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Purpose The current practice of calculating the specific absorption rate (SAR) relies on local temperature measurements made using temperature probes. For an accurate SAR measurement, a temperature imaging method that provides high temperature sensitivity is desirable, because acceptable levels of SAR produce small temperature changes. MR thermometry using paramagnetic lanthanide complexes can be used to obtain absolute temperature measurements with sub-degree temperature and sub-millimeter spatial resolution. The aim of this study was to develop and evaluate a high temperature resolution MR technique to determine SAR. Methods MR thermometry using a paramagnetic lanthanide complex thulium 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrakis (methylene phosphonate) (TmDOTP5−), which has an almost 102 times stronger chemical shift temperature dependence than water, was used to develop a novel method for SAR measurement. Three-dimensional temperature and SAR images were calculated using MR images acquired with a conventional gradient recalled echo sequence and SAR-intensive T1ρ sequence. Effects of the presence of conducting wire and increasing T1ρ spin-lock pulse duration were also examined. Results SAR distribution could be visualized clearly and surges associated with conducting wires and increasing pulse duration were identified clearly in the computed high spatial resolution SAR images. Conclusion A novel method with high temperature sensitivity is proposed as a tool to evaluate radiofrequency safety in MRI. Magn Reson Med, 2015. © 2015 Wiley Periodicals, Inc.

Transmembrane sodium (23Na) gradient is critical for cell survival and viability and a target for the development of anti-cancer drugs and treatment as it serves as a signal transducer. The ability to integrate abdominal 23Na MRI in clinical settings would be useful to non-invasively detect and diagnose a number of diseases in various organ systems. Our goal in this work was to enhance the quality of 23Na MRI of the abdomen using a 3-Tesla MR scanner and a novel 8-channel phased-array dual-tuned 23Na and 1H transmit (Tx)/receive (Rx) coil specially designed to image a large abdomen region with relatively high SNR. A modified GRE imaging sequence was optimized for 23Na MRI to obtain the best possible combination of SNR, spatial resolution, and scan time in phantoms as well as volunteers. Tissue sodium concentration (TSC) of the whole abdomen was calculated from the inhomogeneity-corrected 23Na MRI for absolute quantification. In addition, in vivo reproducibility and reliability of TSC measurements from 23Na MRI was evaluated in normal volunteers. 23Na axial images of the entire abdomen with a high spatial resolution (0.3 cm) and SNR (~20) in 15 min using the novel 8-channel dual-tuned 23Na and 1H transmit/receive coil were obtained. Quantitative analysis of the sodium images estimated a mean TSC of the liver to be 20.13 mM in healthy volunteers. Our results have shown that it is feasible to obtain high-resolution 23Na images using a multi-channel surface coil with good SNR in clinically acceptable scan times in clinical practice for various body applications.

Although phosphorus-31 (³¹P) magnetic resonance spectroscopy holds potential as noninvasive tool to monitor treatment response of liver malignancies, the lack of appropriate coils has so far restricted its use to liver lesions close to the surface. A novel eight-channel phased-array dual-tuned ³¹P/¹H coil that can assess ³¹P metabolism in deeper liver tissue as well is presented in this article. Analysis of its performance demonstrates that this coil can provide good sensitivity across a width of 20 cm, thereby enabling magnetic resonance spectroscopic imaging (MRSI) scans that can fully cover axial views of the abdomen in lean subjects. In vivo results and reproducibility of ³¹P MRSI at 3 T of axial slices covering the full depth of the liver are shown in healthy volunteers. To minimize intrasubject and intersubject data variability, spectra are corrected for coil sensitivities. Methods to maximize the reproducibility of coil placement and spectroscopic planning are discussed. The phosphomonoesters/phosphodiesters ratio calculated in healthy volunteers has an average intrasubject variation of 23% averaged over voxels selected from the entire liver. Finally, the feasibility of using the coil in the clinic is shown by preliminary ³¹P liver MRSI data obtained from a patient with hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) and liver metastases are an increasing problem worldwide. Non-invasive methods for the early detection of HCC and understanding of the tumor growth mechanisms are highly desirable. Both the diffusion-weighted 1H (DWI) and 23Na MRI reflect alterations in tissue compartment volumes in tumors, as well as physiological and metabolic transformation in cells. Effects of untreated growth on apparent diffusion coefficient of water (ADC), single quantum (SQ) and triple quantum-filtered (TQF) 23Na MRI were compared in intrahepatically and subcutaneously implanted HCCs in rats. Animals were examined weekly for 4 weeks after injection of N1S1 cells. ADC of intrahepatic HCC was 1.5-times higher compared to the nearby liver tissue, and with growth, the ADC did not increase. ADC of subcutaneous HCC was lower compared to intrahepatic HCC and it increased with growth. Untreated growth of both intrahepatic and subcutaneous HCCs was associated with an increase in SQ and TQF 23Na signal intensity suggesting an increase in tissue Na+ and intracellular Na+ (Na+i), respectively, most likely due to an increase in relative extracellular space and Na+i concentration as a result of changes in tissue structure and cellular metabolism. Thus, SQ and TQF 23Na MRI may be complementary to diffusion imaging in areas susceptible to motion for characterizing hepatic tumors and for other applications, such as, predicting and monitoring therapy response. Copyright © 2011 John Wiley & Sons, Ltd.

A magnetic resonance (MR) technique is developed to produce controlled radio-frequency (RF) hyperthermia (HT) in subcutaneously-implanted 9L-gliosarcoma in Fisher rats using an MR scanner and its components; the scanner is also simultaneously used to monitor the tumour temperature and the metabolic response of the tumour to the therapy. The method uses the (1)H chemical shift of thulium 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra-acetic acid (TmDOTA(-)) to monitor temperature. The desired HT temperature is achieved and maintained using a feedback loop mechanism that uses a proportional-integral-derivative controller. The RF HT technique was able to heat the tumour from 33 degrees to 45 degrees C in approximately 10 min and was able to maintain the tumour temperature within +/-0.2 degrees C of the target temperature (45 degrees C). Simultaneous monitoring of the metabolic changes with RF HT showed increases in total tissue and intracellular Na(+) as measured by single-quantum and triple-quantum filtered (23)Na MR spectroscopy (MRS), respectively, and decreases in intra- and extracellular pH and cellular bioenergetics as measured by (31)P MRS. Monitoring of metabolic response in addition to the tumour temperature measurements may serve as a more reliable and early indicator of therapy response. In addition, such measurements during HT treatment will enhance our understanding of the tumour response mechanisms during HT, which may prove valuable in designing methods to improve therapeutic efficiency.

MR thermometry based on the water (1)H signal provides high temporal and spatial resolution, but it has low temperature sensitivity (approximately 0.01 ppm/degrees C) and requires monitoring of another weaker signal for absolute temperature measurements. The use of the paramagnetic lanthanide complex, thulium 1,4,7,10- tetraazacyclododecane-1,4,7,10-tetramethyl-1,4,7,10-tetraacetate (TmDOTMA(-)), which is approximately 60 times more sensitive to temperature than the water (1)H signal, is advanced to image absolute temperatures in vivo using water signal as a reference. The temperature imaging technique was developed using gradient echo and asymmetric spin echo imaging sequences on 9.4 Tesla (T) horizontal and vertical MR scanners. A comparison of regional temperatures measured with TmDOTMA(-) and fiber-optic probes showed that the accuracy of imaging temperature is

The review summarizes the author’s results and literature data on the evaluation of diffusion-weighted magnetic resonance imaging (DWI) as a cancer biomarkers that reflects structural, cellular, apoptotic, and necrotic changes in tumor tissue. Diffusion measurements reflect the effective displacement of water molecules allowed to migrate for a given time. It was demonstrated that apparent diffusion coefficient (ADC) of water estimated from 1H DWI is important tool for the detection and characterization of neoplastic transformation as well as monitoring response to therapy. The possible mechanisms of pre- and post-therapy changes in water ADC in animal body tumors are discussed.

The mechanism of water and sodium apparent diffusion coefficient (ADC) changes in rat skeletal muscle during global ischemia was examined by in vivo 1H and 23Na magnetic resonance spectroscopy (MRS). The ADCs of Na+ and water are expected to have similar characteristics because sodium is present as an aqua-cation in tissue. The shift reagent, TmDOTP5(-), was used to separate intra- and extracellular sodium (Na+i and Na+e, respectively) signals. Water, total tissue sodium (Na+t), Na+i, and Na+e ADCs were measured before and 1, 2, 3, and 4 hr after ischemia. Contrary to the general perception, Na+i and Na+e ADCs were identical before ischemia. Thus, ischemia-induced changes in Na+e ADC cannot be explained by a simple change in the size of relative intracellular or extracellular space. Na+t and Na+e ADCs decreased after 2-4 hr of ischemia, while water and Na+i ADC remained unchanged. The correlation between Na+t and Na+e ADCs was observed because of high Na+e concentration. Similarly, the correlation between water and Na+i ADCs was observed because cells occupy 80% of the tissue space in the skeletal muscle. Ischemia also caused an increase in the Na+i and an equal decrease in Na+e signal intensity due to cessation of Na+/K+-ATPase function.

Purpose To examine the effects of the alkylating anticancer drug 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) on 23Na MRI and the water apparent diffusion coefficient (ADC) in subcutaneously- (sc-) implanted 9L glioma in rats. Materials and Methods 23Na MRI and 1H water ADC measurements were performed on sham-treated control (N = 6) and BCNU-treated (N = 15) Fisher rats one day before BCNU injection and then one, three, and five days after BCNU injection. Results The BCNU-treated tumors were divided into BCNU-responsive (RBCNU) and BCNU-nonresponsive (NRBCNU) groups depending on the tumor volume changes that occurred after therapy. The pretreatment 23Na MRI signal intensity (SI) and water ADC values were higher in RBCNU tumors compared to NRBCNU tumors. 23Na MRI SI and water ADC increased with tumor growth in control and NRBCNU groups, but these changes were interrupted by BCNU therapy in RBCNU group. Conclusion 23Na MRI and water ADC measurements may be useful for predicting and monitoring response to chemotherapy in some tumors. However, the changes that occurred in 23Na MRI SI and water ADC in sc-implanted 9L tumors are in contrast to previously published results for BCNU therapy of orthotopic 9L tumors. This may have important implications for monitoring therapy response in tumors. J. Magn. Reson. Imaging 2006. © 2006 Wiley-Liss, Inc.

Non-invasive thermometry using hyperfine-shifted MR signals from paramagnetic lanthanide complexes has attracted attention recently because the chemical shifts of these complexes are many times more sensitive to temperature than the water 1H signal. Among all the lanthanide complexes examined thus far, thulium tetramethyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (TmDOTMA−) appears to be the most suitable for MR thermometry. In this paper, the feasibility of imaging the methyl 1H signal from TmDOTMA− using a frequency-selective radiofrequency excitation pulse and chemical shift-selective (CHESS) water suppression is demonstrated. A temperature imaging method using a phase-sensitive spin-echo imaging sequence was validated in phantom experiments. A comparison of regional temperature changes measured with fiber-optic probes and the temperatures calculated from the phase shift near each probe showed that the accuracy of imaging the temperature with TmDOTMA− is at least 0.1–0.2°C. The feasibility of imaging temperature changes in an intact rat at 0.5–0.6 mmol/kg dose in only a few minutes is demonstrated. Similar to commonly used MRI contrast agents, the lanthanide complex does not cross the blood–brain barrier. TmDOTMA− may prove useful for temperature imaging in many biomedical applications but further studies relating to acceptable dose and signal-to-noise ratio are necessary before clinical applications. Copyright © 2006 John Wiley & Sons, Ltd.

The acute effects of hyperthermia on intracellular Na+ (Nai+), bioenergetic status and intracellular pH (pHi) were investigated in superfused Radiation Induced Fibrosarcoma-1 (RIF-1) tumour cells using shift-reagent-aided 23Na and 31P nuclear magnetic resonance (NMR) spectroscopy. Hyperthermia at 45 degrees C for 30 min produced a 50% increase in Na, a 0.42 unit decrease in pHi and a 40-45% decrease in NTP/P(i). During post-hyperthermia superfusion at 37 degrees C, pHi and NTP/P(i) recovered to the baseline value, but Na initially decreased and then increased to the hyperthermic level 60 min after heating. Hyperthermia at 42 degrees C caused only a 15-20% increase in Nai+. In the presence of 3 microM 5-(N-ethyl-N-isopropyl)amiloride (EIPA), an inhibitor of the Na+/H+ exchanger, the increase in Nai+ during 45 degrees C hyperthermia was attenuated, suggesting that the heat-induced increase in Nai+ was mainly due to an increase in Na+/H+ anti-porter activity. EIPA did not prevent hyperthermia-induced acidification. This suggests that pHi is controlled by other ion exchange mechanisms in addition to the Na+/H+ exchanger. EIPA increased the thermo-sensitivity of the RIF-1 tumour cells only slightly as measured by cell viability and clonogenic assays. The hyperthermia-induced irreversible increase in Nai+ suggests that changes in transmembrane ion gradients play an important role in cell damage induced by hyperthermia.

Noninvasive techniques to monitor temperature have numerous useful biomedical applications. However, MR thermometry techniques based on the chemical shift, relaxation rates, and molecular diffusion rate of the water 1H signal suffer from poor thermal resolution. The feasibility of MR thermometry based on the strong temperature dependence of the hyperfine-shifted 1H signal from the paramagnetic lanthanide complex thulium-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (TmDOTA-) was recently demonstrated. The use of paramagnetic lanthanide complexes for MR thermometry can be further enhanced by improving the signal-to-noise ratio (SNR) of the observed signal. In this study, the use of lanthanide complexes of a methyl-substituted analog of DOTA4-, 1,4,7,10-tetramethyl 1,4,7,10-tetra azacyclodoecane-1,4,7,10-tetraacetic acetate (DOTMA4-) was evaluated. DOTMA4- complexes have 12 magnetically equivalent methyl protons, which provide an intense and sharper resonance compared to the corresponding DOTA- complexes. Experiments with paramagnetic Pr3+, Yb3+, Tb3+, Dy3+, and Tm3+ complexes of DOTMA4- showed that the Tm3+ complex is most favorable for MR thermometery because of the high temperature dependence of its chemical shift and its relatively narrow linewidth. The chemical shift of the methyl 1H signal from TmDOTMA- was approximately 60 times more sensitive to temperature than the water 1H shift and was insensitive to changes in concentration, pH, [Ca2+], or the presence of other ions and macromolecules. The application of TmDOTMA- for measuring temperature in a subcutaneously implanted tumor model was demonstrated. Lastly, the feasibility of obtaining 3D images from the methyl 1H resonance of TmDOTMA- was demonstrated in phantom and live animal experiments. Overall, TmDOTMA- appears to be a promising probe for MR thermometry in vivo.

Most perfused tumor cell experiments are performed at 37 degrees C, the normal healthy body temperature. However, the temperature of subcutaneously implanted tumors in small animals is generally 29-33 degrees C when the rectal temperature of the animal is maintained at 37 degrees C. We have investigated the acute effects of increasing the temperature of perfused radiation-induced-fibrosarcoma (RIF-1) tumor cells from 33 to 37 degrees C (30 min) on intracellular sodium (Na(i)+) , intracellular pH (pH(i)), and bioenergetic status. Heating the cells by 4 degrees C produced a reversible increase in Na(i)+, slight acidification and no change in nucleotide triphosphate to inorganic phosphate ratio (NTP/P(i)) as measured by shift-reagent-aided (23)Na and (31)P NMR spectroscopy. In the presence of 3 microM 5-(N-ethyl-N-isopropyl) amiloride (EIPA), a potent and specific inhibitor of Na(+)/H(+) antiporter, the increase in Na(i)+ during the heating was completely abolished suggesting that the heat induced increase in Na(i)+ was caused by an increase in Na(+)/H(+) antiporter activity. However, the changes in pH(i) with the heating were identical with or without EIPA, indicating that pH(i) is controlled by other ion exchange mechanisms in addition to Na(+)/H(+) antiporter. NTP/P(i) was significantly higher in presence of EIPA for some time points during the heating suggesting that both NTP production and consumption rates may be altered during the heating. These results indicate that a slight increase in temperature from 33 to 37 degrees C induces significant changes in Na(+) physiology largely because of activation of Na(+)/H(+) antiporter but other ion exchange mechanisms are also involved in maintaining pH(i) in the RIF-1 tumor cells. Thus, care must be taken in choosing the temperature for perfused cell studies.

The purpose of this study was to develop a reliable, noninvasive method for early detection of tumor response to therapy that would facilitate optimization of treatment regimens to the needs of the individual patient. In the present study, the effects of cyclophosphamide (Cp, a widely used alkylating agent) were monitored in a murine radiation induced fibrosarcoma (RIF-1) using in vivo (1)H NMR spectroscopy and imaging to evaluate the potential of these techniques towards early detection of treatment response. Steady-state lactate levels and Gd-DTPA uptake kinetics were measured using selective multiple quantum coherence (Sel-MQC) transfer spectroscopy and dynamic contrast enhanced imaging, respectively in RIF-1 tumors before, 24 and 72 h after 300 mg/kg of Cp administration. High-resolution (1)H NMR spectra of perchloric acid extracts of the tumor were correlated with lactate and glucose concentrations determined enzymatically. In vivo NMR experiments showed a decrease in steady-state lactate to water ratios (5.4 +/- 1.6 to 0.6 +/- 0.5, p < 0.05) and an increase in Gd-DTPA uptake kinetics following treatment response. The data indicate that decreases in lactate result from decreased glycolytic metabolism and an increase in tumor perfusion/permeability. Perchloric acid extracts confirmed the lower lactate levels seen in vivo in treated tumors and also indicated a higher glycerophosphocholine/phosphocholine (GPC/PC) integrated intensity ratio (1.39 +/- 0.09 vs 0.97 +/- 0.04, p < 0.01), indicative of increased membrane degradation following Cp treatment. Steady-state lactate levels provide metabolic information that correlates with changes in tumor physiology measured by Gd-DTPA uptake kinetics with high spatial and temporal resolution. Both of these parameters may be useful for monitoring early tumor response to therapy.

Non-invasive thermometry is pivotal to the future advances of regional hyperthermia as a cancer treatment modality. Current magnetic resonance (MR) thermometry methods suffer from poor thermal resolution due to relatively weak dependence of chemical shift of the (1)H water signal on temperature. This study evaluated the feasibility of using thulium-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (TmDOTA(-)) for MR thermometry. TmDOTA(-) is non-toxic and the gadolinium complex of DOTA(4-) is widely used as a MR contrast agent. The results demonstrate that the temperature dependence of the TmDOTA(-) proton shifts are about two orders of magnitudes higher than the water proton and, thus, provide excellent accuracy and resolution. In addition, TmDOTA(-) proton shifts are insensitive to the paramagnetic complex concentration, pH, Ca(2+) or presence of plasma macromolecules and ions. Because hyperthermia is known to produce changes in tissue pH and other physiological parameters, these properties of TmDOTA(-) greatly simplify the procedures for using the lanthanide complex for MR thermometry. Application of TmDOTA(-) for measurement of temperature in a subcutaneously implanted human melanoma xenograft is demonstrated. Finally, the feasibility of imaging one of the (1)H resonances of the lanthanide complex is demonstrated in phantom experiments. Overall, TmDOTA(-) appears to be a promising probe for MR thermometry in vivo.

The utility of triple-quantum (TQ)-filtered 23 Na NMR spectroscopy for discriminating between intra- and extracellular Na + (Na i + and Na e + , respectively) in a solid tumor in vivo was evaluated using TmDOTP 5− as a 23 Na shift reagent. Infusion of 80 mM TmDOTP 5− without added Ca 2+ produced baseline-resolved Na i + and Na e + peaks in both single-quantum (SQ) and TQ-filtered 23 Na spectra. The Na i + signal represented 22±4% of the SQ spectrum, but 59±10% of the TQ-filtered spectrum. Therefore, the Na i + contribution in TQ-filtered spectra is much higher than in SQ spectra. Both SQ and TQ-filtered Na i + signals increased by about 75% 1 h after sacrificing the animal. The TQ-filtered relaxation times did not change during this time, indicating that changes observed in TQ-filtered spectra collected with a preparation time of 3 ms represent changes in the concentration of sodium ions contributing to the TQ-filtered signal. Similar experiments were conducted without TmDOTP 5− to determine changes in the Na e + signal in the absence of the shift reagent. The changes in total SQ and TQ-filtered signals 1 h after sacrificing the animal showed that the SQ Na e + signal decreased by approximately 35%, while the TQ-filtered Na e + signal did not change significantly. This demonstrates that the TQ-filtered 23 Na signal is relatively insensitive to changes in Na e + content. To our knowledge, this work represents the first evaluation of multiple-quantum-filtered 23 Na spectroscopy to discriminate between intra- and extracellular Na + in a solid tumor in vivo .

Rationale and Objectives The authors performed this study to evaluate the selective acidification of a human melanoma xenograft in mice with severe combined immunodeficiency with the induction of hyperglycemia (mean blood glucose level ± standard error of the mean, 26 mmol/L ± 1) and the intraperitoneal administration of metaiodobenzylguanidine (MIBG, 30 mg/kg), α-cyano-4-hydroxycinnamate (CNCn, 300 mg/kg), lonidamine (100 mg/kg), cariporide (HOE642, 160 mg/kg), or 4,4′-diisothiocyanatostilbene-2, 2'-disulfonic acid (DIDS, 50 mg/kg). Materials and Methods The intra- and extracellular pH levels of tumor were estimated from the chemical shifts of inorganic phosphate and 3-aminopropylphosphonate, respectively, with phosphorus-31 nuclear magnetic resonance (MR) spectroscopy. The relative level of steady-state lactate was monitored with hydrogen-1 MR spectroscopy. Results In small tumors (≤8.0 mm), hyperglycemia decreased the intra- and extracellular pH levels by less than 0.2. The combination of hyperglycemia and MIBG decreased the intra- and extracellular pH levels by approximately 0.4 and 0.6, respectively, and lowered the β-nucleoside triphosphate (NTP)/inorganic phosphate (P i ) ratio of tumor and liver by about 60% and 25%, respectively. The combination of hyperglycemia, MIBG, and CNCn produced a transient decrease in the intracellular pH of about 0.6. The combination of hyperglycemia and lonidamine produced a sustained (>3 hours) 0.8-unit decrease in intracellular pH and an 83% and 100% decrease in PCr/P i and β-NTP/P i ratios, respectively. The combination of hyperglycemia, MIBG, cariporide, and DIDS produced a gradual decrease in intra- and extracellular pH by 1.1 and 1.0, respectively. The relative level of steady-state lactate concentration in tumors increased 10% with hyperglycemia alone, about 20% with MIBG plus hyperglycemia, and increased more than twofold when hyperglycemia was combined with MIBG and CNCn administration. Conclusion These preliminary data suggest that hyperglycemia and combinations of respiratory and ion transport inhibitors can be used to selectively acidify tumors and, thereby, sensitize them to hyperthermia or other pH-sensitive therapeutic modalities.

The use of TmDOTP(5-) as an in vivo (23)Na NMR shift reagent (SR) for subcutaneously implanted 9L gliosarcoma was evaluated. TmDOTP(5-) produced a single sharp extracellular peak after about 50-60 min of infusion, and did not cause any changes in the (31)P resonance areas or chemical shifts, suggesting that the SR is homogeneously distributed in the extracellular space and does not alter tumor bioenergetic status. TmDOTP(5-) and CoEDTA(-) as extracellular space markers gave identical results for relative extracellular space (0.25 +/- 0.03 and 0.25 +/- 0.04, respectively) and intracellular Na(+) concentration (19.3 +/- 4.0 mM and 18.6 +/- 3.9 mM, respectively), indicating that the biodistribution of the SR is the same as the well-accepted extracellular space marker. The in vivo T(1) and T(2) relaxation times of intra- and extracellular Na(+) were also measured. Our results indicate that TmDOTP(5-) promises to be an effective shift reagent and extracellular space marker in the 9L gliosarcoma and perhaps other tumors. Magn Reson Med 45:436-442, 2001.

The acute affect of voluntary muscle contractions performed by healthy volunteers was evaluated using (23)Na nuclear magnetic resonance (NMR). Three-dimensional gradient-echo (23)Na images, pulse-acquired spectra, and transverse relaxation times were obtained before and after ankle flexion-extension exercise. The muscle sodium concentration was calculated from (23)Na images using a 40 mM NaCl standard and the measured T(2) values. Before exercise the muscle sodium concentration was 26+/-4 mmole/kg wet weight. This agrees closely with literature values, suggesting that muscle Na(+) is fully NMR visible. The (23)Na image intensity increased by 34%+/-7% in the exercised muscle and diminished with a half-life of 30+/-6 minutes. The pulse-acquired spectra, however, did not show any significant change in muscle signal intensity following exercise, but the relative contribution of the slow T(2) component increased. The calculated sodium concentration also did not change significantly after the exercise. We therefore infer that the changes in (23)Na magnetic resonance imaging (MRI) were due to a change in sodium-macromolecular interaction rather than a change in tissue sodium content. We believe that this report represents the first study of (23)Na MRI of skeletal muscle.

The application of continuous wavelet transform (CWT) analysis technique is presented to analyze multiple-quantum-filtered (MQF) 23 Na magnetic resonance spectroscopy (MRS) data. CWT acts on the free-induction-decay (FID) signal as a time-frequency variable filter. The signal-to-noise ratio (SNR) and frequency resolution of the output filter are locally increased. As a result, MQF equilibrium longitudinal magnetization and the apparent fast and slow transverse relaxation times are accurately estimated. A developed iterative algorithm based on frequency signal detection and components extraction, already proposed, was used to estimate the values of the signal parameters by analyzing simulated time-domain MQF signals and data from an agarose gel. The results obtained were compared to those obtained by measurement of signal height in frequency domain as a function of MQF preparation time and those obtained by a simple time-domain curve fitting. The comparison indicates that the CWT approach provides better results than the other tested methods that are generally used for MQF 23 Na MRS data analysis, especially when the SNR is low. The mean error on the estimated values of the amplitude signal and the apparent fast and slow transverse relaxation times for the simulated data were 2.19, 6.63, and 16.17% for CWT, signal height in frequency domain, and time-domain curve fitting methods, respectively. Another major advantage of the proposed technique is that it allows quantification of MQF 23 Na signal from a single FID and, thus, reduces the experiment time dramatically.

Isolated, perfused rat livers were examined by single-quantum (SQ) and double-quantum-filtered (DQ-filtered) 23Na spectroscopy during prolonged global ischemia and during perfusion with ouabain, low-buffer potassium, or lithium-enriched buffer. Baseline separation of the intracellular (Na(i)+) and extracellular (Na(e)+) sodium resonances using TmDOTP5- allowed a direct comparison of temporal changes in SQ versus DQ-filtered Na(i)+. The SQ Na(i)+ signal increased approximately 150% during the first 15 min of global ischemia and then remained relatively constant over the next 45 min, while the DQ-filtered signal steadily increased approximately 400% over the same 60 min period. In similar experiments in which all perfusate sodium was replaced by lithium, the DQ-filtered Na(i)+ signal increased approximately 180% over a similar period of ischemia. Exposure of livers to ouabain also resulted in larger increases in DQ-filtered versus SQ signal of Na(i)+. The approximately 290% increase in DQ-filtered sodium observed during perfusion of livers with a hypokalemic buffer (1.2 mM K+) could be completely reversed by continued perfusion with a buffer containing normal levels of K+ (4.7 mM). These data suggest that the DQ-filtered Na(i)+ signal of liver does not simply report an increase in [Na(i)+], but may be exquisitely sensitive to other intracellular events initiated by altered physiology.

Multiple-quantum (MQ) -filtered 23Na NMR has been proposed as a means to partially discriminate between intracellular and extracellular sodium. However, low signal-to-noise ratio (SNR) has been a major obstacle to MQ-filtered 23Na imaging becoming an important technique for biological and clinical applications. We compared the various MQ-filtered 23Na imaging pulse sequences to select the optimum sequence that provides the best SNR. The results of phantom experiments show that the gradient-echo MQ-filtered 23Na imaging sequence produces the best SNR. We also report, for the first time, three-dimensional single-quantum (SQ) and triple-quantum (TQ) -filtered 23Na images of the live dog brain and demonstrate the sensitivity of these images to ischemia produced by euthanizing the animal. The SQ images showed a 10% to 15% decrease in signal intensity from the brain post-mortem, whereas the TQ-filtered images showed a 40% to 50% increase. These changes in signal intensities are consistent with the influx of Na+ into the cells upon death. The feasibility of obtaining TQ-filtered 23Na images of in situ dog brain encourages us to apply this technique to humans.

Anisotropic nuclear quadrupole interactions can produce residual quadrupole splitting in the NMR spectra of rapidly moving quadrupolar nuclei in incompletely disordered aqueous heterogeneous systems. Such systems may include hydrated sodium nuclei in biological tissue and biopolymer gels. To describe the NMR signals from such samples, we use a domain model in which each domain is characterized by a quadrupole frequency and a residence time of the nucleus. We show that the signals from each domain after one pulse, the quadrupole echo sequence, and the various multiple quantum filters (MQFs) can be expressed as a linear combination of five different phase coherences. To simulate the effect of various distributions (Pake powder pattern, Gaussian, etc.) of quadrupole frequencies for different domains on the NMR signal, we have written the computer program CORVUS. CORVUS also includes the effects of exchange between different domains using diffusion and random jump models. The results of computer simulations show that the Gaussian and Pake powder pattern quadrupole frequency distributions produce very different phase coherences and observable NMR signals when the exchange rate (1/taue) between different domains is slow. When 1/taue is similar to the root mean square quadrupole frequency (final sigma), the signals from the two distributions are similar. When 1/taue is an order of magnitude greater than final sigma, there is no apparent evidence of quadrupole splitting in the shape of the signal following one pulse, but the residual effects of the quadrupole splitting make a significant contribution to the fast transverse relaxation rate. Therefore, in this case, it is inappropriate to use the observed biexponential relaxation rates to obtain a single correlation time. The quadrupole echo and the various MQF signals contain an echo from the satellite transitions in the presence of quadrupole splitting. The peak of this echo is very sensitive to 1/taue. The time domain analysis of these signals is more direct and less ambiguous than the frequency domain analysis because the echo does not occur at the beginning of data acquisition. The quadrupole echo pulse sequence is the most sensitive detector of residual quadrupole splitting and exchange of sodium ions between different domains. However, if the sample is compartmentalized so that only a fraction of the nuclei have quadrupole splitting, the double quantum magic angle filter (DQ-MA) is more suitable. This is because the DQ-MA signal contains only the contributions from satellite transitions. Use of simulations to analyze signals from various one-pulse, quadrupole echo, and multiple quantum filter pulse sequences can yield information on substrate order and aid in quantitation of multiple quantum filter signals.

Low signal-to-noise ratio (SNR) has been the main obstacle to multiple quantum-filtered 23Na imaging becoming an important technique for biologic and clinical applications. Through computer simulations and phantom experiments, we show that the SNR in 23Na imaging can be substantially improved by weighted signal averaging. Three-dimensional single quantum and triple quantum (TQ)-filtered 23Na images of an externalized rabbit kidney were collected with this technique. The TQ-filtered image did not show any signal when the animal was alive. However, upon sacrificing the animal, the renal cortex became clearly visible without any significant increase in signal from the medullary region. This increase in TQ-filtered signal in the renal cortex may be caused by an increased concentration of intracellular Na+ in the large intracellular space present herein, compared with the medulla. To our knowledge, this study represents the first example of three-dimensional TQ-filtered 23Na image of a biological sample.

Since transmembrane sodium gradient is essential to man cell functions, there is continuing interest in methods tha differentiate intracellular and extracellular Na + . In the kidney, shift reagent (SR) aided 23 Na magnetic resonance spectros copy (MRS) has been successfully used only in isolated cells tubules, and the perfused organ. In this report, we demon strate for the first time that TmDOTP 5- can be used to distin guish Na + compartments in kidneys in vivo. Infusion of 80 mM TmDOTP 5- without added Ca 2+ produced three resolved 23 Na resonances, which we have assigned to intracellular Na + , vascular Na + , and intraluminal Na + . In comparison, infusion o 400 mM DyTTHA 3- produced two broad and unresolved resonances. The 31 P spectra of the cellular high energy phosphate metabolites indicate that TmDOTP 5- is safe for in vivo applications. Washout studies suggest that this SR display renal clearance similar to that of MR imaging contrast agents However, the glomerular filtration rate (GFR) in animals in fused with TmDOTP 5- was reduced by 49% compared with the GFR in control animals, perhaps due to the hypotensive effects of the SR. We conclude that TmDOTP 5- is effectively cleared from the blood of live animals but that a different formulation will be required for clinical application.

The cellular events leading to the proliferation of new preretinal blood vessels into the vitreous are unknown. It has been hypothesized that a metabolic trigger, such as increased lactate concentration, induces ambient growth factors to initiate this neovascularization. However, this hypothesis has not been tested in vivo because non-invasive methods for measuring vitreous lactate concentration have not been available. In this study, we identified lactate as the dominant resonance in normal rabbit vitreous using a water-suppressed, localized proton NMR method; negligible spectral contamination from lipid, alanine and threonine was demonstrated. The steady-state vitreous lactate concentration in vivo was 11.3 +/- 4 mM (mean +/- SD, n = 7 eyes) which was not statistically different (p = 0.498) to the enzymatically determined lactate content in vitreous from a parallel group of animals [12.1 +/- 0.6 mM (n = 6 eyes)]. Non-invasive measurement of steady-state vitreous lactate concentration using localized proton NMR represents the necessary first step in understanding the role of lactate in neovascularization.

In vivo 31P- and 23Na-magnetic resonance spectroscopy was used to measure phosphorus metabolites, intracellular pH, cytosolic free Mg2+, and intracellular Na+ in the liver of rats 24 h after 40% total body surface area full-thickness burn injury. Studies were performed during infusion of thulium (III) 1,4,7,10-tetraazacyclododecane N,N′,N″,N″′-tetra(methylenephosphonate), which served as the Na+ shift agent. Compared with the sham-burn group, there was a significant increase in hepatic intracellular Na+ along with a decrease in intracellular pH and free Mg2+. The ratio of intra- to extra-cellular Na+ increased, indicating a decreased Na+ gradient that may determine the hepatic transmembrane potential difference. Hepatic beta-ATP/P(i) also significantly decreased, which suggests that either ATP utilization is significantly accelerated or ATP synthesis is inhibited after the thermal injury. Of the cations measured (Na+, Mg2+, H+), the change in intracellular Na+ was most dramatic. This study demonstrates that major burn injury may cause profound changes in hepatic bioenergetics and ionic metabolism 24 h after injury and that intracellular Na+ may be a sensitive indicator of hepatic dysfunction 24 h after injury. Because these animals tolerated the shift reagent, thuliumIII) 1,4,7,10-tetraazacyclododecane N,N′,N″,N″′-tetra(methylenephosphonate), nuclear magnetic resonance spectroscopy may prove valuable in monitoring intracellular cations in the liver after major injury.

To monitor the effects of the apoptotic agent benzamide riboside (BR) on tumor volume and water apparent diffusion coefficient (ADC) in rat hepatocellular carcinoma (HCC).Water ADC of the tumors and nearby liver tissue was measured using diffusion-weighted 1H MRI (DWI). The two groups of BR-treated animals, which differed in their sensitivity to the treatment, were identified as responsive (RBR) and non-responsive (NRBR).Tumor growth in the RBR group was arrested and the mean tumor volume in this group was 1/6th and 1/16th compared to that of the NRBR group on days 7 and 14 after treatment, respectively. Water ADC of HCC was higher than in nearby normal liver tissue. Before BR treatment, the mean water ADC was significantly higher in the RBR group compared to the NRBR group. BR therapy did not change the water ADC value regardless of tumor sensitivity.Although the water ADC did not change after chemotherapy by BR, DWI has great potential for detecting and predicting response to chemotherapy in HCC.

Although phosphorus-31 (³¹P) magnetic resonance spectroscopy holds potential as noninvasive tool to monitor treatment response of liver malignancies, the lack of appropriate coils has so far restricted its use to liver lesions close to the surface. A novel eight-channel phased-array dual-tuned ³¹P/¹H coil that can assess ³¹P metabolism in deeper liver tissue as well is presented in this article. Analysis of its performance demonstrates that this coil can provide good sensitivity across a width of 20 cm, thereby enabling magnetic resonance spectroscopic imaging (MRSI) scans that can fully cover axial views of the abdomen in lean subjects. In vivo results and reproducibility of ³¹P MRSI at 3 T of axial slices covering the full depth of the liver are shown in healthy volunteers. To minimize intrasubject and intersubject data variability, spectra are corrected for coil sensitivities. Methods to maximize the reproducibility of coil placement and spectroscopic planning are discussed. The phosphomonoesters/phosphodiesters ratio calculated in healthy volunteers has an average intrasubject variation of 23% averaged over voxels selected from the entire liver. Finally, the feasibility of using the coil in the clinic is shown by preliminary ³¹P liver MRSI data obtained from a patient with hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) and liver metastases are an increasing problem worldwide. Non-invasive methods for the early detection of HCC and understanding of the tumor growth mechanisms are highly desirable. Both the diffusion-weighted (1)H (DWI) and (23)Na MRI reflect alterations in tissue compartment volumes in tumors, as well as physiological and metabolic transformation in cells. Effects of untreated growth on apparent diffusion coefficient of water (ADC), single quantum (SQ) and triple quantum-filtered (TQF) (23)Na MRI were compared in intrahepatically and subcutaneously implanted HCCs in rats. Animals were examined weekly for 4 weeks after injection of N1S1 cells. ADC of intrahepatic HCC was 1.5-times higher compared to the nearby liver tissue, and with growth, the ADC did not increase. ADC of subcutaneous HCC was lower compared to intrahepatic HCC and it increased with growth. Untreated growth of both intrahepatic and subcutaneous HCCs was associated with an increase in SQ and TQF (23)Na signal intensity suggesting an increase in tissue Na(+) and intracellular Na(+) (Na(+)(i)), respectively, most likely due to an increase in relative extracellular space and Na(+)(i) concentration as a result of changes in tissue structure and cellular metabolism. Thus, SQ and TQF (23)Na MRI may be complementary to diffusion imaging in areas susceptible to motion for characterizing hepatic tumors and for other applications, such as, predicting and monitoring therapy response.

Hepatocellular carcinoma (HCC) is a common cause of death from solid organ malignancy worldwide. Extracellular signal-regulated/mitogen-activated protein kinase kinase (MEK) signaling is a critical growth regulatory pathway in HCC. Targeting MEK with a novel small molecule inhibitor, PD0325901, may inhibit HCC tumorigenesis. PD0325901 (0.01-100 nM) inhibited growth and MEK activity in vitro in immortalized murine transforming growth factor alpha (TGF-α) transgenic hepatocyte (TAMH) cells, derived from the livers of TGF-α transgenic mice. Treatment of athymic mice bearing TAMH flank tumors with vehicle or PD0325901 (20 mg/kg) revealed a significant reduction of MEK activity ex vivo 24 hours after a single PD0325901 dose. The growth rate of TAMH flank tumors over 16 days was reduced threefold in the treatment arm (1113 ± 269% versus 3077 ± 483%, P < 0.01). PD0325901 exhibited similar inhibitory effects in HepG2 and Hep3B human HCC cells in vitro and in Hep3B flank tumors in vivo. To confirm this in a developmental model, MT-42 (CD-1) TGF-α mice were treated with vehicle or PD0325901 (20 mg/kg) for 5 weeks. Gross HCC was detected in 47% and 13.3% of the control and treatment mice, respectively. Tumor growth suppression by PD0325901 relative to vehicle was also shown by magnetic resonance imaging. These studies provide compelling preclinical evidence that targeting MEK in human clinical trials may be promising for the treatment of HCC. (HEPATOLOGY 2010.)

In vivo sodium-23 and hydrogen-1 magnetic resonance (MR) imaging and spectroscopy of the rat brain during infusion of the shift reagent thulium DOTP5− (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra[methylene phosphonate]) was performed to assign the various peaks observed during infusion and to evaluate the shift reagent in discriminating tissue compartments. Na-23 spectra collected during the infusion showed two shifted peaks that were assigned to intravascular Na+ and extracellular muscle Na+, respectively, and one unshifted peak assigned to intra- and extracellular brain Na+ and cerebrospinal fluid Na+. These assignments were validated with H-1 and Na-23 MR imaging and Na-23 chemical shift imaging (CSI). The H-1 and Na-23 images showed that a surface coil placed on a rat head can detect a substantial amount of signal from muscle surrounding the skull. Na-23 CSI spectra from successive 1-mm-thick coronal sections indicated that the shift reagent did not cross the blood-brain barrier. The study also showed that bulk susceptibility shifts are quite small with Tm-DOTP5−. This reagent may be useful in determining compartmental Na+ concentrations and blood flow kinetics in brain and in examining the integrity of the blood-brain barrier.

Reabsorption of water and other molecules is dependent on the corticomedullary sodium concentration gradient in the kidney. During the early course of acute tubular necrosis (ATN), this gradient is altered. Therefore,23Na magnetic resonance imaging (MRI) was used to study the alterations in renal sodium distribution in the rat kidney during ischemia and reperfusion (IR) injury, which induces ATN. In-magnet ischemia was induced for 0 (control), 10, 20, 30 or 50 min in Wistar rats.23Na images were collected every 10 min during baseline, ischemia, and 60-min reperfusion periods. T1and T2relaxation times were measured by both23Na-MRI and -MRS on a separate cohort of animals during ischemia and reperfusion for correction of relaxation-related tissue sodium concentration (TSC). A marked decrease was observed in the medulla and cortex23Na-MRI signal intensity (SI) during the early evolution of ATN caused by IR injury, with the sodium reabsorption function of the kidney being irreversibly damaged after 50 min of ischemia. Sodium relaxation time characteristics were similar in the medulla and cortex of normal kidney, but significantly decreased with IR. The changes in relaxation times in both compartments were identical; thus the medulla-to-cortex sodium SI ratio represents the TSC ratio of both compartments. The extent of IR damage observed with histological examination correlated with the23Na-MRI data.23Na-MRI has great potential for noninvasive, clinical diagnosis of evolving ATN in the setup of acute renal failure and in differentiating ATN from other causes of renal failure where tubular function is maintained.

MR thermometry based on the water (1)H signal provides high temporal and spatial resolution, but it has low temperature sensitivity (approximately 0.01 ppm/degrees C) and requires monitoring of another weaker signal for absolute temperature measurements. The use of the paramagnetic lanthanide complex, thulium 1,4,7,10- tetraazacyclododecane-1,4,7,10-tetramethyl-1,4,7,10-tetraacetate (TmDOTMA(-)), which is approximately 60 times more sensitive to temperature than the water (1)H signal, is advanced to image absolute temperatures in vivo using water signal as a reference. The temperature imaging technique was developed using gradient echo and asymmetric spin echo imaging sequences on 9.4 Tesla (T) horizontal and vertical MR scanners. A comparison of regional temperatures measured with TmDOTMA(-) and fiber-optic probes showed that the accuracy of imaging temperature is0.3 degrees C. The temperature imaging technique was found to be insensitive to inhomogeneities in the main magnetic field. The feasibility of imaging temperature of intact rats at approximately 1.4 mmol/kg dose with approximately 1-mm spatial resolution in only 3 min is demonstrated. TmDOTMA(-) should prove useful for imaging absolute temperatures in deep-seated organs in numerous biomedical applications.

Leptin is known to be associated with regulation of body weight and fat content. The effects of exogenous leptin on abdominal visceral (VS) and subcutaneous (SC) fat volume and hepatic fat-to-water ratio in leptin-deficient obese mice were investigated by (1)H magnetic resonance imaging (MRI). Chemical shift-selected fat and water (1)H MRI of control and leptin-treated mice were obtained 1 day before treatment and after 7 days of treatment (0.3 mg/kg/day). Hepatic fat-to-water ratio and VS fat volume decreased significantly with treatment, whereas SC fat volume did not change. Noninvasive measurement of fat and water content in different body regions using MRI should prove useful for evaluating new drugs for the treatment of obesity and other metabolic disorders.

Objective Polyethylene glycol (PEG) is a nontoxic molecule with known efficacy as a cell membrane sealant, improving histological and behavioral outcomes in trauma models. Diffusion-weighted (DW) magnetic resonance imaging (MRI) is the most sensitive method of detecting in vivo diffuse axonal injury (DAI), where a decreased apparent diffusion coefficient (ADC) of water reflects cytotoxic edema. We use DW-MRI to assess severe DAI in rats treated with a single acute postinjury injection of PEG. Methods Rats were divided into uninjured, injured saline-treated, and injured PEG-treated groups. Injury groups received a severe brain injury using an impact-acceleration weight-drop model. Saline or PEG was administered acutely as a single intravenous dose to injured saline-treated and injured PEG-treated groups, respectively. DW-MRI analysis was performed at postinjury day 7 with a 9.4-T magnet. ADC was calculated for cortex, corpus callosum/hippocampus, and thalamus in each group. Results An expected decrease in ADC, representing cytotoxic edema, was observed in the injured saline-treated group. The injured PEG-treated group demonstrated no decrease in ADC relative to the uninjured rats, and the difference between ADC in saline and PEG-treated groups reached significance for all 3 zones of assessed brain. Differences were seen grossly between injured saline-treated and injured PEG-treated groups on representative color-mapped ADC images. Conclusion A single intravenous dose of PEG dramatically limits sequelae of severe acceleration-induced brain injury--in this case, assessed by cytotoxic edema on DW-MRI--by intervening at the primary injury level of neuronal membrane disruption. This outcome is unprecedented, as no prior treatments for DAI have demonstrated similar efficacy. DAI treatment with intravenous PEG may have future clinical relevance and warrants further investigation.

The initial results of in vivo 23Na magnetic resonance imaging (MRI) of the human torso at 3-Tesla using an 8-channel dual tuned 23Na and 1H transmit/receive coil for various body applications are presented. We are able to obtain 23Na images of the human torso with 0.3 cm spatial resolution and ∼ 20 SNR in ∼15 min. These images were acquired with imaging parameters optimized under specific absorption rate limit for human scans. Because a trans-membrane sodium gradient is critical for cell survival, the ability to perform 23Na MRI of the torso in clinical settings would be useful to noninvasively detect and diagnose a number of diseases of the liver, gallbladder, pancreas, kidney, spleen and other body organs.

An MR technique has been developed to administer controlled radiofrequency (RF) hyperthermia (HT) to treat sc-implanted tumors using an MR scanner and its components. The method uses the 1H chemical shift of TmDOTA− to monitor tumor temeprature non-invasively. The desired HT temeprature is achieved and maintained using a feedback loop mechanism that uses a proportional-derivative (PD) controller. The RF HT techqinue has been able to heat the tumor from 33 to 45 °C in ∼ 10 min and was able to mainatain the tumor temeprature within ±0.2 °C of the target temperature. Simultaneous monitoring of the metabolic changes with RF HT using multinuclear MRS techniques showed a significant increase in the [Nai]+ as measured by TQF 23Na MRS and a significant decrease in cellular bioenergetics and pH as measured by 31P MRS.

An improvement to the method for imaging with extremely inhomogeneous radio-frequency fields is presented which uses a pseudo-noise-modulated selective (PNMS) excitation prepulse to randomize the signal from outside the selected slice. This prepulse reduces the problems that arise from imperfect subtraction involved with the one-dimensional ISIS-type slice selection procedures. The results demonstrate two to three times improvement in the signal-to-noise ratio and reduction in the image artifacts when this prepulse is used.

OBJECTIVE. The inflammatory subtype of nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, is becoming one of the most important causes of chronic liver disease. In this article, we discuss the epidemiology, pathogenesis, and clinical and radiologic diagnosis of the subtypes of nonalcoholic fatty liver disease.CONCLUSION. We discuss the current and evolving imaging tests in the evaluation of hepatic fatty content, inflammation, and fibrosis.

Increased iron levels and iron-mediated oxidative stress play an important role in the pathogenesis of many neurodegenerative diseases. The finding that mutations in the ferritin light polypeptide (FTL) gene cause a neurodegenerative disease known as neuroferritinopathy or hereditary ferritinopathy (HF) provided a direct connection between abnormal brain iron storage and neurodegeneration. HF is characterized by a severe movement disorder and by the presence of nuclear and cytoplasmic ferritin inclusion bodies in glia and neurons throughout the CNS and in tissues of multiple organ systems. Here we report that the expression in transgenic mice of a humanFTLcDNA carrying a thymidine and cytidine insertion at position 498 (FTL498–499InsTC) leads to the formation of nuclear and cytoplasmic ferritin inclusion bodies. As in HF, ferritin inclusions are seen in glia and neurons throughout the CNS as well as in cells of other organ systems. Our studies show histological, immunohistochemical, and biochemical similarities between ferritin inclusion bodies found in transgenic mice and in individuals with HF. Expression of the transgene in mice leads to a significant decrease in motor performance and a shorter life span, formation of ferritin inclusion bodies, misregulation of iron metabolism, accumulation of ubiquitinated proteins, and incorporation of elements of the proteasome into inclusions. This new transgenic mouse represents a relevant model of HF in which to study the pathways that lead to neurodegeneration in HF, to evaluate the role of iron mismanagement in neurodegenerative disorders, and to evaluate potential therapies for HF and related neurodegenerative diseases.

A novel technique is proposed to facilitate the selective imaging of specific molecules from a mixture. The application of the technique presented here demonstrates the ability to selectively produce 19F MR images of either trifluoroacetic acid or the perfluorocarbon emulsion Oxypherol-ET (perfluorotributylamine), when both molecules are present simultaneously. Selective detection is based on the presence of homonuclear J-modulation in one molecule and differential spin-spin relaxation time (T2). Perfluorotributylamine, an A3B2 system, is subject to homonuclear J-modulation, which produces a null signal from the antiphase components of the triplet (A3) when an echo time (TE) = 1 2J is used in a spin-echo image. At this echo time the second molecule, in this example trifluoroacetic acid, a non-coupled spin system, is selectively imaged. At longer echo times, e.g., TE = 1 J there is substantial recovery of the J-modulated signal, which may be solely observed due to T2 decay of the trifluoroacetic acid signal. The method is demonstrated both using phantoms and in vivo.

In patients with nonalcoholic fatty liver disease, there is not a consistent relationship between severity of steatosis and the presence of steatohepatitis. This leads to the possibility that severity of steatosis is not an important factor in the pathogenesis of nonalcoholic steatohepatitis. Alternatively, it is possible that currently used method to quantify hepatic steatosis (histologic grading) may not accurately reflect hepatic lipid content. Therefore, we examined the relationship between hepatic triglyceride (TG) content and nonbiochemical assessment of steatosis in 38 human liver samples.Hepatic steatosis was histologically graded by the 3 hepatopathologists and hepatic TG levels were determined from liver homogenate. Additionally, we characterized the relationship between (a) hepatic steatosis quantified by magnetic resonance spectroscopy (MRS) and histologic grading and hepatic TG content and (b) hepatic long-chain polyunsaturated fatty acid n-6/n-3 ratio and the severity of steatosis.Twenty-two samples had5% steatosis, 9 had 5% to 33% steatosis and 7 had33% steatosis. The mean (+/-SD) hepatic TG was 1.8+/-1.3 mg/mg of protein and MRS fat score was 6.4+/-6.0. There was a significant correlation between histologic grading and hepatic TG content (r=0.64, P0.001). A significant correlation existed between MRS fat score and histologic grading (r=0.61, P=0.006), and hepatic TG (r=0.63, P=0.004). Severity of steatosis as graded histologically had significant correlation with n-6/n-3 ratio (r=0.61, P0.001).Hepatic steatosis quantified by histologic grading or by MRS is significantly reflective of hepatic TG content. Further research is needed to investigate the relationship we observed between n-6/n-3 ratio and the severity of steatosis.

MR thermometry techniques based on the strong water 1H signal provide high spatial and temporal resolution and have shown promise for applications such as laser surgery and RF ablation. However, these techniques have low temperature sensitivity for hyperthermia applications and are greatly influenced by local motion and susceptibility variations. 1H NMR signals from paramagnetic lanthanide complexes of Pr3+, Yb3+ and Tm3+ show up to 300-fold stronger temperature dependence compared to the water 1H signal. In addition, 1H chemical shifts of many of these complexes are insensitive to other factors such as the concentration of the paramagnetic complex, pH, [Ca2+], and the presence of plasma macro-molecules and ions. Applications of lanthanide complexes for temperature measurement in intact animals and the feasibility of mapping temperatures in phantoms have been demonstrated. Among all the lanthanide complexes examined so far, thulium 1, 4, 7, 10-tetramethyl-1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetate (TmDOTMA-) appears to be the most attractive for in vivo MR thermometry. The 1H signal from the methyl groups on this complex is relatively intense because of 12 equivalent protons and provides high temperature sensitivity because of the large paramagnetic shifts induced by thulium. The possibility of imaging TmDOTMA2--in intact animals at physiologically safe concentrations has recently been demonstrated. Overall, MR thermometry methods based on hyperfine-shifted MR signals from paramagnetic lanthanide complexes appear promising for animal applications, but further studies relating to acceptable dose and signal-to-noise ratio are necessary before clinical use.

Noninvasive techniques to monitor temperature have numerous useful biomedical applications. However, MR thermometry techniques based on the chemical shift, relaxation rates, and molecular diffusion rate of the water 1H signal suffer from poor thermal resolution. The feasibility of MR thermometry based on the strong temperature dependence of the hyperfine-shifted 1H signal from the paramagnetic lanthanide complex thulium-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (TmDOTA-) was recently demonstrated. The use of paramagnetic lanthanide complexes for MR thermometry can be further enhanced by improving the signal-to-noise ratio (SNR) of the observed signal. In this study, the use of lanthanide complexes of a methyl-substituted analog of DOTA4-, 1,4,7,10-tetramethyl 1,4,7,10-tetra azacyclodoecane-1,4,7,10-tetraacetic acetate (DOTMA4-) was evaluated. DOTMA4- complexes have 12 magnetically equivalent methyl protons, which provide an intense and sharper resonance compared to the corresponding DOTA- complexes. Experiments with paramagnetic Pr3+, Yb3+, Tb3+, Dy3+, and Tm3+ complexes of DOTMA4- showed that the Tm3+ complex is most favorable for MR thermometery because of the high temperature dependence of its chemical shift and its relatively narrow linewidth. The chemical shift of the methyl 1H signal from TmDOTMA- was approximately 60 times more sensitive to temperature than the water 1H shift and was insensitive to changes in concentration, pH, [Ca2+], or the presence of other ions and macromolecules. The application of TmDOTMA- for measuring temperature in a subcutaneously implanted tumor model was demonstrated. Lastly, the feasibility of obtaining 3D images from the methyl 1H resonance of TmDOTMA- was demonstrated in phantom and live animal experiments. Overall, TmDOTMA- appears to be a promising probe for MR thermometry in vivo.

Use of the popular club drug ecstasy (3,4-methylenedioxymethamphetamine, MDMA) can result in life-threatening hyperthermia and rhabdomyolysis. Recent studies show a link between skeletal muscle uncoupling proteins in MDMA-mediated hyperthermia. The mechanisms by which MDMA interacts with skeletal muscle mitochondria are largely unknown. The present study was designed to comprehensively evaluate the effects of MDMA on bioenergetics and toxicity of skeletal muscle. Using (31)P nuclear magnetic resonance (NMR) and serum creatine kinase levels, we demonstrate evidence for uncoupling of oxidative phosphorylation in the skeletal muscle of MDMA (40 mg/kg)-treated rats. In vivo, rats treated with MDMA had significantly elevated serum creatine kinase levels, a marker of rhabdomyolysis, 4 h post-MDMA treatment (955 +/- 132 IU/l) compared with saline-treated controls (373.2 +/- 59 IU/l). beta-ATP signal areas after MDMA treatment showed significant reductions (15%) from the baseline values with corresponding increases in inorganic phosphate (88% increases) and decreases in intracellular pH. Clark electrode experiments on isolated skeletal muscle mitochondria in vitro (1-5 mM MDMA) and ex vivo in MDMA-treated animals demonstrated no evidence of uncoupling of oxidative phosphorylation. In vitro experiments using L6 myotubules cocultured with primary hepatocytes demonstrated the presence of uncoupling protein-3 in the L6 myotubules, but no evidence of a direct effect of MDMA or its potential metabolites on cellular creatine kinase concentrations. These findings suggest that MDMA uncouples skeletal muscle mitochondria in vivo but that this uncoupling is the result of indirect mechanisms.

Most perfused tumor cell experiments are performed at 37 degrees C, the normal healthy body temperature. However, the temperature of subcutaneously implanted tumors in small animals is generally 29-33 degrees C when the rectal temperature of the animal is maintained at 37 degrees C. We have investigated the acute effects of increasing the temperature of perfused radiation-induced-fibrosarcoma (RIF-1) tumor cells from 33 to 37 degrees C (30 min) on intracellular sodium (Na(i)+) , intracellular pH (pH(i)), and bioenergetic status. Heating the cells by 4 degrees C produced a reversible increase in Na(i)+, slight acidification and no change in nucleotide triphosphate to inorganic phosphate ratio (NTP/P(i)) as measured by shift-reagent-aided (23)Na and (31)P NMR spectroscopy. In the presence of 3 microM 5-(N-ethyl-N-isopropyl) amiloride (EIPA), a potent and specific inhibitor of Na(+)/H(+) antiporter, the increase in Na(i)+ during the heating was completely abolished suggesting that the heat induced increase in Na(i)+ was caused by an increase in Na(+)/H(+) antiporter activity. However, the changes in pH(i) with the heating were identical with or without EIPA, indicating that pH(i) is controlled by other ion exchange mechanisms in addition to Na(+)/H(+) antiporter. NTP/P(i) was significantly higher in presence of EIPA for some time points during the heating suggesting that both NTP production and consumption rates may be altered during the heating. These results indicate that a slight increase in temperature from 33 to 37 degrees C induces significant changes in Na(+) physiology largely because of activation of Na(+)/H(+) antiporter but other ion exchange mechanisms are also involved in maintaining pH(i) in the RIF-1 tumor cells. Thus, care must be taken in choosing the temperature for perfused cell studies.