Muscle quality indices separately associate with joint-level power-related measures of the knee extensors in older males.

PURPOSE: The purpose of this study was to investigate associations of muscle quality indices with joint-level power-related measures in the knee extensors of thirty-two older males (65-88 years). METHODS: Muscle quality indices included: echo intensity, ratio of intracellular- to total water content (ICW/TW), and specific muscle strength. Echo intensity was acquired from the rectus femoris (EIRF) and vastus lateralis (EIVL) by ultrasonography. ICW/TW was computed from electrical resistance of the right thigh obtained by bioelectrical impedance spectroscopy. Specific muscle strength was determined as the normalized maximal voluntary isometric knee extension (MVIC) torque to estimated knee extensor volume. Isotonic maximal effort knee extensions with a load set to 20% MVIC torque were performed to obtain the knee extension power-related measures (peak power, rate of power development [RPD], and rate of velocity development [RVD]). Power and RPD were normalized to MVIC. RESULTS: There were no significant correlations between muscle quality indices except between EIRF and EIVL (|r|≤ 0.253, P ≥ 0.162). EIRF was negatively correlated with normalized RPD and RVD (r ≤ - 0.361, P ≤ 0.050). ICW/TW was positively correlated with normalized peak power (r = 0.421, P = 0.020). Specific muscle strength was positively correlated with absolute peak power and RPD (r ≥ 0.452, P ≤ 0.012). CONCLUSION: Knee extension power-related measures were lower in participants with higher EI, lower ICW/TW, and lower specific muscle strength, but the muscle quality indices may be determined by independent physiological characteristics.

Trade-Off Between Maximal Power Output and Fatigue Resistance of the Knee Extensors for Older Men.

This study investigated associations of fatigue resistance determined by an exercise-induced decrease in neuromuscular power with prefatigue neuromuscular strength and power of the knee extensors in 31 older men (65-88 years). A fatigue task consisted of 50 consecutive maximal effort isotonic knee extensions (resistance: 20% of prefatigue isometric maximal voluntary contraction torque) over a 70° range of motion. The average of the peak power values calculated from the 46th to 50th contractions during the fatigue task was normalized to the prefatigue peak power value, which was defined as neuromuscular fatigue resistance. Neuromuscular fatigue resistance was negatively associated with prefatigue maximal power output (r = -.530) but not with prefatigue maximal voluntary contraction torque (r = -.252). This result highlights a trade-off between prefatigue maximal power output and neuromuscular fatigue resistance, implying that an improvement in maximal power output might have a negative impact on neuromuscular fatigue resistance.

Disruption of the mitochondria-associated ER membrane (MAM) plays a central role in palmitic acid-induced insulin resistance.

The mitochondria-associated ER membrane (MAM) is a specialized subdomain of ER that physically connects with mitochondria. Although disruption of inter-organellar crosstalk via the MAM impairs cellular homeostasis, its pathological significance in insulin resistance in type 2 diabetes mellitus remains unclear. Here, we reveal the importance of reduced MAM formation in the induction of fatty acid-evoked insulin resistance in hepatocytes. Palmitic acid (PA) repressed insulin-stimulated Akt phosphorylation in HepG2 cells within 12h. Treatment with an inhibitor of the ER stress response failed to restore PA-mediated suppression of Akt activation. Mitochondrial reactive oxygen species (ROS) production did not increase in PA-treated cells. Even short-term exposure (3h) to PA reduced the calcium flux from ER to mitochondria, followed by a significant decrease in MAM contact area, suggesting that PA suppressed the functional interaction between ER and mitochondria. Forced expression of mitofusin-2, a critical component of the MAM, partially restored MAM contact area and ameliorated the PA-elicited suppression of insulin sensitivity with Ser473 phosphorylation of Akt selectively improved. These results suggest that loss of proximity between ER and mitochondria, but not perturbation of homeostasis in the two organelles individually, plays crucial roles in PA-evoked Akt inactivation in hepatic insulin resistance.

Home-based resistance training performed at either fast or slow speeds improves power output in older adults.

PURPOSE: We investigated the effect of an unsupervised, body mass- home-based resistance training program in older adults performed at either a fast or slow contractile speed on changes to muscle-power, -volume, -architecture, and fatigue resistance of the knee extensors. METHODS: Thirty-two male older adults (age 65-88 years) were separated into 1) fast-speed exercise (Fast-group), 2) slow-speed exercise (Slow-group), and 3) no exercise (Control-group) groups. Participants in the exercise groups performed 30-45 repetitions of knee-extension and sit-to-stand exercises 3 times a week for 8 weeks with different exercise speed between the groups. Before and after the intervention period, the following variables were measured: Isotonic power, isometric strength, twitch contractile properties, muscle-activity, -architecture, and -quality, neuromuscular fatigue resistance of the knee extensors, and thigh muscle volume. RESULTS: Peak power was increased in both the Fast-group (+24 %, P < 0.01, d = 0.65) and Slow-group (+12 %, P < 0.05, d = 0.33) but not in the Control-group. Training increased pennation angle of the vastus lateralis in both the Fast-group (+8 %, P < 0.01, d = 0.42) and Slow-group (+8 %, P < 0.01, d = 0.42), while only the Fast-group showed increase in pennation angle of the rectus femoris (+12 %, P < 0.01, d = 0.64) and thigh muscle volume (+16 %, P < 0.01, d = 0.52). There was no time × group interaction effect for the other neuromuscular measures. CONCLUSIONS: Unsupervised, body mass- and home-based resistance training performed at either fast or slow speeds can improve muscle power in older adults, while fast-speed exercise may be preferable over slow-speed owing to the relatively greater improvement of muscle-power, -volume, -architecture, and better time efficiency.

Can phase angle from bioelectrical impedance analysis associate with neuromuscular properties of the knee extensors?

Maintenance and improvement of neuromuscular functions is crucial for everyone regardless of age. An easy way to assess neuromuscular properties without muscle contraction is useful especially for those who cannot perform strenuous muscular force production, such as older adults and patients with orthopedic or cognitive disorders. Bioelectrical impedance analysis (BIA) can assess body electrical properties e.g., phase angle (PhA) which is regarded as muscle quantity/quality index. The purpose of this study was to investigate associations of PhA with neuromuscular properties of the knee extensors in 55 young (n = 23) and older (n = 32) adults. The values of PhA of the right thigh and whole-body were determined with BIA at 50 kHz. The participants performed 4-s maximal voluntary isometric contraction (MVIC) to measure peak torque (PTMVIC), and 1-s brief MVIC to assess rate of torque development (RTD) over the time interval of 0-200 ms. As markers of physiological mechanisms of muscle force production, twitch contractile properties (peak twitch torque, rate of twitch torque development, and time-to-peak twitch torque) of the knee extensors obtained by femoral nerve electrical stimulation, and muscle activity assessed as root mean square values of electromyographic activity (EMG-RMS) during PTMVIC and RTD measurements were measured. Thigh and whole-body PhA significantly correlated with PTMVIC (r ≥ 0.555, p < 0.001) and electrically evoked twitch parameters (peak twitch torque, rate of twitch torque development, and time-to-peak twitch torque; |r| ≥ 0.420, p ≤ 0.001), but not RTD (r ≤ 0.237, p ≥ 0.081) or EMG-RMSs (|r| ≤ 0.214, p ≥ 0.117). Stepwise multiple linear regression analysis revealed that thigh PhA was selected as a significant variable to predict PTMVIC but not RTD. Whole-body PhA was not selected as a significant variable to predict PTMVIC or RTD. In conclusion, both thigh and whole-body PhA can associate with maximal voluntary muscle strength of the knee extensors, and this association may be due to intrinsic contractile properties but not neural aspects. Regarding prediction of the knee extensor strength, thigh PhA is preferable as the predictor rather than whole-body PhA which is used as a widely acknowledged indicator of sarcopenia.

Expression of Saccharomyces cerevisiae cDNAs to enhance the growth of non-ethanol-producing S. cerevisiae strains lacking pyruvate decarboxylases.

Metabolic engineering of Saccharomyces cerevisiae often requires a restriction on the ethanol biosynthesis pathway. The non-ethanol-producing strains, however, are slow growers. In this study, a cDNA library constructed from S. cerevisiae was used to improve the slow growth of non-ethanol-producing S. cerevisiae strains lacking all pyruvate decarboxylase enzymes (Pdc-, YSM021). Among the obtained 120 constructs expressing cDNAs, 34 transformants showed a stable phenotype with quicker growth. Sequence analysis showed that the open reading frames of PDC1, DUG1 (Cys-Gly metallo-di-peptidase in the glutathione degradation pathway), and TEF1 (translational elongation factor EF-1 alpha) genes were inserted into the plasmids of 32, 1, and 1 engineered strains, respectively. DUG1 function was confirmed by the construction of YSM021 pGK416-DUG1 strain because the specific growth rate of YSM021 pGK416-DUG1 (0.032 ± 0.0005 h-1) was significantly higher than that of the control strains (0.029 ± 0.0008 h-1). This suggested that cysteine supplied from glutathione was probably used for cell growth and for construction of Fe-S clusters. The results showed that the overexpression of cDNAs is a promising approach to engineer S. cerevisiae metabolism.

Heterologous expression of bacterial phosphoenol pyruvate carboxylase and Entner-Doudoroff pathway in Saccharomyces cerevisiae for improvement of isobutanol production.

Bacterial phosphoenol pyruvate carboxylase (PPC) and enzymes in the Entner-Doudoroff (ED) pathway were heterologously expressed in Saccharomyces cerevisiae to improve the NADPH supply required for the bio-production of chemicals such as isobutanol. The heterologous expression of PPC from Synechocystis sp. PCC6803 increased in the isobutabol titer 1.45-fold (93.2±1.6 mg/L) in metabolically engineered S. cerevisiae strains producing isobutanol. This result suggested that the pyruvate and NADPH supply for isobutanol biosynthesis was activated by PPC overexpression. On the other hand, the expression of two enzymes organizing the ED pathway (6-phosphogluconate dehydratase [6PGD] and 2-dehydro-3-deoxy-phosphogluconate aldolase [KDPGA]) had no effect to isobutabol bio-production. Further analysis, however, revealed that additional expression of 6PGD and KDPGA improved the growth rate of S. cerevisiae strain BY4742 gnd1Δ. A 13C-labeling experiment using [1-13C] glucose also suggested that metabolic flow levels in the ED pathway increased slightly with the additional expression. These results showed that the ED pathway was successfully constructed in S. cerevisiae, even though activity of the pathway was too weak to improve isobutanol biosynthesis.

Protein refolding assisted by self-assembled nanogels as novel artificial molecular chaperone.

Molecular chaperone-like activity for protein refolding was investigated using nanogels of self-assembly of cholesterol-bearing pullulan. Nanogels effectively prevented protein aggregation (i.e. carbonic anhydrase and citrate synthase) during protein refolding from GdmCl denaturation. Enzyme activity recovered in high yields upon dissociation of the gel structure in which the proteins were trapped, by the addition of cyclodextrins. The nanogels assisted protein refolding in a manner similar to the mechanism of molecular chaperones, namely by catching and releasing proteins. The nanogels acted as a host for the trapping of refolded intermediate proteins. Cyclodextrin is an effector molecule that controls the binding ability of these host nanogels to proteins. The present nanogel system was also effective at the renaturation of inclusion body of a recombinant protein of the serine protease family.

Thermoresponsive controlled association of protein with a dynamic nanogel of hydrophobized polysaccharide and cyclodextrin: heat shock protein-like activity of artificial molecular chaperone.

Dynamic CHP-CD nanogels, which consisted of a self-assembly of cholesteryl-group-bearing pullulan (CHP) and beta-cyclodextrin (CD), were characterized by SEC and SEC-MALS methods. The nanogels prevented the thermal aggregation of carbonic anhydrase B (CAB) by selective trapping of the heat-denatured protein. After the complex between the CHP-CD nanogels and CAB was cooled, the enzyme activity of CAB spontaneously recovered upon release from the complex. The dynamic nanogels self-regulated an association of heat denatured protein and dissociation of native protein depending on the concentration of CD. The thermal stability of CAB was improved by thermoresponsive controlled association between the proteins and the artificial molecular chaperone.

Photoresponsive nanogels formed by the self-assembly of spiropyrane-bearing pullulan that act as artificial molecular chaperones.

Novel photoresponsive nanogels were prepared by the self-assembly of spiropyrane-bearing pullulan (SpP). The solution properties of the nanogels could be controlled by photostimulation via isomerization between hydrophobic spiropyrane and hydrophilic merocyanine. The molecular chaperone-like activity of the nanogels in protein refolding was investigated. The activity of citrate synthase significantly increased when the amphiphilicity of SpP nanogels was switched by photostimulation.