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F Eckstein

Publications and source records attributed to F Eckstein.

At least 73 records · Page 4Linked to original sources
PubMed2001

Optimization and validation of a rapid high-resolution T1-w 3D FLASH water excitation MRI sequence for the quantitative assessment of articular cartilage volume and thickness.

In view of follow up, survey and development of therapeutic strategies for osteoarthritis where cartilage deterioration plays an important role, a non invasive, reliable and quantitative assessment of the articular cartilage is desirable. The currently available high resolution T(1)-weighted (T1-w) 3D FLASH pulse sequences with frequency selective fat suppression are very time consuming. We have 1) optimized a high resolution T1-w 3D FLASH water excitation (WE) sequence for short acquisition time and cartilage visualization, and 2) validated this sequence for cartilage volume and thickness quantification. The spectral fat presaturation was replaced by selective water excitation. The flip angle of the WE sequence was optimized for the contrast to noise (C/N(cart)) ratio of cartilage. Sagittal datasets (voxel size: 0.31 x 0.31 x 2 mm(3)) of the knees of nine healthy volunteers were acquired both, with the 3D FLASH WE (17.2/6.6/30 degrees ) sequence (WE) and a previously validated 3D FLASH fat saturated (42/11/30 degrees ) sequence (FS). For validation of the WE sequence, cartilage volume, mean and maximal cartilage thickness of the two sequences were compared. Reproducibility was assessed by calculating the coefficient of variation (COV %) of 4 consecutive WE data sets in the volunteers. The acquisition time was reduced from 16'30" (FS) down to 7'14" for the WE sequence. Image contrast and visualization of the cartilage was very similar, but delineation of the basal layer of the cartilage was slightly improved with the WE sequence. A flip angle of 30 degrees provided the best C/N(cart) ratios (WE). Reproducibility (COV) was between 1.9 and 5.9%. Cartilage volume and thickness agreed within 4% between FS and WE sequence. The WE sequence allows for rapid, valid and reproducible quantification of articular cartilage volume and thickness, prerequisites for follow-up examinations. The reduced acquisition time (50% of FS) enables routine clinical application and thus may contribute to a broader assessment of osteoarthritis.

Adult↗
PubMed2001

Three-dimensional analysis of shoulder girdle and supraspinatus motion patterns in patients with impingement syndrome.

Alterations of the shoulder girdle motion have been suggested to be associated with shoulder disorders. The objective of this study was to perform a three-dimensional (3D) motion analysis of the supraspinatus muscle and shoulder girdle in patients with different stages of impingement syndrome. 20 patients with unilateral impingement and 14 normal controls were investigated at 30 degrees, 90 degrees, and 120 degrees of abduction with and without abducting muscle activity. The spatial relationship between the shoulder girdle elements and the supraspinatus was quantified from open MRI data. No significant alterations in glenoid rotation were observed between the patients and asymptomatic volunteers. However, while in the healthy volunteers the values showed a normal distribution (28.5+/-3.6 degrees at 90 degrees abduction with muscle activity), the patients (30.5 degrees+/-9.7 degrees) contained a subset of five individuals with an obvious increase in glenoid rotation angle (>40 degrees) compared with controls (>2.5 standard deviations higher than the mean) and with the healthy contralateral side. These five patients also displayed alterations in the scapulo-humeral rhythm and supraspinatus motion, but not in clavicular position. The study shows that only a specific subset of patients with impingement syndrome demonstrates complex changes in shoulder girdle and supraspinatus motion patterns, suggesting that this subset may benefit from an alternative type of treatment.

Adult↗
PubMed2001

Interindividual variability and correlation among morphological parameters of knee joint cartilage plates: analysis with three-dimensional MR imaging.

OBJECTIVE: To determine the range and variability of the cartilage volume, thickness, and articular surface areas in the knee joints of healthy male subjects, the association of these parameters within and between the knee joint cartilage plates, and their correlation with anthropometric variables. METHOD: The right knees of 27 individuals (age 23 to 64 years) without cartilage damage were examined. Sagittal magnetic resonance imaging was with a fat-suppressed gradient echo sequence (resolution 2 x 0.31 x 0.31 mm(3)), quantitative parameters being computed for all cartilage plates. RESULTS: The total knee joint cartilage volume ranged from 16.6 to 31.4 ml, the size of the articular surfaces from 102 to 163 cm(2), and the mean cartilage thickness from 1.57 to 2.43 mm. The mean and maximal cartilage thickness were highest in the patella (2.76 and 5.72 mm). There was a significant correlation of the cartilage volume with the mean thickness (R=0.80) and with the joint surface areas (R=0.56), but not between the thickness and surface area (R=0.37). The association among the patella, tibia, and femur was 0.16 to 0.72 for volumes, 0.08 to 0.78 for thickness, and 0.24 to 0.62 for surfaces. The knee joint cartilage volume and the surface areas were significantly associated with the body height (R=0.51 and 0.57), but not the cartilage thickness (R=0.22). CONCLUSION: There is a surprisingly high variability of the quantitative distribution of cartilage within the knee joint, with only moderate correlations between knee joint cartilage plates, and this variability cannot be adequately predicted based on anthropometric variables.

Adult↗
PubMed2001

The reversed and bidirectional elephant trunk technique in the treatment of complex aortic aneurysms.

BACKGROUND: The elephant trunk technique with a free-floating vascular prosthesis was originally developed to facilitate a subsequent operation on the downstream aorta. We present here our experience with further developments of this technique, which we call the reversed elephant trunk and bidirectional elephant trunk. METHODS: Between January 1, 1995, and December 31, 2000, 505 adult and adolescent patients underwent operations of the thoracic aorta. A reversed elephant trunk procedure in 13 patients and a bidirectional elephant trunk procedure in 4 patients was performed to facilitate either subsequent proximal or proximal and distal aortic replacement. Nine patients underwent subsequent aortic arch replacement with the reversed prosthetic portion after a mean interval of 8 +/- 5.5 months, and 2 patients received distal extension by use of the distal portion of the free-floating graft. RESULTS: There was no hospital mortality (30 days) in this small group of patients, and no patient had aortic rupture, malperfusion caused by the technique itself, or thromboembolic complications during the waiting interval between the first and the second operations. Five patients are still being observed until the contiguous aortic size is large enough to require an operation, and one 74-year-old patient declined a second-stage operation. CONCLUSION: The reversed and bidirectional elephant trunk techniques are interesting options that may be suitable for patients having complex abnormalities of the thoracic aorta and thoracoabdominal aorta when the proximal portion of the descending aorta has to be replaced before the aortic arch with or without the ascending aorta or the distal descending aorta with or without the thoracoabdominal aorta.

Adolescent↗
PubMed2001

Complex host cell responses to antisense suppression of ACHE gene expression.

3'-End-capped, 20-mer antisense oligodeoxynucleotides (AS-ODN) protected with 2'-O-methyl (Me) or phosphorothioate (PS) substitutions were targeted to acetylcholinesterase (AChE) mRNA and studied in PC12 cells. Me-modified AS-ODN suppressed AChE activity up to 50% at concentrations of 0.02-100 nM. PS-ODN was effective at 1-100 nM. Both AS-ODN displayed progressively decreased efficacy above 10 nM. In situ hybridization and confocal microscopy demonstrated dose-dependent decreases, then increases, in AChE mRNA. Moreover, labeling at nuclear foci suggested facilitated transcription or stabilization of AChE mRNA or both under AS-ODN. Intracellular concentrations of biotinylated oligonucleotide equaled those of target mRNA at extracellular concentrations of 0.02 nM yet increased only 6-fold at 1 microM ODN. Above 50 nM, sequence-independent swelling of cellular, but not nuclear, volume was observed. Our findings demonstrate suppressed AChE expression using extremely low concentrations of AS-ODN and attribute reduced efficacy at higher concentrations to complex host cell feedback responses.

Acetylcholinesterase↗
PubMed2000

Multiple conformational states of the hammerhead ribozyme, broad time range of relaxation and topology of dynamics.

The dynamics of a hammerhead ribozyme was analyzed by measurements of fluorescence-detected temperature jump relaxation. The ribozyme was substituted at different positions by 2-aminopurine (2-AP) as fluorescence indicator; these substitutions do not inhibit catalysis. The general shape of relaxation curves reported from different positions of the ribozyme is very similar: a fast decrease of fluorescence, mainly due to physical quenching, is followed by a slower increase of fluorescence due to conformational relaxation. In most cases at least three relaxation time constants in the time range from a few microseconds to approximately 200 ms are required for fitting. Although the relaxation at different positions of the ribozyme is similar in general, suggesting a global type of ribozyme dynamics, a close examination reveals differences, indicating an individual local response. For example, 2-AP in a tetraloop reports mainly the local loop dynamics known from isolated loops, whereas 2-AP located at the core, e.g. at the cleavage site or its vicinity, also reports relatively large amplitudes of slower components of the ribozyme dynamics. A variant with an A-->G substitution in domain II, resulting in an inactive form, leads to the appearance of a particularly slow relaxation process (tau approximately 200 ms). Addition of Mg(2+) ions induces a reduction of amplitudes and in most cases a general increase of time constants. Differences between the hammerhead variants are clearly demonstrated by subtraction of relaxation curves recorded under corresponding conditions. The changes induced in the relaxation response by Mg(2+) are very similar to those induced by Ca(2+). The relaxation data do not provide any evidence for formation of Mg(2+)-inner sphere complexes in hammerhead ribozymes, because a Mg(2+)-specific relaxation effect was not visible. However, a Mg(2+)-specific effect was found for a dodeca-riboadenylate substituted with 2-AP, showing that the fluorescence of 2-AP is able to indicate inner sphere complexation. Amplitudes and time constants show that the equilibrium constant of inner sphere complexation is 1.2, corresponding to 55% inner sphere state of the Mg(2+) complexes; the rate constant 6.6 x 10(3) s(-1) for inner sphere complexation is relatively low and shows the existence of some barrier(s) on the way to inner sphere complexes.

2-Aminopurine↗
PubMed2000

HIV-1 LTR as a target for synthetic ribozyme-mediated inhibition of gene expression: site selection and inhibition in cell culture.

A library of three synthetic ribozymes with randomized arms, targeting NUX, GUX and NXG triplets, respectively, were used to identify ribozyme-accessible sites on the HIV-1 LTR transcript comprising positions -533 to 386. Three cleavable sites were identified at positions 109, 115 and 161. Ribozymes were designed against these sites, either unmodified or with 2'-modifications and phosphorothioate groups, and their cleavage activities of the transcript were determined. Their biological activities were assessed in cell culture, using a HIV-1 model assay system where the LTR is a promoter for the expression of the reporter gene luciferase in a transient expression system. Intracellular efficiency of the ribozymes were determined by cotransfection of ribozyme and plasmid DNA, expressing the target RNA. Modified ribozymes, directed against positions 115 and 161, lowered the level of LTR mRNA in the cell resulting in inhibition of expression of the LTR-driven reporter gene luciferase of 87 and 61%, respectively. In the presence of Tat the inhibitions were 43 and 25%. The inactive variants of these ribozymes exhibited a similar inhibitory effect. RNase protection revealed a reduction of RNA which was somewhat stronger for the active than the inactive ribozymes, particularly for ribozyme 115. Unmodified ribozymes showed no inhibition in the cell. The third ribozyme, targeting a GUG-triplet at position 109, possessed only low cleavage activity in vitro and no inhibitory effect in cell culture.

Base Sequence↗
PubMed2000

In vitro evolution of the hammerhead ribozyme to a purine-specific ribozyme using mutagenic PCR with two nucleotide analogues.

The conventional hammerhead ribozyme cleaves RNA 3' to nucleotide triplets with the general formula NUH, where N is any nucleotide, U is uridine and H is any nucleotide except guanosine. In order to isolate hammerhead ribozyme sequences capable of cleaving 3' to the GUG triplet, we performed a mutagenic selection protocol starting with the conventional sequence of an NUH-cleaving ribozyme. The 22 nucleotides in the core and the stem-loop II region were subjected to mutagenic PCR using the two nucleotide analogues 6-(2-deoxy-beta-d-ribofuranosyl)-3,4-dihydro-8H-pyrimido-[4,5-C)][1, 2] oxazin-7-one and of 8-oxo-2'-deoxyguanosine. After five repetitions of the selection cycle, several clones showed cleavage activity. One sequence, having one deletion, showed at least a 90 times higher in trans cleavage rate than the starting ribozyme. It cleaved 3' to GUG and GUA. The sequence of this ribozyme is essentially identical with that obtained previously by selection for AUG cleavage starting with a randomised core and stem-loop II region. This identical result of two independent selection procedures supports the notion that sequences for NUR cleavage, where R is a purine nucleotide, are not compatible with the classical hammerhead structure, and that the sequence space for this cleavage specificity is very limited. The cleavage of NUR triplets is not restricted to the sequence of the substrate that was used for selection but is sequence-independent for in trans cleavage, although the sequence context influences the value for the cleavage rate somewhat. Analysis of cleavage activities indicates the importance of A at position L2.5 in loop II.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed2000

Lipid phosphorylation in chloroplast envelopes. Evidence for galactolipid CTP-dependent kinase activities.

Lipid phosphorylation takes place within the chloroplast envelope. In addition to phosphatidic acid, phosphatidylinositol phosphate, and their corresponding lyso-derivatives, we found that two novel lipids underwent phosphorylation in envelopes, particularly in the presence of carrier-free [gamma-(32)P]ATP. These two lipids incorporated radioactive phosphate in chloroplasts in the presence of [gamma-(32)P]ATP or [(32)P]P(i) and light. Interestingly, these two lipids were preferentially phosphorylated in envelope membranes in the presence [gamma-(32)P]CTP, as the phosphoryl donor, or [gamma-(32)P]ATP, when supplemented with CDP and nucleoside diphosphate kinase II. The lipid kinase activity involved in this reaction was specifically inhibited in the presence of cytosine 5'-O-(thiotriphosphate) (CTPgammaS) and sensitive to CTP chase, thereby showing that both lipids are phosphorylated by an envelope CTP-dependent lipid kinase. The lipids were identified as phosphorylated galactolipids by using an acid hydrolysis procedure that generated galactose 6-phosphate. CTPgammaS did not affect the import of the small ribulose-bisphosphate carboxylase/oxygenase subunit into chloroplasts, the possible physiological role of this novel CTP-dependent galactolipid kinase activity in the chloroplast envelope is discussed.

Chloroplasts↗
PubMed2000

Dynamics of the RNA hairpin GNRA tetraloop.

The dynamics of RNA hairpin tetraloops of the GNRA type [sequence G- any ribonucleotide (N)-purine (R)-A] was analyzed by fluorescence spectroscopy and by fluorescence-detected temperature-jump relaxation, using RNA oligomers with 2-aminopurine (2AP) substituted in two different positions of the loop sequence, Gp2APpApA (HP1) and GpAp2APpA (HP2), as indicator. The fluorescence of HP1 is much higher than that of HP2, indicating a lower degree of 2AP-stacking in HP1. Addition of Mg(2+) or Ca(2+) ions leads to an increase of fluorescence in HP1, whereas a decrease of fluorescence is observed in HP2. In both cases at least two ion-binding equilibria are required to fit titration data. T-jump experiments using fluorescence detection show a relaxation process with a time constant of 22 micros for HP1, whereas two relaxation processes with time constants 5 and 41 micros, are found for HP2. These results clearly demonstrate the existence of more than the single conformation state detected by NMR analysis. The T-jump amplitudes decrease with increasing bivalent ion concentration, indicating that one of the states is favored in the presence of bivalent ions. The loop relaxation processes are slower than standard stacking processes, probably because of activation barriers imposed by a restricted mobility of loop residues, and are assigned to a stacking rearrangement, probably between the 5' and the 3'-side. A similar process has been observed previously for the anticodon loop of tRNA(Phe). The rate constants of the transition are in the range of 10(4) s(-1) in the case of HP1. The data demonstrate the existence of structures that are not resolved by standard NMR because of fast exchange and are not found by X-ray analysis because of restrictions by crystal packing.

2-Aminopurine↗
PubMed2000

Effect of gradient and section orientation on quantitative analysis of knee joint cartilage.

The object of this study was to determine the influence of the gradient and section orientation on cartilage thickness and volume measurements in the knee joint. Eight specimens were imaged with a fat-suppressed gradient-echo sequence, applying sagittal, transverse, and coronal section orientations. Images were additionally acquired with exchanged gradient directions, and with computed tomography (CT) arthrography. After segmentation and three-dimensional (3D) reconstruction, the volume, the mean, and the maximal 3D cartilage thickness were computed. No effect of changes in the gradient orientation was found, suggesting that susceptibility-induced geometric distortion is not a relevant problem in quantitative cartilage imaging. Sagittal images produced similar data to that obtained with transverse (patella) or coronal (tibia) sections, demonstrating that all knee joint cartilages can be accurately quantified from a single sagittal data set. Whereas no significant systematic deviation between magnetic resonance imaging (MRI) and CT arthrography was recorded in the patella, there was a 10%-15% underestimation of tibial cartilage thickness in MRI.

Cartilage, Articular↗
PubMed2000

Elastic registration of 3D cartilage surfaces from MR image data for detecting local changes in cartilage thickness.

The objective of this work was to develop and validate a computational method for the registration (matching) of 3D cartilage plates from MR image data sets. The technique tracks local cartilage thickness changes over time. A 3D elastic registration technique was applied that identifies corresponding points of the bone-cartilage interface in MR data sets of 3D-reconstructed cartilage plates. In a first rigid preregistration step, the surfaces are aligned, using the principal axes decomposition to correct for different joint positions and orientations in the MR scanner. In a second step, the surfaces are deformed elastically, based on geometric surface features, until they are sufficiently similar to identify corresponding surface points. The method was validated against artificially corrupted cartilage surfaces and MR data obtained from in vivo and in vitro compression experiments. The in vivo reproducibility was tested on patellar data sets of volunteers, with repositioning of the joint in between replicate acquisitions.

Cartilage, Articular↗
PubMed2000

In situ femoral dual-energy X-ray absorptiometry related to ash weight, bone size and density, and its relationship with mechanical failure loads of the proximal femur.

The objective of this study was to directly compare in situ femoral dual-energy X-ray absorptiometry (DXA) and in vitro chemical analysis (ash weight and calcium) with mechanical failure loads of the proximal femur, and to determine the influence of bone size (volume) and density on mechanical failure and DXA-derived areal bone mineral density (BMD, in g/cm2). We performed femoral DXA in 52 fixed cadavers (age 82.1 +/- 9.7 years; 30 male, 22 female) with intact skin and soft tissues. The femora were then excised, mechanically loaded to failure in a stance phase configuration, their volume measured with a water displacement method (proximal neck to lesser trochanter), and the ash weight and calcium content of this region determined by chemical analysis. The correlation coefficient between the bone mineral content (measured in situ with DXA) and the ash weight was r = 0.87 (standard error of the estimate = 16%), the ash weight allowing for a better prediction of femoral failure loads (r = 0.78; p < 0.01) than DXA (r = 0.67; p < 0.01). The femoral volume (r = 0.61; p < 0.01), but not the volumetric bone density (r = 0.26), was significantly associated with the failure load. The femoral bone volume had a significant impact (r = 0.35; p < 0.01) on the areal BMD (DXA), and only 63% of the variability of bone volume could be predicted (based on the basis of body height, weight and femoral projectional bone area. The results suggest that accuracy errors of femoral DXA limit the prediction of mechanical failure loads, and that the influence of bone size on areal BMD cannot be fully corrected by accounting for body height, weight and projected femoral area.

Absorptiometry, Photon↗
PubMed2000

Precision of tibial cartilage morphometry with a coronal water-excitation MR sequence.

The aim of this study was to analyze the precision of tibial cartilage morphometry, by using a fast, coronal water-excitation sequence with high spatial resolution, to compare the reproducibility of 3D thickness vs volume estimates, and to test the technique in patients with severe osteoarthritis. The tibiae of 8 healthy volunteers and 3 patients selected for total knee arthroplasty were imaged repeatedly with a water-excitation sequence (image time 6 h 19 min, resolution 1.2 x 0.31 x 0.31 mm(3)), with the knee being repositioned between each replicate acquisition. After 3D reconstruction, the cartilage volume, the mean, and the maximal tibial cartilage thickness were determined by 3D Euclidean distance transformation. In the volunteers, the precision of the volume measurements was 2.3 % (CV%) in the medial and 2.6 % in the lateral tibia. The reproducibility of the mean cartilage thickness was similar (2.6 and 2.5 %, respectively), and that of the maximal thickness lower (6.5 and 4.4 %). The patients showed a considerable reduction in volume and thickness, the precision being comparable with that in the volunteers. We find that, using a new imaging protocol and computational algorithm, it is possible to determine tibial cartilage morphometry with high precision in healthy individuals as well as in patients with osteoarthritis.

Adult↗
PubMed2000

Patellar cartilage deformation in vivo after static versus dynamic loading.

The objective of this study was to test the hypothesis that static loading (squatting at a 90 degrees angle) and dynamic loading (30 deep knee bends) cause different extents and patterns of patellar cartilage deformation in vivo. The two activities were selected because they imply different types of joint loading and reflect a realistic and appropriate range of strenuous activity. Twelve healthy volunteers were examined and the volume and thickness of the patellar cartilage determined before and from 90 to 320s after loading, using a water excitation gradient echo MR sequence and a three-dimensional (3D) distance transformation algorithm. Following knee bends, we observed a residual reduction of the patellar cartilage volume (-5.9+/-2.1%; p<0.01) and of the maximal cartilage thickness (-2.8+/-2.6%), the maximal deformation occurring in the superior lateral and the medial patellar facet. Following squatting, the change of patellar cartilage volume was -4.7+/-1.6% (p<0.01) and that of the maximal cartilage thickness -4.9+/-1.4% (p<0.01), the maximal deformation being recorded in the central aspect of the lateral patellar facet. The volume changes were significantly lower after squatting than after knee bends (p<0.05), but the maximal thickness changes higher (p<0.05). The results obtained in this study can serve to validate computer models of joint load transfer, to guide experiments on the mechanical regulation of chondrocyte biosynthesis, and to estimate the magnitude of deformation to be encountered by tissue-engineered cartilage within its target environment.

Adult↗