In vivo R1-enhancement mapping of canine myocardium using ceMRI with Gd(ABE-DTTA) in an acute ischemia-reperfusion model.

PURPOSE: To demonstrate the usefulness of normalized DeltaR1 (DeltaR1(n)) mapping in myocardial tissue following the administration of the contrast agent (CA) Gd(ABE-DTTA). MATERIALS AND METHODS: Ischemia-reperfusion experiments were carried out in 11 dogs. The method exploited the relatively long tissue lifetime of Gd(ABE-DTTA), and thus no fast R1 measurement technique was needed. Myocardial perfusion was determined with colored microspheres (MP). RESULTS: With varying extent of ischemia, impaired wall motion (WM) and lower DeltaR1(n) values were detected in the ischemic sectors, as opposed to the nonischemic sectors where normal WM and higher DeltaR1(n) were observed. Based on the DeltaR1(n), data from the myocardial perfusion assay and the DeltaR1(n) maps were compared in the ischemic sectors. A correlation analysis of these two parameters demonstrated a significant correlation (R = 0.694, P < 0.005), validating the DeltaR1(n)-mapping method for the quantitation of ischemia. Similarly, pairwise correlations were found for the MP, DeltaR1(n), and wall thickening (WT) values in the same areas. Based on the correlation between DeltaR1(n) and MP, DeltaR1(n) maps calculated with a pixel-by-pixel resolution can be converted to similarly high-resolution myocardial perfusion maps. CONCLUSION: These results suggest that the extent of the severity of ischemia can be quantitatively represented by DeltaR1(n) maps obtained in the presence of our CA.

The effects of the NMR shift-reagents Dy(PPP)2, Dy(TTHA) and Tm(DOTP) on developed pressure in isolated perfused rat hearts. The role of shift-reagent calcium complexes.

The 23Na NMR shift-reagent complexes (Dy(PPP)2, Dy(TTHA), and Tm(DOTP)) bind stoichiometric amounts of Ca2+. Thus, in perfused rat heart systems, a supplementation of Ca2+ is required to maintain the requisite extracellular free calcium concentration ([Ca(o)]f) and to approximate a physiological level of contractile function. The amount of reagent-bound Ca2+ in a heart perfusate that contains a shift-reagent depends on: (1) Ca2+ binding by excess ligand used during the preparation of the shift-reagent; and (2) the Ca2+ binding affinity of the shift-reagent. To address point 1), we introduced a 1H and 31P NMR spectroscopic titration method to quantify directly the concentration of the excess ligand. We also used this method to minimize the amount of excess ligand (L) and thus the amount of Ca*L complex. To address point (2), we determined the stepwise Kd (microm) values of the Ca complexes of the three shift-reagents.: Dy(PPP)2, Kd=0.09, Kd2=7.9; Dy(TTHA), Kd1=10.66, Kd2=10.12; and Tm(DOTP), K(d1)=0.502, Kd2=4.98. The Kd values of the Ca complexes of the phosphonate and triphosphate based shift-reagents, Tm(DOTP) and Dy(PPP)2, respectively, are lower than those of the polyaminocarboxylate-based Dy(TTHA), indicating stronger Ca binding affinities for the former two types of complexes. We have also shown a positive correlation between [Ca(o)]f and left ventricular developed pressure (LVDP) in perfused rat hearts. Dy(TTHA) has shown no effect on LVDP v[Ca(o)]f. The LVDP values in the presence of the phosphonate and triphosphate based shift-reagents, however, were significantly higher than expected from the [Ca(o)]f levels alone. Thus a positive inotropic effect, independent of [Ca(o)]f, is evident in the presence of Tm(DOTP) or Dy(PPP)2.

23Na NMR shift reagents enhance cardiac staircase effect in isolated perfused rat hearts.

The effects of the currently used (23)Na NMR shift reagents, dysprosium bis-triphosphate [Dy(PPP)(2)], dysprosium triethylenetriamine hexaacetate [Dy(TTHA)] and thulium 1,4,7, 10-tetraazacyclododecane-N,N',N",N"'-tetra(methylenephosphonate) [Tm(DOTP)] were studied in the rat heart cardiac staircase model. Rat hearts were perfused with low or normal extracellular free calcium ([Ca(o)](f)). At low [Ca(o)](f) (0.34 +/- 0.05 mM), hearts were perfused with Dy(PPP)(2) (group I), Dy(TTHA) (group II) or no shift reagent (group III), while at normal [Ca(o)](f) (1.25 +/- 0.15 mM), hearts were perfused with Tm(DOTP) (group IV), Dy(TTHA) (group V) or no shift reagent (group VI). Left ventricular developed pressure (LVDP) values in group I were significantly higher than in groups II and III (p < 0.01), while no significant differences were found between groups II and III. LVDP values in group IV were significantly higher than in groups V and VI (p < 0.05), while the LVDP values in groups V and VI were almost identical. Also, a positive correlation between pacing rate and intracellular sodium ([Na(i)]) was evident. The [Na(i)] values at high [Ca(o)](f) were significantly lower than at low [Ca(o)](f) at each pacing level (p <0.01), indicating a negative correlation between [Na(i)] and [Ca(o)](f). No statistical differences were found in [Na(i)] between groups I vs II and IV vs V, showing that determination of [Na(i)] is not affected by any of these shift reagents. Thus the different LVDP responses in groups I vs II and IV vs V were not mirrored in [Na(i)] changes. We hypothesize that a direct, sarcolemmal Ca-Dy(PPP)(2)-, or Ca-Tm(DOTP)-induced positive inotropic effect could be responsible for these Na(i)-independent LVDP increases in groups I and IV.

Synthesis and in vivo evaluation of new contrast agents for cardiac MRI.

Analogues 2-6 of N(3),N(6)-bis(2'-myristoyloxyethyl)-1, 8-dioxotriethylenetetraamine-N,N,N',N'-tetraacetic acid (BME-DTTA) (1), which like 1 are also based on the DTTA structure but contain shorter fatty acyl chains, were synthesized to improve the water solubility of the corresponding gadolinium complexes. The gadolinium complexes of 1 and 3-5 have very low solubility in water. Thus liposomal preparations are necessary for their in vivo MRI application. These liposomal preparations retain high in vitro relaxivities (27.1, 21.57, 20.32, 23.1 s(-1) mM(-1), respectively) and induce sustained MRI signal intensity enhancements (67.2, 38.4, 52.1, 41.7 in arbitrary units, respectively). The gadolinium complex of 2 is quite soluble in water. Its lifetime in the blood stream, however, is short. The gadolinium complex of analogue 6, N-(2-butyryloxyethyl)-N'-(2-ethyloxyethyl)-N,N'-bis[N' ',N' '-bis(carboxymethyl)acetamido]-1,2-ethanediamine (ABE-DTTA), has demonstrated its potential as a water-soluble, cardiac-specific, MRI contrast agent. It is completely soluble in water at a 25 mM concentration, allowing the preparation of an injectable dose. The in vitro relaxivity of the complex is 16.24 s(-1) mM(-1). The agent shows a specific accumulation in the heart tissue reaching its maximum within 15 min after administration, inducing a sustained MRI signal intensity enhancement of 43.6%. This enhancement lasts for at least 3 h, thus indicating a reasonably long lifetime of this contrast agent in the myocardium without deleterious effects on heart function parameters.

Lateralization of human temporal lobe epilepsy by 31P NMR spectroscopic imaging at 4.1 T.

OBJECTIVE: To compare the phosphorous metabolite ratios in the mesial temporal lobe of healthy volunteers (n = 20) with the corresponding ratios in patients with temporal lobe epilepsy (n = 30) using 31P NMR spectroscopic imaging and to lateralize the seizure focus in temporal lobe epilepsy patients using various phosphorous metabolite ratios-phosphocreatine to inorganic phosphate (PCr/Pi), PCr to adenosine triphosphate (PCr/gamma-ATP), and (gamma-ATP/Pi)--and to compare with clinical lateralization results. METHODS: All 31P NMR spectroscopic imaging studies were performed on a high-field, 4.1 T, whole-body NMR spectroscopic imaging system using a 31P/1H double-tuned volume coil. RESULTS: We found an average reduction of 15% in the PCr/Pi and gamma-ATP/Pi ratios compared with the corresponding ratios in healthy volunteers in the entire mesial temporal lobe, and more than a 30% reduction in these two ratios in the anterior region of the epileptogenic mesial temporal lobe. These ratios were also reduced significantly in the ipsilateral lobe when compared with their corresponding values in the contralateral lobe. In patients we lateralized the seizure focus, based on these 31P NMR data, and compared the results with the clinical lateralization. The lateralization based on either the PCr/Pi or the gamma-ATP/Pi ratio yielded a correspondence of 70 to 73% with the final clinical lateralization. In the subgroup of patients (n = 9) that needed intracranial EEG for the presurgical lateralization because of inconclusive results from the noninvasive methods, a 78% correspondence was found with the 31P NMR-based lateralization, whereas MRI provided a correspondence of only 33%, and scalp EEG provided a correspondence of only 56%. CONCLUSIONS: These results suggest the utility of adding the 31P NMR method to the group of noninvasive modalities used for presurgical decision making in temporal lobe epilepsy patients.

The modulation of pacing-induced changes in intracellular sodium levels by extracellular Ca2+ in isolated perfused rat hearts.

This study demonstrates the inverse relationship between extracellular free calcium ([Ca(o)]f) and intracellular sodium ([Na(i)]) in isolated perfused rat hearts and thus supports the role of [Na(i)] in the "calcium paradox". It also shows that the extent of the increase in [Na(i)] (delta[Na(i)]), and the extent of the decrease in left ventricular developed pressure (deltaLVDP) in isolated perfused rat hearts, induced by pacing, is modulated by [Ca(o)]f. At low (0.24 mM) as well as normal (1.15 mM) [Ca(o)]f, [Na(i)] increased with pacing, progressively and significantly (P<0.01 and P<0.05, respectively), reaching a maximum of 12.56 +/- 0.46 and 9.22 +/- 0.16 mM at 500 beats/min, respectively. At high [Ca(o)]f (2.2 mM), however, no pacing-induced increase in [Na(i)] was observed. Simultaneously, within the pacing range of 250-500 beats/min, the interval-force relationship was negative for all [Ca(o)]f. With decreasing [Ca(o)]f, a gradually increasing delta[Na(i)] was induced. We hypothesise that a [Ca(o)]f-dependent Na-Ca exchanger activity modulates Na+ uptake, and thus baseline [Na(i)]. During incremental pacing, the increase in pacing rate induces a [Ca(o)]f-dependent delta[Na(i)], which may interact further with the sarcolemmal Na-Ca exchanger activity. As a result, both baseline [Na(i)] and the pacing-induced, [Ca(o)]f-dependent delta[Na(i)] modulate the net Ca2+ uptake, and thus SR Ca, in a manner that results in a modulated left ventricular force development.

In vivo characterization of Gd(BME-DTTA), a myocardial MRI contrast agent: tissue distribution of its MRI intensity enhancement, and its effect on heart function.

We have determined an LD50 of 0.56 +/- 0.05 mmol/kg for liposomal Gd(BME-DTTA) in mice and also shown that liposomal Gd(BME-DTTA) has no deleterious effects on heart rate, blood pressure, left ventricular force and AV conductance in ferret hearts in vivo at the magnetic resonance imaging (MRI)-effective dose of 0.05 mmol/kg body weight. In MRI images, a 1H signal intensity enhancement is observed in the following organs in decreasing order of the effect: heart approximately spleen > kidney > liver. This enhancement is stable for over 3 h in all organs. The results of 1H MRI and electron micrographs indicate that the lipophilic fatty acyl groups in the ligand BME structure and the particle sizes of liposomal Gd(BME-DTTA) are two important factors for tissue specificity of liposomal Gd(BME-DTTA) in the intensity enhancement. In vitro relaxivity of a liposomal Gd(BME-DTTA) sample, stored at 4 degrees C, remained stable for over 4 months of observation, but a significant decrease in relaxivity was observed in a sample stored at room temperature, most likely reflecting some deterioration in liposome chemistry.

Is the intracellular pH different from normal in the epileptic focus of patients with temporal lobe epilepsy? A 31P NMR study.

We performed in vivo 31P NMR spectroscopic studies of human brain on a 4.1 T whole-body NMR system. Based on a control group of 20 healthy volunteers, the normal pHi was 7.05 (SD, 0.06; SEM, 0.01) in the left temporal lobe and 7.04 (SD, 0.04; SEM, 0.01) in the right temporal lobe. We also studied a patient group consisting of 13 individuals with unilateral temporal lobe epilepsy. The mean pHi was 7.02 (SD, 0.04; SEM, 0.01) in the ipsilateral lobe and 7.02 (SD, 0.05; SEM, 0.01) in the contralateral lobe. These results clearly show that no statistically significant difference in pHi is observed between the two lobes, either in normal controls or in patients. Also, no significant pHi difference exists between the control group and the patient group. Lateralization in each of the 13 patients with unilateral epilepsy, based on their individual pHi difference between the ipsilateral lobe and contralateral lobe (delta pHi), showed that three patients were nondiagnostic cases because their delta pHis were not significantly different from zero (< or = 0.02), five patients showed small delta pHis consistent with their clinical lateralization, whereas the remaining five patients showed delta pHi-based lateralization opposite to the clinical findings. These results seem to indicate an essentially random distribution around delta pHi = 0 within a very small experimental error of +/-0.02 pH units. pHi obtained from eight different areas in each of the 13 unilateral patients also did not show any significantly nonzero delta pHi values. These results led to the conclusion that even at the excellent spectral resolution and reproducibility of the 4.1 T machine (typical SD of 0.05 pH units), no significant pHi effect, induced by temporal lobe epilepsy, could be detected. Therefore, in this study, delta pHi does not appear to be a clinically useful tool for the lateralization of epileptic foci in patients with temporal lobe epilepsy.

In vivo MRI visualization of acute myocardial ischemia and reperfusion in ferrets by the persistent action of the contrast agent Gd (BME-DTTA).

BACKGROUND: Contrast agent-enhanced magnetic resonance imaging (MRI) has the potential to visualize myocardial ischemia. To date, however, no agent has been found that has a sustained effect that allows MRI detection for the entire duration of ischemia and reperfusion and thus is useful in conjunction with stress test MRI. In this article, we introduce the gadolinium complex of N3,N6-bis(2'-myrisotyloxyethyl)-1,8-dioxo-triethylene- tetraamine-N,N,N1,N1-tetraacetic acid [Gd(BME-DTTA)], an agent potentially useful for such a purpose. METHODS AND RESULTS: Four protocols were carried out. ECG-triggered, partially T1-weighted, spin-echo MRI was used in protocols A through C. In protocol A, in nonischemic ferrets, 50 mumol/kg Gd(BME-DTTA) induced a 70 +/- 5% intensity enhancement lasting 3 hours. In protocol B, the left anterior descending coronary artery was occluded, and a 99mTc-sestamibi-induced autoradiographic contrast verified (r = .87, P < .01) a Gd(BME-DTTA)-induced (n = 5) or Gd(DTPA)-induced (n = 4) MRI contrast. In the Gd(BME-DTTA) group a sustained contrast and in the Gd(DTPA) group a short-lived contrast were observed. In protocol C (n = 11), during ischemia, a 31 +/- 3.3% (P < .02) contrast was evident between the ischemic and nonischemic myocardial regions. Upon reperfusion, a contrast of 19 +/- 3% (P < .05) and 13 +/- 4.5% (P < .05) persisted for 5 and 15 minutes, respectively. Beyond 15 minutes, the contrast continued to diminish gradually. Nonradioactive microspheres verified (r = .87, P < .05) ischemia and reperfusion in this model. In protocol D (n = 4), blood delta R1 data showed that the blood pool retained Gd(BME-DTTA) for the entire time frame of the experiment at high enough concentration to provide an appropriate wash-in effect during the initial contrast enhancement and during reperfusion. CONCLUSIONS: This study demonstrates that Gd(BME-DTTA) induces a sustained MRI contrast between regions of normal versus ischemic myocardium, showing the potential of this agent for the diagnosis of ischemic heart disease in conjunction with stress tests.

Cardiac staircase and NMR-determined intracellular sodium in beating rat hearts.

Isolated, perfused rat hearts (30 degrees C, n = 13) were paced from 218 +/- 4 beats/min to 433 +/- 4 beats/min while systolic and diastolic pressure were recorded and intracellular Na+ concentration ([Na+]i) was monitored by 23Na nuclear magnetic resonance (NMR) spectroscopy. [Na+]i increased progressively with increasing stimulation frequency. In seven hearts (group I) an initial, progressive increase in systolic pressure was observed followed by a decrease in pressure with further increase in frequency. From the onset, a progressive decrease in systolic pressure was observed in group II (n = 6) in response to increased frequency. In group I an [Na+]i increase of up to 134 +/- 7% of control (P < 0.001) was observed, whereas in group II the gain in [Na+]i with increasing pacing rate was attenuated, reaching a maximum of 120 +/- 3% of control (P < 0.02). The differential pressure response between group I and group II hearts may reflect an enhanced sensitivity of rat hearts to the shortening of the restitution period of the sarcoplasmic reticulum, outweighing the positive inotropic effect induced by an increased [Na+]i. Only in rat hearts whose [Na+]i-induced increase in pressure outweights the restitution deficit would a complete positive inotropic effect be anticipated.

Gadolinium and dysprosium chelates of DTPA-amide-dextran: synthesis, 1H NMR relaxivity, and induced 23Na NMR shift.

In this study the conjugated macromolecular ligand, diethylenetriaminepentaacetic acid (DTPA)-amide-dextran, was synthesized by attaching DTPA to the dextran macromolecule (M(r) approximately 6000) by a covalent amide bond. Subsequently, DTPA-amide-dextran was complexed with either of the two lanthanide metal ions dysprosium (Dy) or gadolinium (Gd). The paramagnetic 23Na NMR shift induced by Dy(DTPA-amide-dextran) and the relaxivity (rho 1) induced by Gd(DTPA-amide-dextran) were characterized. Dy(DTPA-amide-dextran) induced a 25% larger 23Na NMR shift than that induced by Dy(DTPA). Neither the shift induced by Dy(DTPA-amide-dextran) nor the shift induced by Dy(DTPA) was affected by increasing levels of calcium ions in the solution. offDTPA-amide-dextran) exhibited an in vitro rho 1 of 8.4 (mM s)-1 at a 0.23 T magnetic field and 9.3 (mM s)-1 at a 0.47 T magnetic field, thus indicating a positive magnetic field dependence.

A selective inversion recovery method for the improvement of 23Na NMR spectral resolution in isolated perfused rat hearts.

Shift-reagent-aided 23Na NMR spectroscopy allows differentiation of the intracellular (Na(i)) and extracellular sodium (Na(o)) signals. The goal of the present study has been to develop a 23Na NMR spectroscopic method to minimize the intensity of the shift-reagent-shifted Na(o) signal and thus increase Na(i) resolution. This is achieved by a selective inversion recovery (SIR) method which enhances the resolution between the Na(i) and Na(o) peaks in shift-reagent-aided 23Na NMR spectroscopy. The application of SIR with Dy(TTHA), Tm(DOTP), or with low concentrations of Dy(PPP)2 results in both good spectral resolution and physiologically acceptable contractile function in the isolated, perfused rat heart model.

In vivo 31P nuclear magnetic resonance spectroscopy of human temporal lobe epilepsy.

We performed localized 31P nuclear magnetic resonance (NMR) 1H-image-guided in vivo spectroscopy to study regional high-energy phosphate levels in the brains of normal controls and in patients with intractable unilateral temporal lobe epilepsy. We did not observe differences in intracellular pH between controls and patients. The phosphocreatine/inorganic phosphate ratio was reduced by 50% in the epileptogenic temporal lobe compared with controls (p less than 0.005) and by 35% when compared with the unaffected contralateral temporal lobe (p less than 0.05). We did not observe differences in the ratio of phosphomonoesters to phosphodiesters between controls and patients. These findings suggest that in vivo 31P NMR spectroscopy yields a distinctive interictal metabolic profile in patients with intractable unilateral temporal lobe epilepsy and may permit noninvasive lateralizing evidence of the seizure focus.

Amiloride in ouabain-induced acidification, inotropy and arrhythmia: 23Na & 31P NMR in perfused hearts.

The increase in intracellular sodium (Nai), resulting from inhibition of the Na/K ATPase by cardiac glycosides, is known to increase calcium influx via Na(+)-Ca2+ exchange, and thereby increase contractility. This increase in intracellular Ca2+ has been related to the development of intracellular acidification and enhanced activity of the Na(+)-H+ exchanger as a measure by the cell to prevent further acidification. Thus, the efflux of the H+ ions results in an additional increase in Nai. This may subsequently lead to an increased rate of Ca2+ influx and therefore to the potentiation of the effects of cardiac glycosides. To assess the role of Na(+)-H+ exchange in the mechanism of ouabain action in the beating heart we used amiloride, a known inhibitor of Na(+)-H+ exchange. Isolated rat hearts were perfused with either ouabain (50 microM) alone (n = 8, Group I), amiloride (1.0 mM) + ouabain (50 microM) (n = 8, Group II), or amiloride (1.0 mM) alone as a control group (n = 4, Group III). 23Na and 31P NMR spectroscopy were used to assess the changes in Nai and intracellular pH (pHi), respectively, while simultaneous and continuous monitoring of left ventricular pressure was carried out. Perfusion with both ouabain alone (Group I) or ouabain + amiloride (Group II), resulted in a time dependent increase in Nai levels, reaching (within 25 mins) a maximum of 200 +/- 7% of control in Group I, and 170 +/- 10% of control in Group II. Concurrently, a mild but significant decrease in pHi was observed in both groups. This decrease, however, was significantly higher in Group II compared to Group I (0.34 pH units vs. 0.19 pH units, respectively; P less than 0.05), suggesting that inhibition of Na(+)-H+ exchange by amiloride limits the recovery from ouabain-induced intracellular acidification. While developed pressure gradually increased in Group I to a maximum of 268 +/- 52% of control, the addition of amiloride in Group II substantially reduced the positive inotropic effect. Ventricular fibrillation (VF) developed in three of the eight hearts in Group I within 10-13 mins after the addition of ouabain. Interestingly, the rate of Nai increase in hearts that sustained VF was significantly higher compared to those without VF (mean slope 10.1 +/- 2.11 vs. 3.9 +/- 1.0, respectively; P less than 0.0001). Ventricular fibrillation did not develop in Group II or III.(ABSTRACT TRUNCATED AT 400 WORDS)

The effect of postinfarction intramyocardial hemorrhage on transverse relaxation time.

1H NMR imaging has been used to define zones of myocardial infarction (MI), which appear as areas of relatively increased signal intensity (SI). However, zones of decreased SI have been observed within or around the areas of infarction in NMR images acquired at high magnetic fields. To determine the cause of these areas of reduced SI, ex vivo spin-echo 1H NMR imaging at 1.5 T was performed in eight dogs following 72 h of coronary artery occlusion. In all dogs, a zone of increased SI (122 +/- 7% compared to control myocardium; P less than 0.01) was observed in the territory of the occluded coronary artery. In seven of the dogs, additional zones were also seen, within or around the central zone of increased SI, which displayed SI that was reduced in comparison with the local enhanced intensity, but was similar to the intensity of normal myocardium (97 +/- 7% compared to control; P = NS). Gross inspection and histological assessment of sliced myocardium disclosed hemorrhage in these regions characterized by locally decreased NMR SI. Image-derived calculation of T2 in the various infarct regions revealed a significant shortening of T2 in the hemorrhagic infarct zones characterized by decreased SI, in comparison with the nonhemorrhagic infarct zones characterized by increased SI (59 +/- 7 ms vs 73 +/- 10 ms, P less than 0.05). No difference was found, however, between the observed T2's of hemorrhagic infarct and of control tissue (57 +/- 4 ms). Using a biexponential analysis of T2 from the hemorrhagic infarct zones, the intrinsic T2 of water protons affected by hemorrhage was determined to be 43 +/- 9 ms, significantly reduced in comparison with the values obtained with the standard monoexponential fit. The reduction in T2 in the hemorrhagic zone is consistent with the paramagnetic effects of deoxyhemoglobin associated with intramyocardial hemorrhage. Thus the apparent T2, measured in hemorrhagic infarct tissue, represents the result of an averaging effect of infarct and hemorrhage on T2 relaxation times. These observations improve our understanding of the changes in NMR SI within the infarcted regions, and may provide a noninvasive method for the detection and quantitative assessment of intramyocardial hemorrhage.

The longitudinal relaxation time (T1) of the intracellular 23Na NMR signal in the isolated perfused rat heart during hypoxia and reoxygenation.

Each of six perfused rat hearts was subjected to 30 min of hypoxia followed by 60 min of reoxygenation. Inversion-recovery data on the intracellular Na NMR signal, differentiated by a shift reagent, 6 mM Dy(PPP)2, were obtained every 5 min, and T1 values were calculated. The T1 of the intracellular Na signal did not show any significant change either during hypoxia or upon reoxygenation, although the level of Nai increased about 50%. Such an increase of total Nai is expected to reduce the observed relaxation rate by diluting the fraction of Nai ions that interact with intracellular polyelectrolytes. The observed constancy of T1 in our study is explained on the basis of the typical values of the dissociation constants of sodium ions, in aqueous solutions, in interaction with polyelectrolytes. Although the constancy of intracellular sodium T1 during hypoxia may preclude the utilization of T1 weighting for the monitoring of pathology, its determination could be important for setting optimal acquisition times in high time-resolution experiments.

Gadolinium complexes of [(myristoyloxy)propyl]diethylenetriaminetetraacetate: new lipophilic, fatty acyl conjugated NMR contrast agents.

New lipophilic contrast agents, 1-[3'-(myristoyloxy)propyl]diethylenetriamine-1,4,7,7-tetraacetic acid (1MP-DTTA), 4-[3'-(myristoyloxy)propyl]diethylenetriamine-1,1,7,7-tetraacetic acid (4MP-DTTA), and 4-[3'-(myristoyloxy)propyl]-2,6-dioxodiethylenetriamine-1,1, 7, 7-tetraacetic acid (4MPD-DTTA), were prepared from either diethylenetriamine or 3-amino-1-propanol (overall yield 16-23%). Liposome-incorporated Gd complexes of ligands 1MP-DTTA, 4MP-DTTA, and 4MPD-DTTA were prepared by mixing GdCl3 and the prepared vesicles consisting of ligand, egg lecithin, and cholesterol (molar ratio 1.1:5.1) followed by further sonication, and their in vitro relaxivities were determined. The relaxivities of these agents were higher than those of the Gd3+ aquoion, Gd(EDTA), or Gd(DTPA) at both 0.23 and 0.47 T. Gd(4MPD-DTTA) displayed the highest relaxivities (24.0 +/- 0.4 and 34.7 +/- 0.4, at 0.23 and 0.47 T, respectively) among these new Gd complexes. The relaxivities of these three agents increased from the lower to the higher magnetic field, indicating a positive field dependence. Stability constants (log K) of Gd(1MP-DTTA), Gd(4MP-DTTA), and Gd(4MPD-DTTA) were found to be 18.2 +/- 0.2, 18.4 +/- 0.2, and 15.7 +/- 0.8, respectively. A lower limit of 0.3 mmol/kg was found for LD50 for these three agents.

Fatty-acyl iminopolycarboxylates: lipophilic bifunctional contrast agents for NMR imaging.

New fatty-acyl contrast agents, N3-2'-myristoyloxyethyl-N6-2'-hydroxyethyl-1, 8-dioxotriethylenetetraamine-N,N,N',N'-tetraacetic acid (MHE-DTTA) and N3,N6-bis(2'-myristoyloxyethyl)-1,8-dioxo- triethylenetetraamine-N,N,N',N'-tetraacetic acid (BME-DTTA) were prepared by sequential alkylation, acylation, and catalytic hydrogenation from bis(hydroxyethyl)-ethylenediamine with satisfactory yields (overall 36-46%). The 1:1 gadolinium complexes of the ligands MHE-DTTA and BME-DTTA were incorporated into liposomes and their relaxivities in vitro were determined. The relaxivities of both agents were similar and were greater than those of Gd3+ aquoion, Gd(EDTA), and Gd(DTPA) at both 0.23 T and 0.47 T. The relaxivities of these two agents increased from the lower to the higher magnetic field, indicating a positive field dependence. This is advantageous because of the widespread use of high-field (B0 greater than 0.5 T) NMR imaging instruments. Stability constants (log K) of Gd(MHE-DTTA) and Gd(BME-DTTA) were found to be 15.27 +/- 2.21 and 16.78 +/- 0.36, respectively. LD50 of both compounds was greater than 0.2 mmol/kg. These stabilities and lower limits of LD50 indicate the possible in vivo application of these agents.

Improvement of spectral resolution in shift-reagent-aided 23Na NMR spectroscopy in the isolated perfused rat heart system.

The level of intracellular sodium (Nai) is maintained at approximately 14 mM in healthy myocytes. When myocytes are damaged, Nai increases and therefore the level of Nai may be a means of evaluating myocardial cell integrity. A particularly useful method to monitor Nai levels is 23Na NMR spectroscopy. However, because of the isochronous nature of the extracellular sodium (Nao) and Nai NMR signals, paramagnetic lanthanide shift reagents (LSR), such as dysprosium triphosphate, Dy(PPP)7-(2), have been used to shift the Nao signal. This reveals the unshifted Nai signal and allows the NMR monitoring of Nai in isolated perfused hearts and other systems. A major shortcoming of this method (the "shift-only" method) is in the need to minimize the Nao signal by not submerging the perfused hearts in Na(+)-containing buffer. An equally undesirable alternative is the utilization of relatively high concentrations of LSR to shift a large Nao signal sufficiently to enable reasonable resolution and quantitation of Nai. We present here a method, the "shift-relaxation" method, which is a combination of using a mixture of Dy(PPP)7-(2), a shift reagent, and gadolinium triphosphate, Gd(PPP)7-(2), a relaxation agent, with data acquisition using an inversion-recovery (IR) pulse sequence. This combination allows differentiation between Nao and Nai by the difference in their respective T1 values in addition to the shift between them. With this technique we can selectively minimize the extracellular signal and therefore minimize the need for a large Dy-induced shift, as well as allow data acquisition on a heart submerged in Na(+)-containing perfusate. The resulting improved discrimination between Nai and Nao at relatively low levels of LSR should be helpful for ultimate in vivo applications and potential clinical applications, where a lower dose of LSR also means a decreased possibility of physiologically deleterious effects. Also included in this paper is a method for the quick determination of an accurate 180 degrees pulse which is required for the optimization of the IR method.

Detection of intramyocardial hemorrhage using high-field proton (1H) nuclear magnetic resonance imaging.

Proton (1H) nuclear magnetic resonance (NMR) imaging has been used to define zones of myocardial infarction (MI), which appear as areas of relatively increased signal intensity (SI). However, zones of decreased SI have been observed within the areas of infarction and have been postulated to result from intramyocardial hemorrhage. To explore this phenomenon further, ex vivo spin-echo 1H NMR imaging at 1.5 Tesla was performed in 17 dogs after 24 hr (n = 9) and after 72 hr (n = 8) of coronary artery occlusion. In all dogs, a zone of increased SI (118 +/- 9% compared with normal myocardium) was observed in the distribution of the occluded coronary artery. In 12 of the 17 dogs, zones of decreased SI (92 +/- 8% compared with normal) were seen within or around the central zone of increased SI. Gross inspection and histological assessment of sliced myocardium usually disclosed hemorrhage in the regions of decreased SI. In three of the five dogs with no apparent zones of decreased SI on NMR, the infarct was small, and only minor hemorrhage was observed by gross inspection, whereas in the remaining two dogs no hemorrhage was seen. Myocardial flow in the hemorrhagic regions was significantly higher than in the necrotic core (59 +/- 29% vs. 31 +/- 24% compared with control, P less than 0.05). Image-derived calculation of T2 relaxation times in the different infarcted regions revealed a significant shortening of T2 in the infarcted hemorrhagic zones with decreased SI compared with the infarct zones with increased SI (49 +/- 8 msec vs. 66 +/- 8 msec, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)