Biomedical subjects

F Eckstein

Publications and source records attributed to F Eckstein.

At least 109 records · Page 6Linked to original sources

PubMed1999

Determination of calcaneal ultrasound properties ex situ: reproducibility, effects of storage, formalin fixation, maceration, and changes in anatomic measurement site.

The objective of this study was to determine the reproducibility of ultrasonic bone properties with a system for measuring calcanei ex situ; the influence of changes of the measurement site; and the effects of fixation, storage, and maceration. We examined 14 fixed calcanei and 12 fresh bones. Ultrasonic measurements were performed ex situ after degassing, using an Achilles+ system and a special positioning device. The instrument precision was 0.16% for speed of sound (SOS), 1.4% for broadband ultrasonic attenuation (BUA), and 1. 8% for the stiffness index (SI). The short-term precision was 0.54%, 1.9%, and 2.8%, respectively. A defined shift of the measurement site (5 mm distal of the middle) led to unpredictable changes in ultrasound (US) properties (r = 0.65 for SOS, 0.82 for BUA, and 0.75 for SI). Embalment with 4% formalin/96% alcohol caused a systematic decrease in SOS, an increase in BUA, and a decrease in SI (mean = -12.7 units; P < 0.001), the effect increasing with time. However, values at 6 months of fixation and later were highly correlated with those in fresh specimens (r = 0.95 for the SI). Two weeks storage in degassed and normal solution had only modest effects on ultrasound properties. Maceration did not lead to a systematic increase or decrease of ultrasound variables, but introduced unpredictable changes (r = 0.64-0.94). We conclude that in comparative biomechanical studies it is feasible to measure calcaneal specimens embalmed in formalin/alcohol ex situ, if the primary interest is not in the absolute values but in the correlation with mechanical failure loads at other skeletal sites.

Aged↗

PubMed1999

Precision and intersite correlation of bone densitometry at the radius, tibia and femur with peripheral quantitative CT.

OBJECTIVE: To compare the in situ precision of peripheral quantitative CT (pQCT) at the radius, tibia and femur, and to analyze the intersite correlation, in order to determine whether measurements at the lower extremity reproduce results at the radius or are of additional informative value. DESIGN AND MATERIAL: pQCT measurements were performed in 86 elderly cadavers (mean age 80.5 years) at trabecular and cortical locations in the radius, tibia and femur, determining densitometric (bone mineral content and density) as well as geometric parameters (cross-sectional area, cortical thickness, polar moment of inertia and others). In 14 cadavers, repeated measurements were obtained at all sites on four different days. RESULTS AND CONCLUSIONS: At cortical sites, the precision for the densitometric and geometric variables ranged from 0.4% to 4.3%, and was similar for the radius, tibia and femur. At trabecular locations, the reproducibility of the density measurements ranged from 1.8% to 2.5% at the radius, and from 3.2% to 5.9% at the femur and tibia. The intersite correlation of the total bone mineral content ranged from 0.87 and 0.97 at cortical sites, and from 0.63 to 0.85 at trabecular locations. The trabecular density showed a higher similarity between the tibia and femur (r=0.68-0.78) than between the radius and the lower extremity (r=0.41-0.45). The results demonstrate a substantial heterogeneity of trabecular bone in elderly individuals and advocate measurements directly at the site of clinical or scientific interest.

Absorptiometry, Photon↗

PubMed1999

Tension and bending, but not compression alone determine the functional adaptation of subchondral bone in incongruous joints.

In the present study, we tested the hypothesis that tension and bending, rather than compression alone, determine the functional adaptation of subchondral bone in incongruous joints. We investigated whether tensile stresses in the subchondral bone of the humero-ulnar articulation are affected by the direction of muscle and joint forces, and whether the tensile stresses are large enough to cause microstructural adaptation, specifically a preferential alignment of the trabeculae and the subchondral collagen fibres. Using a previously validated finite element model of the human humero-ulnar joint, we calculated the contact pressure, the principal compressive and tensile stresses, and the strain energy density in the subchondral bone for various flexion angles. A bicentric (ventro-dorsal) pressure distribution was found in the joint at 30 degrees to 120 degrees of flexion, with contact pressures of up to between 2.5 and 3 MPa in the ventral and dorsal aspects of the ulnar joint surface, but less than 0.5 MPa in the centre. The principal tensile stress in the subchondral bone of the trochlear notch quantitatively exceeded the principal compressive stress at low flexion angles (maximum 8.2 MPa), and the distribution of subchondral strain energy density differed substantially from that of the contact stress (r=-0.72 at 30 degrees and r=+0.58 at 90 degrees of flexion). No important tensile stress was computed in the trochlea humeri. On contact radiography, we found sagittally orientated subarticular trabeculae in the notch, running tangential to the surface. Furthermore, we observed sagittally orientated split lines in the subchondral bone of the notch of 20 cadaver joints, suggesting a ventro-dorsal orientation of the collagen fibres. The trochlea humeri, on the other hand, did not show a preferential direction of the subchondral split lines, these findings confirming the predictions of tensile stresses in the model. We conclude that, due to the important contribution of tension to subchondral bone stress, the distribution of subchondral density cannot be directly employed for assessing the long term distribution of joint pressure at the cartilage surface. The magnitude of the tensional stress varies considerably with the direction of the muscle and joint forces, and it appears large enough to cause functional adaptation of the subchondral bone on a microstructural level.

Adaptation, Physiological↗

PubMed1999

A non-destructive technique for 3-D microstructural phenotypic characterisation of bones in genetically altered mice: preliminary data in growth hormone transgenic animals and normal controls.

A non-destructive, three-dimensional technique for microstructural phenotypic characterisation of skeletal elements in genetically altered mice is presented. Preliminary data in bovine growth-hormone transgenic animals and control littermates are shown. The technique is based on microcomputed tomography (microCT) and digital postprocessing and allows for a differential quantitative analysis of the cortical and trabecular bone compartments in the axial and peripheral skeleton. The distal femora and the first lumbar vertebral bodies of six animals were CT scanned in the axial plane with an isotropic resolution of 20 microm. The periostal surface and the marrow spaces were segmented fully automatically, and the trabecular and cortical compartments were separated interactively. After 3-D reconstruction, various regions of interest (diaphyseal, metaphyseal and epiphyseal) were selected for the analysis. The femora and vertebrae of the transgenic animals showed obvious differences in size, shape, and trabecular arrangement compared with the control animals. The total bone mass was increased by a factor of two to three, but the trabecular bone was increased much more (up to 12 times) than the cortical bone. The transgenic animals showed an increased ratio of trabecular vs cortical bone (0.90 to 1.27 vs 0.14 to 0.36 in the femoral diaphysis) and an elevated trabecular bone volume fraction (49% to 73% vs 18% to 43% in the femoral metaphysis). The mean 3-D cortical thickness was similar in the normal and transgenic animals (values between 93 microm and 232 microm in the dia- and metaphyses), but the minimal cortical thickness was lower in the transgenic animals (22 to 31 microm vs 54 microm to 110 microm in the diaphysis). The technique presented is suitable for phenotypic characterisation of bone structure in genetically altered mice.

Animals↗

PubMed1999

Functional analysis of articular cartilage deformation, recovery, and fluid flow following dynamic exercise in vivo.

The function of articular cartilage depends on the interaction between the tissue matrix and the interstitial fluid bound to the proteoglycan molecules. Mechanical loading has been shown to be involved in both the metabolic regulation of chondrocytes and in matrix degeneration. The purpose of the present study was therefore to analyze the deformation, recovery, and fluid flow in human articular cartilage after dynamic loading in vivo. The patellae of 7 volunteers were imaged at physical rest and after performing knee bends, with a specifically optimized fat-suppressed FLASH-3D magnetic resonance (MR) sequence. To measure cartilage deformation, the total volume of the patellar cartilage was determined, employing 3D digital image analysis. Patellar cartilage deformation ranged from 2.4 to 8.6% after 50 knee bends, and from 2.4% to 8.5% after 100 knee bends. Repeated sets of dynamic exercise at intervals of 15 min did not cause further deformation. After 100 knee bends, the cartilage required more than 90 min to recover from loading. The rate of fluid flow during relaxation ranged from 1.1 to 3.5 mm(3)/min (0.08 to 0.22 mm(3)/min per square centimeter of the articular surface) and was highly correlated with the individual degree of deformation after knee bends. The data provide the first quantification of articular cartilage recovery and of the rate of fluid flow between the cartilage matrix and surrounding tissue in intact joints in vivo. Measurement in the living opens the possibility of relating interindividual variations of mechanical cartilage properties to the susceptibility of developing joint failure, to assess the load-partitioning between the fluid phase and solid cartilage matrix during load transfer, and to determine the role of mechanically induced fluid flow in the regulation of the metabolic activity of chondrocytes.

Adult↗

PubMed1999

In situ measurement of articular cartilage deformation in intact femoropatellar joints under static loading.

The deformational behavior of articular cartilage has been investigated in confined and unconfined compression experiments and indentation tests, but to date there exist no reliable data on the in situ deformation of the cartilage during static loading. The objective of the current study was to perform a systematic study into cartilage compression of intact human femoro-patellar joints under short- and long-term static loading with MR imaging. A non-metallic pneumatic pressure device was used to apply loads of 150% body weight to six joints within the extremity coil of an MRI scanner. The cartilage was delineated during the compression experiment with previously validated 2D and 3D fat-suppressed gradient echo sequences. We observed a mean (maximal) in situ deformation of 44% (57%) in patellar cartilage after 32 h of loading (mean contact pressure 3.6 MPa), the femoral cartilage showing a smaller amount of deformation than the patella. However, only around 7% of the final deformation (3% absolute deformation) occurred during the first minute of loading. A 43% fluid loss from the interstitial patellar matrix was recorded, the initial fluid flux being 0.217 +/- 0.083 microm/s, and a high inter-individual variability of the deformational behavior (coefficients of variation 11-38%). In conjunction with finite-element analyses, these data may be used to compute the load partitioning between the solid matrix and fluid phase, and to elucidate the etiologic factors relevant in mechanically induced osteoarthritis. They can also provide direct estimates of the mechanical strain to be encountered by cartilage transplants.

Adolescent↗

PubMed1999

Interobserver reproducibility of quantitative cartilage measurements: comparison of B-spline snakes and manual segmentation.

The objective of this work was to develop a segmentation technique for thickness measurements of the articular cartilage in MR images and to assess the interobserver reproducibility of the method in comparison with manual segmentation. The algorithm is based on a B-spline snakes approach and is able to delineate the cartilage boundaries in real time and with minimal user interaction. The interobserver reproducibility of the method, ranging from 3.3 to 13.6% for various section orientations and joint surfaces, proved to be significantly superior to manual segmentation.

Cartilage, Articular↗

PubMed1999

[Preferential direction of collagen fibrils in the sub-chondral bone bone and the hip and shoulder joint].

We hypothesised that--due to bending and tension--there should exist a preferential direction of the collagen fibrils in the subchondral bone of the concave components of the hip and shoulder joint that results from bicentric or eccentric loading, but there should be no preferential direction in the convex joint partners. We therefore examined 25 human hip and 27 shoulder joints, these being decalcified after maceration. To analyse the preferential direction of the collagen fibrils, we used the split line method. The subchondral plate was pierced at regular intervals with needles that had been previously dipped in diluted ink. In the acetabulum, we found a predominantly transverse direction of the split lines in the ventral and dorsal horn of the lunatic surface, and these usually continued through the acetabular fossa. In the ventral part of the acetabular roof, arch-like orientations of the split lines were observed. In the glenoid cavity, a clearly preferential orientation was found in anterior-posterior direction, usually in the middle third of the articular surface. In the femoral and humeral heads, no preferential direction of the split lines was observed in any of the specimens. We interpret the split-line patterns in the acetabulum as an expression of the tensile stress that is encountered during a "spreading open" of the socket upon bicentric (ventral-dorsal) loading in the physiologically incongruous joint. In the glenoid, the relatively weak bony support in the ventral and dorsal part of the articular surface may be responsible for bending and tensile stress, particularly in view of eccentric loading during dynamic activity, and this could explain the observed anterio-posterior split line pattern. The results support the idea that the subchondral bone of concave joint partners encounters tension, leading to a preferential direction of the collagen fibrils. This can be considered as a functional adaptation of the subchondral bone on a microstructural level.

Aged↗

PubMed1999

[Sex-specific analysis of bone mass in normal and growth hormone transgenic mice using dual energy x-ray absorptiometry (DXA)].

The aim of the present study was the non-invasive, sex-specific measurement of bone mass in bovine growth hormone (bGH) transgenic mice and normal controls with dual energy X-ray absorptiometry (DXA). The transgenic mouse constitutes a suitable animal model to study the influence of growth hormone on the skeletal system. We analysed 28 animals, aged 12 weeks (14 transgenic, 14 controls, 7 male and 7 female, respectively), using a peripheral DXA scanner that had been adapted to the measurement of small animals. At a measurement time of 20 min, the precision (RMS average CV%) was 4.4% for bone mass (BMC), 2.5% for areal bone density (BMD), 0.86% for total body weight and 4.5% for the percentage BMC (relative to body weight). While the absolute bone mass was not significantly different between male and female animals, we found a higher percentage of the BMC relative to the total bone mass in females (+21% in controls, +31% in transgenics; p < 0.01). The absolute bone mass was higher in the transgenic animals (+71% in females and +62% in males; p < 0.01), but relative to the body weight the transgenic females yielded similar and the transgenic males lower values (-7.2%; p < 0.05). Using DXA it is possible to non-invasively determine the mass of mineralised tissue in the mouse with relatively high precision and to effectively discriminate between different groups. Although a strong influence of growth hormone on the absolute bone mass is observed, the results show that this increase is not higher than that of the total body mass.

Absorptiometry, Photon↗

PubMed1999

Nucleic acid enzymes.

Last year provided new structural data, particularly for the group I intron and the Hepatitis delta ribozymes, that were essential for a better understanding of the RNA structure/function relationship. The role of metal ions in catalysis of ribozyme action still remains elusive, however. In vitro selection has continued to be a rich source for obtaining data on new nucleic acid enzyme activities.

Base Sequence↗

PubMed1999

Comparison of quantitative ultrasound in the human calcaneus with mechanical failure loads of the hip and spine.

OBJECTIVE: Quantitative ultrasound of the calcaneus is used clinically for evaluating bone fracture risk, but its association with the mechanical properties at other skeletal sites is not well characterized. The objective was therefore to determine its predictive ability of the mechanical failure loads of the proximal femur and lumbar spine. METHOD: In 45 human cadavers (29 males and 16 females, aged 82.5 +/- 9.6 years), we determined the speed of sound, broadband ultrasonic attenuation (BUA) and the empirical stiffness index, using a commercial quantitative ultrasound scanner. The proximal femora and the fourth vertebral body were excised and loaded to failure in a testing machine. RESULTS: Femoral failure loads ranged from 933 to 7000 N and those of the vertebrae from 1000 to 7867 N, their correlation being 0.51 in females and -0.08 in males. Forty percent of the variability of femoral, but only 24% of the variability of the vertebral fracture loads could be predicted with calcaneal speed of sound. In the femur, a combination of speed of sound and BUA improved the prediction (r2 = 50-60%), but not in the spine. CONCLUSIONS: The study provides experimental evidence that calcaneal quantitative ultrasound is capable of predicting mechanical failure at other skeletal sites and has potential to identify patients at risk from osteoporotic fracture. The different association of quantitative ultrasound with femoral and vertebral failure may result from the influence of the cortical bone and a higher microstructure-related similarity of the calcaneus and the femur.

Aged↗

PubMed1999

Transient transfection of a synthetic hammerhead ribozyme targeted against human MGMT gene to cells in culture potentiates the genotoxicity of the alkylation damage induced by mitozolomide.

Unmodified and chemically modified forms of a synthetic hammerhead ribozyme with the mRNA of methylguanine-DNA methyltransferase (MGMT) gene as substrate were characterized for their in vitro and in vivo activities. The unmodified ribozyme efficiently cleaved in vitro a short synthetic substrate, and it was rapidly degraded in fetal bovine serum (FBS). The introduction of phosphorothioates and the substitution of uridine with thymidine at probable nuclease-sensitive sites slightly increased the nuclease resistance of the ribozyme. Conversely, pyrimidine nucleoside substitution with 2'NH2 and 2'F nucleosides strongly enhanced nuclease resistance. The in vivo activity was determined by measuring the genotoxicity induced by the alkylating drug mitozolomide, the damage of which is repaired by MGMT enzyme. CHO/47 cells, temporarily depleted of the MGMT protein, were first transfected with the various synthetic ribozymes and subsequently treated with mitozolomide. At equivalent concentration of the drug, the induction of sister chromatid exchanges was higher in ribozyme-transfected than in untransfected cells, indicating that the synthetic ribozymes potentiated the genotoxicity of mitozolomide. Moreover, the concomitant occurrence of messenger RNA reduction in ribozyme-transfected cells indicated that the inhibition of MGMT resynthesis was the basis of the enhanced genotoxicity.

Alkylating Agents↗

PubMed1999

Antisense technologies have a future fighting neurodegenerative diseases.

Our growing understanding of the role that unfavorable patterns of gene expression play in the etiology of neurodegenerative disease emphasizes the need for strategies to selectively block the biosynthesis of harmful proteins in the brain. Antisense technologies are ideally suited to this purpose. Tailor-designed to target specific RNA, antisense oligonucleotides and ribozymes offer tools to suppress the production of proteins mediating neurodegeneration. Although technical limitations must still be overcome, the antisense approach represents a novel and exciting strategy for intervention in diseases of the central nervous system.

Animals↗

PubMed1999

Human osteogenesis involves differentiation-dependent increases in the morphogenically active 3' alternative splicing variant of acetylcholinesterase.

The extended human acetylcholinesterase (AChE) promoter contains many binding sites for osteogenic factors, including 1,25-(OH)2 vitamin D3 and 17beta-estradiol. In differentiating osteosarcoma Saos-2 cells, both of these factors enhanced transcription of the AChE mRNA variant 3' terminated with exon 6 (E6-AChE mRNA), which encodes the catalytically and morphogenically active E6-AChE isoform. In contrast, antisense oligodeoxynucleotide suppression of E6-AChE mRNA expression increased Saos-2 proliferation in a dose- and sequence-dependent manner. The antisense mechanism of action was most likely mediated by mRNA destruction or translational arrest, as cytochemical staining revealed reduction in AChE gene expression. In vivo, we found that E6-AChE mRNA levels rose following midgestation in normally differentiating, postproliferative fetal chondrocytes but not in the osteogenically impaired chondrocytes of dwarf fetuses with thanatophoric dysplasia. Taken together, these findings suggest morphogenic involvement of E6-AChE in the proliferation-differentiation balance characteristic of human osteogenesis.

Acetylcholinesterase↗

PubMed1999

Three-dimensional analysis of the width of the subacromial space in healthy subjects and patients with impingement syndrome.

OBJECTIVE: The aim of this study was to per form a three-dimensional analysis of the width of the subacromial space during passive and active arm abduction in healthy volunteers and patients with impingement syndrome. SUBJECTS AND METHODS: The shoulders of 10 healthy subjects and 10 patients with impingement syndrome were imaged with an open MR system during abduction, with and without activation of the shoulder muscles. An apparatus was designed for applying an adduction force of 10 N to the distal humerus during image acquisition, and the minimal acromiohumeral distance was measured after three-dimensional reconstruction. RESULTS: In the 10 healthy volunteers, muscle activity led to a significant decrease (-32%; p < .05) of the acromiohumeral distance at 60 degrees of abduction, whereas at 120 degrees of abduction the distance was significantly increased (+44%; p < .05). In these volunteers, muscle activation caused no significant effect at 90 degrees of abduction. However, in the 10 patients with impingement syndrome, muscle activity led to a significant decrease in the width of the subacromial space compared with that of the healthy contralateral side (-68%; p < .05). CONCLUSION: Muscle activity and arm position were found to cause systematic changes in the width of the subacromial space. However, functional deficits of the supraspinous muscle in patients with early-stage impingement syndrome were not apparent during muscle relaxation.

Acromion↗

PubMed1998

Recent developments in the hammerhead ribozyme field.

Developments in the hammerhead ribozyme field during the last two years are reviewed here. New results on the specificity of this ribozyme, the mechanism of its action and on the question of metal ion involvement in the cleavage reaction are discussed. To demonstrate the potential of ribozyme technology examples of the application of this ribozyme for the inhibition of gene expression in cell culture, in animals, as well as in plant models are presented. Particular emphasis is given to critical steps in the approach, including RNA site selection, delivery, vector development and cassette construction.

Animals↗

PubMed1998

Incorporation of terminal phosphorothioates into oligonucleotides.

Considerable effort has been directed towards studying the structure and function of oligonucleotides and several approaches rely on the attachment of reporter groups to oligonucleotides. We report here the introduction of 3'- and 5'-terminal phosphorothioates into heptameric oligonucleotides and their post-synthetic modification with several reporter groups. The synthesis of terminal phosphorothioates is based on the coupling of a ribonucleoside phosphoramidite at the first or last nucleotide, respectively, which, after sulphurization, is removed by sequential oxidation of the vicinal hydroxyl groups and then beta-elimination. Product formation is of the order of 95%. The ratio of phosphorothioate- versus phosphate-terminated oligodeoxynucleotides as analysed by electrophoresis on a Hg2+gel is in general 85/15. Examples for the reactivity of the terminal phosphorothioates for conjugation with cholesterol, bimane and for sulphydryl exchange are described.

Bridged Bicyclo Compounds↗

PubMed1998

Sequence specificity of the hammerhead ribozyme revisited; the NHH rule.

The sequence specificity of hammerhead ribozyme cleavage has been re-evaluated with respect to the NUH rule. Contrary to previous reports it was found that substrates with GAC triplets were also cleaved. This was established in three different sequence contexts. The rate of cleavage under single turnover conditions was between 3 and 7% that of cleavage 3' of GUC. Specificity of cleavage of substrates containing a central A in the cleavable triplet can be described as NAH, where N can be any nucleotide and H any nucleotide but G. As cleavage 3' of NCH triplets has recently been described, the NUH rule can be reformulated to NHH.

Base Sequence↗