The load dependence of kinesin’s mechanical cycle

Kinesin is a dimeric motor protein that transports organelles in a stepwise manner toward the plus-end of microtubules by converting the energy of ATP hydrolysis into mechanical work. External forces can influence the behavior of kinesin, and force-velocity curves have shown that the motor will slow down and eventually stall under opposing loads of ≈5 pN. Using an in vitro motility assay in conjunction with a high-resolution optical trapping microscope, we have examined the behavior of individual kinesin molecules under two previously unexplored loading regimes: super-stall loads (>5 pN) and forward (plus-end directed) loads. Whereas some theories of kinesin function predict a reversal of directionality under high loads, we found that kinesin does not walk backwards under loads of up to 13 pN, probably because of an irreversible transition in the mechanical cycle. We also found that this cycle can be significantly accelerated by forward loads under a wide range of ATP concentrations. Finally, we noted an increase in kinesin’s rate of dissociation from the microtubule with increasing load, which is consistent with a load dependent partitioning between two recently described kinetic pathways: a coordinated-head pathway (which leads to stepping) and an independent-head pathway (which is static).

Unbinding process of adsorbed proteins under external stress studied by atomic force microscopy spectroscopy

We report the study of the dynamics of the unbinding process under a force load f of adsorbed proteins (fibrinogen) on a solid surface (hydrophilic silica) by means of atomic force microscopy spectroscopy. By varying the loading rate rf, defined by f = rf t, t being the time, we find that, as for specific interactions, the mean rupture force increases with rf. This unbinding process is analyzed in the framework of the widely used Bell model. The typical dissociation rate at zero force entering in the model lies between 0.02 and 0.6 s−1. Each measured rupture is characterized by a force f0, which appears to be quantized in integer multiples of 180–200 pN.

External perturbation of the trunk in standing humans differentially activates components of the medial back muscles

During voluntary arm movements, the medial back muscles are differentially active. It is not known whether differential activity also occurs when the trunk is perturbed unpredictably, when the earliest responses are initiated by short-latency spinal mechanisms rather than voluntary commands. To assess this, in unpredictable and self-initiated conditions, a weight was dropped into a bucket that was held by the standing subject (n = 7). EMG activity was recorded from the deep (Deep MF), superficial (Sup MF) and lateral (Lat MF) lumbar multifidus, the thoracic erector spinae (ES) and the biceps brachii. With unpredictable perturbations, EMG activity was first noted in the biceps brachii, then the thoracic ES, followed synchronously in the components of the multifidus. During self-initiated perturbations, background EMG in the Deep MF increased two- to threefold, and the latency of the loading response decreased in six out of the seven subjects. In Sup MF and Lat MF, this increase in background EMG was not observed, and the latency of the loading response was increased. Short-latency reflex mechanisms do not cause differential action of the medial back muscles when the trunk is loaded. However, during voluntary tasks the central nervous system exerts a 'tuned response', which involves discrete activity in the deep and superficial components of the medial lumbar muscles in a way that varies according to the biomechanical action of the muscle component.

The effects of varying time under tension and volume load on acute neuromuscular responses

The purpose of this study was to examine the effects of different methods of measuring training volume, controlled in different ways, on selected variables that reflect acute neuromuscular responses. Eighteen resistance-trained males performed three fatiguing protocols of dynamic constant external resistance exercise, involving elbow flexors, that manipulated either time-under-tension (TUT) or volume load (VL), defined as the product of training load and repetitions. Protocol A provided a standard for TUT and VL. Protocol B involved the same VL as Protocol A but only 40% concentric TUT; Protocol C was equated to Protocol A for TUT but only involved 50% VL. Fatigue was assessed by changes in maximum voluntary isometric contraction (MVIC), interpolated doublet (ID), muscle twitch characteristics (peak twitch, time to peak twitch, 0.5 relaxation time, and mean rates of force development and twitch relaxation). All protocols produced significant changes (P

Assessments of training load in elite youth soccer

One of the most popular sports globally, soccer has seen a rise in the demands of the game over recent years. An increase in intensity and playing demands, coupled with growing social and economic pressures on soccer players means that optimal preparation is of paramount importance. Recent research has found the modern game, depending on positional role, to consist of approximately 60% more sprint distance in the English Premier League, which was also found to be the case for frequency and success of discrete technical actions (Bush et al., 2015). As a result, the focus on soccer training and player preparedness is becoming more prevalent in scientific research. By designing the appropriate training load, and thus periodization strategies, the aim is to achieve peak fitness in the most efficient way, whilst minimising the risk of injury and illness. Traditionally, training intensity has been based on heart rate responses, however, the emergence of tracking microtechnology such as global positioning system (GPS) and inertial sensors are now able to further quantify biomechanical load as well as physiological stress. Detailed pictures of internal and external loading indices such as these then combine to produce a more holistic view of training load experience by the player during typical drills and phases of training in soccer. The premise of this research is to gain greater understanding of the physical demands of common training methodologies in elite soccer to support optimal match performance. The coaching process may then benefit from being able to prescribe the most effective training to support these. The first experimental chapter in this thesis began by quantify gross training loads of the pre-season and in-season phases in soccer. A broader picture of the training loads inherent in these distinct phases brought more detail as to the type and extent of external loading experienced by soccer players at these times, and how the inclusion of match play influences weekly training rhythms. Training volume (total distance) was found to be high at the start compared to the end of pre-season (37 kilometres and 28 kilometres), where high cardiovascular loads were attained as part of the conditioning focus. This progressed transiently, however, to involve higher-speed, acceleration and change-of-direction stimuli at the end of pre-season compared to the start and to that in-season (1.18 kilometres, 0.70 kilometres and 0.42 kilometres high-intensity running; with 37, 25 and 23 accelerations >3m/s2 respectively) . The decrease in volume and increase in maximal anaerobic activity was evident in the training focus as friendly matches were introduced before the competitive season. The influence of match-play as being a large physical dose in the training week may then determine the change in weekly periodisation and how resulting training loads applied and tapered, if necessary. The focus of research was then directed more specifically to the most common mode of training in soccer, that also featured regularly in the pre-season period in the present study, small-sided games (SSG). The subsequent studies examined numerous manipulations of this specific form of soccer conditioning, such as player numbers as well as absolute and relative playing space available. In contrast to some previous literature, changing the number of players did not seem to influence training responses significantly, although playing format in the possession style brought about larger effects for heart rate (89.9%HRmax) and average velocity (7.6km/h-1). However, the following studies (Chapters 5, 6 and 7) revealed a greater influence of relative playing space available to players in SSG. The larger area at their disposal brought about greater aerobic responses (~90%HRmax), by allowing higher average and peak velocities (>25km/h-1), as well as greater distance acceleration behaviour at greater thresholds (>2.8m/s2). Furthermore, the data points towards space as being a large determinant in strategy of the player in small-sided games (SSG), subsequently shaping their movement behaviour and resulting physical responses. For example, higher average velocities in a possession format (8km/h-1) reflects higher work rate and heart rate load but makes achieving significant neuromuscular accelerations at a high level difficult given higher starting velocities prior to the most intense accelerations (4.2km/h-1). By altering space available and even through intentional numerical imbalances in team numbers, it may be easier for coaches to achieve the desired stimulus for the session or individual player, whether that is for aerobic and neuromuscular conditioning. Large effects were found for heart rate being higher in the underloaded team (85-90%HRmax) compared to the team with more players (80-85%HRmax) as well as for RPE (5AU versus 7AU). This was also apparent for meterage and therefore average velocity. It would also seem neuromuscular load through high acceleration and deceleration efforts were more pronounced with less numbers (given the need to press and close down opponents, and in a larger area relative to the number of players on the underloaded team. The peak accelerations and deceleration achieved was also higher when playing with less players (3-6.2m/s2 and 3-6.1m/s2) Having detailed ways in which to reach desired physical loading responses in common small training formats, Chapter 8 compared SSG to larger 9v9 formats with full-size 11v11 friendly matches. This enabled absolute and relative comparisons to be made and to understand the extent to which smaller training formats are able to replicate the required movements to be successful in competition. In relative terms, it was revealed that relative acceleration distance and Player Load were higher in smaller 4v4 games than match-play (1.1m.min-1 and 0.3m.min-1 >3m/s2; 16.9AU versus 12AU). Although the smallest format did not replicate the high-velocity demands of matches, the results confirmed their efficacy in providing significant neuromuscular load during the training week, which may then be supplemented by high-intensity interval running in order to gain exposure to more maximal speed work. In summary, the data presented provide valuable information from GPS and inertial sensor microtechnology which may then be used to understand training better to manipulate types of load according to physical conditioning objectives. For example, a library of resources to direct planning of drills of varying cardiovascular, neuromuscular and perceptual load can be created to give more confidence in session outcomes. Combining external and internal load data of common soccer training drills, and their application across different phases and training objectives may give coaches a powerful tool to plan and periodize training.

Implication of global warming on air-conditioned office buildings in Australia

With the accelerated trend of global warming, the thermal behavior of existing buildings, which were typically designed based on current weather data, may not be able to cope with the future climate. This paper quantifies, through computer simulations, the increased cooling loads imposed by potential global warming and probable indoor temperature increases due to possible undersized air-conditioning system. It is found from the sample office building examined that the existing buildings would generally be able to adapt to the increasing warmth of 2030 year Low and High scenarios projections and 2070 year Low scenario projection. However, for the 2070 year High scenario, the study indicates that the existing office buildings, in all capital cities except for Hobart, will suffer from overheating problems. When the annual average temperature increase exceeds 2°C, the risk of current office buildings subjected to overheating will be significantly increased. For existing buildings which are designed with current climate condition, it is shown that there is a nearly linear correlation between the increase of average external air temperature and the increase of building cooling load. For the new buildings, in which the possible global warming has been taken into account in the design, a 28-59% increase of cooling capacity under 2070 High scenario would be required to improve the building thermal comfort level to an acceptable standard.

A Method of Preparing Future Hourly Weather Data for the Study of Global Warming Impact on the Built Environment

The dynamic interaction between building systems and external climate is extremely complex, involving a large number of difficult-to-predict variables. In order to study the impact of global warming on the built environment, the use of building simulation techniques together with forecast weather data are often necessary. Since all building simulation programs require hourly meteorological input data for their thermal comfort and energy evaluation, the provision of suitable weather data becomes critical. Based on a review of the existing weather data generation models, this paper presents an effective method to generate approximate future hourly weather data suitable for the study of the impact of global warming. Depending on the level of information available for the prediction of future weather condition, it is shown that either the method of retaining to current level, constant offset method or diurnal modelling method may be used to generate the future hourly variation of an individual weather parameter. An example of the application of this method to the different global warming scenarios in Australia is presented. Since there is no reliable projection of possible change in air humidity, solar radiation or wind characters, as a first approximation, these parameters have been assumed to remain at the current level. A sensitivity test of their impact on the building energy performance shows that there is generally a good linear relationship between building cooling load and the changes of weather variables of solar radiation, relative humidity or wind speed.