Fluorodopa is a Promising Fluorine-19 MRI Probe for Evaluating Striatal Dopaminergic Function in a Rat Model of Parkinson's Disease.

Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra projecting to the striatum. It has been estimated that approximately 80% of the striatal dopamine and 50% of nigral dopaminergic neurons are lost before the onset of typical motor symptoms, indicating that early diagnosis of PD using noninvasive imaging is feasible. Fluorine-19 (19 F) magnetic resonance imaging (MRI) represents a highly sensitive, easily available, low-background, and cost-effective approach to evaluate dopaminergic function using non-radioactive fluorine-containing dopaminergic agents. The aim of this study was to find a potent 19 F MRI probe to evaluate dopaminergic presynaptic function in the striatum. To select candidates for 19 F MRI probes, we investigated the following eight non-radioactive fluorine-containing dopaminergic agents: fluorodopa (F-DOPA), F-tyrosine, haloperidol, GBR13069 duhydrochloride, GBR12909 duhydrochloride, 3-bis-(4-fluorophenyl) methoxytropane hydrochloride, flupenthixol, and fenfluramine. In 19 F nuclear magnetic resonance measurements, F-tyrosine and F-DOPA displayed a relatively higher signal-to-noise ratio value in brain homogenates than in others. F-DOPA, but not F-tyrosine, induced the rotational behavior in a 6-hydroxydopamine (6-OHDA)-induced hemiparkinsonian rat model. In addition, a significantly high amount of F-DOPA accumulated in the ipsilateral striatum of hemiparkinsonian rats after the injection. We performed 19 F MRI in PC12 cells and isolated rat brain using a 7T MR scanner. Our findings suggest that F-DOPA is a promising 19 F MRI probe for evaluating dopaminergic presynaptic function in the striatum of hemiparkinsonian rats. © 2016 Wiley Periodicals, Inc.

An MR comparison study of cardiogenic and noncardiogenic pulmonary edema in animal models.

PURPOSE: To apply magnetic resonance (MR) imaging to differential diagnosis of cardiogenic pulmonary edema (CPE) and noncardiogenic pulmonary edema (NCPE). MATERIALS AND METHODS: In the CPE group, MR measurements were performed on 5 rats just before and 3 h after administration of 21 ± 2% body weight of normal saline. In the NCPE group, measurements were similarly performed on 5 animals just before and 48 h after 0.75 mg/body of lipopolysaccharide intratracheal administration. Animals were killed for bronchoalveolar lavage fluid (BALF) or pathological analysis after MR measurements. RESULTS: The relation between signal intensity and the T(2)* value of MR imaging was different in the two groups. The T(2)* in the NCPE group was significantly longer than that in the CPE group at the same signal intensity level. The lactate levels measured by in vivo MR spectroscopy did not show a difference between the CPE and NCPE groups, although the lactate BALF levels of the two groups were significantly different. CONCLUSION: The relation between signal intensity and T(2)* value was useful for the differentiation between these two pulmonary diseases. The measurable lactate level in pulmonary lesions suggests the applicability of MR spectroscopy to pulmonary disease.

Combination of two fat saturation pulses improves detectability of glucose signals in carbon-13 MR spectroscopy.

In order to improve the fat suppression performance of in vivo (13)C-MRS operating at 3.0 Tesla, a phantom model study was conducted using a combination of two fat suppression techniques; a set of pulses for frequency (chemical shift) selective suppression (CHESS), and spatial saturation (SAT). By optimizing the slab thickness for SAT and the irradiation bandwidth for CHESS, the signals of the -(13)CH(3) peak at 49 ppm and the -(13)CH(2)- peak at 26 ppm simulating fat components were suppressed to 5% and 19%, respectively. Combination of these two fat suppression pulses achieved a 53% increase of the height ratio of the glucose C1ß peak compared with the sum of all other peaks, indicating better sensitivity for glucose signal detection. This method will be applicable for in vivo (13)C-MRS by additional adjustment with the in vivo relaxation times of the metabolites.

Simple PEG conjugation of SPIO via an Au-S bond improves its tumor targeting potency as a novel MR tumor imaging agent.

Gold/iron oxide magnetic nanoparticles are hybrid nanoparticles containing a core of magnetic iron oxide and surface colloidal gold, which allows for various biomaterials to be immobilized on the surface of the iron oxide nanoparticles via colloidal gold. Here, we developed a novel magnetic resonance (MR) imaging agent to broaden the MR tumor-imaging spectrum of superparamagnetic iron oxide nanoparticles (SPIO), e.g., Feridex(), a clinical MR imaging agent for diagnosing liver cancer. Au/Feridex was synthesized by electron beam irradiation, and thiol-modified poly(ethylene glycol) (PEG-SH) was easily conjugated to its surface via an Au-S bond without the need for any chemical reactions. PEG conjugation of Au/Feridex enhanced its accumulation in Meth-A tumor tissue and decreased its accumulation in normal liver tissue. In addition, MRI using PEG-Au/Feridex, in contrast to MRI using unmodified Au/Feridex and Feridex, detected B16BL6 and Meth-A tumor tissues in vivo. This finding indicates that PEG-Au/Feridex is useful for diagnosing various types of tumors. In addition, because the synthesis of PEG-Au/Feridex is simple and high yields are easily produced, PEG-modified SPIO for tumor diagnosis can be prepared on an industrial scale with low cost.

Clinical features of myocardial triglyceride in different types of cardiomyopathy assessed by proton magnetic resonance spectroscopy: comparison with myocardial creatine.

BACKGROUND: Myocardial lipid overstorage may produce cardiomyopathy, leading to dysfunction, but advanced heart failure may cause lipolysis via sympathetic nerve activation. In the failing heart, the creatine kinase system may also be impaired. The aims of this study were to assess myocardial triglyceride (TG) and creatine (CR) in different types of cardiomyopathy and to investigate whether they are related to the severity of cardiac dysfunction. METHODS AND RESULTS: In patients with hypertrophic cardiomyopathy (HCM, n = 8), dilated cardiomyopathy (DCM, n = 12) or ischemic cardiomyopathy (ICM, n = 10), and normal subjects (NML, n = 22), myocardial TG and CR were evaluated using proton magnetic resonance spectroscopy. To assess cardiac sympathetic nerve activity, myocardial MIBG (a radioactive guanethidine analog) uptake was measured in DCM. Myocardial TG was significantly lower in hypertrophic cardiomyopathy (HCM) (1.92 ± 0.99 µmol/g), but higher in ICM (7.59 ± 4.36 µmol/g) than in NML hearts (4.05 ± 1.94 µmol/g). There was no significant difference in TG between DCM (4.84 ± 6.45 µmol/g) and NML. Myocardial CR in HCM (20.4 ± 8.4 µmol/g), DCM (14.8 ± 4.8 µmol/g), and ICM (19.4 ± 6.3 µmol/g) was significantly lower than that in NML hearts (27.1 ± 4.3 µmol/g). Overall, myocardial CR correlated positively with the severity of heart failure estimated by ejection fraction or myocardial BMIPP (a radioactive fatty acid analog) uptake, but TG did not. In DCM, myocardial TG correlated with body mass index, but not with MIBG uptake. CONCLUSIONS: Myocardial TG may be related to the specific cause of disease rather than the severity of cardiac dysfunction. In contrast, myocardial CR reflects the severity of heart failure despite different pathoetiologic mechanisms of dysfunction. In DCM, myocardial TG may be affected by an overweight state rather than cardiac sympathetic nerve dysfunction. Thus, myocardial CR has a closer relationship to heart failure severity than does myocardial TG.

Preparation for highly sensitive MRI contrast agents using core/shell type nanoparticles consisting of multiple SPIO cores with thin silica coating.

We describe here the facile and robust preparation methods for the multiple-SPIO-containing silica-coated core/shell type nanoparticles which can serve as a highly sensitive MRI contrast agent. The imidazolium-tethered core/shell type particles were synthesized, and the centrifugal selection for the multiple-SPIO-containing particles and the etching process to fabricate thin silica layers were carried out to improve the proton relaxivity of water tissue. We found that the synthetic particles can provide approximately 7-fold clearer contrasts than that of the particles before treatments. In addition, the particles can show good dispersibility at least for 1 week in aqueous media.

Correlation between musculoskeletal structure of the hand and primate locomotion: Morphometric and mechanical analysis in prehension using the cross- and triple-ratios.

Biometric ratios of the relative length of the rays in the hand have been analyzed between primate species in the light of their hand function or phylogeny. However, how relative lengths among phalanges are mechanically linked to the grasping function of primates with different locomotor behaviors remains unclear. To clarify this, we calculated cross and triple-ratios, which are related to the torque distribution, and the torque generation mode at different joint angles using the lengths of the phalanges and metacarpal bones in 52 primates belonging to 25 species. The torque exerted on the finger joint and traction force of the flexor tendons necessary for a cylindrical grip and a suspensory hand posture were calculated using the moment arm of flexor tendons measured on magnetic resonance images, and were compared among Hylobates spp., Ateles sp., and Papio hamadryas. Finally, the torques calculated from the model were validated by a mechanical study detecting the force exerted on the phalanx by pulling the digital flexor muscles during suspension in these three species. Canonical discriminant analysis of cross and triple-ratios classified primates almost in accordance with their current classification based on locomotor behavior. The traction force was markedly reduced with flexion of the MCP joint parallel to the torque in brachiating primates; this was notably lower in the terrestrial quadrupedal primates than in the arboreal primates at mild flexion. Our mechanical study supported these features in the torque and traction force generation efficiencies. Our results suggest that suspensory or terrestrial quadrupedal primates have hand structures that can exert more torque at a suspensory posture, or palmigrade and digitigrade locomotion, respectively. Furthermore, our study suggests availability of the cross and triple-ratios as one of the indicators to estimate the hand function from the skeletal structure.

DJ-1 protects against neurodegeneration caused by focal cerebral ischemia and reperfusion in rats.

Reactive oxygen species (ROS) is massively produced in the brain after cerebral ischemia and reperfusion. It reacts strongly with cellular components, which has detrimental effects and leads to neuronal cell death. DJ-1, which was found to be the causative gene of familial Parkinson's disease PARK7, is a multifunction protein, which plays a key role in transcriptional regulation, and a molecular chaperone. In this study, we investigated the neuroprotective effect of DJ-1 against neurodegeneration caused by ischemia/reperfusion injury. Cerebral ischemia was induced in rats by 120 mins of middle cerebral artery occlusion (MCAO) using an intraluminal introduction method. The intrastriatal injection of recombinant glutathione S-transferase-tagged human DJ-1 (GST-DJ-1) markedly reduced infarct size in 2,3,5-triphenyltetrazolium chloride staining at 3 days after MCAO. In addition, we performed a noninvasive evaluation of ischemic size using magnetic resonance imaging and found a significant reduction of infarct size with the administration of GST-DJ-1. In GST-DJ-1-treated rats, behavioral dysfunction and nitrotyrosine formation were significantly inhibited. Furthermore, GST-DJ-1 markedly inhibited H(2)O(2)-mediated ROS production in SH-SY5Y cells. These results indicate that GST-DJ-1 exerts a neuroprotective effect by reducing ROS-mediated neuronal injury, suggesting that DJ-1 may be a useful therapeutic target for ischemic neurodegeneration.

Assembly system of direct modified superparamagnetic iron oxide nanoparticles for target-specific MRI contrast agents.

We report the direct modification of SPIOs with a biomolecule and the target-specific assembly system of these modified SPIOs for using MRI contrast agents. The transverse relaxation rate of the aqueous solutions containing the modified SPIOs was altered by the dispersion state.

Correlation between high field MR images and histopathological findings of rat transplanted cancer immediately after partial microwave coagulation.

PURPOSE: To investigate the immediate effects of microwave coagulation on rat tumors in various magnetic resonance (MR) images at high magnetic field strength using histopathological examinations as reference. MATERIALS AND METHODS: Tumors implanted in rat femurs were partially thermocoagulated by microwave. Immediately after, T1- and T2-weighted images, diffusion-weighted images (DWIs), and contrast-enhanced T1 weighted images (CE-T1WIs) were acquired with a 7-tesla MR scanner. After measurements, tumors were examined histopathologically with hematoxylin-eosin (HE) staining and histochemically for acid phosphatase activity. RESULTS: Without contrast, boundaries of coagulated areas were unclear on MR images, including apparent diffusion coefficient (ADC) maps. CE-T1WIs clearly showed immediate contrast enhancement of untreated areas of tumor, and the area of enhancement gradually enlarged in 5 min. Quantitative analyses were conducted by classifying tumor areas by contrast enhancement results. Signal intensities of the areas in the MR images showed no significant differences, but at the periphery, ADC values were significantly higher in areas with delayed enhancement than those with immediate enhancement. Compared with histopathological findings, with microwave thermocoagulation, increased ADC value seemed to derive from collection of extracellular fluid in the outer zone, where acid phosphatase activity was attenuated. CONCLUSION: ADC values in the areas with delayed enhancement of CE-T1WIs were higher than those in non-affected areas, but MR images could not show areas of coagulation within tumors. Clear detection of the boundaries of coagulated areas required contrast enhancement, even at magnetic field strength of 7T.

Microglial transplantation increases amyloid-beta clearance in Alzheimer model rats.

Immunization with amyloid-beta (Abeta) peptides, a therapeutic approach in Alzheimer's disease (AD), reduces brain Abeta, and microglial Abeta phagocytosis has been proposed as an Abeta-lowering mechanism. We transplanted rat microglia into the rat lateral ventricle just after intra-hippocampal Abeta injection, and then investigated the contribution of exogenous microglia to Abeta clearance. Migration of exogenous microglia from the lateral ventricle to Abeta plaque was detected by magnetic resonance imaging and histochemical analysis, and the clearance of Abeta was increased by transplantation. These results suggest the possible usefulness of exogenous microglia to the therapeutic approach in AD.

The MR tracking of transplanted ATDC5 cells using fluorinated poly-L-lysine-CF3.

Magnetic resonance (MR) imaging using super-paramagnetic iron oxides (SPIOs) is a powerful tool to monitor transplanted cells in living animals. However, since SPIOs are negative contrast agents it is difficult to track transplanted cells in bone and cartilage that originally display low signals. In this study, we examined the feasibility of tracking with fluorescein isothiocyanate (FITC)-labeled poly-L-lysine-CF(3) (PLK-CF(3)) using mouse ATDC5 cells, a stem cell line of bone and cartilage cells. FITC-labeled PLK-CF(3) was easily internalized by ATDC5 cells by adding it into culture medium. No acute or long-term toxicities were seen at less than 160 microg/ml. Labeled cells transplanted into the cranial bone of mice were detected for at least 7 days by MR images. FITC-labeled PLK-CF(3) is a useful positive contrast agent for MR tracking in bone and cartilage.

Fluorine-19 fast recovery fast spin echo imaging for mapping 5-fluorouracil.

We investigated the effects of fast recovery (FR) to increase the sensitivity of fluorine-19 ((19)F) fast spin echo (FSE) in mapping 5-fluorouracil (5-FU) and its metabolites. We added an additional 90 degrees pulse (which flips back longitudinal magnetization at the end of the sequence) to the chemical shift selective (19)F FSE pulse sequence. In 5-FU solution, FR remarkably improved the signal-to-noise (S/N) ratio of (19)F 5-FU images, having higher effects with shorter repetition time and smaller echo train numbers. In animal studies, FR produced a conspicuous increase in (19)F signals in the urinary bladder. FR effects for (19)F signals in the liver were smaller than those in other organs but still substantial. Utilization of FR in (19)F FSE images promises more sensitive observation of (19)F metabolite maps of 5-FU and other (19)F-containing compounds that have relatively long relaxation times.

Low blood flow estimates in lower-leg arteries predict cardiovascular events in Japanese patients with type 2 diabetes with normal ankle-brachial indexes.

OBJECTIVE: To examine the association of baseline measures in lower-leg arteries and conventional cardiovascular risk factors with the incidence of cardiovascular disease (CVD) events in type 2 diabetic patients with normal ankle-brachial indexes (ABIs) (>0.9). RESEARCH DESIGN AND METHODS: We studied 129 type 2 diabetic patients and 35 age-matched nondiabetic subjects with no apparent CVD consecutively admitted to our hospital. At baseline, total flow volume and resistive index, as an index of vascular resistance, at the popliteal artery was evaluated using gated two-dimensional cine-mode phase-contrast magnetic resonance imaging. Patients were followed 4.8 +/- 1.5 years (range 3.0-8.2) or until their first event of CVD. RESULTS: On follow-up, 16 patients developed primary CVD events. Patients with CVD had lower blood flow (P < 0.01) and higher vascular resistance (P < 0.05) than patients without CVD. When the patients were grouped into tertiles according to their levels of total flow volume (129.6-85.5, 85.3-63.3, and 62.7-23.8 ml/min), Kaplan-Meier analysis showed a higher probability of developing CVD events in patients in the lowest than in patients in the highest (P = 0.0199, log-rank test) tertile. Multivariate Cox proportional hazards analysis revealed that the lowest tertile for flow volume (hazard ratio [HR] 8.60, 95% CI 1.61-45.97, P = 0.012), hypertension (3.99, 1.12-14.25, P = 0.033), and smoking status (12.01, 1.21-119.28, P = 0.034) were significant independent predictors of CVD events. CONCLUSIONS: We have demonstrated that low blood flow estimates in lower-leg arteries may be predictive for CVD events among Japanese patients with type 2 diabetes even though they have a normal ABI.

Transplantation of iPS-Derived Tumor Cells with a Homozygous MHC Haplotype Induces GRP94 Antibody Production in MHC-Matched Macaques.

Immune surveillance is a critical component of the antitumor response in vivo, yet the specific components of the immune system involved in this regulatory response remain unclear. In this study, we demonstrate that autoantibodies can mitigate tumor growth in vitro and in vivo We generated two cancer cell lines, embryonal carcinoma and glioblastoma cell lines, from monkey-induced pluripotent stem cells (iPSC) carrying a homozygous haplotype of major histocompatibility complex (MHC, Mafa in Macaca fascicularis). To establish a monkey cancer model, we transplanted these cells into monkeys carrying the matched Mafa haplotype in one of the chromosomes. Neither Mafa-homozygous cancer cell line grew in monkeys carrying the matched Mafa haplotype heterozygously. We detected in the plasma of these monkeys an IgG autoantibody against GRP94, a heat shock protein. Injection of the plasma prevented growth of the tumor cells in immunodeficient mice, whereas plasma IgG depleted of GRP94 IgG exhibited reduced killing activity against cancer cells in vitro These results indicate that humoral immunity, including autoantibodies against GRP94, plays a role in cancer immune surveillance. Cancer Res; 77(21); 6001-10. ©2017 AACR.

MR tracking of transplanted glial cells using poly-L-lysine-CF3.

Magnetic resonance (MR) imaging using super-paramagnetic iron oxides (SPIOs) is a powerful tool to monitor transplanted cells in living animals. Since, however, SPIOs are negative contrast agents, positive agents have been explored. In this study, we examined the feasibility of FITC-labeled poly-L-lysine-CF3 (PLK-CF3) using glial cells. FITC-labeled PLK-CF3 was easily internalized by neuroblastoma cells and glia as adding it into culture medium. No toxicity was seen at the concentration of less than 80 microg/ml. MR images positively detected labeled cells transplanted in the brain of living mouse. The results indicate that FITC-labeled PLK-CF3 is a useful positive contrast agent for MR tracking.

Magnetic resonance imaging using hemagglutinating virus of Japan-envelope vector successfully detects localization of intra-cardially administered microglia in normal mouse brain.

Although the therapeutic use of microglia has received some attention for the treatment of brain diseases, few non-invasive techniques exist for monitoring the cells after administration. Here, we present a technique using magnetic resonance imaging (MRI) to track microglia injected intra-cardially. We labeled microglia expressing enhanced green fluorescent protein with superparamagnetic iron oxide (Resovist) using the hemagglutinating virus of Japan-envelope vector. We injected labeled microglia into the left ventricle of the heart of mice. After monitoring exogenously administered microglia in the mouse brain in vivo using T(2)*-weighted MRI at a magnetic field of 7T, we compared the MR images with histochemical localization of exogenous microglia in vitro. MRI revealed clear signal changes attributable to Resovist-containing microglia in the mouse brain. Histochemistry demonstrated the presence of exogenous microglia in the brain at the same locations shown by MRI. This study demonstrates the usefulness of MRI for non-invasive monitoring of exogenous microglia, and suggests a promising future for microglia/macrophages as therapeutic tools for brain disease.

N-Acetylaspartate concentrations in the thalami of neuropathic pain patients and healthy comparison subjects measured with (1)H-MRS.

We investigated the absolute concentration of N-acetylaspartate (NAA) in the thalami of neuropathic pain patients and healthy comparison subjects by single-voxel proton magnetic resonance spectroscopy ((1)H-MRS). (1)H-MRS was performed with a 1.5-T MR system on a voxel in the thalamus bilaterally in 9 neuropathic pain patients and 14 healthy control subjects. We measured the absolute concentration of NAA using a linear combination model. The NAA concentration in the thalamus decreased significantly on the contralateral side in seven patients and on the ipsilateral side in two patients, as compared with the mean NAA concentration of the healthy control subjects. The NAA concentrations in two patients who did not respond to standard pain treatments were extremely decreased. Our results using (1)H-MRS suggest that neuropathic pain seems to be associated with an abnormal balance of the neural activity in the thalamus. The NAA concentration of the thalamus may be related to the efficacy of therapy. (1)H-MRS may serve as a useful noninvasive tool for evaluating thalamic neural activity in neuropathic pain patients.

[Plasma platinum concentration and anti-tumor effects after intra-arterial infusion of lipiodol-CDDP suspension evaluation with VX 2 rabbit liver cancer model and 7.0 Tesla MRI system].

To evaluate the usefulness of combination chemotherapy with cisplatin powder, which was newly designed for intra-arterial infusion (IA CALL) and lipiodol, for the VX 2 liver cancer model of the rabbit, sequential change of the plasma platinum concentration within the first 24 hours, as well as the tissue platinum concentration at 24 hours was measured after intra-arterial infusion of IA CALL. The infused materials were either IA CALL alone (C) or the combination of lipiodol+IA CALL (CL). In addition,the reduction rate of the VX 2 tumor was calculated among four therapeutic groups (C, CL, lipiodol alone (L), and saline alone (S)) after one week of intra-arterial infusion on the basis of 7.0 Tesla MR images. Total plasma platinum concentrations within the first 24 hours were kept low in group CL. No increase in the tissue platinum concentration in group CL was observed. On the other hand, the tumor reduction rate tended to be higher in group CL (group CL>group L=group C>group S). These results suggested that the intra-arterial infusion of IA CALL with lipiodol is more effective than that of IA CALL alone.

Amyloid imaging using fluorine-19 magnetic resonance imaging ((19)F-MRI).

The formation of senile plaques followed by the deposition of amyloid-ß is the earliest pathological change in Alzheimer's disease. Thus, the detection of senile plaques remains the most important early diagnostic indicator of Alzheimer's disease. Amyloid imaging is a noninvasive technique for visualizing senile plaques in the brains of Alzheimer's patients using positron emission tomography (PET) or magnetic resonance imaging (MRI). Because fluorine-19 ((19)F) displays an intense nuclear magnetic resonance signal and is almost non-existent in the body, targets are detected with a higher signal-to-noise ratio using appropriate fluorinated contrast agents. The recent introduction of high-field MRI allows us to detect amyloid depositions in the brain of living mouse using (19)F-MRI. So far, at least three probes have been reported to detect amyloid deposition in the brain of transgenic mouse models of Alzheimer's disease; (E,E)-1-fluoro-2,5-bis-(3-hydroxycarbonyl-4-hydroxy)styrylbenzene (FSB), 1,7-bis(4'-hydroxy-3'-trifluoromethoxyphenyl)-4-methoxycarbonylethyl-1,6-heptadiene3,5-dione (FMeC1, Shiga-Y5) and 6-(3',6',9',15',18',21'-heptaoxa-23',23',23'-trifluorotricosanyloxy)-2-(4'-dimethylaminostyryl)benzoxazole (XP7, Shiga-X22). This review presents the recent advances in amyloid imaging using (19)F-MRI, including our own studies.