Development of a computational technique to measure cartilage contact area

Computational measurement of joint contact distributions offers the benefit of non-invasive measurements of joint contact without the use of interpositional sensors or casting materials. This paper describes a technique for indirectly measuring joint contact based on overlapping of articular cartilage computer models derived from CT images and positioned using in vitro motion capture data. The accuracy of this technique when using the physiological nonuniform cartilage thickness distribution, or simplified uniform cartilage thickness distributions, is quantified through comparison with direct measurements of contact area made using a casting technique. The efficacy of using indirect contact measurement techniques for measuring the changes in contact area resulting from hemiarthroplasty at the elbow is also quantified. Using the physiological nonuniform cartilage thickness distribution reliably measured contact area (ICC=0.727), but not better than the assumed bone specific uniform cartilage thicknesses (ICC=0.673). When a contact pattern agreement score (s_agree) was used to assess the accuracy of cartilage contact measurements made using physiological nonuniform or simplified uniform cartilage thickness distributions in terms of size, shape and location, their accuracies were not significantly different (p>0.05). The results of this study demonstrate that cartilage contact can be measured indirectly based on the overlapping of cartilage contact models. However, the results also suggest that in some situations, inter-bone distance measurement and an assumed cartilage thickness may suffice for predicting joint contact patterns.     

Electrical effects in bone

This paper presents a review of the work carried out on the electromechanical properties of bone over the the past three decades. Research in this field has established the piezoelectric nature of bone and identified collagen as the generating source in dry bone. Some of the characteristics of the strain generated potential (SGP) signal from dry and hydrated bone were found to be unaccountable in terms of a classical piezoelectric theory. Modifications of the theory were suggested and in the case of fully hydrated bone, a new mechanism (streaming potential) has emerged. The paper also reports on recent developments in the field and presents results from microstructural (osteonic) studies and from fluid-filled bone. The review indicates the need for actual in vivo work because most of the reported data were obtained, in the last decade, from in vitro work and were considered valid in vivo. Modelling of the mechanism which produces the SGP has been considered to explain the characteristics of these potentials. A representative model recently developed by the present authors and co-workers is reported. This model relates the generated potential to reorientation of spontaneous dipoles and differentiates between the generated and recorded signal, thus identifying effects from the measuring circuitry. The clinical aspects of electricity of bone in assisting fracture healing and the different techniques employed are mentioned briefly. Emphasis on new techniques of piezoelectric implants and their future development is also reported.     

Non-Invasive Drosophila ECG Recording by Using Eutectic Gallium-Indium Alloy Electrode: A Feasible Tool for Future Research on the Molecular Mechanisms Involved in Cardiac Arrhythmia

Background

Drosophila heart tube is a feasible model for cardiac physiological research. However, obtaining Drosophila electrocardiograms (ECGs) is difficult, due to the weak signals and limited contact area to apply electrodes. This paper presents a non-invasive Gallium-Indium (GaIn) based recording system for Drosophila ECG measurement, providing the heart rate and heartbeat features to be observed. This novel, high-signal-quality system prolongs the recording time of insect ECGs, and provides a feasible platform for research on the molecular mechanisms involved in cardiovascular diseases.

Methods

In this study, two types of electrode, tungsten needle probes and GaIn electrodes, were used respectively to noiselessly conduct invasive and noninvasive ECG recordings of Drosophila. To further analyze electrode properties, circuit models were established and simulated. By using electromagnetic shielded heart signal acquiring system, consisted of analog amplification and digital filtering, the ECG signals of three phenotypes that have different heart functions were recorded without dissection.

Results and Discussion

The ECG waveforms of different phenotypes of Drosophila recorded invasively and repeatedly with n value (n>5) performed obvious difference in heart rate. In long period ECG recordings, non-invasive method implemented by GaIn electrodes acts relatively stable in both amplitude and period. To analyze GaIn electrode, the correctness of GaIn electrode model established by this paper was validated, presenting accuracy, stability, and reliability.

Conclusions

Noninvasive ECG recording by GaIn electrodes was presented for recording Drosophila pupae ECG signals within a limited contact area and signal strength. Thus, the observation of ECG changes in normal and SERCA-depleted Drosophila over an extended period is feasible. This method prolongs insect survival time while conserving major ECG features, and provides a platform for electrophysiological signal research on the molecular mechanism involved in cardiac arrhythmia, as well as research related to drug screening and development.     

Rapid determination of L-lysine with an enzyme electrode by steady state and kinetic measurement

Fast and simple methods of determination of L-lysine by a potentiometric enzyme electrode based on a CO₂ electrode and L-lysine decarboxylase are described. Measuring devices for manual and automated operation for steady state response measurement as well as kinetic measurement are compared. Sample frequency may be increased by decreasing the time of a measuring cycle.     

Quantitative Imaging to Assess Tumor Response to Therapy: Common Themes of Measurement,Truth Data,and Error Sources

RATIONALE: Early detection of tumor response to therapy is a key goal. Finding measurement algorithms capable of early detection of tumor response could individualize therapy treatment as well as reduce the cost of bringing new drugs to market. On an individual basis, the urgency arises from the desire to prevent continued treatment of the patient with a high-cost and/or high-risk regimen with no demonstrated individual benefit and rapidly switch the patient to an alternative efficacious therapy for that patient. In the context of bringing new drugs to market, such algorithms could demonstrate efficacy in much smaller populations, which would allow phase 3 trials to achieve statistically significant decisions with fewer subjects in shorter trials. MATERIALS AND METHODS: This consensus-based article describes multiple, image modality-independent means to assess the relative performance of algorithms for measuring tumor change in response to therapy. In this setting, we describe specifically the example of measurement of tumor volume change from anatomic imaging as well as provide an overview of other promising generic analytic methods that can be used to assess change in heterogeneous tumors. To support assessment of the relative performance of algorithms for measuring small tumor change, data sources of truth are required. RESULTS: Very short interval clinical imaging examinations and phantom scans provide known truth for comparative evaluation of algorithms. CONCLUSIONS: For a given category of measurement methods, the algorithm that has the smallest measurement noise and least bias on average will perform best in early detection of true tumor change.     

Single-cell measurements of two-dimensional binding affinity across cell contacts

The two-dimensional (2D) affinity between protein molecules across contacting cells is a key parameter regulating and initiating several cellular processes. However, measuring 2D affinity can be challenging, and experimental data are limited. In addition, the obtained 2D affinities are typically averaged over the cell population. We here present a method to measure 2D affinity on single cells binding to polyhistidine-tagged fluorescent ligands anchored to a supported lipid bilayer (SLB). By decreasing the density of ligands in the SLB using imidazole, a new steady-state accumulation in the contact is obtained, and from this change, both the 2D affinity and the number of receptors on the cell can be determined. The method was validated on an SLB containing rat CD2 binding to the rat CD48 mutant T92A expressed on Jurkat T cells. The addition of imidazole did not influence the average 2D affinity (1/K_d), and the spread in affinities within the cell population was low, K_d = 4.9 ± 0.9 molecules/μm² (mean ± SD), despite an order of magnitude spread in ligand accumulation because of differences in receptor density. It was also found that cell contact size increased both with ligand density and with the number of receptors per cell but that the contact size stayed approximately constant when lowering the ligand density, above a density of around 10 rat CD2 molecules/μm², after the contact first had formed, indicative of a heterogeneous process. In summary, this method not only allows for single-cell affinities to be measured, but it can also reduce measurement and analysis time and improve measurement accuracy. Because of the low spread in 2D K_d within the cell population, the analysis can further be restricted to the cells showing the strongest binding, paving the way for using this method to study weak binding events.     

Optimal combination of electrodes and conductive gels for brain electrical impedance tomography

Background

Electrical impedance tomography (EIT) is an emerging imaging technology that has been used to monitor brain injury and detect acute stroke. The time and frequency properties of electrode–skin contact impedance are important for brain EIT because brain EIT measurement is performed over a long period when used to monitor brain injury, and is carried out across a wide range of frequencies when used to detect stroke. To our knowledge, no study has simultaneously investigated the time and frequency properties of both electrode and conductive gel for brain EIT.

Methods

In this study, the contact impedance of 16 combinations consisting of 4 kinds of clinical electrode and five types of commonly used conductive gel was measured on ten volunteers’ scalp for a period of 1 h at frequencies from 100 Hz to 1 MHz using the two-electrode method. And then the performance of each combination was systematically evaluated in terms of the magnitude of contact impedance, and changes in contact impedance with time and frequency.

Results

Results showed that combination of Ag⁺/Ag⁺Cl^? powder electrode and low viscosity conductive gel performed best overall (Ten 20^® in this study); it had a relatively low magnitude of contact impedance and superior performance regarding contact impedance with time (p?<?0.05) and frequency (p?<?0.05).

Conclusions

Experimental results indicates that the combination of Ag⁺/Ag⁺Cl^? powder electrode and low viscosity conductive gel may be the best choice for brain EIT.

     

Current-to-voltage convertor for measurement of oxygen

A highly sensitive, miniature, inexpensive circuit for the measurement of PO2 in vivo has been described. The circuit is constructed from a current-to-voltage convertor, clamping circuit, differential amplifier, and reverse voltage and overvoltage protector. The design of the circuit allows us to apply voltage bias to the measuring electrode while grounding the preparation. The clamping circuit holds the selected bias voltage constant while the differential amplifier subtracts this bias potential from the PO2 signal yielding an output voltage that is proportional to the current sensed by the oxygen electrode. The circuit is protected from reverse voltage and overvoltage.     

Three-Dimensional Quantitative Morphometric Analysis (QMA) for In Situ Joint and Tissue Assessment of Osteoarthritis in a Preclinical Rabbit Disease Model

This work utilises advances in multi-tissue imaging, and incorporates new metrics which define in situ joint changes and individual tissue changes in osteoarthritis (OA). The aims are to (1) demonstrate a protocol for processing intact animal joints for microCT to visualise relevant joint, bone and cartilage structures for understanding OA in a preclinical rabbit model, and (2) introduce a comprehensive three-dimensional (3D) quantitative morphometric analysis (QMA), including an assessment of reproducibility. Sixteen rabbit joints with and without transection of the anterior cruciate ligament were scanned with microCT and contrast agents, and processed for histology. Semi-quantitative evaluation was performed on matching two-dimensional (2D) histology and microCT images. Subsequently, 3D QMA was performed; including measures of cartilage, subchondral cortical and epiphyseal bone, and novel tibio-femoral joint metrics. Reproducibility of the QMA was tested on seven additional joints. A significant correlation was observed in cartilage thickness from matching histology-microCT pairs. The lateral compartment of operated joints had larger joint space width, thicker femoral cartilage and reduced bone volume, while osteophytes could be detected quantitatively. Measures between the in situ tibia and femur indicated an altered loading scenario. High measurement reproducibility was observed for all new parameters; with ICC ranging from 0.754 to 0.998. In conclusion, this study provides a novel 3D QMA to quantify macro and micro tissue measures in the joint of a rabbit OA model. New metrics were established consisting of: an angle to quantitatively measure osteophytes (σ), an angle to indicate erosion between the lateral and medial femoral condyles (ρ), a vector defining altered angulation (λ, α, β, γ) and a twist angle (τ) measuring instability and tissue degeneration between the femur and tibia, a length measure of joint space width (JSW), and a slope and intercept (m, Χ) of joint contact to demonstrate altered loading with disease progression, as well as traditional bone and cartilage and histo-morphometry measures. We demonstrate correlation of microCT and histology, sensitive discrimination of OA change and robust reproducibility.     

An international comparability study on quantification of total methyl cytosine content

A novel biocomposite film (MWCNTs-PNDGAChi), which contains multiwalled carbon nanotubes (MWCNTs) along with the incorporation of poly(nordihydroguaiaretic acid) and chitosan copolymer (PNDGAChi), has been synthesized on gold electrode by potentiostatic methods. The presence of MWCNTs in the biocomposite film enhances PNDGAChi’s surface coverage concentration (Γ) on the electrode and decreases degradation of PNDGAChi during cycling. The biocomposite film also exhibits promising enhanced electrocatalytic activity toward the oxidation of biochemical compounds such as epinephrine (EP) and norepinephrine (NEP). Cyclic voltammetry was used for the measurement of electroanalytical properties of analytes by means of MWCNTs-PNDGAChi biocomposite film modified gold electrode. The sensitivity values of MWCNTs-PNDGAChi biocomposite film modified gold electrode are higher than the values obtained for PNDGAChi film modified gold electrode. Electrochemical quartz crystal microbalance studies reveal the enhancements in the functional properties of MWCNTs and PNDGAChi present in MWCNTs-PNDGAChi biocomposite film. Surface morphology of the biocomposite films was studied using scanning electron microscopy, atomic force microscopy, and scanning tunneling microscopy. The surface morphology results reveal that PNDGAChi incorporated on MWCNTs. Finally, flow injection analysis was used for the amperometric detection of EP and NEP at MWCNTs-PNDGAChi film modified screen printed carbon electrode.     

Chloride-driven Electromechanical Phase Lags at Acoustic Frequencies Are Generated by SLC26a5, the Outer Hair Cell Motor Protein

Outer hair cells (OHC) possess voltage-dependent membrane bound molecular motors, identified as the solute carrier protein SLC26a5, that drive somatic motility at acoustic frequencies. The electromotility (eM) of OHCs provides for cochlear amplification, a process that enhances auditory sensitivity by up to three orders of magnitude. In this study, using whole cell voltage clamp and mechanical measurement techniques, we identify disparities between voltage sensing and eM that result from stretched exponential electromechanical behavior of SLC26a5, also known as prestin, for its fast responsiveness. This stretched exponential behavior, which we accurately recapitulate with a new kinetic model, the meno presto model of prestin, influences the protein’s responsiveness to chloride binding and provides for delays in eM relative to membrane voltage driving force. The model predicts that in the frequency domain, these delays would result in eM phase lags that we confirm by measuring OHC eM at acoustic frequencies. These lags may contribute to canceling viscous drag, a requirement for many models of cochlear amplification.     

Internal architecture of pampas deer antlers differs in males allocated with and without females

Bone mineralization of antlers and the depth of the antler seal (the basal surface of the cast antlers) are positively related to testosterone concentrations. Pampas deer males that are in permanent contact with females have greater, heavier, and darker antlers than males that are isolated from them. The objectives were to determine if antler compact/spongy bone ratio, antler seal depth, and compact bone darkness are greater in pampas deer males permanently allocated with females than those in males isolated from them. Antlers from males permanently allocated with or without females were cut transversally in seven points and scanned, and the compact/spongy ratio was calculated. The pixel intensity of each image was determined with a software for image analysis. The coronet of the antler was cut longitudinally, and the height of the most prominent protrusion was measured. The compact/spongy ratio was greater in antlers from males that were in contact with females in the second tine (P?=?0.02) and tended to be great in the two other tines (P?=?0.06 and P?=?0.1, respectively). Compact bone pixel color was darker in males in contact with females than that in males isolated from females in two points (P?=?0.02 and P?=?0.05, respectively) and tended to do so in two more (P?=?0.055 and P?=?0.1, respectively). Antler seal convexity was also greater in antlers from males in contact with females (P?=?0.006). We concluded that permanent contact with females stimulated pampas deer males increasing compact bone portion of the antler tines, the seal depth size, and the darkness of the compact bone.     

Double-mode impedance analysis of epithelial cell monolayers cultured on shear wave resonators

The viscoelastic behavior of epithelial cells (MDCK-I and MDCK-II) grown on AT-cut quartz crystals with a fundamental resonance at 5 MHz was investigated by impedance spectroscopy. Using the electromechanical model recently derived by Martin et al. [(1991) Anal Chem 63: 2272 – 2281] for Newtonian liquids in contact with shear wave resonators we quantified the viscous damping arising from the adherent cells by fitting the impedance data with a modified Butterworth-Van Dyke circuit in the region of the resonance frequency. Impedance spectroscopy was additionally performed in the frequency range from 1 Hz to 1 MHz to scrutinize the passive electrical properties of the epithelial cell layers using an additional platinum electrode. These data allow one to document the cell layers' integrity as well as the electrode coverage. We were able to confirm that the presence of a cell-layer mainly increases damping of the shear wave and does not exhibit a pure mass-load behavior. These findings were supported by the discovery that the inductance L in the electromechanical model was less influenced by the cell-layer than the resistance R. The apparent cell-viscosities determined by our method are 0.097 poise for MDCK-I and 0.142 poise for MDCK-II cell-layers. These low apparent viscosities may be explained in terms of a considerable spacing between the cells immobilized via their focal contacts and the quartz surface. Received: 5 June 1996 / Accepted: 6 August 1996     

A fluorometric method for the kinetic assay of indole-3-acetic acid oxidase.

The oxidation of IAA by peroxidase (1) and by more specific oxidases (2) leads to the formation of products which may have physiological activity (3, 4). The colorimetric estimation of IAA oxidation products involving reaction with p-dimethylaminocinnamaldehyde (DMACA) is reported to be more sensitive than other end point determinations such as the Salkowski and Ehrlich procedures which monitor the disappearance of IAA (5). These methods are end point procedures and, as such, are awkward and time consuming and present difficulties in obtaining kinetic data and measuring lag times. IAA oxidation has also been monitored by measuring ¹⁴CO₂ released from [1-¹⁴C] IAA (6) and uv spectral shifts during oxidation of IAA were reported by Meudt (3). The present paper reports a new procedure for the assay of horseradish peroxidase catalyzed oxidation of IAA. The assay procedure is based on the continuous measurement of a fluorescent product of the reaction.     

In-situ root extent measurements by electrical capacitance methods

A conceptual model is presented that provides a rational basis for using plant root capacitance as an in-situ measurement for assessing plant root development. This method is based on measuring the electricla capacitance of an equivalent parallel resistance-capacitance circuit formed by the interface between soil-water and the plant root surface. Nutrient solution studies using tomato (Lycopersicon esculentum Mill.) showed a good correlation between plant root capacitance and root mass. Stage of development studies showed plant root capacitance measurements capable of detecting root development rate and suggested the method to be sensitive to root function. Soil water content was shown to have a significant effect on plant root capacitance measurement. The possibility of using this technique to assess relative root function is discussed. Positioning of the plant shoot electrode was shown to also have a significant effect on measurement of plant root capacitance, demonstrating the need for using consistent measurement techniques. The electrical capacitance method shows considerable promise. More research is needed before it can be used routinely.     

Comparison of dynamic oxygen electrode methods for the measurement of KLa

The dynamic oxygen electrode method for measuring K_L a requires the use of a dynamic process model. Six models from the literature are described and compared with respect to their accuracy and ease of use. It is shown theoretically that for sufficient accuracy K_L a should be less than the inverse electrode response time. Experimental measurements demonstrate their application to viscous and nonviscous systems. The liquid diffusion film is shown to cause an important measurement lag that can be accounted for by a first-order time delay. Investigation on the influence of the experimental starting conditions show the importance of the gas and hold-up dynamics. A new method is proposed to simplify the K_L a calculation and to eliminate errors caused by starting conditions. This method, which accounts for gas, film, and electrode dynamical effects, requires only a simple semilog plot of the response data.     

Acute effect of whole-body vibration on electromechanical delay and vertical jump performance

Objectives:To determine if a change in vertical jump performance from acute whole-body vibration can be explained by indirectly assessing spindle sensitivity from electromechanical delay.Methods:Using a counter-balanced design, twenty college-aged participants performed whole-body vibration (WBV) and control treatments. WBV included 10 intervals (26 Hz, 3.6 mm) of 60 s in a half-squat followed by 60 s of rest. After 5 intervals, participants rested for 6-minutes before commencing the final 5 intervals. For the control, the exact same protocol of whole-body vibration was performed but without vibration. Electromechanical delay and vertical jump were assessed at baseline, during the 6-minute rest period and immediately after whole-body vibration and control.Results:There were no differences between treatments, for both electromechanical delay (F(2, 38)=1.385, p=0.263) and vertical jump (F(2, 38)=0.040, p<0.96). Whole-body vibration had no effect on vertical jump performance.Conclusion:The current whole-body vibration protocol is not effective for acute vertical jump or electromechanical delay enhancement. Also, since there was no effect on electromechanical delay, this suggests that whole-body vibration did not enhance muscle spindle sensitivity for the parameters examined.     

Development of a rapid culture method to induce adipocyte differentiation of human bone marrow-derived mesenchymal stem cells

Human mesenchymal stem cells (hMSCs) derived from bone marrow are multipotent stem cells that can regenerate mesenchymal tissues such as adipose, bone or muscle. It is thought that hMSCs can be utilized as a cell resource for tissue engineering and as human models to study cell differentiation mechanisms, such as adipogenesis, osteoblastogenesis and so on. Since it takes 2-3 weeks for hMSCs to differentiate into adipocytes using conventional culture methods, the development of methods to induce faster differentiation into adipocytes is required. In this study we optimized the culture conditions for adipocyte induction to achieve a shorter cultivation time for the induction of adipocyte differentiation in bone marrow-derived hMSCs. Briefly, we used a cocktail of dexamethasone, insulin, methylisobutylxanthine (DIM) plus a peroxisome proliferator-activated receptor γ agonist, rosiglitazone (DIMRo) as a new adipogenic differentiation medium. We successfully shortened the period of cultivation to 7-8 days from 2-3 weeks. We also found that rosiglitazone alone was unable to induce adipocyte differentiation from hMSCs in vitro. However, rosiglitazone appears to enhance hMSC adipogenesis in the presence of other hormones and/or compounds, such as DIM. Furthermore, the inhibitory activity of TGF-β1 on adipogenesis could be investigated using DIMRo-treated hMSCs. We conclude that our rapid new culture method is very useful in measuring the effect of molecules that affect adipogenesis in hMSCs.     

Measuring population-level plant gene flow with topological data analysis

Hybridization is an important factor influencing the evolution of many plant species. Measurement of interspecific gene flow and quantification of recombination provide insight into the processes that shape population genetic diversity. Recently, a new approach called topological data analysis (TDA) has begun to be implemented to track genomic recombination between species and populations. However, existing TDA methods tend to underestimate gene flow in cross-population models. Here, we present a novel graph-based approach to measuring gene flow, which occurs during ongoing speciation and hybridization in plants in various mating systems. We combine minimal spanning networks with topological filtering and then test the resulting parameter measurements on simulated primary divergence and secondary contact processes under different mating conditions. The resulting parameters, based on the first Betti number (total number of one-dimensional cycles), showed a high positive correlation with the number of migrants (N_m) in almost all cases of secondary contact. Considering the modularity of MSN networks, we have devised a parameter that works well in the event of primary divergence, where gene flow is at its highest level. This approach is suitable for various types of data, including SNPs, microsatellites, and binary restriction fragment data.