• Journal of Nutrition and Health
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• v.57 no.1
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• pp.53-64
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• 2024

Purpose: Mitochondria play a crucial role in preserving skeletal muscle mass, and damage to mitochondria leads to muscle mass loss. This study investigated the effects of oxypeucedanin hydrate, a furanocoumarin isolated from Angelica dahurica radix, on myogenesis and mitochondrial function in vitro and in zebrafish models. Methods: C2C12 myotubes cultured in media containing 0.1, 1, 10, or 100 ng/mL oxypeucedanin hydrate were immunostained with myosin heavy chain (MHC), and then multinucleated MHC-positive cells were counted. The expressions of markers related to muscle differentiation, muscle protein degradation, and mitochondrial function were determined by quantitative reverse transcription polymerase chain reaction. To investigate the effects of oxypeucedanin hydrate on mitochondrial dysfunction, Tg(Xla.Eef1a1:mito-EGFP) zebrafish embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without oxypeucedanin hydrate and analyzed for mito-EGFP intensity and mitochondrial length. Results: Oxypeucedanin hydrate significantly increased MHC-positive multinucleated myotubes (≥ 3 nuclei) and increased the expression of the myogenic marker myosin heavy chain 4. However, it decreased the expressions of muscle-specific RING finger protein 1 and muscle atrophy f-box (markers of muscle protein degradation). Furthermore, oxypeucedanin hydrate enhanced the expressions of markers of mitochondrial biogenesis (peroxisome proliferator-activated receptor-gamma coactivator 1 alpha, transcription factor a mitochondrial, succinate dehydrogenase complex flavoprotein subunit A, and cytochrome c oxidase subunit 1) and mitochondrial fusion (optic atrophy 1). However, it reduced the expression of dynamin-related protein 1 (a mitochondrial fission regulator). Consistently, oxypeucedanin hydrate reduced FOLFIRI-induced mitochondrial dysfunction in the skeletal muscles of zebrafish embryos. Conclusion: The study indicates that oxypeucedanin hydrate promotes myogenesis by improving mitochondrial function, and thus, suggests oxypeucedanin hydrate has potential use as a nutritional supplement that improves muscle mass and function.

• Journal of Nutrition and Health
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• v.56 no.6
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• pp.602-614
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• 2023

Purpose: The balance between synthesis and degradation of proteins plays a critical role in the maintenance of skeletal muscle mass. Mitochondrial dysfunction has been closely associated with skeletal muscle atrophy caused by aging, cancer, and chemotherapy. Polygalacin D is a saponin derivative isolated from Platycodon grandiflorum (Jacq.) A. DC. This study aimed to investigate the effects of polygalacin D on myoblast differentiation and muscle atrophy in association with mitochondrial function in in vitro and in zebrafish models in vivo. Methods: C2C12 myoblasts were cultured in differentiation media containing different concentrations of polygalacin D, followed by the immunostaining of the myotubes with myosin heavy chain (MHC). The mRNA expression of markers related to myogenesis, muscle atrophy, and mitochondrial function was determined by real-time quantitative reverse transcription polymerase chain reaction. Wild type AB^* zebrafish (Danio rerio) embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without polygalacin D, and immunostained to detect slow and fast types of muscle fibers. The Tg(Xla.Eef1a1:mitoEGFP) zebrafish expressing mitochondria-targeted green fluorescent protein was used to monitor mitochondrial morphology. Results: The exposure of C2C12 myotubes to 0.1 ng/mL of polygalacin D increased the formation of MHC-positive multinucleated myotubes (≥ 8 nuclei) compared with the control. Polygalacin D significantly increased the expression of MHC isoforms (Myh1, Myh2, Myh4, and Myh7) involved in myoblast differentiation while it decreased the expression of atrophic markers including muscle RING-finger protein-1 (MuRF1), mothers against decapentaplegic homolog (Smad)2, and Smad3. In addition, polygalacin D promoted peroxisome proliferator-activated receptor-gamma coactivator (Pgc1α) expression and reduced the level of mitochondrial fission regulators such as dynamin-1-like protein (Drp1) and mitochondrial fission 1 (Fis1). In a zebrafish model of FOLFIRI-induced muscle atrophy, polygalacin D improved not only mitochondrial dysfunction but also slow and fast muscle fiber atrophy. Conclusion: These results demonstrated that polygalacin D promotes myogenesis and alleviates chemotherapy-induced muscle atrophy by improving mitochondrial function. Thus, polygalacin D could be useful as nutrition support to prevent and ameliorate muscle wasting and weakness.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.73-80
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• 2004

Coenzyme Q is a quinone derivative that acts as a lipid electron carrier in the respiratory chain located at mito-chondrial inner membrane in eucaryotes or plasma membrane in procaryotes and also functions as antioxidant. A putative Neurospora crassa coq-4 gene was cloned and functionally expressed in Saccharomyces cerevisiae coq4 mutant. Complemented S. cerevisaie mutant strain was able to produce coenzyme $Q_{6}$ and showed a normal growth rate. They also showed less sensitivities to polyunsaturated fatty acids such as linoleic acid or linolenic acid. The predicted sequence of N. crassa COQ4 is consisted of 347 amino acids with a molecular mass of 39.7 kDa and showed 35% identity and 52% similarity with that of S. cerevisiae.

• Journal of Nutrition and Health
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• v.55 no.4
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• pp.462-475
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• 2022

Purpose: Allomyrina dichotoma larvae are one of the approved edible insects with nutritional value and various functional and medicinal properties. Previously we have demonstrated that the Allomyrina dichotoma larval extract (ADLE) ameliorates hepatic insulin resistance in high-fat diet (HFD)-induced diabetic mice through the activation of adenosine monophosphate-activated protein kinase (AMPK). This study investigated the effects of ADLE on insulin resistance in the skeletal muscle and explored mechanisms for enhancing the glucose uptake in palmitate (PAL)-treated C2C12 myotubes. Methods: To induce insulin resistance, the differentiated C2C12 myotubes were treated with PAL (0.5 mM) for 24 hours, and then treated with a 0.5 mg/ml concentration of ADLE, and the resultant effects were measured. The expression levels of glucose transporter-4 (GLUT4), AMPK, and the mitochondrial metabolism-related proteins were analyzed by western blotting. The mRNA expression levels of lipogenesis- related genes were determined by quantitative reverse-transcriptase PCR. Results: The exposure of C2C12 myotubes to 0.5 mg/ml of ADLE increased cell viability significantly compared to PAL-treated cells. ADLE upregulated the protein expression of GLUT4 and enhanced glucose uptake in the PAL-treated cells. ADLE increased the phosphorylated AMPK in both the PAL-treated C2C12 myotubes and HFD-treated skeletal muscle. The reduced expression levels of peroxisome-proliferator-activated receptor gamma co-activator-1 alpha (PGC1α) and uncoupling protein 3 (UCP3) due to the PAL and HFD treatment were reversed by the ADLE treatment. The citrate synthase activity was also significantly increased with the PAL and ADLE co-treatment. Moreover, the mRNA and protein expressions of fatty acid synthesis-related factors were reduced in the PAL and HFD-treated muscle cells, and this effect was significantly attenuated by the ADLE treatment. Conclusion: ADLE activates AMPK, which in turn induces mitochondrial metabolism and reduces fatty acid synthesis in C2C12 myotubes. Therefore, ADLE could be useful for preventing or treating insulin resistance of skeletal muscles in diabetes.

• KSBB Journal
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• v.24 no.3
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• pp.279-286
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• 2009

Streptomyces is well known for their ability to synthesize enormous varieties of antibiotics as secondary metabolites. Among them, S. avermitilis produces avermectins, a group of antiparasitic agents used in human and veterinary medicine. However, S. avermitilis also produces oligomycin, which is a potential toxic inhibitor of oxidative phosphorylation in mammalian cells. Therefore, we decided to disrupt oligomycin synthetase gene to prevent co-production of oligomycin in S. avermitilis. To create plasmid for disruption, the smallest gene of oligomycin synthetase gene cluster was obtained by PCR from S. avermitilis chromosome. Then, apramycin resistance gene was inserted in oligomycin synthetase gene for selection. After transformation of this plasmid, oligomycin synthetase gene (olmA5) in the chromosome was displaced with disruption cassette on the plasmid via homologous recombination. As a result of this gene replacement, we obtained mutants (olmA5::apra) that no longer makes the toxic oligomycin. And the mutants confirmed by PCR and HPLC analysis. However, showed no increasement of avermectin production in the mutant was observed.

• Development and Reproduction
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• v.3 no.1
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• pp.53-58
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• 1999

The androgenic gland secretes a hormone, androgenic gland hormone, which is believed to act on the differentiation of the primary, secondary, and behavioral sex characteristics in most malacostracan crustaceans. This report presents the ultrastructural morphology of the androgenic gland in the freshwater prawn Macrobrachium nipponense. This gland, located in the coxopodite of the last pair of walking legs, was attached to the subterminal region of the sperm duct. The gland was composed of simple cellular strands, encased by a fibrous sheath. Microvilli were situated in the fibrous sheath, especially at the edge of each cellular strand. The androgenic gland cells had the large and round nucleus and rough endoplasmic reticulum arranged either in spirals or in concentric circles throughout the cytoplasm of the cell. They also had the well-developed Golgi complex and long mitochondria with flat and transverse cristae. The Golgi complex was similar to microvesicular cluster, but usually in the shape of typical dictyosomes, These features of androgenic gland cells coincides well with the protein/peptide secretion in their function. However, despite the apparent ultastructual equipment for protein/peptide secretion, no accumulation of materials secreted were noticed in the cytoplasm. Therefore it is strongly suggested that the transient transportation of the materials into the hemocoel has occurred just after synthesis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.10
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• pp.1626-1633
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• 2004

In recent, the roles of oleic acid, most abundant fatty acid in myelin, were investigated in relation to the brain functions. This study examined the effects of diets either with desirable ratios of $\omega$6/ $\omega$9 and P/M/S (mixed oil-fed group: MO) or with defficient in $\omega$3 series fatty acids (safflower oil-fed group: SO) on the oleic acid composition in RBC and brain synaptosomal, mitochondrial & microsomal phospholipids. The desirable fatty acid composition was computer-searched with different fats and oils to meet right ratios of both $\omega$6/ $\omega$3 and P/M/S. Diets were fed 3 weeks before conception and new-born pups were fed maternal milk from the same mothers and same diets until 9 wks of age. At 3 wks of age, the compositions of oleic acid in brain subcellular fractions and red blood cells were constantly remained whatever the composition of dietary fatty acids. But at 9 wks of age, the composition of oleic acid in synaptosome and mitochondria were significantly higher in MO group than SO group. The composition of oleic acid in milk was significantly higher in MO group than SO group, but in case of SO group, that of oleic acid was increased by 48%, in comparison with dietary fatty acid compositions. -9 desaturase index (18:0\longrightarrow8:1) of brain synaptosome was significantly higher in MO group than SO group at 3 and 9 weeks of ages, but that of brain microsome was higher in SO group than MO group at 9 wks of age. Taken together, the presences of oleic acid in the diet was important to maintain brain functions. The origins of oleic acid in brain may suggests two hypotheses; first, the central nervous system has priority for the uptake of oleic acid, and second the nervous system can synthesize all the oleic acid it needs, independently of its presence in the diet.