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Ethanol Extract of Schisandra chinensis (Turcz.) Baill. Reduces AICAR-induced Muscle Atrophy in C2C12 Myotubes (마우스 C2C12 근관세포에서 AICAR로 유도된 근위축에 미치는 오미자 추출물의 영향)

  • Kang, Young-Soon;Park, Cheol;Han, Min-Ho;Hong, Su-Hyun;Hwang, Hye-Jin;Kim, Byung Woo;Kim, Cheol Min;Choi, Yung Hyun
    • Journal of Life Science
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    • v.25 no.3
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    • pp.293-298
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    • 2015
  • Muscle atrophy, known as a sarcopenia, is defined as a loss of muscle mass resulting from a reduction in the muscle fiber area or density due to a decrease in muscle protein synthesis and an increase in protein breakdown. Schisandrae fructus (SF) extract of the fruits of Schisandra chinensis (Turcz) Baillon has been used as a tonic in traditional medicine for thousands of years. Although a great deal of work has been carried out on the therapeutic potential of SF, its pharmacological mechanisms of action in muscle diseases actions remain unclear. In the present study, we investigated the inhibitory effects of SF ethanol extracts on the production of muscle atrophy factors in C2C12 myotubes stimulated with 5-aminoimidazole-4-carboxamide-ribonucleotide (AICAR), an AMP-activated kinase (AMPK) activator, and sought to determine the underlying mechanisms of action. AICAR upregulated atrophy-related ubiquitin ligase muscle RING finger-1 (MuRF-1) and stimulated the levels of the forkhead box O3a (FoxO3a) transcription factor in the C2C12 myotubes. SF supplementation effectively and concentration- dependently counteracted AICAR-induced muscle cell atrophy and reversed the increased expression of MuRF-1 and FoxO3a. Our study demonstrates that SF can reverse the muscle cell atrophy caused by AICAR through regulation of the AMPK and FoxO3a signaling pathways, followed by inhibition of MuRF-1.

The Toxicological Pathologic Study of Amanita muscaria in Sprague-Dawley Rat (Amanita muscaria 경구투여 시 Sprague-Dawley Rat에서의 독성병리 연구)

  • Kim, Jin;Kim, Hyeong-Jin;Kim, So-Jung;Kim, Byeong-Soo;Kim, Sang-Ki;Park, Byung-Kwon;Park, Young-Seok;Cho, Sung-Dae;Jung, Ji-Won;Nam, Jeong-Seok;Choi, Chang-Sun;Lee, Seung-Ho;Jung, Ji-Youn
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1152-1158
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    • 2009
  • For the toxicological pathologic study of amanita muscaria, we have investigated single and repeated dose toxicity in Sprague-Dawley (SD) rats. Single dose toxicity study was identified as catalepsy, incline and tail pinch methods (control 0 mg/kg, low 3.3 mg/kg, middle 16.5 mg/kg, high 33.0 mg/kg). Repeated dose toxicity study was carried out in blood tests, serum tests and histopathological methods. Neurotoxicity - muscle paralysis, and convulsion and loss of movement - was observed at 33.0 mg/kg group in the single dose toxicity study. Dysfunction of liver and kidney were shown in the repeated oral administration of the amanita muscaria at 3${\sim}$4 weeks. Serum chemistry results revealed a marked increase of LDH [Lactate Dehydrogenase (3181.5 IU/L; normal 230-460 IU/l)], ALT [Alanine transaminase (124.0 IU/l; normal <40 IU/l)] but the kidney was normal. Histopathological results show interstitial edema and tubular epithelial necrosis in the kidney. These results suggest that amanita muscaria has a neurotoxic effect and causes dysfunction of liver and kidney in the SD rat.

Antiobesity Effect of Citrus Peel Extract Fermented with Aspergillus oryzae (Aspergillus oryzae로 생물전환한 감귤박의 항비만 효과)

  • Jeon, Hyun Joo;Yu, Sun Nyoung;Kim, Sang Hun;Park, Seul Ki;Choi, Hyeun Deok;Kim, Kwang Yeon;Lee, Sun Yi;Chun, Sung Sik;Ahn, Soon Cheo
    • Journal of Life Science
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    • v.24 no.8
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    • pp.827-836
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    • 2014
  • There is a rising trend in obesity due to various factors, including changes in eating habits, lack of exercise, and genetic and psychological factors. Citrus peel has been reported to prevent obesity via antioxidative, antihypertensive, and LDL cholesterol-lowering effects. This study investigated the effects of citrus peel extract fermented with or without Aspergillus oryzae in a mouse model of diet-induced obesity. The animals were divided into four groups: a high-fat diet group (HFD), a normal fat diet (NFD) group, a citrus peel extract (CP) group, and a citrus peel extract fermented with A. oryzae (CPA) group. The citrus peel extract improved lipid metabolism and weight loss in the high-fat diet-induced obese mouse model. As expected, the body weight was higher in the HFD group compared with the NFD, CP, and CPA groups. However, the concentrations of total cholesterol (TG) and triglyceride (TC) in the serum and liver of the CP and CPA groups were lower than in the HFD group. There were no significant differences in the HDL cholesterol concentration among the groups. Taken together, our results suggest that extract of citrus peel biotransformed with A. oryzae had more antiobesity activity than citrus peel not transformed by A. oryzae through the fermentation of metabolites.

Parkin Interacts with the PDZ Domain of Multi-PDZ Domain Protein MUPP1 (Parkin과 Multi-PDZ Domain Protein (MUPP1) 단백질 간의 PDZ 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.24 no.8
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    • pp.820-826
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    • 2014
  • The localization to specific subcellular sites and the regulation of cell surface receptors and channels are crucial for proper functioning. Postsynaptic density-95/Disks large/Zonula occludens-1 (PDZ)-domain is involved in recognition of and interaction between various proteins, by which the localization and the regulation are mediated. Multi-PDZ domain protein 1 (MUPP1) contains 13 PDZ domains. MUPP1 serves a scaffolding function for structure proteins and signaling proteins, but the mechanism how MUPP1 is stabilized and signalized has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and Parkin. Parkin is an E3 ubiquitin ligase. Loss-of-function mutations of Parkin gene are known to cause an autosomal recessive juvenile parkinsonism. Parkin bound to the $12^{th}$ PDZ domain, but not to other PDZ domains of MUPP1. The C-terminal end of Parkin has a type II PDZ-association motif, which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, Parkin co-localized with MUPP1. When co-expressed with ubiquitin in HEK-293T cells, MUPP1 has been strongly ubiquitinated by Parkin. These findings collectively suggest that MUPP1 is a novel substrate of Parkin and its function or stability could be modulated by Parkin-mediated ubiquitination.

Effects of Dietary Flammulina velutipes Mycelium on Physico-chemical Properties and Nutritional Components of Chicken Meat (팽이버섯 균사체의 급여가 닭고기의 이화학적 특성과 영양성분에 미치는 영향)

  • Lee, Sang-Bum;Kim, Ji-Hee;Jeong, Su-Youn;Wang, Tao;Cho, Seong-Keun;Shin, Teak-Soon;Kang, Han-Seok;Kim, Youn-Chil;Kim, Seon-Ku;Lee, Hong-Gu
    • Journal of Life Science
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    • v.23 no.7
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    • pp.893-903
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    • 2013
  • The purpose of this study was to evaluate the effects of dietary Flammulina velutipes mycelium (FVM) on physico-chemical properties and nutritional components of chicken meat. Ninety-six broiler chicks (HanHyup No. 3, Korea) were divided into four groups: control (basal diet), T1 (supplemented with 1% FVM), T2 (3%), and T3 (5%). Broiler chicks in each group were slaughtered at 7 weeks of age, and their breast and thigh meat were collected. Lightness ($L^*$) and yellowness ($b^*$) in the T2 and T3 groups were higher than in the control group (p<0.05). The loss of breast meat during cooking was increased in the T1, T2, and T3 groups, and the water-holding capacity of the breast meat was also decreased in these three groups (p<0.05). The water-soluble protein solubility was lower in the T1, T2, and T3 groups than in the control group (p<0.05), but the salt-soluble protein solubility in these three groups was higher than that in the control group (p<0.05). The crude fat content of the breast meat was decreased and that of the thigh meat was increased as the level of FVM was increased (p<0.05). The unsaturated fatty acid content of the breast meat was higher in the T2 and T3 groups than in the control group (p<0.05). The free amino acid content was high in the T1, T2, and T3 groups compared to the control. In conclusion, these results showed that feeding with 3% and 5% FVM increased the L and b values in thigh meat, improved the salt-soluble protein solubility, and increased the content of unsaturated fatty acids and free amino acids.

The Effect of Extract Powder from Fresh and Black Garlic on Main Components in Serum and Organs of Streptozotocin-Induced Diabetic Rats (생마늘 및 흑마늘 추출분말이 Streptozotocin 유발 당뇨 흰쥐의 혈청 및 장기 내 주요성분에 미치는 영향)

  • Kang, Min-Jung;Lee, Soo Jung;Sung, Nak Ju;Shin, Jung-Hye
    • Journal of Life Science
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    • v.23 no.3
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    • pp.432-442
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    • 2013
  • In this study, we examined the biological activity and synergistic effects of an extract powder of 1% and 3%, each fresh (FGP) and black garlic (BGP) in streptozotocin-induced diabetic rats. Blood glucose content was significantly lower in FGP and BGP groups than control group. Glycosylated Hb was significantly higher in streptozotocin induced diabetic control group than normal group, but significantly lower in FGP and BGP groups. Total cholesterol content of the FGP and BGP groups were lower than control group, but not shown the significant difference between garlic fed groups. HDL-cholesterol concentrations of the FGP and BGP fed groups were significantly higher than control group, except of 1% BGP group. LDL and VLDL-cholesterol contents were significantly lower in 3% FGP group, and the same tendency atherogenic index and cardiac risk factor. GOT, GPT, and ${\gamma}$-GTP activity of serum were lower in FGP and BGP fed groups than control group. Glycogen contents in liver significantly higher than control group, and has not significantly difference between normal group. TBARS content was no significantly difference in the liver and serum, but in the kidney, 3% FPG and BGP fed groups were significantly lower than other experimental group. DPPH radical scavenging activity of liver has not significant difference among experimental groups, but activity was higher garlic extract powder fed groups in serum and kidney. These results indicate that dietary supplements of fresh and black garlic extract powder was contributed to lower of blood glucose, loss prevention of glycogen in liver and improve of lipid metabolism.

Changes in Physicochemical Properties of Ground Pork Meat Containing Grape Peel during Refrigerated Storage (포도과피를 첨가한 분쇄돈육의 냉장저장 중 품질변화)

  • Choi, Gang-Won;Lee, Jong-Wook
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1041-1048
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    • 2016
  • This study was carried out to investigate the effect of grape peel on the physicochemical properties of ground pork stored at 4℃ for 10 d. Four types of ground pork were evaluated: T0 without grape peel, T1 with 0.3% grape peel, T2 with 0.7% grape peel, and T3 with 1.0% grape peel. The pH increased during storage, with that of T3 the lowest (p<0.05). The L-value and a-value decreased during storage, and the a-values of T2 and T3 were significantly higher than those of T0 and T1 (p<0.05). The b-values of T0 and T1 increased with a longer storage period (p<0.05), but those of T2 and T3 were not significantly changed. The TBARS (2-thiobarbituric acid reactive substances) content increased with a longer storage period, and the TBARS content of both T2 and T3 was significantly lower than that of T0 and T1 (p<0.05). DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical scavenging activity declined with a longer storage period, and the activity of T2 and T3 was significantly higher than that of T0 and T1 (p<0.05). The VBN content of T0 and T1 also increased with a longer storage period (p<0.05), but the VBN content of T2 and T3 was not significantly changed. After storage for 4 d, the water-holding capacity declined and cooking loss and hardness increased (p<0.05), and these parameters were not significantly different among any samples. Chewiness increased with a longer storage period (p<0.05). The results suggest that the addition of grape peel to ground pork can enhance its functionality.

Incidence and Spectrum of Chromosomal Abnormalities associated with Spontaneous Abortions in Korea: 470 Products of Conception over a Period of 6 Years (2005-2010) (국내 자연유산에 의한 수태산물 핵형분석에서 관찰된 염색체 이상의 발생율과 유형: 6년(2005-2010)간 수태산물 470예 분석)

  • Han, Sung-Hee;An, Jeong-Wook;Yang, Young-Ho;Kim, Young-Jin;Cho, Han-Ik;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.8 no.1
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    • pp.44-52
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    • 2011
  • Purpose: Cytogenetic analysis of spontaneous abortions (SABs) provides valuable information to establish the causes of fetal loss, information that is essential to provide accurate reproductive and genetic counseling couples. Such analysis also provides information on the frequencies and types of chromosomal abnormalities and associated risks of recurrence. However, there have only been a few reports of chromosomal abnormalities in small samples of SABs in the Korean population. Here, we report the incidence and spectrum of chromosomal abnormalities for cases of 470 SAB in Korea. Material and Methods: Between 2005 and 2010, a total of 470 products of conception (POC) resulting from SABs were submitted to our laboratory for cytogenetic analysis from various medical sites in Korea. The incidences and types of specific chromosomal abnormalities were determined. The abnormalities were distinguished by gestational age at the time of SAB and by maternal age. Results: The frequency of chromosomal abnormalities in POCs was 54.3% (255/470), including 228 (89.3%) numerical and 27 (10.7%: 3 balanced and 24 unbalanced) structural abnormalities. Among the numerical abnormalities, trisomy was predominant (67.0%), followed by monosomy X (12.5%), polyploidy (8.2%), triple X (0.8%), and autosomal monosomy (0.8%). The overall sex ratio (male: female) among the 470 POCs with normal and abnormal karyotypes were 0.58 and 0.65, respectively. Trisomies were identified for each autosome, with the exceptions of 1, 3, and 19. Among the 171 autosomal trisomies, trisomy 16 was the most common (19.9%), followed by trisomy 22 (13.5%), trisomy 21 (12.3 %), trisomy 15 (9.9%), and trisomies 18 and 13 (5.3%). The frequency of chromosomal abnormalities decreased with gestational age and increased with maternal age, but only because of increases in trisomies and complex abnormalities. Conclusions: We have presented a large collection of cytogenetic data for SABs collected during the past 6 years and provided a database for prenatal genetic counseling of parents who have experienced SABs in Korea.

Noninvasive Prenatal Diagnosis using Cell-Free Fetal DNA in Maternal Plasma: Clinical Applications

  • Yang, Young-Ho;Han, Sung-Hee;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.8 no.1
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    • pp.1-16
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    • 2011
  • Owing to the risk of fetal loss associated with prenatal diagnostic procedures (amniocentesis, chorionic villus sampling), noninvasive prenatal diagnosis (NIPD) is ultimate goal of prenatal diagnosis. The discovery of circulating cell-free fetal DNA (cffDNA) in maternal plasma in 1997 has opened up new probabilities for NIPD by Dr. Lo et al. The last decade has seen great development in NIPD. Fetal sex and fetal RhD status determination by cffDNA analysis is already in clinical use in certain countries. For routine use, this test is limited by the amount of cell-free maternal DNA in blood sample, the lack of universal fetal markers, and appropriate reference materials. To improve the accuracy of detection of fetal specific sequences in maternal plasma, internal positive controls to confirm to presence of fetal DNA should be analyzed. We have developed strategies for noninvasive determination of fetal gender, and fetal RhD genotyping using cffDNA in maternal plasma, using real-time quantitative polymerase chain reaction (RT-PCR) including RASSF1A epigenetic fetal DNA marker (gender-independent) as internal positive controls, which is to be first successful study of this kind in Korea. In our study, accurate detection of fetal gender through gestational age, and fetal RhD genotyping in RhD-negative pregnant women was achieved. In this assay, we show that the assay is sensitive, easy, fast, and reliable. These developments improve the reliability of the applications of circulating fetal DNA when used in clinical practice to manage sex-linked disorders (e.g., hemophilia, Duchenne muscular dystrophy), congenital adrenal hyperplasia (CAH), RhD incompatibility, and the other noninvasive pregnant diagnostic tests on the coming soon. The study was the first successful case in Korea using cffDNA in maternal plasma, which has created a new avenue for clinical applications of NIPD.

The development of anti-DR4 single-chain Fv (ScFv) antibody fused to Escherichia coli alkaline phosphatase (대장균의 alkaline phosphatase가 융합된 anti-DR4 single-chain Fv (ScFv) 항체의 개발)

  • Han, Seung Hee;Kim, Jin-Kyoo
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.10-17
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    • 2016
  • Enzyme immunoassay to analyze specific binding activity of antibody to antigen uses horseradish peroxidase (HRP) or alkaline phosphatase (AP). Chemical methods are usually used for coupling of these enzymes to antibody, which is complicated and random cross-linking process. As results, it causes decreases or loss of functional activity of either antibody or enzyme. In addition, most enzyme assays use secondary antibody to detect antigen binding activity of primary antibody. Enzymes coupled to secondary antibody provide a binding signal by substrate-based color development, suggesting secondary antibody is required in enzyme immunoassay. Additional incubation time for binding of secondary antibody should also be necessary. More importantly, non-specific binding activity caused by secondary antibody should also be eliminated. In this study, we cloned AP isolated from Escherichia coli (E. coli) chromosome by PCR and fused to) hAY4 single-chain variable domain fragment (ScFv) specific to death receptor (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand (TRAIL). hAY4 ScFv-AP expressed in E. coli showed 73.8 kDa as a monomer in SDS-PAGE. However, this fusion protein shown in size-exclusion chromatography (SEC) exhibited 147.6 kDa as a dimer confirming that natural dimerization of AP by non-covalent association induced ScFv-AP dimerization. In several immunoassay such as ELISA, Western blot and immunocytochemistry, it showed antigen binding activity by color development of substrates catalyzed by AP directly fused to primary hAY4 ScFv without secondary antibody. In summary, hAY4 ScFv-AP fusion protein was successfully purified as a soluble dimeric form in E. coli and showed antigen binding activity in several immunoassays without addition of secondary antibody which sometimes causes time-consuming, expensive and non-specific false binding.