Muscle-associated triglyceride measured by computed tomography and magnetic resonance spectroscopy

Objective: Muscle triglyceride can be assessed in vivo using computed tomography (CT) and ¹H magnetic resonance spectroscopy (MRS), two techniques that are based on entirely different biophysical principles. Little is known, however, about the cross‐correlation between these techniques and their test—retest reliability. Research Methods and Procedures: We compared mean muscle attenuation (MA) in soleus and tibialis anterior (TA) muscles measured by CT with intra‐ and extramyocellular lipids (IMCL and EMCL, respectively) measured by MRS in 51 volunteers (26 to 72 years of age, BMI = 25.5 to 39.3 kg/m²). MA of midthighs was also measured in a subset (n = 19). Test—retest measurements were performed by CT (n = 6) and MRS (n = 10) in separate sets of volunteers. Results: MA of soleus was significantly associated with IMCL (r = ?0.64) and EMCL, which by multiple regression analysis was explained mostly by IMCL (p < 0.001) rather than EMCL (β = ?0.010, p = 0.94). Muscle triglyc‐eride was lower in TA than in soleus, and MA of TA was significantly correlated with EMCL (r = ?0.40) but not IMCL (r = ?0.16). By CT, MA of midthighs was correlated with MA in soleus (r = 0.40, p = 0.07) and whole calf (r = 0.62, p < 0.05). Finally, both MA and IMCL were highly reliable in soleus (coefficient of variation = <2% and 6.7%, respectively) and less reliable in TA (4% and 10%, respectively). Discussion: These results support the use of both CT and MRS as reliable methods for assessing skeletal muscle lipid.     

Fractal dimension in human cerebellum measured by magnetic resonance imaging

Fractal dimension has been used to quantify the structures of a wide range of objects in biology and medicine. We measured fractal dimension of human cerebellum (CB) in magnetic resonance images of 24 healthy young subjects (12 men and 12 women). CB images were resampled to a series of image sets with different 3D resolutions. At each resolution, the skeleton of the CB white matter was obtained and the number of pixels belonging to the skeleton was determined. Fractal dimension of the CB skeleton was calculated using the box-counting method. The results indicated that the CB skeleton is a highly fractal structure, with a fractal dimension of 2.57 +/- 0.01. No significant difference in the CB fractal dimension was observed between men and women.     

Adipose tissue volume measured by magnetic resonance imaging and computerized tomography in rats

The primary purpose of this study was to investigate the viability of magnetic resonance imaging (MRI) as a means of measuring the body composition of rodents. To do so we compared adipose tissue (AT) volumes measured by MRI with those obtained by X-ray computerized tomography (CT) in a group of rats (n = 17) varying in weight (465-815 g) and percent body fat (5.4-31.1%), with the latter determined by chemical analysis. For both MRI and CT, AT volumes (cm3) per transverse slice (3-mm thickness, 21-mm centers) were determined using a computer-based image analysis system that permitted detailed comparisons of both visceral and subcutaneous AT depots. Total AT volumes were calculated using a linear interpolation of AT areas obtained on consecutive slices. Correlation coefficients between MRI and CT for visceral [r = 0.98, standard error of estimate (SEE) = 6.8 cm3], subcutaneous (r = 0.98, SEE = 6.5 cm3), and total AT volumes (r = 0.99, SEE = 9.0 cm3) were highly significant (P less than 0.001). Both MRI- and CT-predicted AT mass (assuming fat density = 0.90 g/ml) correlated strongly with chemically extracted lipid (grams) values (r = 0.98, SEE 9.6 g and r = 0.99, SEE = 6.9 g, respectively). Post hoc Scheffé contrasts demonstrated that the mean AT and lipid mass values derived by the three methods were not significantly different (P = 0.01). No systematic differences were observed because the regression lines derived for either MRI or CT vs. chemical analysis were not significantly different from the identity line.(ABSTRACT TRUNCATED AT 250 WORDS)     

Stability and transmetallation of the magnetic resonance contrast agent MnDPDP measured by EPR

MnDPDP [manganese(II) N, N'-dipyridoxylethylenediamine- N, N'-diacetate-5,5'-bis(phosphate)] is the active component of Teslascan, a contrast medium for magnetic resonance imaging of the liver. It has previously been shown that MnDPDP is rapidly dephosphorylated to the monophosphate MnDPMP and the non-phosphorylated MnPLED, and that all these substances are rapidly transmetallated to the corresponding Zn complexes. In the present study we used EPR at 9 and 230 GHz to show that no free Mn(2+) ions can be detected in the product or in a mixture of MnDPDP and human serum. Competition experiments between MnDPDP and Zn(2+), Ca(2+), and Mg(2+) ions revealed approximately 15% transmetallation with Zn(2+) in a buffer system containing metal ion concentrations similar to that in serum, whereas approximately 10% transmetallation was obtained with Ca(2+) and only negligible transmetallation was obtained with Mg(2+) under these conditions. Binding experiments with Mn(2+) added to human albumin and human serum indicate that albumin accounts for most of the protein-bound Mn(2+) in serum.     

Water permeability ofNecturus gallbladder epithelial cell membranes measured by nuclear magnetic resonance

Summary In order to assess the contribution of transcellular water flow to isosmotic fluid transport acrossNecturus gallbladder epithelium, we have measured the water permeability of the epithelial cell membranes using a nuclear magnetic resonance method. Spin-lattice (T ₁) relaxation of water protons in samples of gallbladder tissue where the extracellular fluid contained 10 to 20mm Mn²⁺ showed two exponential components. The fraction of the total water population responsible for the slower of the two was 24±2%. Both the size of the slow component, and the fact that it disappeared when the epithelial layer was removed from the tissue, suggest that it was due to water efflux from the epithelial cells. The rate constant of efflux was estimated to be 15.6±1.0 sec¹ which would be consistent with a diffusive membrane water permeabilityP _d of 1.6×10³ cm sec¹ and an osmotic permeabilityP _os of between 0.3×10⁴ and 1.4×10⁴ cm sec¹ osmolar¹. Using these data and a modified version of the standing-gradient model, we have reassessed the adequacy of a fluid transport theory based purely on transcellular osmotic water flow. We find that the model accounts satisfactorily for near-isosmotic fluid transport by the unilateral gallbladder preparation, but a substantial serosal diffusion barrier has to be included in order to account for the transport of fluid against opposing osmotic gradients.     

Pressure effects on dipalmitoylphosphatidylcholine bilayers measured by 2H nuclear magnetic resonance

The effects of pressure, up to 5 kbar, on multilamellar vesicles of 1,2-dipalmitoyl-sn-phosphatidylcholine perdeuterated in the acyl chains (DPPC-d62) were examined by using high-pressure NMR techniques. A deuterium probe was built, and the quadrupole splitting was measured against pressure at various temperatures. The experiments were performed on pure lipid bilayers in the liquid-crystalline state and on bilayers in the liquid-crystalline state containing the local anesthetic tetracaine. The results show that the order parameter of all segments of the acyl chains increases with pressure in the liquid-crystalline state. The more highly ordered regions of the chains are affected slightly more than the regions near the methyl ends. The addition of tetracaine increases the disorder of the chains, and pressure reverses the effect of anesthetic on the lipid as seen by the reversal of the changes in line shape and the measured order parameter.     

Paediatric brain tissue properties measured with magnetic resonance elastography

Biomechanics and Modeling in Mechanobiology - The aim of this study is to characterise the stiffness of white and grey matter in paediatric subjects using magnetic resonance elastography (MRE) and...     

Amide proton exchange in human metallothionein-2 measured by nuclear magnetic resonance spectroscopy

In human metallothionein-2, the exchange rate constants of ten amide protons were found to range from 1.7 x 10(-4) to 1 x 10(-1) min-1 at pH 6.3 and 8 degrees C. Most of these slowly exchanging protons could be associated with hydrogen bonds in secondary structure elements of the alpha-domain. Amide proton exchange rates thus present an additional criterion for the structural characterization of different metallothioneins, which could be particularly valuable for comparisons of different homologous protein preparations containing nuclear magnetic resonance-inactive metal ions, where the metal-polypeptide co-ordinative bonds cannot be identified directly.     

Effects of age on pulmonary perfusion heterogeneity measured by magnetic resonance imaging.

Normal aging is associated with a decline in pulmonary function and efficiency of gas exchange, although the effects on the spatial distribution of pulmonary perfusion are poorly understood. We hypothesized that spatial pulmonary perfusion heterogeneity would increase with increasing age. Fifty-six healthy, nonsmoking subjects (ages 21-76 yr) underwent magnetic resonance imaging with arterial spin labeling (ASL) using a Vision 1.5-T whole body scanner (Siemens Medical Systems, Erlangen, Germany). ASL uses a magnetically tagged bolus to generate perfusion maps where signal intensity is proportional to regional pulmonary perfusion. The spatial heterogeneity of pulmonary blood flow was quantified by the relative dispersion (RD = SD/mean, a global index of heterogeneity) of signal intensity for voxels within the right lung and by the fractal dimension (D(s)). There were no significant sex differences for RD (P = 0.81) or D(s) (P = 0.43) when age was considered as a covariate. RD increased significantly with increasing age by approximately 0.1/decade until age 50-59 yr, and there was a significant positive relationship between RD and age (R = 0.48, P < 0.0005) and height (R = 0.39, P < 0.01), but not body mass index (R = 0.07, P = 0.67). Age and height combined in a multiple regression were significantly related to RD (R = 0.66, P < 0.0001). There was no significant relationship between RD and spirometry or arterial oxygen saturation. D(s) was not related to age, height, spirometry, or arterial oxygen saturation. The lack of relationship between age and D(s) argues against an intrinsic alteration in the pulmonary vascular branching with age as being responsible for the observed increase in global spatial perfusion heterogeneity measured by the RD.     

Anisotropic mechanical properties of magnetically aligned fibrin gels measured by magnetic resonance elastography

The anisotropic mechanical properties of magnetically aligned fibrin gels were measured by magnetic resonance elastography (MRE) and by a standard mechanical test: unconfined compression. Soft anisotropic biomaterials are notoriously difficult to characterize, especially in vivo. MRE is well-suited for efficient, non-invasive, and non-destructive assessment of shear modulus. Direction-dependent differences in shear modulus were found to be statistically significant for gels polymerized at magnetic fields of 11.7 and 4.7 T compared to control gels. Mechanical anisotropy was greater in the gels polymerized at the higher magnetic field. These observations were consistent with results from unconfined compression tests. Analysis of confocal microscopy images of gels showed measurable alignment of fibrils in gels polymerized at 11.7 T. This study provides direct, quantitative measurements of the anisotropy in mechanical properties that accompanies fibril alignment in fibrin gels.     

Quantitative and noninvasive assessment of prenatal X-ray-induced CNS abnormalities using magnetic resonance imaging

Our purpose was to noninvasively assess formation of the microvasculature, blood-brain barrier (BBB) and blood-CSF barrier formation of prenatal X-ray-induced CNS abnormalities using quantitative MRI. Eight pregnant female Sprague-Dawley rats were divided into two groups consisting of control and X-irradiated animals. After birth, 20 neonatal male rats were divided into four groups of five rats. To evaluate the development of the BBB, changes in T(1) induced by Gd-DTPA were compared quantitatively in normal and prenatally irradiated animals in the formative period 1 to 2 weeks after birth. To assess the abnormalities of the microvasculature, quantitative perfusion MRI and MR angiography were also used. Histology was also performed to evaluate the BBB (albumin) and vascular endothelial cells (laminin). Decreased cerebral blood flow (CBF) and angioarchitectonic abnormalities were observed in the prenatally irradiated rats. However, abnormalities of the BBB and blood-CSF barrier were not observed using Gd-enhanced MRI and albumin staining. Quantitative perfusion MRI, MR angiography and Gd-enhanced T(1) mapping are useful for assessing CNS disturbance after prenatal exposure to radiation. These techniques provide important diagnostic information for assessing the condition of patients during the early stages of life after accidental or unavoidable prenatal exposure to radiation.     

Sodium binding in Halobacterium halobium measured by the nuclear magnetic resonance technique

Relaxation time measurements T₁ and T₂ of sodium in Halobacterium halobium pellets were carried out at two frequencies. From those measurements, combined with intensity measurements of the sodium in the system, estimation of the properties of the sodium ions in the system was carried out. It is suggested that three types of sodium ions are present in the bacterial pellet. (A) The extracellular sodium with properties of free solution and (B) sodium which is in the pericellular volume between the cell wall and the cell membrane. There is an exchange between type A and type B sodium. The type B sodium has (e²qQ)/h = 3.7 · 10⁷ rad/s, τ_cB = 5.2 · 10⁻⁶ s and τ_B = 1 · 10⁻³ s. The sodium of type C is bound inside the cell and undetected. It's concentration inside the cell is assumed to be 1.9 M.     

Monitoring histone deacetylase inhibition in vivo: noninvasive magnetic resonance spectroscopy method

Histone deacetylase inhibitors (HDACis) are emerging as promising and selective antitumor agents. However, HDACis can lead to tumor stasis rather than shrinkage, in which case, traditional imaging methods are not adequate to monitor response. Consequently, novel approaches are needed. We have shown in cells that (19)F magnetic resonance spectroscopy (MRS)-detectable levels of the HDAC substrate Boc-Lys-TFA-OH (BLT) are inversely correlated with HDAC activity. We extended our investigations to a tumor xenograft model. Following intraperitoneal injection of BLT, its accumulation within the tumor was monitored by in vivo (19)F MRS. In animals treated with the HDACi suberoylanilide hydroxamic acid (SAHA), tumoral BLT levels were higher by 77% and 132% on days 2 and 7 of treatment compared with pretreatment levels (n = 6; p < .05). In contrast, tumoral BLT levels remained unchanged in control animals and in normal tissue. Thus, (19)F MRS of BLT detected the effect of HDACi treatment as early as day 2 of treatment. Importantly, tumor size confirmed that SAHA treatment leads to inhibition of tumor growth. However, difference in tumor size reached significance only on day 6 of treatment. Thus, this work identifies BLT as a potential molecular imaging agent for the early noninvasive MRS detection of HDAC inhibition in vivo.     

Study of agreement between LDL size as measured by nuclear magnetic resonance and gradient gel electrophoresis

LDL particle size can be measured by gradient gel electrophoresis (GGE) and NMR. The agreement between the two methods has not been extensively evaluated. Therefore, we measured LDL size by NMR and GGE in 324 individuals (152 with type 1 diabetes and 172 controls). The Spearman correlation between both methods was 0.39 [95% confidence interval (CI) = 0.29, 0.48]. The average difference was 5.38 nm (NMR being smaller), but it increased with increasing LDL size. Less than 50% of people classified as pattern B on GGE were classified as pattern B on NMR (kappa = 0.31; 95% CI = 0.17, 0.45). Agreement was lower for diabetic subjects compared with controls, for women compared with men, and for subjects with triglycerides less than 1.30 mmol/l compared with subjects with triglycerides greater than 1.30 mmol/l. External validation showed that cholesteryl ester transfer rate was related to LDL size on GGE in all subgroups and to LDL size on NMR only in men and nondiabetic subjects. Our findings show that agreement between NMR- and GGE-based LDL size is far from perfect and is not consistent across subgroups of patients. In particular, the two methods should not be assumed to be interchangeable in women and diabetic subjects. Whether NMR or GGE predicts cardiovascular disease risk better has not yet been evaluated.     

Water permeability of chlorella cell membranes by nuclear magnetic resonance: measured diffusion coefficients and relaxation times

Measurement by two nuclear magnetic resonance (NMR) techniques of the mean residence time τ_a of water molecules inside Chlorella vulgaris (Beijerinck) var. “viridis” (Chodot) is reported. The first is the Conlon and Outhred (1972 Biochim Biophys Acta 288: 354-361) technique in which extracellular water is doped with paramagnetic Mn²⁺ ions. Some complications in application of this technique are identified as being caused by the affinity of Chlorella cell walls for Mn²⁺ ions which shortens the NMR relaxation times of intra- and extracellular water. The second is based upon observations of effects of diffusion on the spin echo of intra- and extracellular water. Echo attenuation of intracellular water is distinguished from that of extracellular water by the extent to which diffusive motion is restricted. Intracellular water, being restricted to the cell volume, suffers less echo attenuation. From the dependence of echo amplitude upon gradient strength at several values of echo time, the mean residence time of intracellular water can be determined. From the mean residence time of intracellular water, the diffusional water permeability coefficient of the Chlorella membrane is calculated to be 2.1 ± 0.4 × 10⁻³ cm sec⁻¹.     

Ethanol transport in Zymomonas mobilis measured by using in vivo nuclear magnetic resonance spin transfer.

For the first time, unidirectional rate constants of ethanol diffusion through the lipid membrane of a microorganism, the bacterium Zymomonas mobilis, were determined, thus replacing indirect inferences with direct kinetic data. The rate constants k1 (in to out) were 6.8 +/- 0.4s(-1) at 29 degrees C and 2.7 +/- 0.3s(-1) at 20 degrees C. They were determined by using 1H selective nuclear magnetic resonance spin magnetization transfer. The measurements were done on l-ml cell suspensions. No addition of radiotracers, withdrawing of aliquots, physical separation methods, or chemical manipulations were required. Until now, the rate constants of ethanol transport in microorganisms have been unknown because ethanol diffuses through the cytoplasmic membrane too quickly for radiolabel approaches. Net velocities of ethanol exchange were calculated from unidirectional rate constants and cytoplasmic volume, which was also determined with the same nuclear magnetic resonance experiments. The results (i) confirmed that ethanol would not be rate limiting during the conversion of glucose by Z. mobilis and (ii) indicated that ethanol can serve as an in vivo marker of cytoplasmic volume changes. This was verified by monitoring for the first time the changes of both cytoplasmic volume and extracytoplasmic and cytoplasmic concentrations of alpha and beta anomers of D-glucose in cell suspensions of a microorganism. These findings may open up new possibilities for kinetic studies of ethanol and sugar transport in Z. mobilis and other organisms.