Phospholipid bicelles with positive anisotropy of the magnetic susceptibility.
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Magnetic Resonance Spectroscopy↗
Biomedical subjects
B M Fung
Publications and source records attributed to B M Fung.
Indirect NMR detection in solids with multiple cross-polarization periods.
The use of indirect detection for signal enhancement in solids is much less common than in liquids, but has attracted renewed interest recently. In this work we describe an indirect detection scheme that offers a large signal enhancement for rare spins in solids. The method uses multiple periods of cross polarization, each followed by an evolution period. The latter is increased stepwise in a pseudo 2D experiment, in which the signal of the rare spin is detected as modulation of the abundant spin. As an illustration of this method, the natural abundance deuterium NMR spectrum of a static powder sample of 1,2,4,5-tetramethylbenzene is presented.
Deuterium↗
An improved broadband decoupling sequence for liquid crystals and solids.
Recently we developed an efficient broadband decoupling sequence called SPARC-16 for liquid crystals ¿J. Magn. Reson. 130, 317 (1998). The sequence is based upon a 16-step phase cycling of the 2-step TPPM decoupling method for solids ¿J. Chem. Phys. 103, 6951 (1995). Since then, we have found that a stepwise variation of the phase angle in the TPPM sequence offers even better results. The application of this new method to a liquid crystalline compound, 4-n-pentyl-4'-cyanobiphenyl, and a solid, L-tyrosine hydrochloride, is reported. The reason for the improvement is explained by an analysis of the problem in the rotating frame.
Crystallization↗
An efficient broadband decoupling sequence for liquid crystals.
A new and efficient broadband decoupling sequence is presented. It is based on the phase cycling of two pulses with small phase angles. The pulse width is 180 degrees +/- 30 degrees, and the phase angle is +/-10 degrees-12 degrees. The sequence contains 16 elements, and is called SPARC-16 as an abbreviation for small phase angle rapid cycling. The application of this sequence to a liquid crystalline compound, 4-n-pentyl-4'-cyanobiphenyl (5CB), is reported. The signal-to-noise ratios and the linewidths of the 13C peaks of 5CB with SPARC-16 decoupling and with other decoupling methods are compared. The results show that the broadband decoupling efficiency of SPARC-16 is considerably better than those of other methods.
Crystallization↗
Dynamic evaluation of clotting phenomena in vitro and perfluorochemical oxygen transport across a membrane-bound thrombus model.
Citrated platelet-rich plasma was used to occlude 3-microns and 10-microns poresize Nuclepore membranes after recalcification as a thrombus model. Morphologic studies, using both light microscopy and scanning electron microscopy, indicated that over 90 percent of the number of pores available for filtration in hydrophilic and hydrophobic membranes were occluded either partially or completely. Results of transient and steady state pressure drop measurements supported the morphologic studies. It was found that the percentage of oxygen transported across the occluded membranes was greater for filtration of red blood cell suspensions diluted with a perfluorochemical emulsion than that of those diluted with Ringers. The findings in this study suggested that perfluorochemical emulsions might transport oxygen across a thrombus to maintain tissue viability during acute ischemic events.
Adult↗
Thrombolysis using liposomal-encapsulated streptokinase: an in vitro study.
The clot-lysing ability of streptokinase (SK) was examined using membrane-bound thrombi. Encapsulation of SK in large unilamellar phospholipid vesicles (liposomes) resulted in entrapping approximately 30% of its original activity. Measurements of streptokinase activity for liposomal-encapsulated streptokinase (LESK) indicated little loss of activity or leakage in Tris-buffered saline over a 24-hr period at temperatures of 4 and 23 degrees C. However, incubation of free SK and LESK in platelet-poor plasma (PPP) at 37 degrees C resulted in a decrease of SK activity. The retention of SK activity in LESK was considerably higher than that of unentrapped SK. Clot-dissolving time (CDT) was measured by monitoring the pressure drop during slow filtration in plasma through membrane-bound thrombi. The results indicated that both LESK and free SK were able to activate the fibrinolytic system. Without prior incubation in PPP at 37 degrees C, the CDT of a SK and PPP mixture (SK/PPP) was 10.7 +/- 1.9 min (n = 12), while that of a LESK and PPP mixture (LESK/PPP) was 12.4 +/- 1.7 min (n = 12). The CDT-detected clot-lysing abilities of both SK and LESK were diminished by incubation in PPP, but to different extents. After 15- and 30-min incubations, the CDT of SK/PPP increased significantly to 15.5 +/- 1.5 and 24.1 +/- 2.4 min (n = 5, P less than 0.05), respectively. In contrast, the CDT of LESK/PPP increased to 13.3 +/- 0.8 min (n = 5) after 15 min of incubation and to 16.0 +/- 1.1 min (n = 5, P less than 0.05) after a 30-min incubation. These results suggest that entrapment of SK in liposomes preserves the thrombolytic potential of the plasminogen activator by limiting its exposure to the components of the plasma.
Adult↗
Interaction between erythrocytes and a perfluorochemical blood substitute.
Perfluorochemicals (PFCs) are being widely developed for use as erythrocyte substitutes to carry oxygen to tissue in acute crises. Oxypherol, a commercially available PFC preparation commonly used for animal tests, reduces erythrocyte deformability in the presence of plasma. This undesirable effect further complicates oxygen delivery by erythrocytes. Our experiments indicate that one or more plasma proteins must be present to observe Oxypherol-induced reduction in erythrocyte deformability, but the latter is not protein specific. Neither platelet activation nor enhanced protein adsorption could account for the mechanism of altered erythrocyte flexibility. Using fluorine-19 NMR, it was found that a small amount of Oxypherol droplets is adsorbed on the surface of the erythrocytes. The amount of adsorbed droplets increases when Oxypherol is incubated with the erythrocytes in the presence of plasma, in parallel with a decrease in erythrocyte deformability. Therefore, it seems likely that the loss of deformability of the erythrocytes is caused by the adsorption of small Oxypherol droplets on the erythrocytes.
Adsorption↗
Perfluorochemical emulsions with fluorinated surfactants and anticancer drugs.
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Adamantane↗
Nuclear magnetic resonance study of water interactions with proteins, biomolecules, membranes, and tissues.
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Animals↗
Plasma-mediated alterations of erythrocyte deformability by perfluorochemical blood substitutes.
The effect of perfluorochemical blood substitutes (eg, Oxypherol or Fluosol-DA) on red cell deformability was investigated because these emulsions are in direct contact with red cells when they are used as temporary circulatory aids. Erythrocyte deformability was assessed by a constant volumetric flow rate filtration method. The results of in vitro incubation experiments indicate that perfluorotributylamine causes the deformability of human red cells to decrease significantly in the presence of plasma. However, there is no obvious loss in the deformability when washed cells are used. Neither mean cell volume nor white cells appear to be responsible for the observed effects of perfluorotributylamine. Perfluorodecalin and perfluorotripropylamine, two perfluorochemical compounds that are widely applied clinically, do not induce significant changes in red cell deformability with or without plasma. These results indicate the need for in vitro testing in the development of perfluorochemicals as blood substitutes.
Blood Substitutes↗
The effect of ATP concentration on the rate of actin polymerization.
It was found that the rate of polymerization of G-actin increased with the decrease of ATP concentration. When excess ATP was replaced by chloride through anion-exchange treatment, the extent of actin polymerization did not change provided that the ionic strength was raised immediately after the treatment. In the meantime, the rate of actin polymerization was greatly enhanced after the removal of excess ATP. The rate enhancement was much less when both excess Ca2+ and excess ATP were removed. G-actin with excess ATP replaced by chloride had larger light scattering and showed a "catalytic" effect on the polymerization of normal G-actin. The inhibition of actin polymerization by cytochalasin B in 100 mM KCl was much more obvious for G-actin with excess ATP removed than for normal G-actin. It is suggested that the reduction of excess ATP concentration in a G-actin solution increases the binding of weak-affinity Ca2+ and promotes the formation of oligomeric actin (actin nuclei).
Actins↗
Nuclear magnetic resonance transverse relaxation in muscle water.
The origin of the nonexponentiality of proton spin echoes of skeletal muscle has been carefully examined. It is shown that the slowly decaying part of the proton spin echoes is not due to extracellular water. First, for muscle from mice with in vivo deuteration, the deuteron spin echoes were also nonexponential, but the slowly decaying part had a larger weighing factor. Second, for glycerinated muscle in which cell membranes were disrupted, the proton spin echoes were similar to those in intact muscle. Third, the nonexponentiality of the proton spin echoes in intact muscle increased when postmortem rigor set in. Finally, when the lifetimes of extracellular water and intracellular water were taken into account in the exchange, it was found that the two types of water would not give two resolvable exponentials with the observed decay constants. It is suggested that the unusually short T2's and the nonexponential character of the spin echoes of proton and deuteron in muscle water are mainly due to hydrogen exchange between water and functional groups in the protein filaments. These groups have large dipolar or quadrupolar splittings, and undergo hydrogen exchange with water at intermediate rates. The exchange processes and their effects on the spin echoes are pH-dependent. The dependence of transverse relaxation of pH was observed in glycerinated rabbit psoas muscle fibers.
Animals↗
Modification of cell membrane lipids in Micrococcus lysodeikticus induced by pantoyl lactone.
Growth of Microccoccus lysodeikticus in the presence of pantoyl lactone brings about both qualitative and quantitative changes in cell membrane lipids. Significant amounts of the two major phospholipids (phosphatidylglycerol and diphosphatidylglycerol) are converted to lyso forms; the largest conversion occurs in the phosphatidylglycerol. In addition, amounts of several phospholipid fatty acids are changed. Physical alteration of the call membrane can be demonstrated using differential scanning calorimetry. Although growth and transport are significantly inhibited when pantoyl lactone is present, cells possessing altered call membrane phospholipds and phospholipid fatty acids, brought about by growth in the presence of pantoyl lactone, transport D-alanine, L-glutamic and L-aspartic acid normally when washed free of the pantoyl lactone.
4-Butyrolactone↗
Varied magnetic field, multiple-pulse, and magic-angle spinning proton nuclear magnetic resonance study of muscle water.
The nuclear magnetic resonance linewidth of 1H in water of frog muscle was studied as a function of magnetic field strength and angle of orientation. The results suggest that the observed spectra are dominated by demagnetization field anisotropy and dispersion, but a small static dipolar interaction of the order of a few hertz man be present. Data from line-narrowing, multiple-pulse experiments also indicate the presence of a small dipolar broadening.
Animals↗
Study of spin-lattice and spin-spin relaxation times of 1H, 2H, and 17O in muscular water.
Spin-lattice (T1) and spin-spin (T2) relaxation times of proton, deuteron, and oxygen-17 in muscle water have been measured at 9.21 MHz in the temperature range of 0 degree--40 degrees C. The values of the apparent activation energy for the three nuclei are (in kJ . mol-1) 9.1, 19, and 18 for 1/T1, and -1.3, 4.2, and 14 for 1/T2, respectively. The relatively small values for T2 for 1H and 2H and their low apparent activation energies are attributed to hydrogen exchange between water and proteins; this exchange does not affect the 17O relaxation. Quantitative calculations on deuteron T1 and oxygen-17 T1 and T2 have been made. The effect of surface-induced anisotropy on a minor fraction of water molecules is considered in some detail, and a new expression for its spectral density similar to that of liquid crystalline systems is applied in the calculation. It is suggested that water on the surfaces of macromolecules has a rotational correlation time of tau c approximately 1 x 10(-9) S, with a time constant of tau x approximately 3 x 10(-7) S, which is characteristic of the relaxation of the local structure.
Animals↗
Correlation of relaxation time with water content in muscle and brain tissues.
Proton T1 of water in muscle and brain tissues was studied as a function of water content at 25 degrees C and 5, 30, and 100 MHz. Muscle samples were bathed in modified Krebs solutions that have the same ionic strength but different osmolarity in order to change the water content. At all three frequencies their 1/T1 extrapolated to the same value of 0.6 S-1 for zero dry weight, indicating that T1 for the majority of water molecules in normal muscle and brain is frequency-independent. Dehydrated muscle, homogenized brain tissue suspended in modified Krebs solution and dehydrated brain tissue provide samples with wide ranges of water content. Their T1 values at all three frequencies can be well accounted for by a simple equation derived from the two-state model.
Animals↗
Carbon-13 and proton magnetic resonance of mouse muscle.
It is shown that roughly 4 mmol carbon atoms/g mouse muscle can give rise to a "high resolution" 13C NMR spectrum. From the 13C spectrum, it is estimated that the protons from mobile organic molecules or molecular segments amount to 6-8%of total nonrigid protons (organic plus water) in muscle. Their spin-spin relaxation times (T2) are of the order of 0.4-2 ms. At 37 degrees C, the proton spin-echo decay of mouse muscle changes rapidly with time after death, while that of mouse brain does not.
Animals↗
Proton and deuteron relaxation of muscle water over wide ranges of resonance frequencies.
The spin-lattice relaxation time (T1) of water protons in mouse muscle was studied from 10(4) to 10(8) Hz at several temperatures, and the deuteron T1 of muscle water was studied from 2.0 X 10(3) to 1.54 X 10(7) Hz at several temperatures. Proton T1's of muscle and brain water with different D2O contents were measured at 25 degrees C and 35 MHz. From the results of variable frequency and temperature measurements and the data of isotope substitution, it is concluded that the major relaxation mechanism for the protons in muscle water is the intermolecular dipolar interaction between the protons of the macromolecules and the protons of the water molecules in the hydration layer. It is also suggested that the relaxation of deuterons can be accounted for a very small fraction of water molecules directly hydrogen-bonded to the macromolecules.
Animals↗