Dynamic Nuclear Polarization-magnetic Resonance Imaging Research Articles

Non-invasive differentiation of hepatic steatosis and steatohepatitis in a mouse model using nitroxyl radical as an MRI-contrast agent

Drug-induced steatohepatitis is considered more serious than drug-induced hepatic steatosis, so that differentiating between the two is crucial in drug development. In addition, early detection of drug-induced steatohepatitis is considered important since recovery is possible with drug withdrawal. However, no method has been established to differentiate between the two. In the development of drug-induced steatohepatitis, reactive oxygen species (ROS) is excessively generated in the liver. It has been reported that ROS can be monitored with electron spin resonance (ESR) and dynamic nuclear polarization-magnetic resonance imaging (DNP-MRI) by using nitroxyl radicals, which are known to participate in various in vivo redox reactions. The decay/reduction rate, which is an index for monitoring nitroxyl radicals, has been reported to be increased in tissues with excessive ROS levels other than liver, but decreased in methionine choline deficient (MCD) diet-induced steatohepatitis with excess ROS. Therefore, looking to differentiate between drug-induced hepatic steatosis and steatohepatitis, we examined whether the reduction rate decreases in steatohepatitis other than the MCD-diet induced disease and whether the decrease could be detected by MRI. We used STAM™ mice in which hepatic steatosis and steatohepatitis developed sequentially under diabetic conditions. 3-carbamoyl-PROXYL (CmP), one of the nitroxyl radicals, was injected intravenously during the MRI procedure and the reduction rate was calculated. The reduction rate was significantly higher in early steatohepatitis than in hepatic steatosis and the control. Excess ROS in early steatohepatitis was detected by an immunohistochemical marker for ROS. Therefore, it was indicated that the increase or decrease in the reduction rate in steatohepatitis differs depending on the model, and early steatohepatitis could be noninvasively differentiated from hepatic steatosis using CmP in MRI. Since the change in direction of the reduction rate in steatohepatitis in clinical studies could be predicted by confirming the reduction rate in preclinical studies, the present method, which can be used consistently in clinical and preclinical studies, warrants consideration as a candidate monitoring method for differentiating between early drug-induced steatohepatitis and hepatic steatosis in drug development.

Noninvasive Diagnosis of the Mitochondrial Function of Doxorubicin-Induced Cardiomyopathy Using In Vivo Dynamic Nuclear Polarization-Magnetic Resonance Imaging.

Doxorubicin (DOX) induces dose-dependent cardiotoxicity via oxidative stress and abnormal mitochondrial function in the myocardium. Therefore, a noninvasive in vivo imaging procedure for monitoring the redox status of the heart may aid in monitoring diseases and developing treatments. However, an appropriate technique has yet to be developed. In this study, we demonstrate a technique for detecting and visualizing the redox status of the heart using in vivo dynamic nuclear polarization–magnetic resonance imaging (DNP–MRI) with 3-carbamoyl-PROXYL (CmP) as a molecular imaging probe. Male C57BL/6N mice were administered DOX (20 mg/kg) or saline. DNP–MRI clearly showed a slower DNP signal reduction in the DOX group than in the control group. Importantly, the difference in the DNP signal reduction rate between the two groups occurred earlier than that detected by physiological examination or clinical symptoms. In an in vitro experiment, KCN (an inhibitor of complex IV in the mitochondrial electron transport chain) and DOX inhibited the electron paramagnetic resonance change in H9c2 cardiomyocytes, suggesting that the redox metabolism of CmP in the myocardium is mitochondrion-dependent. Therefore, this molecular imaging technique has the potential to monitor the dynamics of redox metabolic changes in DOX-induced cardiomyopathy and facilitate an early diagnosis of this condition.

Development and Preclinical Study of Free Radical Imaging Using Field-Cycling Dynamic Nuclear Polarization MRI.

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.

In Vivo Dynamic Nuclear Polarization Magnetic Resonance Imaging for the Evaluation of Redox-Related Diseases and Theranostics.

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.

Imaging of Hydroxyl-Radical Generation Using Dynamic Nuclear Polarization-Magnetic Resonance Imaging and a Spin-Trapping Agent.

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.

Free Radical Imaging for Visualization of Radiation Treatment Using Dynamic Nuclear Polarization (DNP) MRI

In vivo dynamic nuclear polarization-magnetic resonance imaging (DNP-MRI) enables the anatomical distribution of free radical species to be monitored. DNP enhances the MRI signal from nuclei such as 1H by irradiating tissue at the electron paramagnetic resonance (EPR) frequency of the free radical, before applying the MRI pulse sequence, thus increasing the image intensity where free radicals are present. Low-molecular-weight, stable nitroxyl compounds, including Tempol and carbamoyl-PROXYL (CmP), are organic free radicals that show antioxidant capability, and they have been used as redox sensitive contrast agent. The reduction of the nitroxyl compounds depend on production of free radicals which may be influenced by factors such as overproduction of reactive oxygen species (ROS) after radiation treatment. In this study, we evaluated the feasibility of visualization of free radical generation in solution and redox alteration of mice liver induced by radiation treatment, assessed by DNP-MRI with Tempol and CmP as a molecular imaging probe. Free radical imaging for monitoring of redox reaction was performed with a low magnetic field using an in vivo DNP-MRI system (Keller, Japan Redox Inc.). DNP-MRI image of phantom including Tempol or CmP and glutathion (GSH) as reductants was obtained before and after radiation treatment (0, 5, 10, 20 Gy). Quantification of Tempol and CmP radical was performed by electron paramagnetic resonance (EPR) spectroscopy. Pharmacokinetic DNP-MRI images of liver homogenate or mitochondrial solution with or without radiation (10 or 20 Gy) were obtained at 3, 5, 10, 15, 20, 25, 30, 35 and 40 min after addition of redox probes (1mM Tempol or CmP). The redox map was obtained by the slope of the enhanced DNP image intensity of each pixel from four pharmacokinetic images using a custom Excel macro program. Direct redox reaction between 0.3mM Tempol and 10 mM GSH was observed by EPR experiments. Therefore, we determined that 1mM GSH is suitable concentration for monitoring the redox reaction by radiation treatment. DNP-MRI clearly visualized the reduction of Tempol free radical. The DNP images clearly showed the significant enhancement in tissue homogenate and mitochondrial solution. The enhancement of DNP image was gradually reduced by liver homogenate solution due to reduction of Tempol and the reduction rate of Tempol was increased by the treatment of radiation. Interestingly, reduction of Tempol in both homogenates with or without radiation were completely inhibited by KCN treatment. It is suggested that enhancement of Tempol reduction is related in mitochondrial electron transfer system. We demonstrated that the visualization of ROS production by radiation treatment and mitochondrial redox metabolic change in the electron transfer chain using DNP-MRI.

Dynamic nuclear polarization magnetic resonance imaging and the oxygen-sensitive paramagnetic agent OX63 provide a noninvasive quantitative evaluation of kidney hypoxia in diabetic mice

Renal hypoxia may play an important role in the progression of diabetic nephropathy. However, tools that noninvasively and quantitatively measure oxygen tension in the kidney are lacking. Here, we evaluated the feasibility of a noninvasive and quantitative imaging technique using dynamic nuclear polarization magnetic resonance imaging (DNP-MRI) in combination with the oxygen-sensitive paramagnetic agent OX63 for measuring oxygen tension in the kidney. Our results demonstrate that the DNP-MRI technique can yield quantitative maps of oxygen tension in the mouse renal cortex. Using this procedure, we also showed that oxygen tension was less elevated in the renal cortex of both streptozotocin-induced type 1 diabetic mice and db/db mice, a model of type 2 diabetes, than in the renal cortex of age-matched control mice of each respective model. Oxygen tension in streptozotocin-exposed mice was significantly improved by insulin treatment. Thus, the noninvasive and quantitative DNP-MRI technique appears to be useful for studying the pathophysiological role of hypoxia.

192 - Redox metabolic imaging of NASH model mice using in vivo dynamic nuclear polarization MRI

As a consequence of the global epidemic of metabolic syndrome and chronic obesity, non-alcoholic fatty liver disease (NAFLD) has emerged as an important contribution in several chronic liver diseases. The development of a non-invasive diagnostic method for assessing disease progression from NAFLD to non-alcoholic steatohepatitis (NASH) has become an important research goal. Oxidative stress is thought to play a central role in the progression of NAFLD to NASH. Mitochondria are involved in both ROS generation, and free fatty acid (FFA) s-oxidation that promotes lipid accumulation. Currently available non-invasive imaging technologies are only able to assess fat accumulation in the liver. Therefore, these methods are not suitable for a precise diagnosis of NASH. The standard diagnostic technique for NASH, liver biopsy, has several drawbacks, including the higher risk of complications that accompanies invasive procedures. Here, we demonstrated that in vivo mitochondrial redox metabolism was dramatically altered at an early stage, before histopathological changes, and NASH could be accurately diagnosed by in vivo dynamic nuclear polarization-magnetic resonance imaging in methionine-choline-deficient (MCD) diet-induced NASH mouse model. In addition, this technique was feasible for the diagnosis of NASH compared with histopathological findings from biopsies. Our data reveal a novel method for monitoring the dynamics of redox metabolic changes in NAFLD/NASH.

Imaging of Mitochondria Metabolic Change after Radiation Treatment using Dynamic Nuclear Polarization (DNP)-MRI

In vivo dynamic nuclear polarization-magnetic resonance imaging (DNP-MRI) enables the anatomical distribution of free radical species to be monitored. DNP enhances the MRI signal from nuclei such as 1H by irradiating tissue at the electron paramagnetic resonance (EPR) frequency of the free radical, before applying the MRI pulse sequence, thus increasing the image intensity where free radicals are present. Low-molecular-weight, stable nitroxyl compounds, including carbamoyl-PROXYL (CmP), are organic free radicals that possess SOD-like activity, and they have been used as probes in a variety of biophysical and biochemical experiments. The reduction rate of the radical probe depends on the tissue redox environment, which is influenced by factors such as overproduction of ROS and decreased antioxidant defences. In this study, we evaluated the feasibility of mitochondrial redox metabolic imaging, assessed by DNP-MRI with CmP as a molecular imaging probe, for detecting the metabolic change after radiation treatment to the tissue homogenate solution. Redox imaging of mitochondrial metabolism was performed with a low magnetic field using an in vivo DNP-MRI system (Keller, Japan Redox Inc.). The external magnetic field (B0) for EPR irradiation and MRI was fixed at 15 mT, and the radiofrequencies for EPR irradiation and MRI were 455 MHz and 683 kHz, respectively. DNP-MRI images of liver homogenate solution with or without radiation (10 or 20 gray) were obtained at 3, 5, 10, 15, 20, 25, 30, 35 and 40 min after starting measurement. The redox map was obtained by the slope of the enhanced DNP image intensity of each pixel from four pharmacokinetic images using a custom Excel macro program. Phantom redox experiments were conducted to examine the ability of CmP to participate in the mitochondrial redox reaction, and time-dependent changes in image contrast were assessed using liver tissue homogenate and KCN, an inhibitor of mitochondrial electron transfer chain. The DNP images clearly showed the significant enhancement in tissue homogenate solution by CmP. The enhancement of DNP-image was gradually reduced by liver homogenate solution due to reduction of CmP and the reduction rate of CmP was increased by the treatment of radiation. Interestingly, reduction of CmP in both homogenates with or without radiation were completely inhibited by KCN treatment. It is suggested that enhancement of CmP reduction is related in mitochondrial electron transfer system. We demonstrated that the mitochondrial redox metabolic change in the electron transfer chain after radiotherapy could be monitored by DNP-MRI with CmP as a redox probe.

Assessing the influence of isoflurane anesthesia on cardiac metabolism using hyperpolarized [1-13 C]pyruvate.

Isoflurane is a frequently used anesthetic in small-animal dissolution dynamic nuclear polarization-magnetic resonance imaging (DNP-MRI) studies. Although the literature suggests interactions with mitochondrial metabolism, the influence of the compound on cardiac metabolism has not been assessed systematically to date. In the present study, the impact of low versus high isoflurane concentration was examined in a crossover experiment in healthy rats. The results revealed that cardiac metabolism is modulated by isoflurane concentration, showing increased [1-13 C]lactate and reduced [13 C]bicarbonate production during high isoflurane relative to low isoflurane dose [average differences: +16% [1-13 C]lactate/total myocardial carbon, -22% [13 C]bicarbonate/total myocardial carbon; +51% [1-13 C]lactate/[13 C]bicarbonate]. These findings emphasize that reproducible anesthesia is important when studying cardiac metabolism. As the depth of anesthesia is difficult to control in an experimental animal setting, careful study design is required to exclude confounding factors.

In vivo redox metabolic imaging of mitochondria assesses disease progression in non-alcoholic steatohepatitis

Given the rising incidence of non-alcoholic fatty liver disease (NAFLD) in both adults and children, the development of a non-invasive diagnostic method for assessing disease progression to non-alcoholic steatohepatitis (NASH) has become an important research goal. Currently available non-invasive imaging technologies are only able to assess fat accumulation in the liver. Therefore, these methods are not suitable for a precise diagnosis of NASH. The standard diagnostic technique for NASH, liver biopsy, has several drawbacks, including the higher risk of complications that accompanies invasive procedures. Here, we demonstrated that in vivo mitochondrial redox metabolism was dramatically altered at an early stage, before histopathological changes, and NASH could be accurately diagnosed by in vivo dynamic nuclear polarization-magnetic resonance imaging, with carbamoyl-PROXYL as a molecular imaging probe. In addition, this technique was feasible for the diagnosis of NASH compared with histopathological findings from biopsies. Our data reveal a novel method for monitoring the dynamics of redox metabolic changes in NAFLD/NASH.

Sa1592 - The Assessment of Mitochondrial Redox Metabolism by in vivo Dynamic Nuclear Polarization-Magnetic Resonance Imaging for Non-Alcoholic Steatohepatitis

Sa1592 - The Assessment of Mitochondrial Redox Metabolism by in vivo Dynamic Nuclear Polarization-Magnetic Resonance Imaging for Non-Alcoholic Steatohepatitis

Non-invasive evaluation of atopic dermatitis based on redox status using in vivo dynamic nuclear polarization magnetic resonance imaging

Atopic dermatitis (AD) is a chronic inflammatory condition with complex etiology, including genetic, environmental and immunologic factors. Redox imbalance caused by excessive oxidative stress has been shown to mediate disease activity of AD. Currently, an imaging technique that can monitor the redox status of the skin in vivo has not yet been developed. Consequently, we have established such a technique that can detect and visualize the redox status of the skin using in vivo dynamic nuclear polarization magnetic resonance imaging (DNP-MRI). To evaluate this technique, we utilized an AD mouse model that was generated by repeated topical application of mite antigen in NC/Nga mice. We imaged alterations in redox balance of the resulting AD skin lesions of the mice. Using in vivo DNP-MRI and non-toxic nitroxyl radicals to visualize free radicals in vivo, we revealed that AD skin lesions demonstrated more rapid decay rates of image intensity enhancement than normal skin, indicating that our technique can monitor excessive oxidative stress occurring in AD skin lesions. Therefore, this technique has the potential to provide a novel approach for evaluating disease activity of inflammatory skin diseases, including AD, from the view point of altered redox status.

Noninvasive mapping of the redox status of dimethylnitrosamine-induced hepatic fibrosis using in vivo dynamic nuclear polarization-magnetic resonance imaging.

Hepatic fibrosis is a chronic disorder caused by viral infection and/or metabolic, genetic and cholestatic disorders. A noninvasive procedure that enables the detection of liver fibrosis based on redox status would be useful for disease identification and monitoring, and the development of treatments. However, an appropriate technique has not been reported. This study describes a novel method for assessing the redox status of the liver using in vivo dynamic nuclear polarization-magnetic resonance imaging (DNP-MRI) with the nitroxyl radical carbamoyl-PROXYL as a molecular imaging probe, which was tested in dimethylnitrosamine-treated mice as a model of liver fibrosis. Based on the pharmacokinetics of carbamoyl-PROXYL in control livers, reduction rate mapping was performed in fibrotic livers. Reduction rate maps demonstrated a clear difference between the redox status of control and fibrotic livers according to the expression of antioxidants. These findings indicate that in vivo DNP-MRI with a nitroxyl radical probe enables noninvasive detection of changes in liver redox status.

Development of Redox Metabolic Imaging Using Endogenous Molecules.

Redox metabolism plays a central role in maintaining homeostasis in living organisms. The electron transfer system in mitochondria produces ATP via endogenous redox molecules such as flavin mononucleotide (FMN), flavin adenine dinucleotide (FAD), and coenzyme Q10 (CoQ10), which have flavin or quinone moieties. One-electron transfer reactions convert FMN, FAD, and CoQ10 to the free radical intermediates FMNH and FADH, and CoQ10H, respectively. Dynamic nuclear polarization-magnetic resonance imaging (DNP-MRI) allows us to visualize free radicals in vitro and in vivo. We present a spectroscopic imaging technology with DNP-MRI, which enables the imaging of multiple free radical intermediates such as FADH and CoQH. DNP-MRI can also identify various endogenous free radical intermediates derived from redox transformations.

Evaluation of Brain Redox Status and Its Association with Cognitive Dysfunction in Diabetic Animal Models by Redox Molecular Imaging (ReMI).

Oxidative stress contributes to the development of diabetic complications. Increasing epidemiologic evidence suggests that diabetes mellitus is associated with dementia and cognitive decline. However, the underlying mechanisms are not fully understood. We have evaluated brain redox status and its association of cognitive dysfunction in diabetic animal models by dynamic nuclear polarization-magnetic resonance imaging (DNP-MRI) and other oxidative stress markers. In this review, we discuss the role of oxidative stress in the development of diabetes-related dementia and clinical regulation of the redox state in new approaches to augmenting diabetes-related dementia.

Dynamic nuclear polarization-magnetic resonance imaging at low ESR irradiation frequency for ascorbyl free radicals.

Highly water-soluble ubiquinone-0 (CoQ0) reacts with ascorbate monoanion (Asc) to mediate the production of ascorbyl free radicals (AFR). Using aqueous reaction mixture of CoQ0 and Asc, we obtained positively enhanced dynamic nuclear polarization (DNP)-magnetic resonance (MR) images of the AFR at low frequency (ranging from 515 to 530 MHz) of electron spin resonance (ESR) irradiation. The shape of the determined DNP spectrum was similar to ESR absorption spectra with doublet spectral peaks. The relative locational relationship of spectral peaks in the DNP spectra between the AFR (520 and 525 MHz), 14N-labeled carbamoyl-PROXYL (14N-CmP) (526.5 MHz), and Oxo63 (522 MHz) was different from that in the X-band ESR spectra, but were similar to that in the 300-MHz ESR spectra. The ratio of DNP enhancement to radical concentration for the AFR was higher than those for 14N-CmP, Oxo63, and flavin semiquinone radicals. The spectroscopic DNP properties observed for the AFR were essentially the same as those for AFR mediated by pyrroloquinoline quinone. Moreover, we made a success of in vivo DNP-MR imaging of the CoQ0-mediated AFR which was administered by the subcutaneous and oral injections as an imaging probe.

Redox imaging of skeletal muscle using in vivo DNP-MRI and its application to an animal model of local inflammation

Disorders of skeletal muscle are often associated with inflammation and alterations in redox status. A non-invasive technique that could localize and evaluate the severity of skeletal muscle inflammation based on its redox environment would be useful for disease identification and monitoring, and for the development of treatments; however, no such technique currently exists. We describe a method for redox imaging of skeletal muscle using dynamic nuclear polarization magnetic resonance imaging (DNP-MRI), and apply this method to an animal model of local inflammation. Female C57/BL6 mice received injections of 0.5% bupivacaine into their gastrocnemius muscles. Plasma biomarkers, myeloperoxidase activity, and histological sections were assessed at 4 and 24h after bupivacaine injection to measure the inflammatory response. In vivo DNP-MRI was performed with the nitroxyl radicals carbamoyl-PROXYL (cell permeable) and carboxy-PROXYL (cell impermeable) as molecular imaging probes at 4 and 24h after bupivacaine administration. The images obtained after carbamoyl-PROXYL administration were confirmed with the results of L-band EPR spectroscopy. The plasma biomarkers, myeloperoxidase activity, and histological findings indicated that bupivacaine injection caused acute muscle damage and inflammation. DNP-MRI images of mice treated with carbamoyl-PROXYL or carboxy-PROXYL at 4 and 24h after bupivacaine injection showed similar increases in image intensity and decay rate was significantly increased at 24h. In addition, reduction rates in individual mice at 4h and 24h showed faster trends with bupivacaine injection than in their contralateral sides by image-based analysis. These findings indicate that in vivo DNP-MRI with nitroxyl radicals can non-invasively detect changes in the focal redox status of muscle resulting from locally-induced inflammation.

Development of a new redox molecular imaging method

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.

Free Radical Imaging Using In Vivo Dynamic Nuclear Polarization-MRI.

Redox reactions that generate free radical intermediates are essential to metabolic processes, and their intermediates can produce reactive oxygen species, which may promote diseases related to oxidative stress. The development of an in vivo electron spin resonance (ESR) spectrometer and its imaging enables us noninvasive and direct measurement of in vivo free radical reactions in living organisms. The dynamic nuclear polarization magnetic resonance imaging (DNP-MRI), also called PEDRI or OMRI, is also a new imaging method for observing free radical species in vivo. The spatiotemporal resolution of free radical imaging with DNP-MRI is comparable with that in MRI, and each of the radical species can be distinguished in the spectroscopic images by changing the frequency or magnetic field of ESR irradiation. Several kinds of stable nitroxyl radicals were used as spin probes to detect in vivo redox reactions. The signal decay of nitroxyl probes, which is determined with in vivo DNP-MRI, reflects the redox status under oxidative stress, and the signal decay is suppressed by prior administration of antioxidants. In addition, DNP-MRI can also visualize various intermediate free radicals from the intrinsic redox molecules. This noninvasive method, in vivo DNP-MRI, could become a useful tool for investigating the mechanism of oxidative injuries in animal disease models and the in vivo effects of antioxidant drugs.