Visualization of in vivo electroporation-mediated transgene expression in experimental tumors by optical and magnetic resonance imaging.

In vivo electroporation (EP) is an efficient method for effective gene transfer and is highly expected for application in anticancer gene therapy. Non-invasive monitoring of gene transfer/expression is critical for optimal gene therapy. Here we report in vivo optical and high-field magnetic resonance imaging (MRI) of EP-mediated transgene expression in a tumor model. Initially, we observed spatio-temporal change in in vivo EP-mediated transgene expression by optical imaging using red fluorescence protein (RFP) as a reporter gene. Next, we constructed a dual-reporter plasmid carrying a gene-encoding MRI reporter ferritin heavy chain and RFP gene to visualize the intratumoral transgene expression by dual modality. Cells transfected with this plasmid showed lower signal intensity on in vitro T(2)-weighted cellular MRI and quantitatively increased the transverse relaxation rate (1/T(2)) compared with control cells. After conducting in vivo EP in an experimental tumor, the plasmid-injected region showed both fluorescent emissions in optical imaging and detectably lowered signal on T(2)-weighted MRI. The correlative immunohistological findings confirmed that both the reporter transgenes were co-expressed in this region. Thus, our strategy provides a platform for evaluating EP-mediated cancer gene therapy easily and safely without administering contrast agent or substrate.

Degeneration of patellar cartilage in patients with recurrent patellar dislocation following conservative treatment: evaluation with delayed gadolinium-enhanced magnetic resonance imaging of cartilage.

OBJECTIVE: To examine the characteristics of cartilage degeneration in patients with recurrent patellar dislocation (RPD) following conservative treatment using delayed gadolinium-enhanced magnetic resonance imaging (MRI) of cartilage (dGEMRIC). DESIGN: This study evaluated three groups of knees: group I, 35 knees from both knees of patients with bilateral RPD and dislocated side knees of patients with unilateral RPD; group II, 15 non-dislocated side knees of patients with unilateral RPD; and group III, 20 knees from both knees of healthy volunteers. Differences in post-contrast T1 [T1(Gd)] of cartilage at both medial and lateral facets between groups I, II and III were analyzed. For group I, possible relationships were evaluated between T1(Gd) of cartilage and patient age, length of time between the initial dislocation and MRI and the total number of dislocations between the initial dislocation and MRI for both medial and lateral facets. RESULTS: The mean T1(Gd) of cartilage at medial facets for groups I, II and III were 411+/-46ms, 465+/-38ms and 490+/-29ms, respectively; there were significant differences between these means (P<0.05). The mean T1(Gd) of cartilage at lateral facets for groups I, II and III were 426+/-53ms, 466+/-45ms and 510+/-36ms, respectively; there were also significant differences between these means (P<0.05). Significant correlations were observed between T1(Gd) of cartilage for both medial and lateral facets and length of time between the initial dislocation and MRI (P<0.05). No other correlations were significant. CONCLUSION: dGEMRIC may be a useful method to monitor glycosaminoglycan concentration in patients with RPD following conservative treatment.

Distinctly abnormal brain metabolism in late-onset ornithine transcarbamylase deficiency.

OBJECTIVE: To assess alterations in brain metabolites in patients with late-onset ornithine transcarbamylase deficiency (OTCD). METHODS: Six unrelated, asymptomatic Japanese late-onset OTCD patients were analyzed by proton MRS ((1)HMRS) using a point-resolved spectroscopy technique (repetition and echo times, 5000 and 30 ms). Localized spectra for the centrum semiovale were acquired and absolute metabolite concentrations were calculated using an LCModel. RESULTS: Compared with age-matched controls, N-acetylaspartate and creatine concentrations were normal in all patients. The glutamine (Gln) plus glutamate concentration was increased in four patients, which progressed in proportion to the clinical stage. myo-inositol (mI) could not be detected in five symptomatic patients. A decreased choline (Cho) concentration was detected in two clinically severe patients. (1)HMRS after liver transplantation in one patient revealed the normalization of all metabolites. CONCLUSION: These findings suggest progression of neurochemical events in OTCD, i.e., mI depletion and Gln accumulation followed by Cho depletion, which is reverse of that in hepatic encephalopathy, i.e., Cho depletion followed by mI depletion and Gln accumulation.

Brain N-acetylaspartate is elevated in Pelizaeus-Merzbacher disease with PLP1 duplication.

OBJECTIVE: To assess alterations in brain metabolites of patients with Pelizaeus-Merzbacher disease (PMD) with the proteolipid protein gene 1 (PLP1) duplications using quantitative proton MRS. METHODS: Five unrelated male Japanese patients with PMD with PLP1 duplications were analyzed using automated proton brain examination with the point resolved spectroscopy technique (repetition and echo time of 5,000 and 30 msec). Localized spectra in the posterior portion of the centrum semiovale were acquired, and absolute metabolite concentrations were calculated using the LCModel. RESULTS: Absolute concentrations of N-acetylaspartate (NAA), creatine (Cr), and myoinositol (MI) were increased by 16% (p < 0.01), 43% (p < 0.001), and 31% (p < 0.01) in patients with PMD as compared with age-matched controls. There was no statistical difference in choline concentration. CONCLUSION: The increased concentration of NAA, which could not be detected by previous relative quantitation methods, suggests two possibilities: axonal involvement secondary to dysmyelination, or increased cell population of oligodendrocyte progenitors. Elevated Cr and MI concentrations may reflect the reactive astrocytic gliosis. Our study thus emphasizes the importance of absolute quantitation of metabolites to investigate the disease mechanism of the dysmyelinating disorders of the CNS.

Long-term assessment of posttransplant renal prognosis with 31 P magnetic resonance spectroscopy.

BACKGROUND: 31P-magnetic resonance spectroscopy (MRS) has been widely used to study pretransplantation renal viability, and although some had discussed posttransplant renal viability, no one has examined long-term posttransplant renal prognosis. We discuss the use of 31P-MRS to assess the long-term prognosis from the time when MRS was performed. METHODS: We studied 20 patients with renal allografts. 1.5 Tesla clinical magnetic resonance imaging (MRI) and 15 cm surface coil was used for 31P-MRS. Localized 31P-MRS was done using image selected in vivo spectroscopy (ISIS) method. Individual peaks were fitted by Lorenzian line-shapes with a least square method and peak area ratios were calculated. RESULTS: A beta-adenosine triphosphate/inorganic phosphate (beta-ATP/Pi) ratio >1.2 had sensitivity of 92.8%, specificity of 100%, and accuracy of 95% for predicting 3-year renal survival; a beta-ATP/Pi ratio >1.2 had sensitivity of 90.9%, specificity of 66.7%, and accuracy of 76.9% for predicting 5-year renal survival. We compared 31P-MRS spectra data between the survived group and failed group. The survived group had significantly higher beta-ATP/Pi, alpha-ATP/Pi, and phosphodiester (PDE)/Pi ratios than the failed group. CONCLUSIONS: We discussed the beta-ATP/Pi value as a parameter for predicting long-term survival of a transplanted kidney from the time when MRS was performed. A value above 1.2 suggests a high probability of 3-year renal survival, whereas a value over 2.5 indicates that the transplanted kidney could survive over 5 years. 31P-MRS may be useful for predicting long-term survival of transplanted kidneys, but additional studies are needed.

Visualization of specific binding sites of benzodiazepine in human brain.

Using 11C-labeled Ro15-1788 and positron emission tomography, studies of benzodiazepine binding sites in the human brain were performed on four normal volunteers. Rapid and high accumulation of 11C activity was observed in the brain after i.v. injection of [11C]Ro15-1788, the maximum of which was within 12 min. Initial distribution of 11C activity in the brain was similar to the distribution of the normal cerebral blood flow. Ten minutes after injection, however, a high uptake of 11C activity was observed in the cerebral cortex and moderate uptake was seen in the cerebellar cortex, the basal ganglia, and the thalamus. The accumulation of 11C activity was low in the brain stem. This distribution of 11C activity was approximately parallel to the known distribution of benzodiazepine receptors. Saturation experiments were performed on four volunteers with oral administration of 0.3-1.8 mg/kg of cold Ro15-1788 prior to injection. Initial distribution of 11C activity following injection peaked within 2 min and then the accumulation of 11C activity decreased rapidly and remarkably throughout the brain. The results indicated that [11C] Ro15-1788 associates and dissociates to specific and nonspecific binding sites rapidly and has a high ratio of specific receptor binding to nonspecific binding in vivo. Carbon-11 Ro15-1788 is a suitable radioligand for the study of benzodiazepine receptors in vivo in humans.

Kinetics of [11C]N,N-dimethylphenylethylamine in mice and humans: potential for measurement of brain MAO-B activity.

Carbon-11-labeled N,N-dimethylphenylethylamine ([11C]DMPEA) was synthesized by the reaction of N-methylphenylethylamine with [11C]methyl iodide. This newly synthesized radiotracer was developed for the purpose of in vivo measurement of monoamine oxidase-B activity in the brain using a metabolic trapping method. Initially, biodistribution was investigated in mice. The rapid and high uptake of 11C radioactivity in the brain was observed following intravenous injection of [11C]DMPEA, the peak of which was reached at 1 min, followed by a decrease at 1-5 min and slowly thereafter. The kinetics of [11C]DMPEA in the human brain were determined using positron emission tomography (PET) and showed that 11C radioactivity increased gradually over 60 min following initial rapid uptake of 11C radioactivity, with basal ganglia and thalamus showing high accumulation.

Proton magnetic resonance imaging of flow motion of heavy water injected into a hollow fiber dialyzer filled with saline.

Observations using MRI were performed for the motion of heavy water injected into a hollow fiber dialyzer. A cylindrical dialyzer houses a bundle of 10,000 hollow fibers. Because blood components permeate through the hollow fiber membrane from the interior to the exterior of the hollow fiber, which is the dialysate flow path, uniformity of dialysate flow is required. The dialyzer was initially filled with saline and heavy water was injected into the inlet port of the dialysate flow path. MRI tuned for protons could distinguish the injected heavy water from the already present saline. Due to the specific gravity difference, MRI could observe the sedimentation of the injected heavy water flowing beneath the already present saline. The uniformity of the dialysate flow was supported by the finding that the injected heavy water brought about uniform sedimentation and distributed the already present saline uniformly throughout the entire volume of the dialyzer.

Detection of small degree of nonuniformity in dialysate flow in hollow-fiber dialyzer using proton magnetic resonance imaging.

A small degree of nonuniformity in dialysate flow in a hollow-fiber dialyzer was detected using proton magnetic resonance imaging (MRI). Since paramagnetic ions reduce the spin-lattice relaxation time of protons around them, MRI can detect Gd in water. An aqueous solution of a chelate compound of Gd was impulsively injected into the dialysate flow path at a flow rate of 500 cm(3) /m, which is that utilized in actual dialysis. Despite the apparent elimination of Gd from the dialysate flow path by the newly injected dialysate fluid after the injection of Gd was terminated, MRI revealed that Gd remained in the interior of the hollow fiber. The observed structure pattern of the Gd concentration profile revealed that the dialysate flow had a small degree of nonuniformity despite the currently established design to restrict channeling in dialysate flow. Local nonuniformity of the hollow-fiber density and vortex generation in the dialysate flow were considered to cause the nonuniformity in the dialysate flow.

Study of biodistribution of enflurane in rats with in vivo 19F MRI.

In vivo 19F magnetic resonance imaging (MRI) of anesthetized rats enabled us to visualize the biodistribution of fluorinated anesthetics and to document the changes in MR signals in the body during the induction and the elimination phase of anesthesia. The authors examined in vivo 19F MRI in rats anesthetized with concentrations of 1.75-2.0% enflurane and demonstrated its in vivo distribution with concomitant 1H and 13C MRI to verify the anatomical correlation. Distinct 19F MR signals were acquired predominantly from the systemic adipose tissue and the liver. Additionally, the temporal changes in the tissue during and after anesthesia were characterized with in vivo 19F MRI in 6.4 min of the acquisition time. The 19F MR signals increased with time after anesthesia; however, the signals from the adipose tissue were apparently stronger than those from the liver. Following the discontinuation of inhalation, the MR signals in the liver decreased far more rapidly than those from the adipose tissue. When the animal woke up and began to move, the MR signals were still visible in the adipose tissue. These results confirmed the fact that enflurane dissolves preferentially in the adipose tissue and remains when the anesthetic effect disappears. Additionally, 19F MR signals of the liver during the elimination phase might reflect the concentration of enflurane in the blood.

Noninvasive analysis of water movement in rat testis using deuterium magnetic resonance imaging.

To measure water movement in the testis without the effects from the blood-testis barrier, we performed in vivo deuterium magnetic resonance imaging (2H MRI) of rats administered with deuterated saline. Alcohol was injected into one testis of each animal and the other was administered with normal saline as a control. Dynamic 2H MRI was obtained at 2 T by FLASH pulse sequence (TR, 300 ms; TE, 10 ms; alpha = 90 degrees) using a surface coil (3 cm in diameter). The variation in 2H signal intensity between the two testes as a function of time after deuterated saline injection was examined every 1.1 min up to 20 min. The signal intensity in the testis receiving the alcohol treatment was lower than that in the normal control. Thus, deuterium MRI can be used to analyze functional disorders of the testis.

Three-vessel study of cerebral blood flow using phase-contrast magnetic resonance imaging: effect of physical characteristics.

The development of phase-contrast magnetic resonance imaging (P-C MRI) provides a noninvasive method for measurement of volumetric blood flow (VFR). We performed P-C MRI to study the effects of physical characteristics on cerebral blood flow. VFR of the left and right internal carotid arteries and basilar artery were measured using P-C MRI and total cerebral blood flow (tCBF) was calculated by summing up the VFR values in the three vessels. Moreover, we investigated the changes in these blood flows as influenced by age, head size, height, weight, body surface area, and handedness. The blood flows were 142 +/- 58 ml/min (mean +/- standard deviation) in the basilar artery; and 229 +/- 86 ml/min in the left, and 223 +/- 58 ml/min in the right internal carotid artery; and tCBF was 617 +/- 128 ml/min. Significant increases were observed in head size-related change of VFR in the basilar artery (p = .028) and height-related change of tCBF (p = .045). The other characteristics did not significantly influence any VFR. The results suggest that head size and height may reflect CBF, and that these effects should be considered when changes of CBF are diagnosed. Phase-contrast MRI is useful for a noninvasive and rapid analysis of cerebral VFR and has potential for clinical use.

Acute pan-costochondritis demonstrated by gallium scintigraphy.

Costochondral inflammation is a rare clinical finding. A case is presented, in which acute chostochondritis is demonstrated by gallium scinitigraphy.

A preliminary study for clinical pharmacokinetics of oral fluorine anticancer medicines using the commercial MRI system 19F-MRS.

These preliminary studies of dynamic natural abundance 19F-magnetic resonance spectroscopy (19F-MRS) on 5-FU based medicines were performed in the human liver using commercial 1.5 T MRI equipment. A single tuned, custom-made circular shape surface coil with a diameter of 15 cm operating at 60 MHz was used for the 19F-MRS study. Localized proton shimming with a whole body coil was performed with adequate volume to include the observing area of the surface coil, and the line width of the water signal was less than 40 Hz. Very different spectroscopic appearance patterns of 5-FU were observed. We examined whether the pharmacokinetics in the liver of orally administered 1-hexylcarbamoyl-5-fluorouracil (HCFU) differ from those of orally administered 5'-deoxy-5-fluorouridine (5'-DFUR). This preliminary study suggested the 19F-MRS technique could be a useful method of evaluating in vivo the metabolism of 5-FU based medicines.

Seizure frequency and bilateral temporal abnormalities: a proton magnetic resonance spectroscopy of temporal lobe epilepsy.

Proton magnetic resonance spectroscopy ((1)H-MRS) was performed in seven healthy volunteers and 17 patients with temporal lobe epilepsy (TLE) to clarify the correlation of the severity of epilepsy with bilateral temporal changes in N-acetylaspartate (NAA), choline-containing compounds (Cho) and creatine + phosphocreatine (Cr). Despite unilateral EEG focus, bilateral temporal reduction in NAA /(Cho + Cr) was revealed in patients with intractable seizures. The potential for seizure generation correlated with the NAA /(Cho + Cr) reduction not only on the ipsilateral side but also on the contralateral side. Proton MRS proved to be a useful measurement for obtaining important information about the neuronal changes as well as the lateralization of the epileptogenic focus in TLE patients.

The measurement of blood flow parameters with deuterium stable isotope MR imaging.

METHODS: Because there are no radioactive hydrogen isotopes which can be used for clinical examinations, deuterium as a non-radioactive, freely diffusible tracer has some advantages compared with the radioactive tracers in the measurement of blood flow parameters. A non-invasive technique to estimate the mean tissue blood flow parameter in vivo was developed by using deuterium nuclear magnetic resonance (NMR) imaging in rat. We obtained the NMR signal changes from deuterium NMR images in nine male Wister rats after intravenous injection of D2O and applied exponential curve fitting analyses to calculate blood flow parameters of the brain, heart and skeletal muscle. RESULTS: While fitting the reducing of the monoexponential function yielded a blood flow parameter of 27.9 +/- 1.6 ml/min/100 g tissue weight for the brain and 46.7 +/- 3.7 ml/min/100 g tissue weight for the heart, fitting the early reducing of the signal intensity of the biexponential function yielded a blood flow parameter of 95.6 +/- 10.9 ml/min/100 g tissue weight for the brain and 108.0 +/- 13.1 ml/min/100 g tissue weight for the heart. The mean muscle blood flow parameter determined by the monoexponential uptake function was 43.8 +/- 7.3 ml/min/100 g tissue weight. CONCLUSIONS: The blood flow parameter measurement by means of an imaging coil for deuterium is less invasive and reflects the mean tissue blood flow parameter for the entire tissue sample more homogeneously than spectroscopic monitoring.

Quantitative measurement of uptake of radiolabeled monoclonal antibody by means of planar data.

Data obtained from planar images were used to measure the uptake of monoclonal antibody in organs and tumors. Background counts included in the region of interest were eliminated, and the attenuation of the photons emitted by the radionuclide through the intervening tissues was compensated for. The background counts and the intensity of the attenuation were determined from the results of phantom studies and numerical integration with a personal computer. The quantitative uptake of 111In labeled anti-melanoma Monoclonal Antibody (ZME-018) in a melanoma lesion, the liver, and the bone marrow of a patient was measured by the planar method which we developed.

Serial water changes in human skeletal muscles on exercise studied with proton magnetic resonance spectroscopy and imaging.

In vivo 1H-magnetic resonance imaging (MRI) enabled us to study the distribution of water in living tissues and to document changes in human skeletal muscles during physical exercise. The purpose of the present study was to determine the total muscle water changes after exercise using water in 1H-MR spectroscopy and to compare these changes to the signal intensity change on T2*-weighted images and/or to the T2 value change. Seven young male volunteers were positioned in a 1.5 T Philips MR imaging system. They were then asked to dorsiflex their ankle joint against a 2 kg weight once every 2 seconds for 2 minutes. The peak height of water declined according to the clearance curve after exercise in all seven cases with the 1H-MRS similar to the signal intensity. The increasing rate at peak height of total muscle water exceeded both the signal intensity and the T2 value because the water peak height on the 1H-MRS included the extracellular water. In addition, we measured the changes in signal intensity in both calf muscles after walking race exercise. The time intensity curves were used to draw a clearance curve for each muscle group after exercise. It was possible to discern which muscle was used most from the T2*-weighted image that was obtained once after exercise.

Evaluation of blood flow patterns of true and false lumens in dissecting aneurysms using MR phase-contrast techniques.

The magnetic resonance phase-contrast technique for the measurement of flow velocity and volume in true and false lumens was studied in six patients with chronic dissecting aneurysms. Phase-contrast images were obtained at a level perpendicular to the dissecting aneurysms of the descending aorta. As the maximum diameter of aneurysms increased, the ratio of the cross-sectional area of the false to the true lumen increased and the peak average velocity in the true lumen during systole was decreased. This technique proved invaluable for determining prognosis and operability for this condition.

The functional staging of Duchenne muscular dystrophy using in vivo 31P MR spectroscopy.

We applied 31P magnetic resonance spectroscopy (MRS) for the diagnosis of Duchenne muscular dystrophy (DMD), and evaluated its suitability for early-state DMD diagnosis. The spectral patterns showed typical differences between normal and diseased calf muscle. The signal intensity from each spectrum became weaker with advancing disease stage. We could detect abnormality even at the before imaging modalities.