Enzymatic activity monitoring through dynamic nuclear polarization in Earth magnetic field.

Cost-effective and portable MRI systems operating at Earth-field would be helpful in poorly accessible areas or in developing nations. Furthermore Earth-field MRI can provide new contrasts opening the way to the observation of pathologies at the biochemical level. However low-field MRI suffers from a dramatic lack in detection sensitivity even worsened for molecular imaging purposes where biochemical specificity requires detection of dilute compounds. In a preliminary spectroscopic approach, it is proposed here to detect protease-driven hydrolysis of a nitroxide probe thanks to electron-nucleus Overhauser enhancement in a home-made double resonance system in Earth-field. The combination of the Overhauser effect and the specific enzymatic modification of the probe provides a smart contrast reporting the enzymatic activity. The nitroxide probe is a six-line nitroxide which lines are shifted according to its substrate/product state, which requires quantum mechanical calculations to predict EPR line frequencies and Overhauser enhancements at Earth field. The NMR system is equipped with a 13-mT prepolarization coil, a 153-MHz EPR coil and a 2-kHz NMR coil. Either prepolarized NMR or DNP-NMR without prepolarization provide NMR spectra within 3 min. The frequency dependence of Overhauser enhancement was in agreement with theoretical calculations. Protease-mediated catalysis of the nitroxide probe could only be measured through the Overhauser effect with 5 min time resolution. Future developments shall open the way for the design of new low-field DNP-MRI systems.

Muscle phosphocreatine post-exercise recovery rate is related to functional evaluation in hospitalized and community-living older people.

OBJECTIVE: to explore muscle mitochondria function with respect to age, functional status and nutrition in community-living and recovering hospitalized older subjects. MEASUREMENTS: subjects were assessed for nutrition, hand-grip strength, 10-meter gait time, a modified timed get-up-and-go test and activities of daily living score (ADL). 31P magnetic resonance spectroscopy (31P MRS) was used to assess the initial rate of post-exercise phosphocreatine recovery (ViPCr) for mitochondrial function evaluation in 25 hospitalized older subjects (86.1 + 5.3 y) and in 25 community-living younger ones (74.5 + 6.2 y). RESULTS: in multiple linear regression, longer time on the get-up-and-go test was independently associated with lower values of ViPCr (p = 0.008). For all subjects and in the 8 patients unable to perform this test, ViPCr was negatively correlated with the ADL score (respectively p < 0.001 and p = 0.025). CONCLUSION: particularly in hospitalized and frail older subjects, muscle mitochondrial function was related to the global physical functional assessment.

Gadolinium-enhanced small-animal TOF magnetic resonance angiography.

So far, magnetic resonance angiography (MRA) of rodents has only been performed by using time-of-flight (TOF) MRI techniques. This is because applications of first-passage contrast agents as in humans are hampered by pronounced physiologic differences (blood volume and heart beat rate). Here we describe the use of low-dose Gd-DOTA to enhance the performance of TOF MRA in rat brain. While no improvement in contrast was achieved, the measuring time could be reduced by almost a factor of three. This decrease in total acquisition time has been used to study the impact of a model of ligatured common carotid on the upper part of the blood system of the rat.

Effect of creatine supplementation on phosphocreatine resynthesis, inorganic phosphate accumulation and pH during intermittent maximal exercise.

In this study, we examined the effect of creatine ingestion on muscle power output, muscle phosphocreatine resynthesis, inorganic phosphate and pH during repeated brief bouts of maximal exercise. Nine healthy males performed maximal plantar flexion before and after creatine ingestion (20 g x day(-1) for 6 days). The experimental protocol consisted of five 8 s bouts (bouts 1-5) interspersed with 30 s recovery, followed by bouts 6 (8 s) and 7 (16 s) separated by 1 and 2 min, respectively. Muscle phosphocreatine, inorganic phosphate and pH were estimated every 16 s by 31P magnetic resonance spectroscopy. After creatine ingestion, muscle power output increased by approximately 5% (P< 0.05) from bouts 3 to 7 and muscle phosphocreatine resynthesis increased (P< 0.05) during 10 min recovery. The higher phosphocreatine concentration observed after only 30 s of recovery was accompanied by lower inorganic phosphate accumulation and higher pH. Strong correlations were found between exercise power restoration and the corresponding pre-exercise phosphocreatine and inorganic phosphate concentrations and muscle pH after creatine ingestion. The better maintenance of muscle power output observed after creatine ingestion was attributed to a higher rate of phosphocreatine resynthesis, lower accumulation of inorganic phosphate and higher pH.

Dynamics and orientation of amphipathic peptides in solution and bound to membranes: a steady-state and time-resolved fluorescence study of staphylococcal delta-toxin and its synthetic analogues.

The environment of both the hydrophilic and hydrophobic sides of alpha-helical delta-toxin are probed by tryptophanyl (Trp) fluorescence, when self-association occurs in solution and on binding to membranes. The fluorescence parameters of staphylococcal delta-toxin (Trp15 on the polar side of the amphipathic helix) and synthetic analogues with single Trp at position 5 or 16 (on the apolar side) were studied. The time-resolved fluorescence decays of the peptides in solution show that the local environment of their single Trp is always heterogeneous. Although the self-association degree increases with concentration, as shown by fluorescence anisotropy decays, the lifetimes (and their statistical weight) of Trp16 do not change, contrary to what is observed for Trp15. The first step of self-association is then driven by hydrophobic interactions between apolar sides of alpha-helices, whilst further oligomerization involves their polar side (Trp15) via electrostatic interactions. This is supported by dissociation induced by salt. For all self-associated peptides, the polarity of the Trp microenvironment was not significantly modified upon binding to phospholipid vesicles, as indicated by the small shifts of the fluorescence emission spectra and lifetime values. However, the relative populations of the lifetime classes vary with bound-peptide density similar to the rates of their global motions in bilayers or smaller particles. Quenching experiments by water or lipid-soluble compounds show changes of the orientation of membrane-inserted peptides, from probably dimers lying flat at the interface at low peptide density, to oligomers spanning the membrane and inducing membrane fragmentation at high peptide density.

Functional and metabolic early changes in calf muscle occurring during nutritional repletion in malnourished elderly patients.

BACKGROUND: Metabolic alterations in skeletal muscle associated with malnutrition and the potential reversibility of such alterations during refeeding are not fully understood. OBJECTIVE: We characterized early changes in muscle during refeeding in malnourished, hospitalized elderly subjects. DESIGN: Muscle function, metabolism, and mass were evaluated in 24 clinically stable patients (11 were malnourished) by using isokinetic plantar flexor torque measurements and nuclear magnetic resonance (NMR) imaging for medial gastrocnemius mass assessment and 31P and 13C NMR spectroscopy for inorganic phosphate (Pi), phosphocreatine, and glycogen quantitation. RESULTS: Malnourished subjects had lower muscle mass (P < 0.02) and tended to have lower strength than did control subjects. In malnourished subjects, muscle strength increased after refeeding (P < 0.01) whereas muscle mass was unchanged. The ratio of Pi to ATP was lower in malnourished than in control subjects (P < 0.001) and increased during refeeding (P < 0.01). The mean ratio of phosphocreatine to ATP was lower in malnourished than in control subjects (P < 0.01) and increased to control values after refeeding. Muscle glycogen showed a scattered distribution for malnourished subjects; the mean value did not differ significantly from that of control subjects, either at baseline or after refeeding. CONCLUSIONS: The lower ratio of phosphocreatine to ATP in malnourished subjects could have resulted from either lower total muscle creatine or reduced oxidative capacities. High or normal glycogen associated with a low Pi-to-ATP ratio in malnourished subjects suggested preferential use of lipid over carbohydrate for energy supply, which is known to reduce muscle performance. The data suggest that normalization of muscle metabolite content after refeeding improves muscle strength in malnourished subjects.

Metabolism of [1-(13)C)glucose and [2-(13)C]acetate in the hypoxic rat brain..

The effects of hypoxia on the metabolism of the central nervous system were investigated in rats submitted to a low oxygen atmosphere (8% O(2); 92% N(2)). [1-(13)C]glucose and [2-(13)C]acetate were used as substrates, this latter being preferentially metabolized by glial cells. After 1-h substrate infusion, the incorporation of 13C in brain metabolites was determined by NMR spectroscopy. Under hypoxia, an important hyperglycemia was noted. As a consequence, when using labeled glucose, the specific enrichment of brain glucose C1 was lower (48.2+/-5.1%) than under normoxia (66.9+/-2.5%). However, relative to this specific enrichment, the (13)C incorporation in amino acids was increased under hypoxia. This suggested primarily a decreased exchange between blood and brain lactate. The glutamate C2/C4 enrichment ratio was higher under hypoxia (0.62+/-0.01) than normoxia (0.51+/-0.06), indicating a lower glutamate turnover relative to the neuronal TCA cycle activity. The glutamine C2/C4 enrichment ratio was also higher under hypoxia (0.87+/-0.07 instead of 0.65+/-0.11), indicating a new balance in the contributions of different carbon sources at the acetyl-CoA level. When using [2-(13)C]acetate as substrate, no difference in glutamine enrichment appeared under hypoxia, whereas a significant decrease in glutamate, aspartate, alanine and lactate enrichments was noted. This indicated a lower trafficking between astrocytes and neurons and a reduced tricarboxylic acid cycle intermediate recycling of pyruvate.

Single-coil surface imaging using a radiofrequency field gradient

A method for in-plane imaging of large objects as compared to the RF coil is proposed based on the use of a single specially designed surface coil, without using B(0) gradients. A constant B(1) gradient was generated along the main axis of a ladder-shaped coil, and RF-encoding along the direction of the gradient made it possible to obtain spin-density profiles. Successive acquisitions of profiles obtained by translation of the NMR coil resulted in distorted images-due to the presence of non-zero gradients perpendicular to the constant gradient-that were successfully processed using a mathematical treatment based on linear combinations of calculated altered images from single-pixel objects. Copyright 2000 Academic Press.

Aminomethylphosphonate and 2-aminoethylphosphonate as (31)P-NMR pH markers for extracellular and cytosolic spaces in the isolated perfused rat liver.

Aminomethylphosphonate (NMePo) and 2-aminoethylphosphonate (NEthPo) were evaluated as alternative pH indicators in the isolated perfused rat liver using (31)P-nuclear magnetic resonance (NMR). NMePo did not distribute within cells and remained in the extracellular space. It exhibited pH titration with a low pK(a) value (5.35). This behaviour makes NMePo useful for extracellular volume or acidic pH determination. In contrast, NEthPo accumulated within cells without altering liver energetic steady state, evaluated from nucleosides triphosphates resonances, even for prolonged (100 min) experiments. Withdrawal of NEthPo from perfusate revealed a residual resonance corresponding to the internalized amount of this phosphonate. This fraction was almost stable vs time and allowed determination of spin-lattice relaxation time constant T(1) within the liver (2.2 +/- 0.3 s; n = 6). Comparison of the titration curves for NEthPo and inorganic phosphate revealed that the accuracy of pH determination within physiologic or acidic range in both cases was comparable. Finally, when extracellular pH was decreased, the NEthPo resonance frequency was found to undergo the same chemical shift variations as observed for cytosolic P(i) signal, which was in good agreement with a cytosolic accumulation of this phosphonate. Therefore, NEthPo could be considered as an interesting cytosolic pH probe suitable for (31)P-NMR measurements, especially when experimental conditions prevent reliable observation of cytosolic Pi resonance.

The metabolism of [3-(13)C]lactate in the rat brain is specific of a pyruvate carboxylase-deprived compartment.

Lactate metabolism in the adult rat brain was investigated in relation with the concept of lactate trafficking between astrocytes and neurons. Wistar rats were infused intravenously with a solution containing either [3-(13)C]lactate (534 mM) or both glucose (750 mM) and [3-(13)C]lactate (534 mM). The time courses of both the concentration and (13)C enrichment of blood glucose and lactate were determined. The data indicated the occurrence of [3-(13)C]lactate recycling through liver gluconeogenesis. The yield of glucose labeling was, however, reduced when using the glucose-containing infusate. After a 20-min or 1-h infusion, perchloric acid extracts of the brain tissue were prepared and subsequently analyzed by (13)C- and (1)H-observed/(13)C-edited NMR spectroscopy. The (13)C labeling of amino acids indicated that [3-(13)C]lactate was metabolized in the brain. Based on the alanine C3 enrichment, lactate contribution to brain metabolism amounted to 35% under the most favorable conditions used. By contrast with what happens with [1-(13)C]glucose metabolism, no difference in glutamine C2 and C3 labeling was evidenced, indicating that lactate was metabolized in a compartment deprived of pyruvate carboxylase activity. This result confirms, for the first time from an in vivo study, that lactate is more specifically a neuronal substrate.

Liposomes as fatty acids carriers in isolated rat liver: effect on energy metabolism and on isolated mitochondria activity.

The effects of fatty acids (FA)-carrier, egg-lecithin liposomes (LIPO) as alternative to BSA, on ATP, glycogen and glucose contents in isolated perfused liver of fed rats were non-invasively studied using 31P/13C nuclear magnetic resonance (NMR). Oxidative phosphorylation was studied in isolated mitochondria from the same liver consecutively to the NMR experiments. ATP content decreased slowly and ATP turnover was similar during the perfusion with saline solution (KHB) or LIPO. However, LIPO induced an enhancement of respiratory control ratio in isolated mitochondria. Tissue glycogen and glucose content decreased when FA (linoleate or linolenate) were perfused with defatted BSA (3%) or LIPO (600 mg/l) whereas glucose excretion level was unchanged and lactate excretion tended to increase, reflecting changes in the cytosolic redox state and/or an enhancement of glycolysis. Addition of FA (0.5 or 1.5 mM) to LIPO caused a dramatic fall in liver ATP, a mitochondrial uncoupling and an impairment of the phosphorylation activity. Perfusion with FA (1.5 mM) carried by BSA significantly increased the ATP degradation without change of mitochondrial function. Owing to the higher affinity of BSA than LIPO for FA, these latter could be more easily released from complex LIPO-FA, increasing their uncoupling effect. Hence, the FA concentrations have to be largely decreased from the above currently used concentrations to avoid this effect. It will then be possible to minimize the effector action of FA and to study their more specific metabolic function as fuel. It was concluded that LIPO were appropriate carriers to study the different metabolic effects of FA.

13C nuclear magnetic resonance study of glycogen resynthesis in muscle after glycogen-depleting exercise in healthy men receiving an infusion of lipid emulsion.

In healthy individuals, glycogen recovery after a strong depletion is known to be rapid and insulin independent during the initial phase, and subsequently, slow and insulin dependent. Free fatty acids (FFAs) as a putative source of insulin resistance (IR) could thus impair glycogen recovery during the second period. Using in vivo 13C nuclear magnetic resonance (NMR), we studied the effect of long-chain triglyceride emulsion on gastrocnemius glycogen resynthesis during a 3-h recovery period after 90 min of moderate exercise consisting of plantar flexion on overnight-fasted healthy men (n = 8). In separate experiments, each subject was infused with 10% Ivelip (0.015 ml x kg(-1) x min(-1)) or 10% glycerol (0.13 mg x kg(-1) x min(-1)). NMR spectra were acquired before and at the end of the exercise and during the recovery period. Whole-body glucose and lipid oxidation rates (indirect calorimetry), plasma insulin, C-peptide, glucose, lactate, beta-hydroxybutyrate, triglycerides, and FFAs were determined. Glycogen consumption was 47.6 +/- 4.5% (glycerol) and 49.7 +/- 4.8% (Ivelip) of the initial glycogen. An acquired IR in the Ivelip group was significant at the onset of the recovery period by homeostasis model assessment (P = 0.002). Glycogen resynthesis in the glycerol group appeared faster during the 1st h than during the subsequent 2nd h of the postexercise period. The glycogen resynthesis level was significantly lower in the Ivelip group than in the glycerol group during the recovery period (P = 0.04 during the 1st h and P = 0.001 during the next 2 h). During the recovery, plasma lactate and whole-body oxidation rates were similar in the two groups, whereas glycemia was significantly higher in the Ivelip group. A decreased cellular uptake of glucose as a substrate for glycogenosynthesis, rather than a competition between oxidation of carbohydrate and FFA, is discussed.

Localised proton magnetic resonance spectroscopy of the brain after perinatal hypoxia: a preliminary report.

OBJECTIVES: Perinatal hypoxic ischaemic injury is a significant cause of neurodevelopmental impairment. The aim of this study was to evaluate localised proton magnetic resonance spectroscopy (1H-MRS) after birth asphyxia. MATERIALS AND METHODS: Thirty newborn infants suspected of having perinatal asphyxia (Apgar score < 3) were studied. The mean gestational age was 37 weeks, mean age at the MR examination was 18 days and mean weight was 2.9 kg. A 1.5-T unit was used for imaging and spectroscopy. None of the babies had mechanically assisted ventilation. No sedation was used. Axial T1-weighted and T2-weighted images were obtained. 1H-MRS was recorded in a single voxel, localised in white matter, using a STEAM sequence. RESULTS: Image quality was good in 25 of 30 babies. 1H-MRS was performed in 19 of 30 subjects, with adequate quality in 16. Choline, creatine/phosphocreatine and N-acetylaspartate peaks and peak-area ratios were analysed. Lactate was detected in four infants. The N-acetylaspartate/choline ratio was lower in infants with an impaired neurological outcome, but the difference was not statistically significant. CONCLUSIONS: This study suggests that 1H-MRS may be useful for assessing cerebral metabolism in the neonate. A raised lactate level and decreased N-acetylaspartate/choline ratio may be predictive of a poor outcome. However, in our experience this method is limited by the difficulty in performing the examination during the first hours after birth in critically ill babies, the problems related to use of a monovoxel sequence, the dispersion of the ratios and the lack of determination of the absolute concentration of the metabolites.

In vivo macrophage activity imaging in the central nervous system detected by magnetic resonance.

Cell-specific imaging has been proposed to increase the potential of magnetic resonance imaging (MRI) for tissue analysis. The hypothezis of the present work was that following intravenous injection of ultra-small particle iron oxide, a contrast agent that accumulates in mononuclear phagocyte cells, macrophages with iron burden would be detectable by MRI within the central nervous system at sites of inflammatory cellular activity. In experimental autoimmune encephalomyelitis in Lewis rats (in which intense macrophage activity results from both hematogenous macrophages and activated microglia), lesions have been seen by MRI as low signal intensities related to magnetic susceptibility effects induced by iron particles. Electron microscopy has revealed the presence of such particles within the cytoplasm of cells that had the morphological aspect of macrophages. Macrophage activity imaging might increase MRI capability with regard to the in vivo pathophysiological aspects of central nervous system (CNS) diseases and might help in therapeutic trials in the numerous CNS diseases in which macrophages are involved.

Magnetization transfer imaging of rat brain under non-steady-state conditions. Contrast prediction using a binary spin-bath model and a super-lorentzian lineshape.

Magnetization transfer contrast imaging using turbo spin echo and continuous wave off-resonance irradiation was carried out on rat brain in vivo at 4.7 T. By systematically varying the off-resonance irradiation power and the offset-frequency, the signal intensities obtained under steady-state for both transverse and longitudinal magnetization were successfully analyzed with a simple binary spin-bath model taking into account a free water compartment and a pool of protons with restricted motions bearing a super-Lorentzian lineshape. Due to important RF power deposition, such experimental conditions are not practical for routine imaging on humans. An extension of the model was derived to describe the system for shorter off-resonance pulse duration, i.e., when the longitudinal magnetization of the free protons has not reached a steady-state. Data sets obtained for three regions of interest, namely the corpus callosum, the basal ganglia, and the temporal lobe, were correctly interpreted for off-resonance pulse durations varying from 0.3 to 3 s. The parameter sets obtained from the calculations made it possible to predict the contrast between the different regions as a function of the pulse power, the offset frequency, and pulse duration. Such an approach could be extended to contrast prediction for human brain at 1.5 T.

Magnetization transfer fast imaging of implanted glioma in the rat brain at 4.7 T: interpretation using a binary spin-bath model.

C6 glioma cells were implanted in the left caudate nucleus of the rat brain. Histologic studies confirmed the presence of neoplastic tissue surrounded by a thin edematous region. Proton magnetization transfer contrast (MTC) fast imaging, using continuous wave off-resonance irradiation, was performed in vivo at 4.7 T with the rapid acquisition with relaxation enhancement (RARE) sequence. The observed MTC allowed very clear distinction of the tumoral region, in which magnetization transfer (MT) ratios were lower than in healthy tissues. Contrasts were analyzed as a function of the offset frequency and the amplitude of the radiofrequency (RF) irradiation. The contrast was higher between the contralateral basal ganglia and the tumor and lower between the tumor and the temporal lobe. Modeling of MT in the three brain regions was performed using a system including free water and a pool of protons with restricted motions. The rate of exchange between the two pools exhibited a decreasing hierarchy from the basal ganglia to the tumor. T2B values for the immobile protons ranged from 9.3 microsec in the basal ganglia to 7.5 microsec for the glioma. The acquisition conditions leading to the highest contrasts between the tumor and the healthy tissues correspond to 3,000 Hz offset frequency and 300 to 700 Hz RF irradiation amplitude.

Magnetization transfer imaging in vivo of the rat brain at 4.7 T: interpretation using a binary spin-bath model with a superLorentzian lineshape.

Proton magnetization transfer contrast (MTC) imaging, using continuous wave off-resonance irradiation, was performed on the rat brain in vivo at 4.7 Tesla. The observed MTC was studied in three different brain regions: the corpus callosum, the basal ganglia, and the temporal lobe. By systematically varying the offset frequency and the amplitude of the RF irradiation, the observed signal intensities for each region of interest were modeled using a system including free water and a pool of protons with restricted motions (R. M. Henkelman, X. Huang, Q. Xiang, G. J. Stanisz, SD Swanson, M. J. Bronskill, Magn. Res. Med. 29, 759 (1993)). Most of the relaxation parameters of both proton pools remained fairly constant for the three regions of interest, with a T2 value of about 9 micros for the immobilized protons, whereas the rate of exchange increased significantly from the temporal lobe to the corpus callosum. The optimal acquisition parameters for the improved MTC under steady-state saturation were found to be 2-10 kHz offset frequency and 500-800 Hz RF irradiation amplitude. Conversely, an irradiation amplitude of 3 kHz at an offset frequency of 12 kHz is required to minimize the direct effect of off-resonance irradiation. Such an approach could be extended to human brain imaging with the aim of characterizing tissue-specific disease.

Temperature dependence of NMR relaxation times of nucleoside triphosphates and inorganic phosphate in the isolated perfused rat liver. Effect on Pi compartmentation.

The effect of temperature on 31P NMR spectra from isolated perfused rat livers was studied at 9.4 T. Relaxation times (T1 and T2) of uncleoside triphosphates (NTP) and inorganic phosphate (Pi) were determined at 37, 25, 15, and 4 degrees C. Under hypothermic conditions, an unexpected apparent line sharpening in the Pi spectral region and a clear emergence of an additional Pi resonance were observed. This additional signal was assigned to mitochondrial Pi. T1 values obtained for cytosolic and mitochondrial Pi at 4 degrees C were 1.14 +/- 0.24 s (n = 5) and 0.71 +/- 0.18.s (n = 5), respectively. No significant mitochondrial contribution to the Pi resonance was observed at 37 degrees C. Quantification of Pi and NTP liver contents at 37 and 4 degrees C was performed by comparing the perfused liver spectrum and the corresponding perchloric acid extract spectrum. Under experimental conditions of low external Pi (0.12 mM), it was concluded that intracellular Pi was completely NMR-visible at 4 and 37 degrees C. The observation of the mitochondrial Pi signal at 4 degrees C was well explained by an increase in the Pi level within the matrix, in response to the mitochondrial swelling induced by hypothermia, as observed by electron microscopy. T2 values for the cytosolic Pi at 37 and 4 degrees C were 17 +/- 4 ms (n = 8) and 22 +/- 4 ms (n = 10), respectively. Comparison with measured linewidths indicated that line broadening for the main phosphorylated metabolites--including matrix Pi--was the result of B0 field inhomogeneity. The additional broadening of the cytosolic Pi resonance at 4 and 37 degrees C was attributed to pH heterogeneity within the liver.

31P magnetic resonance spectroscopy of human liver in elderly patients: changes according to nutritional status and inflammatory state.

Magnetic resonance spectroscopy (MRS) was used to determine the phosphorylated metabolite content in the liver of elderly patients in various nutritional states: normal, with protein deprivation, and with acute inflammatory syndrome. 31P-MRS investigations were performed at 1.5 T, and localized liver spectra were recorded using a two-dimensional chemical shift imaging sequence. Comparison to control spectra recorded on 10 healthy volunteers (age, 30.5 +/- 2.1 years) showed that the aging process does not significantly modify 31P-MRS liver spectra. Patients with protein deprivation exhibited a higher value than controls for the phosphomonoesters/nucleoside triphosphates (PME/NTP) ratio (P < .05). This increase was not due to the decrease of NTP, since the ratio of inorganic phosphate to NTP (Pi/NTP) remained constant. A decrease in the phosphodiesters to NTP (PDE/NTP) ratio (P < .04) contributed to the observed increase in the PME/PDE ratio (P < .01). In contrast, no significant difference in 31P-MRS spectra was found between elderly patients with hypoalbuminemia associated with inflammatory syndrome and the control group. We conclude that elderly patients with protein deprivation displayed changes in the level of phosphorylated metabolites in the liver that were not observed in the case of inflammatory syndrome despite lower serum albumin (Alb) concentrations.

Structural characterization of membrane insertion of M13 procoat, M13 coat, and Pf3 coat proteins.

A new, simple, and efficient purification method has been developed for the extremely hydrophobic M13 procoat, M13 coat, and Pf3 coat proteins. Homogeneous preparations were obtained in 2-propanol/0.1% TFA, where M13 coat protein is found to be dissolved in a monomeric form, and the two other proteins as dimers or trimers. The conformations of these particular proteins in different environments have been determined by circular dichroism and infrared spectroscopy. In organic solvents, the proteins adopt a conformation with an average helix content of 90%. In lipid bilayers composed of phosphatidylcholine and phosphatidylglycerol lipids, the average helix content is 50% for M13 procoat protein, 60% for M13 coat protein, and 75% for Pf3 coat protein. The orientational order parameter S alpha of the protein helices in planar lipid bilayers have been determined by polarized infrared measurements in the amide I spectral range. The helices of the three proteins are oriented preferentially parallel to the membrane normal, with S alpha = 0.63 for M13 procoat protein, S alpha = 0.58 for Pf3 coat protein, and a distinctly higher value of S alpha = 0.81 for M13 coat protein.