A critical appreciation of pathway analysis in atherosclerotic disease. Cellular phenotypic plasticity as an illustrative example.

The rapid advancements in genome-scale (omics) techniques has created significant opportunities to investigate complex disease mechanisms in tissues and cells. Nevertheless, interpreting -omics data can be challenging, and pathway enrichment analysis is a frequently used method to identify candidate molecular pathways that drive gene expression changes. With a growing number of -omics studies dedicated to atherosclerosis, there has been a significant increase in studies and hypotheses relying on enrichment analysis. This brief review discusses the benefits and limitations of pathway enrichment analysis within atherosclerosis research. We highlight the challenges of identifying complex biological processes, such as cell phenotypic switching, within -omics data. Additionally, we emphasize the need for more comprehensive and curated gene sets that reflect the biological complexity of atherosclerosis. Pathway enrichment analysis is a valuable tool for gaining insights into the molecular mechanisms of atherosclerosis. Nevertheless, it is crucial to remain aware of the intrinsic limitations of this approach. By addressing these weaknesses, enrichment analysis in atherosclerosis can lead to breakthroughs in identifying the mechanisms of disease progresses, the identification of key driver genes, and consequently, advance personalized patient care.

Osteoarthritis year in review 2021: mechanics.

Osteoarthritis (OA) has a complex, heterogeneous and only partly understood etiology. There is a definite role of joint cartilage pathomechanics in originating and progressing of the disease. Although it is still not identified precisely enough to design or select targeted treatments, the progress of this year's research demonstrates that this goal became much closer. On multiple scales - tissue, joint and whole body - an increasing number of studies were done, with impressive results. (1) Technology based instrument innovations, especially when combined with machine learning models, have broadened the applicability of biomechanics. (2) Combinations with imaging make biomechanics much more precise & personalized. (3) The combination of Musculoskeletal & Finite Element Models yield valid personalized cartilage loads. (4) Mechanical outcomes are becoming increasingly meaningful to inform and evaluate treatments, including predictive power from biomechanical models. Since most recent advancements in the field of biomechanics in OA are at the level of a proof op principle, future research should not only continue on this successful path of innovation, but also aim to develop clinical workflows that would facilitate including precision biomechanics in large scale studies. Eventually this will yield clinical tools for decision making and a rationale for new therapies in OA.

Botulinum toxin injections minimally affect modelled muscle forces during gait in children with cerebral palsy.

BACKGROUND: Children with cerebral palsy (CP) present altered gait patterns and electromyography (EMG) activity compared to typically developing children. To temporarily reduce muscular activity and to correct the abnormal muscle force balance, Botulinum Toxin type A (BTX-A) injections are used. RESEARCH QUESTION: What is the effect of BTX-A injections on dynamic muscle forces during gait, when calculated using an EMG-constrained approach?. METHODS: Retrospective data of ten typically developing (TD) and fourteen children with spastic diplegic CP were used for musculoskeletal modeling and dynamic simulations of gait, before and after BTX-A treatment. Individual muscle forces were calculated using an EMG-constrained optimization, in which EMG of eight muscles was used as muscle excitation signal to constrain the muscle activation patterns. Paired t-tests were used to compare average modelled muscle forces in different phases of the gait cycle pre- and post-BTX-A, summarized in the muscle profile score. Two-sample t-tests were used to determine significant differences between TD and pre- and post-BTX-A modelled muscle forces. RESULTS: For most muscles, the force was decreased in CP compared to TD children in all phases of the gait cycle, both before and after BTX-A treatment. Differences in muscle forces before and after BTX-A treatment were limited, with only few significant differences between pre- and post-BTX-A. Compared to a standard static optimization approach, imposing the EMG activity increased modelled muscle forces for most muscles. SIGNIFICANCE: Our findings indicate that BTX-A treatment has a limited effect on the muscle balance in CP children. Besides that, the use of EMG-constrained optimization is recommended when studying muscle balance in children with CP.

Discriminant validity of 3D joint kinematics and centre of mass displacement measured by inertial sensor technology during the unipodal stance task.

BACKGROUND: The unipodal stance task is a clinical task that quantifies postural stability and alignment of the lower limb joints, while weight bearing on one leg. As persons with knee osteoarthritis (KOA) have poor postural and knee joint stability, objective assessment of this task might be useful. OBJECTIVE: To investigate the discriminant validity of three-dimensional joint kinematics and centre of mass displacement (COM) between healthy controls and persons with knee KOA, during unipodal stance using inertial sensors. Additionally, the reliability, agreement and construct validity are assessed to determine the reproducibility and accuracy of the discriminating parameters. METHODS: Twenty healthy controls and 19 persons with unilateral severe KOA were included. Five repetitions of the unipodal stance task were simultaneously recorded by an inertial sensor system and a camera-based system (gold standard). Statistical significant differences in kinematic waveforms between healthy controls and persons with severe knee KOA were determined using one-dimensional statistical parametric mapping (SPM1D). RESULTS: Persons with severe knee KOA had more lateral trunk lean towards the contralateral leg, more hip flexion throughout the performance of the unipodal stance task, more pelvic obliquity and COM displacement towards the contralateral side. However, for the latter two parameters the minimum detectable change was greater than the difference between healthy controls and persons with severe knee KOA. The construct validity was good (coefficient of multiple correlation 0.75, 0.83 respectively) and the root mean squared error (RMSE) was low (RMSE <1.5°) for the discriminant parameters. CONCLUSION: Inertial sensor based movement analysis can discriminate between healthy controls and persons with severe knee KOA for lateral trunk lean and hip flexion, but unfortunately not for the knee angles. Further research is required to improve the reproducibility and accuracy of the inertial sensor measurements before they can be used to assess differences in tasks with a small range of motion.

Single-event multilevel surgery, but not botulinum toxin injections normalize joint loading in cerebral palsy patients.

BACKGROUND: Many patients with cerebral palsy present a pathologic gait pattern, which presumably induces aberrant musculoskeletal loading that interferes with natural bone growth, causing bone deformations on the long term. Botulinum toxin interventions and single-event multilevel surgeries are used to restore the gait pattern, assuming that a normal gait pattern restores musculoskeletal loading and thus prevents further bone deformation. However, it is unknown if these interventions are able to restore musculoskeletal loading. Hence, we investigated the impact of botulinum toxin injections and single-event multilevel surgery on musculoskeletal loading. METHODS: Gait data collected in 93 children with bilateral cerebral palsy, which included pre- and post multi-level botulinum toxin (49 children) and single-event multilevel surgery (44 children) assessments, and 15 typically developing children were retrospectively processed using a musculoskeletal modelling workflow to calculate joint angles, moments, muscle and joint contact force magnitudes and orientations. Differences from the typically developing waveform were expressed by a root-mean square difference were compared using paired t-tests for each intervention separately (alpha <0.05). FINDINGS: Botulinum toxin induced significant changes in the joint angles, but did not improve the muscle and joint contact forces. Single-event multilevel surgery induced significant kinematic and kinetic changes, which were associated with improved muscle and joint contact forces. INTERPRETATION: The present results indicate that botulinum toxin injections were not able to restore normal gait kinematics nor musculoskeletal loading, whereas single-event multilevel surgery did successfully restore both. Therefore, single-event multilevel surgery might be protective against the re-occurrence of bone deformation on the longer term.

A musculoskeletal model of the hand and wrist: model definition and evaluation.

To improve our understanding on the neuromechanics of finger movements, a comprehensive musculoskeletal model is needed. The aim of this study was to build a musculoskeletal model of the hand and wrist, based on one consistent data set of the relevant anatomical parameters. We built and tested a model including the hand and wrist segments, as well as the muscles of the forearm and hand in OpenSim. In total, the model comprises 19 segments (with the carpal bones modeled as one segment) with 23 degrees of freedom and 43 muscles. All required anatomical input data, including bone masses and inertias, joint axis positions and orientations as well as muscle morphological parameters (i.e. PCSA, mass, optimal fiber length and tendon length) were obtained from one cadaver of which the data set was recently published. Model validity was investigated by first comparing computed muscle moment arms at the index finger metacarpophalangeal (MCP) joint and wrist joint to published reference values. Secondly, the muscle forces during pinching were computed using static optimization and compared to previously measured intraoperative reference values. Computed and measured moment arms of muscles at both index MCP and wrist showed high correlation coefficients (r = 0.88 averaged across all muscles) and modest root mean square deviation (RMSD = 23% averaged across all muscles). Computed extrinsic flexor forces of the index finger during index pinch task were within one standard deviation of previously measured in-vivo tendon forces. These results provide an indication of model validity for use in estimating muscle forces during static tasks.

The morphological and molecular mechanisms of epithelial/endothelial-to-mesenchymal transition and its involvement in atherosclerosis.

Cell transdifferentiation occurs during cardiovascular development or remodeling either as a pathologic feature in the progression of disease or as a response to injury. Endothelial-to-Mesenchymal Transition (EndMT) is a process that is classified as a specialized form of Epithelial-to-Mesenchymal Transition (EMT), in which epithelial cells lose their epithelial characteristics and gain a mesenchymal phenotype. During transdifferentiation, cells lose both cell-cell contacts and their attachment to the basement membrane. Subsequently, the shape of the cells changes from a cuboidal to an elongated shape. A rearrangement of actin filaments facilitates the cells to become motile and prime their migration into the underlying tissue. EMT is a key process during embryonic development, wound healing and tissue regeneration, but has also been implicated in pathophysiological processes, such organ fibrosis and tumor metastases. EndMT has been associated with additional pathophysiological processes in cardiovascular related diseases, including atherosclerosis. Recent studies prove a significant role for EndMT in the progression and destabilization of atherosclerotic plaques, as a consequence of EndMT-derived fibroblast infiltration and the increased secretion of matrix metalloproteinase respectively. In this review we will discuss the essential molecular and morphological mechanisms of EMT and EndMT, along with their common denominators and key differences. Finally, we will discuss the role of EMT/EndMT in developmental and pathophysiological processes, focusing on the potential role of EndMT in atherosclerosis in more depth.

Influence of plasticization time, curing conditions, storage time, and core properties on the drug release from Aquacoat-coated pellets.

Theophylline or chlorpheniramine maleate pellets were coated with an aqueous ethylcellulose dispersion, Aquacoat. The influence of the plasticization time, curing conditions, storage time, and core properties on the drug release were investigated. The plasticization time (time between plasticizer addition to the polymer dispersion and the spraying process) did not affect the drug release, when the water-soluble plasticizer triethyl citrate, was used because of its rapid uptake by the colloidal polymer particles. In contrast, with the water-insoluble plasticizer acetyltributyl citrate (ATBC), plasticization time (1/2 h vs 24 h) influenced the drug release, the longer plasticization time resulted in a slower drug release because of a more complete plasticizer uptake prior to the coating step. However a thermal aftertreatment of the coated pellets at eleylated temperatures (curing step) reduced/eliminated the effect of the plasticization time with ATBC. In general, curing reduced the drug release and resulted in stable drug release profiles. The time period between the coating and the curing step was not critical when the pellets were cured for a longer time. The structure of the pellet core (high dose matrix vs low dose layered pellet) strongly affected the drug release. A slow, zero-order drug release was obtained with high dose theophylline pellets, while a more rapid, first-order release pattern was obtained with low dose theophylline-layered nonpareil pellets.

Drug release from beads coated with an aqueous colloidal ethylcellulose dispersion, Aquacoat, or an organic ethylcellulose solution.

The objective was to investigate several factors (composition of the coating formulation, the type and pH of the release medium and curing conditions), which influence the drug release from beads coated with either the aqueous ethylcellulose dispersion, Aquacoat or an organic ethylcellulose solution. The chlorpheniramine maleate release from Aquacoat-coated beads was faster in pH 7.4 buffer than in 0.1 N HCl. Increasing the curing time and curing temperature decreased the drug release in pH 7.4 buffer but did not affect the release in 0.1 N HCl. In contrast, the drug release from beads coated with the ethanolic ethylcellulose solution was not affected by the curing step, the release medium or the addition of sodium lauryl sulfate. Scanning electron microscopy and contact angle measurements explained the release data. The differences in the drug release behavior of aqueous--and organic solvent--ethylcellulose--coated beads could be attributed to the differences in the film formation process.

Tackiness of acrylic and cellulosic polymer films used in the coating of solid dosage forms.

The objective was to determine the tackiness of acrylic and cellulosic polymer films in order to make predictions on the tackiness (agglomeration) of coated dosage forms during coating and curing. Force-displacement curves of the detachment process of two polymeric films were used as a measure of tackiness. Various polymers (cellulosic (Aquacoat and acrylics (Eudragit RS 30D, L 30D, NE 30D)), plasticizers (triacetin, triethyl citrate, tributyl citrate, acetyltributyl citrate) and anti-tacking agents (talc and glyceryl monostearate) were investigated. The order of tackiness for films prepared from the different aqueous polymer dispersions was in order of Eudragit NE 30D > RS 30D > RL 30D > Aquacoat. The tackiness increased with increasing plasticizer concentration due to the softening of the polymer. A correlation between the minimum film formation temperature and the tackiness was observed, however, no correlation between the tackiness and the lipophilicity of the plasticizer was seen. Talc and glyceryl monostearate (GMS) reduced the tackiness of the films significantly, with GMS being effective at much lower concentrations. Curing of Eudragit RS 30D-coated theophylline beads at temperatures higher than 40 degrees C in an irreversible agglomeration of the beads and damage of the coating upon separation of the beads. This resulted in a faster release than with uncured beads. Blending the beads with talc just prior to the curing step eliminated the agglomeration and therefore film damage, even at a curing temperature of 60 degrees C.

Multiprocessor and memory architecture of the neurocomputer SYNAPSE-1.

A general purpose neurocomputer, SYNAPSE-1, which exhibits a multiprocessor and memory architecture is presented. It offers wide flexibility with respect to neural algorithms and a speed-up factor of several orders of magnitude--including learning. The computational power is provided by a 2-dimensional systolic array of neural signal processors. Since the weights are stored outside these NSPs, memory size and processing power can be adapted individually to the application needs. A neural algorithms programming language, embedded in C(+2) has been defined for the user to cope with the neurocomputer. In a benchmark test, the prototype of SYNAPSE-1 was 8000 times as fast as a standard workstation.