Acute and Long-Term Effects of Stretching with Whole-Body Vibration on Young's Modulus of the Soleus Muscle Measured Using Shear Wave Elastography.

The effect of whole-body vibration (WBV) stretching on soleus (SOL) muscle stiffness remains unclear. Therefore, we aimed to investigate the acute and long-term effects of stretching with WBV on SOL muscle stiffness. This study employed a repeated-measures experimental design evaluating 20 healthy young males. SOL muscle stretching with WBV was performed for 5 min per day (1 min per set, five sets) over 4 weeks, for 4 days a week. Participants stretched the SOL muscle with ankle dorsiflexion in a loaded flexed knee position on a WBV device. Data were obtained to examine acute effects before stretching, immediately after stretching, and at 5, 10, 15, and 20 min. Moreover, data were obtained to examine the long-term effects before stretching, immediately after the completion of the 4-week stretching program, and at 2 and 4 weeks later. SOL muscle stiffness was measured using Young's modulus with shear wave elastography. The acute effect of SOL muscle stretching with WBV persisted for up to 20 min. Additionally, the long-term effect of stretching was better maintained than the acute effect, which was effective for up to 4 weeks (p < 0.001). Clinically, continuous stretching with WBV may be used to improve SOL muscle stiffness in rehabilitation programs.

Dynamic modulation of enhancer-promoter and promoter-promoter connectivity in gene regulation.

Enhancers are short segments of regulatory DNA that control when and in which cell-type genes should be turned on in response to a variety of extrinsic and intrinsic signals. At the molecular level, enhancers serve as a genomic scaffold that recruits sequence-specific transcription factors and co-activators to facilitate transcription from linked promoters. However, it remains largely unclear how enhancers communicate with appropriate target promoters in the context of higher-order genome topology. In this review, we discuss recent progress in our understanding of the functional interplay between enhancers, genome topology, and the molecular properties of transcription machineries in gene regulation. We suggest that the activities of transcription hubs are highly regulated through the dynamic rearrangement of enhancer-promoter and promoter-promoter connectivity during animal development.

Absolute reliability of Young's modulus of the soleus muscle and Achilles tendon measured using shear wave elastography in healthy young males.

Background: Stiffness of the soleus muscle (SOL) and Achilles tendon (AT) are associated with Achilles tendinitis and medial tibial stress syndrome. Therefore, reliable SOL and AT stiffness measurements are important for monitoring clinical progress. However, little is known about the absolute reliability of the stiffness measurements of SOL and AT in different ankle positions. This study aimed to determine the absolute reliability of the Young's modulus measurements of the SOL and AT in different ankle positions in healthy young males. Methods: This study included 33 healthy young males. SOL and AT stiffnesses were measured using Young's modulus and shear-wave elastography (SWE). Measurements were taken while the participants were kneeling, with their knees flexed to 90°, and the upper body supported by a table. Ultrasound images were recorded at ankle dorsiflexion angles of -10°, 0°, and 10°. The same measurements were repeated 15 min after the first measurement. Bland-Altman plots were used to verify the type or amount of error and 95 % confidence interval of the minimal detectable change (MDC95) values of the measurements. Results: Bland-Altman plots identified that there was no fixed or proportional bias and that there was good agreement between the first- and second-time measurements of the SOL and AT, respectively, among all angles. The MDC95 of the Young's modulus of SOL at -10°, 0°, and 10° of ankle dorsiflexion were 5.6 kPa, 7.0 kPa, and 10.1 kPa, respectively, and AT were 15.8 kPa, 16.4 kPa, and 17.8 kPa, respectively. Conclusion: Young's modulus measurements of the SOL and AT using SWE can be used to quantify elastic properties with high confidence. Clinically, assessing changes in the Young's moduli of the SOL and AT using SWE may help determine the effectiveness of interventions.

Regulatory landscape of enhancer-mediated transcriptional activation.

Enhancers are noncoding regulatory elements that instruct spatial and temporal specificity of gene transcription in response to a variety of intrinsic and extrinsic signals during development. Although it has long been postulated that enhancers physically interact with target promoters through the formation of stable loops, recent studies have changed this static view: sequence-specific transcription factors (TFs) and coactivators are dynamically recruited to enhancers and assemble so-called transcription hubs. Dynamic assembly of transcription hubs appears to serve as a key scaffold to integrate regulatory information encoded by surrounding genome and biophysical properties of transcription machineries. In this review, we outline emerging new models of transcriptional regulation by enhancers and discuss future perspectives.

Enhancer dynamics: Unraveling the mechanism of transcriptional bursting.

Transcriptional bursting is a prevalent feature of gene expression. The transient assembly of transcription factor clusters at regulatory DNAs is critical to control bursting dynamics.

Relationship between the Young's Modulus of the Achilles Tendon and Ankle Dorsiflexion Angle at Maximum Squat Depth in Healthy Young Males.

Background and Objective: Achilles tendon (AT) stiffness can reduce ankle dorsiflexion. However, whether AT stiffness affects the ankle dorsiflexion angle at a maximum squat depth remains unclear. Therefore, we aimed to investigate the relationship between the Young's modulus of the AT and ankle dorsiflexion angle at the maximum squat depth in healthy young males using shear-wave elastography (SWE). Materials and Methods: This cross-sectional study included 31 healthy young males. AT stiffness was measured using the Young's modulus through SWE. The ankle dorsiflexion angle at the maximum squat depth was measured as the angle between the vertical line to the floor and the line connecting the fibula head and the lateral malleolus using a goniometer. Results: Multiple regression analysis identified the Young's modulus of the AT at 10° of ankle dorsiflexion (standardized partial regression coefficient [ß] = -0.461; p = 0.007) and the ankle dorsiflexion angle in the flexed knee (ß = 0.340; p = 0.041) as independent variables for the ankle dorsiflexion angle at maximum squat depth. Conclusions: The Young's modulus of the AT may affect the ankle dorsiflexion angle at the maximum squat depth in healthy young males. Therefore, improving the Young's modulus of the AT may help increase the ankle dorsiflexion angle at maximum squat depth.

Functional coordination between transcription factor clustering and gene activity.

The prevailing view of metazoan gene regulation is that transcription is facilitated through the formation of static activator complexes at distal regulatory regions. Here, we employed quantitative single-cell live-imaging and computational analysis to provide evidence that the dynamic assembly and disassembly process of transcription factor (TF) clusters at enhancers is a major source of transcriptional bursting in developing Drosophila embryos. We further show that the regulatory connectivity between TF clustering and burst induction is highly regulated through intrinsically disordered regions (IDRs). Addition of a poly-glutamine tract to the maternal morphogen Bicoid demonstrated that extended IDR length leads to ectopic TF clustering and burst induction from its endogenous target genes, resulting in defects in body segmentation during embryogenesis. Moreover, we successfully visualized the presence of "shared" TF clusters during the co-activation of two distant genes, which provides a concrete molecular explanation for the newly proposed "topological operon" hypothesis in metazoan gene regulation.

Simulating Knee-Stress Distribution Using a Computed Tomography-Based Finite Element Model: A Case Study.

This study aimed to evaluate the mechanism of progression involved in knee osteoarthritis (OA). We used the computed tomography-based finite element method (CT-FEM) of quantitative X-ray CT imaging to calculate and create a model of the load response phase, wherein the greatest burden is placed on the knee joint while walking. Weight gain was simulated by asking a male individual with a normal gait to carry sandbags on both shoulders. We developed a CT-FEM model that incorporated walking characteristics of individuals. Upon simulating changes owing to a weight gain of approximately 20%, the equivalent stress increased extensively in both medial and lower leg aspects of the femur and increased medio-posteriorly by approximately 230%. As the varus angle increased, stress on the surface of the femoral cartilage did not change significantly. However, the equivalent stress on the surface of the subchondral femur was distributed over a wider area, increasing by approximately 170% in the medio-posterior direction. The range of equivalent stress affecting the lower-leg end of the knee joint widened, and stress on the posterior medial side also increased significantly. It was reconfirmed that weight gain and varus enhancement increase knee-joint stress and cause the progression of OA.

Dynamic interplay between non-coding enhancer transcription and gene activity in development.

Non-coding transcription at the intergenic regulatory regions is a prevalent feature of metazoan genomes, but its biological function remains uncertain. Here, we devise a live-imaging system that permits simultaneous visualization of gene activity along with intergenic non-coding transcription at single-cell resolution in Drosophila. Quantitative image analysis reveals that elongation of RNA polymerase II across the internal core region of enhancers leads to suppression of transcriptional bursting from linked genes. Super-resolution imaging and genome-editing analysis further demonstrate that enhancer transcription antagonizes molecular crowding of transcription factors, thereby interrupting the formation of a transcription hub at the gene locus. We also show that a certain class of developmental enhancers are structurally optimized to co-activate gene transcription together with non-coding transcription effectively. We suggest that enhancer function is flexibly tunable through the modulation of hub formation via surrounding non-coding transcription during development.

The role of clinical engineers in the coronavirus disease 2019 pandemic.

The duties of a clinical engineer (CE) during the coronavirus infection 2019 (COVID-19) pandemic were diverse. The original duties of a CE included operation and maintenance of life support equipment used for respiratory therapy, hemodialysis, and extracorporeal membrane oxygenation. The management of life support equipment is critical. The PB-840 ventilator is equipped with a heat sink system that dissipates internal heat through thermal conduction. Therefore, internal contamination is less likely to occur. The exhalation filter used in the PB- 840 can be used for up to 15 days. It can be used for long periods of time without maintenance, reducing the risk of infection. The PB-840 is a suitable device for patients with COVID-19. Its use in critically ill patients was determined to be a priority. Thus, use of an appropriate device for infection control requires a proper understanding of and familiarity with the device in question.

Mod(mdg4) variants repress telomeric retrotransposon HeT-A by blocking subtelomeric enhancers.

Telomeres in Drosophila are composed of sequential non-LTR retrotransposons HeT-A, TART and TAHRE. Although they are repressed by the PIWI-piRNA pathway or heterochromatin in the germline, the regulation of these retrotransposons in somatic cells is poorly understood. In this study, we demonstrated that specific splice variants of Mod(mdg4) repress HeT-A by blocking subtelomeric enhancers in ovarian somatic cells. Among the variants, we found that the Mod(mdg4)-N variant represses HeT-A expression the most efficiently. Subtelomeric sequences bound by Mod(mdg4)-N block enhancer activity within subtelomeric TAS-R repeats. This enhancer-blocking activity is increased by the tandem association of Mod(mdg4)-N to repetitive subtelomeric sequences. In addition, the association of Mod(mdg4)-N couples with the recruitment of RNA polymerase II to the subtelomeres, which reinforces its enhancer-blocking function. Our findings provide novel insights into how telomeric retrotransposons are regulated by the specific variants of insulator proteins associated with subtelomeric sequences.

Molecular architecture of enhancer-promoter interaction.

Temporal and spatial specificity of gene expression is highly regulated through a rich milieu of regulatory DNAs embedded in the genome. Enhancers represent a major class of regulatory DNAs that consist of a cluster of binding sites for sequence-specific transcription factors and are thought to facilitate recruitment of transcription machinery to their target promoters from remote locations. It has been over four decades since the discovery of prototypic simian virus 40 enhancer, yet the nature of enhancer-promoter interaction still remains an outstanding mystery in gene regulation. The aim of this review is to comprehensively overview molecular mechanisms underlying enhancer-promoter interaction including the roles of looping factors, higher-order genome topology, and dynamic clustering of transcription apparatus within a nucleus. We propose that cooperative interplay between 'looping' and 'hub' permits distal enhancers to specifically and dynamically modulate target gene expression over large distances.

Dynamic modulation of enhancer responsiveness by core promoter elements in living Drosophila embryos.

Regulatory interactions between enhancers and core promoters are fundamental for the temporal and spatial specificity of gene expression in development. The central role of core promoters is to initiate productive transcription in response to enhancer's activation cues. However, it has not been systematically assessed how individual core promoter elements affect the induction of transcriptional bursting by enhancers. Here, we provide evidence that each core promoter element differentially modulates functional parameters of transcriptional bursting in developing Drosophila embryos. Quantitative live imaging analysis revealed that the timing and the continuity of burst induction are common regulatory steps on which core promoter elements impact. We further show that the upstream TATA also affects the burst amplitude. On the other hand, Inr, MTE and DPE mainly contribute to the regulation of the burst frequency. Genome editing analysis of the pair-rule gene fushi tarazu revealed that the endogenous TATA and DPE are both essential for its correct expression and function during the establishment of body segments in early embryos. We suggest that core promoter elements serve as a key regulatory module in converting enhancer activity into transcription dynamics during animal development.

Sway and Acceleration Changes of the Center of Mass during Walking Stance Phase before and after Total Knee Arthroplasty.

Elucidating the sway and changes in the acceleration of center of mass (COM) during walking is important for effective gait training and rehabilitation. The objective of this study was to verify the improvement in gait before and after total knee arthroplasty (TKA) from COM sway and the changes in the acceleration of COM during the stance phase of walking. This study included 13 patients (1 male and 12 females) with medial knee osteoarthritis who were hospitalized for TKA. The COM sway during the stance phase of walking was evaluated using root mean square (RMS) normalized by walking speed, and the changes in acceleration were further verified. Lateral and vertical RMS showed significant differences between preoperative and postoperative states and demonstrated low values after TKA. The lateral acceleration at the latter part of the early stance phase demonstrated a significant difference between preoperative and postoperative states. A significant difference was also observed in the lateral acceleration in the late stance phase between the two groups. Improvement in pain and alignment after TKA reduced the lateral sway of COM and the changes in acceleration during the gait stance phase, which is speculated to lead to improvement in gait and prevention of falls.

Influence of the Amount of Change in Quadriceps Tendon Young's Modulus on Amount of Change in Walking Speed before and after Total Knee Arthroplasty.

Background and Objectives: Walking speed after total knee arthroplasty (TKA) is an important outcome. However, the effect of quadriceps tendon stiffness on walking speed remains unclear. This study aimed to clarify the influence of the amount of change in quadriceps tendon stiffness on the degree of change in walking speed before and after TKA. Materials and Methods: Sixteen patients who underwent TKA for knee osteoarthritis participated in this study (median age: 74.0 years (interquartile range: 64.5-75.8)). Shear-wave elastography was deployed to measure quadriceps tendon stiffness using Young's modulus. A motion analysis system was used to assess kinematic parameters and walking speed. Participants' knee circumference, range of motion, extension strength, one-leg standing time, walking pain level, and activity level were measured preoperatively and one year after TKA, and changes in values were calculated. We used path analysis to clarify the influence of the amount of change in the quadriceps tendon Young's modulus on the change in walking speed. Results: The quadriceps tendon Young's modulus negatively affected the knee flexion angle during swing (standardized partial regression coefficients (ß) = -0.513, p = 0.042). The knee flexion angle during swing positively affected step length (ß = 0.586, p = 0.017). Step length positively affected cadence (ß = 0.733, p = 0.001). Step length and cadence positively affected walking speed (ß = 0.563, p < 0.001, ß = 0.502, p < 0.001, respectively). Conclusions: The amount of change in the quadriceps tendon Young's modulus may affect the degree of change in walking speed after TKA through the amount of change in the knee flexion angle during swing, step length, and cadence. Clinically, reducing quadriceps tendon stiffness can be addressed in rehabilitation programs to increase walking speed after TKA.

Effect of Training With the Hybrid Assistive Limb on Gait Cycle Kinematics After Total Knee Arthroplasty.

INTRODUCTION: Reportedly, wearable robots, such as the hybrid assistive limb (HAL), are effective in the functional recovery of various locomotor disabilities, including disrupted walking, restricted range of motion, and muscle weakness. However, the effect of walking exercises with a HAL on the kinematic and kinetic variables of lower limb joint function is not yet fully understood. Therefore, the purpose of this study was to elucidate the effect of HAL on the kinematic and kinetic variables of lower limb function in patients 5 weeks after total knee arthroplasty (TKA). MATERIALS AND METHODS: Nine patients (ten knees) in the HAL training group and nine patients (nine knees) in the control group underwent TKA. HAL training was initiated 1-5 weeks after TKA, and general rehabilitation was performed in the control group. Gait analysis was performed on each patient using a motion analysis system at 5 weeks after TKA. We compared the effects of the joint angles of the walking cycle between groups, and investigated the effect of the walking cycle's joint angles on step length. RESULTS: In the HAL group, the odds ratio of hip extension was as large as 1.741, while that of knee swing was as large as 1.501. These 2 variables were significant between the 2 groups. Knee swing and varus significantly affected step length. CONCLUSIONS: Our results suggest that training by wearing HAL after TKA increased the mobility of the knee and hip joints during early postoperative walking, and that walking ability was improved by increasing the step length.

Interpretation of causal relationship between quadriceps tendon Young's modulus and gait speed by structural equation modeling in patients with severe knee osteoarthritis.

PURPOSE: To clarify the causal relationship between quadriceps tendon stiffness and gait speed in patients with severe knee osteoarthritis (OA) using structural equation modeling. METHODS: Participants were 36 patients with knee OA (median age, 75.0 [interquartile range, 67.3-76.0] years; Kellgren-Lawrence grade 3 or 4). We measured quadriceps tendon stiffness using Young's modulus by ShearWave Elastography. Gait speed and kinematics parameters were measured using a motion analysis system. Additional data collected for each patient included age, sex, height, body weight, body mass index, femorotibial angle, knee range of motion, knee extension torque, and pain. We performed structural equation modeling for interpretation of the causal relationship. RESULTS: The comparative fit index of the structural equation modeling was 0.990. Quadriceps tendon Young's modulus was a predictor of maximum knee flexion angle during the swing phase (standardized partial regression coefficients [ß] = -0.67, P < 0.001). Maximum knee flexion angle during the swing phase was a predictor of cadence and step length (ß values 0.35 and 0.55, P = 0.035 and <0.001, respectively). Cadence and step length were predictors of gait speed (ß values 0.50 and 0.63, P < 0.001 and <0.001, respectively). CONCLUSION: Our results showed a causal relationship between quadriceps tendon stiffness and gait speed in patients with severe knee OA. Quadriceps tendon Young's modulus can affect gait speed through the maximum knee flexion angle during the swing phase, cadence, and step length. Adding therapeutic intervention to decrease the quadriceps tendon Young's modulus may lead to increased gait speed.

A Simulation Case Study of Knee Joint Compressive Stress during the Stance Phase in Severe Knee Osteoarthritis Using Finite Element Method.

Background and Objectives: Medial knee osteoarthritis is known to increase the mechanical load on the medial compartment of the knee joint during walking; however, it is not visually understood how much the mechanical load increases nor where in the medial compartment of the knee joint that load is focused. Therefore, we conducted a simulation study to determine the location and amount of the mechanical load in the medial compartment of the knee joint during the stance phase. Materials and Methods: Subject was a patient with right medial knee osteoarthritis. Computed tomography imaging and gait analysis were performed on subject. The CT image of the right knee was calculated using finite element analysis software. Since this software can set the flexion angle arbitrarily while maintaining the nonuniform material properties of the bone region, the model is constructed by matching the knee joint extension image obtained by CT to the loading response phase of gait analysis. The data of muscle exertion tension and vertical ground reaction force were inserted into the knee joint model created from the computed tomography-based finite element method, and the knee joint compressive stress was calculated. Results: With regard to compressive stress, the tibia showed high stress at 4.10 to 5.36 N/mm2. The femur showed high stress at 4.00 to 6.48 N/mm2. The joint compressive stress on the medial compartment of the knee joint was found to concentrate on the edge of the medial tibial condyle in the medial knee osteoarthritis subject. Conclusions: The measurement method of knee joint compressive stress by computed tomography-based finite element method can visually be a reliable method of measuring joint compressive stress in the medial knee osteoarthritis. This reflects the clinical findings because concentration of stress on the medial knee joint was observed at the medial osteophyte.

Dynamic regulation of anterior-posterior patterning genes in living Drosophila embryos.

Expression of the gap and pair-rule genes plays an essential role in body segmentation during Drosophila embryogenesis.1-5 However, it remains unclear how precise expression patterns of these key developmental genes arise from stochastic transcriptional activation at the single-cell level. Here, I employed genome-editing and live-imaging approaches to comprehensively visualize regulation of the gap and pair-rule genes at the endogenous loci. Quantitative image analysis revealed that the total duration of active transcription (transcription period) is a major determinant of spatial patterning of gene expression in early embryos. The length of the transcription period is determined by the continuity of bursting activities in individual nuclei, with the core expression domain producing more bursts than boundary regions. Each gene exhibits a distinct rate of nascent RNA production during transcriptional bursting, which contributes to gene-to-gene variability in the total output. I also provide evidence for "enhancer interference," wherein a distal weak enhancer interferes with transcriptional activation by a strong proximal enhancer to downregulate the length of the transcription period without changing the transcription rate. Analysis of the endogenous hunchback (hb) locus revealed that the removal of the distal shadow enhancer induces strong ectopic transcriptional activation, which suppresses refinement of the initial broad expression domain into narrower stripe patterns at the anterior part of embryos. This study provides key insights into the link between transcriptional bursting, enhancer-promoter interaction, and spatiotemporal patterning of gene expression during animal development.

SARS-CoV-2 Leakage From the Gas Outlet Port During Extracorporeal Membrane Oxygenation for COVID-19.

Patients with the coronavirus disease 2019 (COVID-19) sometimes develop refractory respiratory failure and may require venovenous extracorporeal membrane oxygenation (VV-ECMO). It is known that the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is sometimes present in the blood of COVID-19 patients. VV-ECMO is often used for several weeks, and plasma leaks can occur, albeit rarely. Hence, in terms of infection control, a concern is that SARS-CoV-2 may leak from the gas outlet port of the oxygenator during ECMO support of critically ill COVID-19 patients. The aim of this study was to clarify whether SARS-CoV-2 leaks from the oxygenator during ECMO support. Five patients with critical COVID-19 pneumonia were placed on VV-ECMO. Silicone-coated polypropylene membrane oxygenators were used in the ECMO circuits for these patients. SARS-CoV-2 ribonucleic acid (RNA) was measured by quantitative reverse transcription polymerase chain reaction in serum and at the gas outlet port of the ECMO circuit at the time of circuit replacement or liberation from ECMO. SARS-CoV-2 RNA was detected in the gas outlet port of the ECMO circuit for three of the five patients. None of the medical staff involved in the care of these five patients has been infected with COVID-19. In conclusion, SARS-CoV-2 could leak to the gas outlet port of the ECMO circuit through silicone-coated polypropylene membranes during ECMO support of critically ill COVID-19 patients.