Development and validation of a subject-specific moving-axis tibiofemoral joint model using MRI and EOS imaging during a quasi-static lunge

The aims of this study were to introduce and validate a novel computationally-efficient subject-specific tibiofemoral joint model. Subjects performed a quasi-static lunge while micro-dose radiation bi-planar X-rays (EOS Imaging, Paris, France) were captured at roughly 0°, 20°, 45°, 60°, and 90° of tibiofemoral flexion. Joint translations and rotations were extracted from this experimental data through 2D-to-3D bone reconstructions, using an iterative closest point optimization technique, and employed during model calibration and validation. Subject-specific moving-axis and hinge models for comparisons were constructed in the AnyBody Modeling System (AMS) from Magnetic Resonance Imaging (MRI)-extracted anatomical surfaces and compared against the experimental data. The tibiofemoral axis of the hinge model was defined between the epicondyles while the moving-axis model was defined based on two tibiofemoral flexion angles (0° and 90°) and the articulation modeled such that the tibiofemoral joint axis moved linearly between these two positions as a function of the tibiofemoral flexion. Outside this range, the joint axis was assumed to remain stationary. Overall, the secondary joint kinematics (ML: medial–lateral, AP: anterior-posterior, SI: superior-inferior, IE: internal-external, AA: adduction-abduction) were better approximated by the moving-axis model with mean differences and standard errors of (ML: −1.98 ± 0.37 mm, AP: 6.50 ± 0.82 mm, SI: 0.05 ± 0.20 mm, IE: 0.59 ± 0.36°, AA: 1.90 ± 0.79°) and higher coefficients of determination (R²) for each clinical measure. While the hinge model achieved mean differences and standard errors of (ML: −0.84 ± 0.45 mm, AP: 10.11 ± 0.88 mm, SI: 0.66 ± 0.62 mm, IE: −3.17 ± 0.86°, AA: 11.60 ± 1.51°).     

A novel method for measuring medial compartment pressures within the knee joint in-vivo

A novel method for the measurement of knee joint forces in-vivo is described. A thin (0.2mm) flexible electronic pressure sensor was inserted through a narrow arthroscopic portal into the osteoarthritic medial compartment of the knee joint. The sensor partially covered the load bearing area. The surgery was performed under local anaesthetic during normal arthroscopic examination following patient consent. Results are presented for 11 patients. The method was used in a pilot study to assess the effects of four valgus knee braces on medial compartment forces. An analysis of variance could not detect un-loading by any brace although there were large variations in force output. These variations may be attributable to shifts in the sensor position. In-vivo measurement of joint force is technically feasible.     

Quantifying in vivo laxity in the anterior cruciate ligament and individual knee joint structures

A custom knee loading apparatus (KLA), when used in conjunction with magnetic resonance imaging, enables in vivo measurement of the gross anterior laxity of the knee joint. A numerical model was applied to the KLA to understand the contribution of the individual joint structures and to estimate the stiffness of the anterior-cruciate ligament (ACL). The model was evaluated with a cadaveric study using an in situ knee loading apparatus and an ElectroForce test system. A constrained optimization solution technique was able to predict the restraining forces within the soft-tissue structures and joint contact. The numerical model presented here allowed in vivo prediction of the material stiffness parameters of the ACL in response to applied anterior loading. Promising results were obtained for in vivo load sharing within the structures. The numerical model overestimated the ACL forces by 27.61–92.71%. This study presents a novel approach to estimate ligament stiffness and provides the basis to develop a robust and accurate measure of in vivo knee joint laxity.     

Stem cell application for osteoarthritis in the knee joint: A minireview

Knee osteoarthritis is a chronic, indolent disease that will affect an ever increasing number of patients, especially the elderly and the obese. It is characterized by degeneration of the cartilage substance inside the knee which leads to pain, stiffness and tenderness. By some estimations in 2030, only in the United States, this medical condition will burden 67 million people. While conventional treatments like physiotherapy or drugs offer temporary relief of clinical symptoms, restoration of normal cartilage function has been difficult to achieve. Moreover, in severe cases of knee osteoarthritis total knee replacement may be required. Total knee replacements come together with high effort and costs and are not always successful. The aim of this review is to outline the latest advances in stem cell therapy for knee osteoarthritis as well as highlight some of the advantages of stem cell therapy over traditional approaches aimed at restoration of cartilage function in the knee. In addition to the latest advances in the field, challenges associated with stem cell therapy regarding knee cartilage regeneration and chondrogenesis in vitro and in vivo are also outlined and analyzed. Furthermore, based on their critical assessment of the present academic literature the authors of this review share their vision about the future of stem cell applications in the treatment of knee osteoarthritis.     

A novel functional calibration method for real-time elbow joint angles estimation with magnetic-inertial sensors

Magnetic-inertial measurement units (MIMUs) are often used to measure the joint angles between two body segments. To obtain anatomically meaningful joint angles, each MIMU must be computationally aligned (i.e., calibrated) with the anatomical rotation axes. In this paper, a novel four-step functional calibration method is presented for the elbow joint, which relies on a two-degrees-of-freedom elbow model. In each step, subjects are asked to perform a simple task involving either one-dimensional motions around some anatomical axes or a static posture. The proposed method was implemented on a fully portable wearable system, which, after calibration, was capable of estimating the elbow joint angles in real time. Fifteen subjects participated in a multi-session experiment that was designed to assess accuracy, repeatability and robustness of the proposed method. When compared against an optical motion capture system (OMCS), the proposed wearable system showed an accuracy of about 4° along each degree of freedom. The proposed calibration method was tested against different MIMU mountings, multiple repetitions and non-strict observance of the calibration protocol and proved to be robust against these factors. Compared to previous works, the proposed method does not require the wearer to maintain specific arm postures while performing the calibration motions, and therefore it is more robust and better suited for real-world applications.     

A method for quantifying the anterior load–displacement behavior of the human knee in both the low and high stiffness regions

The anterior load–displacement behavior of the human knee with an intact ACL is characterized by a very low stiffness region initially and a high stiffness region that develops as anterior load is increased. Although this behavior has been well recognized for some time, a method for quantitatively describing the behavior in these two regions based on limits of motion at specific values of anterior/posterior force has not yet been developed. Thus, the purposes of this study were to describe and justify such a method for measuring the laxity and stiffness in both of these regions in the intact knee.

Unique to this study, low stiffness and high stiffness laxities were computed based on three limits of motion for seven cadaveric knees tested at flexion angles ranging from 0° to 90°. Defining the reference position of the tibia relative to the femur, one limit was the 0 N posterior limit which was determined using a specially designed load cycle to reduce uncertainty in establishing a reference position. Defining the upper bound of the load–displacement curve, a second limit was the 225 N anterior limit. A third intermediate limit was the 45 N anterior limit, which was the load that represented the transition from the low stiffness to the high stiffness region. Stiffnesses corresponding to each of the two regions were computed using regression analysis and also estimated based on the laxities. Comparison between the computed and estimated stiffnesses demonstrated that the stiffnesses in both the low and high stiffness regions can be estimated reasonably accurately based on the laxities. Therefore, the 0 N posterior limit and the two laxities are the three quantities needed to describe the load–displacement behavior of the normal knee.     

Development and validation of a computational model of the knee joint for the evaluation of surgical treatments for osteoarthritis

A three-dimensional (3D) knee joint computational model was developed and validated to predict knee joint contact forces and pressures for different degrees of malalignment. A 3D computational knee model was created from high-resolution radiological images to emulate passive sagittal rotation (full-extension to 65°-flexion) and weight acceptance. A cadaveric knee mounted on a six-degree-of-freedom robot was subjected to matching boundary and loading conditions. A ligament-tuning process minimised kinematic differences between the robotically loaded cadaver specimen and the finite element (FE) model. The model was validated by measured intra-articular force and pressure measurements. Percent full scale error between FE-predicted and in vitro-measured values in the medial and lateral compartments were 6.67% and 5.94%, respectively, for normalised peak pressure values, and 7.56% and 4.48%, respectively, for normalised force values. The knee model can accurately predict normalised intra-articular pressure and forces for different loading conditions and could be further developed for subject-specific surgical planning.     

A novel and simple method of screening compounds for interaction with DNA: A validation study

We report the development of a simple, cost-effective assay for detecting compounds that have the ability to interact with and modify DNA. Potential uses for the assay lie in the areas of early genotoxicity testing of drug candidates, anticancer and antibiotic drug discovery, environmental monitoring and testing in the food, beverage and cosmetics industries. At present the assay has been used to assess direct-acting compounds only and it is yet to be established whether the assay is compatible with bio-activation. The methodology is based on the oxidative reaction of potassium permanganate with pyrimidine bases, which have become perturbed and more reactive by the agent under test. Results are recorded by use of UV/vis spectroscopy. The adaptation to a multi-well plate format provides the capacity for high throughput utilizing small amounts of compounds. Over 100 compounds, comprising different classes of DNA-binding chemicals as well as non-binding controls, have been put through the assay and the results compared with existing genotoxicity testing data from other methods. The assay has shown to be predictive of the results of other genotoxicity testing methods. We have found that the method is overall predictive of 71% of Ames bacterial reverse-mutation test results (where data are given) encompassing both negative and positive results.     

Anterior laxity,lateral tibial slope,and in situ ACL force differentiate knees exhibiting distinct patterns of motion during a pivoting event: A human cadaveric study

Knee instability following anterior cruciate ligament (ACL) rupture compromises function and increases risk of injury to the cartilage and menisci. To understand the biomechanical function of the ACL, previous studies have primarily reported the net change in tibial position in response to multiplanar torques, which generate knee instability. In contrast, we retrospectively analyzed a cohort of 13 consecutively tested cadaveric knees and found distinct motion patterns, defined as the motion of the tibia as it translates and rotates from its unloaded, initial position to its loaded, final position. Specifically, ACL-sectioned knees either subluxated anteriorly under valgus torque (VL-subluxating) (5 knees) or under a combination of valgus and internal rotational torques (VL/IR-subluxating) (8 knees), which were applied at 15 and 30° flexion using a robotic manipulator. The purpose of this study was to identify differences between these knees that could be driving the two distinct motion patterns. Therefore, we asked whether parameters of bony geometry and tibiofemoral laxity (known risk factors of non-contact ACL injury) as well as in situ ACL force, when it was intact, differentiate knees in these two groups. VL-subluxating knees exhibited greater sagittal slope of the lateral tibia by 3.6 ± 2.4° (p = 0.003); less change in anterior laxity after ACL-sectioning during a simulated Lachman test by 3.2 ± 3.2 mm (p = 0.006); and, at the peak applied valgus torque (no internal rotation torque), higher posteriorly directed, in situ ACL force by 13.4 ± 11.3 N and 12.0 ± 11.6 N at 15° and 30° of flexion, respectively (both p ≤ 0.03). These results may suggest that subgroups of knees depend more on their ACL to control lateral tibial subluxation in response to uniplanar valgus and multiplanar valgus and internal rotation torques as mediated by anterior laxity and bony morphology.     

Differing in vitro biology of equine,ovine, porcine and human articular chondrocytes derived from the knee joint: an immunomorphological study

For lack of sufficient human cartilage donors, chondrocytes isolated from various animal species are used for cartilage tissue engineering. The present study was undertaken to compare key features of cultured large animal and human articular chondrocytes of the knee joint. Primary chondrocytes were isolated from human, porcine, ovine and equine full thickness knee joint cartilage and investigated flow cytometrically for their proliferation rate. Synthesis of extracellular matrix proteins collagen type II, cartilage proteoglycans, collagen type I, fibronectin and cytoskeletal organization were studied in freshly isolated or passaged chondrocytes using immunohistochemistry and western blotting. Chondrocytes morphology, proliferation, extracellular matrix synthesis and cytoskeleton assembly differed substantially between these species. Proliferation was higher in animal derived compared with human chondrocytes. All chondrocytes expressed a cartilage-specific extracellular matrix. However, after monolayer expansion, cartilage proteoglycan expression was barely detectable in equine chondrocytes whereby fibronectin and collagen type I deposition increased compared with porcine and human chondrocytes. Animal-derived chondrocytes developed more F-actin fibers during culturing than human chondrocytes. With respect to proliferation and extracellular matrix synthesis, human chondrocytes shared more similarity with porcine than with ovine or equine chondrocytes. These interspecies differences in chondrocytes in vitro biology should be considered when using animal models.     

A novel method for measuring human hepatic lipase activity in postheparin plasma

The objective of this study was to establish a hepatic lipase (HL) assay method that can be applied to automatic clinical analyzers. Seventy-four hyperlipidemic subjects (men/women 45/29) were recruited. Lipase activity was assayed measuring the increase in absorbance at 546 nm due to quinonediimine dye production. Reaction mixture R-1 contained 50 mM Tris-HCl (pH 9.5), 0.5 mM glycerol-1,2-dioleate, 0.4% (unless otherwise noted) polyoxyethylene-nonylphenylether, 3 mM ATP, 3 mM MgCl(2), 1.5 mM CaCl(2), monoacylglycerol-specific lipase, glycerol kinase, glycerol-3-phosphate oxidase, 0.075% N,N-bis-(4-sulfobutyl)-3-methylaniline-2 Na, peroxidase, ascorbic acid oxidase. Reaction mixture R-2 contained 50 mM Tris-HCl (pH9.5), 0.15% 4-aminoantypirine. Automated assay for activity was performed with a Model 7080 Hitachi analyzer. In the lipase assay, 160 microl of R-1 was incubated at 37 degrees C with 3 microl of samples for 5 min, and 80 microl of R-2 was added. Within-run coefficient of variations was 0.9-1.0%. Calibration curve of lipase activity was linear (r = 0.999) between 0 and 320 U/l. Analytical recoveries of purified HL added to plasma were 96.6-99.8%. HL activity in postheparin plasma measured in this method had a closer correlation with HL mass by a sandwich ELISA (r = 0.888, P < 0.0001) than those in the conventional method using [(14)C-]triolein (r = 0.730, P < 0.0001). This assay method for HL activity can be applied to an automatic clinical analyzer.     

A novel sandwich immunosensing method for measuring cardiac troponin I in sera

Common methods for monitoring human cardiac troponin I (cTn I) are based on using antibodies against cTn I labeled with horseradish peroxidase, radioactive isotopes, or other labels. In this study, a novel label-free sandwich immunosensing method for measuring cTn I was developed. Three monoclonal antibodies (mAbs 9F5, 2F11, and 8C12) against human cTn I were generated by the commonly used hybridoma technique and characterized by a surface plasmon resonance (SPR) biosensor. An optimal pair of mAbs for measuring human cTn I was selected, as both mAbs have high affinities for cTn I and do not compete against each other for cTn I binding. An optical immunosensor for measuring cTn I in sera based on SPR was developed by using avidin as an intermediate layer and biotinylated-2F11 as the capturing antibody. Two detection methods for cTn I with the immunosensor were performed: (1) the direct detection of cTn I with a detection range of 2.5 to 40 microg/L and (2) the sandwich immunosensing method. In the sandwich assay mode, the second antibody 9F5 biologically amplified the sensor response. As a result, the sandwich assay showed a sensitivity of 0.25 microg/L and a detection range of 0.5 to 20 microg/L with within-run variation of 4.9 to 6.7% and between-run variation of 5.2 to 8.4%. This method has greatly enhanced the sensitivity for detection compared to that previously reported in the literatures.     

A novel validation and calibration method for motion capture systems based on micro-triangulation

Motion capture systems are widely used to measure human kinematics. Nevertheless, users must consider system errors when evaluating their results. Most validation techniques for these systems are based on relative distance and displacement measurements. In contrast, our study aimed to analyse the absolute volume accuracy of optical motion capture systems by means of engineering surveying reference measurement of the marker coordinates (uncertainty: 0.75 mm). The method is exemplified on an 18 camera OptiTrack Flex13 motion capture system. The absolute accuracy was defined by the root mean square error (RMSE) between the coordinates measured by the camera system and by engineering surveying (micro-triangulation). The original RMSE of 1.82 mm due to scaling error was managed to be reduced to 0.77 mm while the correlation of errors to their distance from the origin reduced from 0.855 to 0.209. A simply feasible but less accurate absolute accuracy compensation method using tape measure on large distances was also tested, which resulted in similar scaling compensation compared to the surveying method or direct wand size compensation by a high precision 3D scanner. The presented validation methods can be less precise in some respects as compared to previous techniques, but they address an error type, which has not been and cannot be studied with the previous validation methods.     

A novel in vitro method for the detection and characterization of photosensitizers

Photoactivation and binding of photoactive chemicals to proteins is a known prerequisite for the formation of immunogenic photoantigens and the induction of photoallergy. The intensive use of products and the availability of new chemicals, along with an increasing exposure to sun light contribute to the risk of photosensitizing adverse reactions. Dendritic cells (DC) play a pivotal role in the induction of allergic contact dermatitis. Human peripheral blood monocyte derived dendritic cells (PBMDC) were thus perceived as an obvious choice for the development of a novel in vitro photosensitization assay using the modulation of cell surface protein expression in response to photosensitizing agents. In this new protocol, known chemicals with photosensitizing, allergenic or non-allergenic potential were pre-incubated with PBMDCs prior to UVA irradiation (1 J/cm(2)). Following a 48 h incubation, the expression of the cell surface molecules CD86, HLA-DR and CD83 was measured by flow cytometry. All tested photosensitizers induced a significant and dose-dependent increase of CD86 expression after irradiation compared to non-irradiated controls. Moreover, the phototoxicity of the chemicals could also be determined. In contrast, (i) CD86 expression was not affected by the chosen irradiation conditions, (ii) increased CD86 expression induced by allergens was independent of irradiation and (iii) no PBMDC activation was observed with the non-allergenic control. The assay proposed here for the evaluation of the photoallergenic potential of chemicals includes the assessment of their allergenic, phototoxic and toxic potential in a single and robust test system and is filling a gap in the in vitro photoallergenicity test battery.     

东北主要树种光合作用可行的离体测定方法

树木光合作用的测定常因植株高大而难以开展, 其中离体测定是解决途径之一。但离体测定的方法及其可靠性因树种而异。选取东北东部温带森林中特性各异的7种主要树种: 针叶树(红松(Pinus koraiensis)、长白落叶松(Larix olgensis))、散孔材(白桦(Betula platyphylla)、胡桃楸(Juglans mandshurica))和环孔材(水曲柳(Fraxinus mandshurica)、黄榆(Ulmus macrocarpa)、蒙古栎(Quercus mongolica)), 首先采用光合速率恢复到光合诱导前稳定值90%的时间(T₉₀)长短和叶片蒸腾速率(E)的大小评估离体叶片水分供应状况及其光合活力, 以此确定较优的离体测定方案; 同时, 观测离体叶片的光合活力稳定时间; 最后通过比较原位测定和采用所确定的较优离体方案测定的各树种叶片气体交换参数, 论证采用离体测定光合作用的可靠性。结果表明: 除蒙古栎外的6个树种的离体叶片均具有较高、较稳定的水分供应和光合活力。离体枝条或复叶插入水中, 环剥去除切口处3 cm左右的韧皮部和剩余叶片的方法, 是这6个温带树种叶片光合能力的较优离体测定方法。6个树种叶片的T₉₀受树木特性的影响而差异显著(p < 0.05), 其中环孔材树种的T₉₀显著高于散孔材和针叶树种。6个树种离体叶片在1 h内均有较高、较稳定的水分供应和光合活力。在此期间离体所测得的绝大多数叶片的气体交换参数与其原位测定值之间的差异不显著。该研究提出了可行的树木叶片光合作用的离体测定方案, 适用于蒙古栎以外的其他6个温带树种。     

A novel non-invasive optical method for quantitative visualization of pH dynamics in the rhizosphere of plants

A novel optical method for non-invasive, quantitative and high-resolution imaging of spatial and temporal pH dynamics in soils mediated by plant roots is introduced. This method overcomes present limitations of measurement of pH, mainly short-term and punctiform measurements, by recording long-term dynamics of the micro-pattern of pH in the root-soil interface without disturbance of the biological and physico-chemical conditions. Juncus effusus L., rooting in a permanently flooded rhizotron, was selected as the test organism for qualifying the technique. The measurements showed pronounced diurnal variations of pH along the roots, particularly along the elongation zone. Diurnal oscillation of pH caused by the roots reached up to 0.5 units. Long-term records at 4 s intervals over more than 8 weeks revealed considerable spatial and temporal patterns of pH dynamics in the rhizosphere of about 10% of the pH scale (pH 7.0-8.5). The measured data were validated by the use of pH electrodes. Concomitantly measured oxygen concentration showed hypoxic conditions around root tips (10-70 micromol O2 L-1) and almost anoxic conditions (0.9 micromol O2 L-1) in the bulk soil. The present study qualifies this novel pH-sensing technique as a powerful analytical tool for quantitative visualization of undisturbed bioprocess dynamics.     

A novel method for measuring human lipoprotein lipase and hepatic lipase activities in postheparin plasma

The objective of this study was to establish a new lipoprotein lipase (LPL) and hepatic lipase (HL) activity assay method. Seventy normal volunteers were recruited. Lipase activities were assayed by measuring the increase in absorbance at 546 nm due to the quinoneine dye. Reaction mixture-1 (R-1) contained dioleoylglycerol solubilized with lauryldimethylaminobetaine, monoacylglycerol-specific lipase, glycerolkinase, glycerol-3-phosphate oxidase, peroxidase, ascorbic acid oxidase, and apolipoprotein C-II (apoC-II). R-2 contained Tris-HCl (pH 8.7) and 4-aminoantipyrine. Automated assay of lipase activities was performed with an automatic clinical analyzer. In the assay for HL + LPL activity, 160 microl R-1 was incubated at 37 degrees C with 2 microl of sample for 5 min, and 80 microl R-2 was added. HL activities were measured under the same conditions without apoC-II. HL and LPL activities were also measured by the conventional isotope method and for HL mass by ELISA. Lipase activity detected in a 1.6 M NaCl-eluted fraction from a heparin-Sepharose column was enhanced by adding purified apoC-II in a dose-dependent manner, whereas that eluted by 0.8 M NaCl was not. Postheparin plasma-LPL and HL activities measured in the present automated method had high correlations with those measured by conventional activity and mass methods. This automated assay method for LPL and HL activities is simple and reliable and can be applied to an automatic clinical analyzer.     

Accounting for post-randomization variables in meta-analysis: A joint meta-regression approach

Meta-regression is widely used in systematic reviews to investigate sources of heterogeneity and the association of study-level covariates with treatment effectiveness. Existing meta-regression approaches are successful in adjusting for baseline covariates, which include real study-level covariates (e.g., publication year) that are invariant within a study and aggregated baseline covariates (e.g., mean age) that differ for each participant but are measured before randomization within a study. However, these methods have several limitations in adjusting for post-randomization variables. Although post-randomization variables share a handful of similarities with baseline covariates, they differ in several aspects. First, baseline covariates can be aggregated at the study level presumably because they are assumed to be balanced by the randomization, while post-randomization variables are not balanced across arms within a study and are commonly aggregated at the arm level. Second, post-randomization variables may interact dynamically with the primary outcome. Third, unlike baseline covariates, post-randomization variables are themselves often important outcomes under investigation. In light of these differences, we propose a Bayesian joint meta-regression approach adjusting for post-randomization variables. The proposed method simultaneously estimates the treatment effect on the primary outcome and on the post-randomization variables. It takes into consideration both between- and within-study variability in post-randomization variables. Studies with missing data in either the primary outcome or the post-randomization variables are included in the joint model to improve estimation. Our method is evaluated by simulations and a real meta-analysis of major depression disorder treatments.     

A simple method for measuring colour in wild animals: validation and use on chest patch colour in geladas (Theropithecus gelada)

Adaptive hypotheses about colour variation are widespread in behavioural ecology, and several methods of objective colour assessment have been proposed and validated for use in a wide variety of taxa. However, to date, the most objective and reliable methods of assessing colour are not readily applied to wild animals. In the present study, we present a simple method for assessing colour in unrestrained, wild subjects using digital photography. The method we describe uses a digital camera, a colour standard, and colour analysis software, and can be used to measure any part of the visible colour spectrum. We demonstrate that the method: (1) is accurate and precise across different light conditions; (2) satisfies previous criteria regarding linearity and red, green, and blue equality; and (3) can be independently validated visually. In contrast with previous digital methods, this method can be used under natural light conditions and can be readily applied to subjects in their natural habitat. To illustrate this, we use the method to measure chest colour in wild geladas ( Theropithecus gelada ). Unique among primates, geladas have a red patch of skin on their chest and neck, which, for males, is thought to be a sexually selected signal. Offering some support to this hypothesis, we found differences in chest 'redness' for males across different age groups, with males in their reproductive prime exhibiting the reddest chests.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 231–240.