Determination of water compartments in rat myocardium using combined D-T1 and T1-T2 experiments.

We have used combined D-T1 and T1-T2 correlation experiments to explore water compartments in rat heart tissue (myocardium). The results show that two main compartments can be identified, which we assign to extracellular (ec) and intracellular (ic) water. The exchange rate of water across the cell membrane was found to be on the order of 0.1 Hz. In addition, the T1-T2 correlation measurements indicate that the ic compartment contain two T2 populations.

Manganese-calcium interactions with contrast media for cardiac magnetic resonance imaging: a study of manganese chloride supplemented with calcium gluconate in isolated Guinea pig hearts.

OBJECTIVES: Manganese ions (Mn) enter cardiomyocytes via calcium (Ca) channels and enhance relaxation intracellularly. To prevent negative inotropy, new Mn-releasing contrast agents have been supplemented with high Ca. The study aim was to investigate how this affects cardiac function and magnetic resonance efficacy. MATERIALS AND METHODS: MnCl2 based contrast agents, manganese and manganese-calcium (Ca:Mn 10:1), were infused during 4 repeated washin-washout sequences in perfused guinea pig hearts. [Mn] were 10, 50, 100 and 500 microM. RESULTS: During washin, manganese depressed left ventricular developed pressure (LVDP) by 4, 9, 17, and 53% whereas manganese-calcium increased LVDP by 13, 18, 25, and 56%. After experiments, tissue Mn contents (nmol/g dry wt) were control <40, manganese 3720, and manganese-calcium 1620. T1 was reduced by 85-92% in Mn-enriched hearts. CONCLUSIONS: High Ca supplements to Mn-releasing contrast agents may be counterproductive by inducing a strong positive inotropic response and by reducing the magnetic resonance efficacy.

Correlations between diffusion, internal magnetic field gradients, and transverse relaxation in porous systems containing oil and water.

The main focus in this study is to investigate the correlations between internal magnetic field gradients (G0) and transverse relaxation times in liquid-saturated packings of glass beads of different wettabilities. We show how these correlations can be expressed as two-dimensional (2D) diagrams of distribution functions between internal magnetic field gradients and T2 values. In the case where it is difficult to distinguish the signals from oil and water, we separate them based on their difference in diffusivity. In addition to using such diffusion weighting in the G0-T2 diagrams, we also show results from experiments where the direct correlation between diffusion and T2 (D-T2) is determined. The overall results show that the wettability of the glass beads has a strong influence on the appearance of these diagrams, in particular on the location of the fast diffusing water molecules. However, due to their lower diffusivity, the transverse magnetization of the oil molecules is not so greatly influenced by either the presence of the glass beads or their wettability properties. Thus, the wettability properties of a liquid-filled porous material can be determined from the location of the water signal in such 2D diagrams. In particular, we show that this is the case not only for D-T2 diagrams, but also for G0-T2 diagrams.

Intracellular manganese ions provide strong T1 relaxation in rat myocardium.

The efficacy of manganese ions (Mn2+) as intracellular (ic) contrast agents was assessed in rat myocardium. T1 and T2 and Mn content were measured in ventricular tissue excised from isolated perfused hearts in which a 5-min wash-in with 0, 30, 100, 300, or 1000 microM of Mn dipyridoxyl diphosphate (MnDPDP) was followed by a 15-min wash-out to remove extracellular (ec) Mn2+. An inversion recovery (IR) analysis at 20 MHz revealed two T1 components: an ic and short T1-1 (650-251 ms), and an ec and longer T1-2 (2712-1042 ms). Intensities were about 68% and 32%, respectively. Tissue Mn content correlated particularly well with ic R1-1. A two-site water-exchange analysis of T1 data documented slow water exchange with ic and ec lifetimes of 11.3 s and 7.5 s, respectively, and no differences between apparent and intrinsic relaxation parameters. Ic relaxivity induced by Mn2+ ions in ic water was as high as 56 (s mM)(-1), about 8 times and 36 times higher than with Mn2+ aqua ions and MnDPDP, respectively, in vitro. This value is as high as any reported to date for any synthetic protein-bound metal chelate. The increased rotational correlation time (tauR) between proton and electron (Mn2+) spins, and maintained inner-sphere water access, might make ic Mn2+ ions and Mn2+ -ion-releasing contrast media surprisingly effective for T1-weighted imaging.

Manganese ions as intracellular contrast agents: proton relaxation and calcium interactions in rat myocardium.

Paramagnetic manganese (Mn) ions (Mn(2+)) are taken up into cardiomyocytes where they are retained for hours. Mn content and relaxation parameters, T(1) and T(2), were measured in right plus left ventricular myocardium excised from isolated perfused rat hearts. In the experiments 5 min wash-in of MnCl(2) were followed by 15 min wash-out to remove extracellular (ec) Mn(2+) MnCl(2), 25 and 100 micro M, elevated tissue Mn content to six and 12 times the level of control (0 micro M MnCl(2)). Variations in perfusate calcium (Ca(2+)) during wash-in of MnCl(2) and experiments including nifedipine showed that myocardial slow Ca(2+) channels are the main pathway for Mn(2+) uptake and that Mn(2+) acts as a pure Ca(2+) competitor and a preferred substrate for slow Ca(2+) channel entry. Inversion recovery analysis at 20 MHz revealed two components for longitudinal relaxation: a short T(1 - 1) and a longer T(1 - 2). Approximate values for control and Mn-treated hearts were in the range 600-125 ms for T(1 - 1) and 2200-750 ms for T(1 - 2). The population fractions were about 59 and 41% for the short and the long component, respectively. The intracellular (ic) R(1 - 1) and R(2 - 1) correlated best with tissue Mn content. Applying two-site exchange analyses on the obtained T(1) data yielded results in parallel to, but also differing from, results reported with an ec contrast agent. The calculated lifetime of ic water (tau(ic)) of about 10 s is compatible with a slow water exchange in the present excised cardiac tissue. The longitudinal relaxivity of Mn ions in ic water [60 (s mM)(-1)] was about one order of magnitude higher than that of MnCl(2) in water in vitro [6.9 (s mM)(-1)], indicating that ic Mn-protein binding is an important potentiating factor in relaxation enhancement.

A putative DNA-binding domain in the NUCKS protein.

We have studied the DNA-binding properties of a NUCKS-derived, synthetic peptide containing an extended GRP motif. This peptide binds to random-sequence DNA, but did not bind preferentially to poly(dA-dT). A synthetic peptide with the same amino acid composition but with a random sequence did not bind to DNA, suggesting that the structure of the DNA-binding domain plays a pivotal role in the interaction with DNA. NMR and graphic modeling were employed to investigate the structure of the synthetic peptide. It was shown that the DNA-binding peptide constituted an alpha helix in phosphate buffer at pH 5.5. Docking results indicated an almost perfect fit for this small, helical peptide into the major groove of DNA with the possibility of four basic residues interacting with the phosphate backbone of DNA. One consensus site for phosphorylation by Cdk1 is located in the N-terminal end of the DNA-binding peptide. Upon phosphorylation of this site, the binding to DNA was completely prohibited. Immunofluorescence experiments showed that NUCKS was located in the nuclei in proliferating cells in interphase of the cell cycle, but was distributed throughout the cytoplasm in mitotic cells.