Hopping frequency influences elastic energy reuse with joint series elastic actuators.

Robotic limb design struggles to combine energy efficiency with human-like levels of movement versatility. High efficiency and a range of angles and torques are characteristics of human hopping at different frequencies. Humans use muscles in combination with tendons to achieve the required joint actuation. Therefore, we consider whether appropriately tuned series elastic actuators (SEAs) placed at the leg joints can be used to reduce the functional gap between robots and humans. Human hip, knee, and ankle biomechanics were recorded over a range of hopping frequencies to extract joint angles and torques, which were used as an input to a mechanical simulation SEA model. This model was used to optimize the SEA stiffness of each joint to either minimize peak power or energy requirements. This work investigated the relationship between hopping frequency and SEA stiffness, the utility of using SEAs at each joint, and the reasons behind humans' preferred hopping frequency. Although the constant stiffness values across different hopping frequencies are suitable for the knee and the ankle, a variable serial elastic actuator stiffness could still further reduce energy requirements. Optimal SEA stiffness was found to reduce peak power requirements by up to 73% at the ankle and up to 66% at the knee, with greatest benefits found around the preferred frequency. However, no SEA benefits were found for the hip and above the preferred hopping frequency for the knee. These insights could be used to aid in the design of robotic and assistive devices to achieve versatile and energy efficient human-like movements.

Biarticular muscles in light of template models, experiments and robotics: a review.

Leg morphology is an important outcome of evolution. A remarkable morphological leg feature is the existence of biarticular muscles that span adjacent joints. Diverse studies from different fields of research suggest a less coherent understanding of the muscles' functionality in cyclic, sagittal plane locomotion. We structured this review of biarticular muscle function by reflecting biomechanical template models, human experiments and robotic system designs. Within these approaches, we surveyed the contribution of biarticular muscles to the locomotor subfunctions (stance, balance and swing). While mono- and biarticular muscles do not show physiological differences, the reviewed studies provide evidence for complementary and locomotor subfunction-specific contributions of mono- and biarticular muscles. In stance, biarticular muscles coordinate joint movements, improve economy (e.g. by transferring energy) and secure the zig-zag configuration of the leg against joint overextension. These commonly known functions are extended by an explicit role of biarticular muscles in controlling the angular momentum for balance and swing. Human-like leg arrangement and intrinsic (compliant) properties of biarticular structures improve the controllability and energy efficiency of legged robots and assistive devices. Future interdisciplinary research on biarticular muscles should address their role for sensing and control as well as non-cyclic and/or non-sagittal motions, and non-static moment arms.

Gentamicin susceptibility in Escherichia coli related to the genetic background: problems with breakpoints.

In total, 120 Escherichia coli isolates positive for one of the gentamicin resistance (GEN(R)) genes aac(3)-II, aac(3)-IV or ant(2'')-I were tested for gentamicin susceptibility by the agar dilution method. Isolates positive for aac(3)-IV or ant(2'')-I had an MIC distribution of 8-64 mg/L, whereas isolates positive for aac(3)-II had MICs of 32 to >512 mg/L, suggesting a relationship between the distribution of MICs and the specific GEN(R) mechanism. The MIC distribution, regardless of the GEN(R) mechanism, was 8 - >512 mg/L, which supports the clinical breakpoint of MIC >4 mg/L suggested by EUCAST and questions the breakpoint recommended by the CLSI (> or =16 mg/L).

Epidemiology and ecology of enterococci, with special reference to antibiotic resistant strains, in animals, humans and the environment. Example of an ongoing project within the European research programme.

The objectives of the present study are to generate knowledge of the ecology and epidemiology of enterococci in the food chain by studying the following: (1) the population structure (in measures of abundance, number of vancomycin resistant strains, antibiotic resistance patterns, diversity, and stability) among enterococcal populations in different geographical regions and in different links of the food chain (2) possible transmission of strains through the food chain and between hospital environments and the food chain (3) the association between vancomycin resistance and individual strains of enterococci and (4) the diversity of the drug resistance genes in enterococci. So far, 1578 samples have been collected from different countries within the EU (Sweden, Denmark, UK and Spain), and from different habitats (pig farms, carcasses in slaughter houses, soil, manure, water, sewage, and humans). Total and vancomycin resistant enterococcal populations in each sample have been enumerated and more than 12000 isolates have been characterised by phenotyping. Representative isolates are further species identified and characterised by genotyping and MIC determination and from antibiotic resistant isolates the resistance genes are characterised.

Relations between the occurrence of resistance to antimicrobial growth promoters among Enterococcus faecium isolated from broilers and broiler meat.

From 1995 to 2001, Enterococcus faecium isolates were collected from broiler flocks at slaughter and broiler meat products at retail outlets and were tested for susceptibility to classes of antimicrobials used for growth promotion in broilers in Denmark, namely: evernimicin, glycopeptide, macrolide and streptogramin. By February 1998, all antimicrobial growth promoters (AGPs) were withdrawn from the Danish broiler production. The present study investigates, by logistic regression analyses, the (1) changes in the occurrence of AGP resistance among E. faecium from broilers and broiler meat from the fourth quarter of 1995 to the fourth quarter of 2001 and (2) relations between the occurrence of AGP resistance among E. faecium isolates from Danish broilers and AGP resistance among E. faecium isolates from the broiler meat of Danish and unknown origin collected in the same quarter within the year. In the present study, we showed that after the AGP withdrawal, a significant decline in resistance to avilamycin, erythromycin, vancomycin and virginiamycin was observed among E. faecium from broilers and broiler meat. In addition, a decline in the occurrence of AGP resistance among E. faecium from Danish broilers was associated with a decrease in the predicted probability of isolating an AGP-resistant E. faecium isolate from a randomly selected broiler meat product. In the analyses "relations between the occurrence of AGP resistance among E. faecium isolated from broilers and broiler meat collected in the same quarter" errors in the explanatory variable were expected. Therefore, a simulation study was performed to validate the results from logistic regression analyses. The results obtained by the two methods were similar.

Dynamics of the long jump.

A mechanical model is proposed which quantitatively describes the dynamics of the centre of gravity (c.g.) during the take-off phase of the long jump. The model entails a minimal but necessary number of components: a linear leg spring with the ability of lengthening to describe the active peak of the force time curve and a distal mass coupled with nonlinear visco-elastic elements to describe the passive peak. The influence of the positions and velocities of the supported body and the jumper's leg as well as of systemic parameters such as leg stiffness and mass distribution on the jumping distance were investigated. Techniques for optimum operation are identified: (1) There is a minimum stiffness for optimum performance. Further increase of the stiffness does not lead to longer jumps. (2) For any given stiffness there is always an optimum angle of attack. (3) The same distance can be achieved by different techniques. (4) The losses due to deceleration of the supporting leg do not result in reduced jumping distance as this deceleration results in a higher vertical momentum. (5) Thus, increasing the touch-down velocity of the jumper's supporting leg increases jumping distance.

Chromosomal damages by ethanol and acetaldehyde in Saccharomyces cerevisiae as studied by pulsed field gel electrophoresis.

We report on the effect of ethanol and acetaldehyde on yeast chromosomal DNA and on isolated DNA. Ethanol induced DNA single-strand breaks in repair deficient but not in repair proficient Saccharomyces cerevisiae. Acetaldehyde has a deleterious effect on chromosomal DNA in cells as well as on isolated DNA. The results presented support earlier data to show that ethanol is mutagenic via its first metabolite, acetaldehyde.

Multiple-step model-experiment matching allows precise definition of dynamical leg parameters in human running.

The spring-loaded inverted pendulum (SLIP) model is a well established model for describing bouncy gaits like human running. The notion of spring-like leg behavior has led many researchers to compute the corresponding parameters, predominantly stiffness, in various experimental setups and in various ways. However, different methods yield different results, making the comparison between studies difficult. Further, a model simulation with experimentally obtained leg parameters typically results in comparatively large differences between model and experimental center of mass trajectories. Here, we pursue the opposite approach which is calculating model parameters that allow reproduction of an experimental sequence of steps. In addition, to capture energy fluctuations, an extension of the SLIP (ESLIP) is required and presented. The excellent match of the models with the experiment validates the description of human running by the SLIP with the obtained parameters which we hence call dynamical leg parameters.

Integration of intrinsic muscle properties, feed-forward and feedback signals for generating and stabilizing hopping.

It was hypothesized that a tight integration of feed-forward and feedback-driven muscle activation with the characteristic intrinsic muscle properties is a key feature of locomotion in challenging environments. In this simulation study it was investigated whether a combination of feed-forward and feedback signals improves hopping stability compared with those simulations with one individual type of activation. In a reduced one-dimensional hopping model with a Hill-type muscle (one contractile element, neither serial nor parallel elastic elements), the level of detail of the muscle's force-length-velocity relation and the type of activation generation (feed-forward, feedback and combination of both) were varied to test their influence on periodic hopping. The stability of the hopping patterns was evaluated by return map analysis. It was found that the combination of feed-forward and proprioceptive feedback improved hopping stability. Furthermore, the nonlinear Hill-type representation of intrinsic muscle properties led to a faster reduction of perturbations than a linear approximation, independent of the type of activation. The results emphasize the ability of organisms to exploit the stabilizing properties of intrinsic muscle characteristics.

Robust and efficient walking with spring-like legs.

The development of bipedal walking robots is inspired by human walking. A way of implementing walking could be performed by mimicking human leg dynamics. A fundamental model, representing human leg dynamics during walking and running, is the bipedal spring-mass model which is the basis for this paper. The aim of this study is the identification of leg parameters leading to a compromise between robustness and energy efficiency in walking. It is found that, compared to asymmetric walking, symmetric walking with flatter angles of attack reveals such a compromise. With increasing leg stiffness, energy efficiency increases continuously. However, robustness is the maximum at moderate leg stiffness and decreases slightly with increasing stiffness. Hence, an adjustable leg compliance would be preferred, which is adaptable to the environment. If the ground is even, a high leg stiffness leads to energy efficient walking. However, if external perturbations are expected, e.g. when the robot walks on uneven terrain, the leg should be softer and the angle of attack flatter. In the case of underactuated robots with constant physical springs, the leg stiffness should be larger than k = 14 in order to use the most robust gait. Soft legs, however, lack in both robustness and efficiency.

Swing leg control in human running.

Humans can run within a wide range of speeds without thinking about stabilizing strategies. The leg properties seem to be adjusted automatically without need for sensory feedback. In this work, the dynamics of human running are represented by the planar spring mass model. Within this framework, for higher speeds, running patterns can be stable without control strategies. Here, potential strategies that provide stability over a broader range of running patterns are considered and these theoretical predictions are compared to human running data. Periodic running solutions are identified and analyzed with respect to their stability. The control strategies are assumed as linear adaptations of the leg parameters-leg angle, leg stiffness and leg length-during the swing phase. To evaluate the applied control strategies regarding their influence on landing behavior, two parameters are introduced: the velocity of the foot relative to the ground (ground speed matching) and the foot's angle of approach. The results show that periodic running solutions can be stabilized and that control strategies, which guarantee running stability, are redundant. For any swing leg kinematics (adaptation of the leg angle and the leg length), running stability can be achieved by adapting the leg stiffness in anticipation of the ground contact.

The role of intrinsic muscle properties for stable hopping--stability is achieved by the force-velocity relation.

A reductionist approach was presented to investigate which level of detail of the physiological muscle is required for stable locomotion. Periodic movements of a simplified one-dimensional hopping model with a Hill-type muscle (one contractile element, neither serial nor parallel elastic elements) were analyzed. Force-length and force-velocity relations of the muscle were varied in three levels of approximation (constant, linear and Hill-shaped nonlinear) resulting in nine different hopping models of different complexity. Stability of these models was evaluated by return map analysis and the performance by the maximum hopping height. The simplest model (constant force-length and constant force-velocity relations) outperformed all others in the maximum hopping height but was unstable. Stable hopping was achieved with linear and Hill-shaped nonlinear characteristic of the force-velocity relation. The characteristics of the force-length relation marginally influenced hopping stability. The results of this approach indicate that the intrinsic properties of the contractile element are responsible for stabilization of periodic movements. This connotes that (a) complex movements like legged locomotion could benefit from stabilizing effects of muscle properties, and (b) technical systems could benefit from the emerging stability when implementing biological characteristics into artificial muscles.

Upright human gait did not provide a major mechanical challenge for our ancestors.

Habitual bipedalism is considered as a major breakthrough in human evolution and is the defining feature of hominins. Upright posture is presumably less stable than quadrupedal posture, but when using external support, for example, toddlers assisted by their parents, postural stability becomes less critical. In this study, we show that humans seem to mimic such external support by creating a virtual pivot point (VPP) above their centre of mass. A highly reduced conceptual walking model based on this assumption reveals that such virtual support is sufficient for achieving and maintaining postural stability. The VPP is experimentally observed in walking humans and dogs and in running chickens, suggesting that it might be a convenient emergent behaviour of gait mechanics and not an intentional locomotion behaviour. Hence, it is likely that even the first hominis may have already applied the VPP, a mechanism that would have facilitated the development of habitual bipedalism.

Kinetic characterization of plasma membrane ATPase from Saccharomyces cerevisiae.

1. Plasma membrane preparations have been isolated from spheroplasts of Saccharomyces cerevisiae, strain R XII, via lysis and subsequent differential centrifugation. These preparations are almost devoid of mitochondrial contamination. 2. The plasma membrane ATPase is fairly stable when refrigerated, but loses activity at 8 degrees C and above. Below pH 5.6 the ATPase is irreversibly inactivated. The enzyme also splits GTP and ITP, although to a lesser extent. 3. Mg2+-ions are essential as part of the reactive substrate, MgATP, and furthermore they activate the ATPase. Optimal conditions depend on substrate concentration. When the concentration of free Mg2+ ions exceeds about 0.1 mM, competitive inhibition occurs. 4. In the range of pH 5.6-9.2 two functional groups dissociate. One, with pKb = 8.1 +/- 0.1 participated in substrate binding and another one with pKb' = 8.1 +/- 0.1 is involved in substrate splitting. 5. The experiments with group-specific inhibitors suggest that an alpha-amino group and a sulfhydryl residue are involved in substrate binding and conversion. Furthermore, imidazole, tryptophan and carboxyl residues may be important for the catalytic process.

Stable operation of an elastic three-segment leg.

Quasi-elastic operation of joints in multi-segmented systems as they occur in the legs of humans, animals, and robots requires a careful tuning of leg properties and geometry if catastrophic counteracting operation of the joints is to be avoided. A simple three-segment model has been used to investigate the segmental organization of the leg during repulsive tasks like human running and jumping. The effective operation of the muscles crossing the knee and ankle joints is described in terms of rotational springs. The following issues were addressed in this study: (1) how can the joint torques be controlled to result in a spring-like leg operation? (2) how can rotational stiffnesses be adjusted to leg-segment geometry? and (3) to what extend can unequal segment lengths and orientations be advantageous? It was found that: (1) the three-segment leg tends to become unstable at a certain amount of bending expressed by a counterrotation of the joints; (2) homogeneous bending requires adaptation of the rotational stiffnesses to the outer segment lengths; (3) nonlinear joint torque-displacement behaviour extends the range of stable leg bending and may result in an almost constant leg stiffness; (4) biarticular structures (like human gastrocnemius muscle) and geometrical constraints (like heel strike) support homogeneous bending in both joints; (5) unequal segment lengths enable homogeneous bending if asymmetric nominal angles meet the asymmetry in leg geometry; and (6) a short foot supports the elastic control of almost stretched knee positions. Furthermore, general leg design strategies for animals and robots are discussed with respect to the range of safe leg operation.

Antimicrobial resistance in Salmonella enterica subsp. enterica serovar typhimurium from humans and production animals.

We have studied the frequency of antimicrobial resistance and epidemiological relatedness among 473 isolates of Salmonella enterica subsp. enterica serovar typhimurium (S. typhimurium) from human and veterinary sources. The human strains were clinical isolates from patients with diarrhoea sent to the State Serum Institute during August 1993 (228 isolates). The animal strains were isolated from clinical or subclinical infections in cattle (48 isolates), pigs (99 isolates) or poultry (98 isolates), all from 1993. All strains were tested against 22 different antimicrobial agents used in both human and veterinary medicine with the tablet diffusion method. Strains were also phage-typed and the plasmid content determined in all resistant strains. Ribotyping was performed on selected strains. Of 228 human isolates tested, 19.3% of the strains were resistant to one or more antimicrobial agent compared with 10.4% of strains from cattle, 11.1% of strains from pigs and 9.2% of strains from poultry. Multiple resistance, i.e. resistance against at least four antimicrobial agents, was found in 9.2% of the human strains, but in only two of the cattle isolates. The majority of the multi-resistant strains in humans were from infections contracted outside Denmark, most often in southern Europe or south-east Asia. Resistance in human strains was most common against tetracycline (13%), ampicillin (12%), sulphonamide (12%), streptomycin (10%) and chloramphenicol (8%). The resistance pattern differed somewhat in animal isolates: Poultry strains were usually resistant only to ampicillin, while pig and cattle isolates were most often resistant to sulphonamide, tetracycline and streptomycin. Typing of the strains showed that some animal strains and human strains were indistinguishable. In conclusion, while antimicrobial resistance was present in S. typhimurium isolated from humans and animals in Denmark, multiple resistance was most often acquired outside Denmark.

Optimum take-off techniques and muscle design for long jump.

A two-segment model based on Alexander (1990; Phil. Trans. R. Soc. Lond. B 329, 3-10) was used to investigate the action of knee extensor muscles during long jumps. A more realistic representation of the muscle and tendon properties than implemented previously was necessary to demonstrate the advantages of eccentric force enhancement and non-linear tendon properties. During the take-off phase of the long jump, highly stretched leg extensor muscles are able to generate the required vertical momentum. Thereby, serially arranged elastic structures may increase the duration of muscle lengthening and dissipative operation, resulting in an enhanced force generation of the muscle-tendon complex. To obtain maximum performance, athletes run at maximum speed and have a net loss in mechanical energy during the take-off phase. The positive work done by the concentrically operating muscle is clearly less than the work done by the surrounding system on the muscle during the eccentric phase. Jumping performance was insensitive to changes in tendon compliance and muscle speed, but was greatly influenced by muscle strength and eccentric force enhancement. In agreement with a variety of experimental jumping performances, the optimal jumping technique (angle of attack) was insensitive to the approach speed and to muscle properties (muscle mass, the ratio of muscle fibre to tendon cross-sectional area, relative length of fibres and tendon). The muscle properties also restrict the predicted range of the angle of the velocity vector at take-off.

Relations between the consumption of antimicrobial growth promoters and the occurrence of resistance among Enterococcus faecium isolated from broilers.

The present study investigates, at farm level, the effect of the time-span between sampling and the last time a particular antimicrobial growth promoter (AGP) was included in the feed on the probability of selecting an AGP-resistant Enterococcus faecium isolate from a broiler flock. The probability that a randomly selected E. faecium isolate was resistant to avilamycin, erythromycin or virginiamycin was 0.91, 0.92 and 0.84, respectively if the isolate originated from a broiler flock fed either avilamycin- or virginiamycin-supplemented feed. As the time-span between sampling and the last AGP consumption increased, the probability of isolating an E. faecium isolate resistant to a particular AGP decreased (probability <0.2 within 3-5 years after last exposure to AGPs). The decrease in probability over time showed little farm-to-farm variation. The number of times a particular AGP was given to previous flocks reared in the same house had no effect on the probability of isolating a resistant isolate.

Effects of a heterogeneous set of xenobiotics on growth and plasma membranes of mammalian and fungal cell cultures.

A comparison of the toxicity of 45 selected, heterogenous substances on two test organisms of different taxonomic levels, the yeast Saccharomyces cerevisiae and Chinese hamster ovary (CHO) cells, was made. In addition, effects on the yeast plasma membrane-integrated H(+)-ATPase and on the CHO adenosine uptake system were investigated. For all test systems, log EC50 values highly correlated with log EC20 values. Good correlations were obtained between CHO proliferation rate and yeast growth rate (r = 0.80). However, CHO cells were about four times more sensitive than yeast. A good accordance was also found between effects on yeast cell growth and on the H(+)-ATPase, indicating a plasma membrane impairment as a major cause of cytotoxicity. These findings were supported by correlations of log EC20 values with the log Pow as a measure for lipophilicity. Although the test systems demonstrated different dependencies, the main trend reflected an increasing toxicity with increasing lipophilicity. Comparisons with data from in vivo test systems suggest that these in vitro test systems could be implemented for initial estimation of basic toxicity and the detection of outliers thereby reducing the number of tests with higher animals.