• Title/Summary/Keyword: C2C12 differentiation

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EFFECT OF HEPATOCYTE GROWTH FACTOR ON THE REPAIR OF DEFECT IN THE ARTICULAR DISC IN RABBIT TEMPOROMANDIBULAR JOINT (가토의 측두하악관절원판 결손에서 간세포 성장인자가 치유에 미치는 영향)

  • Kim, Bok-Joo;Seong, Hwa-Sik;Kim, Chul-Hoon;Kim, Gyoo-Cheon;Hwang, Hee-Sung;Shin, Sang-Hun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.1
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    • pp.1-7
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    • 2009
  • Purpose: The purpose of this study is to investigate the therapeutic use of Hepatocyte growth factor(Adv.CMV.HGF) in temporomandibular joint disc defect. Materials and methods: Twelve New Zealand white rabbits, weighing 2.5 - 3.0 kg, were used in this experiment. Defects(2 mm in diameter) were created in their TMJ discs. Recombinant Adv.CMV.HGF with gelatin sponge($Gelfoam^{(R)}$) as carrier was implanted in the defects. We divided the rabbits into four batches according to the duration of the implantation - of 1, 4, 8, 12 weeks - and both left and right TMJ of each rabbit in all groups were used in the research : left joints were used as experiment group and right were control group. Each batch of rabbits was killed one, four, eight and twelve weeks after the experimentation respectively, and called Group A, B, C, and D. (Group A = 1 wk, B = 4 wks, C = 8 wks, and D = 12 wks) Results: The experimental group showed a significant increase in the number of chondroblasts and active cell differentiation at the margin of the defects. Compared to the control group, in the experiment group chondroblasts increased and chondrocytes showed a columnar arrangement, which is witnessed at the time of cell differentiation. Conclusion: This study supports the case that Avd.CMV.HGF may be useful in the repair of articular disc of the rabbit TMJ.

Culture Characteristics of Streptomyces spp. on Improved Polyacrylamide Gel and Agar Media

  • Han, Hong ui;Baek, Ji-Ho;Yang, Moon
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.384-386
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    • 1996
  • Application of polyacrylamide gel (PAG) instead of agar to solid cultures of Streptomyces spp. was studied. The improved media were prepared by 1) gelling 20 ml of 5% acrylamide in a glass petri dish at room temperature, 2) washing by running water for more than 8 hr to remove residual reaction reagents, 3) drying at 5$0^{\circ}C$ for 12 hr to make a gel film, 4) autoclaving at 121$^{\circ}C$ for 15 min, and 5) swelling gel for about 4 hr by adding sterile liquid medium. In PAG media there were no differences from the observation of morphological characteristics showing during the cellular differentiation on agar media, whereas the ability to utilize carbohydrates differed somewhat from agar media. Agar media thus were little favorable for biochemical tests which the growth was determined depending on the formation of colony, but washed PAG was superior to serve as a solidifying agent.

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The expression analysis of mouse interleukin-6 splice variants argued against their biological relevance

  • Annibalini, Giosue;Guescini, Michele;Agostini, Deborah;De Matteis, Rita;Sestili, Piero;Tibollo, Pasquale;Mantuano, Michela;Martinelli, Chiara;Stocchi, Vilberto
    • BMB Reports
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    • v.45 no.1
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    • pp.32-37
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    • 2012
  • Alternative splicing generates several interleukin-6 (IL-6) isoforms; for them an antagonistic activity to the wild-type IL-6 has been proposed. In this study we quantified the relative abundance of IL-6 mRNA isoforms in a panel of mouse tissues and in C2C12 cells during myoblast differentiation or after treatment with the $Ca^{2+}$ ionophore A23187, the AMP-mimetic AICAR and TNF-${\alpha}$. The two mouse IL-6 isoforms identified, IL-6${\delta}$5 (deletion of the first 58 bp of exon 5) and IL-6${\delta}$3 (lacking exon 3), were not conserved in rat and human, did not exhibit tissue specific regulation, were expressed at low levels and their abundance closely correlated to that of full-length IL-6. Species-specific features of the IL-6 sequence, such as the presence of competitive 3' acceptor site in exon 5 and insertion of retrotransposable elements in intron 3, could explain the production of IL-6${\delta}$5 and IL-6${\delta}$3. Our results argued against biological significance for mouse IL-6 isoforms.

Tricho-dento-osseous Syndrome Mutant Dlx3 Shows Lower Transactivation Potential but Has Longer Half-life than Wild-type Dlx3

  • Cha, Ji-Hun;Ryoo, Hyun-Mo;Woo, Kyung-Mi;Kim, Gwan-Shik;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.32 no.4
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    • pp.119-125
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    • 2007
  • Dlx3 is a homeodomain protein and is known to play a role in development and differentiation of many tissues. Deletion of four base pairs in DLX3 (NT3198) is causally related to tricho-dento-osseous (TDO) syndrome (OMIM #190320), a genetic disorder manifested by taurodontism, hair abnormalities, and increased bone density in the cranium. The molecular mechanisms that explain the phenotypic characteristics of TDO syndrome have not been clearly determined. In this study, we examined phenotypic characteristics of wild type DLX3(wtDlx3) and 4-BP DEL DLX3 (TDO mtDlx3) in C2C12 cells. To investigate how wtDlx3 and TDO mtDlx3 differentially regulate osteoblastic differentiation, reporter assays were performed by using luciferase reporters containing the promoters of alkaline phosphatase, bone sialoprotein or osteocalcin. Both wtDlx3 and TDO mtDlx3 enhanced significantly all the reporter activities but the effect of mtDlx3 was much weaker than that of wtDlx3. In spite of these differences in reporter activity, electrophoretic mobility shift assay showed that both wtDlx3 and TDO mtDlx3 formed similar amounts of DNA binding complexes with Dlx3 binding consensus sequence or with ALP promoter oligonucleotide bearing the Dlx3 binding core sequence. TDO mtDlx3 exhibits a longer half-life than wtDlx3 and it corresponds to PESTfind analysis result showing that potential PEST sequence was missed in carboxy terminal of TDO mtDlx3. In addition, co-immunoprecipitation demonstrated that TDO mtDlx3 binds to Msx2 more strongly than wtDlx3. Taken together, though TDO mtDlx3 acted as a weaker transcriptional activator than wtDlx3 in osteoblastic cells, there is possibility that during in vivo osteoblast differentiation TDO mtDlx3 may antagonize transcriptional repressor activity of Msx2 more effectively and for longer period than wtDlx3, resulting in enhancement of osteoblast differentiation.

C-reactive protein and N-terminal pro-brain natriuretic peptide discrepancy: a differentiation of adenoviral pharyngoconjunctival fever from Kawasaki disease

  • Choi, Jung Eun;Kang, Hee Won;Hong, Young Mi;Sohn, Sejung
    • Clinical and Experimental Pediatrics
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    • v.61 no.1
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    • pp.12-16
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    • 2018
  • Purpose: To differentiate adenoviral pharyngoconjunctival fever (PCF) from acute Kawasaki disease (KD) using laboratory tests before results of virus-real time polymerase chain reaction and ophthalmologic examination are obtained. Methods: Baseline patient characteristics and laboratory measurements were compared between 40 patients with adenovirus infection and 123 patients with KD. Results: The patients with adenovirus infection were generally older than those with KD (median: 3.9 years vs. 2 years, P=0.000). White blood cell and, platelet count, and aspartate aminotransferase, alanine aminotransferase, and N-terminal pro-brain natriuretic peptide (NT-proBNP) levels showed significant differences between the 2 groups, but the C-reactive protein (CRP) levels did not ($6.8{\pm}3.0mg/dL$ vs. $8.3{\pm}5.8mg/dL$, P=0.126). In the adenovirus infection group, the CRP levels were <1, <3, <10, and ${\geq}10mg/dL$ in 2 (5%), 3 (7.5%), 30 (75%), and 5 patients (12.5%), respectively. The cutoff NT-proBNP level was 265 pg/mL. Discrepancy was defined as CRP and NT-proBNP levels of ${\geq}3$ or <3 mg/dL, and <265 or ${\geq}265pg/mL$, respectively. Among the 35 patients with adenovirus infection whose CRP levels were ${\geq}3mg/dL$, 29 (82.9%) showed a discrepancy. Conversely, of the 103 patients with KD whose CRP levels were ${\geq}3mg/dL$, 83 (80.6%) showed no discrepancy. Between the groups, a significant difference in discrepancy rate was observed (P=0.000). None of the patients with adenovirus infection had CRP and NT-proBNP levels of <3 mg/dL and ${\geq}265pg/mL$, respectively. Conclusion: With a sensitivity of 82.9% and a specificity of 80.6%, CRP and NT-proBNP levels may differentiate between adenoviral PCF and acute KD.

Effects of polygalacin D extracted from Platycodon grandiflorum on myoblast differentiation and muscle atrophy (길경에서 추출한 polygalacin D가 근원세포 분화 및 근위축에 미치는 영향)

  • Eun-Ju Song;Ji-Won Heo;Jee Hee Jang;Eonmi Kim;Yun Hee Jeong;Min Jung Kim;Sung-Eun Kim
    • 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.

Characterization and In Vitro Differentiation of Korean Ring-Necked Pheasant (Phasianus colchicus) Male Germ Cells

  • Jeong, Dong Kee;Sharma, Neelesh;Nguyen, Thanh Luan;Kim, Jong Hyun;Oh, Sung Jong
    • Journal of Embryo Transfer
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    • v.29 no.4
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    • pp.351-359
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    • 2014
  • Phasianus colchicus is not only a beautiful bird but also a great value in science and under the threat of endanger. Hence, the aim of this study was to isolate the pheasant male germ cells (mGCs) and then induce them into elongated sperm-like cells in vitro. The mGCs were purified and enriched by a two-step plating method based on the different adherence velocities of mGCs and somatic cells. The percentage of the c-kit positive cells and c-kit negative cells examined by flow cytometry analysis (FCA) was 92.87% and 2.57%, respectively. Subsequently, the mGCs were induced for 48h in DMEM/F12 medium supplemented factors such as retinol acid, testosterone and bovine FSH, followed by 5 weeks in culture. We found that some elongated sperm-like cells appeared initially in vitro under inducement of stimulated factors. The elongated sperm-like cells showed in the expression of changed morphology and post-transcriptional marker such as spermatid associated (SPERT), spermatid perinuclear RNA binding protein (STRBP), round spermatid basic protein 1 (RSBN1) and SPER1L. Moreover, in DNA content identified assay, induced cells showed that the 1C DNA population markedly increased in differentiated group but it was not change in undifferentiated group. Successful in vitro differentiation of pheasant testicular germline cells into spermatids appears to offer extremely attractive potential for the conservation of endangered birds and treatment of male infertility.

Petrogenesis and Metamorphism of Charnockite of Eastern Jirisan Area (지리산 동부 지역에 분포하는 차노카이트의 변성작용과 성인에 관한 연구)

  • 김동연;송용선;박계헌
    • The Journal of the Petrological Society of Korea
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    • v.11 no.3_4
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    • pp.138-156
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    • 2002
  • Precambrian metamorphic rocks of southwest Sobaeksan massif consist of mainly granitic gneiss, porphyroblastic gneiss and quartzofeldspathic gneiss. The orthopyroxene-bearing rocks(charnockites) are found in the west of Hadong-Sancheong anorthosite complex. The charnockites are 3km wide, 12km long and divided into massive and foliated types based on their texture. The compositions of charnockites are comparable to granodiorite to adamellite and subalkaline. Variations in major and trace elemental abundances show typical magmatic differentiation trends. The geochemical data plotted on tectonic discrimination diagrams reveal that these charnockites were formed in the active tectonic environment. The massive and folidated charnockites are mainly composed of plagioclase, orthopyroxene, microcline, quartz and disseminated garnet. Camels generally show characteristic zonal textures with decreasing $X_{alm}$(0.74~0.83), $X_{Py}$ (0.07~0.12) and $X_{Mg}$ (0.12~0.08) and increasing $X_{grs}$(0.03~0.15) from core to rim. Metamorphic temperature and pressure of the charnockites estimated from orthopyroxene-garnet-plagioclase-quartz assemblages show wide range of variation of $600~900^{\circ}C$ and 2.5~7.5 kbar respectively. The results of P-T estimates indicate an anticlockwise P-T evolution path.

Expressional Evaluation of C/EBP Family, SREBP1, and Steroid Hormone Receptors in the Epididiymal Fat of Postnatally Developing Mouse

  • Lee, Yong-Seung;Lee, Ki-Ho
    • Development and Reproduction
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    • v.26 no.2
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    • pp.49-58
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    • 2022
  • The differentiation and development of preadipocyte into mature adipocyte are regulated by transcription factors, such as CCAAT enhancer binding protein (Cebp) gene family and sterol regulatory element binding transcription factor 1 (Srebp1). Steroid hormones give influences on the development and function of adipocyte. The present research examined expression patterns of CCAAT enhancer binding protein alpha (Cebpa), CCAAT enhancer binding protein beta (Cebpb), CCAAT enhancer binding protein gamma (Cebpg), sterol regulatory element binding transcription factor 1 (Srebp1), androgen receptor (Ar), and estrogen receptors (Esr) among different epididymal fat parts during postnatal period by quantitative real-time polymerase chain reaction. In the distal epididymal fat, expression of Cebpa, Cebpb, Cebpg, Srebp1, Ar, and Esr2 was increased until 12 months of age, while expression of Esr1 was decreased at 5 months of age and was not detectable after 8 months of age. In the proximal epididymal fat, transcript levels of Cebps and Srebp1 were increased at 8 months of age, followed by decreases of Cebpb and Cebpg transcript levels at 12 months of age. An additional increase of Srebp1 expression was observed at 12 months of age. Expression of Ar and Esr2 were increased until 8 months of age, followed by a drop of Ar expression level at 12 months of age. Expression pattern of Esr1 was similar to that in the distal epididymal fat. In the tail epididymal fat, expression of Cebpa, Cebpg, Srebp1, Ar, and Esr2 was increased with age. Esr1 was not detectable at all. The highest level of Cebpb was observed at 8 months of age. These data suggest the possibility of developmental and functional differentiation among the epididymal fat parts.

Id3 mRNA Expression on Folliculogenesis in Rat Ovary (쥐 난소에서 난포 발달에 따른 Id3 mRNA의 발현)

  • Hwang, Seong-Soo;Ko, Yeoung-Gyu;Lim, Hyun-Joo;Seong, Hwan-Hoo;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.181-186
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    • 2007
  • Inhibitor of DNA binding protein or inhibitor of differentiation(Id) is largely considered as positive and/or negative regulators of proliferation, differentiation, angiogeneisis, and apoptosis. The four Id genes(Id1, Id2, Id3, and Id4) were known in mammals. However, little is known about the expression and function of these genes in reproductive physiology. Among them, this study was conducted to analyze the expression pattern of Id3 mRNA on folliculogenesis in rat ovary. After PMSG administration, the ovaries were obtained at 3, 6, 12, 24, 36, and 48hrs, fixed, dehydrated, and paraffin embedded. For in situ hybridization, anti-sense and sense Id3 cRNA probes were prepared and applied to the ovarian section. The ovarian sections were coated with NTB-2 emulsion. After that, the slides were developed and counterstained with hematoxylin and eosin staining. The hybridization signal was estimated on a scale of 1+ to 4+. In oocyte, the intensity of Id3 mRNA in primordial and primary follicles was scored at ${\geq}2+$, but the intensity was less than 1+ in secondary, dominant, and preovulatory follicles. In granulosa cells, the Id3 mRNA was strongly expressed(3+ or 4+) in dominant and preovulatory follicles. Taken together, Id3 mRNA was expressed specifically at follicle stages and follicular tissue and might be closely related with follicle development.