The effect of mechanical stimulation on root and shoot development of young containerised Quercus robur and Robinia pseudoacacia trees

Thigmomorphogenesis is a well-studied process in agricultural crops and coniferous trees. Nevertheless, the effects on both shoot and root characteristics for deciduous woody species received little attention so far. In this study, the objective was to understand the effect of aboveground flexing treatments on the development of structural, mechanical and physiological root and shoot characteristics for two deciduous tree species, Black locust (Robinia pseudoacacia L.) and English oak (Quercus robur L.). Flexing treatments were performed using an electromechanical device with a rotating arm touching and bending the plants at regular intervals. A wide range of stem, shoot as well as root system characteristics was measured. The different flexing treatments altered above- and belowground plant development for both species, with strongest effects on Quercus and most significant differences between the control and the unidirectional flexing treatment. Some responses are in accordance with previous findings, such as stem eccentricity and reduced shoot elongation under unidirectional flexing, but others are renewing, such as the lower stomatal density and larger epidermal cell surface for the Quercus plants under variable flexing direction. Despite some common responses, both species frequently differed in the way they were affected. Belowground, Quercus plants under unidirectional flexing invested relatively more in their first order root and deeper second order roots, whereas Robinia plants allocated relatively more to fine root biomass and horizontal shallow roots. Both strategies potentially increased pull-out as well as overturning resistance in their own way. The presented findings are valid for young trees grown in small containers. Based on practical know-how and shortcomings experienced in the course of this experiment, methodological recommendations are formulated. We finally stress the complex variability in growth responses, especially for root systems, observed in different studies and related to dissimilarity in species, soil conditions, plant history or type of mechanical perturbation.     

Motility and chemotaxis of Spirochaeta aurantia: computer-assisted motion analysis.

A computer program has been designed to study behavior in populations of Spirochaeta aurantia cells, and this program has been used to analyze changes in behavior in response to chemoattractants. Three kinds of behavior were distinguished: smooth swimming, flexing, and reversals in direction of swimming after a short pause (120 ms). Cell populations exposed to chemoattractants spent, on average, 66, 33, and 1% of the time in these modes, respectively. After the addition of a chemoattractant, behavior was modified transiently--smooth swimming increased, flexing decreased, and reversals were suppressed. After addition of D-xylose (final concentration, 10 mM), the adaptation time (the time required for the populations to return to the unmodified behavior) for S. aurantia was 1.5 to 2.0 min. A model to explain the behavior of S. aurantia and the response of cells to chemoattractants is described. This model includes a coordinating mechanism for flagellar motor operation and a motor switch synchronizing device.     

The effects of air flow and stem flexure on the mechanical and hydraulic properties of the stems of sunflowers Helianthus annuus L

Many studies have shown that wind affects plant development, causing them to develop shorter and usually stronger stems. Many of these effects have been shown to be due to a response to mechanical flexing of the stem which is known as thigmomorphogenesis. However, it is not known how wind affects the hydraulic properties of stems, nor have the effects of air flow past leaves been examined in isolation from mechanical flexing. This study, therefore, used a factorial experiment to distinguish between the effects of stem flexing and air flow, and examined the morphology, hydraulics and mechanics of developing sunflowers Helianthus annuus. It was found that flexure and air flow had opposite effects on several aspects of development; air flow increased plant height and length-specific stem hydraulic conductivity, k(h), and reduced stem rigidity and strength, while flexing did the reverse. There was also a clear trade-off between hydraulic and mechanical capability: as one increased the other decreased. A plant's response to wind must, therefore, be a complex response to at least two different stimuli and this might help explain why it varies with species and environment.     

Movement and distribution of adult rusty grain beetle, Cryptolestes ferrugineus (Coleoptera: Laemophloeidae), in stored wheat in response to different temperature gradients and insect densities

The movement and distribution of adult Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae) in grain provide important information for detection of insect pests and for simulations of their distribution in grain bins. Adult movement and distribution were determined in 100 by 100 by 1000-mm wheat (14.5 +/- 0.2% moisture content) columns at four insect densities, three temperature gradients, and dynamic (changing) temperature conditions. Insect density was a minor factor influencing insect movement and distribution in grain columns with temperature gradients. Dispersal resulted in a uniform distribution at a higher insect density (higher than two adults per kilogram of wheat), and aggregation occurred at a low insect density. Adults wandered in the first 6 h after introduction, and there were fewer adults wandering in the vertical direction than in the horizontal direction. Adults moved faster in the vertical direction than in the horizontal direction, and the maximum speed of the movement was 6 m/d in the horizontal direction, and >10.8 m/d in the vertical direction through wheat. Adults could detect temperature gradients in <1 h and preferred warmer temperatures when they had a choice. Insect distribution in horizontal wheat columns at any temperature gradient was unstable for 24 h. Twenty-four hours after introduction, adults gradually overcame their positive geotactic behavior if the upper temperature was more biologically suitable or was not <27.5 degrees C. Adults responded faster to higher temperature gradients than to lower temperature gradients. There was a similar pattern of adult distribution in 144 h.     

Analysis of estuarine soil gradients in salt marshes of the southwestern Netherlands with special reference to the Scheldt estuary

Macro- and microgradients in salt-marsh soils of the Scheldt estuary have been studied. The soil parameters used are clay, carbonate, organic matter, pH, nitrogen, potassium and phosphate. The hypothesis that estuarine soil gradients can be reflections of similar aquatic gradients is affirmed for two gradients. Soil gradients of upstream increasing organic matter and phosphate contents have been found, relating significantly to corresponding aquatic gradients caused by river pollution. In the same direction gradients of decreasing nitrogen and potassium contents occur. A macrogradient in carbonate content was not found. Some microgradients in clay and carbonate contents are described, and relevant mutual relationships between the soil parameters are characterized. Communication nr. 149 of the Delta Institute for Hydrobiological Research, Yerseke, The Netherlands     

Activation of the Shoulder-Belt and Shoulder Muscles in Two-Joint Arm Movements Performed in Humans with the Action of Opposite Loadings

In tests on four volunteers, we examined coordination of central motor commands (CMCs) controlling slow two-joint movements of the arm within the horizontal plane. Current amplitudes of EMGs recorded from six muscles of the shoulder belt and shoulder and subjected to full-wave rectifying and low-frequency filtration were considered correlates of these commands. In particular, we studied the dependence of coordination of CMCs on the direction of an external force applied to the distal forearm part. As was found, coordination of CMCs significantly depends on the direction of the force flexing the elbow joint. According to our observations, EMGs of definite muscles in the case of performance of a two-joint movement can, in a first approximation, be presented as linear combinations of the EMGs recorded in the course of separate sequential single-joint movements under conditions of shifting the reference point of the hand toward the same point of the operational space as that in the two-joint movement. These data can be interpreted as confirmation of the principle of superposition of elementary CMCs in the performance of complex movements of the extremity.     

Preserved ligament in the Jurassic bivalve Lithiotis: adaptive and evolutionary significance

An exceptionally well-preserved specimen of Lithiotis problematica from the Lower Jurassic of NE Italy possesses multiple resilia, composed of thin fibres and joining both valves, housed in ligamental grooves. The resilia were modified to allow small changes in the reciprocal distance of the valves. This increased the area in which the extremely thin free valve flexed to allow shell closure, and reduced the risk of breakage by fatigue of the valve and/or ligament. The fact that the valves remained connected by ligament for an extended height of the ligamental area implies that (1) the free valve closed by flexing, not by articulating, (2) it did not migrate in the ventral direction as often observed in cemented bivalves, and (3) it was approximately as long as the attached one.     

Microvascular Endothelial Cells Migrate Upstream and Align Against the Shear Stress Field Created by Impinging Flow

At present, little is known about how endothelial cells respond to spatial variations in fluid shear stress such as those that occur locally during embryonic development, at heart valve leaflets, and at sites of aneurysm formation. We built an impinging flow device that exposes endothelial cells to gradients of shear stress. Using this device, we investigated the response of microvascular endothelial cells to shear-stress gradients that ranged from 0 to a peak shear stress of 9–210 dyn/cm². We observe that at high confluency, these cells migrate against the direction of fluid flow and concentrate in the region of maximum wall shear stress, whereas low-density microvascular endothelial cells that lack cell-cell contacts migrate in the flow direction. In addition, the cells align parallel to the flow at low wall shear stresses but orient perpendicularly to the flow direction above a critical threshold in local wall shear stress. Our observations suggest that endothelial cells are exquisitely sensitive to both magnitude and spatial gradients in wall shear stress. The impinging flow device provides a, to our knowledge, novel means to study endothelial cell migration and polarization in response to gradients in physical forces such as wall shear stress.     

Microvascular Endothelial Cells Migrate Upstream and Align Against the Shear Stress Field Created by Impinging Flow

At present, little is known about how endothelial cells respond to spatial variations in fluid shear stress such as those that occur locally during embryonic development, at heart valve leaflets, and at sites of aneurysm formation. We built an impinging flow device that exposes endothelial cells to gradients of shear stress. Using this device, we investigated the response of microvascular endothelial cells to shear-stress gradients that ranged from 0 to a peak shear stress of 9–210 dyn/cm². We observe that at high confluency, these cells migrate against the direction of fluid flow and concentrate in the region of maximum wall shear stress, whereas low-density microvascular endothelial cells that lack cell-cell contacts migrate in the flow direction. In addition, the cells align parallel to the flow at low wall shear stresses but orient perpendicularly to the flow direction above a critical threshold in local wall shear stress. Our observations suggest that endothelial cells are exquisitely sensitive to both magnitude and spatial gradients in wall shear stress. The impinging flow device provides a, to our knowledge, novel means to study endothelial cell migration and polarization in response to gradients in physical forces such as wall shear stress.     

Cells as Active Particles in Asymmetric Potentials: Motility under External Gradients

Cell migration is a crucial event during development and in disease. Mechanical constraints and chemical gradients can contribute to the establishment of cell direction, but their respective roles remain poorly understood. Using a microfabricated topographical ratchet, we show that the nucleus dictates the direction of cell movement through mechanical guidance by its environment. We demonstrate that this direction can be tuned by combining the topographical ratchet with a biochemical gradient of fibronectin adhesion. We report competition and cooperation between the two external cues. We also quantitatively compare the measurements associated with the trajectory of a model that treats cells as fluctuating particles trapped in a periodic asymmetric potential. We show that the cell nucleus contributes to the strength of the trap, whereas cell protrusions guided by the adhesive gradients add a constant tunable bias to the direction of cell motion.     

Cells as Active Particles in Asymmetric Potentials: Motility under External Gradients

Cell migration is a crucial event during development and in disease. Mechanical constraints and chemical gradients can contribute to the establishment of cell direction, but their respective roles remain poorly understood. Using a microfabricated topographical ratchet, we show that the nucleus dictates the direction of cell movement through mechanical guidance by its environment. We demonstrate that this direction can be tuned by combining the topographical ratchet with a biochemical gradient of fibronectin adhesion. We report competition and cooperation between the two external cues. We also quantitatively compare the measurements associated with the trajectory of a model that treats cells as fluctuating particles trapped in a periodic asymmetric potential. We show that the cell nucleus contributes to the strength of the trap, whereas cell protrusions guided by the adhesive gradients add a constant tunable bias to the direction of cell motion.     

Using light to shape chemical gradients for parallel and automated analysis of chemotaxis

Numerous molecular components have been identified that regulate the directed migration of eukaryotic cells toward sources of chemoattractant. However, how the components of this system are wired together to coordinate multiple aspects of the response, such as directionality, speed, and sensitivity to stimulus, remains poorly understood. Here we developed a method to shape chemoattractant gradients optically and analyze cellular chemotaxis responses of hundreds of living cells per well in 96‐well format by measuring speed changes and directional accuracy. We then systematically characterized migration and chemotaxis phenotypes for 285 siRNA perturbations. A key finding was that the G‐protein G_iα subunit selectively controls the direction of migration while the receptor and Gβ subunit proportionally control both speed and direction. Furthermore, we demonstrate that neutrophils chemotax persistently in response to gradients of fMLF but only transiently in response to gradients of ATP. The method we introduce is applicable for diverse chemical cues and systematic perturbations, can be used to measure multiple cell migration and signaling parameters, and is compatible with low‐ and high‐resolution fluorescence microscopy.     

The internal phosphodiesterase RegA is essential for the suppression of lateral pseudopods during Dictyostelium chemotaxis

Dictyostelium strains in which the gene encoding the cytoplasmic cAMP phosphodiesterase RegA is inactivated form small aggregates. This defect was corrected by introducing copies of the wild-type regA gene, indicating that the defect was solely the consequence of the loss of the phosphodiesterase. Using a computer-assisted motion analysis system, regA(-) mutant cells were found to show little sense of direction during aggregation. When labeled wild-type cells were followed in a field of aggregating regA(-) cells, they also failed to move in an orderly direction, indicating that signaling was impaired in mutant cell cultures. However, when labeled regA(-) cells were followed in a field of aggregating wild-type cells, they again failed to move in an orderly manner, primarily in the deduced fronts of waves, indicating that the chemotactic response was also impaired. Since wild-type cells must assess both the increasing spatial gradient and the increasing temporal gradient of cAMP in the front of a natural wave, the behavior of regA(-) cells was motion analyzed first in simulated temporal waves in the absence of spatial gradients and then was analyzed in spatial gradients in the absence of temporal waves. Our results demonstrate that RegA is involved neither in assessing the direction of a spatial gradient of cAMP nor in distinguishing between increasing and decreasing temporal gradients of cAMP. However, RegA is essential for specifically suppressing lateral pseudopod formation during the response to an increasing temporal gradient of cAMP, a necessary component of natural chemotaxis. We discuss the possibility that RegA functions in a network that regulates myosin phosphorylation by controlling internal cAMP levels, and, in support of that hypothesis, we demonstrate that myosin II does not localize in a normal manner to the cortex of regA(-) cells in an increasing temporal gradient of cAMP.     

Flagellation of Pseudomonas putida and analysis of its motile behavior.

Pseudomonas putida flagella were examined. Also, changes in motile behavior in response to chemoattractants were analyzed quantitatively by computer. Reversals in the rotation direction of bundles of polar flagella resulted in changes in swimming direction. Cells swimming in buffer changed direction once every 2 s on average, whereas cells exposed to the attractant benzoate changed direction an average of once every 10 s. The findings show that P. putida responds to temporal gradients of chemoattractant by suppressing changes in the direction of rotation of flagella.     

Isokinetic plantar flexion: experimental results and model calculations

In isokinetic experiments on human subjects, conducted to determine moments that can be exerted about a joint at different angular velocities, joint rotation starts as soon as the moment increases above the resting level. This contraction history differs from the one in experiments on isolated muscle, where the force is allowed to increase to an isometric level before shortening is initiated. The purpose of the present study was to determine the influence of contraction history on plantar flexing moments found during maximal voluntary plantar flexion on an isokinetic dynamometer. In ten subjects, plantar flexing moments were measured as a function of ankle angle at different angular velocities. They were also calculated using a model of the muscle-tendon complex of the human triceps surae. The model incorporates elastic tendinous tissue in series with muscle fibers. The input of the model consists of time histories of active state (the force generating capacity of contractile elements) and shortening velocity of the muscle-tendon complex. Different time courses of active state were offered at fixed length of the muscle-tendon complex. The time course yielding a close match between the calculated rise of plantar flexing moment and the rise measured during fixed angle contractions was used to calculate moment-angle curves for isokinetic plantar flexion. The active state value reached when a peak occurred in calculated moment-angle curves was found to be lower if the angular velocity was made higher. Comparing measured and calculated results, it was concluded that moment-angular velocity diagrams determined in studies of isokinetic plantar flexion in human subjects reflect not only the influence of shortening velocity of contractile elements on the force which can be produced by plantar flexors.     

A model of the human triceps surae muscle-tendon complex applied to jumping

The purpose of this study was to gain more insight into the behavior of the muscle-tendon complex of human m. triceps surae in jumping. During one-legged vertical jumps of ten subjects ground reaction forces as well as cinematographic data were registered, and electromyograms were recorded from m. soleus and m. gastrocnemius. A model was developed of m. triceps surae, incorporating assumptions concerning dimensions, architecture, force-length and force-velocity relationships of muscle fibers, as well as assumptions concerning dimensions and elastic behavior of tendinous tissue in series with the muscle fibers. The velocity with which origin approaches insertion (V OI) was calculated for m. soleus and m. gastrocnemius using cine film data, and served as input of the model. During the last part of the push-off phase EMG-levels were found to be more or less constant, V OI of m. soleus and m. gastrocnemius rapidly increased, and the plantar flexing moment obtained by solving equations concerning a free body diagram of the foot rapidly declined. A similar decline was observed in the plantar flexing moment obtained by multiplying force calculated with help of the model by estimated moment arm at the ankle. As a result of the decline of exerted force tendon length decreases. According to the model the shortening velocity of tendon reaches higher values than that of muscle fibers. The results of a kinetic analysis demonstrate that during the last part of the push-off phase a combination of high angular velocities with relatively large plantar flexing moments is required. It is concluded that without a compliant tendon m. triceps surae would not be able to satisfy this requirement.     

Cortical bone thickness can adapt locally to muscular loading while changing with age

Mechanical loading of muscle action is concentrated at muscle attachment sites; thus there may be a potential for site-specific variation in cortical bone thickness. Humeri from an early 20th-century Finnish (Helsinki) and two medieval English (Newcastle, Blackgate and York, Barbican) populations were subjected to pQCT scanning to calculate site-specific cross-sectional cortical bone area (CA) for four locations and to measure cortical thickness at muscle attachment sites and non-attachment sites. We found that CA at 80% of humerus length was significantly reduced compared to more distal cross-sections, which can be due to reduced stresses at the proximal shaft. The principal direction of loading at 80% humerus length was towards mediolateral plane, likely due to fixing the humerus close to the torso. At 35% the main direction of loading was towards anteroposterior plane, reflecting elbow flexing forces. The principal direction of loading varied between populations, sides and sexes at 50% humerus length due to preference between elbow and shoulder joint; thus this location might be useful when trying to infer differences in activity. These changes are likely due to overall shaft adaptation to forces acting at the humerus. In addition, we found a potential for site-specific variation in cortical thickness; cortical bone at muscle attachment sites was significantly thicker compared to non-attachment sites. Lastly, CA at 35% of humerus length and cortical thickness at non-attachment sites decreased with age. These results underline the importance of muscle loading for bone mass preservation as well as indicate that a site-specific variation of bone mass is possible.     

Evidence for intraspecific phenotypic variation in songbirds along elevation gradients in central Europe

Studying phenotypic variations along gradients may provide insights into mechanisms that drive species distributions and thus can be useful indicators of environmental change. In mountains, the study of phenotypic variation along elevation gradients is of increasing relevance due to the impacts of climate change. We analysed European ringing data to measure the direction of phenotypic variation along elevation gradients in six common, resident songbird species occurring along a wide elevational range. We modelled intraspecific change in wing length, body mass and their ratio with elevation and found a significant increase in wing length and a decrease in body mass at high elevations. The results of our exploratory analysis show the potential that continent-wide ringing databases offer to describe patterns of phenotypic variation along environmental gradients.     

Arrestment responses of the predatory mite, Phytoseiulus persimilis, to steep odour gradients of a kairomone

ABSTRACT. The response of the predatory mite, Phytoseiulus persimilis Athias-Henriot, to steep gradients of a volatile kairomone emitted by its prey, Tetranychus urticue Koch, was studied in a vertical air flow chamber. The orientation to wind direction was eliminated by using an olfactometer that had an air stream approaching the predator from below a gauze screen upon which the predator walked. The steep gradient of odour was obtained by putting a cylinder filled with prey-infested leaves vertically below the screen. Starved predators were arrested in the odour patch by walking more slowly and tortuously than well-fed predators. The latter mites did not show a significant ortho- or klinokinetic response to the presence of odour. Both well-fed and starved predators showed a chemotactic response to steep gradients at the border of the circular odour patch. Predators that happened to walk out of the patch, frequently turned back to it. This response is presumably based on idiothetic information about the predator's immediately previous walking directions, because it occurred in the odour-free zone after passing the steep gradient of prey odour. Right-about turns can help the predator to stay in static odour plumes with steep gradients at the borders. This type of plume is present only close to the odour source. Further away from the source the odour plume tends to move to and fro due to variation in wind direction. For the predator to keep track of these snaking plumes the right-about turns are unlikely to be of any value because the response is of short duration and because the response to a moving plume appeared to be inadequate; by moving the cylinder below the screen (and consequently the odour patch) it was found that the predator turned back even if the odour gradient was made to pass the predator in the same direction as that of the predator's movement.