A method for measuring joint kinematics designed for accurate registration of kinematic data to models constructed from CT data.

A method for measuring three-dimensional kinematics that incorporates the direct cross-registration of experimental kinematics with anatomic geometry from Computed Tomography (CT) data has been developed. Plexiglas registration blocks were attached to the bones of interest and the specimen was CT scanned. Computer models of the bone surface were developed from the CT image data. Determination of discrete kinematics was accomplished by digitizing three pre-selected contiguous surfaces of each registration block using a three-dimensional point digitization system. Cross-registration of bone surface models from the CT data was accomplished by identifying the registration block surfaces within the CT images. Kinematics measured during a biomechanical experiment were applied to the computer models of the bone surface. The overall accuracy of the method was shown to be at or below the accuracy of the digitization system used. For this experimental application, the accuracy was better than +/-0.1mm for position and 0.1 degrees for orientation for linkage digitization and better than +/-0.2mm and +/-0.2 degrees for CT digitization. Surface models of the radius and ulna were constructed from CT data, as an example application. Kinematics of the bones were measured for simulated forearm rotation. Screw-displacement axis analysis showed 0.1mm (proximal) translation of the radius (with respect to the ulna) from supination to neutral (85.2 degrees rotation) and 1.4mm (proximal) translation from neutral to pronation (65.3 degrees rotation). The motion of the radius with respect to the ulna was displayed using the surface models. This methodology is a useful tool for the measurement and application of rigid-body kinematics to computer models.

Forearm rotation alters interosseous ligament strain distribution.

Recent interest in reconstruction of the interosseous ligament (IOL) of the forearm has led to questions concerning optimal placement of the reconstructive graft as well as the ideal rotational position of the forearm during graft tensioning. We therefore studied the strain distribution in the IOL to determine which fibers are strained in different positions of forearm rotation. Five cadaveric human forearms were subjected to compressive axial load (simulating power grip) and the strain values across the entire IOL were measured with the forearm in neutral, supination, and pronation. The strain distribution in the IOL changed with forearm rotation. The highest overall strain was found in neutral. In neutral and pronation, higher strain was observed in the proximal region of the IOL. In supination, however, higher average strain was seen in the distal region of the IOL. These results suggest that a reconstructive graft placed in the proximal region of the IOL and tensioned in neutral rotation would provide balanced constraint in different positions of forearm rotation. A graft placed in the distal region and tensioned in forearm neutral, however, may limit forearm rotation.

Role of the forearm interosseous ligament: is it more than just longitudinal load transfer?

The objective of our study was to measure 3-dimensional force vectors (magnitude and direction) acting in the forearm when load is applied to the hand and to measure the actual force in the interosseous ligament (IOL). Fourteen cadaveric forearms were loaded to 136 N of compression while special load cells measured force vectors in the forearm. Computer forearm models were used to display the 3-dimensional force vector directions. The study results showed that the radius bears most of the load at the wrist but load on the radius at the elbow is reduced because the IOL transfers load to the ulna between the wrist and the elbow. In addition to this role in longitudinal load transfer, our measurement of 3-dimensional forces allowed identification of transverse vectors which suggest that the IOL also functions to keep the radius and ulna from splaying apart. Our results imply that the IOL participates not only in longitudinal load transfer but also in the maintenance of transverse stability of the forearm during compressive load transfer from the hand to the elbow.

Feasibility of partial A2 and A4 pulley excision: residual pulley strength.

We investigated residual digital flexor pulley strengths after 75% excision of the A2 and A4 pulleys. For direct pull-off tests, A2 and A4 pulleys from cadaveric fingers were tested by pulling on a loop of flexor digitorum profundus tendon through the pulley. For functional loading tests, fingers were positioned with the metacarpophalangeal joint flexed to 90 degrees for A2 testing, and with the proximal interphalangeal joint in 90 degrees flexion for A4 testing (with all other joints in full extension). Excision of 75% of A2 and A4 pulleys reduced pulley strengths determined by both testing methods. For the functional loading tests, which are more clinically relevant, mean tendon forces at failure after partial excision of A2 and A4 pulleys were 224 and 131 N respectively, which is sufficient to withstand flexor tendon forces expected during activities of daily living.

A new methodology to measure load transfer through the forearm using multiple universal force sensors.

Previous approaches to measuring forces in the forearm have made the assumption that forces acting in the radius and ulna are uniaxial near the wrist and elbow. To accurately describe forces in the forearm and the forces in the interosseous ligament, we have developed a new methodology to quantitatively determine the 3-D force vectors acting in forearm structures when a compressive load is applied to the hand. A materials testing machine equipped with a six degree-of-freedom universal force-moment sensor (UFS) was employed to apply a uniaxial compressive force to cadaveric forearms gripped at the hand and humerus. Miniature UFSs were implanted into the distal radius and proximal ulna to measure force vectors there. A 3-D digitizing device was used to measure transformations between UFS coordinate systems, utilized for calculating the force vectors in the distal ulna, proximal radius, and the interosseous ligament (IOL). This method was found to be repeatable to within 3 N, and accurate to within 2 N for force magnitudes. Computer models of the forearm, generated from CT scans, were used to visualize the force vectors in 3-D. Application of this methodology to eight forearm specimens showed that the radius carries most of the load at the wrist while force in the IOL relieves load acting in the radius at the mid-forearm. For a 136 N applied hand force, the force in the IOL was 36 + 21 N. Advantages of this methodology include the determination of 3-D force vectors, especially those in the IOL, as well as computer generated 3-D visualization of results.

Low-profile versus conventional metacarpal plating systems: a comparison of construct stiffness and strength.

Smaller, lower-profile plates for metacarpal fixation may have the potential to reduce extensor tendon irritation and adhesions, but their sufficiency for stabilizing metacarpal fractures has not been studied. We investigated the relative stiffness and strength of low-profile and conventional plating systems. For apex dorsal bending (bending closed), no plates broke or had notable plastic deformation. The conventional plates exhibited higher overall bending rigidity than all other plates, but had a lower maximum bending moment than the smaller plates. In apex volar bending (bending open) and torsion, the conventional plates were remarkably more rigid and developed remarkably higher torque. In vivo metacarpal loads are primarily apex dorsal bending, and all plates performed well in this mode. Thus, the smaller, low-profile plates may be sufficient for metacarpal fixation, although patient compliance and the use of supplemental stabilization with a cast or splint should be considered.

Passive and active rehabilitation for partial lacerations of the canine flexor digitorum profundus tendon in zone II.

The purpose of this study was to compare the effect of unrestricted active versus passive mobilization on the gliding function and structural properties (ultimate load and stiffness) of repaired and nonrepaired canine flexor digitorum profundus tendons following partial laceration at 1 week. Using a radiographic method, normalized tendon gliding of the flexor digitorum profundus tendon adjacent to the metacarpal bone and total joint rotation were shown to be significantly greater in passive than in active tendons. Each group differed from their control group, however, by an average of only 5%. Both rehabilitation (active vs. passive) and treatment (repair vs. nonrepaired) of the partial tendon laceration significantly affected gap formation. Both active rehabilitation and repair of the laceration significantly increased gap formation compared with passive rehabilitation and nonrepair of the partial laceration. Rehabilitation did not significantly affect the normalized ultimate loads and stiffness in the passive and active groups but the nonrepair groups displayed significantly higher ultimate loads and stiffness than the repair groups.

Feasibility of partial A2 and A4 pulley excision: effect on finger flexor tendon biomechanics.

We investigated the effect of partial excision of the A2 and A4 digital pulleys, separately and in combination, on finger angular rotation and the energy for finger flexion. Statistically significant decreases in angular rotation resulted only after 50% and 75% excision of A2, A4, or A2 and A4 in combination. Work of flexion trends were weak and none of the changes were statistically significant. Although optimal finger function relies on the integrity of the A2 and A4 pulleys to maintain the efficiency of the digital flexor system, these data suggest that the A2 and A4 pulleys can be excised up to 25%, either separately or in combination, without significant effects on angular rotation. Decreases in total angular range of motion after 50% and 75% pulley excision were small, even for combined pulley excision (9 degrees +/- 3 degrees and 15 degrees +/- 5 degrees [mean +/- SD], respectively), and may be clinically acceptable.

Nerve wrapping with vein grafts in a rat model: a safe technique for the treatment of recurrent chronic compressive neuropathy.

The surgical outcome in recurrent chronic nerve compression remains unsatisfactory. Clinically, it has been reported that vein grafts can be used to wrap the nerve, following nerve decompression, to improve the functional recovery of the nerve. In order to determine the safety and feasibility of the technique of vein wrapping of nerve, this study assessed the effect of vein-graft wrapping around normal nerve. In each of 30 rats, a segment of femoral vein was wrapped around the sciatic nerve. Functional, electrophysiologic, and histologic testing was performed at 9, 12 and 15 weeks postoperatively, and the results showed no significant differences between the experimental and control groups. Neither scar tissue between the nerve and the vein graft, nor demyelinization or degeneration of the nerve fibers were identified histologically. The study demonstrated that autogenous vein-graft wrapping did not result in any pathologic nerve entrapment. The technique of vein wrapping of nerve appears to be safe, feasible, and reliable. The results supported further study for the use of vein-graft wrapping in the surgical management of recurrent chronic nerve compression.

Observations of convergence and uniqueness of node-based bone remodeling simulations.

Some investigators have indicated that mathematical theories and computational models of bone adaptation may not converge and that the density solutions from such simulations are dependent on the initial density distribution. In this study, two-dimensional finite element models were used to investigate the effect of initial density distribution on the final density distribution produced using a node-based bone remodeling simulation. The first model was a generic long bone, and the second was a proximal femur, For each model, we conducted time-dependent, node-based, linear rate-law bone remodeling simulations. Five initial density conditions were used with the generic long bone and three with the proximal femur. Remodeling simulations were performed, and the largest average nodal density differences at the end of the simulations were 0.000010 g/cm3 and 0.000006 g/cm3 for the generic long bone and proximal femur models, respectively. Results illustrate that, for a given set of loads and a given finite element model, the node-based bone adaptation algorithm can yield a unique density distribution. In conjunction with previous studies, this finding suggests that uniqueness of the density solution is dependent on both the mathematical theory and the computational implementation.

Different loads can produce similar bone density distributions.

Finite element models of a generic long bone and the proximal femur were used to identify important load characteristics and to determine whether small changes in load affect bone adaptation simulations. We also examined the effect of implants on the sensitivity of bone adaptation simulations to changes in loads. For each model, a primary load set was selected and incorporated in a bone adaptation simulation to generate a primary density distribution. A density-based load estimation method was used to determine a secondary set of loading conditions for each model. Each secondary load set was incorporated in a bone adaptation simulation and the resulting density distribution was compared to the corresponding primary density distribution. Nearly identical density distributions were produced for the natural generic long bone model (average nodal density difference 0.02 g/cm3). For the natural proximal femur model, the density distributions were very similar, but differences were apparent (average nodal density difference 0.07 g/cm3). The same primary and secondary load sets were used for bone adaptation simulations with implant models. For the proximal femur model, density distribution differences with the implant were very slightly less than those of the natural model. For the generic long bone model, the implant amplified differences between density distributions (average nodal density difference 0.14 g/cm3). Thus, variations in loading conditions may partially explain variations in long-term total joint outcome. The total equivalent stimulus load magnitudes for the two load sets for the generic long bone model were within 1%, and the stimulus-weighted average load directions were within 1 degree. The similarity of these parameters and the natural generic long bone density distributions indicate that the overall magnitude and average load direction are key factors affecting bone adaptation.

Computational method for determination of bone and joint loads using bone density distributions.

Because bone structure is influenced by mechanical loading during ontogeny, the geometry and density distribution of bones contain information about their loading histories. Based on a mathematical theory relating stress history to bone remodeling, we have developed a method to determine dominant bone loading conditions using an optimization procedure. We applied this load determination method using a simplified two-dimensional bone-end finite element model, for which a standard density distribution had been calculated under a given set of loading conditions. With this density distribution, the optimization procedure was used to determine the original loads from a broad set of many plausible basic load distributions and locations. The optimization procedure adjusted the magnitude of each basic load to achieve the desired tissue level attractor stress stimulus throughout the model. The results show that the density-based bone load determination method yields accurate results for basic test cases and, thus, may have potential for estimating in vivo bone loads for both extant and extinct animals.

Prodrugs of doxorubicin and melphalan and their activation by a monoclonal antibody-penicillin-G amidase conjugate.

The syntheses and cytotoxic activities of substituted N-phenylacetamido derivatives of doxorubicin and melphalan are described. The derivatives were designed as prodrugs which could be activated in a site-specific manner by monoclonal antibody-penicillin-G amidase (mAb-PGA) conjugates. N-(Phenylacetamido)doxorubicin (2) and N-(phenylacetyl)melphalan (6) were found to be 10- and 20-fold less cytotoxic against H2981 lung adenocarcinoma cells than doxorubicin and melphalan, respectively. When incubated with PGA, the cytotoxicity of 2 and 6 increased and became equivalent to that of the corresponding drugs from which they were made. The poor solubility characteristics of 2 in aqueous solutions provided the basis for the development of the more soluble doxorubicin derivatives, N-(4-aminophenylacetyl)doxorubicin (3) and N-(4-phosphonooxy)phenylacetyl)-doxorubicin (4). In vitro cytotoxicity assays indicated that 3 and 4 were at least 1000-fold less toxic than doxorubicin against H2981 cells. PGA and the mAb conjugate L6-PGA were able to effect the activation of 3 and 6 on H2981 cells (L6-antigen positive). Hydrolysis of the phosphate group of 4 was required prior to activation with PGA or L6-PGA. This was achieved using alkaline phosphatase, or by exposing 4 to phosphatases present in cell culture medium. The activation of 3, 4, and 6 on H2981 cells by L6-PGA occurred in an immunologically specific manner, since activation could be blocked by saturating cell surface antigens with L6 prior to treatment with L6-PGA. These results demonstrate that 3, 4, and 6 are prodrugs that can be specifically activated to release clinically approved anticancer agents by a mAb-PGA conjugate.

Equine tumor necrosis factor alpha: cloning and expression in Escherichia coli, generation of monoclonal antibodies, and development of a sensitive enzyme-linked immunosorbent assay.

We describe the production and purification of recombinant equine tumor necrosis factor alpha (rETNF alpha), generation and characterization of murine monoclonal antibodies (Mabs) and rabbit polyclonal antibodies (Pabs) against ETNF alpha, and development of a sensitive enzyme-linked immunosorbent assay (ELISA). Genomic-derived DNA sequences encoding mature ETNF alpha were reconstructed by the polymerase chain reaction (PCR) and oligonucleotide-directed mutagenesis and were cloned into the vector pFLAG-1 for expression in Escherichia coli. rETNF alpha was purified by anti-FLAG immunoaffinity chromatography and then used as immunogen for production of murine Mabs and rabbit Pabs. Three Mabs (6H4, 9B10, and 12F6) were obtained from one fusion. All three Mabs recognized rETNF alpha on western blots. Mabs 6H4 and 9B10 recognized similar epitopes on rENTF alpha and neutralized both rETNF alpha and native ETNF alpha (nETNF alpha) in a WEHI cell cytotoxicity assay. A sensitive ELISA was developed using Mab 6H4 and biotin-labeled rabbit Pabs. The ELISA was shown to detect levels of ENTF alpha as low as 100 pg/ml and was used to demonstrate the induction of ETNF alpha in horses with experimental endotoxemia. The rETNF alpha, antibodies, and ELISA developed in this report should be useful tools for studies of TNF-mediated diseases in horses.

N-(4'-hydroxyphenylacetyl)palytoxin: a palytoxin prodrug that can be activated by a monoclonal antibody-penicillin G amidase conjugate.

Palytoxin (PTX), one of the most toxic nonprotein molecules known, is cytotoxic at picomolar concentrations against a wide variety of cell types. In contrast to most cytotoxins, PTX exerts its activity extracellularly. A method for targeting PTX to tumor cells is described in which a monoclonal antibody-enzyme conjugate activates a PTX prodrug at surfaces of tumor cells. The prodrug, N-(4'-hydroxyphenylacetyl)palytoxin (NHPAP), was prepared by reacting PTX with an active ester of 4-hydroxyphenylacetic acid. NHPAP was 1000 times less toxic than PTX to a panel of carcinoma and lymphoma cell lines. The cytotoxic activity of the combination of penicillin G amidase from Escherichia coli with NHPAP was equal to PTX. Two cell lines that were multidrug resistant showed no enhanced resistance to NHPAP +/- penicillin G amidase. Immunologically specific activation of NHPAP took place when H2981 cells (L6 antigen positive) were treated with the monoclonal antibody conjugate L6-penicillin G amidase followed by NHPAP. This system is distinguished from other prodrug activation schemes, since the released drug exerts its activity extracellularly, has high potency, and may be able to overcome the multidrug resistant phenotype.

In vitro study of initial stability of a conical collared femoral component.

This in vitro experimental study compared the initial stability of an uncemented conical collared femoral component to that of the same component with the collar removed. The two configurations examined simulated joint resultant forces encountered in single leg stance and stair climbing. For the simulated single leg stance loads, the data do not allow any inferences about relative component stability. With the exception of one collarless control, all micromotion for single leg stance loading was under 150 microns, measured approximately 1.5 cm below the resection line. For scaled stair-climbing loads, however, the conical collared component group was significantly more stable than the collarless control group in transverse (primarily rotational) micromotion. The overall average measured transverse motion for the collarless control group was more than 3.7 times greater than that of the conical collar group at scaled stair-climbing loads. The two conical collared components loaded to full peak stair-climbing load (2,100 N) exhibited micromotion under 160 microns. The results suggest that the conical collar may improve stability of an uncemented prosthesis under loads that include an out-of-plane (rotational) component.

Phenylethanolamine N-methyltransferase in the brains of streptozotocin diabetic rats.

Recent evidence suggests that adrenergic neurons in the brainstem and hypothalamus are important for regulation of endocrine and cardiovascular function and the response to stress. Since abnormalities in several of these functions are observed in diabetic subjects, we investigated the effects of streptozotocin-induced diabetes on the activity of the enzyme that converts norepinephrine to epinephrine [phenylethanolamine N-methyltransferase (PNMT)] in the brainstem and hypothalamus of the rat. Enzyme activity was measured in the supernatant of sonicated brain tissues of saline-treated control, diabetic, and insulin-treated diabetic rats. One month after streptozotocin treatment, increases in plasma glucose and food intake occurred; these were restored to control levels by insulin treatment. PNMT activity was approximately 2-fold higher in the brainstem of diabetic rats than in controls (P less than 0.0001), and administration of insulin partially prevented the effects of diabetes on PNMT activity (P less than 0.01 compared to diabetics and P less than 0.05 compared to controls). Brainstem enzyme activity measured in all animals (n = 53) exhibited a significant correlation with plasma glucose concentrations (r = 0.51; P less than 0.001). Diabetes had no apparent effect on PNMT activity in the hypothalamus. These findings are the first to suggest that experimental diabetes alters PNMT activity in the medulla/pons.

Phospholipase A enzymes of Entamoeba histolytica: description and subcellular localization.

Cytolysis of target cells by Entamoeba histolytica is a rapid, contact-dependent event that appears to involve calcium and amebic phospholipase A enzymes. There are two phospholipase A enzymes of E. histolytica: one calcium independent and optimally active at an acid pH, and a second calcium dependent and most active at an alkaline pH. The amebic calcium-dependent enzyme was inhibited by pharmacological antagonists that have been shown to reduce cytolysis of target cells by intact amebae: phosphatidylcholine (10(-3) M), Rosenthal's inhibitor (dimethyl-dl-2,3-distearoylpropyl-2'-hydroxyethyl ammonium acetate; 10(-4) M), quinacrine (10(-4) M), and hydrocortisone (10(-4) M). Calcium-independent phospholipase A activity was inhibited by Rosenthal's inhibitor (10(-4) M) and quinacrine (10(-3) M). The calcium-dependent phospholipase A enzyme is highly associated with plasma membrane fractions; the calcium-independent enzyme is predominantly found in soluble fractions. These findings further suggest an association of calcium-dependent phospholipase A activity with cytolytic activity of E. histolytica.

[Fat embolism as a complication in shock. Clinical and therapeutic aspects (author's transl)]. / Fettembolie als Komplikation des Schockgeschehens. Klinische und therapeutische Aspekte.

In 36 patients the changes of the haemostasis in relation to hypovolaemic shock period were studied after polytrauma and or surgical operations. It was shown that a correlation between the hypovolaemic shock period and the degree of haemostatic disorder exists. The significance of this interaction for the manifestation of the fat embolism syndrome was demonstrated in two casuistics. It is concluded that the haemostatic disorder in shock enhances the development of the fat embolism syndrome. As a therapeutic consequence adequate volume substitution beside specific anticoagulation seems to be of utmost importance in the posttraumatic treatment.