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Proton magnetic resonance imaging (MRI) and histological methods were used to evaluate photodynamic therapy (PDT)-induced hemorrhagic necrosis in the murine M1 tumor within 72 h of treatment of male DBA/2 mice. The effects of three photosensitizing drugs were investigated: Photofrin (n = 4), Zn (II) phthalocyanine (n = 7) and benzoporphyrin derivative monoacid ring A (n = 11). As noted in previous studies of PDT using MRI, MRI makes possible serial, noninvasive, in vivo observation of tissue response to PDT. Our serial study of MRI and histological data confirms that tumors responded in the same way to PDT treatment using the three photosensitizing drugs: vascular damage followed by hemorrhagic necrosis. Most importantly and unlike previous MRI studies of PDT, we used a very high field magnet that enhanced the effect of magnetic susceptibility on image signal when blood is processed by the body after PDT-induced hemorrhagic necrosis. This last finding demonstrates the utility of high field magnets and the importance of localized, serial experiments in future magnetic resonance studies of PDT.

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To evaluate the effect of hypothermic circulatory arrest on brain metabolism, we used 31P-magnetic resonance spectroscopy to monitor brain metabolites in pigs during 2 hours of ischemia and 1 hour of reperfusion. Twenty-eight pigs were divided into five groups. Anesthesia (n = 5) and hypothermic cardiopulmonary bypass groups (n = 5) served as controls. In the circulatory arrest (n = 6), antegrade perfusion (n = 6), and retrograde (n = 6) brain perfusion groups, the bypass flow rate was 60 to 100 ml.kg-1.min-1. In the antegrade group, the brain was perfused via the carotid arteries at a blood flow rate of 180 to 200 ml.min-1 during circulatory arrest at 15 degrees C. In the retrograde group, the brain was perfused through the superior vena cava at a flow rate of 300 to 500 ml.min-1 during circulatory arrest at 15 degrees C. The intracellular pH was 7.1 +/- 0.1 and 7.3 +/- 0.1 in the anesthesia and hypothermic cardiopulmonary bypass groups, respectively. In the circulatory arrest group, the intracellular pH decreased to 6.2 +/- 0.1 and did not recover to its initial value (7.0 +/- 0.1) during reperfusion (p < 0.05 compared with the value obtained from the control groups at the corresponding time). Inorganic phosphate did not return to its initial level during reperfusion. In three animals in this group, levels of high-energy phosphates, adenosine triphosphate and phosphocreatine, recovered partially but did not reach the levels observed before arrest. In the group receiving antegrade perfusion, cerebral metabolites and intracellular pH were unchanged throughout the protocol. During circulatory arrest in the retrograde perfusion group the intracellular pH decreased to 6.4 +/- 0.1 and recovered fully during reperfusion (7.1 +/- 0.1). High-energy phosphates also returned to their initial levels during reperfusion. These studies show that deep hypothermic circulatory arrest with antegrade brain perfusion provides the best brain protection of the options investigated.

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Glycolipids constitute an important class of biomolecules that are involved in biomolecular recognition. The importance of carbohydrate head group conformation in such processes is well recognized. Glycolipids typically occur as minor components of the complex heterogeneous matrix of a biological membrane. As a result, the membrane surface may not only influence head group conformation but also serves as a spatial frame in which the glycolipid is oriented and recognized. In this study, conformational energy calculations have been used to assess the conformational space available to the glucose head group of 1,2-di-O-tetradecyl-3-O-(beta-D-glucopyranosyl)-sn-glycerol (beta-DTGL) in a liquid-crystalline membrane matrix. 2H NMR quadrupolar splittings are calculated and compared with those observed experimentally. This study demonstrates the importance of including surface interactions when considering the conformational space accessible to cell surface carbohydrates. The empirical approach taken here provides considerable insight at the molecular level, and offers the possibility of exploring even more complex systems.

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It has been shown previously that two types of motion are adequate to describe the partially relaxed 2H NMR line shapes (inversion recovery experiment) for the backbone portion of the glycolipid 1,2-di-O-tetradecyl-3-O-(beta-D-glucopyranosyl)-sn-glycerol (beta-DTGL) in the highly ordered gel phase (Auger, M.A., D. Carrier, I.C.P. Smith, and H. C. Jarrell. 1990. J. Am. Chem. Soc. 112:1373-1381). This study extends the latter investigation to the more fluid liquid-crystalline phase, where more complex motions are anticipated. Analyses of the powder line shapes and oriented sample relaxation data for both the glycerol backbone and head group regions of this lipid have been performed. The dynamics of glycerol at the C3 position in the gel state have been described by large angle jumps about the C2-C3 bond with a correlation time in the fast-limit motional regime (omega o tau c much less than 1) and site populations 0.46, 0.34, and 0.20. The present data show that in the liquid-crystalline phase the internal jump rate is maintained, and two additional motions are necessary to describe the dependence of the relaxation rate on the orientation of the director with respect to the magnetic field direction. These are rotation about the molecular long axis with a correlation time in the slow-limit motional regime very near to the T1 minimum (omega o tau c approximately 0.65), and molecular fluctuations about the order director (modeled by a Maier-Saupe restoration potential). This treatment was also extended to the glucose head group where additional segmental motion about the glycosidic bond has been reported previously. While the two motions dominating relaxation at the glycerol C3 segment reproduce the general relaxation features of the glucose head group, the results suggest that additional motion about the glycosidic linkage must be present. This study is a stringent test of the motional model chosen earlier because relaxation data were obtained at two 2H NMR frequencies using two relaxation experiments (T1Z and T1Q) and two types of sample preparation (oriented and dispersed multibilayers). The results strongly uphold the choice of model and indicate the utility of both oriented samples and the T1Q experiment.

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