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OBJECTIVE: The pair-wise addition of parahydrogen, the singlet form of molecular hydrogen, to unsaturated precursors evokes the hyperpolarization of two parahydrogen-derived 1H nuclear spins through a process known as parahydrogen-induced polarization (PHIP). Subsequent spin order transfer (SOT) from the 1H to the surrounding 13C nuclear spins via magnetic field cycling (MFC) results in substantial signal enhancement in 13C magnetic resonance imaging (MRI). Here, we report the development of a unique PHIP 13C hyperpolarizer system using a flow guide for MFC. METHODS: The optimal MFC scheme for 1H to 13C spin order transfer was quantum-chemically simulated using the J-coupling values of 13C-labeled metabolic tracers. The flow guide system was three-dimensionally designed based on the simulated MFC scheme and pre-measured magnetic field distribution in a zero-field chamber. RESULTS: The system efficiently transfers the spin order of hyperpolarized 1H to a particular 13C spin when the parahydrogenated tracer passes through the flow guide at a designated flow rate. The 13C MRI signal is enhanced more than 40,000 times in 13C-labeled pyruvate and fumarate, compared to the thermal equilibrium level at 1.5 T, was achieved for conducting in vivo metabolic MRI of mice. CONCLUSION: A fully automated PHIP-based 13C polarizer was developed using a unique flow guide to conduct the MFC for 1H to 13C SOT. SIGNIFICANCE: The PHIP hyperpolarizer with a flow guide can conduct efficient 1H-13C SOT without a MFC magnetic field sweep system and offers a cost-effective alternative to conventional dynamic nuclear polarization.
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Isótopos de Carbono , Hidrogênio , Campos Magnéticos , Imageamento por Ressonância Magnética , Animais , Camundongos , Hidrogênio/química , Imageamento por Ressonância Magnética/métodos , Isótopos de Carbono/química , Imagens de Fantasmas , Desenho de EquipamentoRESUMO
Significance:In vivo molecular and metabolic imaging is an emerging field in biomedical research that aims to perform noninvasive detection of tissue metabolism in disease states and responses to therapeutic agents. The imbalance in tissue oxidation/reduction (Redox) states is related to the onset and progression of several diseases. Tissue redox metabolism provides biomarkers for early diagnosis and drug treatments. Thus, noninvasive imaging of redox metabolism could be a useful, novel diagnostic tool for diagnosis of redox-related disease and drug discovery. Recent Advances:In vivo dynamic nuclear polarization magnetic resonance imaging (DNP-MRI) is a technique that enables the imaging of free radicals in living animals. DNP enhances the MRI signal by irradiating the target tissue or solution with the free radical molecule's electron paramagnetic resonance frequency before executing pulse sequence of the MRI. In vivo DNP-MRI with redox-sensitive nitroxyl radicals as the DNP redox contrast agent enables the imaging of the redox metabolism on various diseases. Moreover, nitroxyl radicals show antioxidant effects that suppress oxidative stress. Critical Issues: To date, considerable progress has been documented preclinically in the development of animal imaging systems. Here, we review redox imaging of in vivo DNP-MRI with a focus on the recent progress of this system and its uses in patients with redox-related diseases. Future Directions: This technique could have broad applications in the study of other redox-related diseases, such as cancer, inflammation, and neurological disorders, and facilitate the evaluation of treatment response as a theranostic tool. Antioxid. Redox Signal. 36, 172-184.
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Imageamento por Ressonância Magnética , Medicina de Precisão , Animais , Espectroscopia de Ressonância de Spin Eletrônica/métodos , Radicais Livres , Humanos , Imageamento por Ressonância Magnética/métodos , OxirreduçãoRESUMO
Free radicals, such as metabolic intermediates, reactive oxygen species, and metal enzymes, are key substances in organisms, although they can also cause various oxidative diseases. Thus, in vivo free radical imaging should be considered as the ultimate form of metabolic imaging. Unfortunately, electron spin resonance (ESR) imaging has inherent disadvantages, such as free radicals with large linewidths generating blurred images and the presence of two or more free radicals resulting in a complicated imaging procedure. Dynamic nuclear polarization-magnetic resonance imaging (DNP-MRI) is a noninvasive imaging method to visualize in vivo free radicals, theoretically, with the same resolution as the MRI anatomical resolution, and fixed low-field DNP-MRI provides unique information on oxidative diseases and cancer. However, the large gyromagnetic ratio of the electron spin, which is 660-fold greater than that of a proton, requires field cycling, wherein the external magnetic field should be varied during DNP-MRI observations. This causes difficulties in developing a DNP-MRI system for clinical purposes. We developed a novel field-cycling DNP-MRI system for a preclinical study. In the said system, the magnetic field is switched by rotationally moving two magnets, with a magnetic flux density of 0.3 T for MRI and 5 mT for ESR. The image quality was examined using various pulse sequences and ESR irradiation using nitroxyl radical as the phantom, and the optimum conditions were established. Using the system, we performed a preclinical study involving free radical imaging by placing the free radicals under the palm of a human hand.
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Imageamento por Ressonância Magnética , Espectroscopia de Ressonância de Spin Eletrônica , Radicais Livres , Humanos , Oxirredução , Imagens de FantasmasRESUMO
The front cover artwork is provided by the group of Dr. Neil J. Stewart, Prof. Hiroshi Hirata, and Dr. Shingo Matsumoto (Hokkaido University, Japan) as well as Dr. Takuya Hashimoto (Chiba University, Japan). The image shows hyperpolarized 13 C fumarate metabolism to hyperpolarized 13 C malate, which is released into the extracellular space in regions of necrotic cell death, where the cell membrane is disrupted. Read the full text of the Article at 10.1002/cphc.202001038.
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Hyperpolarized [1-13 C]fumarate is a promising magnetic resonance imaging (MRI) biomarker for cellular necrosis, which plays an important role in various disease and cancerous pathological processes. To demonstrate the feasibility of MRI of [1-13 C]fumarate metabolism using parahydrogen-induced polarization (PHIP), a low-cost alternative to dissolution dynamic nuclear polarization (dDNP), a cost-effective and high-yield synthetic pathway of hydrogenation precursor [1-13 C]acetylenedicarboxylate (ADC) was developed. The trans-selectivity of the hydrogenation reaction of ADC using a ruthenium-based catalyst was elucidated employing density functional theory (DFT) simulations. A simple PHIP set-up was used to generate hyperpolarized [1-13 C]fumarate at sufficient 13 C polarization for exâ vivo detection of hyperpolarized 13 C malate metabolized from fumarate in murine liver tissue homogenates, and inâ vivo 13 C MR spectroscopy and imaging in a murine model of acetaminophen-induced hepatitis.
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Ácidos Graxos Insaturados/biossíntese , Fumaratos/metabolismo , Imageamento por Ressonância Magnética , Alcinos/química , Isótopos de Carbono , Teoria da Densidade Funcional , Ácidos Graxos Insaturados/química , Fumaratos/química , HidrogenaçãoRESUMO
Reactive oxygen species (ROS) play an important role in cell metabolism, but they can cause oxidative damage to biomolecules. Among ROS, the hydroxyl radical (·OH) is one of the most reactive molecules in biological systems because of its high reaction rate constant. Therefore, imaging of ·OH could be useful for evaluation of the redox mechanism and diagnosis of oxidative diseases. In vivo dynamic nuclear polarization-magnetic resonance imaging (DNP-MRI) is a noninvasive imaging method to obtain spatiotemporal information about free radicals with MRI anatomical resolution. In this study, we investigated the visualization of hydroxyl radicals generated from the Fenton reaction by combining DNP-MRI with a spin-trapping agent (DMPO: 5,5-dimethyl-1-pyrroline N-oxide) for ·OH. Additionally, we demonstrated the radical-scavenging effect using four thiol-related reagents by DNP-MRI. We demonstrated that DNP enhancement could be induced by the DMPO-OH radical using the DNP-MRI/spin-trapping method and visualized ·OH generation for the first time. Maximum DNP enhancement was observed at an electron paramagnetic resonance irradiation frequency of 474.5 MHz. Furthermore, the radical-scavenging effect was simultaneously evaluated by the decrease in the DNP image value of DMPO-OH. An advantage of our methods is that they simultaneously investigate compound activity and the radical-scavenging effect.
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Radical Hidroxila/química , Radical Hidroxila/metabolismo , Imageamento por Ressonância Magnética , Detecção de Spin , Óxidos N-Cíclicos/química , Peróxido de Hidrogênio/química , Ferro/química , Fatores de TempoRESUMO
For the construction of metal-free magnetic resonance imaging (MRI) contrast agents, radical-based nanoparticles (RNPs) are promising materials because they allow the water-proton longitudinal relaxivity (r1) to be enhanced not only by paramagnetic resonance effects but also by prolonging the rotational correlation times (τR). However, the τR effect is limited because the radical units are often located within the central hydrophobic core of oil-in-water (o/w) emulsions, resulting in a lack of water molecules surrounding the radical units. In this study, to construct supramolecular RNPs that have high r1 values, we designed a liposome-type RNP in which the radical units are located at positions with sufficient surrounding water molecules. Using this strategy, PRO1 with a PROXYL framework was prepared by introducing hydrophilic groups on both sides of the radical unit. The RNP composed of PRO1 formed spherical nanoparticles approximately 100 nm in size and yielded a higher r1 value (0.26 mM-1 s-1) compared to those of small radical species and similar supramolecular o/w emulsion-type nanoparticles (0.17 mM-1 s-1 in PRO2).
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Materials possessing electron spin can shorten the T 1 relaxation times in magnetic resonance imaging (MRI). For example, gadolinium (Gd) complexes with seven f-orbital electrons are widely used as contrast agents in clinical applications. However, Gd has severe potential side effects, and thus metal-free alternatives are needed. Toward this end, we synthesized seven NO radicals consisting of a dioxa-azaspiro[4.5]decane framework having various substituents, DAD-X (X = methyl, ethyl, n-propyl, c-propyl, vinyl, phenyl, and 2-pyridyl), that functioned as metal-free MRI contrast agents. The relationship between (i) water-proton relaxivity and logâ¯P and (ii) reactivity for ascorbic acid and the spin density of the NO oxygen atom were established, which provided a basis for the rational design of practical metal-free contrast agents.
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Melanin is a pigment that includes free radicals and is widely distributed in living animals. Malignant melanoma is one of the most progressive tumors in humans with increasing incidence worldwide, and has shown resistance to chemotherapy, resulting in high mortality at the metastatic stage. In general, melanoma involves the abnormal accumulation of melanin pigment produced by malignant melanocytes. Electron paramagnetic resonance (EPR) spectroscopy and imaging is a powerful technique to directly visualize melanomas using endogenous free radicals in the melanin pigment. Because melanin radicals have a large linewidth, the low spatial resolution of EPR imaging results in blurred images and a lack of anatomical information. Dynamic nuclear polarization (DNP)-MRI is a noninvasive imaging method to obtain the spatio-temporal information of free radicals with MRI anatomical resolution. Proton signals in tissues, including free radicals, can be dramatically enhanced by EPR irradiation at the resonance frequency of the free radical prior to applying the MRI pulse sequence. However, the DNP effects of free radicals in the pigment of living organisms is unclear. Therefore, if endogenous free radicals in melanin pigment could be utilized as a bio-probe for DNP-MRI, this will be an advantage for the specific enhancement of melanoma tissues and might allow the separate noninvasive visualization of melanoma tissues without the need for probe administration. Here, we report that biological melanin pigment induced a in vivo DNP effect by interacting with water molecules. In addition, we demonstrated in vivo melanoma imaging based on the DNP effects of endogenous free radicals in the melanin pigment of living mice.
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Imageamento por Ressonância Magnética/métodos , Melaninas/metabolismo , Melanoma Experimental/patologia , Ressonância Magnética Nuclear Biomolecular/métodos , Processamento de Sinais Assistido por Computador , Animais , Feminino , Melanoma Experimental/metabolismo , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Overhauser enhanced MRI (OMRI) is one of the free radical imaging technologies and has been used in biomedical research such as for partial oxygen measurements in tumor, and redox status in acute oxidative diseases. The external magnetic field of OMRI is frequently in the range of 5-10 mTesla to ensure microwave penetration into small animals, and the S/N ratio is limited. In this study, a 0.15 Tesla OMRI was constructed and tested to improve the S/N ratio for a small sample, or skin measurement. Specification of the main magnet was as follows: 0.15 Tesla permanent magnet; gap size 160 mm; homogenous spherical volume of 80 mm in diameter. The OMRI resonator was designed based on TE101 cavity mode and machined from a phosphorus deoxidized copper block for electron spin resonance (ESR) excitation and a solenoid transmission/receive resonator for NMR detection. The resonant frequencies and Q values were 6.38 MHz/150 and 4.31-4.41 GHz/120 for NMR and ESR, respectively. The Q values were comparable to those of conventional low field OMRI resonators at 15 mTesla. As expected, the MRI S/N ratio was improved by a factor of 30. Triplet dynamic nuclear polarization spectra were observed for 14N carboxy-PROXYL, along the excitation microwave sweep. In the current setup, the enhancement factor was ca. 0.5. In conclusion, the results of this preliminary evaluation indicate that the 0.15 Tesla OMRI could be useful for free radical measurement for small samples.
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Aumento da Imagem , Imageamento por Ressonância Magnética/instrumentação , Imageamento por Ressonância Magnética/métodos , Imãs/química , Espectroscopia de Ressonância de Spin Eletrônica , Radicais Livres , Aumento da Imagem/instrumentação , Aumento da Imagem/métodos , Espectroscopia de Ressonância Magnética , Imagem Molecular/instrumentação , Imagem Molecular/métodos , Oxirredução , Oxigênio/análise , Oxigênio/metabolismo , Pressão ParcialRESUMO
In hyperpolarization of (13)C-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 (13)C nuclear spins. Here, we report systematic measurement of the spin-lattice relaxation time (T1) and enhancement level of (13)C-enriched chemical probes in the presence of various Gd contrast agents in the liquid-state after dissolution. Using two different (13)C probes having opposite electric charges at neutral pH, we clearly show the T1 of hyperpolarized (13)C was barely affected by the use of a Gd complex that displays repulsive interaction with the (13)C probe in solution, whilst T1 was drastically shortened when there was ionic attraction between probe and complex.
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For indirect tissue observation, electron-spin, Overhauser-enhanced, dynamic nuclear polarization magnetic resonance imaging (DNP-MRI) is a useful technique. However, its sensitivity and resolution are low compared with the clinical MRI apparatus. By switching to electron spin resonance (ESR) excitation, the magnetic field of the NMR detection, field cycle technique, which aims to improve resolution, was proposed. However, the effect of eddy currents or current value was altered unsatisfactorily. A team at Kyushu University proposed a new DNP-MRI technique capable of improving NMR detection field by preparing in advance a magnetic field, which was connected by the sample transport system. By developing a mobile MRI method that can be used while moving, and fastening the sample in a disk that rotates at a constant speed, they have developed a circular transport DNP-MRI method that greatly reduces the load on the sample. The circular transport DNP-MRI system comprises a circular sample transport system, detection of an MRI magnetic field of 1.5 T, and ESR excitation magnetic field of 20 mT. The developed DNP-MRI had a clear glass tube phantom and resolution of 0.15 mm, and was successful in imaging multiple radical resonant points. It has been commercialized by Japan Redox Limited. In the process of equipment commercialization, a new digital spectrometer has been developed, which expanded the MRI apparatus.
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Imagem Molecular/instrumentação , Imagem Molecular/tendências , Oxirredução , Animais , Espectroscopia de Ressonância de Spin Eletrônica , Humanos , Imageamento por Ressonância Magnética , Espectroscopia de Ressonância Magnética , Imagem Molecular/métodosRESUMO
Overhauser-enhanced magnetic resonance imaging (OMRI), which is a double resonance technique, creates images of free radical distribution in animals by enhancing the water proton signal intensity by the overhauser effect. In this study, we constructed a contrast agent by combining PROXYL groups that have nitroxyl radicals with PEG-modified dendritic poly(l-lysine) that accumulates in the tumor by enhanced permeability and retention (EPR) effect. Addition of the PROXYL groups at the PEG chains' termini on KG6 was advantageous in OMRI, because the ESR signal of the nitroxyl radical was maintained without decay caused by mobility restriction, even if the PROXYL groups were attached at 25 mol% on one molecule. After intramuscular injection of the molecule modified at 25 mol%, that is, PR25-PEG-KG6, a significant OMRI signal was observed at the injected site. However, no signal was detected in the tumor after intravenous injection of PR25-PEG-KG6 to a tumor-bearing mouse, although PR25-PEG-KG6 itself accumulated in the tumor. The reason was that the nitroxyl radicals were immediately reduced in the blood after the injection, suggesting that use of stable nitroxyl radicals will enable detection of tumors by OMRI after intravenous injection.
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Meios de Contraste , Dendrímeros , Imageamento por Ressonância Magnética/métodos , Neoplasias Experimentais/patologia , Polietilenoglicóis , Polilisina , Animais , Neoplasias da Mama/patologia , Carcinoma/patologia , Linhagem Celular Tumoral , Meios de Contraste/síntese química , Meios de Contraste/química , Dendrímeros/síntese química , Dendrímeros/química , Radicais Livres/química , Masculino , Camundongos , Imagem Óptica , Polietilenoglicóis/síntese química , Polietilenoglicóis/química , Polilisina/síntese química , Polilisina/químicaRESUMO
Overhauser-enhanced MRI (OMRI) enables visualization of free radicals in animals based on dynamic nuclear polarization. Real-time data of tissue redox status gathered from kinetic images of redox-sensitive nitroxyl radical probes using OMRI provided both anatomic and physiological information. Phantom experiments demonstrated the linear correlation between the enhancement factor and the concentration of a membrane-impermeable probe, carboxy-PROXYL (3-carboxy-2,2,5,5-tetramethyl- pyrrolidine-1-oxyl). Whole-body OMRI images illustrated the in vivo kinetics of carboxy-PROXYL for 25 min. Initial distribution was observed in lung, heart, liver, and kidney, but not brain, corresponding to its minimal lipophilicity. Based on these images (pixel size, 1.33 × 1.33 mm; slice thickness, 50mm), a time-concentration curve with low coefficient of variance (<0.21) was created to assess pharmacokinetic behaviors. A biexponential curve showed a distribution phase from 1 to 10 min and an elimination phase from 15 to 25 min. The α rate constant was greater than the ß rate constant in ROIs, confirming that its pharmacokinetics obeyed a two-compartment model. As a noninvasive technique, combining OMRI imaging with redox probes to monitor tissue redox status may be useful in acquiring valuable information regarding organ function for preclinical and clinical studies of oxidative diseases.
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Óxidos N-Cíclicos/farmacocinética , Imageamento por Ressonância Magnética , Sondas Moleculares/farmacocinética , Óxidos de Nitrogênio/análise , Imagens de Fantasmas , Pirrolidinas/farmacocinética , Animais , Feminino , Processamento de Imagem Assistida por Computador , Cinética , Imageamento por Ressonância Magnética/instrumentação , Imageamento por Ressonância Magnética/métodos , Camundongos , Oxirredução , Marcadores de Spin , Fatores de TempoRESUMO
Nonsteroidal anti-inflammatory drugs are the drugs of choice in the treatment of rheumatoid arthritis (RA) because of their rapid analgesic effect. However, they induce severe gastric damage in RA patients and animals by a process mediated by reactive oxygen species (ROS). Nitroxides (nitroxyl radicals) are widely used as imaging agents and antioxidants to explore the role of ROS generation in the pathogenesis of disease. In this study, the effectiveness of the newly synthesized nitroxides 8-aza-7,7,9,9-tetramethyl-1,4-dioxaspiro[4.5]undecan-8-oxyl (compound 1) and 4-oxo-2,2,6,6-tetraethylpiperidine-1-oxyl (compound 2) in the prevention of gastric ulcers in adjuvant arthritis rats treated with indomethacin was evaluated by monitoring the reaction of reactive oxygen species in gastric tissue with Overhauser-enhanced magnetic resonance imaging (OMRI). Pretreatment with all tested nitroxides suppressed the ulcers induced by indomethacin treatment in arthritic rats. OMRI using compounds 1 and 2 as well as 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL) demonstrated a redox imbalance in the stomach of these rats. Lipid peroxide and interleukin (IL)-1ß levels in the gastric mucosa were significantly suppressed by compound 1 and TEMPOL, whereas CINC/gro, a member of the IL-8 family, was significantly suppressed by compound 1 only. These results suggest that the preventive effects of nitroxides on gastric ulcers may operate by different mechanisms.
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Artrite Experimental/metabolismo , Óxidos de Nitrogênio/metabolismo , Úlcera Gástrica/prevenção & controle , Animais , Anti-Inflamatórios não Esteroides/efeitos adversos , Antioxidantes/metabolismo , Artrite Experimental/tratamento farmacológico , Artrite Experimental/patologia , Indometacina/efeitos adversos , Imageamento por Ressonância Magnética , Masculino , Ratos , Ratos Endogâmicos , Espécies Reativas de Oxigênio/metabolismo , Úlcera Gástrica/induzido quimicamente , Úlcera Gástrica/metabolismoRESUMO
Oxidative stress may be involved in the dopaminergic neurodegenerations seen in 6-OHDA-lesioned rats through its production of free radicals and through mitochondrial dysfunction. In this study, we noninvasively demonstrate brain redox alterations in 6-OHDA-lesioned rats using Overhauser-enhanced magnetic resonance imaging (OMRI). The reduction rate of 3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine-l-oxyl (methoxycarbonyl-PROXYL), a redox-sensitive contrast agent, was used as an index of the redox status in vivo. The methoxycarbonyl-PROXYL reduction rate, calculated from continuous images, decreased significantly in lesioned hemispheres compared to their corresponding contralateral hemispheres. The reduction rates in cellular fractions obtained from the striatum were estimated by X-band electron spin resonance (ESR) and calculated by assuming first-order kinetics for their time-dependent decreases. When methoxycarbonyl-PROXYL was mixed with cytoplasm fractions, the reduction rates were the same in both hemispheres. However, the ESR signal of methoxycarbonyl-PROXYL in the mitochondrial fraction of the lesioned hemispheres decayed more slowly than that of the corresponding contralateral hemispheres. Concordantly, biochemical assays showed that the activity of mitochondrial complex I also decreased more slowly in lesioned hemispheres. Thus, this method of noninvasively imaging brain redox alterations faithfully reflects changes in mitochondrial complex I activity in 6-OHDA-lesioned rats.
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Corpo Estriado/metabolismo , Modelos Animais de Doenças , Metabolismo Energético/fisiologia , Imageamento por Ressonância Magnética/métodos , Mitocôndrias/patologia , Estresse Oxidativo/fisiologia , Oxidopamina/administração & dosagem , Transtornos Parkinsonianos/metabolismo , Transtornos Parkinsonianos/patologia , Animais , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/fisiopatologia , Metabolismo Energético/efeitos dos fármacos , Masculino , Mitocôndrias/fisiologia , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Transtornos Parkinsonianos/induzido quimicamente , Ratos , Ratos Sprague-Dawley , Frações SubcelularesRESUMO
Overhauser-enhanced magnetic resonance imaging (OMRI) studies of a membrane-permeable nitroxyl spin probe, (2)H-enriched 3-methoxycarbonyl-2,2,5,5-tetramethyl-pyrrolidine-1-oxyl (MC-PROXYL), used in simultaneous molecular imaging is reported. Phantom imaging was performed with liposomal solutions of MC-PROXYL at varying spin probe and liposome concentrations using a field-cycle mode, custom-built OMRI scanner. Dynamic nuclear polarization (DNP) spectra of the liposomal solution of the spin probe, measured at 14.529mT using a 5mT sweep of the electron paramagnetic resonance (EPR) irradiation field showed splitting of the low and high filed hyperfine lines. Spectral measurements using D(2)O and a spin broadening agent, K(3)Fe(CN)(6) confirmed that these peaks originated from water molecules in two different environments, compartmentalized with liposomes. The nuclear Overhauser enhancement measured at different EPR irradiation times and power levels showed reduction in water nuclear magnetic resonance (NMR) signal enhancement in liposomal membrane due to the reduction in the coupling constant, rho. This study illustrates that OMRI can be used to differentiate between the intra- and extra- membrane water by loading the liposome vesicles with a lipid-permeable nitroxyl spin probe.
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Espectroscopia de Ressonância de Spin Eletrônica/métodos , Lipossomos/química , Técnicas de Sonda Molecular , Óxidos de Nitrogênio/análise , Óxidos de Nitrogênio/química , Água/química , Soluções , Marcadores de SpinRESUMO
Reactive oxygen species (ROS) are thought to play a significant role in the development of diabetic retinopathy; however, no direct evidence supports ROS generation in vivo. This study used in vivo electron spin resonance (ESR) spectroscopy with a surface resonator to detect local free radical reactions. The ESR signal decay of carbamoyl-PROXYL was enhanced in the eyes of streptozotocin (STZ)-induced diabetic mice. This enhanced signal decay was suppressed by the administration of SOD or the pretreatment with aminoguanidine. We demonstrate, for the first time, specific free radical reactions in the eyes of mice with STZ-induced diabetes.