As well as the classic bean-shaped structures observed in electron micrographs, mitochondria are frequently found as long, snake-shaped tubules, and extended reticular networks whose overall morphology depends on the balance of fission and fusion of mitochondrial tubules in growing cells. [1][2][3][4] Such mitochondrial morphology, size, distribution, and copy number were demonstrated to change in living cells during cellular differentiation, development, or under pathological conditions, including liver disease, muscle dystrophy, cardiomyopathy, and cancer. [5][6][7] In differentiated cells, mitochondria are often localized to specific cytoplasmic regions rather than randomly distributed. 8) Additionally, mitochondrial morphology and distribution changed by the metabolic state of the cell or during cell growth. [9][10][11][12] Although the role of mitochondria in metabolism, ATP production, and apoptosis is more widely recognized, alterations in mitochondrial morphology and abundance are also important for cellular functions. In fact, the distribution of mitochondria in dendrites of living hippocampal neurons, which are regulated by mitochondrial fission/fusion, has been shown to be an essential and limiting factor for synapse density and plasticity. 13) Detail, however, on the mitochondrial behaviour during cellular differentiation and cell cycle progression in higher eukaryotes has little been revealed.It was shown that at S-phase, there is a substantial increase in the amount of phosphorylated histone H1 and that the phosphorylated histone H1 is diffusely distributed throughout the cell nucleus.14) On the other hand, at the end of Sphase, phosphorylation of histone H3 begins immediately following the replication of the centromeric heterochromatin.15) This phosphorylation is diffusely distributed throughout the nucleus, but, initiating at the centromers, spreads throughout the genome as the cells progress to prophase. Thus, when used in combination, staining with antibodies to phosphorylated histone H1 and H3 can be used to clearly and easily resolve mixed cell populations into each of the major stages of the cell cycle (UPSTATE, 2002 Catalog, Technical Appendix pp. 365).In the present study, we investigated mitochondrial dynamics during the cell cycle progression using synchronized HeLa cells in which the cells were transfected with mitoDsRed1 to visualize mitochondria in the cells and the major stages of the cell cycle of the observed cells were resolved by staining the cells with phosphorylated histone H1 and H3 antibodies. We found that mitochondria exist as filamentous network structures throughout the cell cycle progression, changing their morphology, distribution, and abundance. MATERIALS AND METHODS Double-Thymidine Block and Cell Cycle AnalysisHeLa cells were maintained in Dulbecco's modified Eagle's medium (DMEM; GIBCO Invitrogen Corp., Carlsbad, CA, U.S.A.) plus 10% fetal calf serum at 37°C under an atmosphere of 5% CO 2 . Cells were synchronized for the G1/S boundary by a double-thymidine block p...
of the groups. These results indicate that AGE may facilitate the turnover of aerobic glucose metabolism, attenuate oxidative stress, and promote oxygen supply based on vasodilation, suggesting that AGE ameliorates the various impairments associated with physical fatigue.
More than three thousand publications in the past have confirmed the efficacy of garlic for the prevention and treatment of a variety of diseases, acknowledging and validating its traditional uses. Garlic is also used for the treatment of fatigue, although the mechanism involved remain unclear. The anti-fatigue function of garlic may be closely related to its many favorable biological and pharmacological effects. In animal studies, garlic has been shown to promote exercise endurance. Differences in the methods of processing garlic result in differences in the intensity of its anti-fatigue effect, and the most favorable form of processing has been shown to be extraction of raw garlic followed by its natural aging for a long period in a water-ethanol mixture. In human studies, it has been confirmed that garlic produces symptomatic improvement in persons with physical fatigue, systemic fatigue due to cold, or lassitude of indefinite cause, suggesting that garlic can resolve fatigue through a variety of actions. Recently, primarily in Japan, attempts have been made to measure the intensity of fatigue objectively and quantitatively using biomarkers. Currently available data strongly suggest that garlic may be a promising anti-fatigue agent, and that further studies to elucidate its application are warranted.
Lancemaside A, a triterpenoid saponin isolated from the roots of Codonopsis lanceolata, has been reported to ameliorate the reduction of blood testosterone levels induced by immobilization stress in mice. In the present study, we investigated the metabolism and absorption of lancemaside A in mice. After oral administration of lancemaside A at 100 mg/kg body weight, the unmetabolized compound appeared rapidly in plasma (t (max) = 0.5 h). Lancemaside A has a low bioavailability (1.1%) because of its metabolism by intestinal bacteria and its poor absorption in the gastrointestinal tract. Furthermore, we identified four metabolites from the cecum of mice after oral administration of lancemaside A: codonolaside II, echinocystic acid, echinocystic acid 28-O-beta-D: -xylopyranosyl-(1 --> 4)-alpha-L: -rhamnopyranosyl-(1 --> 2)-alpha-L: -arabinopyranosyl ester, and echinocystic acid 28-O-alpha-L: -rhamnopyranosyl-(1 --> 2)-alpha-L: -arabinopyranosyl ester. Among these metabolites, codonolaside II and echinocystic acid were detected in plasma, and their t (max) values were 4 and 8 h, respectively. These findings should be helpful for understanding the mechanism of the biological effect of lancemaside A.
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