• International Journal of Stem Cells
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• v.16 no.1
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• pp.78-92
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• 2023

Background and Objectives: This study aims to clarify the systems underlying regulation and regulatory roles of hydrogen combined with 5-Aza in the myogenic differentiation of adipose mesenchymal stem cells (ADSCs). Methods and Results: In this study, ADSCs acted as an in vitro myogenic differentiating mode. First, the Alamar blue Staining and mitochondrial tracer technique were used to verify whether hydrogen combined with 5-Aza could promote cell proliferation. In addition, this study assessed myogenic differentiating markers (e.g., Myogenin, Mhc and Myod protein expressions) based on the Western blotting assay, analysis on cellular morphological characteristics (e.g., Myotube number, length, diameter and maturation index), RT-PCR (Myod, Myogenin and Mhc mRNA expression) and Immunofluorescence analysis (Desmin, Myosin and 𝛽-actin protein expression). Finally, to verify the mechanism of myogenic differentiation of hydrogen-bound 5-Aza, we performed bioinformatics analysis and Western blot to detect the expression of p-P38 protein. Hydrogen combined with 5-Aza significantly enhanced the proliferation and myogenic differentiation of ADSCs in vitro by increasing the number of single-cell mitochondria and upregulating the expression of myogenic biomarkers such as Myod, Mhc and myotube formation. The expressions of p-P38 was up-regulated by hydrogen combined with 5-Aza. The differentiating ability was suppressed when the cells were cultivated in combination with SB203580 (p38 MAPK signal pathway inhibitor). Conclusions: Hydrogen alleviates the cytotoxicity of 5-Aza and synergistically promotes the myogenic differentiation capacity of adipose stem cells via the p38 MAPK pathway. Thus, the mentioned results present insights into myogenic differentiation and are likely to generate one potential alternative strategy for skeletal muscle related diseases.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.2
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• pp.114-120
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• 1994

To clarify the effect of storage temperature on the morphological and histological changes of plaice, Paralichthys olivaceus muscle at early stages after killing, the changes in breaking strength of muscle, morphological observation of myofibrils and histological observation of extracellular spaces during storage at $0^{\circ}C\;and\;10^{\circ}C$ were studied. The maximum breaking strength of samples stored at $0^{\circ}C$ was reached within 10hrs and then it dropped significantly (p<0.05) from 10hrs to 25hrs of storage. However, breaking strength was not increased in fresh muscle stored at $10^{\circ}C$ and gradually decreased after 10hrs storage. In myofibrils prepared from dorsal muscle immediately after death, A-band, I-band, H-band and Z-line in sarcomere were clearly distinguishable from each other. Due to muscle contraction, it was not easy to distinguish H-band from I-band observed in sarcomere stored at $0^{\circ}C$ after 10hrs storage. But, in the case of samples stored at $10^{\circ}C$, H-band could be observed dimly until 15hrs of storage. The changes in morphological myofibrils were closely related to increase of breaking strength. No extracellular space was observed among muscle cells immediately after killing. Stored samples at $0^{\circ}C$ showed extracellular spaces after 15hrs storage. On the other hand, samples stored at $10^{\circ}C$ didn't show any extracellular spaces until 15hrs storage and showed extracellular spaces after 24hrs storage. It was thought that the post-mortem tenderization of plaice muscle was closely related to the gradually disintegration of the extracellular matrix structure after killing.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.4
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• pp.327-334
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• 1994

This study was undertaken to clarify the effect of killing methods on the morphological and histological changes of plaice, Paralichthys olivaceus muscle at early stage after killing. Killed samples by the three different methods were stored at $5^{\circ}$, and the changes in breaking strength of muscle, morphological observation of myofibrils and histological observation of extracellular spaces through storage were monitored. Samples killed by electrifying in sea water showed the maximum value of breakin strength immediately after killing and then it dropped significantly(p<0.05) until 2.5hrs passed. Breaking strength of samples killed by spiking at the head instantly and dipping in sea water including anesthetic rose steadily over 10hrs and 15hrs after killing, respectively. In myofibrills prepared from dorsal muscles immediately after spiking at the head instantly, A-band, H-band, I-band, and Z-line in sarcomere were clearly distinguishable each other. Due to muscle contraction by electrical stimulation, it was impossible to distinguish H-band from I-band observed in sarcomere immediately after killing for samples killed by electrifying. But, in the cases of samples killed by spiking and dipping, H-band could be observed dimly until 10hrs and 15hrs storage. No extracellular space was observed among muscle cells immediately after spiking at the head instantly. Samples killed by spiking at the head instantly and dipping in sea water including anesthetic showed extracellular spaces among all muscle cells after 15hrs and 25hrs storage, respectively. The other hand, samples killed by electrifying in sea water (110V, 30sec.) showed a few extracellular spaces immediately after killing and then it showed extracellular spaces among all muscle cells after 2.5hrs storage.

• Korean Journal of Ichthyology
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• v.23 no.1
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• pp.1-9
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• 2011

Differentially Expressed Gene (DEG) was obtained from Differential Display Reverse Transcription (DDRT)-PCR using Annealing Control Primer (ACP) to search and clone genes related to developmental stages of Sebastes inermis. By using 120 ACPs, the nucleotide sequences obtained from 16 DEGs showing higher expression in 6-month-old skeletal muscle than 18-month-old ones and from 22 DEGs displaying stronger expression in 18-month-old than 6-month-old were analyzed and BLAST was conducted. The results identified that DEGs shared 69~95% homology with genes of parvalbumin (PVALB), nucleoside diphosphate kinase (NDK) B, tropomyosin (TPM), troponin I (TnI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), muscle-type creatine kinase (CKM2), small EDRK-rich factor 2 (SERF2), adenosine monophosphate deaminase (AMPD), Trimeric intracellular cation channel type A (TRICA), Rho GTPase-activating protein 15 (ARHGAP15), S-formylglutathione hydrolase (Esterase D; ESD), heat shock protein 70 (hsp70), type 1 collagen alpha 2 (COL1A2), glutathione S-transferase, Mid1-interacting protein 1 (Mid1lip1), myosin light chain 1 (MYL1), sarcoplasmic/endoplasmic reticulum calcium ATPase 1B (SERCA1B), and ferritin heavy subunit (FTH1). Expression pattern by developmental stage of DEG14 and PVALB exhibiting strong expression in 6-month-old skeletal muscle was investigated using real time PCR. Expression was reduced as Sebastes inermis grew. Expression of PVALB gene was extremely low after 6 months of age. Expression of CKM2 showed higher expression in 18-month-old skeletal muscle than in 6-month-old muscles, and increased continuously until 4 years old, after which CKM2 expression became gradually reduced. By analysis of tissue-specific expression patterns of DEG, DEG14 was expressed mainly in skeletal muscle, liver, kidney and spleen tissues, whereas PVALB expression was expressed in skeletal muscle and kidney, but not in liver and spleen tissues. CKM2 was expressed in skeletal muscle, kidney, and spleen tissues, but not in liver tissues. PVALB gene was composed of 110 amino acids, which constituted 659 bp nucleotides. The results reported here demonstrate that the expression patterns of parvalbumin and CKM2 could be used as molecular markers for selecting fishes exhibiting fast growth.

• Development and Reproduction
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• v.11 no.3
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• pp.167-177
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• 2007

In the present study, we isolated three human adult stem cells including adipose tissue-derived stem cells(HAD), umbilical cord-derived stem cells(HUC), and amnion-derived stem cells(HAM) and analysed their characteristics. And we examined whether HAD could be used as therapeutical cells for the heart diseases. Both HAM and HUC appeared very similar morphology but HAD was different. Doubling time of HUC was most fast, but total doubling numbers of HUC was same with HAM. Total doubling numbers of HAD was much more than others. Expression patterns of genes and proteins of three human adult stem cells were very similar. Also they were differentiated into adipocytes, osteocytes, and chondrocytes. In addition, they expressed many cardiomyocyte-related genes. But expression pattern of genes is a little different. When HAD were cultivated in the presence or absence of various combinations of BMP and FGF after 5-azacytidine expose for 24 h, expression of Cmlc-1, and ${\alpha}1c$ genes was significantly increased. However, expression of troponin T, troponin I and Kv4.3 genes was not changed. Based on these observations, it is suggested that HAD, HUC, and HAM might be used as potentially therapeutical cells for clinical application.

• Food Science and Preservation
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• v.14 no.3
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• pp.288-293
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• 2007

Intermediate moisture-apple snail products were prepared by adding glycerol, sorbitol, kiwi, or pineapple(2% or 5% w/w), as tenderizers, and by drying at $4^{\circ}C$ for 24 hr. The effects of the tenderizers on textural and sensory properties of the apple snail products at intermediate moisture levels were investigated. Moisture content and water activity of the products were ranged from 26.25 to 34.48% and from 0.83 to 0.87, respectively. The addition of glycerol significantly lowered water activity of apple snail samples compared to control prepared without tenderizers. On the other hand, significant increases in moisture content and water activity were observed in apple snail samples treated with kiwi or pineapple(p<0.05). All apple snail samples treated with tenderizers showed a lower shear force than did the control. Apple snail samples treated with 5%(v/v) glycerol showed a higher equilibrium moisture content than did the other samples. SDS-PAGE indicated that proteolytic enzymes in kiwi and pineapple clearly changed the structure of the myosin heavy chain and actin filaments of myofibrillar protein in apple snail samples. Intermediate moisture apple snail samples treated with tenderizers showed significantly improved overall sensory characteristics. The highest overall acceptability was obtained from apple snail samples treated with 5% pineapple, while the lowest overall acceptability was noted in the control sample. This study demonstrates that an acceptable apple snail, with intermediate moisture content, may be produced by using tenderizers at appropriate concentrations.

• Journal of Life Science
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• v.31 no.7
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• pp.626-630
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• 2021

This study was conducted to examine the association between the myosin heavy chain 3 (MYH3) in 6-bp deletion variant genotypes and meat color traits in a crossbred pig population Landrace and Jeju native black pigs (JNBP). The longissimus dorsi, semimembranosus, triceps brachii and biceps femoris muscle from each carcass were used for the analysis of meat color traits. A total of 187 pigs and three meat color traits, CIE L^* (lightness), CIE a^* (redness), and CIE b^* (yellowness), were analyzed. All experimental pigs were successfully genotyped for the MYH3 6-bp deletion variant using Polymerase chain reaction (PCR) analysis. We detected three MYH3 6-bp deletion variant genotypes qq, Qq, and QQ with 0.091, 0.551 and 0.358 genotype frequencies, respectively. Compared to qq homozygotes, the MYH3 6-bp deletion QQ genotype animals showed a higher levels of the meat colors traits CIE L^* (lightness), CIE a^* (redness), and CIE b^* (yellowness) in longissimus dorsi (p>0.05, p<0.001, p<0.001), semimembranosus (p>0.05, p<0.001, p<0.001), triceps brachii (p<0.001, p<0.001, p<0.001), and biceps femoris (p<0.01, p<0.001, p<0.001), respectively. The QQ genotype pigs was associated with increasing meat color traits in the crossbred between Landrace and JNBP. Our findings suggest that the MYH3 6-bp deletion variant genotypes can be used as valuable genetic markers for JNBP-related breeding programs to improve meat quality and control meat color traits.

• 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.