Combined Effects of Static and Dynamic Stretching on the Muscle-Tendon Unit Stiffness and Strength of the Hamstrings.

ABSTRACT: Takeuchi, K, Nakamura, M, Matsuo, S, Samukawa, M, Yamaguchi, T, and Mizuno, T. Combined effects of static and dynamic stretching on the muscle-tendon unit stiffness and strength of the hamstrings. J Strength Cond Res 38(4): 681-686, 2024-Combined static and dynamic stretching for 30 seconds is frequently used as a part of a warm-up program. However, a stretching method that can both decrease muscle-tendon unit (MTU) stiffness and increase muscle strength has not been developed. The purpose of this study was to examine the combined effects of 30 seconds of static stretching at different intensities (normal-intensity static stretching [NS] and high-intensity static [HS]) and dynamic stretching at different speeds (low-speed dynamic [LD] and high-speed dynamic stretching [HD]) on the MTU stiffness and muscle strength of the hamstrings. Thirteen healthy subjects (9 men and 4 women, 20.9 ± 0.8 years, 169.3 ± 7.2 cm, 61.1 ± 8.2 kg) performed 4 types of interventions (HS-HD, HS-LD, NS-HD, and NS-LD). Range of motion (ROM), passive torque, MTU stiffness, and muscle strength were measured before and immediately after interventions by using an isokinetic dynamometer machine. In all interventions, the ROM and passive torque significantly increased (p < 0.01). Muscle-tendon unit stiffness significantly decreased in HS-HD and HS-LD (both p < 0.01), but there was no significant change in NS-HD (p = 0.30) or NS-LD (p = 0.42). Muscle strength significantly increased after HS-HD (p = 0.02) and NS-LD (p = 0.03), but there was no significant change in HS-LD (p = 0.23) or NS-LD (p = 0.26). The results indicated that using a combination of 30 seconds of high-intensity static stretching and high-speed dynamic stretching can be beneficial for the MTU stiffness and muscle strength of the hamstrings.

Acute and Prolonged Effects of 300 sec of Static, Dynamic, and Combined Stretching on Flexibility and Muscle Force.

Static stretching (SS), dynamic stretching (DS), and combined stretching (CS; i.e., DS+SS) are commonly performed as warm-up exercises. However, the stretching method with the greatest effect on flexibility and performance remains unclear. This randomized crossover trial examined acute and prolonged effects of SS, DS, and CS on range of motion (ROM), peak passive torque (PPT), passive stiffness, and isometric and concentric muscle forces. Twenty healthy young men performed 300 sec of active SS, DS, or CS (150-sec SS followed by 150-sec DS and 150-sec DS followed by 150-sec SS) of the right knee flexors on four separate days, in random order. Subsequently, we measured ROM, PPT, and passive stiffness during passive knee extension. We also measured maximum voluntary isometric and concentric knee flexion forces and surface electromyographic activities during force measurements immediately before, immediately after, and 20 and 60 min after stretching. All stretching methods significantly increased ROM and PPT, while significantly decreasing isometric knee flexion force (all p < 0.05). These changes lasted 60 min after all stretching methods; the increases in ROM and PPT and the decreases in isometric muscle force were similar. All stretching methods also significantly decreased passive stiffness immediately after stretching (all p < 0.05). Decreases in passive stiffness tended to be longer after CS than after SS or DS. Concentric muscle force was decreased after SS and CS (all p < 0.05). On the other hand, concentric muscle force was unchanged after DS, while the decreases in surface electromyographic activities during concentric force measurements after all stretching methods were similar. Our results suggest that 300 sec of SS, DS, and CS have different acute and prolonged effects on flexibility and muscle force.

Effects of Speed and Amplitude of Dynamic Stretching on the Flexibility and Strength of the Hamstrings.

Dynamic stretching for more than 90 seconds is useful for improving muscle strength, although dynamic stretching for 30 seconds or less is commonly used in sports settings. The effects of dynamic stretching are influenced by the speed and amplitude of stretching, but no study examined these factors for 30 seconds of dynamic stretching. Therefore, the purpose of the present study was to examine the effects of speed (fast- or slow-speed) and amplitude (normal- or wide amplitude) of dynamic stretching for 30 seconds on the strength (peak torque during maximum isokinetic concentric contraction) and flexibility (range of motion, passive torque at maximum knee extension angle, and muscle-tendon unit stiffness) of the hamstrings. The passive torque and muscle-tendon unit stiffness reflect stretching tolerance and viscoelastic properties of the hamstrings, respectively. Fifteen healthy participants performed 4 types of 30 seconds of dynamic stretching. The muscle strength and flexibility were measured before and immediately after the dynamic stretching. The range of motion did not change after dynamic stretching at low speed and normal amplitude (p = 0.12, d = 0.59, 103.3%), but it was increased by other interventions (p < 0.01, d = 0.90-1.25, 104.5-110.1%). In all interventions, the passive torque increased (main effect for time, p < 0.01, d = 0.51 - 0.74, 111.0 - 126.9%), and muscle-tendon unit stiffness did not change. The muscle strength increased only after dynamic stretching at fast speed with normal amplitude (p < 0.01, d = 0.79, 107.1%). The results of the present study indicated that 30 seconds of dynamic stretching at fast speed and with normal amplitude can be beneficial for the measured parameters.

Effects of High-Intensity Stretch with Moderate Pain and Maximal Intensity Stretch without Pain on Flexibility.

In this study, we aimed to identify the time course effects of different intensities of static stretch (SST) (maximal intensity without pain vs. high-intensity with moderate pain) on flexibility. This study included 16 healthy students (8 men and 8 women) who performed 1) 5-minute SST at 100%, 2) 110%, and 3) 120% intensity, as well as 4) no stretching (control) in a random sequence on four separate days. Static passive torque (SPT), hamstring electromyography (EMG), and pain intensity were continuously recorded during SST. We assessed markers of stiffness, range of motion (ROM), and maximal dynamic passive torque (DPTmax) before SST and 0, 15, 30, 45, 60, 75, and 90 minutes after SST. Stiffness decreased and ROM and DPTmax increased significantly immediately after SST at the three different intensity levels (p < 0.05). The effects of SST at 120% intensity were stronger and lasted longer than the effects of SST at 110% and 100% intensity (stiffness: -17%, -9%, and -7%, respectively; ROM: 14%, 10%, and 6%, respectively; DPTmax: 15%, 15%, and 9%, respectively). SPT decreased after SST at all intensities (p < 0.05). SST at 120% intensity caused a significantly greater reduction in SPT than SST at 100% intensity (p < 0.05). Pain intensity and EMG activity increased immediately after the onset of SST at 120% intensity (p < 0.05), although these responses were attenuated over time. Stretching intensity significantly correlated with the degree of change in ROM and stiffness (p < 0.05). These results support our hypothesis that stretch-induced flexibility is amplified and prolonged with an increase in stretch intensity beyond the pain threshold. Additional studies with more participants and different demographics are necessary to examine the generalizability of these findings.

Acute and chronic effects of static stretching at 100% versus 120% intensity on flexibility.

PURPOSE: The acute effects of static stretching have been frequently studied, but the chronic effects have not been studied concurrently. Thus, this study aimed to investigate both the acute and chronic effects of static stretching at different intensities on flexibility. METHODS: Twenty-three healthy men were randomly assigned to perform 1 min of static stretching 3 days/week for 4 weeks at 100% intensity (n = 12) or 120% intensity (n = 11). The acute effects of stretching were assessed by measuring the range of motion (ROM), peak passive torque, and passive stiffness before and after every stretching session; the chronic effects of stretching were assessed by measuring these outcomes at baseline and after 2 and 4 weeks of stretching. RESULTS: Compared with the 100% intensity group, the 120% intensity group had significantly greater acute increases in ROM after all 12 sessions, a significantly greater decrease in passive stiffness after 11 of 12 sessions, and a significantly greater increase in peak passive torque after six of 12 sessions. Regarding the chronic effects, ROM was significantly increased in both groups after 2 and 4 weeks of stretching. Peak passive torque significantly increased in the 100% intensity group after 2 and 4 weeks of stretching, and after 4 weeks in the 120% intensity group. CONCLUSION: Stretching at 120% intensity resulted in significantly greater acute improvements in ROM, peak passive torque, and stiffness than stretching at 100% intensity. Four weeks of stretching increased ROM and peak passive torque but did not decrease passive stiffness, regardless of the stretching intensity.

Application of the novel estimation method by shear wave elastography using vibrator to human skeletal muscle.

In recent years, non-invasive measurement of tissue stiffness (hardness) using ultrasound elastography has attracted considerable attention. It has been used to evaluate muscle stiffness in the fields of rehabilitation, sports, and orthopedics. However, ultrasonic diagnostic devices with elastography systems are expensive and clinical use of such devices has been limited. In this study, we proposed a novel estimation method for vibration-based shear wave elastography measurement of human skeletal muscle, then determined its reproducibility and reliability. The coefficient of variation and correlation coefficient were used to determine reproducibility and reliability of the method by measuring the shear wave velocities in konjac phantom gels and agar phantom gels, as well as skeletal muscle. The intra-day, day-to-day, and inter-operator reliabilities were good when measuring the shear wave velocities in phantom gels. The intra-day and day-to-day reliabilities were good when measuring the shear wave velocities in skeletal muscle. The findings confirmed adequate reproducibility and reliability of the novel estimation method for vibration-based shear wave elastography. Therefore, the proposed measurement method may be a useful tool for evaluation of muscle stiffness.

The protective role of localized nitric oxide production during inflammation may be mediated by the heme oxygenase-1/carbon monoxide pathway.

Nitric oxide (NO) is an important part of the host defense mechanism; however, it displays both pro- and anti-inflammatory properties depending on its location and concentration. Importantly, excessive or inappropriate NO production can cause tissue damage. Systemic and local administration of NO synthase (NOS) inhibitors ameliorates and may exacerbate the inflammatory response, respectively. Here, we used a carrageenan-induced pleurisy model of acute inflammation in rats to confirm the location-dependent effects of NO and investigate the underlying mechanisms. As expected, localized suppression of NO production exacerbated inflammation, as evidenced by increased pleural exudate volumes and leukocyte counts and enhanced activity of enzymes related to oxidative stress. In contrast, local NO supplementation reduced leukocyte infiltration, vascular permeability, and the activity of oxidative stress-related enzymes. Interestingly, inhibition of heme oxygenase-1 (HO-1) reversed the anti-inflammatory effects of localized NO production, while the addition of hemin (HO-1 substrate) or carbon monoxide (CO; HO-1 metabolite) decreased leukocyte migration and exudation. Together, these findings confirm a protective role for NO at the inflammatory site, which appears to be mediated via NOS induction of the HO-1/CO pathway. Thus, NO supplementation may be a potential new treatment for oxidative stress-associated inflammatory diseases.

Changes in Flexibility and Force are not Different after Static Versus Dynamic Stretching.

In this study, we examined the effects of static and dynamic stretching on range of motion (ROM), passive torque (PT) at pain onset, passive stiffness, and isometric muscle force. We conducted a randomized crossover trial in which 16 healthy young men performed a total of 300 s of active static or dynamic stretching of the right knee flexors on two separate days in random order. To assess the effects of stretching, we measured the ROM, PT at pain onset, passive stiffness during passive knee extension, and maximum voluntary isometric knee flexion force using an isokinetic dynamometer immediately before and after stretching. Both static and dynamic stretching significantly increased the ROM and PT at pain onset (p<0.01) and significantly decreased the passive stiffness and isometric knee flexion force immediately after stretching (p<0.01). However, the magnitude of change did not differ between the two stretching methods for any measurements. Our results suggest that 300 s of either static or dynamic stretching can increase flexibility and decrease isometric muscle force; however, the effects of stretching do not appear to differ between the two stretching methods.

Dynamic Stretching Has Sustained Effects on Range of Motion and Passive Stiffness of the Hamstring Muscles.

Dynamic stretching (DS) is often performed during warm-up to help avoid hamstring muscle injuries, increase joint flexibility, and optimize performance. We examined the effects of DS of the hamstring muscles on passive knee extension range of motion (ROM), passive torque (PT) at the onset of pain (as a measure of stretch tolerance), and passive stiffness of the muscle-tendon unit over an extended period after stretching. Twenty-four healthy subjects participated, with 12 each in the experimental and control groups. Stretching was performed, and measurements were recorded using an isokinetic dynamometer pre-intervention, and at 0, 15, 30, 45, 60, 75, and 90 min post-intervention. DS consisted of ten 30-s sets of 15 repetitions of extension and relaxation of the hamstrings. ROM increased significantly (range, 7%-10%) immediately after DS, and the increase was sustained over 90 min. PT at the onset of pain also increased immediately by 10% but returned to baseline by 30 min. Passive stiffness decreased significantly (range, 7.9%-16.7%) immediately after DS, and the decrease was sustained over 90 min. Post-DS values were normalized to pre-DS values for the respective outcomes in both groups. ROM was significantly higher (range, 7.4%-10%) and passive stiffness was significantly lower (range, 5.4%-14.9%) in the experimental group relative to the control group at all time points. Normalized PT values at the onset of pain were significantly higher in the experimental group at 0-15 min than in the controls, but the differences were smaller at 30-45 min and not significant thereafter. We conclude that DS increases ROM and decreases passive stiffness in a sustained manner, and increases PT at the onset of pain for a shorter period. Overall, our results indicate that when performed prior to exercise, DS is beneficial for the hamstring muscles in terms of increasing flexibility and reducing stiffness.

Hamstring Stiffness Returns More Rapidly After Static Stretching Than Range of Motion, Stretch Tolerance, and Isometric Peak Torque.

Context: Hamstring injuries are common, and lack of hamstring flexibility may predispose to injury. Static stretching not only increases range of motion (ROM) but also results in reduced muscle strength after stretching. The effects of stretching on the hamstring muscles and the duration of these effects remain unclear. Objective: To determine the effects of static stretching on the hamstrings and the duration of these effects. Design: Randomized crossover study. Setting: University laboratory. Participants: A total of 24 healthy volunteers. Interventions: The torque-angle relationship (ROM, passive torque [PT] at the onset of pain, and passive stiffness) and isometric muscle force using an isokinetic dynamometer were measured. After a 60-minute rest, the ROM of the dynamometer was set at the maximum tolerable intensity; this position was maintained for 300 seconds, while static PT was measured continuously. The torque-angle relationship and isometric muscle force after rest periods of 10, 20, and 30 minutes were remeasured. Main Outcome Measures: Change in static PT during stretching and changes in ROM, PT at the onset of pain, passive stiffness, and isometric muscle force before stretching were compared with 10, 20, and 30 minutes after stretching. Results: Static PT decreased significantly during stretching. Passive stiffness decreased significantly 10 and 20 minutes after stretching, but there was no significant prestretching versus poststretching difference after 30 minutes. PT at the onset of pain and ROM increased significantly after stretching at all rest intervals, while isometric muscle force decreased significantly after all rest intervals. Conclusions: The effect of static stretching on passive stiffness of the hamstrings was not maintained as long as the changes in ROM, stretch tolerance, and isometric muscle force. Therefore, frequent stretching is necessary to improve the viscoelasticity of the muscle-tendon unit. Muscle force decreased for 30 minutes after stretching; this should be considered prior to activities requiring maximal muscle strength.

Inhibition of ubiquitin-activating enzyme protects against organ injury after intestinal ischemia-reperfusion.

Intestinal ischemia-reperfusion (I/R) occurs in various clinical settings, such as transplantation, acute mesenteric arterial occlusion, trauma, and shock. I/R injury causes severe systemic inflammation, leading to multiple organ dysfunction associated with high mortality. The ubiquitin proteasome pathway has been indicated in the regulation of inflammation, particularly through the NF-κB signaling pathway. PYR-41 is a small molecular compound that selectively inhibits ubiquitin-activating enzyme E1. A mouse model of intestinal I/R injury by clamping the superior mesenteric artery for 45 min was performed to evaluate the effect of PYR-41 treatment on organ injury and inflammation. PYR-41 was administered intravenously at the beginning of reperfusion. Blood and organ tissues were harvested at 4 h after reperfusion. PYR-41 treatment improved the morphological structure of gut and lung after I/R, as judged by hematoxylin and eosin staining. It also reduced the number of apoptotic terminal deoxynucleotidyl transferase dUTP nick end-labeling-positive cells and caspase-3 activity in the organs. PYR-41 treatment decreased the expression of proinflammatory cytokines IL-6 and IL-1ß as well as chemokines keratinocyte chemoattractant and macrophage inflammatory protein-2 in the gut and lung, which leads to inhibition of neutrophils infiltrating into these organs. The serum levels of IL-6, aspartate aminotransferase, and lactate dehydrogenase were reduced by the treatment. The IκB degradation in the gut increased after I/R was inhibited by PYR-41 treatment. Thus, ubiquitination may be a potential therapeutic target for treating patients suffering from intestinal I/R. NEW & NOTEWORTHY Excessive inflammation contributes to organ injury from intestinal ischemia-reperfusion (I/R) in many clinical conditions. NF-κB signaling is very important in regulating inflammatory response. In an experimental model of gut I/R injury, we demonstrate that administration of a pharmacological inhibitor of ubiquitination process attenuates NF-κB activation, leading to reduction of inflammation, tissue damage, and apoptosis in the gut and lungs. Therefore, ubiquitination process may serve as a therapeutic target for treating patients with intestinal I/R injury.

PYR-41, A Ubiquitin-Activating Enzyme E1 Inhibitor, Attenuates Lung Injury in Sepsis.

During sepsis, systemic inflammation is observed and is associated with multiple organ failure. Activation of NF-κB is crucial for inducing inflammation, which is controlled by degradation of inhibitor molecules (IκB). The ubiquitination proteasome pathway is responsible for the regulation of protein turnover. In this study, we hypothesized that administration of 4[4-(5-nitro-furan-2-ylmethylene)-3, -dioxo-pyrazolidin-1-yl]-benzoic acid ethyl ester (PYR-41), an inhibitor of ubiquitination, could reduce inflammation and organ injury in septic mice. PYR-41 prevented the reduction of IκB protein levels and inhibited release of tumor necrosis factor (TNF)-α in mouse macrophage RAW264.7 cells at 4 h after lipopolysaccharide stimulation dose-dependently. Male C57BL/6 mice were subjected to cecal ligation and puncture (CLP) to induce sepsis. PYR-41 (5 mg/kg) or dimethyl sulfoxide in saline (vehicle) was injected intravenously immediately after CLP. At 20 h after CLP, PYR-41 treatment significantly decreased serum levels of proinflammatory cytokines (TNF-α, interleukin [IL]-1ß, and IL-6) and organ injury markers (aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase). PYR-41 significantly improved microscopic structure, and reduced myeloperoxidase activity, number of apoptotic cells and caspase-3 degradation in the lungs of septic mice. The reduced protein levels of IκB in the lungs after CLP were restored by PYR-41 treatment. PYR-41 inhibited the expression of cytokines (IL-1ß and IL-6), chemokines (keratinocyte-derived chemokine and macrophage inflammatory protein 2), and inflammatory mediators (cyclooxygenase-2 and inducible nitric oxide synthase) in the lungs of septic mice. Importantly, PYR-41 significantly increased 10-day survival in septic mice from 42% to 83%. Therefore, targeting ubiquitination by PYR-41 to inhibit NF-κB activation may represent a potential strategy of sepsis therapeutics.

Metabolic engineering of Synechococcus elongatus PCC 7942 for improvement of 1,3-propanediol and glycerol production based on in silico simulation of metabolic flux distribution.

BACKGROUND: Production directly from carbon dioxide by engineered cyanobacteria is one of the promising technologies for sustainable future. Previously, we have successfully achieved 1,3-propanediol (1,3-PDO) production using Synechococcus elongatus PCC 7942 with a synthetic metabolic pathway. The strain into which the synthetic metabolic pathway was introduced produced 3.48 mM (0.265 g/L) 1,3-PDO and 14.3 mM (1.32 g/L) glycerol during 20 days of incubation. In this study, the productivities of 1,3-PDO were improved by gene disruption selected by screening with in silico simulation. METHODS: First, a stoichiometric metabolic model was applied to prediction of cellular metabolic flux distribution in a 1,3-PDO-producing strain of S. elongatus PCC 7942. A genome-scale model of S. elongatus PCC 7942 constructed by Knoop was modified by the addition of a synthetic metabolic pathway for 1,3-PDO production. Next, the metabolic flux distribution predicted by metabolic flux balance analysis (FBA) was used for in silico simulation of gene disruption. As a result of gene disruption simulation, NADPH dehydrogenase 1 (NDH-1) complexes were found by screening to be the most promising candidates for disruption to improve 1,3-PDO production. The effect of disruption of the gene encoding a subunit of the NDH-1 complex was evaluated in the 1,3-PDO-producing strain. RESULTS AND CONCLUSIONS: During 20 days of incubation, the ndhF1-null 1,3-PDO-producing strain showed the highest titers: 4.44 mM (0.338 g/L) 1,3-PDO and 30.3 mM (2.79 g/L) glycerol. In this study, we successfully improved 1,3-PDO productivity on the basis of in silico simulation of gene disruption.

Acute Effects of the Different Intensity of Static Stretching on Flexibility and Isometric Muscle Force.

Kataura, S, Suzuki, S, Matsuo, S, Hatano, G, Iwata, M, Yokoi, K, Tsuchida, W, Banno, Y, and Asai, Y. Acute effects of the different intensity of static stretching on flexibility and isometric muscle force. J Strength Cond Res 31(12): 3403-3410, 2017-In various fields, static stretching is commonly performed to improve flexibility, whereas the acute effects of different stretch intensities are unclear. Therefore, we investigated the acute effects of different stretch intensities on flexibility and muscle force. Eighteen healthy participants (9 men and 9 women) performed 180-second static stretches of the right hamstrings at 80, 100, and 120% of maximum tolerable intensity without stretching pain, in random order. The following outcomes were assessed as markers of lower limb function and flexibility: static passive torque (SPT), range of motion (ROM), passive joint (muscle-tendon) stiffness, passive torque (PT) at onset of pain, and isometric muscle force. Static passive torque was significantly decreased after all stretching intensities (p ≤ 0.05). Compared with before stretching at 100 and 120% intensities, ROM and PT were significantly increased after stretching (p ≤ 0.05), and passive stiffness (p = 0.05) and isometric muscle force (p ≤ 0.05) were significantly decreased. In addition, ROM was significantly greater after stretching at 100 and 120% than at 80%, and passive stiffness was significantly lower after 120% than after 80% (p ≤ 0.05). However, all measurements except SPT were unchanged after 80% intensity. There was a weak positive correlation between the intensities of stretching and the relative change for SPT (p ≤ 0.05), a moderate positive correlation with ROM (p ≤ 0.05), and a moderate positive correlation with passive stiffness (p ≤ 0.05). These results indicate that static stretching at greater intensity is more effective for increasing ROM and decreasing passive muscle-tendon stiffness.

Heat Stress Modulates Both Anabolic and Catabolic Signaling Pathways Preventing Dexamethasone-Induced Muscle Atrophy In Vitro.

It is generally recognized that synthetic glucocorticoids induce skeletal muscle weakness, and endogenous glucocorticoid levels increase in patients with muscle atrophy. It is reported that heat stress attenuates glucocorticoid-induced muscle atrophy; however, the mechanisms involved are unknown. Therefore, we examined the mechanisms underlying the effects of heat stress against glucocorticoid-induced muscle atrophy using C2C12 myotubes in vitro, focusing on expression of key molecules and signaling pathways involved in regulating protein synthesis and degradation. The synthetic glucocorticoid dexamethasone decreased myotube diameter and protein content, and heat stress prevented the morphological and biochemical glucocorticoid effects. Heat stress also attenuated increases in mRNAs of regulated in development and DNA damage responses 1 (REDD1) and Kruppel-like factor 15 (KLF15). Heat stress recovered the dexamethasone-induced inhibition of PI3K/Akt signaling. These data suggest that changes in anabolic and catabolic signals are involved in heat stress-induced protection against glucocorticoid-induced muscle atrophy. These results have a potentially broad clinical impact because elevated glucocorticoid levels are implicated in a wide range of diseases associated with muscle wasting. J. Cell. Physiol. 232: 650-664, 2017. © 2016 The Authors. Journal of Cellular Physiology published by Wiley Periodicals, Inc.

Milk fat globule epidermal growth factor-factor 8-derived peptide attenuates organ injury and improves survival in sepsis.

INTRODUCTION: Sepsis involves overwhelming inflammatory responses with subsequent immune-suppression that can lead to multiple organ dysfunction and ultimately death. Milk fat globule epidermal growth factor-factor 8 (MFG-E8) is a secretory protein found to have multiple biological activities against autoimmune and inflammatory diseases. MFG-E8 contains an Arg-Gly-Asp (RGD) motif involved in cell-cell and cell-matrix interactions. In sepsis, excessive neutrophils migration through endothelial cells and matrix to sites of inflammation results in organ damage. We hypothesized that MFG-E8-derived short peptides (MSP) flanking its RGD motif could provide protection against organ injury in sepsis. METHODS: The differentiated human neutrophil-like HL-60 cells (dHL60) were incubated with a series of peptides flanking the RGD motif of human MFG-E8 for a cell adhesion assay to fibronectin or human pulmonary artery endothelial cells (PAECs). For the induction of sepsis, male C57BL/6 mice (20-25 g) were subjected to cecal ligation and puncture (CLP). Peptide MSP68 (1 mg/kg body weight) or normal saline (vehicle) was injected intravenously at 2 h after CLP. Blood and tissue samples were collected at 20 h after CLP for various measurements. RESULTS: After screening, peptide MSP68 (VRGDV) had the highest inhibition of dHL-60 cell adhesion to fibronectin by 55.8 % and to PAEC by 67.7 %. MSP68 treatment significantly decreased plasma levels of organ injury marker AST by 37.1 % and the proinflammatory cytokines IL-6 and TNF-α by 61.9 % and 22.1 %, respectively after CLP. MSP68 improved the integrity of microscopic architectures, decreased IL-6 levels in the lungs by 85.1 %, and reduced apoptosis. MSP68 treatment also significantly reduced the total number of neutrophil infiltration by 61.9 % and 48.3 % as well as MPO activity by 40.8 % and 47.3 % in the lungs and liver, respectively, after CLP. Moreover, the number of bacteria translocated to mesenteric lymph nodes was decreased by 57 % with MSP68 treatment. Finally, the 10-day survival rate was increased from 26 % in the vehicle group to 58 % in the MSP68-treated group. CONCLUSIONS: MSP68 effectively inhibits excessive neutrophils infiltrating to organs, leading to moderate attenuation of organ injury and significantly improved survival in septic mice. Thus, MSP68 may be a potential therapeutic agent for treating sepsis.

Predictive factors for the outcome of multidisciplinary treatments in chronic low back pain at the first multidisciplinary pain center of Japan.

[Purpose] Multidisciplinary treatments are recommended for treatment of chronic low back pain. The aim of this study was to show the associations among multidisciplinary treatment outcomes, pretreatment psychological factors, self-reported pain levels, and history of pain in chronic low back pain patients. [Subjects and Methods] A total of 221 chronic low back pain patients were chosen for the study. The pretreatment scores for the 10-cm Visual Analogue Scale, Hospital Anxiety and Depression Scale, Pain Catastrophizing Scale, Short-Form McGill Pain Questionnaire, Pain Disability Assessment Scale, pain drawings, and history of pain were collected. The patients were divided into two treatment outcome groups a year later: a good outcome group and a poor outcome group. [Results] One-hundred eighteen patients were allocated to the good outcome group. The scores for the Visual Analogue Scale, Pain Disability Assessment Scale, and affective subscale of the Short-Form McGill Pain Questionnaire and number of nonorganic pain drawings in the good outcome group were significantly lower than those in the poor outcome group. Duration of pain in the good outcome group was significantly shorter than in the poor outcome group. [Conclusion] These findings help better predict the efficacy of multidisciplinary treatments in chronic low back pain patients.

Differential alterations of tissue T-cell subsets after sepsis.

Among immune cells in responding to sepsis, macrophages and neutrophils have been extensively studied, while the contribution of T lymphocytes and natural killer T (NKT) cells is less well characterized. Here we monitored tissue specific changes of T cell subsets in male C57BL/6 mice subjected to sham operation or cecal ligation and puncture (CLP) to induce polymicrobial sepsis. Thymus, spleen, liver, lungs and blood were processed and analyzed 20h later. Total lymphocyte count showed a significant reduction in septic thymus, spleen and blood but not in lungs and liver. The septic thymi were hypocellular with severe reduction in cell numbers of immature CD4(+)CD8(+) subset. CD4(+) T and CD8(+) T lymphocyte numbers in septic spleens were also significantly reduced, but the frequency of CD4(+)CD25(+) Tregs was significantly increased. In addition, naïve and Tcm CD4(+) T cell numbers were significantly reduced in the septic spleens. By contrast, in septic liver the CD8(+) T cell numbers were significantly increased, whereas NKT cell numbers were reduced, but more activated with increased CD69 and CD25 expression. In the septic lungs, the CD4(+) T and CD8(+) T cell numbers showed no significant change, whereas they were severely reduced in the septic blood. Overall, this study provides important information on the alterations of different T-cell subsets in various tissues after sepsis.

MFG-E8 inhibits neutrophil migration through αvß3-integrin-dependent MAP kinase activation.

We have previously demonstrated the involvement of milk fat globule-epidermal growth factor-factor 8 (MFG­E8) in reducing neutrophil infiltration in a murine model of acute lung injury (ALI). In the present study, we aimed to delineate the mechanisms through which MFG­E8 attenuates neutrophil migration. Recombinant human MFG­E8 (rhMFG­E8) was expressed and purified in our facility. The human differentiated neutrophil cell line, dHL­60, was treated with rhMFG­E8 and cell migration assay was performed in a Boyden chamber using recombinant interleukin­8 (IL­8) as the chemoattractant. Surface CXCR2 and intracellular G protein­coupled receptor kinase 2 (GRK2) levels were evaluated by flow cytometry or western blot analysis. The levels of mitogen­activated protein (MAP) kinases were determined by western blot analysis. Treatment with rhMFG­E8 resulted in a significant inhibition of dHL­60 cell migration in a dose­dependent manner. There was a 46% decrease in CXCR2 expression in the rhMFG­E8­treated dHL­60 cells, which was associated with a 32% increase in GRK2 expression. In the dHL­60 cells, treatment with rhMFG­E8 promoted the phosphorylation of p38 and extracellular signal-regulated kinase (ERK) within 10­30 min. The use of SB203580, a p38 inhibitor, and PD98059, an ERK inhibitor, resulted in the restoration of dHL­60 cell migration which was significantly inhibited treatment with rhMFG­E8. Furthermore, blocking the MFG­E8 receptors, αvß3/αvß5­integrins, by anti­αv­integrin neutralizing antibody (Ab) inhibited the activation of p38 and ERK, and reversed the rhMFG­E8­induced inhibition of dHL­60 cell migration. Finally, treatment of the dHL­60 cells with SB203580 and PD98059 neutralized the rhMFG­E8­induced downregulation of CXCR2 expression and upregulation of GRK2 expression, as well as the inhibitory effects on cell migration. Our findings reveal a novel mechanism of action of MFG­E8 through which it inhibits neutrophil migration through αvß3-integrin-dependent MAP kinase activation.

Changes in force and stiffness after static stretching of eccentrically-damaged hamstrings.

PURPOSE: This study compared responses to static stretching between eccentrically damaged and non-damaged muscles. METHODS: Twelve young men performed 60 maximum knee flexor eccentric contractions of one leg, and received a 300-s continuous passive static stretching at tolerable intensity without pain to both knee flexors at 2 and 4 days after the eccentric exercise. Range of motion (ROM) and passive stiffness during knee extension, passive torque at onset of pain (PT), maximum voluntary isometric (MVC-ISO) and isokinetic concentric contraction torque (MVC-CON), and visual analogue scale (VAS) for muscle soreness were measured before, immediately after, 60 min, 2 and 4 days after exercise as well as before, immediately after, 20 and 60 min after the stretching. Changes in these variables after eccentric exercise and stretching were compared between limbs. RESULTS: The eccentric exercise decreased MVC-ISO, MVC-CON, ROM and PT, and increased passive stiffness and VAS (p < 0.05), suggesting that muscle damage was induced to the knee flexors. ROM and PT increased after stretching for both limbs; however, the magnitude of the increase was greater (p < 0.05) for the damaged than non-damaged limb. Passive stiffness decreased for both limbs similarly (4-7 %) at immediately after stretching (p < 0.05). Significant decreases in MVC-ISO torque (7-11 %) after stretching were observed only for the non-damaged limb (p < 0.05), but MVC-CON torque did not change after stretching for both limbs. VAS decreased for the exercised limb after stretching (p < 0.05). CONCLUSIONS: These results suggest that the static stretching at tolerable intensity without pain produced greater positive effects on damaged than non-damaged muscles.