Theoretical response of a bifurcating axon with a locally altered axial resistivity

A short region of high axial resistivity at one daughter branch of an axonal bifurcation can produce frequency dependent differential conduction of action potentials. The increase in resistivity need be only a few times that in the rest of the axon and length of the affected region need be only a fraction of a resting length constant (based on the local value of axial resistivity). The typical pattern observed will be that the unaffected daughter branch will conduct action potentials from the parent axon normally at all frequencies of stimulation, but the branch with the high resistance region will only follow action potentials within a restricted frequency range. In that band-pass region, the branch may conduct nearly all or only a small percentage of the action potentials from the parent axon.     

Isolated periodic solutions of the Hodgkin-Huxley equations

Periodic solutions of the current clamped Hodgkin-Huxley equations (Hodgkin & Huxley, 1952 J. Physiol. 117, 500) that arise by degenerate Hopf bifurcation were studied recently by Labouriau (1985 SIAM J. Math. Anal. 16, 1121, 1987 Degenerate Hopf Bifurcation and Nerve Impulse (Part II), in press). Two parameters, temperature T and sodium conductance gNa were varied from the original values obtained by Hodgkin & Huxley. Labouriau's work proved the existence of small amplitude periodic solution branches that do not connect locally to the stationary solution branch, and had not been previously computed. In this paper we compute these solution branches globally. We find families of isolas of periodic solutions (i.e. branches not connected to the stationary branch). For values of gNa in the range measured by Hodgkin & Huxley, and for physically reasonable temperatures, there are isolas containing orbitally asymptotically stable solutions. The presence of isolas of periodic solutions suggests that in certain current space clamped membrane experiments, action potentials could be observed even though the stationary state is stable for all current stimuli. Once produced, such action potentials will disappear suddenly if the current stimulus is either increased or decreased past certain values. Under some conditions, "jumping" between action potentials of different amplitudes might be observed.     

Distribution of Cell Adhesion Molecules (CAM) along the Branching Neurite Surface: Model-Inherited Effects of the Branch Diameters and Mode of CAM Transport

In many cases, an increase in the surface density of cell adhesion molecules (CAM) in the distal parts of a growing neurite is favorable for the neurite elongation. This increase is attained by exocytotic insertion of CAM-containing vesicles into the growth cones with subsequent redistribution of CAM along the cell surface due to lateral diffusion and endocytosis. Using a mathematical model describing these processes, we quantitatively describe conditions providing two qualitatively different profiles in a branching neurite: (i) the CAM surface density increases along both daughter branches, which would be in favor of further outgrowth of both branches, i.e., successful branching, or (ii) the CAM surface density increases along one daughter branch and decreases along another branch, which could lead to the retraction of the latter. The geometric factors and mechanisms underlying the intracellular CAM transport to the daughter growth cones were proved to determine the profile of CAM surface density. A similarity in the diameters of daughter branches, their short lengths, a high value of the lateral transfer constant, and partitioning of CAM transport at the branching point proportionally to the surface areas of daughter branches are in favor of an increase in the CAM surface density along both daughter branches. Asymmetric branching can lead to a decrease in the CAM surface density along the thinner or thicker daughter branch, if CAM trafficking was equally partitioned or was proportional to the branch cross-sectional areas, respectively. The proposed model helps to understand possible relationships between the intracellular CAM trafficking, CAM surface distribution, and geometry of branching of the neurites.     

Ca channel gating during cardiac action potentials.

How do Ca channels conduct Ca ions during the cardiac action potential? We attempt to answer this question by applying a two-microelectrode technique, previously used for Na and K currents, in which we record the patch current and the action potential at the same time (Mazzanti, M., and L. J. DeFelice. 1987. Biophys. J. 12:95-100, and 1988. Biophys. J. 54:1139-1148; Wellis, D., L. J. DeFelice, and M. Mazzanti. 1990. Biophys. J. 57:41-48). In this paper, we also compare the action currents obtained by the technique with the step-protocol currents obtained during standard voltage-clamp experiments. Individual Ca channels were measured in 10 mM Ca/1 Ba and 10 mM Ba. To describe part of our results, we use the nomenclature introduced by Hess, P., J. B. Lansman, and R. W. Tsien (1984. Nature (Lond.). 311:538-544). With Ba as the charge carrier, Ca channel kinetics convert rapidly from long to short open times as the patch voltage changes from 20 to -20 mV. This voltage-dependent conversion occurs during action potentials and in step-protocol experiments. With Ca as the charge carrier, the currents are brief at all voltages, and it is difficult to define either the number of channels in the patch or the conductance of the individual channels. Occasionally, however, Ca-conducting channels spontaneously convert to long-open-time kinetics (in Hess et al., 1984, notation, mode 2).(ABSTRACT TRUNCATED AT 250 WORDS)     

Degenerate Hopf bifurcation and isolated periodic solutions of the Hodgkin-Huxley model with varying sodium ion concentration

Points of degenerate Hopf bifurcation in the Hodgkin-Huxley model are found as parameters temperature T and voltage level of sodium VNa are varied. Local techniques of degenerate Hopf bifurcation analysis are used to show the existence of families of periodic solutions of the model: isolated branches of periodic solutions (i.e. branches not connected to the stationary branch) are found in addition to Hopf branches. Purely numerical techniques are used to show that the isolas persist for VNa up to a value slightly greater than 114 mV. Under some conditions there are multiple stable periodic solutions, so "jumping" between action potentials of different amplitudes might be observed.     

蕨类植物rbcL基因正选择和负选择位点的鉴定

基于分支模型、位点模型及分支-位点模型对蕨类的rbcL基因所受到的选择压力进行了分析.结果显示:分支模型下检测到大部分分支处于负选择,仅4个分支处于正选择压力下,并且仅2分支具有统计上的显著性;在位点模型下,通过比较模型M1a/M2a和M7/M8,在氨基酸水平上模型M2a和模型M8均鉴定出98L位点被正选择;在模型M8下,鉴定负向选择位点共228个,占总序列的83.82%,从而揭示出负选择对rbcL基因的进化起着非常重要的作用;在分支位点-模型c下鉴定出262A被正选择.98L、262A位点分别位于rbcL羧基末端α/β桶结构域的第3和第8个α螺旋上.蕨类通过该结构域的适应性进化,适应白垩纪被子植物兴起而引发的陆地生态系统改变,研究结果为以后实验分析提供了首选位点.     

Inhibition in branched afferent neurons of the bullfrog tongue

Summary Single fibers of the bullfrog glossopharyngeal nerve give rise to several peripheral branches, each innervating separate fungiform papillae on the dorsal surface of the tongue. Extracellular electrodes were used to stimulate and record simultaneously from several papillae and from the central branch.Minor changes in centrally recorded neural output were caused by collision of action potentials originating in separate branches of a common fiber.Following an antidromic or orthodromic action potential in any branch, a series of excitability changes occured in that branch. Normal excitability was regained within 5 msec of an action potential, but was followed by a secondary decrease in excitability, which reached a minimum approximately 50 msec after the spike, and returned to normal within 200–400 msec after the spike. Subthreshold stimuli caused no depression, while doubling the stimulus strength above threshold did not enhance depression. After several spikes, both amplitude and duration of depression increased. Depression could be evoked even after the gustatory receptors were surgically removed.Post-stimulus depression in fiber branches is suggested as one source of gustatory adaptation, and may also contribute to interference between stimulating substances.The authors are particularly grateful for assistance and advice from Dr. Douglas Junge, of the School of Dentistry and Department of Physiology at the University of California, Los Angeles. The reported work was supported by NIDR Contract No. 69-2227 to Dr. Junge, and was carried out while one of the authors (JAM) held a PHS postdoctoral traineeship with the Department of Zoology, U.C.L.A., and the other (MSB) held a NIH predoctoral traineeship with the Department of Anatomy, U.C.L.A. Draughts of the paper have been read and criticized by Dr. Junge and Dr. J. P. Leader, of Auckland University.     

Outward sodium current in beating heart cells.

This article is a study of the fast Na current during action potentials. We have investigated the outward Na current (Mazzanti, M., and L.J. DeFelice. 1987. Biophys. J. 52:95-100) in more detail, and we have asked whether it goes through the same channels associated with the rapid depolarization phase of action potentials. We address the question by patch clamping single, spontaneously beating, embryonic chick ventricle cells, using two electrodes to record the action potential and the patch current simultaneously. The chief limitation is the capacitive current, and in this article we describe a new method to subtract it. Varying the potential and the Na concentration in the patch pipette, and fitting the corrected currents to a standard model (Ebihara, L., and E.A. Johnson. 1980. Biophys. J. 32:779-790), provides evidence that the outward current is carried by the same channels that conduct the inward current. We compare the currents in beating cells to currents in nonbeating cells using whole-cell and cell-attached patch clamp recordings. The latter tend to show more positive Na reversal potentials, with the implication that internal Na is higher in beating cells. We propose that the plateau of the action potential, which is partly due to an inward Ca current, exceeds Na action current reversal potentials, and that this driving force gives rise to an outward movement of Na ions. The existence of such a current would imply that the fast repolarization phase after the upstroke of cardiac action potentials is partly due to the Na action current.     

Tree Branching: Leonardo da Vinci's Rule versus Biomechanical Models

This study examined Leonardo da Vinci''s rule (i.e., the sum of the cross-sectional area of all tree branches above a branching point at any height is equal to the cross-sectional area of the trunk or the branch immediately below the branching point) using simulations based on two biomechanical models: the uniform stress and elastic similarity models. Model calculations of the daughter/mother ratio (i.e., the ratio of the total cross-sectional area of the daughter branches to the cross-sectional area of the mother branch at the branching point) showed that both biomechanical models agreed with da Vinci''s rule when the branching angles of daughter branches and the weights of lateral daughter branches were small; however, the models deviated from da Vinci''s rule as the weights and/or the branching angles of lateral daughter branches increased. The calculated values of the two models were largely similar but differed in some ways. Field measurements of Fagus crenata and Abies homolepis also fit this trend, wherein models deviated from da Vinci''s rule with increasing relative weights of lateral daughter branches. However, this deviation was small for a branching pattern in nature, where empirical measurements were taken under realistic measurement conditions; thus, da Vinci''s rule did not critically contradict the biomechanical models in the case of real branching patterns, though the model calculations described the contradiction between da Vinci''s rule and the biomechanical models. The field data for Fagus crenata fit the uniform stress model best, indicating that stress uniformity is the key constraint of branch morphology in Fagus crenata rather than elastic similarity or da Vinci''s rule. On the other hand, mechanical constraints are not necessarily significant in the morphology of Abies homolepis branches, depending on the number of daughter branches. Rather, these branches were often in agreement with da Vinci''s rule.     

The plasmamembrane calmodulin-dependent calcium pump: a major regulator of nitric oxide synthase I

The plasma membrane calcium/calmodulin-dependent calcium ATPase (PMCA) (Shull, G.E., and J. Greeb. 1988. J. Biol. Chem. 263:8646-8657; Verma, A.K., A.G. Filoteo, D.R. Stanford, E.D. Wieben, J.T. Penniston, E.E. Strehler, R. Fischer, R. Heim, G. Vogel, S. Mathews, et al. 1988. J. Biol. Chem. 263:14152-14159; Carafoli, E. 1997. Basic Res. Cardiol. 92:59-61) has been proposed to be a regulator of calcium homeostasis and signal transduction networks of the cell. However, little is known about its precise mechanisms of action. Knock-out of (mainly neuronal) isoform 2 of the enzyme resulted in hearing loss and balance deficits due to severe inner ear defects, affecting formation and maintenance of otoconia (Kozel, P.J., R.A. Friedman, L.C. Erway, E.N. Yamoah, L.H. Liu, T. Riddle, J.J. Duffy, T. Doetschman, M.L. Miller, E.L. Cardell, and G.E. Shull. 1998. J. Biol. Chem. 273:18693-18696). Here we demonstrate that PMCA 4b is a negative regulator of nitric oxide synthase I (NOS-I, nNOS) in HEK293 embryonic kidney and neuro-2a neuroblastoma cell models. Binding of PMCA 4b to NOS-I was mediated by interaction of the COOH-terminal amino acids of PMCA 4b and the PDZ domain of NOS-I (PDZ: PSD 95/Dlg/ZO-1 protein domain). Increasing expression of wild-type PMCA 4b (but not PMCA mutants unable to bind PDZ domains or devoid of Ca2+-transporting activity) dramatically downregulated NO synthesis from wild-type NOS-I. A NOS-I mutant lacking the PDZ domain was not regulated by PMCA, demonstrating the specific nature of the PMCA-NOS-I interaction. Elucidation of PMCA as an interaction partner and major regulator of NOS-I provides evidence for a new dimension of integration between calcium and NO signaling pathways.     

Propagation of action potentials in squid giant axons. Repetitive firing at regions of membrane inhomogeneities

Effects of reduction in potassium conductance on impulse conduction were studied in squid giant axons. Internal perfusion of axons with tetraethylammonium (TEA) ions reduces G K and causes the duration of action potential to be increased up to 300 ms. This prolongation of action potentials does not change their conduction velocity. The shape of these propagating action potentials is similar to membrane action potentials in TEA. Axons with regions of differing membrane potassium conductances are obtained by perfusing the axon trunk and one of its two main branches with TEA after the second branch has been filled with normal perfusing solution. Although the latter is initially free of TEA, this ion diffuses in slowly. Up until a large amount of TEA has diffused into the second branch, action potentials in the two branches have very different durations. During this period, membrane regions with prolonged action potentials are a source of depolarizing current for the other, and repetitive activity may be initiated at transitional regions. After a single stimulus in either axon region, interactions between action potentials of different durations usually led to rebound, or a short burst, of action potentials. Complex interactions between two axon regions whose action potentials have different durations resembles electric activity recorded during some cardiac arrhythmias.     

Simultaneous imaging of cell and mitochondrial membrane potentials.

The distribution of charged membrane-permeable molecular probes between intracellular organelles, the cytoplasm, and the outside medium is governed by the relative membrane electrical potentials of these regions through coupled equilibria described by the Nernst equation. A series of highly fluorescent cationic dyes of low membrane binding and toxicity (Ehrenberg, B., V. Montana, M.-D. Wei, J. P. Wuskell, and L. M. Loew, 1988. Biophys. J. 53:785-794) allows the monitoring of these equilibria through digital imaging video microscopy. We employ this combination of technologies to assess, simultaneously, the membrane potentials of cells and of their organelles in situ. We describe the methodology and optimal conditions for such measurements, and apply the technique to concomitantly follow, with good time resolution, the mitochondrial and plasma membrane potentials in several cultured cell lines. The time course of variations induced by chemical agents (ionophores, uncouplers, electron transport, and energy transfer inhibitors) in either or both these potentials is easily quantitated, and in accordance with mechanistic expectations. The methodology should therefore be applicable to the study of more subtle and specific, biologically induced potential changes in cells.     

Purified N-cadherin is a potent substrate for the rapid induction of neurite outgrowth

N-cadherin is the predominant mediator of calcium-dependent adhesion in the nervous system (Takeichi, M. 1988. Development (Camb.). 102: 639-655). Investigations using antibodies to block N-cadherin function (Bixby, J.L., R.L. Pratt, J. Lilien, and L.F. Reichardt. 1987. Proc. Natl. Acad. Sci. USA. 84:2555-2569; Bixby, J.L., J. Lilien, and L.F. Reichardt. 1988. J. Cell Biol. 107:353-362; Tomaselli, K.J., K.N. Neugebauer, J.L. Bixby, J. Lilien, and L.F. Reichardt. 1988. Neuron. 1:33-43) or transfection of the N-cadherin gene into heterologous cell lines (Matsunaga, M., K. Hatta, A. Nagafuchi, and M. Takeichi. 1988. Nature (Lond.). 334:62-64) have provided evidence that N-cadherin, alone or in combination with other molecules, can participate in the induction of neurite extension. We have developed an affinity purification procedure for the isolation of whole N-cadherin from chick brain and have used the isolated protein as a substrate for neurite outgrowth. N-cadherin promotes the rapid extension of neurites from chick ciliary ganglion neurons, which extend few or no neurites on adhesive but noninducing substrates such as polylysine, tissue culture plastic, and collagens. N-cadherin is extremely potent, more so than the L1 adhesion molecule, and comparable to the extracellular matrix protein laminin. Compared to laminin, however. N-cadherin promotes outgrowth from ciliary ganglion neurons extremely rapidly and with a distinct morphology. These results provide a direct demonstration that N-cadherin is sufficient to induce neurite outgrowth when substrate bound and suggest that the mechanism(s) involved may differ from that induced by laminin.     

Redistribution of water via layering branches between connected parent and daughter trees in Norway spruce clonal groups

Key message

We measured sap flow and shoot water potentials in clonally connected parent and daughter trees. We found bidirectional flow patterns in branches mediating the connection between parent and daughter trees.

Abstract

Layering is an important mode of vegetative reproduction at treeline, in which clonal daughter trees are formed by the rooting of lower (“layering”) branches of the parent tree. These branches mediate the connection between parent (PT) and daughter tree (DT). Here, we measured quantity and direction of sap flow in layering branches as well as PT and DT, and measured shoot water potentials in the crowns of a connected PT and DT. We found bidirectional sap flow pattern in layering branches, with the bidirectionality of the flow resulting from water potential dynamics of the parent and daughter trees varying diurnally. We found that 4.3 % of the total water transpired by the DT was supplied by the PT root system, with up to 25 % of the instantaneous daughter tree sap flow coming from the parent tree. In contrast, water provided by the daughter’s root system to the parent tree comprised only a negligible amount, less than 1 % of the parent’s entire sap flow. Additionally, after experimental excavation of part of the DT roots, layering branch flow towards the DT increased, while flows in the opposite direction almost vanished. This study showed that aboveground clonal connections can facilitate a new type of hydraulic redistribution where water is transported bidirectionally through branches. This transfer of water and nutrients is vital especially in the first years of the daughter tree but supplies considerable amounts of water even several years after the establishment of a new clonal tree.

     

Insect visceral muscle. Excitation and conduction in the proctodeal muscles

Bioelectrical potentials were studied from longitudinal muscle fibres of the cockroach proctodeum. The muscle bundle receives a polyaxonal innervation from both anterior and posterior branches of the anterior proctodeal nerve. Evoked post-synaptic potentials consisted of two independent, but similar components generated through the two branches. An action potential in the muscle fibre could be generated with single branch stimulation, and more readily by co-operation of excitation in the two nerve branches.Any part of the muscle was capable of acting as a pacemaker for myogenic rhythmic action potential, and the pacemaker region fluctuated with time. Excitation of the muscle could spread in two ways, directly myogenic and indirectly through nerve tracts. Myogenic conduction (2 cm/sec) was observed to be slower than neural conduction (35–38 cm/sec) in the muscle bundle.     

COMPUTER SIMULATION OF BRANCH INTERACTION AND REGULATION BY UNEQUAL FLOW RATES IN BOTANICAL TREES

Two aspects of branch interaction in trees are investigated theoretically. In the first it is assumed that there is a controlling factor in which the proximity of neighboring terminal branch units influences their branching capability. The almost horizontal tiers of lateral branches of Terminalia catappa L. and Cornus alternifolia L. are simulated by computer using values based on the measured branch geometries of real trees. For branch interaction, we assume a horizontal circle of inhibition whose center is the existing terminal point of a branch. If the end point of another branch extends into the circle, the original branch fails to bifurcate. Examples of computer simulated patterns are illustrated using different degrees of interference and are compared with branch tiers in T. catappa. In the second model the ability of a terminal branch unit to bifurcate is considered to be determined by the accumulation of a critical amount of a hypothetical growth- or bifurcation-determining factor. The daughter branches of a bifurcation are assumed to have differing “flow rates,” i.e., the factor is distributed in different amounts between different daughter axes. Some simulated patterns generated by this model are very similar to real patterns found in T. catappa and an unnamed species of Tabernaemontana. In both simulations bifurcation ratios are determined and are shown to be a variable, not a fixed, property of the simulated trees.     

Multiplicity of pacemaker zones in Helix pomatia neurons

Intracellular microelectrode recordings from neurons ofHelix pomatia revealed several local zones of action potential generation both on the soma and on some of the branches of the neurons. Under certain conditions the activity of individual loci of the neuron membrane was synchronized to produce a normal action potential. It is suggested that the somatic membrane of neurons is heterogeneous in structure and consists of separate loci of an electrically excitable membrane, incorporating active and latent pacemaker zones. Neurons ofH. pomatia are characterized by two types of action potential with different triggering mechanisms: one (synaptic) type is generated under the influence of the EPSP, the other (pacemaker) arises through activation of endogenous factors for the neuron (pacemaker potentials). The interaction between synaptic and pacemaker potentials during integrative activity of the neuron is discussed.M. V. Lomonosov Moscow State University. Translated from Neirofiziologiya, Vol. 5, No. 1, pp. 88–94, January–February, 1973.     

Inflow/Outflow Boundary Conditions for Particle-Based Blood Flow Simulations: Application to Arterial Bifurcations and Trees

When blood flows through a bifurcation, red blood cells (RBCs) travel into side branches at different hematocrit levels, and it is even possible that all RBCs enter into one branch only, leading to a complete separation of plasma and RBCs. To quantify this phenomenon via particle-based mesoscopic simulations, we developed a general framework for open boundary conditions in multiphase flows that is effective even for high hematocrit levels. The inflow at the inlet is duplicated from a fully developed flow generated in a pilot simulation with periodic boundary conditions. The outflow is controlled by adaptive forces to maintain the flow rate and velocity gradient at fixed values, while the particles leaving the arteriole at the outlet are removed from the system. Upon validation of this approach, we performed systematic 3D simulations to study plasma skimming in arterioles of diameters 20 to 32 microns. For a flow rate ratio 6:1 at the branches, we observed the “all-or-nothing” phenomenon with plasma only entering the low flow rate branch. We then simulated blood-plasma separation in arteriolar bifurcations with different bifurcation angles and same diameter of the daughter branches. Our simulations predict a significant increase in RBC flux through the main daughter branch as the bifurcation angle is increased. Finally, we demonstrated the effectiveness of the new methodology in simulations of blood flow in vessels with multiple inlets and outlets, constructed using an angiogenesis model.     

Laser microbeam axotomy and conduction block show that electrical transmission at a central synapse is distributed at multiple contacts

Touch (T) sensory neurons in the leech innervate defined regions of skin and synapse on other neurons, including other T cells, within the ganglionic neuropil. The cells' receptive fields in the periphery are comprised of a central region, innervated by thick axons, and adjoining regions (minor fields) innervated by thinner axons. Secondary branches, known to be sites of synapses, emerge from the thinner and thicker axons. Pairs of T cells appear to make up to 200 separate contacts distributed within the neuropil. When the T cell is hyperpolarized, as occurs during natural stimulation of the cell, action potentials generated in the minor field and travelling into the ganglion along the thin axons may fail to conduct at central branch points. Evidence is presented, using axon conduction block and laser axotomy of cells filled with 6-carboxy-fluorescein, that synapses between separate groups of branches can function independently. Thus, selective activation of branches of the thin anterior axon produced a synaptic potential 36 +/- 6% of control amplitude, which was consistent with counts of 39 +/- 6% of contacts made by these branches. Laser axotomy of postsynaptic neurons showed that the anterior contacts indeed made the principal or only contacts activated during anterior conduction block. The results show that conduction block can modulate transmission within the ganglion, and it operates by silencing particular contacts between cells.     

Influence of amplitude cancellation on the simulated surface electromyogram.

The purpose of the study was to quantify the influence of selected motor unit properties and patterns of activity on amplitude cancellation in the simulated surface electromyogram (EMG). The study involved computer simulations of a motor unit population with physiologically defined recruitment and rate coding characteristics that activated muscle fibers whose potentials were recorded on the skin over the muscle. Amplitude cancellation was quantified as the percent difference in signal amplitude when motor unit potentials were summed before and after rectification. The simulations involved varying the level of activation for the motor unit population, the recording configuration, the upper limit of motor unit recruitment, peak discharge rates, the amount of motor unit synchronization, muscle fiber length, the thickness of the subcutaneous tissue, and the motor unit properties that change with advancing age. The results confirmed a previous experimental report (Day SJ and Hulliger M, J Neurophysiol 86: 2144-2158, 2001) that amplitude cancellation in the surface EMG can reach 62% at maximal activation. A decrease in the range of amplitudes of the motor unit potentials, as can occur during fatiguing contractions, increased amplitude cancellation up to approximately 85%. Differences in the amount of amplitude cancellation were observed across all simulated conditions, and resulted in substantial changes in the absolute magnitude of the EMG signal. The most profound factors influencing amplitude cancellation were the number of active motor units and the duration of the action potentials. The effects of amplitude cancellation were minimal (<5%) when the EMG amplitude was normalized to maximal values, with the exception of variations in peak discharge rate and recruitment range, which resulted in differences up to 17% in the normalized EMG signal across conditions. These results indicate the amount of amplitude cancellation that can occur in various experimental conditions and its influence on absolute and relative measures of EMG amplitude.