• Title/Summary/Keyword: Muscle Differentiation

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DIFFERENTIATION OF ADULT STEM CELL DERIVED FROM BUCCAL FAT PAD INTO OSTEOBLAST (협부지방에서 성체 줄기세포의 분리와 골모 세포로의 분화)

  • Pyo, Sung-Woon;Park, Jang-Woo;Lee, Il-Kyu;Kim, Chang-Hyen
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.6
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    • pp.524-529
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    • 2006
  • For the repairing of bone defect, autogenous or allogenic bone grafting remains the standard. However, these methods have numerous disadvantages including limited amount, donor site morbidity and spread of diseases. Tissue engineering technique by culturing stem cells may allow for a smart solution for this problem. Adipose tissue contains mesenchymal stem cells that can be differentiate into bone, cartilage, fat or muscle by exposing them to specific growth conditions. In this study, the authors procured the stem cell from buccal fat pad and differentiate them into osteoblast and are to examine the bone induction capacity. Buccal fat-derived cells (BFDC) were obtained from human buccal fat pad and cultured. BFDC were analyzed for presence of stem cell by immunofluorescent staining against CD-34, CD-105 and STRO-1. After BFDC were differentiated in osteogenic medium for three passages, their ability to differentiate into osteogenic pathway were checked by alkaline phosphatase (ALP) staining, Alizarin red staining and RT-PCR for osteocalcin (OC) gene expression. Immunofluorescent and biochemical assays demonstrated that BFDC might be a distinguished stem cells and mineralization was accompanied by increased activity or expression of ALP and OC. And calcium phosphate deposition was also detected in their extracelluar matrix. The current study supports the presence of stem cells within the buccal fat pad and the potential implications for human bone tissue engineering for maxillofacial reconstruction.

Study on the Detection of In vivo and In vitro Hemocytic Pathway in Grasshopper, Euprepicnemis shirakii (메뚜기 체내와 체외에서 혈구생성 경로 추적에 관한 연구)

  • Chang, Byung-Soo
    • Applied Microscopy
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    • v.33 no.2
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    • pp.131-143
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    • 2003
  • A Differentiating pathway of hemocytes in vitro and in vivo of grasshopper, Euprepocnemis shirakii was described using light and electron microscopes. In the interior of body, the stem cells of the hemopoietic organ differentiated into six types of cells respectively which are prohemoyte, plasmatocyte, granulocyte I, granulocyte II, spherulocyte and oenocytoid. The formation of these hemocytes was derived from the stem cells surrounded by a reticular cell. Hemopoietic tissue cultured in the insect media differentiated different hemocytes, but none of them underwent any mitotic division. Morphological features of the cultured cells in media were essentially the same as those of the hemocytes differentiated from the stem cells in vivo. These results were shown that each stem cell could differentiate into different types of hemocytes. It was confirmed that the stem cells possessed the pluripotent differentiation ability to directly each hemocyte, and that the once formed hemocytes in vivo and in vitro didn t undergo further transformation to other hemocytes. The maintenance of circulating hemocytes in grasshopper had been depended on the widely spreading hemopoietic organ situated in the upper surface of the dorsal alary muscle and located on the first to eighth segments.

Isolation, Molecular Phylogeny, and Tissue Distribution of Four cDNAs Encoding the Apolipoprotein Multigene Family in Barred Knifejaw, Oplegnathus fasciatus (Teleostei, Perciformes)

  • Kim, Keun-Yong;Cho, Young-Sun;Kim, Sung-Koo;Nam, Yoon-Kwon
    • Fisheries and Aquatic Sciences
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    • v.11 no.2
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    • pp.88-97
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    • 2008
  • Lipoproteins are complexes of lipids and specific apolipoproteins that are involved in lipid transport and redistribution among various tissues. In this study, we isolated full-length apolipoprotein cDNA sequences encoding apolipoprotein A-I (apoA-I), apoE, apoC-II, and apo-14 kDa in barred knifejaw, Oplegnathus fasciatus. In addition, we reconstructed phylogenetic trees and investigated mRNA tissue distributions. Alignment analyses of amino acid sequences revealed that secondary structures of the polypeptides apoA-I, apoE, and apoC-II in barred knifejaw are well conserved with their teleostean and mammalian counterparts in terms of characteristic tandem repetitive units forming amphipathic ${\alpha}$-helices. Both the sequence alignment data and cleavage sites of apo-14 kDa indicated a clear differentiation between Percomorpha and Cypriniformes. Meanwhile, the phylogenetic trees of apolipoprotein sub-families suggested that the common ancestor prior to the split of the Actinopterygii (ray-finned fishes) and Sarcopterygii (tetrapods) would have possessed the primordial protein-encoding genes. Tissue distribution of each apolipoprotein transcript determined by semi-quantitative RTPCR showed that barred knifejaw apoA-I transcripts were more or less ubiquitously expressed in the liver, intestines, brain, muscle, spleen, and kidney. The most striking difference from previous observations on barred knifejaw was the ubiquitous expression of apoE across all somatic tissues. Barred knifejaw apoC-II showed tissue-specific expression in the liver and intestines, while the liver and brain were the major sites of apo-14kDa mRNA synthesis.

Myofibrillogenesis in Embryonic Chick Pectoral Muscle Cultures: An Immunoelectron Microscopic Study (계배 배양 출근세포에 있어서의 근원섬유 형성과정 :면역 전자현미경적 연구)

  • 하재청;김한도김동수
    • The Korean Journal of Zoology
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    • v.35 no.2
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    • pp.149-160
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    • 1992
  • Using the protein A-gold complex, the mvoabrillogenesis and actin localization of cultured myoblast were invastisated. In the superstructural changes of mvogenic cell during differentiation, pectoral myoblasts contained large nucleus and numerous ribosomes but no myofibrils during the first 24 hr of cultures. Mvoblast initiated to differentiate at 3-day of culture contained the primitive myofibrillar structure. At 96 hr of culture, the mvofibrillar structure showed reletively discernable Z band but pools defined A, H and M bands. The feature of sarcomeric structure showed more defined form at cultur 5 day. In the aspect of actin localization, actin wvas diffusely detected throughout the cytoplasm of myogenic cell and nucleus during the proliferating stage. At 72 hr of culture, with the appearantc oi primitive mvofibrils, gold particles were observed in surrounding of myofibrils but still presented in overall of cytoplasm, especially in the surface and lumen of endoplasmic reticulum. With the gradual increase of culture time, local distribution of actin was readily detected within cytoplasm. In the 5-day specimen of cultures, gold particles precisely indicate the sites of actin localifation within the sarcomere. These results indicate the time of onset of myofibrill appearance and the biosynthetic and incorporation pathway of actin molecules into sarcomeric structure during myofibrillogenesis. Thus, in the present study, the first mvoabrillar structure was detected at culture 3 day, and the initiation of assembly into a typical sarcmeric structure was observed at culture 5 day. It seems, however, that the course of events on myofibrillogenesis of cultured myoblasts can be changed with great dependence of culture conditions including the number and groluth rate of mononucleated mvoblasts after seeding although the fundamental process shows identical appearances.

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Co-relation between Questionnaire for Buzhongyiqi-Tang and Electrogastrography (보중익기탕 변증설문지와 위전도의 상관성 연구)

  • Kim, Sun-Hyeok;Park, Young-Bae;Park, Young-Jae;Oh, Hwan-Sup
    • The Journal of the Society of Korean Medicine Diagnostics
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    • v.13 no.2
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    • pp.34-44
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    • 2009
  • Background : Currently, as a method of standardization of prescription, questionnaire for Buzhongyiqi-Tang[補中益氣湯] was developed, and which is a Objectives : The purpose of this thesis is to testify whether differentiation of Lao Juan Shang[勞倦傷] etiology is relative to mobility of gastric smooth muscle. Methods : The subjects(20 to 65 years old; 14 males, 46 females) were isolated from drinking alcohols for 24 hours before the experiment, and fasted for 8 hours, and measured for electrogastrography(EGG) and they filled out Questionnaire for Buzhongyiqi-Tang. Results : 1. Six factors from the factor analysis of Questionnaire for Buzhongyiqi -Tang were named and classified as Spleen-Qi deficiency syndrome factor [脾虛] (factor 1), Lung-Qi deficiency syndrome factor [肺虛] (factor 2), Working factor [習慣] (factor 3), Yin-Fire factor [陰火] (factor 4), Jung-Qi deficiency syndrome factor [中氣虛] (factor 5), and Stomach-Qi deficiency syndrome factor [胃虛] (factor 6). 2. As for the reliability of Questionnaire for Buzhongyiqi-Tang, we used Cronbach's alpha coefficient. Cronbach's alpha coefficient was 0.772 for the mean of the item-total. 3. Lung-Qi deficiency syndrome factor(factor2) had significant correlation with Bradygastria Time (r=-0.312, p<0.05). 4. Working factor(factor3) had significant correlation with Bradygastria Time (r=-0.329, p<0.05). 5. Yin-Fire factor(factor4) had significant correlation with Power Ratio (r=-0.328, p<0.05). Conclusions : It is shown that Bradygastria Time and Power Ratio tended to decrease against postprandial DP increased and postprandial frequency decreased in normal case.

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Effect of Chrysanthemum Morifolium Extracts on the Synthesis of Laminin of Madin-Darby Canine Kidney Cells (감국(甘菊)이 MDCK 세포의 Laminin 합성에 미치는 영향)

  • Na, Ho-Jeong;Jeon, So-Ra;Cha, Dong-Seok;Eun, Jae-Soon;Lim, Jong-Pil;Shin, Tae-Yong;Oh, Chan-Ho;Yang, Jae-Heon;Kim, Dae-Keun;Leem, Jae-Yoon;Chae, Byeong-Suk;Kim, Sung-Zoo;Jung, Yen-Ok;Jeong, Won-Hwan;Jeon, Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.3
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    • pp.709-713
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    • 2007
  • Basement membranes (BMs) are extracellular matrices associated with epithelia, endothelia, muscle, fat and peripheral nerve. They are involved in cell survival, migration, differentiation. BMs functions also include tissue formation and provide mechanical stability as a selective barriers. Laminins are heterotrimeric glycoproteins found in BMs and have a crucial role in cell adhesion and signalling. Madin-Darby canine kidney (MDCK) cells are the best established mammalian model for studying epithelial cell biology The cells form an epithelial monolayer, with tight junctions separating an apical surface from a basolateral membrane facing the filter support and neighboring cells. In this study, using MDCK cells, the synthesis of the BM protein such as laminin with or without methanol extract of Chrysanthemum morifolium (CM) stimulation was analyzed by immunoblotting and CM showed significant increased cell density and enhanced synthesis of laminin.

Myonectin inhibits adipogenesis in 3T3-L1 preadipocytes by regulating p38 MAPK pathway

  • Park, Tae-Jun;Park, Anna;Kim, Jaehoon;Kim, Jeong-Yoon;Han, Baek Soo;Oh, Kyoung-Jin;Lee, Eun Woo;Lee, Sang Chul;Bae, Kwang-Hee;Kim, Won Kon
    • BMB Reports
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    • v.54 no.2
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    • pp.124-129
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    • 2021
  • In current times, obesity is a major health problem closely associated with metabolic disease such as diabetes, dyslipidemia, and cardiovascular disease. The direct cause of obesity is known as an abnormal increase in fat cell size and the adipocyte pool. Hyperplasia, the increase in number of adipocytes, results from adipogenesis in which preadipocytes differentiate into mature adipocytes. Adipogenesis is regulated by local and systemic cues that alter transduction pathways and subsequent control of adipogenic transcription factors. Therefore, the regulation of adipogenesis is an important target for preventing obesity. Myonectin, a member of the CTRP family, is a type of myokine released by skeletal muscle cells. Although several studies have shown that myonectin is associated with lipid metabolism, the role of myonectin during adipogenesis is not known. Here, we demonstrate the role of myonectin during adipocyte differentiation of 3T3-L1 cells. We found that myonectin inhibits the adipogenesis of 3T3-L1 preadipocytes with a reduction in the expression of adipogenic transcription factors such as C/EBPα, β and PPARγ. Furthermore, we show that myonectin has an inhibitory effect on adipogenesis through the regulation of the p38 MAPK pathway and CHOP. These findings suggest that myonectin may be a novel therapeutic target for the prevention of obesity.

Chikungunya Virus nsP2 Impairs MDA5/RIG-I-Mediated Induction of NF-κB Promoter Activation: A Potential Target for Virus-Specific Therapeutics

  • Bae, Sojung;Lee, Jeong Yoon;Myoung, Jinjong
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1801-1809
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    • 2020
  • Chikungunya virus (CHIKV) was first identified in 1952 as a causative agent of outbreaks. CHIKV is transmitted by two mosquito species, Aedes aegypti and A. albopictus. Symptoms after CHIKV infection in human are typically fever and joint pain, but can also include headache, muscle pain, joint swelling, polyarthralgia, and rash. CHIKV is an enveloped single-stranded, positive-sense RNA virus with a diameter of approximately 70 nm. The pathogenesis of CHIKV infection and the mechanism by which the virus evades the innate immune system remain poorly understood. Moreover, little is known about the roles of CHIKV-encoded genes in the viral evasion of host immune responses, especially type I interferon (IFN) responses. Therefore, in the present study, we screened CHIKV-encoded genes for their regulatory effect on the activation of nuclear factor kappa B (NF-κB), a critical transcription factor for the optimal activation of IFN-β. Among others, non-structural protein 2 (nsP2) strongly inhibited melanoma differentiation-associated protein 5 (MDA5)-mediated induction of the NF-κB pathway in a dose-dependent manner. Elucidation of the detailed mechanisms of nsP2-mediated inhibition of the MDA5/RIG-I signaling pathway is anticipated to contribute to the development of virus-specific therapeutics against CHIKV infection.

Standardization and unification of the terms and conditions used for diagnosis in oriental medicine III (한의진단명과 진단요건의 표준화 연구III - 3차년도 연구결과 보고 -)

  • Choi, Sun-Mi;Yang, Ki-Sang;Choi, Seung-Hoon;Park, Kyung-Mo;Park, Jong-Hyun;Shim, Bum-Sang;Kim, Sung-Woo;Roh, Seok-Seon;Lee, In-Seon;Cheong, Jin-Hong;Lee, Jin-Yong;Kim, Dal-Rae;Im, Hyeong-Ho;Kim, Yun-Beom;Park, Seong-Sik;Song, Tae-Won;Kim, Jong-U;Lee, Seung-Gi;Choe, Yun-Jeong;Sin, Sun-Sik
    • Korean Journal of Oriental Medicine
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    • v.3 no.1
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    • pp.41-65
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    • 1997
  • The diagnostic requirements were suggested and explained regarding the systems of differentiation of symptoms and signs in the third year study of standardization and unification of the terms and conditions used for diagnosis in oriental medicine. The systems were as follows : - analysing and differentiating of epidemic febrile disease - analysing and differentiating in accordance with the Sasang constitutional medicine baced on four-type recognition - differentiation of diease according to pathological changes of Chong and Ren channels - standards for diagnosis of women's disease - standards for diagnosis of children's disease - standards for diagnosis of motor and sensor disturbance(-muscle born joint etc.) - standards for diagnosis of neuropsychiatric diease - standards for diagnosis of five sense organ diease - standards for diagnosis of external disease. The indivisual diagnosis pattern was arranged by the diagnostic requirments in the following odor : another name, notion of diagnosis pattern, index of differentiation of symptoms and signs, the main point of diagnosis, analysis of diagnosis pattern, discrimination of diagnosis pattrrn, prognosis, a way of curing a diseases, prescription, herbs in common use, dieases appearing the diagnosis pattern, documents. The standards for diagnosis of each disease was arranged by the diagnostic requirments in the following odor. another name, notion of diease, the main point of diagnosis, analysing and differentiating of disease, analysis of diease, discrimination of disease, prognosis, a way of curing and prescription of disease, dieases In western medicine appearing the disease in oriental medicine, documents.

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Mammalian Cloning by Nuclear transfer, Stem Cell, and Enzyme Telomerase (핵치환에 의한 cloning, stem cell, 그리고 효소 telomerase)

  • 한창열
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.423-428
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    • 2000
  • In 1997 when cloned sheep Dolly and soon after Polly were born, it had become head-line news because in the former the nucleus that gave rise to the lamb came from cells of six-year-old adult sheep and in the latter case a foreign gene was inserted into the donor nucleus to make the cloned sheep produce human protein, factor IX, in e milk. In the last few years, once the realm of science fiction, cloned mammals especially in livestock have become almost commonplace. What the press accounts often fail to convey, however, is that behind every success lie hundreds of failures. Many of the nuclear-transferred egg cells fail to undergo normal cell divisions. Even when an embryo does successfully implant in the womb, pregnancy often ends in miscarriage. A significant fraction of the animals that are born die shortly after birth and some of those that survived have serious developmental abnormalities. Efficiency remains at less than one % out of some hundred attempts to clone an animal. These facts show that something is fundamentally wrong and enormous hurdles must be overcome before cloning becomes practical. Cloning researchers now tent to put aside their effort to create live animals in order to probe the fundamental questions on cell biology including stem cells, the questions of whether the hereditary material in the nucleus of each cell remains intact throughout development, and how transferred nucleus is reprogrammed exactly like the zygotic nucleus. Stem cells are defined as those cells which can divide to produce a daughter cell like themselves (self-renewal) as well as a daughter cell that will give rise to specific differentiated cells (cell-differentiation). Multicellular organisms are formed from a single totipotent stem cell commonly called fertilized egg or zygote. As this cell and its progeny undergo cell divisions the potency of the stem cells in each tissue and organ become gradually restricted in the order of totipotent, pluripotent, and multipotent. The differentiation potential of multipotent stem cells in each tissue has been thought to be limited to cell lineages present in the organ from which they were derived. Recent studies, however, revealed that multipotent stem cells derived from adult tissues have much wider differentiation potential than was previously thought. These cells can differentiate into developmentally unrelated cell types, such as nerve stem cell into blood cells or muscle stem cell into brain cells. Neural stem cells isolated from the adult forebrain were recently shown to be capable of repopulating the hematopoietic system and produce blood cells in irradiated condition. In plants although the term$\boxDr$ stem cell$\boxUl$is not used, some cells in the second layer of tunica at the apical meristem of shoot, some nucellar cells surrounding the embryo sac, and initial cells of adventive buds are considered to be equivalent to the totipotent stem cells of mammals. The telomere ends of linear eukaryotic chromosomes cannot be replicated because the RNA primer at the end of a completed lagging strand cannot be replaced with DNA, causing 5' end gap. A chromosome would be shortened by the length of RNA primer with every cycle of DNA replication and cell division. Essential genes located near the ends of chromosomes would inevitably be deleted by end-shortening, thereby killing the descendants of the original cells. Telomeric DNA has an unusual sequence consisting of up to 1,000 or more tandem repeat of a simple sequence. For example, chromosome of mammal including human has the repeating telomeric sequence of TTAGGG and that of higher plant is TTTAGGG. This non-genic tandem repeat prevents the death of cell despite the continued shortening of chromosome length. In contrast with the somatic cells germ line cells have the mechanism to fill-up the 5' end gap of telomere, thus maintaining the original length of chromosome. Cem line cells exhibit active enzyme telomerase which functions to maintain the stable length of telomere. Some of the cloned animals are reported prematurely getting old. It has to be ascertained whether the multipotent stem cells in the tissues of adult mammals have the original telomeres or shortened telomeres.

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