 mmHg) tumor regions. These results emphasize the importance of noninvasive imaging biomarkers to confirm the action of hypoxia-activated drugs.

  14. Simultaneous investigation of cardiac pyruvate dehydrogenase flux, Krebs cycle metabolism and pH, using hyperpolarized [1,2-(13)C2]pyruvate in vivo.

    PubMed

    Chen, Albert P; Hurd, Ralph E; Schroeder, Marie A; Lau, Angus Z; Gu, Yi-ping; Lam, Wilfred W; Barry, Jennifer; Tropp, James; Cunningham, Charles H

    2012-02-01

    (13)C MR spectroscopy studies performed on hearts ex vivo and in vivo following perfusion of prepolarized [1-(13)C]pyruvate have shown that changes in pyruvate dehydrogenase (PDH) flux may be monitored non-invasively. However, to allow investigation of Krebs cycle metabolism, the (13)C label must be placed on the C2 position of pyruvate. Thus, the utilization of either C1 or C2 labeled prepolarized pyruvate as a tracer can only afford a partial view of cardiac pyruvate metabolism in health and disease. If the prepolarized pyruvate molecules were labeled at both C1 and C2 positions, then it would be possible to observe the downstream metabolites that were the results of both PDH flux ((13)CO(2) and H(13)CO(3)(-)) and Krebs cycle flux ([5-(13)C]glutamate) with a single dose of the agent. Cardiac pH could also be monitored in the same experiment, but adequate SNR of the (13)CO(2) resonance may be difficult to obtain in vivo. Using an interleaved selective RF pulse acquisition scheme to improve (13)CO(2) detection, the feasibility of using dual-labeled hyperpolarized [1,2-(13)C(2)]pyruvate as a substrate for dynamic cardiac metabolic MRS studies to allow simultaneous investigation of PDH flux, Krebs cycle flux and pH, was demonstrated in vivo.

  15. Probing S4 and S5 Segment Proximity in Mammalian Hyperpolarization Activated HCN Channels by Disulfide Bridging and Cd2+ Coordination

    PubMed Central

    Bell, Damian C.; Turbendian, Harma K.; Valley, Matthew T.; Zhou, Lei; Riley, John H.; Siegelbaum, Steven A.; Tibbs, Gareth R.

    2009-01-01

    We explored the structural basis of voltage sensing in HCN1 hyperpolarization-activated channels by examining the relative orientation of the voltage-sensor and pore domains. The opening of channels engineered to contain single cysteine residues at the extracellular ends of the voltage-sensing S4 (V246C) and pore-forming S5 (C303) domains is inhibited by formation of disulfide or cysteine:Cd2+ bonds. As Cd2+ coordination is promoted by depolarization, the S4-S5 interaction occurs preferentially in the closed state. The failure of oxidation to catalyze dimer formation, as assayed by Western blotting, indicates the V246C:C303 interaction occurs within a subunit. Intriguingly, a similar interaction has been observed in depolarization-activated Shaker Kv channels at depolarized potentials but such an intrasubunit interaction is inconsistent with the X-ray crystal structure of Kv1.2, wherein S4 approaches S5 of an adjacent subunit. These findings suggest channels of opposite voltage-sensing polarity adopt a conserved S4-S5 orientation in the depolarized state that is distinct from that trapped upon crystallization. PMID:19034494

  16. An image acquisition and registration strategy for the fusion of hyperpolarized helium-3 MRI and x-ray CT images of the lung

    NASA Astrophysics Data System (ADS)

    Ireland, Rob H.; Woodhouse, Neil; Hoggard, Nigel; Swinscoe, James A.; Foran, Bernadette H.; Hatton, Matthew Q.; Wild, Jim M.

    2008-11-01

    The purpose of this ethics committee approved prospective study was to evaluate an image acquisition and registration protocol for hyperpolarized helium-3 magnetic resonance imaging (3He-MRI) and x-ray computed tomography. Nine patients with non-small cell lung cancer (NSCLC) gave written informed consent to undergo a free-breathing CT, an inspiration breath-hold CT and a 3D ventilation 3He-MRI in CT position using an elliptical birdcage radiofrequency (RF) body coil. 3He-MRI to CT image fusion was performed using a rigid registration algorithm which was assessed by two observers using anatomical landmarks and a percentage volume overlap coefficient. Registration of 3He-MRI to breath-hold CT was more accurate than to free-breathing CT; overlap 82.9 ± 4.2% versus 59.8 ± 9.0% (p < 0.001) and mean landmark error 0.75 ± 0.24 cm versus 1.25 ± 0.60 cm (p = 0.002). Image registration is significantly improved by using an imaging protocol that enables both 3He-MRI and CT to be acquired with similar breath holds and body position through the use of a birdcage 3He-MRI body RF coil and an inspiration breath-hold CT. Fusion of 3He-MRI to CT may be useful for the assessment of patients with lung diseases.

  17. Tonic current through GABAA receptors and hyperpolarization-activated cyclic nucleotide-gated channels modulate resonance properties of rat subicular pyramidal neurons.

    PubMed

    Sah, Nirnath; Sikdar, Sujit K

    2014-07-01

    The subiculum, considered to be the output structure of the hippocampus, modulates information flow from the hippocampus to various cortical and sub-cortical areas such as the nucleus accumbens, lateral septal region, thalamus, nucleus gelatinosus, medial nucleus and mammillary nuclei. Tonic inhibitory current plays an important role in neuronal physiology and pathophysiology by modulating the electrophysiological properties of neurons. While the alterations of various electrical properties due to tonic inhibition have been studied in neurons from different regions, its influence on intrinsic subthreshold resonance in pyramidal excitatory neurons expressing hyperpolarization-activated cyclic nucleotide-gated (HCN) channels is not known. Using pharmacological agents, we show the involvement of α5βγ GABAA receptors in the picrotoxin-sensitive tonic current in subicular pyramidal neurons. We further investigated the contribution of tonic conductance in regulating subthreshold electrophysiological properties using current clamp and dynamic clamp experiments. We demonstrate that tonic GABAergic inhibition can actively modulate subthreshold properties, including resonance due to HCN channels, which can potentially alter the response dynamics of subicular pyramidal neurons in an oscillating neuronal network.

  18. Measuring changes in substrate utilization in the myocardium in response to fasting using hyperpolarized [1-13C]butyrate and [1-13C]pyruvate

    PubMed Central

    Bastiaansen, Jessica A. M.; Merritt, Matthew E.; Comment, Arnaud

    2016-01-01

    Cardiac dysfunction is often associated with a shift in substrate preference for ATP production. Hyperpolarized (HP) 13C magnetic resonance spectroscopy (MRS) has the unique ability to detect real-time metabolic changes in vivo due to its high sensitivity and specificity. Here a protocol using HP [1-13C]pyruvate and [1-13C]butyrate is used to measure carbohydrate versus fatty acid metabolism in vivo. Metabolic changes in fed and fasted Sprague Dawley rats (n = 36) were studied at 9.4 T after tail vein injections. Pyruvate and butyrate competed for acetyl-CoA production, as evidenced by significant changes in [13C]bicarbonate (−48%), [1-13C]acetylcarnitine (+113%), and [5-13C]glutamate (−63%), following fasting. Butyrate uptake was unaffected by fasting, as indicated by [1-13C]butyrylcarnitine. Mitochondrial pseudoketogenesis facilitated the labeling of the ketone bodies [1-13C]acetoacetate and [1-13C]β-hydroxybutyryate, without evidence of true ketogenesis. HP [1-13C]acetoacetate was increased in fasting (250%) but decreased during pyruvate co-injection (−82%). Combining HP 13C technology and co-administration of separate imaging agents enables noninvasive and simultaneous monitoring of both fatty acid and carbohydrate oxidation. This protocol illustrates a novel method for assessing metabolic flux through different enzymatic pathways simultaneously and enables mechanistic studies of the changing myocardial energetics often associated with disease. PMID:27150735

  19. A Novel Carbamoyloxy Arylalkanoyl Arylpiperazine Compound (SKL-NP) Inhibits Hyperpolarization-Activated Cyclic Nucleotide-Gated (HCN) Channel Currents in Rat Dorsal Root Ganglion Neurons.

    PubMed

    Chung, Gehoon; Kim, Tae-Hyung; Shin, Hyewon; Chae, Eunhee; Yi, Hanju; Moon, Hongsik; Kim, Hyun Jin; Kim, Joong Soo; Jung, Sung Jun; Oh, Seog Bae

    2012-08-01

    In this study, we determined mode of action of a novel carbamoyloxy arylalkanoyl arylpiperazine compound (SKL-NP) on hyperpolarization-activated cyclic nucleotide-gated (HCN) channel currents (I(h)) that plays important roles in neuropathic pain. In small or medium-sized dorsal root ganglion (DRG) neurons (<40 µm in diameter) exhibiting tonic firing and prominent I(h), SKL-NP inhibited I(h) and spike firings in a concentration dependent manner (IC(50)=7.85 µM). SKL-NP-induced inhibition of I(h) was blocked by pretreatment of pertussis toxin (PTX) and N-ethylmaleimide (NEM) as well as 8-Br-cAMP, a membrane permeable cAMP analogue. These results suggest that SKL-NP modulates I(h) in indirect manner by the activation of a Gi-protein coupled receptor that decreases intracellular cAMP concentration. Taken together, SKL-NP has the inhibitory effect on HCN channel currents (I(h)) in DRG neurons of rats.

  20. Quantitative chemical exchange saturation transfer with hyperpolarized nuclei (qHyper-CEST): Sensing xenon-host exchange dynamics and binding affinities by NMR

    SciTech Connect

    Kunth, M. Witte, C.; Schröder, L.

    2014-11-21

    The reversible binding of xenon to host molecules has found numerous applications in nuclear magnetic resonance studies. Quantitative characterization of the Xe exchange dynamics is important to understand and optimize the physico-chemical behavior of such Xe hosts, but is often challenging to achieve at low host concentrations. We have investigated a sensitive quantification technique based on chemical exchange saturation transfer with hyperpolarized nuclei, qHyper-CEST. Using simulated signals we demonstrated that qHyper-CEST yielded accurate and precise results and was robust in the presence of large amounts of noise (10%). This is of particular importance for samples with completely unknown exchange rates. Using these findings we experimentally determined the following exchange parameters for the Xe host cryptophane-A monoacid in dimethyl sulfoxide in one type of experiment: the ratio of bound and free Xe, the Xe exchange rate, the resonance frequencies of free and bound Xe, the Xe host occupancy, and the Xe binding constant. Taken together, qHyper-CEST facilitates sensitive quantification of the Xe exchange dynamics and binding to hydrophobic cavities and has the potential to analyze many different host systems or binding sites. This makes qHyper-CEST an indispensable tool for the efficient design of highly specific biosensors.

  1. Pacemaker Activity of the Human Sinoatrial Node: An Update on the Effects of Mutations in HCN4 on the Hyperpolarization-Activated Current

    PubMed Central

    Verkerk, Arie O.; Wilders, Ronald

    2015-01-01

    Since 2003, several loss-of-function mutations in the HCN4 gene, which encodes the HCN4 protein, have been associated with sinus node dysfunction. In human sinoatrial node (SAN), HCN4 is the most abundant of the four isoforms of the HCN family. Tetramers of HCN subunits constitute the ion channels that conduct the hyperpolarization-activated “funny” current (If), which plays an important modulating role in SAN pacemaker activity. Voltage-clamp experiments on HCN4 channels expressed in COS-7, CHO and HEK-293 cells, as well as in Xenopus oocytes have revealed changes in the expression and kinetics of mutant channels, but the extent to which especially the kinetic changes would affect If flowing during a human SAN action potential often remains unresolved. In our contribution to the Topical Collection on Human Single Nucleotide Polymorphisms and Disease Diagnostics, we provide an updated review of the mutation-induced changes in the expression and kinetics of HCN4 channels and provide an overview of their effects on If during the time course of a human SAN action potential, as assessed in simulated action potential clamp experiments. Future research may solve apparent inconsistencies between data from clinical studies and data from in vitro and in silico experiments. PMID:25642760

  2. Hyperpolarized 89Y NMR spectroscopic detection of yttrium ion and DOTA macrocyclic ligand complexation: pH dependence and Y-DOTA intermediates

    NASA Astrophysics Data System (ADS)

    Ferguson, Sarah; Kiswandhi, Andhika; Niedbalski, Peter; Parish, Christopher; Kovacs, Zoltan; Lumata, Lloyd

    Dissolution dynamic nuclear polarization (DNP) is a rapidly emerging physics technique used to enhance the signal strength in nuclear magnetic resonance (NMR) and imaging (MRI) experiments for nuclear spins such as yttrium-89 by >10,000-fold. One of the most common and stable MRI contrast agents used in the clinic is Gd-DOTA. In this work, we have investigated the binding of the yttrium and DOTA ligand as a model for complexation of Gd ion and DOTA ligand. The macrocyclic ligand DOTA is special because its complexation with lanthanide ions such as Gd3+ or Y3+ is highly pH dependent. Using this physics technology, we have tracked the complexation kinetics of hyperpolarized Y-triflate and DOTA ligand in real-time and detected the Y-DOTA intermediates. Different kinds of buffers were used (lactate, acetate, citrate, oxalate) and the pseudo-first order complexation kinetic calculations will be discussed. The authors would like to acknowledge the support by US Dept of Defense Award No. W81XWH-14-1-0048 and Robert A. Welch Foundation Grant No. AT-1877.

  3. Iridium Cyclooctene Complex That Forms a Hyperpolarization Transfer Catalyst before Converting to a Binuclear C–H Bond Activation Product Responsible for Hydrogen Isotope Exchange

    PubMed Central

    2016-01-01

    [IrCl(COE)2]2 (1) reacts with pyridine (py) and H2 to form crystallographically characterized IrCl(H)2(COE)(py)2 (2). 2 undergoes py loss to form 16-electron IrCl(H)2(COE)(py) (3), with equivalent hydride ligands. When this reaction is studied with parahydrogen, 1 efficiently achieves hyperpolarization of free py (and nicotinamide, nicotine, 5-aminopyrimidine, and 3,5-lutudine) via signal amplification by reversible exchange (SABRE) and hence reflects a simple and readily available precatayst for this process. 2 reacts further over 48 h at 298 K to form crystallographically characterized (Cl)(H)(py)(μ-Cl)(μ-H)(κ-μ-NC5H4)Ir(H)(py)2 (4). This dimer is active in the hydrogen isotope exchange process that is used in radiopharmaceutical preparations. Furthermore, while [Ir(H)2(COE)(py)3]PF6 (6) forms upon the addition of AgPF6 to 2, its stability precludes its efficient involvement in SABRE. PMID:27934314

  4. Continuing Appropriations Act, 2011

    THOMAS, 111th Congress

    Rep. Lowey, Nita M. [D-NY-18

    2009-06-26

    09/30/2010 Became Public Law No: 111-242. (PDF) (All Actions) Notes: Continuing appropriations through 12/3/2010. Tracker: This bill has the status Became LawHere are the steps for Status of Legislation:

  5. Continual improvement plan

    NASA Technical Reports Server (NTRS)

    1994-01-01

    NASA's approach to continual improvement (CI) is a systems-oriented, agency-wide approach that builds on the past accomplishments of NASA Headquarters and its field installations and helps achieve NASA's vision, mission, and values. The NASA of the future will fully use the principles of continual improvement in every aspect of its operations. This NASA CI plan defines a systematic approach and a model for continual improvement throughout NASA, stressing systems integration and optimization. It demonstrates NASA's constancy of purpose for improvement - a consistent vision of NASA as a worldwide leader in top-quality science, technology, and management practices. The CI plan provides the rationale, structures, methods, and steps, and it defines NASA's short term (1-year) objectives for improvement. The CI plan presents the deployment strategies necessary for cascading the goals and objectives throughout the agency. It also provides guidance on implementing continual improvement with participation from top leadership and all levels of employees.

  6. Continuing Appropriations Act, 2012

    THOMAS, 112th Congress

    Rep. Graves, Sam [R-MO-6

    2011-07-21

    10/05/2011 Became Public Law No: 112-36. (TXT | PDF) (All Actions) Notes: Continuing appropriations through 11/18/2011. Tracker: This bill has the status Became LawHere are the steps for Status of Legislation:

  7. Continuing Liberal Education.

    ERIC Educational Resources Information Center

    Adams, E. M.; Nord, Warren

    1988-01-01

    Liberal education is as essential as continuing professional education, partly because the humanities and liberal arts are "adult matters." Institutions must (1) raise consciousness about its role, and (2) develop concrete ideas for using university resources effectively. (LB)

  8. Continuing Appropriations Act, 2012

    THOMAS, 112th Congress

    Rep. Aderholt, Robert B. [R-AL-4

    2011-05-26

    09/30/2011 Became Public Law No: 112-33. (PDF) (All Actions) Notes: Continuing appropriations through 10/4/2011. Tracker: This bill has the status Became LawHere are the steps for Status of Legislation:

  9. Continuing Appropriations Act, 2012

    THOMAS, 112th Congress

    Rep. Aderholt, Robert B. [R-AL-4

    2011-05-26

    09/30/2011 Became Public Law No: 112-33. (TXT | PDF) (All Actions) Notes: Continuing appropriations through 10/4/2011. Tracker: This bill has the status Became LawHere are the steps for Status of Legislation:

  10. Business Continuity Management Plan

    DTIC Science & Technology

    2014-12-01

    NAVAL POSTGRADUATE SCHOOL MONTEREY, CALIFORNIA MBA PROFESSIONAL REPORT BUSINESS CONTINUITY MANAGEMENT PLAN December 2014...Information Operations and Reports, 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 22202-4302, and to the Office of Management and Budget...

  11. Continuing Appropriations Act, 2012

    THOMAS, 112th Congress

    Rep. Graves, Sam [R-MO-6

    2011-07-21

    10/05/2011 Became Public Law No: 112-36. (PDF) (All Actions) Notes: Continuing appropriations through 11/18/2011. Tracker: This bill has the status Became LawHere are the steps for Status of Legislation:

  12. Continuing Appropriations Act, 2011

    THOMAS, 111th Congress

    Rep. Lowey, Nita M. [D-NY-18

    2009-06-26

    09/30/2010 Became Public Law No: 111-242. (TXT | PDF) (All Actions) Notes: Continuing appropriations through 12/3/2010. Tracker: This bill has the status Became LawHere are the steps for Status of Legislation:

  13. Continuing Appropriations Resolution, 2015

    THOMAS, 113th Congress

    Rep. Rogers, Harold [R-KY-5

    2014-09-09

    09/19/2014 Became Public Law No: 113-164. (PDF) (All Actions) Notes: Continuing appropriations through 12/11/2014. Tracker: This bill has the status Became LawHere are the steps for Status of Legislation:

  14. Continuing Appropriations Resolution, 2015

    THOMAS, 113th Congress

    Rep. Rogers, Harold [R-KY-5

    2014-09-09

    09/19/2014 Became Public Law No: 113-164. (TXT | PDF) (All Actions) Notes: Continuing appropriations through 12/11/2014. Tracker: This bill has the status Became LawHere are the steps for Status of Legislation:

  15. Symmetric continued fractions

    SciTech Connect

    Panprasitwech, Oranit; Laohakosol, Vichian; Chaichana, Tuangrat

    2010-11-11

    Explicit formulae for continued fractions with symmetric patterns in their partial quotients are constructed in the field of formal power series. Similar to the work of Cohn in 1996, which generalized the so-called folding lemma to {kappa}-fold symmetry, the notion of {kappa}-duplicating symmetric continued fractions is investigated using a modification of the 1995 technique due to Clemens, Merrill and Roeder.

  16. Adaptive continuous twisting algorithm

    NASA Astrophysics Data System (ADS)

    Moreno, Jaime A.; Negrete, Daniel Y.; Torres-González, Victor; Fridman, Leonid

    2016-09-01

    In this paper, an adaptive continuous twisting algorithm (ACTA) is presented. For double integrator, ACTA produces a continuous control signal ensuring finite time convergence of the states to zero. Moreover, the control signal generated by ACTA compensates the Lipschitz perturbation in finite time, i.e. its value converges to the opposite value of the perturbation. ACTA also keeps its convergence properties, even in the case that the upper bound of the derivative of the perturbation exists, but it is unknown.