Expression and functional roles of angiopoietin-2 in skeletal muscles

Background

Angiopoietin-1 (ANGPT1) and angiopoietin-2 (ANGPT2) are angiogenesis factors that modulate endothelial cell differentiation, survival and stability. Recent studies have suggested that skeletal muscle precursor cells constitutively express ANGPT1 and adhere to recombinant ANGPT1 and ANGPT2 proteins. It remains unclear whether or not they also express ANGPT2, or if ANGPT2 regulates the myogenesis program of muscle precursors. In this study, ANGPT2 regulatory factors and the effects of ANGPT2 on proliferation, migration, differentiation and survival were identified in cultured primary skeletal myoblasts. The cellular networks involved in the actions of ANGPT2 on skeletal muscle cells were also analyzed.

Methodology/Principal Findings

Primary skeletal myoblasts were isolated from human and mouse muscles. Skeletal myoblast survival, proliferation, migration and differentiation were measured in-vitro in response to recombinant ANGPT2 protein and to enhanced ANGPT2 expression delivered with adenoviruses. Real-time PCR and ELISA measurements revealed the presence of constitutive ANGPT2 expression in these cells. This expression increased significantly during myoblast differentiation into myotubes. In human myoblasts, ANGPT2 expression was induced by H₂O₂, but not by TNFα, IL1β or IL6. ANGPT2 significantly enhanced myoblast differentiation and survival, but had no influence on proliferation or migration. ANGPT2-induced survival was mediated through activation of the ERK1/2 and PI-3 kinase/AKT pathways. Microarray analysis revealed that ANGPT2 upregulates genes involved in the regulation of cell survival, protein synthesis, glucose uptake and free fatty oxidation.

Conclusion/Significance

Skeletal muscle precursors constitutively express ANGPT2 and this expression is upregulated during differentiation into myotubes. Reactive oxygen species exert a strong stimulatory influence on muscle ANGPT2 expression while pro-inflammatory cytokines do not. ANGPT2 promotes skeletal myoblast survival and differentiation. These results suggest that muscle-derived ANGPT2 production may play a positive role in skeletal muscle fiber repair.     

A generic analytical foot rollover model for predicting translational ankle kinematics in gait simulation studies

The objective of this paper is to develop an analytical framework to representing the ankle–foot kinematics by modelling the foot as a rollover rocker, which cannot only be used as a generic tool for general gait simulation but also allows for case-specific modelling if required. Previously, the rollover models used in gait simulation have often been based on specific functions that have usually been of a simple form. In contrast, the analytical model described here is in a general form that the effective foot rollover shape can be represented by any polar function ρ=ρ(φ). Furthermore, a normalized generic foot rollover model has been established based on a normative foot rollover shape dataset of 12 normal healthy subjects. To evaluate model accuracy, the predicted ankle motions and the centre of pressure (CoP) were compared with measurement data for both subject-specific and general cases. The results demonstrated that the ankle joint motions in both vertical and horizontal directions (relative RMSE ～10%) and CoP (relative RMSE ～15% for most of the subjects) are accurately predicted over most of the stance phase (from 10% to 90% of stance). However, we found that the foot cannot be very accurately represented by a rollover model just after heel strike (HS) and just before toe off (TO), probably due to shear deformation of foot plantar tissues (ankle motion can occur without any foot rotation). The proposed foot rollover model can be used in both inverse and forward dynamics gait simulation studies and may also find applications in rehabilitation engineering.     

Dual functional roles of ATP in the human mitochondrial malic enzyme

Human mitochondrial malic enzyme is a regulatory enzyme with ATP as an inhibitor. Structural studies reveal that the enzyme has two ATP-binding sites, one at the NAD(+)-binding site in the active center and the other at the exo site in the tetramer interface. Inhibition of the enzyme activity is due to the competition between ATP and NAD(+) for the nucleotide-binding site at the active center with an inhibition constant of 81 microM. Binding of the ATP molecule at the exo site, on the other hand, is important for the maintenance of the quaternary structural integrity. The enzyme exists in solution at neutral pH and at equilibrium of the dimer and tetramer with a dissociation constant (K(TD)) of 0.67 microM. ATP, at a physiological concentration, shifts the equilibrium toward tetramer and decreases the K(TD) by many orders of magnitude. Mutation of a single residue Arg542 at the tetrameric interfacial exo site resulted in dimeric mutants. ATP thus has dual functional roles in the mitochondrial malic enzyme.     

Muscle mass in musculoskeletal models

Most current models of musculoskeletal dynamics lump a muscle's mass with its body segment, and then simulate the dynamics of these body segments connected by joints. As shown here, this popular approach leads to errors in the system's inertia matrix and hence in all aspects of the dynamics. Two simplified mathematical models were created to capture the relevant features of monoarticular and biarticular muscles, and the errors were analyzed. The models were also applied to two physiological examples: the triceps surae muscles that plantar flex the human ankle and the biceps femoris posterior muscle of the rat hind limb. The analysis of errors due to lumping showed that these errors can be large. Although the errors can be reduced in some postures, they cannot be easily eliminated in models that use segment lumping. Some options for addressing these errors are discussed.     

Quantitative functional anatomy of the lower limb with application to human gait

A scheme was developed to classify muscles according to their primary, secondary and tertiary functions, e.g. a muscle which produces primarily a flexion moment may also produce secondary abduction and tertiary internal rotation moments. The functions of muscles crossing the hip and knee joints were computed based upon the changing relative positions of joint centers and muscle origins and insertions during one gait cycle. The function of several of the major muscles crossing the hip and knee joints is reported for the different limb positions corresponding to normal gait. It was found that the amount of force necessary to produce a given moment about a joint was dependent upon the limb position. In addition, the muscle functions changed significantly with limb position. Electrical stimulation of muscles of a paralyzed subject gave qualitative support to the results.     

Two mathematical models for predicting dispersion of particles in the human lung

The dispersion of particles in the human lung is modeled as a series of virtual mixing tanks. Using the experimental results of Scherer et al. (1975, J. Appl. Physiol., 38(4), pp. 719-723) for a five-generation glass lung model, it is shown that each generation of the glass lung behaves like an independent virtual mixing tank. The corresponding resident time distribution is shown to have a variance approximately equal to the square of the average time a particle spends in the generation. By assuming that each generation of the human lung behaves as an independent virtual mixing tank, the realistic lung data provided by Weibel (1963, Morphometry of the Human Lung, Spinger-Verlag, New York) are used to validate this assumption in two ways. First, the half-width of the exhaled particle concentration profile is obtained. Second, a system of differential equations, with the concentration of particles in each mixing tank as its solution, is derived and solved numerically. This gives the exhaled concentration profile. Both techniques yield similar results to each other, and both give excellent agreement with the experimental data. The virtual mixing tank approach allows the complex mixing that occurs in the branching pathways of the lung to be more simply modeled. The model, thereby derived, is simple to change and could lead to enhancements in the understanding of the underlying processes contributing to the ventilation of the lung in health and disease.     

The functional classification of the human muscles that determine mandibular movements]

A new functional classification of muscles, determining movements of the mandible is presented. This classification reflects anatomo-topographic and histogenetic peculiarities of the muscles in question. Their functional differentiation in phylogenesis is motivated, coming from specific tasks they perform. Two groups of muscles of direct and mediated action on the mandible are distinguished. The groups are divided into functional subgroups. Mathematical correlations between various subgroups of the muscles at normal functioning of the mandible are presented. This makes possible to perform calculations of the acting muscle forces at planning reconstructive operative interventions in the maxillofacial area.     

Structures and functional roles of the sugar chains of human erythropoietins.

Erythropoietin (EPO) is a haemopoietic hormone specific to cells of erythroid lineage. EPO has recently become available for the treatment of anaemia as the first human recombinant biomedicine produced in heterologous mammalian cells. Human EPO is characterized by its large carbohydrate chains, which occupy close to 40% of its total mass. These sugar moieties were thought to be important for the biological activity of EPO, but detailed studies were not performed until the structures were elucidated. The variety of roles for the sugar chains were then immediately found once the structures were known. EPO is an excellent model for investigating the roles of sugar chains on glycoproteins, since its gene and its multiple glycoforms are available, as well as sensitive bioassays for testing. In this review, we will first summarize the known sugar chain structures of EPO from different host cells, and then discuss the host-cell dependent and peptide structure-dependent glycosylation of glycoproteins. We will then address how one investigates the roles of sugar chains of glycoproteins, show several examples of such investigations, and discuss the functional roles of HuEPO's sugar chains in its biosynthesis and secretion, its in vitro and in vivo biological activities, and its half-life in blood circulation.     

A technical method using musculoskeletal model to analyse dynamic properties of muscles during human movement

An effective way to avoid invading or injuring the subjects is to use the musculoskeletal model when studying the dynamic properties of muscles in vivo. So, we put forward a joint coordinate system-based method, which mainly focuses on the coordinate's transformation of corresponding muscle attachment points, respectively, in the model and the subject in order to reproduce the movement of the subject on the model. As muscle moment arm is usually used to evaluate the dynamic properties of muscles, the moment arms in elbow flexion for each of the major muscles crossing the elbow in 50 healthy subjects (25 males and 25 females), ranging in height from 1.50 to 1.80 m (mean 1.6542 m) are calculated and compared with the measured data obtained from anatomical studies reported in the literature. The trends of the value basically coincide with each other. So, this novel method can be valid.     

A technical method using musculoskeletal model to analyse dynamic properties of muscles during human movement

An effective way to avoid invading or injuring the subjects is to use the musculoskeletal model when studying the dynamic properties of muscles in vivo. So, we put forward a joint coordinate system-based method, which mainly focuses on the coordinate's transformation of corresponding muscle attachment points, respectively, in the model and the subject in order to reproduce the movement of the subject on the model. As muscle moment arm is usually used to evaluate the dynamic properties of muscles, the moment arms in elbow flexion for each of the major muscles crossing the elbow in 50 healthy subjects (25 males and 25 females), ranging in height from 1.50 to 1.80 m (mean 1.6542 m) are calculated and compared with the measured data obtained from anatomical studies reported in the literature. The trends of the value basically coincide with each other. So, this novel method can be valid.     

EFIN: predicting the functional impact of nonsynonymous single nucleotide polymorphisms in human genome

Background

Predicting the functional impact of amino acid substitutions (AAS) caused by nonsynonymous single nucleotide polymorphisms (nsSNPs) is becoming increasingly important as more and more novel variants are being discovered. Bioinformatics analysis is essential to predict potentially causal or contributing AAS to human diseases for further analysis, as for each genome, thousands of rare or private AAS exist and only a very small number of which are related to an underlying disease. Existing algorithms in this field still have high false prediction rate and novel development is needed to take full advantage of vast amount of genomic data.

Results

Here we report a novel algorithm that features two innovative changes: 1. making better use of sequence conservation information by grouping the homologous protein sequences into six blocks according to evolutionary distances to human and evaluating sequence conservation in each block independently, and 2. including as many such homologous sequences as possible in analyses. Random forests are used to evaluate sequence conservation in each block and to predict potential impact of an AAS on protein function. Testing of this algorithm on a comprehensive dataset showed significant improvement on prediction accuracy upon currently widely-used programs. The algorithm and a web-based application tool implementing it, EFIN (Evaluation of Functional Impact of Nonsynonymous SNPs) were made freely available (http://paed.hku.hk/efin/) to the public.

Conclusions

Grouping homologous sequences into different blocks according to the evolutionary distance of the species to human and evaluating sequence conservation in each group independently significantly improved prediction accuracy. This approach may help us better understand the roles of genetic variants in human disease and health.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-455) contains supplementary material, which is available to authorized users.     

Accuracy of gastrocnemius muscles forces in walking and running goats predicted by one-element and two-element Hill-type models

Hill-type models are commonly used to estimate muscle forces during human and animal movement—yet the accuracy of the forces estimated during walking, running, and other tasks remains largely unknown. Further, most Hill-type models assume a single contractile element, despite evidence that faster and slower motor units, which have different activation–deactivation dynamics, may be independently or collectively excited. This study evaluated a novel, two-element Hill-type model with “differential” activation of fast and slow contractile elements. Model performance was assessed using a comprehensive data set (including measures of EMG intensity, fascicle length, and tendon force) collected from the gastrocnemius muscles of goats during locomotor experiments. Muscle forces predicted by the new two-element model were compared to the forces estimated using traditional one-element models and to the forces measured in vivo using tendon buckle transducers. Overall, the two-element model resulted in the best predictions of in vivo gastrocnemius force. The coefficient of determination, r², was up to 26.9% higher and the root mean square error, RMSE, was up to 37.4% lower for the two-element model than for the one-element models tested. All models captured salient features of the measured muscle force during walking, trotting, and galloping (r²=0.26–0.51), and all exhibited some errors (RMSE=9.63–32.2% of the maximum in vivo force). These comparisons provide important insight into the accuracy of Hill-type models. The results also show that incorporation of fast and slow contractile elements within muscle models can improve estimates of time-varying, whole muscle force during locomotor tasks.     

The functional significance of morphological variation of the human mandible and masticatory muscles

A review is given of what is known about the functional significance of variation of the morphology of the human mandible and jaw muscles. First, the mandible is a lever transferring muscular forces to the teeth. The angle between corpus and ramus and the width of the ramus are particularly relevant in this respect as they determine the mechanical advantage of the lever system and the capacity for sagittal (open-close) movement. The stability of the mandible in asymmetric bites is especially affected by the ratio between the intermolar and intercondylar distances. The repertoire of bite forces that can be generated at any tooth and the loading pattern of the temporomandibular joint are strongly dependent on the relative size of the masseter, temporalis and medial pterygoid muscles. Second, executing its function as a lever, the mandible is subjected to shearing, bending and torsional forces. The bony parts harbouring the teeth, joints and muscle attachments serve to counter these forces; additional strength is needed in three areas i.e. in the symphysis, the condylar neck and in the transition area between corpus and ramus. In human populations there are clear-cut patterns of correlation between some facial skeletal traits, jaw joint morphology and strength and line of action of the jaw muscles. As a result, facial morphologies can be distinguished with marked differences in mechanical performance of their masticatory apparatus. It is suggested that they emerge as a result of diverging environmental influences during postnatal growth.     

Level of subject-specific detail in musculoskeletal models affects hip moment arm length calculation during gait in pediatric subjects with increased femoral anteversion

Biomechanical parameters of gait such as muscle's moment arm length (MAL) and muscle-tendon length are known to be sensitive to anatomical variability. Nevertheless, most studies rely on rescaled generic models (RGMo) constructed from averaged data of cadaveric measurements in a healthy adult population. As an alternative, deformable generic models (DGMo) have been proposed. These models integrate a higher level of subject-specific detail by applying characteristic deformations to the musculoskeletal geometry. In contrast, musculoskeletal models based on magnetic resonance (MR) images (MRMo) reflect the involved subject's characteristics in every level of the model. This study investigated the effect of the varying levels of subject-specific detail in these three model types on the calculated hip MAL during gait in a pediatric population of seven cerebral palsy subjects presenting aberrant femoral geometry. Our results show large percentage differences in calculated MAL between RGMo and MRMo. Furthermore, the use of DGMo did not uniformly reduce inter-model differences in calculated MAL. The magnitude of these percentage differences stresses the need to take these effects into account when selecting the level of subject-specific detail one wants to integrate in musculoskeletal. Furthermore, the variability of these differences between subjects and between muscles makes it very difficult to a priori estimate their importance for a biomechanical analysis of a certain muscle in a given subject.     

Muscle-specific indices to characterise the functional behaviour of human lower-limb muscles during locomotion

The mechanical output of a muscle may be characterised by having distinct functional behaviours, which can shift to satisfy the varying demands of movement, and may vary relative to a proximo-distal gradient in the muscle-tendon architecture (MTU) among lower-limb muscles in humans and other terrestrial vertebrates. We adapted a previous joint-level approach to develop a muscle-specific index-based approach to characterise the functional behaviours of human lower-limb muscles during movement tasks. Using muscle mechanical power and work outputs derived from experimental data and computational simulations of human walking and running, our index-based approach differentiated known distinct functional behaviours with varying mechanical demands, such as greater spring-like function during running compared with walking; with anatomical location, such as greater motor-like function in proximal compared with the distal lower-limb muscles; and with MTU architecture, such as greater strut-like muscles fibre function compared with the MTU in the ankle plantarflexors. The functional indices developed in this study provide distinct quantitative measures of muscle function in the human lower-limb muscles during dynamic movement tasks, which may be beneficial towards tuning the design and control strategies of physiologically-inspired robotic and assistive devices.     

Architectural and functional features of human triceps surae muscles during contraction

Architecturalproperties of the triceps surae muscles were determined in vivo for sixmen. The ankle was positioned at 15° dorsiflexion (15°)and 0, 15, and 30° plantar flexion, with the knee set at 0, 45, and90°. At each position, longitudinal ultrasonic images of the medial(MG) and lateral (LG) gastrocnemius and soleus (Sol) muscles wereobtained while the subject was relaxed (passive) and performed maximalisometric plantar flexion (active), from which fascicle lengths andangles with respect to the aponeuroses were determined. In the passivecondition, fascicle lengths changed from 59, 65, and 43 mm (knee,0°; ankle, 15°) to 32, 41, and 30 mm (knee, 90°ankle, 30°) for MG, LG, and Sol, respectively. Fascicle shorteningby contraction was more pronounced at longer fascicle lengths. MG hadgreatest fascicle angles, ranging from 22 to 67°, and was in a verydisadvantageous condition when the knee was flexed at 90°,irrespective of ankle positions. Different lengths and angles offascicles, and their changes by contraction, might be related todifferences in force-producing capabilities of the muscles and elasticcharacteristics of tendons and aponeuroses.

     

Structure of human estrone sulfatase suggests functional roles of membrane association

Estrone sulfatase (ES; 562 amino acids), one of the key enzymes responsible for maintaining high levels of estrogens in breast tumor cells, is associated with the membrane of the endoplasmic reticulum (ER). The structure of ES, purified from the microsomal fraction of human placentas, has been determined at 2.60-A resolution by x-ray crystallography. This structure shows a domain consisting of two antiparallel alpha-helices that protrude from the roughly spherical molecule, thereby giving the molecule a "mushroom-like" shape. These highly hydrophobic helices, each about 40 A long, are capable of traversing the membrane, thus presumably anchoring the functional domain on the membrane surface facing the ER lumen. The location of the transmembrane domain is such that the opening to the active site, buried deep in a cavity of the "gill" of the "mushroom," rests near the membrane surface, thereby suggesting a role of the lipid bilayer in catalysis. This simple architecture could be a prototype utilized by the ER membrane in dictating the form and the function of ER-resident enzymes.     

Spatiotemporal dynamics of two generic predator-prey models

We present the analysis of two reaction-diffusion systems modelling predator-prey interactions, where the predator displays the Holling type II functional response, and in the absence of predators, the prey growth is logistic. The local analysis is based on the application of qualitative theory for ordinary differential equations and dynamical systems, while the global well-posedness depends on invariant sets and differential inequalities. The key result is an L (∞)-stability estimate, which depends on a polynomial growth condition for the kinetics. The existence of an a priori L ( p )-estimate, uniform in time, for all p≥1, implies L (∞)-uniform bounds, given any nonnegative L (∞)-initial data. The applicability of the L (∞)-estimate to general reaction-diffusion systems is discussed, and how the continuous results can be mimicked in the discrete case, leading to stability estimates for a Galerkin finite-element method with piecewise linear continuous basis functions. In order to verify the biological wave phenomena of solutions, numerical results are presented in two-space dimensions, which have interesting ecological implications as they demonstrate that solutions can be 'trapped' in an invariant region of phase space.