Tissue polarity points from cells that have higher Frizzled levels towards cells that have lower Frizzled levels

Background: The frizzled (fz) gene of Drosophila encodes the founding member of the large family of receptors for the Wnt family of signaling molecules. It was originally studied in the adult epidermis, where it plays a key role in the generation of tissue polarity. Mutations in components of the fz signal transduction pathway disrupt tissue polarity; on the wing, hairs normally point distally but their polarity is altered by these mutations.Results: We devised a method to induce a gradient of fz expression with the highest levels near the distal wing tip. The result was a large area of proximally pointing hairs in this region. This reversal of polarity was seen when fz expression was induced just before the start of hair morphogenesis when polarity is established, suggesting that the gradient of Fz protein acted fairly directly to reverse hair polarity. A similar induction of the dishevelled (dsh) gene, which acts cell autonomously and functions downstream of fz in the generation of tissue polarity, resulted in a distinct tissue polarity phenotype, but no reversal of polarity; this argues that fz signaling was required for polarity reversal. Furthermore, the finding that functional dsh was required for the reversal of polarity argues that the reversal requires normal fz signal transduction.Conclusions: The data suggest that cells sense the level of Fz protein on neighboring cells and use this information in order to polarize themselves. A polarizing signal is transmitted from cells with higher Fz levels to cells with lower levels. Our observations enable us to propose a general mechanism to explain how Wnts polarize target cells.     

Acetylation increases access of remodelling complexes to their nucleosome targets to enhance initiation of V(D)J recombination

Targeted chromatin remodelling is essential for many nuclear processes, including the regulation of V(D)J recombination. ATP-dependent nucleosome remodelling complexes are important players in this process whose activity must be tightly regulated. We show here that histone acetylation regulates nucleosome remodelling complex activity to boost RAG cutting during the initiation of V(D)J recombination. RAG cutting requires nucleosome mobilization from recombination signal sequences. Histone acetylation does not stimulate nucleosome mobilization per se by CHRAC, ACF or their catalytic subunit, ISWI. Instead, we find the more open structure of acetylated chromatin regulates the ability of nucleosome remodelling complexes to access their nucleosome templates. We also find that bromodomain/acetylated histone tail interactions can contribute to this targeting at limited concentrations of remodelling complex. We therefore propose that the changes in higher order chromatin structure associated with histone acetylation contribute to the correct targeting of nucleosome remodelling complexes and this is a novel way in which histone acetylation can modulate remodelling complex activity.     

The effect of plantar flexor muscle fatigue on postural control

Objective

Previous studies have demonstrated that ankle muscle fatigue alters postural sway. Our aim was to better understand postural control mechanisms during upright stance following plantar flexor fatigue.

Method

Ten healthy young volunteers, 25.7 ± 2.2 years old, were recruited. Foot center-of-pressure (CoP) displacement data were collected during narrow base upright stance and eyes closed (i.e. blindfolded) conditions. Subjects were instructed to stand upright and as still as possible on a force platform under five test conditions: (1) non-fatigue standing on firm surface; (2) non-fatigue standing on foam; (3) ankle plantar flexor fatigue, standing on firm surface; (4) ankle plantar flexor fatigue, standing on foam; and (5) upper limb fatigue, standing on firm surface. An average of the ten 30-s trials in each of five test conditions was calculated to assess the mean differences between the trials. Traditional measures of postural stability and stabilogram-diffusion analysis (SDA) parameters were analyzed.

Results

Traditional center of pressure parameters were affected by plantar flexor fatigue, especially in the AP direction. For the SDA parameters, plantar flexor fatigue caused significantly higher short-term diffusion coefficients, and critical displacement in both mediolateral (ML) and anteroposterior (AP) directions. Long-term postural sway was different only in the AP direction.

Conclusions

Localized plantar flexor fatigue caused impairment to postural control mainly in the Sagittal plane. The findings indicate that postural corrections, on average, occurred at a higher threshold of sway during plantar flexor fatigue compared to non-fatigue conditions.     

Effect of muscle fatigue on the compliance of the gastrocnemius medialis tendon and aponeurosis

The aim of the present study was to examine whether or not the compliance of the gastrocnemius medialis (GM) tendon and aponeurosis is influenced by submaximal fatiguing efforts. Fourteen elderly male subjects performed isometric maximal voluntary plantarflexion contractions (MVC) on a dynamometer before and after two fatiguing protocols. The protocols consisted of: (1) submaximal concentric isokinetic contractions (70% isokinetic MVC) at 60 degrees /s and (2) a sustained isometric contraction (40% isometric MVC) until failure to hold the defined moment. Ultrasonography was used to determine the elongation and strain of the GM tendon and aponeurosis. To account for the axis misalignment between ankle and dynamometer, the kinematics of the leg were captured at 120 Hz. The maximum moment decreased from 85.9+/-17.9 Nm prior fatigue to 79.2+/-19 Nm after isokinetic fatigue and to 69.9+/-16.4 Nm after isometric fatigue. The maximal strain of the GM tendon and aponeurosis before fatigue, after isokinetic and after isometric fatigue were 4.9+/-1.1%, 4.4+/-1.1% and 4.3+/-1.1% respectively. Neither the strain nor the elongation showed significant differences before and after each fatiguing task at any 100 N step of the calculated tendon force. This implies that the compliance was not altered after either the isokinetic or the isometric fatiguing task. Therefore it was concluded that the strains during the performed submaximal fatiguing tasks, were too small to provoke any structural changes in tendon and aponeurosis.     

Ultrasound does not provide reliable results for the measurement of the patellar tendon cross sectional area

PurposeThis study examined the reliability of patellar tendon cross sectional area (CSA) measurement using brightness mode ultrasonography.MethodsThe patellar tendon CSA of fourteen participants was examined on two different days and at three different positions (proximal, median and distal). Five trials per day were conducted in each position, replacing the ultrasound probe on every trial. The images were examined by three different and equally experienced observers. We compared the mean of the five trials in each position to examine the day, observer and position effect. Further, Bland and Altman plots, root mean square (RMS) differences and intraclass correlation coefficients (ICC) were calculated.ResultsThere was a significant (p < 0.05) day, observer and position effect on the CSA, while the average ICC was 0.592. The Bland and Altman plots showed that differences between conditions or groups, should be in average lower than 37% or higher than 55% of the patellar tendon CSA to be important for clinical or intervention studies.ConclusionOur findings show low reliability of the method, which resulted from the low clarity and unclear visibility of tissue boundaries in the ultrasound images. Therefore, the measurement of the CSA of the patellar tendon using ultrasound does not provide accurate and reliable results.     

Time-related alterations of superoxide radical levels in diverse organs of bile duct-ligated rats

The time-related alterations of superoxide radical measured in vivo by employing an ultrasensitive fluorescent assay in the liver, intestine, kidney and brain of rats with experimentally induced obstructive jaundice was investigated. Eighteen rats were randomly divided into Group A, rats subjected to sham operation, and Group B, rats subjected to bile duct ligation (BDL). Three rats from each group were subsequently killed at different time points post-operatively (1, 5 and 10 days). As compared to sham-operated, BDL rats showed a gradual increase with time of superoxide radical in the intestine, liver, kidney and brain: for animals sacrificed on the 1^st, 5^th and 10^th day the increase was 45%, 50% and 96% in the liver, 76%, 81% and 118% in the intestine, 64%, 71% and 110% in the kidney and 76%, 95% and 142% in the brain, respectively. This study provides direct evidence of an early appearance of oxidative stress in diverse organs, implying a uniform systemic response to biliary obstruction and emphasizing the need of early bile flow restoration.     

Influence of pole plant time on the performance of a special jump and plant exercise in the pole vault

The purpose of this study was to examine the effect of the timing of the pole plant during the stance phase of the jump on the energy level of the vaulter/pole system at take-off for a special pole vault take-off exercise (Jagodin). We hypothesised that an earlier pole plant would increase the pole energy at take-off compared to the energy decrease of the vaulter during the jump and plant complex and so lead to a higher total energy of the vaulter/pole system at take-off. Six male pole vaulters experienced three Jagodins each with different pole plant time building three groups of vaults (early, intermediate, late pole plant). Kinematic data of vaulter and pole were recorded, as were ground reaction forces measured at the end of the pole under the planting box and under the take-off foot. These measurements allowed the energy exchange between the vaulter and pole to be determined. We found neither statistical significant differences in the mechanical energy level of the vaulter/pole system during take-off between the three groups nor a relationship between the timing of the pole plant and the energy level of the vaulter-pole system during take-off. We conclude that although the timing of the pole plant influences the interactions between the vaulter, the pole, and the ground, it does not affect the athlete's performance. Although a late pole plant decreases the loss of energy by the vaulter during the take-off, this is counterbalanced by a decrease in the energy stored in the pole at take-off.     

Relationship of p21‐activated kinase (PAK) and filopodia to persistence and oncogenic transformation

Previously, we found that oncogenically transformed cells had fewer filopodia and more large, p21‐activated kinase (PAK)‐dependent features than normal cells. These large protrusions (LPs) were increased in cells expressing RhoA^N19 with Cdc42‐associated kinase (ACK). Here, we determine how GTPase‐mediated mechanisms of focal contact (FC) regulation affect these protrusions. Constructs encoding various proteins were introduced into cells which were then studied by microscopy and computerized image processing and analysis. Constructs that prevented PAK recruitment by PAK‐interacting exchange factor (PIX) or restricted PAK residence time on FCs decreased both protrusions. Thus, filopodia were also PAK‐dependent. A comparison of FC distribution in cells expressing PAK in the presence or absence of PAK kinase inhibitor domain (KID) suggested that PAK enlarged FCs without affecting the prevalence of either protrusion. KID or Nck expression increased LPs but not filopodia. Nck failed to synergize with KID or ACK and RhoA^N19 in enhancing LPs. Nck and KID synergistically enhanced filopodia, possibly because Nck recruited PAK to FCs while KID prevented their dissociation by PAK‐mediated autophosphorylation. Coexpression of Nck, ACK, and RhoA^N19 abrogated filopodia and replicated the transformed phenotype. Since Nck recruitment of PAK is implicated in persistence of directional movement, we studied the PAK–Nck interface. Filopodia were eliminated by the Nck PAK‐binding domain and LPs by the PAK Nck‐binding domain. The results suggested that filopodia formation has more stringent requirements than LP formation, and Nck and PAK are used differently in the protrusions. Loss of filopodia in transformed cells may reflect defective regulation of GTPase mechanisms. J. Cell. Physiol. 220: 576–585, 2009. © 2009 Wiley‐Liss, Inc.     

Plasticity of human Achilles tendon mechanical and morphological properties in response to cyclic strain

The purpose of the current study in combination with our previous published data (Arampatzis et al., 2007) was to examine the effects of a controlled modulation of strain magnitude and strain frequency applied to the Achilles tendon on the plasticity of tendon mechanical and morphological properties. Eleven male adults (23.9±2.2 yr) participated in the study. The participants exercised one leg at low magnitude tendon strain (2.97±0.47%), and the other leg at high tendon strain magnitude (4.72±1.08%) of similar frequency (0.5 Hz, 1 s loading, 1 s relaxation) and exercise volume (integral of the plantar flexion moment over time) for 14 weeks, 4 days per week, 5 sets per session. The exercise volume was similar to the intervention of our earlier study (0.17 Hz frequency; 3 s loading, 3 s relaxation) allowing a direct comparison of the results. Before and after the intervention ankle joint moment has been measured by a dynamometer, tendon–aponeurosis elongation by ultrasound and cross-sectional area of the Achilles tendon by magnet resonance images (MRI). We found a decrease in strain at a given tendon force, an increase in tendon–aponeurosis stiffness and tendon elastic modulus of the Achilles tendon only in the leg exercised at high strain magnitude. The cross-sectional area (CSA) of the Achilles tendon did not show any statistically significant (P>0.05) differences to the pre-exercise values in both legs. The results indicate a superior improvement in tendon properties (stiffness, elastic modulus and CSA) at the low frequency (0.17 Hz) compared to the high strain frequency (0.5 Hz) protocol. These findings provide evidence that the strain magnitude applied to the Achilles tendon should exceed the value, which occurs during habitual activities to trigger adaptational effects and that higher tendon strain duration per contraction leads to superior tendon adaptational responses.     

1998 Basmajian Student Award Paper: Movement patterns after anterior cruciate ligament injury: a comparison of patients who compensate well for the injury and those who require operative stabilization

The purpose of this study was to describe kinematic and kinetic differences between a group of ACL deficient subjects who were grouped according to functional ability. Sixteen patients with complete ACL rupture were studied; eight subjects had instability with activities of daily living (non-copers) and eight subjects had returned to all pre-injury activity without limitation (copers). Three-dimensional joint kinematics and kinetics were collected from the knee and ankle during walking, jogging and going up and over a step. Results showed that both groups mitigated the force with which they contacted the floor but non-copers consistently demonstrated less knee flexion in the involved limb. The copers used joint kinematics similar to those of their uninvolved knees and similar to knee motions reported in uninjured subjects. The reduced knee motion in the involved knee of the non-copers did not correlate directly with quadriceps femoris muscle weakness.

The data suggest that the non-copers utilize a stabilization strategy which stiffens the knee joint which not only is unsuccessful but may lead to excessive joint contact forces which have the potential to damage articular structures. The copers use a strategy which permits normal knee kinematics and bodes well for joint integrity.