1H NMR metabolic profiling of synovial fluid from patients with anterior cruciate ligament tears and hemarthrosis.

OBJECTIVE: To compare the metabolic profiles of synovial fluid (SF) from patients with anterior cruciate ligament tears and hemarthrosis (HA) with that of normal controls, using 1H NMR spectroscopy (NMRS). METHODS: Synovial fluid was collected from eleven patients undergoing arthroscopic debridement within 14 days following an anterior cruciate ligament (ACL) tear and hemarthrosis. Ten additional SF samples were obtained from the knees of osteoarthritis-free volunteers to serve as normal controls. The relative concentrations of twenty-eight endogenous SF metabolites (hydroxybutyrate, acetate, acetoacetate, acetone, alanine, arginine, choline, citrate, creatine, creatinine, formate, glucose, glutamate, glutamine, glycerol, glycine, histidine, isoleucine, lactate, leucine, lysine, phenylalanine, proline, pyruvate, threonine, tyrosine, valine, and the mobile components of glycoproteins and lipids) were evaluated using NMRS and quantified using CHENOMX metabolomics analysis software. Mean differences between groups were evaluated with t-tests controlling for multiple comparisons at an overall error rate of 0.10. RESULTS: Statistically significant increases in the levels of glucose, choline, the branched-chain amino acids leucine, isoleucine, and valine, and the mobile components of N-acetyl glycoproteins and lipids were observed in ACL/HA SF as compared with normal controls; lactate levels were reduced. CONCLUSIONS: Marked changes occur in the metabolic profiles of human knee fluid following ACL injury and hemarthrosis, suggestive of increased demand and accompanying inflammatory response; potentially increased lipid and glucose metabolism; and possible hyaluronan degradation within the joint following trauma.

Design and implementation of a 3D-MR/CT geometric image distortion phantom/analysis system for stereotactic radiosurgery.

The design, construction and application of a multimodality, 3D magnetic resonance/computed tomography (MR/CT) image distortion phantom and analysis system for stereotactic radiosurgery (SRS) is presented. The phantom is characterized by (1) a 1 × 1 × 1 (cm)3 MRI/CT-visible 3D-Cartesian grid; (2) 2002 grid vertices that are 3D-intersections of MR-/CT-visible 'lines' in all three orthogonal planes; (3) a 3D-grid that is MR-signal positive/CT-signal negative; (4) a vertex distribution sufficiently 'dense' to characterize geometrical parameters properly, and (5) a grid/vertex resolution consistent with SRS localization accuracy. When positioned correctly, successive 3D-vertex planes along any orthogonal axis of the phantom appear as 1 × 1 (cm)2-2D grids, whereas between vertex planes, images are defined by 1 × 1 (cm)2-2D arrays of signal points. Image distortion is evaluated using a centroid algorithm that automatically identifies the center of each 3D-intersection and then calculates the deviations dx, dy, dz and dr for each vertex point; the results are presented as a color-coded 2D or 3D distribution of deviations. The phantom components and 3D-grid are machined to sub-millimeter accuracy, making the device uniquely suited to SRS applications; as such, we present it here in a form adapted for use with a Leksell stereotactic frame. Imaging reproducibility was assessed via repeated phantom imaging across ten back-to-back scans; 80%-90% of the differences in vertex deviations dx, dy, dz and dr between successive 3 T MRI scans were found to be ⩽0.05 mm for both axial and coronal acquisitions, and over >95% of the differences were observed to be ⩽0.05 mm for repeated CT scans, clearly demonstrating excellent reproducibility. Applications of the 3D-phantom/analysis system are presented, using a 32-month time-course assessment of image distortion/gradient stability and statistical control chart for 1.5 T and 3 T GE TwinSpeed MRI systems.

Development of a geometrically accurate imaging protocol at 3 Tesla MRI for stereotactic radiosurgery treatment planning.

The purpose of this study is to develop a geometrically accurate imaging protocol at 3 T magnetic resonance imaging (MRI) for stereotactic radiosurgery (SRS) treatment planning. In order to achieve this purpose, a methodology is developed to investigate the geometric accuracy and stability of 3 T MRI for SRS in phantom and patient evaluations. Forty patients were enrolled on a prospective clinical trial. After frame placement prior to SRS, each patient underwent 3 T MRI after 1.5 T MRI and CT. MR imaging protocols included a T1-weighted gradient echo sequence and a T2-weighted spin echo sequence. Phantom imaging was performed on 3 T prior to patient imaging using the same set-up and imaging protocols. Geometric accuracy in patients and phantoms yielded comparable results for external fiducial reference deviations and internal landmarks between 3 T and 1.5 T MRI (mean ≤ 0.6 mm; standard deviation ≤ 0.3 mm). Mean stereotactic reference deviations between phantoms and patients correlated well (T1: R = 0.79; T2: R = 0.84). Statistical process control analysis on phantom QA data demonstrated the stability of our SRS imaging protocols, where the geometric accuracy of the 3 T SRS imaging protocol is operating within the appropriate tolerance. Our data provide evidence supporting the spatial validity of 3 T MRI for targeting SRS under imaging conditions investigated. We have developed a systematic approach to achieve confidence on the geometric integrity of a given imaging system/technique for clinical integration in SRS application.

Magnetic resonance imaging in the radiation treatment planning of localized prostate cancer using intra-prostatic fiducial markers for computed tomography co-registration.

PURPOSE: To assess the feasibility, and potential implications, of using intra-prostatic fiducial markers, rather than bony landmarks, for the co-registration of computed tomography (CT) and magnetic resonance (MR) images in the radiation treatment planning of localized prostate cancer. METHODS: All men treated with conformal therapy for localized prostate cancer underwent routine pre-treatment insertion of prostatic fiducial markers to assist with gross target volume (GTV) delineation and to identify prostate positioning during therapy. Six of these men were selected for investigation. Phantom MRI measurements were obtained to quantify image distortion, to determine the most suitable gold alloy marker composition, and to identify the spin-echo sequences that optimized both marker identification and the contrast between the prostate and the surrounding tissues. The GTV for each patient was contoured independently by three radiation oncologists on axial planning CT slices, and on axial MRI slices fused to the CT slices by matching the implanted fiducial markers. From each set of contours the scan common volume (SCV), and the scan encompassing volume (SEV), were obtained. The ratio SEV/SCV for a given scan is a measure of inter-observer variation in contouring. For each of the 18 patient-observer combinations the observer common volume (OCV) and the observer encompassing volume (OEV) was obtained. The ratio OEV/OCV for a given patient-observer combination is a measure of the inter-modality variation in contouring. The distance from the treatment planning isocenter to the prostate contours was measured and the discrepancy between the CT- and the MR-defined contour recorded. The discrepancies between the CT- and MR-defined contours of the posterior prostate were recorded in the sagittal plane at 1-cm intervals above and below the isocenter. RESULTS: Phantom measurements demonstrated trivial image distortion within the required field of view, and an 18K Au/Cu alloy to be the marker composition most suitable for CT-MRI image fusion purposes. Inter-observer variation in prostate contouring was significantly less for MR compared to CT. The mean SEV/SCV ratio was 1.58 (confidence interval (CI): 1.47-1.69) for CT scans and 1.37 (CI: 1.33-1.41) for MR scans (paired t-test; P=0.036). The overall magnitude of contoured GTV was similar for MR and CT; however, there were spatial discrepancies in contouring between the two modalities. The greatest systematic discrepancy was at the posterior apical prostate border, which was defined 3.6 mm (SD 3.5 mm) more posterior on MR- than CT-defined contouring. CONCLUSIONS: Prostate contouring on MR is associated with less inter-observer variation than on CT. In addition, we have demonstrated the feasibility of using intra-prostatic fiducial markers, rather than bony landmarks, for the co-registration of CT and MR images in the radiation treatment planning of localized prostate cancer. This technique, together with on-line correction of treatment set-up according to the fiducial marker position on electronic portal imaging, may enable a reduction in the planning target volume (PTV) margin needed to account for inter-observer error in target delineation, and for prostate motion.

Macromolecule and water magnetization exchange modeling in articular cartilage.

Magnetization exchange effects between the matrix macromolecules (e. g., collagen and proteoglycan) and water were examined in normal, deuterated, and proteoglycan-depleted articular cartilage. Relaxation results (T(2), T(1rho), and T(1)) suggested that a four-site exchange scheme provided an accurate model for articular cartilage relaxation and interspin group coupling details. Magnetization exchange within the collagen-bulk-water, proteoglycan-collagen, and collagen fibrillar water-collagen cartilage subsystems were quantified. Although collagen-bulk-water was the largest of the cartilage coupling subsystems ( approximately 90% signal) and is exploited in MRI, the rates of magnetization transfer (MT) within the latter subsystems were appreciably larger. Magnetization exchange rates for proteoglycan-collagen and collagen fibrillar water-collagen were 120 s(-1) and 4.4 s(-1), respectively. The observation of these latter two exchange subsystems suggested potential clinical MRI-MT applications in detecting molecular abnormalities associated with osteoarthritis.

Amelioration of disease severity by intraarticular hylan therapy in bilateral canine osteoarthritis.

Because of its high molecular weight, the glycosaminoglycan molecule hyaluronan is responsible for the viscoelastic properties of normal synovial fluid. In osteoarthritis, the concentration and molecular weight of hyaluronan in synovial fluid is diminished: this impairs the ability of synovial fluid to effectively lubricate joints, absorb loads, and exert anti-inflammatory effects. Using a bilateral anterior cruciate-ligament transection and partial neurectomy canine model of osteoarthritis, this study examined the effect of viscosupplementation with hylan G-F 20 as a treatment for osteoarthritis. Twelve dogs underwent bilateral arthroscopic anterior cruciate-ligament transections and partial neurectomy of the knee joints. Beginning 1 week after the operation, six dogs received three weekly 500-microl injections of hylan G-F 20 in one knee and a sham injection of saline solution in the contralateral knee (early-treatment group). The remaining six animals underwent the same treatment 2 months following the procedure (late-treatment group). All dogs were killed at 8 months, and both knees were evaluated for gross pathology, histology, and proteoglycan content. In addition, with use of 500-MHz [1H] magnetic resonance spectroscopy, the synovial fluid from both knees was assessed for changes in metabolic profile. Differences in outcome were analyzed with paired t tests. Gross pathological and histological examination revealed significantly less severe changes of osteoarthritis in knees treated with hylan G-F 20 2 months after surgery than in the contralateral untreated knees. Magnetic resonance spectroscopy of the specimens in this late-treatment group showed significantly decreased glucose concentrations and significantly elevated isoleucine levels in the synovial fluid from knees treated with hylan G-F 20 compared with the controls. Previous magnetic resonance spectroscopy had shown that glucose concentrations increase with the onset of osteoarthritis and eventually diminish in end-stage osteoarthritis. The three injections of hylan were given after osteoarthritis was established, and the severity of the disease was ameliorated in the treated knees 6 months after treatment. This occurred although hylan G-F 20 is almost certainly cleared from joints by lymphatics within 4 weeks of injection, suggesting that hylan therapy can retard the progression of osteoarthritis for periods of time extending beyond the intraarticular residence time of the injected molecules and that hylan injections given at relatively early stages of osteoarthritis may have a chondroprotective effect. No changes in outcome were noted in the animals that received hylan G-F 20 immediately following surgery.

Tunneling spectroscopy from magnetization evolution in a tilted rotating frame of nuclear spins.

The time evolution of the proton Zeeman magnetization in the rotating frame at the magic angle theta(M) = cos(-1)(1/3) is calculated for an isolated tunneling methyl group and its Fourier transform is given. The calculation compares well with the experimental spectra of CH(3)CD(2)I and methylmalonic acid. It is shown that Fourier transform spectroscopy of the magnetization evolution in a tilted RF frame represents an excellent alternative to the analogous experiment performed at exact resonance, resulting in improved resolution and a much better signal-to-noise ratio.

The effects of freeze/thawing on human synovial fluid observed by 500 MHz 1H magnetic resonance spectroscopy.

OBJECTIVE: To investigate the effect of freeze/thaw and low temperature storage on the biomolecular profile of human synovial fluid (SF) using high resolution (500 MHz 1H) magnetic resonance spectroscopy (MRS). METHODS: SF was collected from 12 patients undergoing arthroscopic debridement for treatment of moderate osteoarthritis (OA). Six of the larger samples were divided into 5 parts and treated as follows: the first was analyzed with spin-echo MRS soon after arthroscopy (< 24 h); the 2nd, 3rd, and 4th parts were frozen (-75 degrees C) and thawed for a total of one, 5, and 10 freeze/thaw cycles, respectively, followed by MRS analysis; the 5th part was kept in -75 degrees C storage for > or = 1 year before MRS processing. The 6 smaller samples were divided into 2 parts, the first analyzed shortly after extraction (< or = 24 h), while the 2nd was processed after storage at -75 degrees C for > or = 1 year. Changes in measured metabolite levels were tested for significance using paired t tests. RESULTS: Freeze-thaw cycling had no statistically significant effect on the relative concentrations of endogenous metabolites measured by MRS, though it did alter individual sample results. Prolonged low temperature storage resulted in a significant drop (p < 0.05) in the signal intensities of glucose (45%), N-acetyl glycoproteins (39%), CH2-chain and CH3-terminal and resonances of lipoproteins (46 and 37%, respectively), valine (43%), leucine (35%), and isoleucine (43%). CONCLUSION: This study raises questions about routine procedures that may inadvertently affect the outcomes of quantitative SF analyses. Extended low temperature storage should be avoided as it permanently alters the biochemical profile of SF, possibly leading to erroneous conclusions about the nature of OA related changes in metabolite levels with disease progression.

Design of a high-power NMR probe for low-temperature studies.

A low-temperature, high-power NMR probe head design is described which eliminates the problem of electric arc discharge commonly experienced during radiofrequency pulse cycling in a helium environment. A polychlorotrifluoroethylene (Kel-F) coil former, fitted with a solenoid coil, is heat-shrunk onto stainless-steel flanges and spot-welded inside a stainless-steel probe head assembly connected to a hollow coaxial transmission-line probe shaft. By this means, the sample coil and all high-voltage elements can effectively be isolated in a vacuum, while at the same time permitting good thermal contact between the sample and cryogenic gas. This design was used in NMR studies in the 4.6 K < or = T < or = 77 K temperature range for RF pulse durations < or = 50 ms (and longer for low RF amplitudes) and amplitudes up to approximately 60 G.

Spectroscopy from proton spin magnetization evolution in their rotating frame: A study of small tunneling splitting.

The time evolution of proton Zeeman magnetization in the rotating frame at exact resonance, omega = omega(0), is evaluated for an isolated tunneling methyl group CH(3). The Fourier transform of this evolution in time is calculated and both its real and imaginary components are presented. It is shown that the real component does not depend significantly on the strength of the preparation pulse when the tunneling splitting of the methyl rotator ground state is less than 100 kHz. It is also found that the imaginary component of the transform is inversely proportional to the strength of the preparation RF pulse. This is a consequence of the partial dephasing of proton spins during the preparation pulse. The results of the calculation compare well with the experimental spectra of CH(3)CD(2)I.

1H NMR investigation of changes in the metabolic profile of synovial fluid in bilateral canine osteoarthritis with unilateral joint denervation.

OBJECTIVE: High resolution 1H-nuclear magnetic resonance (NMR) techniques have been used to compare the effects of unilateral knee-joint denervation on the biochemical profiles of synovial fluid in a bilateral canine model of osteoarthritis. METHOD: Paired synovial fluid samples were obtained from seven dogs all of which had previously undergone bilateral anterior cruciate ligament transection, unilateral knee denervation and contralateral sham nerve exposure. All synovial fluid samples were then analyzed using 500 MHz 1H-CPMG spin-echo NMR Spectroscopy to assess differences in endogenous metabolite levels between the paired fluids. RESULTS: The results indicate statistically significant increases in glycerol, hydroxybutyrate, glutamine/glutamate, creatinine/creatine, acetate and N-acetyl-glycoprotein concentrations in synovial fluids from denervated with respect to control knees. Furthermore, significant trends towards elevated lactate, alanine and pyruvate levels in the denervated knee fluids are consistent with our previous findings comparing NMR spectroscopy metabolic profiles of normal and osteoarthritic canine synovial fluids. CONCLUSION: This study lends support to the principle of neurogenic acceleration of OA in that the observed differences in metabolite concentrations found in the denervated knee fluids seem to correlate with metabolic changes resulting from aggravation of the OA process caused by joint denervation.

Comparative study of normal and osteoarthritic canine synovial fluid using 500 MHz 1H magnetic resonance spectroscopy.

High resolution 1H nuclear magnetic resonance spectroscopy has been used to investigate and compare the metabolic profiles of normal and osteoarthritic synovial fluids in a canine model of osteoarthritis. The spectra of osteoarthritic synovial fluid showed (a) increased concentrations of lactate, pyruvate, lipoprotein-associated fatty acids, and glycerol as well as the ketones hydroxybutyrate and hydroxyisobutyrate, (b) reduced levels of glucose, and (c) elevated levels of N-acetylglycoproteins, acetate, and acetamide compared with healthy normal canine synovial fluid. An increase was also observed in the concentrations of the amino acids alanine and isoleucine. These results suggest that (a) the intraarticular environment in canine osteoarthritis is more hypoxic and acidotic than in a normal joint, (b) lipolysis may play an increasingly important role as a source of energy in osteoarthritis, and (c) the N-acetylglycoprotein polymer component of synovial fluid (mostly hyaluronan) seems to be increasingly fragmented and degraded into acetate by way of an acetamide intermediate with progressive osteoarthritis. The observed changes in the biochemical profile of canine osteoarthritic synovial fluid may be useful in understanding alterations in joint metabolism consequent to arthritic diseases and helpful in identifying potential markers of osteoarthritis.

31P-nuclear magnetic resonance studies of bioenergetic changes in skeletal muscle in malnourished human adults.

In previous studies, both animals and malnourished children receiving 25% of the protein-energy intake of a control group, resulting in a 25% weight loss, had lower ratios of phosphocreatine to beta-ATP and of phosphocreatine to inorganic phosphorus, higher free ADP concentrations, and lower free energy of ATP hydrolysis than the control group. Therefore, the effect of malnutrition on muscle energetics in adult humans was examined by using 31P-nuclear magnetic resonance spectroscopy in malnourished patients with a mean body mass index (BMI; in kg/m2) of 16.4 compared with healthy control subjects with a significantly higher body mass index of 24.5 (P < 0.005). The mean (+/- SEM) ratio of phosphocreatine (PCr) to ATP in the malnourished patients was 2.28 +/- 0.27, which was significantly lower than the ratio of 3.1 +/- 0.15 in control subjects (P < 0.02). The ratio of inorganic phosphorus (Pi) to ATP in malnourished patients was 0.33 +/- 0.04, which was significantly lower than the ratio of 0.48 +/- 0.03 in control subjects (P < 0.02), but the ratio of PCr to Pi was not significantly different from that in control subjects. There was a significant correlation between BMI and the ratio of PCr to ATP (P < 0.01) and of Pi to ATP (P < 0.01). These data suggest that progressive loss of BMI is associated with a relative loss of muscle creatine and phosphorus in relation to ATP. The findings were unlikely to have been due only to atrophy of fast-twitch fibers because such atrophy would have altered the ratio of PCr to Pi.