Distinct adaptations of muscle endurance but not strength or hypertrophy to low-load resistance training with and without blood flow restriction.

Low-load resistance training promotes muscle strength and hypertrophic adaptations when combined with blood flow restriction (BFR). However, the effect of BFR on muscle endurance remains unclear. The aim of this study was to clarify the effects of BFR on muscle performance and adaptation, with special reference to local muscle endurance. In experiment 1, eight healthy men performed unilateral elbow flexion exercise to failure at 30% of one-repetition maximum with BFR (at 40% of estimated arterial occlusion pressure) and free blood flow (FBF). During the exercise, muscle activity and tissue oxygenation were measured from the biceps brachii. In experiment 2, another eight healthy men completed 6 weeks of elbow flexion training with BFR and FBF. The number of repetitions to failure at submaximal load (Rmax), the estimated time for peak torque output to decay by 50% during repetitive maximum voluntary contractions (half-time), one-repetition maximum, isometric strength and muscle thickness of elbow flexors were measured pre- and post-training. Blood flow restriction resulted in fewer repetitions and lower muscle tissue oxygenation at the end of exercise than FBF, while the muscle activity increased similarly to repetition failure. Blood flow restriction also resulted in a smaller post-training Rmax, which was strongly correlated with the total exercise volume over the 6 week period. Despite the smaller exercise volume, BFR resulted in similar improvements in half-time, muscle strength and thickness compared with FBF. These results suggest that the application of BFR can attenuate muscle endurance adaptations to low-load resistance training by decreasing the number of repetitions during exercise, both acutely and chronically.

Alterations in neuromuscular junction morphology with ageing and endurance training modulate neuromuscular transmission and myofibre composition.

Both ageing and exercise training affect the neuromuscular junction (NMJ) structure. Morphological alterations in the NMJ have been considered to influence neuromuscular transmission and myofibre properties, but the direct link between the morphology and function has yet to be established. We measured the neuromuscular transmission, myofibre composition and NMJ structure of 5-month-old (young) and 24-month-old untrained (aged control) and trained (aged trained) mice. Aged trained mice were subjected to 2 months of endurance training before the measurement. Neuromuscular transmission was evaluated in vivo as the ratio of ankle plantar flexion torque evoked by the sciatic nerve stimulation to that by direct muscle stimulation. The torque ratio was significantly lower in aged mice than in young and aged trained mice at high-frequency stimulations, showing a significant positive correlation with voluntary grip strength. The degree of pre- to post-synaptic overlap of the NMJ was also significantly lower in aged mice and positively correlated with the torque ratio. We also found that the proportion of fast-twitch fibres in the soleus muscle decreased with age, and that age-related denervation occurred preferentially in fast-twitch fibres. Age-related denervation and a shift in myofibre composition were partially prevented by endurance training. These results suggest that age-related deterioration of the NMJ structure impairs neuromuscular transmission and alters myofibre composition, but these alterations can be prevented by structural amelioration of NMJ with endurance training. Our findings highlight the importance of the NMJ as a major determinant of age-related deterioration of skeletal muscles and the clinical significance of endurance training as a countermeasure. KEY POINTS: The neuromuscular junction (NMJ) plays an essential role in neuromuscular transmission and the maintenance of myofibre properties. We show that neuromuscular transmission is impaired with ageing but recovered by endurance training, which contributes to alterations in voluntary strength. Neuromuscular transmission is associated with the degree of pre- to post-synaptic overlap of the NMJ. Age-related denervation of fast-twitch fibres and a shift in myofibre composition toward a slower phenotype are partially prevented by endurance training. Our study provides substantial evidence that age-related and exercise-induced alterations in neuromuscular transmission and myofibre properties are associated with morphological changes in the NMJ.

Influence of temperature on twitch potentiation following submaximal voluntary contractions in human plantar flexor muscles.

This study aimed to clarify the influence of temperature on post-activation twitch potentiation, a possible indicator of fast-twitch fiber activation during a preceding voluntary contraction. Ten healthy males immersed their left lower leg in water of different temperatures (cold: 0°C, neutral: 32-35°C, hot: ~43°C) for 20 min each. In each temperature condition, they performed submaximal (10%-50% of maximal voluntary contraction torque measured before water immersion) and maximal plantar flexions. Immediately after each voluntary contraction, twitch contractions were evoked with supramaximal stimulation of the posterior tibial nerve. The magnitude of twitch potentiation, defined as a percent increase in twitch torque following a voluntary contraction, increased with the intensity of the preceding voluntary contraction. The magnitude of twitch potentiation after the maximal voluntary contraction was smaller in Cold than in the other temperature conditions. However, temperature had no influence on the relative magnitude of twitch potentiation following the submaximal contractions. In addition, there was no difference in electromyographic activity between the gastrocnemius and soleus muscles in any temperature conditions. Collectively, the temperature dependence was not observed when using twitch potentiation or electromyographic amplitude as an indicator of fast-twitch fiber activation during brief submaximal voluntary contractions.

Synthesis of novobiocin derivatives and evaluation of their antigonococcal activity and pharmacokinetics.

Gonorrhea has become a serious problem because the number of infected people is increasing and the multi-drug resistance of the causative bacteria, Neisseria gonorrhoeae, is progressing. To develop novel drugs against resistant N. gonorrhoeae, we focused on the antibiotic novobiocin (1). This natural product has a different mechanism of action from existing drugs for gonorrhea, which may make it effective against resistant strains. Actually, it was applied to resistant N. gonorrhoeae, and moderate antibacterial activity was confirmed. Based on this result, we investigated the development of an antigonococcal drug with 1 as the lead compound. The pharmacophore is thought to be the noviose sugar moiety, especially around the 3'-position, so we derivatized this part in order to improve antibacterial activity. As a result, we found that 5 with an methylpyrrole ester structure have a very potent antibacterial activity. This derivative also showed excellent antigonococcal activity against resistant strains in vitro, however it has poor water solubility and pharmacokinetics because it is the acidic lipid-soluble compound. Therefore, we considered introduction of a basic substituent into the molecule would result in an amphoteric compound with improved water solubility, and we investigated further derivatization. As a result of synthesizing various derivatives, we found 47 containing imidazole with strong antigonococcal activity and greatly improved water solubility. This derivative has also improved metabolism and blood concentration in vivo, and is expected to be orally absorbed. Based on these results, we believe that 47 is a very promising anti-gonococcal lead compound and has great potential for further development.

Age and sex differences in force steadiness and intermuscular coherence of lower leg muscles during isometric plantar flexion.

Age- and sex-related alterations in the control of multiple muscles during contractions are not well understood. The purpose of the present study was to examine the age and sex differences in force steadiness and intermuscular coherence (IMC), and thereby to clarify the functional role of IMC during plantar flexion. Twenty-six young (YNG, 23-34 years), thirty middle-aged (MID, 35-64 years) and twenty-four older adults (OLD, 65-82 years) performed submaximal isometric contractions of plantar flexion, while electromyography was recorded from the soleus (SOL), gastrocnemius lateralis/medialis (GL/GM) and tibialis anterior (TA) muscles. Coefficient of variation (CV) of torque and IMC in the alpha, beta and gamma bands was calculated. We found that OLD demonstrated significantly higher torque CV than YNG and MID, and males demonstrated significantly higher torque CV than females (both p < 0.05). The IMC in the gamma band (five out of the six pairs) was significantly higher in YNG than MID and/or OLD (p < 0.05), while the gamma band IMC between GL and SOL was significantly higher in females. However, age or sex differences were not detected in the alpha or beta band. Moreover, the gamma band IMC between SOL and TA had a weak (r = - 0.229) but significant (p < 0.05) negative correlation with torque CV. These results suggest that force steadiness differs with age and sex, and that the higher gamma band IMC may contribute to more stable force control during plantar flexion.

Higher Training Frequency Is Important for Gaining Muscular Strength Under Volume-Matched Training.

Background: This study investigated the effect of volume-matched strength training programs with different frequency and subsequent detraining on muscle size and strength. Methods: During a training period of 11 weeks, untrained subjects (age: 22.3 ± 0.9 years, height: 173.1 ± 4.8 cm and body mass: 66.8 ± 8.4 kg) performed knee-extension exercise at 67% of their estimated one-repetition maximum either one session per week (T1 group: 6 sets of 12 repetitions per session; n = 10) or three sessions per week (T3 group: 2 sets of 12 repetitions per session; n = 10). Rating of perceived exertion (RPE) and muscle stiffness were measured as an index of muscle fatigue and muscle damage, respectively. The magnitude of muscle hypertrophy was assessed with thigh circumference and the quadriceps muscle thickness. The changes in muscle strength were measured with isometric maximum voluntary contraction torque (MVC). Results: During the training period, RPE was significantly higher in the T1 than in the T3 (p < 0.001). After 11 weeks of training, both groups exhibited significant improvements in thigh circumference, muscle thickness, and MVC compared with baseline values. However, there was a significant group difference in MVC improvement at week 11 (T1: 43.5 ± 15.5%, T3: 65.2 ± 23.2%, p < 0.05). After 6 weeks of detraining, both groups showed the significant decreases in thigh circumference and muscle thickness from those at the end of training period, while no significant effect of detraining was observed in MVC. Conclusion: These results suggest that three training sessions per week with two sets are recommended for untrained subjects to improve muscle strength while minimizing fatigue compared to one session per week with six sets.

Acute effect of wearing compression stockings on lower leg swelling and muscle stiffness in healthy young women.

Compression stockings are known to be effective in reducing peripheral oedema or leg swelling by increasing tissue pressure and venous blood flow. While previous studies on compression stockings have focused on its prolonged, preventive effect on leg swelling, the aim of this study was to investigate an acute effect of wearing compression stockings on lower leg swelling and muscle stiffness. Twenty healthy women aged 18-23 years participated in the experiment conducted in the evening, in which they wore below-knee graduated compression stockings and rested in a seated position for 30 min. Before and after the application of stockings, maximum calf circumference, volume, extracellular water resistance (RECW ) and muscle stiffness of the right lower leg were determined by tape measure, water displacement volumetry, segmental bioelectrical impedance spectroscopy and ultrasound shear-wave elastography, respectively. The maximum calf circumference and the reciprocal of RECW (an index of extracellular fluid volume) significantly decreased after the application of stockings, whereas the total lower leg volume and the stiffness (shear modulus) of the medial gastrocnemius muscle tended to decrease. These changes, except for that in the medial gastrocnemius muscle stiffness, were opposite to those from morning to evening studied in the subgroup of participants (n = 8). However, partial correlation analysis failed to detect significant associations among these changes. These results suggest that even for a short period of application, compression stockings have some positive effects against lower leg swelling.

Synthesis and antibacterial activity of 4″ or 6″-alkanoylamino derivatives of arbekacin.

Arbekacin, an aminoglycoside antibiotic, is an important drug because it shows a potent efficacy against methicillin-resistant Staphylococcus aureus. However, resistance to arbekacin, which is caused mainly by the bifunctional aminoglycoside-modifying enzyme, has been observed, becoming a serious problem in medical practice. To create new arbekacin derivatives active against resistant bacteria, we modified the C-4″ and 6″ positions of its 3-aminosugar portion. Regioselective amination of the 6″-position gave 6″-amino-6″-deoxyarbekacin (1), and it was converted to a variety of 6″-N-alkanoyl derivatives (6a-z). Furthermore, regioselective modifications of the 4″-hydroxyl group were performed to give 4″-deoxy-4″-epiaminoarbekacin (2) and its 4″-N-alkanoyl derivatives (12 and 13). Their antibacterial activity against S. aureus, including arbekacin-resistant bacteria, was evaluated. It was observed that 6″-amino-6″-N-[(S)-4-amino-2-hydroxybutyryl]-6″-deoxyarbekacin (6o) showed excellent antibacterial activity, even better than arbekacin.

Length-force characteristics of in vivo human muscle reflected by supersonic shear imaging.

Recently, an ultrasound-based elastography technique has been used to measure stiffness (shear modulus) of an active human muscle along the axis of contraction. Using this technique, we explored 1) whether muscle shear modulus, like muscle force, is length dependent; and 2) whether the length dependence of muscle shear modulus is consistent between electrically elicited and voluntary contractions. From nine healthy participants, ankle joint torque and shear modulus of the tibialis anterior muscle were measured at five different ankle joint angles during tetanic contractions and during maximal voluntary contractions. Fascicle length, pennation angle, and tendon moment arm length of the tetanized tibialis anterior calculated from ultrasound images were used to reveal the length-dependent changes in muscle force and shear modulus. Over the range of joint angles examined, both force and shear modulus of the tetanized muscle increased with increasing fascicle length. Regression analysis of normalized data revealed a significant linear relationship between force and shear modulus (R(2) = 0.52, n = 45, P < 0.001). Although the length dependence of shear modulus was consistent, irrespective of contraction mode, the slope of length-shear modulus relationship was steeper during maximal voluntary contractions than during tetanic contractions. These results provide novel evidence that length-force relationship, one of the most fundamental characteristics of muscle, can be inferred from in vivo imaging of shear modulus in the tibialis anterior muscle. Furthermore, the estimation of length-force relationship may be applicable to voluntary contractions in which neural and mechanical interactions of multiple muscles are involved.

Amycolamicin: a novel broad-spectrum antibiotic inhibiting bacterial topoisomerase.

The abuse of antibacterial drugs imposes a selection pressure on bacteria that has driven the evolution of multidrug resistance in many pathogens. Our efforts to discover novel classes of antibiotics to combat these pathogens resulted in the discovery of amycolamicin (AMM). The absolute structure of AMM was determined by NMR spectroscopy, X-ray analysis, chemical degradation, and modification of its functional groups. AMM consists of trans-decalin, tetramic acid, two unusual sugars (amycolose and amykitanose), and dichloropyrrole carboxylic acid. The pyranose ring named as amykitanose undergoes anomerization in methanol. AMM is a potent and broad-spectrum antibiotic against Gram-positive pathogenic bacteria by inhibiting DNA gyrase and bacterial topoisomerase IV. The target of AMM has been proved to be the DNA gyrase B subunit and its binding mode to DNA gyrase is different from those of novobiocin and coumermycin, the known DNA gyrase inhibitors.

Activation of fast-twitch fibers assessed with twitch potentiation.

INTRODUCTION: The augmentation of twitch response following brief muscle activation, called twitch potentiation, has been shown to be much more pronounced in fast-twitch than in slow-twitch fibers. We thus explored the possibility of twitch potentiation as a noninvasive measure of fast-twitch fiber activation, by studying its dependence on the intensity of preceding contraction. METHODS: Twitch contraction of plantar flexor muscles was evoked with supramaximal stimulation of the posterior tibial nerve, before and immediately after 6-s voluntary contractions at intensities of 10-100% of maximal voluntary contraction (MVC). RESULTS: Except for low-intensity contractions (< 30%MVC), voluntary contraction induced twitch potentiation, the magnitude of which increased with increasing contraction intensity (P < 0.001). The shortened contractile process was associated with the potentiation. CONCLUSIONS: These results are consistent with the concept of "hierarchical order of fiber activation", suggesting that the magnitude of twitch potentiation reflects the activation of fast-twitch fibers during a brief contraction.

Acceleration and force reveal different mechanisms of electromechanical delay.

INTRODUCTION: Electromechanical delay (EMD) represents a series of complex processes of converting an electrical stimulus to a mechanical response. To quantify the contribution of electrochemical and mechanical processes of EMD in the human biceps brachii muscle over a wide range of elbow joint angles, we determined the onset of muscle contraction and the beginning of force development by recording acceleration of skin surface over the muscle and elbow flexion force, respectively. METHODS: Ten healthy male volunteers underwent two experimental sessions, in which submaximal paired-pulse stimuli were applied percutaneously to the resting biceps brachii muscle at 10 different elbow joint angles from 40° to 130° (0° represents full extension). RESULTS: The electrical stimulation induced repeatable contractions, in which the test-retest reliability of time parameters was sufficiently high (intraclass correlation coefficient=0.84-0.88). The time for electrochemical process ranged between 3.1±0.8 and 3.6±0.9 ms and was independent of elbow joint angle (P=0.64). The time for mechanical process and the total duration of EMD, however, were significantly greater at elbow flexion positions than at 40°, the most extended position in this study (P<0.05). Regression analysis revealed that at elbow flexion positions, the time for mechanical process increased significantly with decreasing the muscle-tendon length of the biceps brachii calculated from a musculoskeletal model (R=0.54, P<0.001). CONCLUSIONS: These results suggest that, in the human biceps brachii muscle, the prolongation of EMD at short muscle-tendon length is not attributed to the impairment of the electrochemical process of muscle contraction but to the increased slack within the muscle-tendon unit.

Unloaded shortening velocity of voluntarily and electrically activated human dorsiflexor muscles in vivo.

We have previously shown that unloaded shortening velocity (V(0)) of human plantar flexors can be determined in vivo, by applying the "slack test" to submaximal voluntary contractions (J Physiol 567:1047-1056, 2005). In the present study, to investigate the effect of motor unit recruitment pattern on V(0) of human muscle, we modified the slack test and applied this method to both voluntary and electrically elicited contractions of dorsiflexors. A series of quick releases (i.e., rapid ankle joint rotation driven by an electrical dynamometer) was applied to voluntarily activated dorsiflexor muscles at three different contraction intensities (15, 50, and 85% of maximal voluntary contraction; MVC). The quick-release trials were also performed on electrically activated dorsiflexor muscles, in which three stimulus conditions were used: submaximal (equal to 15%MVC) 50-Hz stimulation, supramaximal 50-Hz stimulation, and supramaximal 20-Hz stimulation. Modification of the slack test in vivo resulted in good reproducibility of V(0), with an intraclass correlation coefficient of 0.87 (95% confidence interval: 0.68-0.95). Regression analysis showed that V(0) of voluntarily activated dorsiflexor muscles significantly increased with increasing contraction intensity (R(2) = 0.52, P<0.001). By contrast, V(0) of electrically activated dorsiflexor muscles remained unchanged (R(2)<0.001, P = 0.98) among three different stimulus conditions showing a large variation of tetanic torque. These results suggest that the recruitment pattern of motor units, which is quite different between voluntary and electrically elicited contractions, plays an important role in determining shortening velocity of human skeletal muscle in vivo.

Synthesis and biological evaluation of dihydrofuran-fused perhydrophenanthrenes as a new anti-influenza agent having novel structural characteristic.

Dihydrofuran-fused perhydrophenanthrenes were synthesized by means of o-quinodimethane chemistry with high generality and stereoselectivity, and found to exhibit potent anti-influenza activity. These compounds exerted an inhibitory effect on various strains of influenza virus growth, including influenza A and B, with a concentration dependent manner, and direct cytotoxicity was low. Several biological experiments suggested that these new drugs affected a virus replication process before mRNA synthesis stage. Novel rigid cage-type of structural characteristic of the compounds has not been found in hitherto anti-influenza drugs, and will provide new basis and motif for exploring promising and unprecedented anti-influenza agents.

Shortening velocity of human triceps surae muscle measured with the slack test in vivo.

Unloaded shortening velocity (V(0)) of human triceps surae muscle was measured in vivo by applying the 'slack test', originally developed for determining V(0) of single muscle fibres, to voluntary contractions at varied activation levels (ALs). V(0) was measured from 10 subjects at five different ALs defined as a fraction (5, 10, 20, 40 and 60%) of the maximum voluntary contraction (MVC) torque. Although individual variability was apparent, V(0) tended to increase with AL (R(2) = 0.089; P = 0.035) up to 60%MVC (8.6 +/- 2.6 rad s(-1)). This value of V(0) at 60%MVC was comparable to the maximum shortening velocity of plantar flexors reported in the previous studies. Electromyographic analysis showed that the activities of soleus, medial gastrocnemius and lateral gastrocnemius muscles increased with AL during isometric contraction and after the application of quick release in a similar manner. Also, it showed that the activity of an antagonist, tibialis anterior muscle, was negligible, even though a slight increase took place after the quick release of agonist. Correlation analysis showed that there were no significant correlations between V(0) and MVC torque normalized with respect to body mass, although the correlation coefficient was relatively high at low ALs. The results suggest that in human muscle, V(0) represents the unloaded velocity of the fastest muscle fibres recruited, and increases with AL possibly because of progressive recruitment of faster fibres. Individual variability may be explained, at least partially, by the difference in fibre-type composition.

Synthesis of a new class of furan-fused tetracyclic compounds using o-quinodimethane chemistry and investigation of their antiviral Activity.

The synthesis and evaluation of antiviral activity of new furan-fused tetracyclic compounds are described. The syntheses were satisfactorily achieved on the basis of o-quinodimethane chemistry, using furan-containing benzocyclobutene derivatives as a substrate, in high generality and stereoselectivity. The various derivatives thus synthesized were examined on their inhibitory activity on virus growth using a hemagglutinin (HA) method, leading to a discovery of promising candidates for new antiviral drugs having high activity and good therapeutic index.