Modelling motor units in 3D: influence on muscle contraction and joint force via a proof of concept simulation

Functional heterogeneity is a skeletal muscle’s ability to generate diverse force vectors through localised motor unit (MU) recruitment. Existing 3D macroscopic continuum-mechanical finite element (FE) muscle models neglect MU anatomy and recruit muscle volume simultaneously, making them unsuitable for studying functional heterogeneity. Here, we develop a method to incorporate MU anatomy and information in 3D models. Virtual fibres in the muscle are grouped into MUs via a novel “virtual innervation” technique, which can control the units’ size, shape, position, and overlap. The discrete MU anatomy is then mapped to the FE mesh via statistical averaging, resulting in a volumetric MU distribution. Mesh dependency is investigated using a 2D idealised model and revealed that the amount of MU overlap is inversely proportional to mesh dependency. Simultaneous recruitment of a MU’s volume implies that action potentials (AP) propagate instantaneously. A 3D idealised model is used to verify this assumption, revealing that neglecting AP propagation results in a slightly less-steady force, advanced in time by approximately 20 ms, at the tendons. Lastly, the method is applied to a 3D, anatomically realistic model of the masticatory system to demonstrate the functional heterogeneity of masseter muscles in producing bite force. We found that the MU anatomy significantly affected bite force direction compared to bite force magnitude. MU position was much more efficacious in bringing about bite force changes than MU overlap. These results highlight the relevance of MU anatomy to muscle function and joint force, particularly for muscles with complex neuromuscular architecture.

     

Autotaxin delays apoptosis induced by carboplatin in ovarian cancer cells

Drug resistance remains a barrier to the effective long term treatment of ovarian cancer. We have established an RNAi-based screen to identify genes which confer resistance to carboplatin or paclitaxel. To validate the screen we showed that siRNA interfering with the apoptosis regulators FLIP and Bcl-X_L conferred sensitivity to paclitaxel and carboplatin respectively. The expression of 90 genes which have previously been shown to be over-expressed in drug-resistant ovarian cancer was inhibited using siRNA and the impact on sensitivity to carboplatin and paclitaxel was assessed. ENPP2 was identified as a candidate gene causing drug resistance. ENPP2 encodes autotaxin, a phospholipase involved in the synthesis of the survival factor lysophosphatidic acid. siRNA directed to ENPP2 resulted in earlier apoptosis following treatment with carboplatin. 2-carbacyclic phosphatidic acid (ccPA 16:1), a small molecule inhibitor of autotaxin, also accelerated apoptosis induced by carboplatin. Stable ectopic expression of autotaxin in OVCAR-3 cells led to a delay in apoptosis. When serum was withdrawn to remove exogenous LPA, ccPA caused a pronounced potentiation of apoptosis induced by carboplatin in cells expressing autotaxin. These results indicate that autotaxin delays apoptosis induced by carboplatin in ovarian cancer cells.     

A life cycle assessment of packaging options for contrast media delivery: comparing polymer bottle vs. glass bottle

Purpose

This paper compares environmental impacts of two packaging options for contrast media offered by GE Healthcare: +PLUSPAK? polymer bottle and traditional glass bottle. The study includes all relevant life cycle stages from manufacturing to use and final disposal of the bottles and includes evaluation of a variety of end-of-life disposal scenarios. The study was performed in accordance with the international standards ISO 14040/14044, and a third-party critical review was conducted.

Methods

The functional unit is defined as the packaging of contrast media required to deliver one dose of 96 mL to a patient for an X-ray procedure. Primary data are from GE Healthcare and its suppliers; secondary data are from the ecoinvent database and the literature. A variety of end-of-life disposal scenarios are explored using both cutoff and market-based allocation. Impact assessment includes human health (midpoint) and ecosystems and resources (end point) categories from ReCiPe (H) and cumulative energy demand. Sensitivity analyses include (1) bottle size, (2) secondary packaging, (3) manufacturing electricity, (4) glass recycled content, (5) scrap rate, (6) distribution transport, (7) contrast media, and (8) choice of impact assessment method. Uncertainty analysis is performed to determine how data quality affects the study conclusions.

Results and discussion

This study indicates that the polymer bottle outperforms the glass bottle in every environmental impact category considered. Bottle components are the most significant contributors, and the vial body has the highest impacts among bottle components for both polymer and glass bottles. The polymer bottle exhibits lower impact in all impact categories considered regardless of the following: end-of-life treatment (using either cutoff or market-based allocation), bottle size, manufacturing electricity grid mix, glass recycled content, scrap rate, contrast media, distribution transport (air vs. ocean), and choice of impact assessment method. Secondary packaging can be a major contributor to impact. The polymer bottle has considerably lower impact compared to the glass bottle for all multi-pack configurations, but the comparison is less clear for single-pack configurations due to significantly higher packaging material used per functional dose, resulting in proportionally higher impacts in all impact categories.

Conclusions

The lower impacts of the polymer bottle for this packaging application can be attributed to lower material and manufacturing impacts, lower distribution impacts, and lower end-of-life disposal impacts. The results of this study suggest that using polymer rather than glass bottles provides a means by which to lower environmental impact of contrast media packaging.     

The BH3 Mimetic Obatoclax Accumulates in Lysosomes and Causes Their Alkalinization

Obatoclax belongs to a class of compounds known as BH3 mimetics which function as antagonists of Bcl-2 family apoptosis regulators. It has undergone extensive preclinical and clinical evaluation as a cancer therapeutic. Despite this, it is clear that obatoclax has additional pharmacological effects that contribute to its cytotoxic activity. It has been claimed that obatoclax, either alone or in combination with other molecularly targeted therapeutics, induces an autophagic form of cell death. In addition, obatoclax has been shown to inhibit lysosomal function, but the mechanism of this has not been elucidated. We have evaluated the mechanism of action of obatoclax in eight ovarian cancer cell lines. Consistent with its function as a BH3 mimetic, obatoclax induced apoptosis in three cell lines. However, in the remaining cell lines another form of cell death was evident because caspase activation and PARP cleavage were not observed. Obatoclax also failed to show synergy with carboplatin and paclitaxel, chemotherapeutic agents which we have previously shown to be synergistic with authentic Bcl-2 family antagonists. Obatoclax induced a profound accumulation of LC-3 but knockdown of Atg-5 or beclin had only minor effects on the activity of obatoclax in cell growth assays suggesting that the inhibition of lysosomal function rather than stimulation of autophagy may play a more prominent role in these cells. To evaluate how obatoclax inhibits lysosomal function, confocal microscopy studies were conducted which demonstrated that obatoclax, which contains two basic pyrrole groups, accumulates in lysosomes. Studies using pH sensitive dyes demonstrated that obatoclax induced lysosomal alkalinization. Furthermore, obatoclax was synergistic in cell growth/survival assays with bafilomycin and chloroquine, two other drugs which cause lysosomal alkalinization. These studies explain, for the first time, how obatoclax inhibits lysosomal function and suggest that lysosomal alkalinization contributes to the cytotoxic activity of obatoclax.     

A novel computational method to determine subject-specific bite force and occlusal loading during mastication

The evaluation of three-dimensional occlusal loading during biting and chewing may assist in development of new dental materials, in designing effective and long-lasting restorations such as crowns and bridges, and for evaluating functional performance of prosthodontic components such as dental and/or maxillofacial implants. At present, little is known about the dynamic force and pressure distributions at the occlusal surface during mastication, as these quantities cannot be measured directly. The aim of this study was to evaluate subject-specific occlusal loading forces during mastication using accurate jaw motion measurements. Motion data was obtained from experiments in which an individual performed maximal effort dynamic chewing cycles on a rubber sample with known mechanical properties. A finite element model simulation of one recorded chewing cycle was then performed to evaluate the deformation of the rubber. This was achieved by imposing the measured jaw motions on a three-dimensional geometric surface model of the subject’s dental impressions. Based on the rubber’s deformation and its material behaviour, the simulation was used to compute the resulting stresses within the rubber as well as the contact pressures and forces on the occlusal surfaces. An advantage of this novel modelling approach is that dynamic occlusal pressure maps and biting forces may be predicted with high accuracy and resolution at each time step throughout the chewing cycle. Depending on the motion capture technique and the speed of simulation, the methodology may be automated in such a way that it can be performed chair-side. The present study demonstrates a novel modelling methodology for evaluating dynamic occlusal loading during biting or chewing.