Aluminum naphthalocyanine conjugate as an MMP-2-activatable photoacoustic probe for in vivo tumor imaging.

Matrix metalloproteinase-2 (MMP-2), which is one of MMPs family, is known as an extracellular gelatinase controlling cancer cell adhesion, growth, and metastasis. Because of the great interest in MMP-2 activity, the detailed protocols for evaluating MMP-2-responsive contrast agents, especially photoacoustic probes for in vivo use, are helpful for researchers in the field. We here describe the detailed synthetic procedure of MMP-2-activatable photoacoustic probe AlNc-pep-PEG consisting of aluminum naphthalocyanine, MMP-2-responsive peptide sequence, and poly(ethylene glycol), which has recently been developed in our research group. The detailed measurement protocol of photoacoustic signal intensity in vitro and in vivo by using in-house built photoacoustic signal measurement system and photoacoustic imaging apparatus are also summarized.

MMP-2-Activatable Photoacoustic Tumor Imaging Probes Based on Al- and Si-Naphthalocyanines.

Enzyme-activatable photoacoustic probes are powerful contrast agents to visualize diseases in which a specific enzyme is overexpressed. In this study, aluminum and silicon naphthalocyanines (AlNc and SiNc, respectively) conjugated with matrix metalloprotease-2 (MMP-2)-responsive PLGLAG peptide sequence and poly(ethylene glycol) (PEG) as an axial ligand were designed and synthesized. AlNc-peptide-PEG conjugates AlNc-pep-PEG formed dimeric species interacting with each other through face-to-face H-aggregation in water, while SiNc-based conjugates SiNc-pep-PEG hardly interacted with each other because of the two bulky hydrophilic axial ligands. Both conjugates formed spherical nanometer-sized self-assemblies in water, generating photoacoustic waves under near-infrared photoirradiation. The treatment of MNc-peptide-PEG conjugates (M = Al, Si) with MMP-2 smoothly induced the cleavage of the PLGLAG sequence to release the hydrophilic PEG moiety, resulting in the aggregation of MNcs. By comparing the PA signal intensity changes at 680 and 760 nm, the photoacoustic signal intensity ratios were shown to be enhanced by 3-5 times after incubation with MMP-2. We demonstrated that MNc-peptide-PEG conjugates (M = Al, Si) could work as activatable photoacoustic probes in the in vitro experiment of MMP-2-overexpressed cell line HT-1080 as well as the in vivo photoacoustic imaging of HT-1080-bearing mice.

Relationship between resting medial gastrocnemius stiffness and drop jump performance.

Although the influence of the series elastic element of the muscle-tendon unit on jump performance has been investigated, the corresponding effect of the parallel elastic element remains unclear. This study examined the relationship between the resting calf muscle stiffness and drop jump performance. Twenty-four healthy men participated in this study. The shear moduli of the medial gastrocnemius and the soleus were measured at rest as an index of muscle stiffness using ultrasound shear wave elastography. The participants performed drop jumps from a 15 cm high box. The Spearman rank correlation coefficient was used to examine the relationships between shear moduli of the muscles and drop jump performance. The medial gastrocnemius shear modulus showed a significant correlation with the drop jump index (jump height/contact time) (r = 0.414, P = 0.044) and jump height (r = 0.411, P = 0.046), but not with contact time (P > 0.05). The soleus shear modulus did not correlate with these jump parameters (P > 0.05). These results suggest that the resting medial gastrocnemius stiffness can be considered as one of the factors that influence drop jump performance. Therefore, increase in resting muscle stiffness should enhance explosive athletic performance in training regimens.

Relationship Between Drop Jump Training-Induced Changes in Passive Plantar Flexor Stiffness and Explosive Performance.

Passive muscle stiffness is positively associated with explosive performance. Drop jump training may be a strategy to increase passive muscle stiffness in the lower limb muscles. Therefore, the purpose of this study was to examine the effect of 8-week drop jump training on the passive stiffness in the plantar flexor muscles and the association between training-induced changes in passive muscle stiffness and explosive performance. This study was a randomized controlled trial. Twenty-four healthy young men were divided into two groups, control and training. The participants in the training group performed drop jumps (five sets of 20 repetitions each) 3days per week for 8weeks. As an index of passive muscle stiffness, the shear moduli of the medial gastrocnemius and soleus were measured by shear wave elastography before and after the intervention. The participants performed maximal voluntary isometric plantar flexion at an ankle joint angle of 0° and maximal drop jumps from a 15cm high box. The rate of torque development during isometric contraction was calculated. The shear modulus of the medial gastrocnemius decreased for the training group (before: 13.5±2.1kPa, after: 10.6±2.1kPa); however, such a reduction was not observed in the control group. There was no significant group (control and training groups)×time (before and after the intervention) interaction for the shear modulus of the soleus. The drop jump performance for the training group improved, while the rate of torque development did not change. Relative changes in these measurements were not correlated with each other in the training group. These results suggest that drop jump training decreases the passive stiffness in the medial gastrocnemius, and training-induced improvement in explosive performance cannot be attributed to change in passive muscle stiffness.

Eight-Week Low-Intensity Squat Training at Slow Speed Simultaneously Improves Knee and Hip Flexion and Extension Strength.

Considering that the squat exercise requires flexion and extension of the knee and hip joints, a resistance training program based on squat exercises should efficiently increase the flexion and extension strength of both the knee and hip. To our knowledge, however, no study has simultaneously investigated the effects of squat training on both flexion and extension strength in both the knee and hip. Low-intensity squat exercises at slow speeds can be expected to effectively and safely improve knee and hip flexion and extension strength in a wide range of individuals. This study aimed to clarify whether knee and hip flexion and extension strength improved after an 8-week low-intensity squat training program at slow speed. Twenty-four untrained young men were randomly assigned to a training or control group. Participants in the training group performed 40% one-repetition maximum parallel squats at slow speed (4 s for concentric/eccentric actions), 3 days per week for 8 weeks. Before and after the intervention, isometric peak torque of the knee and hip flexors and extensors during maximal voluntary contraction (MVC) was determined. For the knee flexors and extensors, muscle volume was also measured. There were significant training-induced increases in peak torque (P < 0.05). The training effects on knee and hip extension torque (effect size = 0.36-0.38) were higher than those on knee and hip flexion torque (effect size = 0.09-0.13). The squat training used here increased both knee and hip flexion and extension strength, but the training effects on the flexion strength were less than those on the extension strength. Regarding the knee extensors, a significant training-related increase in muscle volume was found (P < 0.05) without neuromuscular adaptations. In addition, there were significant correlations between the training-induced increases in muscle volume and peak torque of KE. These results suggest that muscle hypertrophy may be responsible for increased muscle strength of the knee extensors after an 8-week low-intensity squat training program at slow speed.

Carbon dioxide hydrate as a recovery tool after fatigue of the plantar flexors.

This study investigated the effects of cooling the triceps surae with carbon dioxide hydrate (CDH), which is a gas hydrate, a crystalline structure belonging to the clathrates, on the recovery from muscle fatigue. Thirty-six healthy young men were equally and randomly assigned to an ICE group, a CDH group, or a non-cooling (NON) group. All participants performed 80 maximal voluntary isometric contractions (MVCs) of the plantar flexors as a fatiguing task. MVC torque and voluntary activation were determined before, immediately after, and 20 min after the fatiguing task. Evoked torque was similarly assessed except for immediately after the task. In the ICE and CDH groups, the triceps surae was cooled for 5 min using ice and CDH, starting 5 min after the fatiguing task, respectively. The MVC torque and voluntary activation were higher in order of before >20 min after >immediately after the fatiguing task regardless of group, and those time-course changes did not differ between the groups. A decrease in the evoked torque from before to 20 min after the fatiguing task was observed in the ICE and NON groups but not in the CDH group. These results suggest that cooling muscle with CDH can facilitate recovery from peripheral muscle fatigue. This may be due to an increase in blood flow caused by carbon dioxide contained within the CDH, and indicates the potential of CDH as a recovery tool after fatiguing exercise.

Active recovery has a positive and acute effect on recovery from fatigue induced by repeated maximal voluntary contractions of the plantar flexors.

This study investigated the acute effect of active recovery (AR) following fatigue induced by 80 three-second maximal voluntary isometric plantar flexion contractions (MVICs) in 12 young men. AR consisted of a total of 180 voluntary isometric ramp contractions of the plantar flexors (0.75-s contraction/relaxation) targeting 10% of MVIC torque. MVIC torque, voluntary activation and root mean square values of electromyographic signals for the triceps surae normalized by each peak-to-peak amplitude of compound motor action potential were determined before, and immediately, 10, 20 and 30 min after the fatiguing task. Evoked torques were similarly assessed except for immediately after it. The AR and passive recovery were randomly performed on two days by each participant between 5 min and 10 min after the fatiguing task. For all the parameters other than MVIC torque, there was no significant difference between the conditions at any time point. MVIC torque decreased significantly immediately after the fatiguing task regardless of condition (P < 0.05), and the corresponding decrease in MVIC torque recovered 30 min after the fatiguing task only in AR (P < 0.05). These results suggest an acute positive effect of AR on recovery of neuromuscular function and/or contractile properties after fatigue.