40 results on '"Kuzio, B."'
Search Results
2. Detection of myocardial cell damage in isolated rat hearts in near-infrared spectroscopy
- Author
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Nighswander-Rempel, S., Shaw, R., Kuzio, B., and Kupriyanov, V.
- Subjects
heart - Published
- 2004
3. pH regulation of K+ efflux from myocytes in isolated rat hearts : 87Rb, 7Li, and 31P NMR studies
- Author
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Kupriyanov, V. V., Xiang, B., Kuzio, B., and Deslauriers, R.
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lithium ion efflux ,intracellular pH ,cation/cation exchanger ,rubidium (potassium) ion efflux - Published
- 1999
4. NIRS imaging of intracellular oxygenation in C56BL6 and Kir6.2−/− mouse hearts under stress
- Author
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Jilkina, O., primary, Glogowski, M., additional, Kuzio, B., additional, Zhilkin, P., additional, and Kupriyanov, V.V., additional
- Published
- 2008
- Full Text
- View/download PDF
5. Potassium fluxes and energy metabolism in beating diabetic rat hearts
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Jilkina, O., primary, Kuzio, B., additional, and Kupriyanov, V.V., additional
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- 2007
- Full Text
- View/download PDF
6. Optical imaging of oxygen saturation in beating KATP-deficient mouse hearts
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Jilkina, O., primary, Glogowski, M., additional, Kuzio, B., additional, Zhilkin, P.A., additional, and Kupriyanov, V.V., additional
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- 2007
- Full Text
- View/download PDF
7. 798 Sarcolemma depolarization eliminates mitochondrial uncoupling by an ATP-sensitive potassium channel opener, P-1075, in rat hearts
- Author
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JILKINA, O, primary, KUZIO, B, additional, GROVER, G, additional, and KUPRIYANOV, V, additional
- Published
- 2003
- Full Text
- View/download PDF
8. Imaging of ischemia and infarction in blood-perfused pig hearts using87Rb MRI
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Kupriyanov, V.V., primary, Xiang, B., additional, Sun, J., additional, Jilkina, O., additional, and Kuzio, B., additional
- Published
- 2002
- Full Text
- View/download PDF
9. pH regulation of K+ efflux from myocytes in isolated rate hearts: Rb, Li, and P NMR studies
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Kupriyanov, V.V., Xiang, B., Kuzio, B., and Deslauriers, R.
- Subjects
Nuclear magnetic resonance spectroscopy -- Usage ,Rubidium -- Physiological aspects ,Ion exchange -- Physiological aspects ,Hydrogen-ion concentration -- Physiological aspects ,Heart muscle -- Physiological aspects ,Biological sciences - Abstract
Nuclear magnetic resonance spectroscopy studies were made on rat cardiac muscles to investigate the effects of changes in pH levels on the unidirectional rates of Rb+ and Li+ efflux from myocytes in isolated rat hearts. Findings have indicated that intercellular alkalosis stimulates Rb+ (K+) and Li+ effluxes by activating a nonselective sarcolemmal K+ (Li+)/cation exchanger or a K+ (Li+)-anion symporter.
- Published
- 1999
10. pH regulation of K+efflux from myocytes in isolated rat hearts:87Rb,7Li, and31P NMR studies
- Author
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Kupriyanov, V. V., primary, Xiang, B., additional, Kuzio, B., additional, and Deslauriers, R., additional
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- 1999
- Full Text
- View/download PDF
11. The Effects of Low-Flow Ischemia on K+Fluxes in Isolated Rat Hearts Assessed by87Rb NMR
- Author
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Kupriyanov, V.V., primary, Xiang, B., additional, Kuzio, B., additional, and Deslauriers, R., additional
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- 1999
- Full Text
- View/download PDF
12. Imaging of ischemia and infarction in blood-perfused pig hearts using 87Rb MRI.
- Author
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Kupriyanov, V.V., Xiang, B., Sun, J., Jilkina, O., and Kuzio, B.
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- 2003
- Full Text
- View/download PDF
13. Imaging of ischemia and infarction in blood-perfused pig hearts using <SUP>87</SUP>Rb MRI
- Author
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Kupriyanov, V.V., Xiang, B., Sun, J., Jilkina, O., and Kuzio, B.
- Abstract
87Rb-MRI was used to measure Rb+ uptake in blood-perfused pig hearts during complete occlusion (120 and 70 min) of the left anterior descending artery (LAD) and subsequent reperfusion (120 and 170 min). The Rb+ uptake rate and maximal Rb image intensity during 120-min occlusion were significantly lower in the ischemic anterior left ventricular (LV) wall (0.35 ± 0.14%/min and 26 ± 4.7%) relative to those in the remote posterior LV wall (2.43 ± 0.33%/min and 98 ± 10%). Reperfusion after 120 and 70 min of occlusion resulted in formation of damaged areas, which had 40 ± 4 and 73 ± 10% of the 87Rb image intensity observed in the remote posterior wall. The infarct sizes determined histologically by triphenyltetrazolium chloride staining were 9.6 ± 3.4 and 5.6 ± 4.6% of the total ventricular mass (LV + RV) in the 120- and 70-min occlusion groups, respectively. The sizes determined by MRI were 13.1 ± 2 and 2.8 ± 4.3% of the total number of pixels, respectively. The Rb+ uptake in the anterior wall during 120-min occlusion was somewhat lower than that previously observed in crystalloid-perfused hearts. It is concluded that blood does not interfere with the ability of 87Rb MRI to detect ischemic and infarct areas. Magn Reson Med 49:99107, 2003. © 2003 Wiley-Liss, Inc.
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- 2003
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14. Imaging of ischemia and infarction in blood‐perfused pig hearts using 87Rb MRI
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Kupriyanov, V.V., Xiang, B., Sun, J., Jilkina, O., and Kuzio, B.
- Abstract
87Rb‐MRI was used to measure Rb+uptake in blood‐perfused pig hearts during complete occlusion (120 and 70 min) of the left anterior descending artery (LAD) and subsequent reperfusion (120 and 170 min). The Rb+uptake rate and maximal Rb image intensity during 120‐min occlusion were significantly lower in the ischemic anterior left ventricular (LV) wall (0.35 ± 0.14%/min and 26 ± 4.7%) relative to those in the remote posterior LV wall (2.43 ± 0.33%/min and 98 ± 10%). Reperfusion after 120 and 70 min of occlusion resulted in formation of damaged areas, which had 40 ± 4 and 73 ± 10% of the 87Rb image intensity observed in the remote posterior wall. The infarct sizes determined histologically by triphenyltetrazolium chloride staining were 9.6 ± 3.4 and 5.6 ± 4.6% of the total ventricular mass (LV + RV) in the 120‐ and 70‐min occlusion groups, respectively. The sizes determined by MRI were 13.1 ± 2 and 2.8 ± 4.3% of the total number of pixels, respectively. The Rb+uptake in the anterior wall during 120‐min occlusion was somewhat lower than that previously observed in crystalloid‐perfused hearts. It is concluded that blood does not interfere with the ability of 87Rb MRI to detect ischemic and infarct areas. Magn Reson Med 49:99–107, 2003. © 2003 Wiley‐Liss, Inc.
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- 2003
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15. 798 Sarcolemma depolarization eliminates mitochondrial uncoupling by an ATP-sensitive potassium channel opener, P-1075, in rat hearts
- Author
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Jilkina, O., Kuzio, B., Grover, G., and Kupriyanov, V.
- Subjects
- *
SARCOLEMMA , *POTASSIUM channels - Abstract
An abstract of the study "Sarcolemma depolarization eliminates mitochondrial uncoupling by an ATP-sensitive potassium channel opener, P-1075, in rat hearts," by O. Jilkina et al. is presented.
- Published
- 2004
16. NIRS imaging of intracellular oxygenation in C56BL6 and Kir6.2−/− mouse hearts under stress
- Author
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Jilkina, O., Glogowski, M., Kuzio, B., Zhilkin, P., and Kupriyanov, V.V.
- Published
- 2008
- Full Text
- View/download PDF
17. Optical imaging of oxygen saturation in beating KATP-deficient mouse hearts
- Author
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Jilkina, O., Glogowski, M., Kuzio, B., Zhilkin, P.A., and Kupriyanov, V.V.
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- 2007
- Full Text
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18. DNA-PK hyperactivation occurs in deletion 11q chronic lymphocytic leukemia and is both a biomarker and therapeutic target for drug-resistant disease.
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Kost SEF, Saleh A, Yuan SH, Kuzio B, Gibson SB, Yang L, Banerji V, Johnston JB, and Katyal S
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- Humans, Biomarkers, Chromosome Deletion, DNA, DNA-Activated Protein Kinase metabolism, DNA-Binding Proteins metabolism, Prognosis, Drug Resistance, Neoplasm, Chromosomes, Human, Pair 11, Leukemia, Lymphocytic, Chronic, B-Cell drug therapy, Leukemia, Lymphocytic, Chronic, B-Cell genetics
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- 2023
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19. RAD51-Mediated DNA Homologous Recombination Is Independent of PTEN Mutational Status.
- Author
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Sinha A, Saleh A, Endersby R, Yuan SH, Chokshi CR, Brown KR, Kuzio B, Kauppinen T, Singh SK, Baker SJ, McKinnon PJ, and Katyal S
- Abstract
PTEN mutation occurs in a variety of aggressive cancers and is associated with poor patient outcomes. Recent studies have linked mutational loss of PTEN to reduced RAD51 expression and function, a key factor involved in the homologous recombination (HR) pathway. However, these studies remain controversial, as they fail to establish a definitive causal link to RAD51 expression that is PTEN-dependent, while other studies have not been able to recapitulate the relationship between the PTEN expression and the RAD51/HR function. Resolution of this apparent conundrum is essential due to the clinically-significant implication that PTEN-deficient tumors may be sensitive to poly (ADP-ribose) polymerase (PARP) inhibitors (PARPi) commonly used in the clinical management of BRCA -mutated and other HR-deficient (HRD) tumors., Methods: Primary Pten -deficient (and corresponding wild-type) mouse embryonic fibroblasts (MEFs) and astrocytes and PTEN -null human tumor cell lines and primary cells were assessed for RAD51 expression (via the Western blot analysis) and DNA damage repair analyses (via alkali comet and γH2AX foci assays). RAD51 foci analysis was used to measure HR-dependent DNA repair. Xrcc2 -deficient MEFs served as an HR-deficient control, while the stable knockdown of RAD51 ( shRAD51 ) served to control for the relative RAD51/HR-mediated repair and the phospho-53BP1 foci analysis served to confirm and measure non-homologous end joining (NHEJ) activity in PTEN-deficient and shRAD51 -expressing (HRD) lines. Cell proliferation studies were used to measure any potential added sensitivity of PTEN -null cells to the clinically-relevant PARPi, olaparib. RAD51 levels and DNA damage response signaling were assessed in PTEN-mutant brain tumor initiating cells (BTICs) derived from primary and recurrent glioblastoma multiforme (GBM) patients, while expression of RAD51 and its paralogs were examined as a function of the PTEN status in the RNA expression datasets isolated from primary GBM tumor specimens and BTICs., Results: Pten knockout primary murine cells display unaltered RAD51 expression, endogenous and DNA strand break-induced RAD51 foci and robust DNA repair activity. Defective HR was only observed in the cells lacking Xrcc2 . Likewise, human glioblastoma multiforme (GBM) cell lines with known PTEN deficiency (U87, PTEN -mutated; U251 and U373, PTEN -null) show apparent expression of RAD51 and display efficient DNA repair activity. Only GBM cells stably expressing shRNAs against RAD51 ( shRAD51 ) display dysfunctional DNA repair activity and reduced proliferative capacity, which is exacerbated by PARPi treatment. Furthermore, GBM patient-derived BTICs displayed robust RAD51 expression and intact DNA damage response signaling in spite of PTEN -inactivating mutations. RNA expression analysis of primary GBM tissue specimens and BTICs demonstrate stable levels of RAD51 and its paralogs ( RAD51B, RAD51C, RAD51D, XRCC2, XRCC3 , and DMC1 ), regardless of the PTEN mutational status., Conclusions: Our findings demonstrate definitively that PTEN loss does not alter the RAD51 expression, its paralogs, or the HR activity. Furthermore, deficiency in PTEN alone is not sufficient to impart enhanced sensitivity to PARPi associated with HRD. This study is the first to unequivocally demonstrate that PTEN deficiency is not linked to the RAD51 expression or the HR activity amongst primary neural and non-neural Pten -null cells, PTEN-deficient tumor cell lines, and primary PTEN -mutant GBM patient-derived tissue specimens and BTICs.
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- 2020
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20. Near-infrared fluorescence imaging of mouse myocardial microvascular endothelium using Cy5.5-lectin conjugate.
- Author
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Nguyen C, Bascaramurty S, Kuzio B, Gregorash L, Kupriyanov V, and Jilkina O
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- Animals, Female, Fluorescent Dyes metabolism, Male, Mice, Mice, Inbred C57BL, Perfusion, Carbocyanines metabolism, Coronary Vessels cytology, Endothelium, Vascular cytology, Endothelium, Vascular metabolism, Microvessels cytology, Optical Imaging methods, Plant Lectins metabolism
- Abstract
Cy5.5-lectin, a non-toxic conjugate, combines the benefits of near-infrared (NIR) imaging, such as significant reduction of background fluorescence and increased tissue depth penetration, with its affinity for vascular endothelial cells. When compared to endothelial staining methods using FITC-lectin and ICAM2 antibodies, Cy5.5-lectin was confirmed to specifically bind endothelial cells and produce a fluorescence signal both in real-time and post-infusion. Ex-vivo experiments with isolated hearts demonstrated that binding was limited to perfused areas of the myocardium. With mouse in-vivo tail-vein injections, other organs such as the liver, spleen, and kidney were also stained and yielded similar quality images of the heart., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2012
- Full Text
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21. Defects in myoglobin oxygenation in K(ATP)-deficient mouse hearts under normal and stress conditions characterized by near infrared spectroscopy and imaging.
- Author
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Jilkina O, Glogowski M, Kuzio B, Zhilkin PA, Gussakovsky E, and Kupriyanov VV
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- 2,4-Dinitrophenol pharmacology, Acute Disease, Animals, Cardiomyopathy, Dilated genetics, Cardiotonic Agents pharmacology, Female, In Vitro Techniques, Isoproterenol pharmacology, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Models, Biological, Potassium Channels, Inwardly Rectifying genetics, Spectroscopy, Near-Infrared methods, Stress, Physiological drug effects, Uncoupling Agents pharmacology, Vasodilation drug effects, Cardiomyopathy, Dilated metabolism, Myocardium metabolism, Myoglobin metabolism, Oxygen metabolism, Potassium Channels, Inwardly Rectifying metabolism, Stress, Physiological physiology
- Abstract
Background: Disruption of ATP-sensitive potassium (K(ATP)) channel activity results in the development of dilated cardiomyopathy in response to different forms of stress, likely due to the underlying metabolic defects. To further understand the role of Kir6.2-containing channels in the development of cardiac disease, we analysed the left ventricular (LV) wall oxygenation and the physiologic responses induced by acute stress in non-dilated Kir6.2(-/-) hearts., Methods: Control (C57BL6) and Kir6.2(-/-) mouse hearts were perfused in constant flow Langendorff mode with Krebs-Henseleit buffer. Myocardial oxygenation was evaluated using a newly developed technique, near infrared spectroscopic imaging (NIRSI) of the myoglobin (Mb) oxygen saturation parameter (OSP, ratio of oxy- to total Mb)., Results: 2,4-dinitrophenol (DNP, 50-µM) and isoproterenol (0.1-µM) failed to produce a transient vasodilatory response and caused a significant diastolic pressure increase in Kir6.2(-/-) hearts. DNP strongly suppressed contractile function in both groups and induced severe mean OSP decreases in Kir6.2(-/-) hearts. Isoproterenol-induced decreases in OSP were similar despite the lack of contractile function stimulation in the Kir6.2(-/-) group. The index of OSP spatial heterogeneity (relative dispersion, RD) was lower by 15% in the Kir6.2(-/-) group at the baseline conditions. Recovery after stress caused reduction of RD values by 20% (DNP) and 8% (isoproterenol) in controls; however, these values did not change in the Kir6.2(-/-) group., Conclusions: 1) NIRSI can be used to analyse 2-D dynamics of LV oxygenation in rodent models of cardiomyopathy; 2) Kir6.2-containing K(ATP) channels play an important role in maintaining myocardial oxygenation balance under acute stress conditions and in post-stress recovery., (Crown Copyright © 2010. Published by Elsevier Ireland Ltd. All rights reserved.)
- Published
- 2011
- Full Text
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22. Rhodamine 800 as a near-infrared fluorescent deposition flow tracer in rodent hearts.
- Author
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Munch G, McKay S, Gussakovsky E, Kuzio B, Kupriyanov VV, and Jilkina O
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- Animals, Contrast Media pharmacokinetics, Female, Image Processing, Computer-Assisted, Kinetics, Least-Squares Analysis, Male, Mitochondria, Heart chemistry, Mitochondria, Heart metabolism, Myocardial Ischemia metabolism, Myocardial Ischemia pathology, Myocardium metabolism, Rats, Rats, Inbred WKY, Rhodamines pharmacokinetics, Contrast Media chemistry, Myocardium chemistry, Rhodamines chemistry, Spectrometry, Fluorescence methods, Spectroscopy, Near-Infrared methods
- Abstract
We investigated the use of a near-infrared (NIR) fluorescent dye, Rhodamine 800 (Rhod800, λ(exc) = 693 nm, λ(em) > 720 nm) as a flow-dependent molecular tracer for NIR spectroscopy and high-resolution cardiac imaging. Rhod800 accumulates in isolated mitochondria in proportion to the mitochondrial membrane potential (ΔΨ). However, in the intact myocardium, Rhod800 binding is ΔΨ-independent. Rat hearts were perfused in a Langendorff mode with Krebs-Henseleit buffer containing 45-nM Rhod800 at normal (100%), increased (150%), or reduced (50%) baseline coronary flow (CF) per gram, for 30 to 60 min. In a different group of hearts, the left anterior descending artery (LAD) was occluded prior to Rhod800 infusion to create a flow deficit area. Rhod800 deposition was analyzed by: 1. absorbance spectroscopy kinetics in the Rhod800-perfused hearts, 2. Rhod800 absorbance and fluorescence imaging in the short-axis heart slices, and 3. dynamic epicardial/subepicardial fluorescence imaging of Rhod800 in KCl-arrested hearts, with a spatial resolution of ∼ 200 μm. Rhod800 deposition was proportional to the perfusate volume (CF and perfusion time) and there was no Rhod800 loss during the washout period. In the LAD-ligated hearts, Rhod800 fluorescence was missing from the no-flow, LAD-dependent endocardial and epicardial/subepicardial area. We concluded that Rhod800 can be used as a deposition flow tracer for dynamic cardiac imaging.
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- 2011
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23. Fluorescence imaging to quantify the fluorescent microspheres in cardiac tissue.
- Author
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Gussakovsky E, Kuzio B, Yang Y, and Kupriyanov V
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- Animals, Fluorescence, Myocardium metabolism, Regional Blood Flow, Sensitivity and Specificity, Swine, Imaging, Three-Dimensional, Microspheres, Myocardium pathology, Spectrometry, Fluorescence methods
- Abstract
To quantify the fluorescent microsphere (FM) content in cardiac tissue, which is an indicative of blood flow, fluorescence imaging of both sides of the pig heart slice was employed. Despite the light scattering inside the tissue and contributions from multiple tissue layers to the total emission, it is shown that the fluorescence intensity at any pixel is proportional to the FM content and the fluorescence image may be transformed to the image of the FM concentration. A convenient standard for the emission-FM concentration transformation is proposed. The approach has several advantages in comparison with the traditional "digestion & extraction" method such as: non-destructiveness, high spatial resolution, high throughput, repeatability and simplicity of operation., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
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- 2011
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24. Potassium fluxes, energy metabolism, and oxygenation in intact diabetic rat hearts under normal and stress conditions.
- Author
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Jilkina O, Kuzio B, and Kupriyanov VV
- Subjects
- 2,4-Dinitrophenol pharmacology, Adrenergic beta-Agonists pharmacology, Algorithms, Animals, Diabetes Mellitus, Experimental metabolism, Guanidines pharmacology, Isoproterenol pharmacology, KATP Channels metabolism, Magnetic Resonance Spectroscopy, Male, Phosphorus Isotopes chemistry, Pyridines pharmacology, Rats, Rats, Sprague-Dawley, Rubidium Radioisotopes, Sarcolemma drug effects, Sarcolemma metabolism, Sodium-Potassium-Exchanging ATPase metabolism, Diabetes Mellitus, Experimental physiopathology, Energy Metabolism physiology, Heart physiopathology, Myocardium metabolism, Oxygen Consumption physiology, Potassium metabolism, Stress, Physiological drug effects
- Abstract
We evaluated the function of Na(+)/K(+) ATPase and sarcolemmal K(ATP) channels in diabetic rat hearts. Six weeks after streptozotocin (STZ) injection, unidirectional K(+) fluxes were assayed by using (87)rubidium ((87)Rb(+)) MRS. The hearts were loaded with Rb(+) by perfusion with Krebs-Henseleit buffer, in which 50% of K(+) was substituted with Rb(+). The rate constant of Rb(+) uptake via Na(+)/K(+) ATPase was reduced. K(ATP)-mediated Rb(+) efflux was activated metabolically with 2,4-dinitrophenol (DNP, 50 micromol.L(-1)) or pharmacologically with a K(ATP) channel opener, P-1075 (5 micromol.L(-1)). Cardiac energetics were monitored by using (31)P MRS and optical spectroscopy. DNP produced a smaller ATP decrease, yet similar Rb(+) efflux activation in STZ hearts. In K(+)-arrested hearts, P-1075 had no effect on high-energy phosphates and stimulated Rb(+) efflux by interaction with SUR2A subunit of K(ATP) channel; this stimulation was greater in STZ hearts. In normokalemic hearts, P-1075 caused cardiac arrest and ATP decline, and the stimulation of Rb(+) efflux was lower in normokalemic STZ hearts arrested by P-1075. Thus, the Rb(+)efflux stimulation in STZ hearts was altered depending on the mode of K(ATP) channel activation: pharmacologic stimulation (P-1075) was enhanced, whereas metabolic stimulation (DNP) was reduced. Both the basal concentration of phosphocreatine ([PCr]) and [PCr]/[ATP] were reduced; nevertheless, the STZ hearts were more or equally resistant to metabolic stress.
- Published
- 2008
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25. K+ transport and energetics in Kir6.2(-/-) mouse hearts assessed by 87Rb and 31P magnetic resonance and optical spectroscopy.
- Author
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Jilkina O, Kuzio B, Rendell J, Xiang B, and Kupriyanov VV
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- 2,4-Dinitrophenol pharmacology, Animals, Electron Transport Complex IV metabolism, In Vitro Techniques, Mice, Mice, Inbred C57BL, Mice, Knockout, Myoglobin metabolism, Perfusion methods, Phosphates metabolism, Potassium Channels, Inwardly Rectifying genetics, Spectrum Analysis methods, Magnetic Resonance Spectroscopy methods, Myocardial Contraction, Myocardium metabolism, Potassium Channels physiology, Potassium Channels, Inwardly Rectifying physiology
- Abstract
Cardiac sarcolemmal K(ATP) channels are crucial in adaptation to stress caused by metabolic inhibition and moderate exercise, which requires not only down-regulation of energy spending, but also up-regulation of mitochondrial ATP synthesis. To investigate sarcolemmal and mitochondrial effects of a Kir6.2 (K(+) ion-selective subunit of the channel) knockout, we used non-invasive techniques ((87)Rb, (31)P NMR and optical spectroscopy) to study (1) K(+) fluxes, (2) high-energy phosphates, (3) the cytochrome c oxidase redox state, (4) myoglobin deoxygenation, and (5) contractile function at the baseline and in response to metabolic uncoupling with 2,4-dintrophenol (DNP) and stimulation with isoproterenol in Langendorff-perfused mouse hearts. Comparison with control C57BL6 hearts demonstrated that the Kir6.2 knockout resulted in: (a) a lack of stimulation of the unidirectional potassium efflux from the hearts when K(ATP) channels were activated metabolically by DNP (50 muM, 20 min); (b) a decrease in ATP, but not phosphocreatine, at the baseline, that became even more pronounced when the hearts were subjected to stress due to metabolic inhibition or increased workload caused by isoproterenol infusion (0.1 microM, 20 min); (c) significantly higher reduction of cytochrome c oxidase in response to DNP uncoupling; (d) a blunted response to isoproterenol stimulation. Thus Kir6.2 knockout is associated with decreased tolerance of mouse hearts to metabolic inhibition and catecholamine stress.
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- 2006
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26. Interaction of a mitochondrial membrane potential-sensitive dye, rhodamine 800, with rat mitochondria, cells, and perfused hearts.
- Author
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Jilkina O, Kong HJ, Hwi L, Kuzio B, Xiang B, Manley D, Jackson M, and Kupriyanov VV
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- Animals, Cells, Cultured, Fluorescent Dyes pharmacokinetics, Male, Metabolic Clearance Rate, Organ Specificity, Perfusion, Rats, Rats, Sprague-Dawley, Tissue Distribution, Hepatocytes metabolism, Membrane Potentials physiology, Microscopy, Fluorescence methods, Mitochondria, Liver metabolism, Myocytes, Cardiac metabolism, Rhodamines pharmacokinetics, Spectrometry, Fluorescence methods
- Abstract
Fluorescence, absorbance, and binding of a mitochondrial membrane potential-sensitive probe, rhodamine 800 (rhod800), were measured in isolated rat mitochondria, hepatocytes, cardiomyocytes, and hearts in the presence or absence of mitochondrial uncouplers. Excitation of rhod800 was achieved with laser diodes (690 or 670 nm) and resulted in a fluorescence peak at 720 nm. Greater than 99% of rhod800 (1 microM) was taken up from the buffer by energized mitochondria. This resulted in a fluorescence decrease by 77% (13% in de-energized mitochondria). Sixty-seven percent of rhod800 was taken up by cardiomyocytes and 75% by hepatocytes resulting in the fluorescence decrease by 16% and 37%, respectively, which were reversed by approximately 10% upon cell uncoupling. In hearts, binding, absorbance, and fluorescence were almost uncoupler-insensitive possibly due to rhod800 interaction outside of mitochondria. Fluorescence of the hearts perfused with 27.5 and 55 nM rhod800 was measured in orthogonal and reflection modes. The former provided deep tissue penetration (approximately a centimeter); however, nonlinearity between absorbance and fluorescence was evident. In the latter setting, depth of tissue penetration was approximately a millimeter, which eliminated an inner filter effect and restored linearity. We concluded that excessive hydrophobicity of rhod800 complicates detection of energy-dependent fluorescence changes in myocardium.
- Published
- 2006
- Full Text
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27. Potassium transport in Langendorff-perfused mouse hearts assessed by 87Rb NMR spectroscopy.
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Jilkina O, Xiang B, Kuzio B, Rendell J, and Kupriyanov VV
- Subjects
- Analysis of Variance, Animals, Coronary Circulation, Ion Transport, Mice, Phosphocreatine pharmacology, Rats, Rubidium Radioisotopes, Sodium-Potassium-Exchanging ATPase metabolism, Magnetic Resonance Spectroscopy methods, Myocardium metabolism, Potassium metabolism
- Abstract
We studied the fluxes of a potassium congener (Rb(+)) in mouse hearts by (87)Rb MRS at 8.4T. The hearts were loaded with Rb(+) by perfusion with Krebs-Henseleit buffer, in which 50% of K(+) was substituted with Rb(+). We initiated Rb(+) efflux by changing the perfusion medium to Rb(+)-free buffer. Spectra were acquired every 1.85 min, and the kinetics of Rb(+) transport were analyzed by means of monoexponential fits. The rate constants of Rb(+) uptake and efflux were 0.0680 +/- 0.0028 and 0.0510 +/- 0.0051 min(-1), respectively (approximately 30% faster than in the rat heart). The ATP-sensitive potassium channel opener, P-1075 (5 microM), and mitochondrial uncoupler, 2,4-dintrophenol (50 microM), activated Rb(+) efflux from mouse hearts by approximately 35%. The mechanisms responsible for the differences in Rb(+) uptake and efflux under baseline conditions and stimulation, in comparison with rat hearts, are discussed. These data provide a background for studies of cardiac potassium transport in transgenic mouse strains., (Copyright 2005 Wiley-Liss, Inc.)
- Published
- 2005
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28. Detection of myocardial cell damage in isolated rat hearts with near-infrared spectroscopy.
- Author
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Nighswander-Rempel SP, Shaw RA, Kuzio B, and Kupriyanov VV
- Subjects
- Animals, Biomarkers analysis, Biomarkers metabolism, Digitonin, Heart drug effects, In Vitro Techniques, Membrane Fluidity drug effects, Metabolic Clearance Rate, Myocardial Stunning chemically induced, Myocardium metabolism, Myoglobin analysis, Rats, Rats, Sprague-Dawley, Image Interpretation, Computer-Assisted methods, Myocardial Stunning diagnosis, Myocardial Stunning metabolism, Myoglobin metabolism, Neodymium, Spectrophotometry, Infrared methods
- Abstract
One hallmark of cell death resulting from prolonged ischemia is cell membrane disruption. We apply optical spectroscopy to gauge membrane disruption in isolated rat hearts by monitoring (1) the washout of myoglobin (Mb) and (2) the accumulation of an exogenous contrast agent in permeabilized cells. The contrast agent, a neodymium (Nd) chelate, has several absorptions in the visible and near-IR, and when present in the perfusate, it cannot penetrate cellular membranes. When membrane integrity is disrupted, however, it is expected to accumulate within the intracellular space; moreover, cellular Mb is expected to wash out. To test this hypothesis, rat hearts (n=12) are perfused with Krebs-Henseleit buffer (KHB), followed by perfusion with KHB in which a 5 mM Nd-DTPA solution is present. Membrane damage is then induced by infusion of digitonin into the Nd-KHB perfusate to provide a digitonin concentration of 2.5, 5, or 10 microg/mL. After 30 min of infusion, Mb levels fall to 46+/-14% of baseline levels and Nd-DTPA rises to 161+/-19% of predigitonin levels. No apparent dependence of total membrane disruption on digitonin concentration over the concentration range studied is found, although higher concentrations do lead to more rapid membrane disruption.
- Published
- 2004
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29. Sarcolemmal and mitochondrial effects of a KATP opener, P-1075, in "polarized" and "depolarized" Langendorff-perfused rat hearts.
- Author
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Jilkina O, Kuzio B, Grover GJ, Folmes CD, Kong HJ, and Kupriyanov VV
- Subjects
- Animals, Heart Arrest, Induced, Liver metabolism, Male, Membrane Potentials, Potassium metabolism, Rats, Rats, Sprague-Dawley, Rubidium metabolism, Guanidines pharmacology, Heart drug effects, Mitochondria drug effects, Pyridines pharmacology, Sarcolemma drug effects, Vasodilator Agents pharmacology
- Abstract
We investigated consequences of cardiac arrest on sarcolemmal and mitochondrial effects of ATP-sensitive potassium channel (KATP) opener, P-1075, in Langendorff-perfused rat hearts. Depolarised cardiac arrest (24.7 mM KCl) did not affect glibenclamide-sensitive twofold activation of rubidium efflux by P-1075 (5 microM) from rubidium-loaded hearts, but eliminated uncoupling produced by P-1075 in beating hearts: 40% depletion of phosphocreatine and ATP, 50% increase in oxygen consumption, and reduction of cytochrome c oxidase. Depolarized cardiac arrest by calcium channel blocker, verapamil (5 microM), also prevented uncoupling. Lack of P-1075 mitochondrial effects in depolarized hearts was not due to changes in phosphorylation potential, because 2,4-dintrophenol (10 microM) reversed the [PCr]/[Cr] increase and Pi decrease, characteristic of KCl-arrest, but did not restore uncoupling. In agreement with this conclusion, pyruvate (5 mM) increased [PCr]/[Cr] and decreased Pi, but did not prevent uncoupling in beating hearts. A decrease in mean [Ca2+] in KCl-arrested hearts could not account for lack of P-1075 mitochondrial effects, because calcium channel opener, S-(-)-Bay K8644 (50 nM), and beta-agonist, isoproterenol (0.5 microM), did not facilitate uncoupling. In contrast, in adenosine (1 mM)-arrested hearts (polarized arrest), P-1075 caused 40% phosphocreatine and ATP depletion. In isolated rat liver mitochondria, P-1075 (20 microM) decreased mitochondrial membrane potential (DeltaPsi) by approximately 14 mV (demonstrated by redistribution of DeltaPsi-sensitive dye, rhodamine 800) in a glibenclamide-sensitive manner. We concluded that cell membrane depolarization does not prevent activation of sarcolemmal KATP by P-1075, but it plays a role in mitochondrial uncoupling effects of P-1075.
- Published
- 2003
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30. Cardioselective sulfonylthiourea HMR 1098 blocks mitochondrial uncoupling induced by a KATP channel opener, P-1075, in beating rat hearts.
- Author
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Jilkina O, Kuzio B, Grover GJ, and Kupriyanov VV
- Subjects
- 2,4-Dinitrophenol pharmacology, Adenosine Triphosphate metabolism, Animals, Electron Transport Complex IV drug effects, Electron Transport Complex IV metabolism, Kinetics, Male, Membrane Proteins metabolism, Mitochondria, Heart drug effects, Oxidative Phosphorylation drug effects, Oxygen Consumption drug effects, Phosphates metabolism, Phosphocreatine metabolism, Potassium Channels, Rats, Rats, Sprague-Dawley, Sarcolemma drug effects, Sarcolemma metabolism, Sulfonylurea Compounds pharmacology, Thiourea pharmacology, Uncoupling Agents pharmacology, Benzamides pharmacology, Guanidines pharmacology, Membrane Proteins pharmacology, Mitochondria, Heart metabolism, Potassium Channel Blockers pharmacology, Pyridines pharmacology, Thiourea analogs & derivatives
- Abstract
We investigated effects of blockade of cardiac ATP-sensitive potassium channels (KATP) with a novel cardioselective sulfonylthiourea, HMR 1098, on metabolic uncoupling caused by a potent KATP opener, P-1075, in Langendorff-perfused rat hearts. We used (1) 87Rb-NMR to detect activation-deactivation of sarcolemmal KATP, (2) 31P-NMR to monitor high-energy phosphates, (3) oxygen uptake measurements to monitor cellular respiration, and (4) myocardial optical absorbance measurements at 603 nm to follow changes in cytochrome c oxidase redox state. Activation of sarcolemmal KATP by P-1075 (5 microM) and a mitochondrial uncoupler 2,4-dinitrophenol (DNP) (50 microM) stimulated Rb+ efflux from the hearts by 130% and 60%, respectively. HMR 1098 (5 and 30 microM) blocked activation of sarcolemmal KATP in situ. HMR 1098 also prevented cardiac arrest and mitochondrial uncoupling induced by P-1075, such as (a) depletion of phosphocreatine and ATP by 40%, (b) two-fold decrease in venous oxygen, and (c) reduction of cytochrome c oxidase (demonstrated by an increase in 603 nm optical absorbance). The metabolic effects of P-1075 can be readily explained by activation of putative mitochondrial KATP. We concluded that blockade of mitochondrial uncoupling by HMR 1098 included an inhibiting effect of HMR 1098 on sarcolemmal and mitochondrial KATP in beating rat hearts.
- Published
- 2003
- Full Text
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31. Hyposmotic shock: effects on rubidium/potassium efflux in normal and ischemic rat hearts, assessed by 87Rb and 31P NMR.
- Author
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Jilkina O, Kuzio B, and Kupriyanov VV
- Subjects
- Animals, Heart physiopathology, Hydrogen-Ion Concentration, In Vitro Techniques, Kinetics, Magnetic Resonance Spectroscopy, Male, Osmotic Pressure, Perfusion methods, Phosphorus Isotopes, Potassium chemistry, Potassium Channel Blockers pharmacology, Rats, Rats, Sprague-Dawley, Rubidium chemistry, Rubidium Radioisotopes, Sodium Potassium Chloride Symporter Inhibitors, Myocardial Ischemia metabolism, Potassium metabolism, Rubidium metabolism
- Abstract
The study evaluated effects of hyposmotic shock on the rate of Rb(+)/K(+) efflux, intracellular pH and energetics in Langendorff-perfused rat hearts with the help of 87Rb- and 31P-NMR. Two models of hyposmotic shock were compared: (1) normosmotic hearts perfused with low [NaCl] (70 mM) buffer, (2) hyperosmotic hearts equilibrated with additional methyl alpha-D-glucopyranoside (Me-GPD, 90 or 33 mM) or urea (90 mM) perfused with normosmotic buffer. Four minutes after hyposmotic shock, Rb(+) efflux rate constant transiently increased approximately two-fold, while pH transiently decreased by 0.08 and 0.06 units, in the first and the second models, respectively, without significant changes in phosphocreatine and ATP. Hyposmotic shock (second model) did not change the rate of Rb(+)/K(+) uptake, indicating that the activity of Na(+)/K(+) ATPase was not affected. Dimethylamiloride (DMA) (10 microM) abolished activation of the Rb(+)/K(+) efflux in the second model; however, Na(+)/H(+) exchanger was not involved, because intracellular acidosis induced by the hyposmotic shock was not enhanced by DMA treatment. After 12 or 20 min of global ischemia, the rate of Rb(+)/K(+) efflux increased by 120%. Inhibitor of the ATP-sensitive potassium channels, glibenclamide (5 microM), partially (40%) decreased the rate constant; however, reperfusion with hyperosmolar buffer (90 mM Me-GPD) did not. We concluded that the shock-induced stimulation of Rb(+)/K(+) efflux occurred, at least partially, through the DMA-sensitive cation/H(+) exchanger and swelling-induced mechanisms did not considerably contribute to the ischemia-reperfusion-induced activation of Rb(+)/K(+) efflux.
- Published
- 2003
- Full Text
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32. Effects of K(ATP) channel openers, P-1075, pinacidil, and diazoxide, on energetics and contractile function in isolated rat hearts.
- Author
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Jilkina O, Kuzio B, Grover GJ, and Kupriyanov VV
- Subjects
- 2,4-Dinitrophenol metabolism, Adenosine Triphosphate metabolism, Animals, In Vitro Techniques, Magnetic Resonance Spectroscopy, Male, Myocardium metabolism, Oxidative Phosphorylation, Oxygen Consumption, Perfusion, Phosphocreatine metabolism, Potassium Channel Blockers metabolism, Potassium Channels drug effects, Rats, Rats, Sprague-Dawley, Vasodilator Agents pharmacology, Diazoxide pharmacology, Guanidines pharmacology, Heart drug effects, Myocardial Contraction drug effects, Pinacidil pharmacology, Potassium Channels metabolism, Pyridines pharmacology
- Abstract
We investigated the metabolic effects of a potent opener of ATP-sensitive K(+) (K(ATP)) channels, P-1075, in perfused rat hearts with the help of(31)P NMR spectroscopy. A 20 min infusion of 5 microm P-1075 depleted phosphocreatine and ATP by approximately 40%, concomitantly with a two-fold increase in inorganic phosphate, while oxygen consumption by the hearts increased by 50%. P-1075 induced a cardiac contracture (left ventricular end diastolic pressure increased from 6 to 60 mmHg) and a cardiac arrest after an infusion of approximately 9 min. The effects were fully reversed by glibenclamide (5 microm), but not by sodium 5-hydroxydecanoate (0.4 m m). A P-1075-related K(ATP) opener, pinacidil (0.3 m m), partially reversed the effects of P-1075, but a structurally unrelated opener, diazoxide (0.5 m m), had no effect. Pinacidil and diazoxide alone did not significantly affect PCr and ATP levels. Bioenergetic and functional effects similar to those of P-1075 were induced by infusion of a classic mitochondrial uncoupler, 2,4-dinitrophenol (50 microm); however, they were not abolished by glibenclamide. In addition, it was shown, using(87)Rb NMR, that both agents, P-1075 and 2,4-dinitrophenol, resulted in a stimulation of Rb(+) efflux from the Rb(+) loaded rat hearts by approximately 130 and 65%, respectively, in a glibenclamide-sensitive manner. An inhibitory effect of P-1075 on ATP synthesis cannot be explained by its well-known action on sarcolemmal K(ATP) channels. We concluded that, unlike an uncoupling effect of 2,4-dinitrophenol, an inhibitory effect of P-1075 is produced by uncoupling of oxidative phosphorylation through the activation of mitochondrial K(ATP) channels., (Copyright 2002 Academic Press.)
- Published
- 2002
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33. An inhibitor of poly (ADP-ribose) synthetase activity reduces contractile dysfunction and preserves high energy phosphate levels during reperfusion of the ischaemic rat heart.
- Author
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Docherty JC, Kuzio B, Silvester JA, Bowes J, and Thiemermann C
- Subjects
- Animals, Blood Pressure drug effects, Cardiotonic Agents pharmacology, Diastole, Heart physiopathology, In Vitro Techniques, Isoproterenol pharmacology, Isoquinolines pharmacology, Magnetic Resonance Spectroscopy, Male, Myocardial Ischemia metabolism, Myocardial Reperfusion, Phosphocreatine drug effects, Phosphocreatine metabolism, Rats, Rats, Sprague-Dawley, Systole, Ventricular Dysfunction, Left physiopathology, Adenosine Triphosphate metabolism, Enzyme Inhibitors pharmacology, Heart drug effects, Myocardial Contraction drug effects, Myocardial Ischemia physiopathology, Poly(ADP-ribose) Polymerase Inhibitors
- Abstract
The cardioprotective properties of inhibition of poly (ADP-ribose) synthetase (PARS) were investigated in the isolated perfused heart of the rat. Hearts were perfused in the Langendorff mode and subjected to 23 min total global ischaemia and reperfused for 60 min. Left ventricular function was assessed by means of an intra-ventricular balloon. High energy phosphates were measured by 31P-NMR spectroscopy. Intracellular levels of NAD were measured by capillary electrophoresis of perchloric acid extracts of hearts at the end of reperfusion. Reperfusion in the presence of the PARS inhibitor 1,5 didroxyisoquinoline (ISO, 100 microM) attenuated the mechanical dysfunction observed following 1 h of reperfusion; 27+/-13 and 65+/-8% recovery of preischaemic rate pressure product for control and 100 microM ISO, respectively. This cardioprotection was accompanied by a preservation of intracellular high-energy phosphates during reperfusion; 38+/-2 vs 58+/-4% (P<0.05) of preischaemic levels of phosphocreatine (PCr) for control and 100 microM ISO respectively and 23+/-1 vs 31+/-3% (P < 0.05) of preischaemic levels of ATP for control and 100 microM ISO respectively. Cellular levels of NAD were higher in ISO treated hearts at the end of reperfusion; 2.56+/-0.45 vs 4.76+/-1.12 micromoles g(-1) dry weight (P<0.05) for control and ISO treated. These results demonstrate that the cardioprotection afforded by inhibition of PARS activity with ISO is accompanied by a preservation of high-energy phosphates and cellular NAD levels and suggest that the mechanism responsible for this cardioprotection may involve prevention of intracellular ATP depletion.
- Published
- 1999
- Full Text
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34. pH regulation of K(+) efflux from myocytes in isolated rat hearts: (87)Rb, (7)Li, and (31)P NMR studies.
- Author
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Kupriyanov VV, Xiang B, Kuzio B, and Deslauriers R
- Subjects
- Acidosis metabolism, Alkalosis metabolism, Ammonium Chloride pharmacology, Animals, Hydrogen-Ion Concentration, In Vitro Techniques, Lithium, Magnetic Resonance Spectroscopy, Male, Monensin pharmacology, Phosphorus, Rats, Rats, Sprague-Dawley, Rubidium Radioisotopes, Myocardium cytology, Myocardium metabolism, Potassium metabolism
- Abstract
This study investigates the effects of intracellular (pH(i)) and extracellular pH (pH(e)) on the efflux of Rb(+) and Li(+) in isolated rat hearts. (87)Rb and (7)Li NMR were used to measure Rb(+) and Li(+) content, respectively, of hearts, and (31)P NMR was used to monitor pH(i), pH(e), and phosphate levels. After 30-min equilibration with Rb(+) or Li(+), effluxes were initiated by switching perfusion to a Rb(+)- or Li(+)-free, high-K(+) (20.7 mM) Krebs-Henseleit buffer with 15 microM bumetanide. Monensin (2 microM) increased pH(i) from 7.10 +/- 0.05 to 7.32 +/- 0.07 and resulted in activation of Rb(+) efflux; the first-order rate constant (k x 10(3), in min(-1)) increased from 42 +/- 2 to 116 +/- 16. Glibenclamide (4 microM) did not inhibit monensin-activated Rb(+) efflux (k = 110 +/- 17), whereas quinine (0.2 mM) slightly inhibited it by 19 +/- 9%. Infusion of 15 mM NH(4)Cl during Rb(+) washout increased k for Rb(+) efflux by 93% (81 +/- 8), which was glibenclamide and quinine insensitive, and caused a transient increase in pH(i) to 7.25 +/- 0.08. Intracellular Li(+) inhibited NH(4)Cl-stimulated Rb(+) efflux by 55%. Monensin and NH(4)Cl stimulated Li(+) efflux by 40%, increasing k from 29 +/- 3 to 43 +/- 7 and 41 +/- 3, respectively. The stimulation was not sensitive to 10 microM dimethylamiloride. Intracellular acidosis that resulted from the washout of NH(4)Cl (pH 6.86 +/- 0.2) slightly inhibited Rb(+) efflux (k = 36 +/- 5), whereas NH(4)Cl itself in the absence of pH(i) changes did not markedly affect Rb(+) efflux. A moderate increase in pH(i) (7.17 +/- 0.06) produced by washout of 15 mM 2, 2-dimethylpropionate (DMP)-Tris from hearts preequilibrated with DMP did not markedly affect Rb(+) efflux. Neither global alkalosis (pH(i) 7.4, pH(e) 7.55) nor acidosis (pH(i) approximately pH(e) 6.8) produced by 3 mM Tris base or 5 mM MES, respectively, affected Rb(+) efflux. We suggest that intracellular alkalosis stimulates Rb(+) (K(+)) and Li(+) effluxes by activating a nonselective sarcolemmal K(+) (Li(+))/cation exchanger or a K(+) (Li(+))-anion symporter.
- Published
- 1999
- Full Text
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35. The effects of low-flow ischemia on K+ fluxes in isolated rat hearts assessed by 87Rb NMR.
- Author
-
Kupriyanov VV, Xiang B, Kuzio B, and Deslauriers R
- Subjects
- 2,4-Dinitrophenol pharmacology, Adenosine Triphosphate metabolism, Animals, Coronary Circulation, Energy Metabolism, Hemodynamics, In Vitro Techniques, Ion Transport drug effects, Magnetic Resonance Spectroscopy, Male, Myocardial Ischemia physiopathology, Perfusion, Phosphocreatine metabolism, Potassium Channels metabolism, Rats, Rats, Sprague-Dawley, Rubidium Radioisotopes, Sodium-Potassium-Exchanging ATPase metabolism, Myocardial Ischemia metabolism, Potassium metabolism
- Abstract
This study investigated whether Na+/K+ ATPase is inhibited and KATP channels activated during low flow ischemia (LFI) by monitoring Rb+ uptake and efflux from rat hearts using 87Rb NMR. In the uptake experiments, isolated Langendorff perfused hearts were exposed to Rb+-containing Krebs-Henseleit buffer (2.14 m m+3.76 m m K+) for 60 min. When Rb+ uptake started the flow of perfusate was decreased from 10 to 1 ml/min/g wet weight for 44 min and then returned to normal. The rate of Rb+ uptake and its equilibrium level decreased to 40 and 65% of the control (no ischemia) levels, respectively. Phosphocreatine and cytoplasmic [ATP]/[ADP] measured by 31P NMR decreased by half, intracellular pH (pHi) decreased to 6.8+/-0.1, and Pi increased two-fold. In wash-out experiments the hearts were pre-loaded with Rb+ for 30 min following which Rb+ wash-out was initiated. Four minutes later, flow was either decreased in the absence or presence of 10 microm 2,4-dinitrophenol (DNP), or 0.1 m m DNP was infused at normal flow for 20 min. LFI resulted in biphasic Rb+ efflux; during the initial phase, which lasted 8 min, the rate constant (kx10(3)/min) did not differ from control (43+/-3). The efflux was slightly inhibited by 5 microm glibenclamide (36+/-6) or 100 microm 5-hydroxydecanoic acid (32+/-4). In the second phase k decreased to half its initial value (18+/-2). More significant changes in energy state caused by LFI+10 microm DNP had no effect on the efflux kinetics. Similar changes in energy state induced by 0.1 m m DNP at normal flow were associated with activation of Rb+ efflux (71+/-5). DNP-stimulated Rb+ efflux was inhibited by acidosis (pHi approximately pHe = 6.7) produced with 5 m m morpholinoethane sulphonic acid (53+/-5) and by 100 microm adenosine (58+/-7). We suggest that accumulation of ischemic products such as H+and adenosine decreases activation of KATP channels in rat hearts., (Copyright 1999 Academic Press.)
- Published
- 1999
- Full Text
- View/download PDF
36. Measurements of mitochondrial K+ fluxes in whole rat hearts using 87Rb-NMR.
- Author
-
Gruwel ML, Kuzio B, Deslauriers R, and Kupriyanov VV
- Subjects
- 2,4-Dinitrophenol pharmacology, Animals, Cytosol metabolism, Heart drug effects, Heart physiology, Hydrogen-Ion Concentration, Hypothermia metabolism, In Vitro Techniques, Ion Exchange, Magnetic Resonance Spectroscopy, Male, Mitochondria, Heart drug effects, Phosphates metabolism, Rats, Rats, Sprague-Dawley, Rubidium metabolism, Rubidium Radioisotopes, Saponins pharmacology, Uncoupling Agents pharmacology, Mitochondria, Heart metabolism, Potassium metabolism
- Abstract
The rubidium efflux from hypothermic rat hearts perfused by the Langendorff method at 20 degreesC was studied. At this temperature 87Rb-NMR efflux experiments showed the existence of two 87Rb pools: cytoplasmic and mitochondrial. Rat heart mitochondria showed a very slow exchange of mitochondrial Rb+ for cytoplasmic K+. After washout of cytosolic Rb+, mitochondria kept a stable Rb+ level for >30 min. Rb+ efflux from mitochondria was stimulated with 0.1 mM 2, 4-dinitrophenol (DNP), by sarcolemmal permeabilization and concomitant cellular energy depletion by saponin (0.01 mg/ml for 4 min) in the presence of a perfusate mimicking intracellular conditions, or by ATP-sensitive K (KATP) channel openers. DNP, a mitochondrial uncoupler, caused the onset of mitochondrial Rb+ exchange; however, the washout was not complete (80 vs. 56% in control). Energy deprivation by saponin, which permeabilizes the sarcolemma, resulted in a rapid and complete Rb+ efflux. The mitochondrial Rb+ efflux rate constant (k) decreased in the presence of glibenclamide, a KATP channel inhibitor (5 microM; k = 0.204 +/- 0.065 min-1; n = 8), or in the presence of ATP plus phosphocreatine (1.0 and 5.0 mM, respectively; k = 0.134 +/- 0.021 min-1; n = 4) in the saponin experiments (saponin only; k = 0.321 +/- 0.079 min-1; n = 3), indicating the inhibition of mitochondrial KATP channels. Thus hypothermia in combination with 87Rb-NMR allowed the probing of the mitochondrial K+ pool in whole hearts without mitochondrial isolation.
- Published
- 1999
- Full Text
- View/download PDF
37. Temperature dependence of monovalent cation fluxes in isolated rat hearts: a magnetic resonance study.
- Author
-
Gruwel ML, Kuzio B, Xiang B, Deslauriers R, and Kupriyanov VV
- Subjects
- Animals, Cations, Monovalent, Cell Membrane Permeability, In Vitro Techniques, Ion Transport, Kinetics, Magnetic Resonance Spectroscopy, Male, Rats, Rats, Sprague-Dawley, Temperature, Lithium metabolism, Myocardium metabolism, Rubidium metabolism, Sodium metabolism
- Abstract
Ion flux studies were performed on Langendorff-perfused rat hearts using 87Rb, 7Li and 23Na NMR at 36, 20 and 10 degreesC, and at constant extracellular pH (7.40). Using 31P NMR, the intracellular pH was estimated and the high energy phosphate content monitored. Compared to 36 degreesC (k=0.044+/-0.015 min-1), our measurements showed incomplete Rb+ efflux with a dramatically (5-fold) increased rate constant, k, at 20 degreesC, k=0.238+/-0.080 min-1. 5 microM glibenclamide, a KATP-channel inhibitor, completely depressed the hypothermia-activated Rb+ efflux at this temperature (k=0.052+/-0. 018 min-1). 7Li NMR efflux studies on KCl-arrested hearts at 20 degreesC also showed an increase (3-fold) in efflux rate constant: k=0.090+/-0.003 min-1 relative to its value at 36 degreesC. At 10 degreesC, both Rb+ and Li+ showed efflux rate constants similar to those observed at 36 degreesC, k=0.071+/-0.016 min-1 and k=0.050+/-0. 005 min-1, respectively, and the washout was complete. 31P NMR at 36, 20 and 10 degreesC indicated cytosolic alkalinization at pH values of 7.05, 7.21 and 7.40, respectively. The ion transport data could be interpreted in terms of a myocyte model allowing for temperature-dependent changes in transport coefficients. The incomplete efflux of Rb+ at 20 degreesC may indicate the existence of a mitochondrial Rb+-pool with a very low Rb+ permeability for efflux. These findings correlate with previously observed membrane phase transitions in these systems.
- Published
- 1998
- Full Text
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38. Observation of two inorganic phosphate NMR resonances in the perfused hypothermic rat heart.
- Author
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Gruwel ML, Kuzio B, Deslauriers R, and Kupriyanov VV
- Subjects
- Animals, Hydrogen-Ion Concentration, In Vitro Techniques, Magnetic Resonance Spectroscopy, Male, Perfusion, Rats, Rats, Sprague-Dawley, Cold Temperature, Heart physiology, Myocardium metabolism, Organ Preservation, Phosphates metabolism
- Abstract
The effect of hypothermia on isolated perfused rat hearts was studied with 31P NMR. Hearts were continuously perfused with phosphate-free Krebs-Henseleit buffer while the perfusate temperature was adjusted. Perfusate pH was kept at 7.40 +/- 0.02 throughout the experiments. Using the chemical shift difference between PCr and Pi the intracellular pH was estimated. At 36, 20, and 10 degreesC a cytosolic alkalinization at a pH of 7.05 +/- 0.04, 7.21 +/- 0.05, and 7.40 +/- 0.03 was observed, respectively. At 10 degreesC two Pi resonances were observed with a separation of 0.25 ppm. This resonance corresponded to a Pi resonance of a cellular compartment with a local pH of 7.78 +/- 0.06, likely mitochondrial. This additional resonance disappeared upon warming of the hearts back to 36 degreesC.
- Published
- 1998
- Full Text
- View/download PDF
39. Three-dimensional 87Rb imaging of isolated pig hearts: effects of regional ischemia.
- Author
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Kupriyanov VV, Shen J, Xiang B, Kuzio B, Sun J, and Deslauriers R
- Subjects
- Animals, Myocardial Ischemia pathology, Necrosis, Phosphates metabolism, Potassium metabolism, Sodium-Potassium-Exchanging ATPase metabolism, Swine, Tissue Survival physiology, Image Processing, Computer-Assisted, Magnetic Resonance Imaging, Magnetic Resonance Spectroscopy, Myocardial Ischemia diagnosis, Rubidium Radioisotopes pharmacokinetics
- Abstract
The aim of this work was to investigate whether 1) Rb uptake is reduced in ischemic myocardium, and 2) 87Rb three-dimensional (3D) imaging can detect the ischemic area. Hearts of domestic pigs (n = 8, 20-30 kg) were perfused retrogradely with Krebs-Henseleit buffer in a 7-T, 40-cm horizontal bore magnet interfaced with Bruker MSLX spectrometer. Control (C) and ischemic (I) (45 min ligation of the left anterior descending coronary artery (LAD)) hearts were loaded with Rb+ by perfusion with a Rb(+)-containing solution (2-4.7 mM, 30-100% of K+ substitution) for 35 min and 87Rb (C) or 31P (ischemic area in I) spectra were acquired. After mechanical arrest with 0.6-0.9 mM lidocaine, 87Rb images (7 min each, 1 cm3 resolution) were acquired (30-40 min) in the presence of Rb(+)-containing perfusate. Subsequently, the hearts were stained with Evans blue (EB) and samples taken for measurements of Rb+ content. In the Group C, distribution of Rb+ in the left ventricle and the intensities of the 3D 87Rb images were uniform. In the ischemic area (Group I), verified by the lack of EB staining and changes in 31P spectra, the images showed a reduced intensity, which corresponded to decreased Rb+ content (33 +/- 11% of the normal). Thus, 87Rb imaging reveals damaged cells detecting reduced Rb+ content in the ischemic area.
- Published
- 1998
- Full Text
- View/download PDF
40. Effects of cromakalim and glibenclamide on myocardial high energy phosphates and intracellular pH during ischemia-reperfusion: 31P NMR studies.
- Author
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Docherty JC, Gunter HE, Kuzio B, Shoemaker L, Yang L, and Deslauriers R
- Subjects
- Animals, Anti-Arrhythmia Agents pharmacology, Coronary Circulation drug effects, Cromakalim, Decanoic Acids pharmacology, Dose-Response Relationship, Drug, Hydrogen-Ion Concentration, Hydroxy Acids pharmacology, Hypoglycemic Agents pharmacology, In Vitro Techniques, Lactates metabolism, Magnetic Resonance Spectroscopy, Male, Myocardial Ischemia metabolism, Myocardium metabolism, Phosphorus Isotopes, Rats, Rats, Sprague-Dawley, Adenosine Triphosphate metabolism, Benzopyrans pharmacology, Glyburide pharmacology, Myocardial Ischemia drug therapy, Myocardial Reperfusion, Pyrroles pharmacology
- Abstract
ATP sensitive potassium channel (KATP) openers (e.g. cromakalim) are thought to be cardioprotective during ischemia-reperfusion, while KATP blockers (e.g. glibenclamide) may potentiate ischemia-reperfusion damage. We studied cardiac energetics and intracellular pH, by 31P magnetic resonance spectroscopy, during ischemia-reperfusion of buffer perfused, isolated rat hearts in the presence of cromakalim (10 microM) or glibenclamide (1, 10 and 50 microM). Hearts were subjected to 25 min total global ischemia at 36.5 degrees C and reperfused for 45 min. Pre-treatment with cromakalim delayed the time to ischemic contracture (19.3 +/- 1.5 min v 15.3 +/- 0.6 for control, P < 0.05) and significantly improved recovery of function at 45 min reperfusion (84 +/- 11% pre-ischemic rate pressure product (RPP) v 38 +/- 5 for control, P < 0.05). This was accompanied by an attenuation in the loss of ATP during ischemia. Pre-treatment with glibenclamide decreased the time to ischemic contracture: 16.1 +/- 0.8 min. 15.1 +/- 0.7, 12.0 +/- 1.2 (P < 0.01) and 9.5 +/- 0.9 (P < 0.001) for control, 1, 10 and 50 microM glibenclamide respectively. 50 microM glibenclamide significantly improved functional recovery at 45 min reperfusion but 1 and 10 microM were without effect; 24 +/- 6, 22 +/- 4, 29 +/- 4 and 58 +/- 7% (P < 0.05) of pre-ischemic RPP for control, 1, 10 and 50 microM glibenclamide. During ischemia, intracellular ATP was depleted more rapidly in the presence of 50 microM glibenclamide and intracellular acidosis was significantly attenuated (final pH 6.3 v 5.8 for control). 50 microM glibenclamide also decreased tissue lactate content at the end of ischemia (75 +/- 3 mumol/g dry weight v 125 +/- 18 for control, P < 0.05) and this attenuation of lactate accumulation and consequent decreased intracellular acidosis may be responsible for the cardioprotection observed under these conditions. These latter effects are unlikely to be related to glibenclamide's KATP blocking activity. This study demonstrates that blocking of myocardial KATP does not potentiate ischemia-reperfusion injury and, in addition, illustrates the important role played by intracellular acidosis in myocardial ischemia-reperfusion injury.
- Published
- 1997
- Full Text
- View/download PDF
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