Precision Grip Control while Walking Down a Stair Step

The aim of this study was to determine whether the internal model regulating grip force (GF)/load force (LF) coordination during a brisk load increase is preserved when the lower extremities produce a perturbation during a single step-down task. We observed the coordination of the vertical ground reaction force (vGRF), GF and LF while holding a handheld object during a single step-down task. The 3 forces (vGRF, GF and LF) decreased during the start of the task. While the subject was descending, LF and GF became dissociated from vGRF and increased in value, probably to anticipate the first foot contact. Coordination of LF and GF was maintained until the maximal vGRF (knee extension). LF peaked in the same time window as vGRF, whereas GF peaked about 70 ms later. This desynchronization, which was previously observed in direct load increase on a handheld object, was interpreted to be a predictive action to ensure the smooth management of the brisk increase in load induced by the lower extremities. Incidentally, in this group, kinematic and dynamic differences were observed between men and women, which may highlight a gender-specific strategy to perform the step-down task. In conclusion, these results suggest that the internal model of precision grip is able to integrate a brisk load change, whatever its origin, and regulate the forces to provide an ideal GF to dampen a brisk load increase and secure the object.     

Rhythmic Manipulation of Objects with Complex Dynamics: Predictability over Chaos

The study of object manipulation has been largely confined to discrete tasks, where accuracy, mechanical effort, or smoothness were examined to explain subjects'' preferred movements. This study investigated a rhythmic manipulation task, which involved continuous interaction with a nonlinear object that led to unpredictable object behavior. Using a simplified virtual version of the task of carrying a cup of coffee, we studied how this unpredictable object behavior affected the selected strategies. The experiment was conducted in a virtual set-up, where subjects moved a cup with a ball inside, modeled by cart-and-pendulum dynamics. Inverse dynamics calculations of the system showed that performing the task with different amplitudes and relative phases required different force profiles and rendered the object''s dynamics with different degrees of predictability (quantified by Mutual Information between the applied force and the cup kinematics and its sensitivity). Subjects (n = 8) oscillated the virtual cup between two targets via a robotic manipulandum, paced by a metronome at 1 Hz for 50 trials, each lasting 45 s. They were free to choose their movement amplitude and relative phase between the ball and cup. Experimental results showed that subjects increased their movement amplitudes, which rendered the interactions with the object more predictable and with lower sensitivity to the execution variables. These solutions were associated with higher average exerted force and lower object smoothness, contradicting common expectations from studies on discrete object manipulation and unrestrained movements. Instead, the findings showed that humans selected strategies with higher predictability of interaction dynamics. This finding expressed that humans seek movement strategies where force and kinematics synchronize to repeatable patterns that may require less sensorimotor information processing.     

Modulation of Force below 1 Hz: Age-Associated Differences and the Effect of Magnified Visual Feedback

Oscillations in force output change in specific frequency bins and have important implications for understanding aging and pathological motor control. Although previous studies have demonstrated that oscillations from 0–1 Hz can be influenced by aging and visuomotor processing, these studies have averaged power within this bandwidth and not examined power in specific frequencies below 1 Hz. The purpose was to determine whether a differential modulation of force below 1 Hz contributes to changes in force control related to manipulation of visual feedback and aging. Ten young adults (25±4 yrs, 5 men) and ten older adults (71±5 yrs, 4 men) were instructed to accurately match a target force at 2% of their maximal isometric force for 35 s with abduction of the index finger. Visual feedback was manipulated by changing the visual angle (0.05°, 0.5°, 1.5°) or removing it after 15 s. Modulation of force below 1 Hz was quantified by examining the absolute and normalized power in seven frequency bins. Removal of visual feedback increased normalized power from 0–0.33 Hz and decreased normalized power from 0.66–1.0 Hz. In contrast, magnification of visual feedback (visual angles of 0.5° and 1.5°) decreased normalized power from 0–0.16 Hz and increased normalized power from 0.66–1.0 Hz. Older adults demonstrated a greater increase in the variability of force with magnification of visual feedback compared with young adults (P = 0.05). Furthermore, older adults exhibited differential force modulation of frequencies below 1 Hz compared with young adults (P<0.05). Specifically, older adults exhibited greater normalized power from 0–0.16 Hz and lesser normalized power from 0.66–0.83 Hz. The changes in force modulation predicted the changes in the variability of force with magnification of visual feedback (R² = 0.80). Our findings indicate that force oscillations below 1 Hz are associated with force control and are modified by aging and visual feedback.     

FTIR and UV spectroscopy in real-time monitoring of S. cerevisiae cell culture

A combination of FTIR and UV spectroscopy is proposed as a novel technique for integrated real-time monitoring of metabolic activity and growth rates of cell cultures, required for systematic studies of cellular low-frequency (LF) electric and magnetic field (EMF) effects. As an example, we investigated simultaneous influence of periodic LF 3D EMFs on a culture of Saccharomyces cerevisiae (baker's yeast) cells. Amplitudes, frequencies and phases of the field components were the variable parameters. Electromagnetic fields were found to efficiently control the activity of the yeast cells, with the resulting CO(2) production rates, as monitored by FTIR spectroscopy, varying by at least one order of magnitude due to the field action. Additionally, population dynamics of the yeast cells was monitored by UV absorption of the yeast culture at λ(prob)?=?320?nm, and compared to the CO(2) production rates. The detected physiologically active frequencies are all below 1?kHz, namely, 800?Hz excitation was effective in reducing the metabolic rates and arresting cell proliferation, whereas 200?Hz excitation was active in accelerating both cell proliferation and overall metabolic rates. The proposed methods produce objective, reliable and quantitative real-time results within minutes and may be used in various tasks that could benefit from a rapid feedback they provide in the form of metabolic and growth rates. Amplitude and frequency dependences of the LF EMF effects from individual field components with different polarizations were recorded and qualitatively interpreted based on a simple model, describing ion diffusion through a membrane channel.     

FTIR and UV spectroscopy in real-time monitoring of S. cerevisiae cell culture

A combination of FTIR and UV spectroscopy is proposed as a novel technique for integrated real-time monitoring of metabolic activity and growth rates of cell cultures, required for systematic studies of cellular low-frequency (LF) electric and magnetic field (EMF) effects. As an example, we investigated simultaneous influence of periodic LF 3D EMFs on a culture of Saccharomyces cerevisiae (baker's yeast) cells. Amplitudes, frequencies and phases of the field components were the variable parameters. Electromagnetic fields were found to efficiently control the activity of the yeast cells, with the resulting CO₂ production rates, as monitored by FTIR spectroscopy, varying by at least one order of magnitude due to the field action. Additionally, population dynamics of the yeast cells was monitored by UV absorption of the yeast culture at λ_prob = 320 nm, and compared to the CO₂ production rates. The detected physiologically active frequencies are all below 1 kHz, namely, 800 Hz excitation was effective in reducing the metabolic rates and arresting cell proliferation, whereas 200 Hz excitation was active in accelerating both cell proliferation and overall metabolic rates. The proposed methods produce objective, reliable and quantitative real-time results within minutes and may be used in various tasks that could benefit from a rapid feedback they provide in the form of metabolic and growth rates. Amplitude and frequency dependences of the LF EMF effects from individual field components with different polarizations were recorded and qualitatively interpreted based on a simple model, describing ion diffusion through a membrane channel.     

Loss of Breathing Modulation of Heart Rate Variability in Patients with Recent and Long Standing Diabetes Mellitus Type II

Healthy subjects under rhythmic breathing have heart interbeat intervals with a respiratory band in the frequency domain that can be an index of vagal activity. Diabetes Mellitus Type II (DM) affects the autonomic nervous system of patients, thus it can be expected changes on the vagal activity. Here, the influence of DM on the breathing modulation of the heart rate is evaluated by analyzing in the frequency domain heart interbeat interval (IBI) records obtained from 30 recently diagnosed, 15 long standing DM patients, and 30 control subjects during standardized clinical tests of controlled breathing at 0.1 Hz, supine rest and standing upright. Fourier spectral analysis of IBI records quantifies heart rate variability in different regions: low-frequencies (LF, 0.04–0.15 Hz), high-frequencies (HF, 0.15–0.4 Hz), and a controlled breathing peak (RP, centered around 0.1 Hz). Two new parameters are introduced: the frequency radius r_f (square root of the sum of LF and HF squared) and β (power of RP divided by the sum of LF and HF). As diabetes evolves, the controlled breathing peak loses power and shifts to smaller frequencies, indicating that heart rate modulation is slower in diabetic patients than in controls. In contrast to the traditional parameters LF, HF and LF/HF, which do not show significant differences between the three populations in neither of the clinical tests, the new parameters r_f and β, distinguish between control and diabetic subjects in the case of controlled breathing. Sympathetic activity that is driven by the baroreceptor reflex associated with the 0.1 Hz breathing modulations is affected in DM patients. Diabetes produces not only a rigid heartbeat with less autonomic induced variability (r_f diminishes), but also alters the coupling between breathing and heart rate (reduced β), due to a progressive decline of vagal and sympathetic activity.     

Increased Force Variability in Chronic Stroke: Contributions of Force Modulation below 1 Hz

Increased force variability constitutes a hallmark of arm disabilities following stroke. Force variability is related to the modulation of force below 1 Hz in healthy young and older adults. However, whether the increased force variability observed post stroke is related to the modulation of force below 1 Hz remains unknown. Thus, the purpose of this study was to compare force modulation below 1 Hz in chronic stroke and age-matched healthy individuals. Both stroke and control individuals (N = 26) performed an isometric grip task to submaximal force levels. Coefficient of variation quantified force variability, and power spectrum density of force quantified force modulation below 1 Hz with a high resolution (0.07 Hz). Analyses indicated that force variability was greater for the stroke group compared with to healthy controls and for the paretic hand compared with the non-paretic hand. Force modulation below 1 Hz differentiated the stroke individuals and healthy controls, as well as the paretic and non-paretic hands. Specifically, stroke individuals (paretic hand) exhibited greater power ∼0.2 Hz (0.07–0.35 Hz) and lesser power ∼0.6 Hz (0.49–0.77 Hz) compared to healthy controls (non-dominant hand). Similarly, the paretic hand exhibited greater power ∼0.2 Hz, and lesser power ∼0.6 Hz than the non-paretic hand. Moreover, variability of force was strongly predicted from the modulation of specific frequencies below 1 Hz (R ² = 0.80). Together, these findings indicate that the modulation of force below 1 Hz provides significant insight into changes in motor control after stroke.     

Digger wasp against crickets. II. An airborne signal produced by a running predator

Summary Females of the digger wasp Liris niger Fabr. hunt crickets to provide food for their offspring by running with high velocity on the ground (>20–50 cm/s). Crickets are able to detect the running wasps by the air particle movement generated by the predator. We measured signals produced by running wasps using a microphone sensitive to air particle velocity. The wasps generated single air puffs with peak air particle velocities of 1–2 cm/s measured close to the running wasp. We measured frequency spectra of the signals containing only components below 50 Hz, with increasing intensities towards lower frequencies, especially below 10 Hz.We measured the air particle movement generated by artificially moved wasps, crickets or a styrofoam dummy of similar size to investigate the effect of velocity and shape of the moving object upon the composition of the signal. The velocity of movement appeared to be important for the intensity and frequency composition of the air particle movement. The shape of the moved body had an influence on the intensity but only little effect on the frequency spectrum. Measurements with a thermistor anemometer showed that a moving object caused air currents lasting longer than 100 ms after passing or approaching the probe. The air particle movements generated by hunting wasps are entirely sufficient with respect to intensity and frequency range to be registered by the filiform hair sensilla upon the cerci of crickets.     

The Role of Left Supplementary Motor Area in Grip Force Scaling

Skilled tool use and object manipulation critically relies on the ability to scale anticipatorily the grip force (GF) in relation to object dynamics. This predictive behaviour entails that the nervous system is able to store, and then select, the appropriate internal representation of common object dynamics, allowing GF to be applied in parallel with the arm motor commands. Although psychophysical studies have provided strong evidence supporting the existence of internal representations of object dynamics, known as “internal models”, their neural correlates are still debated. Because functional neuroimaging studies have repeatedly designated the supplementary motor area (SMA) as a possible candidate involved in internal model implementation, we used repetitive transcranial magnetic stimulation (rTMS) to interfere with the normal functioning of left or right SMA in healthy participants performing a grip-lift task with either hand. TMS applied over the left, but not right, SMA yielded an increase in both GF and GF rate, irrespective of the hand used to perform the task, and only when TMS was delivered 130–180 ms before the fingers contacted the object. We also found that both left and right SMA rTMS led to a decrease in preload phase durations for contralateral hand movements. The present study suggests that left SMA is a crucial node in the network processing the internal representation of object dynamics although further experiments are required to rule out that TMS does not affect the GF gain. The present finding also further substantiates the left hemisphere dominance in scaling GF.     

Experimental Evidence of the Tonic Vibration Reflex during Whole-Body Vibration of the Loaded and Unloaded Leg

Increased muscle activation during whole-body vibration (WBV) is mainly ascribed to a complex spinal and supraspinal neurophysiological mechanism termed the tonic vibration reflex (TVR). However, TVR has not been experimentally demonstrated during low-frequency WBV, therefore this investigation aimed to determine the expression of TVR during WBV.  Whilst seated, eight healthy males were exposed to either vertical WBV applied to the leg via the plantar-surface of the foot, or Achilles tendon vibration (ATV) at 25Hz and 50Hzfor 70s. Ankle plantar-flexion force, tri-axial accelerations at the shank and vibration source, and surface EMG activity of m. soleus (SOL) and m. tibialis anterior (TA) were recorded from the unloaded and passively loaded leg to simulate body mass supported during standing.  Plantar flexion force was similarly augmented by WBV and ATV and increased over time in a load- and frequency dependent fashion. SOL and TA EMG amplitudes increased over time in all conditions independently of vibration mode. 50Hz WBV and ATV resulted in greater muscle activation than 25Hz in SOL when the shank was loaded and in TA when the shank was unloaded despite the greater transmission of vertical acceleration from source to shank with 25Hz and WBV, especially during loading. Low-amplitude WBV of the unloaded and passively loaded leg produced slow tonic muscle contraction and plantar-flexion force increase of similar magnitudes to those induced by Achilles tendon vibration at the same frequencies. This study provides the first experimental evidence supporting the TVR as a plausible mechanism underlying the neuromuscular response to whole-body vibration.     

Modulations of EEG Beta Power during Planning and Execution of Grasping Movements

Although beta oscillations (≈ 13–35 Hz) are often considered as a sensorimotor rhythm, their functional role remains debated. In particular, the modulations of beta power during preparation and execution of complex movements in different contexts were barely investigated. Here, we analysed the beta oscillations recorded with electroencephalography (EEG) in a precued grasping task in which we manipulated two critical parameters: the grip type (precision vs. side grip) and the force (high vs. low force) required to pull an object along a horizontal axis. A cue was presented 3 s before a GO signal and provided full, partial or no information about the two movement parameters. We measured beta power over the centro-parietal areas during movement preparation and execution as well as during object hold. We explored the modulations of power in relation to the amount and type of prior information provided by the cue. We also investigated how beta power was affected by the grip and force parameters.We observed an increase in beta power around the cue onset followed by a decrease during movement preparation and execution. These modulations were followed by a transient power increase during object hold. This pattern of modulations did not differ between the 4 movement types (2 grips ×2 forces). However, the amount and type of prior information provided by the cue had a significant effect on the beta power during the preparatory delay. We discuss how these results fit with current hypotheses on the functional role of beta oscillations.     

A motion analysis marker-based method of determining centre of pressure during two-legged hopping

The fixed position of force plates has led researchers to pursue alternative methods of determining centre of pressure (CoP) location. To date, errors reported using alternative methods to the force plate during dynamic tasks have been high. The aim of this study was to investigate the accuracy of a motion analysis marker-based system to determine CoP during a two-legged hopping task. Five markers were attached to the left and right feet of eight healthy adults (5 females, 3 males, age: 25.0±2.8 years, height: 1.75±0.07 m, mass: 71.3±11.3 kg). Multivariate forward stepwise and forced entry linear regression was used with data from five participants to determine CoP position during quiet standing and hopping at various frequencies. Maximum standard error of the estimate of CoP position was 12 mm in the anteroposterior direction and 8 mm in the mediolateral. Cross-validation was performed using the remaining 3 participants. Maximum root mean square difference between the force plate and marker method was 14 mm for mediolateral CoP and 20 mm for anteroposterior CoP during 1.5 Hz hopping. Differences reduced to a maximum of 7 mm (mediolateral) and 14 mm (anteroposterior) for the other frequencies. The smallest difference in calculated sagittal plane ankle moment and timing of maximum moment was during 3.0 Hz hopping, and largest at 1.5 Hz. Results indicate the marker-based method of determining CoP may be a suitable alternative to a force plate to determine CoP position during a two-legged hopping task at frequencies greater than 1.5 Hz.     

Age impairs sit-to-walk motor performance

Sit-to-walk (STW) is a common functional and transitional task which challenges an individual's postural control systems. As aging is associated with an increased risk of falls during transitional movements, we biomechanically investigated the STW movement task in 12 healthy young and 12 healthy elderly individuals. Performance was evaluated utilizing motion analysis and two force plates. The principal finding of this study was the impaired performance of the healthy older adults. The older adults generated significantly less momentum prior to rising (p=0.011) and further delayed (p<0.001) the initiation of gait until standing more upright (p=0.036). The young adults successfully merged the component tasks shortly after seat-off and displayed significantly greater step length (p<0.001), step velocity (p<0.001), and tolerated greater separation of the center of pressure and center of mass at the end single support phase of the initial step (p=0.001). While the young adults fluidly merged the standing and walking task components, the older adults displayed a conservative movement performance during the STW task thereby limiting threats to their postural stability.     

Cognitive-Motor Interference While Grasping,Lifting and Holding Objects

In daily life, object manipulation is usually performed concurrently to the execution of cognitive tasks. The aim of the present study was to determine which aspects of precision grip require cognitive resources using a motor-cognitive dual-task paradigm. Eighteen healthy participants took part in the experiment, which comprised two conditions. In the first condition, participants performed a motor task without any concomitant cognitive task. They were instructed to grip, lift and hold an apparatus incorporating strain gauges allowing a continuous measurement of the force perpendicular to each contact surface (grip force, GF) as well as the total tangential force applied on the object (load force, LF). In the second condition, participants performed the same motor task while concurrently performing a cognitive task consisting in a complex visual search combined with counting. In the dual-task condition, we found a significant increase in the duration of the preload phase (time between initial contact of the fingers with the apparatus and onset of the load force), as well as a significant increase of the grip force during the holding phase, indicating that the cognitive task interfered with the initial force scaling performed during the preload phase and the fine-tuning of grip force during the hold phase. These findings indicate that these aspects of precision grip require cognitive resources. In contrast, other aspects of the precision grip, such as the temporal coupling between grip and load forces, were not affected by the cognitive task, suggesting that they reflect more automatic processes. Taken together, our results suggest that assessing the dynamic and temporal parameters of precision grip in the context of a concurrent cognitive task may constitute a more ecological and better-suited tool to characterize motor dysfunction in patients.     

Effects of coupled upper limbs movements on postural stabilisation

The preference for in-phase association of coupled cyclic limbs movements is well described (mirror-symmetrical patterns) and this is demonstrated by the ease of performing in-phase movements compared to anti-phase ones. The hypothesis of this study is that the easiest movement patterns are those with minor postural activity. The aim of this study was to describe postural activity in standing subjects in the sagittal and frontal planes during the execution of three upper limbs tasks (single arm, in-phase, anti-phase) at four different frequencies (from 0.6 to 1.2 Hz).We employed six infrared cameras for recording kinematics information, a force platform for measuring forces exerted on the ground, and a system for surface electromyography (SEMG). Outcome measures were: upper limb range of movement and relative-phase, centre of pressure displacement (COP), screw torque (T_z) exerted on the ground, and SEMG recordings of postural muscles (adductor longus, gluteus medius, rectus femoris, and biceps femoris).Our results show that in both the planes the in-phase task resulted in less COP displacement, torque production, and postural muscles involvement than the anti-phase and single arm tasks. This reduced need of postural control could explain the ease of performing in-phase coupled limb movements compared with anti-phase movements.     

Psychophysical principles of discrete event-driven vibrotactile feedback for prostheses

Abstract

Purpose/aim of the study: We aimed to establish psychophysical principles for non-invasive vibrotactile feedback signalling discrete transition events (e.g., extension to flexion) during use of prostheses, especially for the upper limbs.

Materials and methods: Two vibrotactile actuators were used on both upper arms of 10 able-bodied human participants. Absolute thresholds, psychometric functions, and magnitude estimates were measured to equalize the sensation magnitudes for the tested vibrotactile frequencies and skin sites. Then, same-different and pattern recognition tasks were run to evaluate, respectfully, the discrimination and closed-set identification of stimuli with varying parameters (2 frequencies, 2 magnitudes, 2 sites). Finally, parameters of the left/right stimuli were mapped to hypothetical prosthesis events representing object/force and movement type. The stimuli were applied sequentially in accordance with the discrete event-driven feedback paradigm.

Results: Reliable psychophysical models could be established for individual participants as verified by repetitive threshold measurements and relative adjustment of stimulus levels based on sensation magnitudes. Discrimination accuracy was higher for magnitude versus frequency comparisons; and magnitude discrimination accuracy was correlated with magnitude estimate differences. Pattern recognition recall/precision rates decreased from ～0.7 to ～0.5 for sequential delivery of two stimulus patterns to one arm versus to two arms. Using the patterns as two and three consecutive prosthesis events yielded statistically similar performance rates not correlated with magnitude estimate differences.

Conclusions: By careful calibration of stimuli based on psychophysical principles, discrete event-driven vibrotactile feedback can be used to signal manipulated object and movement information with moderate identification rates as shown by confusion matrices.     

Autonomic cardiovascular control in conscious mice

Autonomic cardiovascular control was characterized in conscious, chronically catheterized mice by spectral analysis of arterial pressure (AP) and heart rate (HR) during autonomic blockade or baroreflex modulation of autonomic tone. Both spectra were similar to those obtained in humans, but at approximately 10x higher frequencies. The 1/f relation of the AP spectrum changed to a more shallow slope below 0.1-0.2 Hz. Coherence between AP and HR reached 0.5 or higher below 0.3-0.4 Hz and also above 2.5 Hz. Muscarinic blockade (atropine) or beta-adrenergic blockade (atenolol) did not significantly affect the AP spectrum. Atropine reduced HR variability at all frequencies, but this effect waned above 1 Hz. beta-Adrenergic blockade (atenolol) slightly enhanced the HR variability only above 1 Hz. alpha-Adrenergic blockade (prazosin) reduced AP variability between 0.05 and 3 Hz, most prominently at 0. 15-0.7 Hz. A shift of the autonomic nervous tone by a hypertensive stimulus (phenylephrine) enhanced, whereas a hypotensive stimulus (nitroprusside) depressed AP variability at 1-3 Hz; other frequency ranges of the AP spectrum were not affected except for a reduction below 0.4 Hz after nitroprusside. Variability of HR was enhanced after phenylephrine at all frequencies and reduced after nitroprusside. As with atropine, the reduction with nitroprusside waned above 1 Hz. In conclusion, in mice HR variability is dominated by parasympathetic tone at all frequencies, during both blockade and physiological modulation of autonomic tone. There is a limitation for further reduction but not for augmentation of HR variability from the resting state above 1 Hz. The impact of HR on AP variability in mice is confined to frequencies higher than 1 Hz. Limits between frequency ranges are proposed as 0.15 Hz between VLF (very low frequency range) and LF (low frequency range) and 1.5 Hz between LF and HF (high frequency range).     

Structural studies of the carbohydrate moieties of lectins from potato (Solanum tuberosum) tubers and thorn-apple (Datura stramonium) seeds

1. Methylation analysis of potato (Solanum tuberosum) lectin and thorn-apple (Datura stramonium) lectin confirmed previous conclusions that both glycoproteins contained high proportions of l-arabinofuranosides and lesser amounts of d-galactopyranosides. The arabinofuranosides are present in both lectins as short unbranched chains containing 1→2- and 1→3-linkages, which are known to be linked to hydroxyproline. Galactopyranosides are present as monosaccharides, which are known to be attached to serine, in potato lectin and as both the monosaccharide and the 1→3-linked disaccharide in Datura lectin. 2. Alkaline digestion of potato lectin and subsequent separation of the components by gel filtration led to the isolation of four fractions corresponding to the mono-, di-, tri- and tetra-arabinosides of hydroxyproline. The latter two fractions accounted for over 70% of the total hydroxyproline. 3. Methylation analysis was used to show that the triarabinoside contained only 1→2-linkages between sugars, but that the tetra-arabinoside contained both 1→2- and 1→3-linkages. Direct-insertion mass spectrometry of these compounds using electron impact and chemical ionization, in a comparison with other known structural patterns, was used to determine the sequences of the sugars, which were Araƒ1→2Araƒ1→2Araƒ1→Hyp and Araƒ1→3Araƒ1→2Araƒ1→2Araƒ 1→Hyp. 4. On the basis of optical rotation it had previously been suggested [Allen, Desai, Neuberger & Creeth (1978) Biochem. J. 171, 665–674] that all the arabinose of potato lectin was present as the β-l-furanoside. However, measurement of the optical rotations of the hydroxyprolyl arabinosides showed that whereas the diarabinoside had a molar rotation ([m]) value close to that predicted, the triarabinoside was more dextrorotatory and the tetra-arabinoside was less dextrorotatory than expected. Possible explanations for these findings are that, although the di- and tri-arabinosides contain exclusively β-arabinofuranosides, in the tri-arabinoside, interactions between pentose units lead to an enhanced positive rotation. The tetra-arabinoside, however, is proposed to contain a single α-arabinofuranoside residue, which is responsible for the lower than expected positive rotation. The observed rotation of the tetra-arabinoside was found to be close to the theoretical value predicted on that basis. Furthermore, the action of a specific α-arabinofuranosidase on the tetrasaccharide was to remove a single arabinose residue, presumably the terminal non-reducing sugar, and to produce a product that was indistinguishable on electrophoresis from the triarabinoside. Changes in rotation were compatible with this assumption. 5. It is concluded that the structures of the hydroxyprolyl tri- and tetra-arabinosides of potato lectin are: βAraƒ1→2βAraƒ1→2βAraƒ1→Hyp and αAraƒ1→3βAraƒ1→2βAraƒ 1→2βAraƒ1→Hyp. These are identical with compounds that have been isolated from the insoluble hydroxyproline-rich glycoproteins of plant cell walls.     

Effects of low frequency cyclic mechanical stretching on osteoclastogenesis

Bone cells are continuously exposed to mechanical deformations originating from movement. Mechanical stimulation at fundamental frequencies associated with most frequent normal locomotion (0.167–10 Hz) has been reported to suppress differentiation of osteoclasts. However, the effects of very low frequency (0.01 Hz) stimulation (which could be a frequency component of normal movement and also relevant to locomotion of movement-impaired individuals) on osteoclasts are poorly understood. We examined differentiation of osteoclasts from mouse bone marrow precursors and RAW 264.7 monocytes cultured on an extendable silicone surface that was dynamically stretched at 0.01 Hz. Three stimulation regimes were applied: (i) continuously during 4 days of differentiation, (ii) non-continuously, 8 h/day for 4 days, and (iii) post-differentiation, when stimulation was applied for 24 h after osteoclasts were noted. Low frequency mechanical stimulation did not inhibit osteoclastogenesis. Moreover, the expression of osteoclast marker genes was upregulated in mechanically stimulated cells compared to static control. Conditioned medium collected from osteoclast cultures stimulated non-continuously or post-differentiation induced differentiation of osteoclast precursors plated in standard tissue culture plates. Extracellular signal-regulated kinase (ERK) phosphorylation was increased in mechanically-stimulated cultures compared to static control. Thus, low frequency mechanical stimulation has qualitatively different effects on osteoclast formation compared to stimulation associated with the fundamental frequencies of normal movement.