• Title/Summary/Keyword: Protein stain

검색결과 154건 처리시간 0.021초

연쇄구균의 세포벽 단백질이 L929 세포의 미세구조 변화에 미치는 영향에 관한 연구 (THE EFFECT OF CELL WALL PROTEINS OF STREPTOCOCCUS SPECIES ON MICROSTRUCTURAL CHANGES OF L929 CELLS)

  • 오세홍;임미경
    • Restorative Dentistry and Endodontics
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    • 제20권2호
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    • pp.549-576
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    • 1995
  • Bacteria are one of the most important causative agents of the pulpal and periapical diseases. Streptococci are one of the most frequently isolated facultative anarerobic bacteria in the infected root canals. Bacterial cell wall components have a direct effect in the pathogenesis of the pulpal and periapical infections. Hyaluronidase produced by bacteria has been implicated in dissemination of the diseases. The purpose of this study was to evaluate the effect of cell wall extract of streptococci on the L929 cells using inverted microscope and the transmission electron microscopy (TEM). Hyaluronidase production of streptococcal strains were investigated to determine the correlation between the severity of cell damage and the activity of enzymes. Bacterial cell wall extracts of S. sanguis, S. mitis and S. uberis isolated from infected root canals and ATCC type strains of S. mutans (ATCC 10449) and E. faecalis (ATCC 19433) were prepared by sonication and confirmed with SDS-PAGE. Silver stain of SDS-PAGE of sonic extract was efficient at $100{\mu}g$/ml concentration of cell wall protein, while Coomasie blue stain was efficient at $100{\mu}g$/ml concentration. Inverted microscope showed that sonic extract-treated L929 cells were round and detached from the substratum while others lost their fibroblastic shapes. Transmission electron microscopic examination revealed that streptococcal extracts induced death of L929 cells. Sonic extracts of streptococci had variable effect on microstructure of L929 cells. significant chromatin condensation was observed in the nucleus of the cells. Disappearance of cell surface microvilli and nuclear fragments with dense chromatin were observed. The cell nucleus had an irregular shape and numerous large vacuoles were seen in the cytoplasm and some breaks of the cell membrane could be seen. Cell organelles were in various stages of destruction and cristae of mitochondria were disoriented or disappeared. Eighteen strains of streptococci did not produce hyaluronidase.

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분리균 Erwinia carotovora subsp. carotovora LY34의 병원성 및 CMCase Isozymes 생성

  • 임선택;박용우;조수정;윤한대
    • 한국미생물·생명공학회지
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    • 제25권5호
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    • pp.468-476
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    • 1997
  • Soft-rot bacterial pathogen, Erwinia sp., was isolated from chinese cabbage tissue showing soft-rot symptom. This bacterial strain caused soft-rot to chinese cabbage and potato, and it was identified as Erwinia carotovora subsp. carotovora LY34 (E. c. subsp. carotovora LY34). Erwinia carotovora subsp. carotovora LY34 did not have hemicellulase but extracellular cellulase, pectinase, polygalacturonase, protease activity. The results of the microscopy showed that chinese cabbage tissue and potato tissue were macerated by infection of E. c. subsp. carotovora LY34. In analysis of the cellulases activity of the isolated cellulose-degradation enzymes from E. c. subsp. carotovora LY34 total protein, three cellulase activity bands were detected by non-denaturation gel electrophoresis method and five cellulase activity bands were detected by CMC-SDS-PAGE direct stain method.

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전종격동에 원발한 악성 흑색종 (Primary Malignant Melanoma Presenting as an Anterior Mediastinal Mass)

  • 김형렬;이정상;김영태;김주현;김지은
    • Journal of Chest Surgery
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    • 제35권9호
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    • pp.697-700
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    • 2002
  • 50세 여자가 전흉부의 통증을 주소로 내원하였다. 흉부단층촬영에서 우상엽을 침범하고 있는 전종격동의 종괴가 발견되었으며, 수술로서 완전절제하였다. 조직검사에서 악성 흑색종을 진단할 수 있었고, 원발병소의 가능성이 있는 피부, 구강, 안저, 비강, 직장, 항문, 질 등을 각각의 전문의에게 검사를 받았으나 이상소견이 없었다. 원발병소가 자연적으로 소멸된 경우가 아니라면 종격동에 원발한 악성 흑색종이라고 생각된다. 종격동에 발생하는 악성 흑색종은 아주 드문 질환으로 세계적으로 8례만이 보고되고 있고, 한국에서는 아직 예가 없기에 보고하는 바이다.

비중격에 발생한 신경초종 1례 (A Case of Neurilemmoma Arising from the Nasal Septum)

  • 조진희;장한성;김정묵;서병도
    • 대한기관식도과학회지
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    • 제5권1호
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    • pp.85-89
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    • 1999
  • Neurilemmoma is a benign, well-encapsulated neurogenic tumor that arises in cranial, peripheral, or autonomic nerves that have nerve sheaths containing schwann cells. From 16 to 45% of all neurilemmoma cases occur in the head and neck region and only about 4% of those cases are found in the nose and paranasal sinuses, but very rarely found in the nasal septum. Recently, we experienced a case of isolated neurilemmoma which arose from the nasal septum of a 30-year-old female patient who complained of nasal obstruction and postnasal drip. The mass was broadly based on the right anterior portion of the nasal septum and confined to the nasal cavity, displacing the lateral wall of the nasal cavity and middle turbinate, laterally and posteriorly, respectively. The tumor mass was completely excised through transnasal endoscopic surgery under general anesthesia. The final pathological diagnosis viewed under a microscope after H&E and anti S-100 protein stain was a benign neurilemmoma in which Antoni A type and B type both existed. This report presents a case of neurilemmoma which arose from the nasal septum with reviews of the clinicopathologic features.

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항결핵치료 종료후 발생한 농흉 (Empyema Occurred after Completion of Antituberculous Chemotherapy)

  • 윤기헌;유지홍;강홍모
    • Tuberculosis and Respiratory Diseases
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    • 제39권6호
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    • pp.554-558
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    • 1992
  • A 38 years old man had been treated as a pulmonary tuberculosis by the positive result of acid fast stain of bronchial washing from the focal infiltrative lesion at left lower lobe. On radiologic examination after one year treatment, there was an aggravation of lesion at left lower lobe with moderate amount of pleural effusion at the same side. After 11 weeks, follow up chest film disclosed bilateral pleural effusion. The pleural fluid of both side was pus in gross appearance with low pH, high LDH, low glucose and high protein. Pleurodectomy was performed to remove the loculated empyema with the thickened pleura of right thorax. This pleuro-pulmonary lesion can be easily misdiagnosed as a tuberculous lesion if it is not taken into consideration as a possible diagnosis.

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Bone Healing in Ovariectomized-rabbit Calvarial Defect with Tricalcium Phosphate Coated with Recombinant Human Bone Morphogenetic Protein-2 Genetically Engineered in Escherichia coli

  • Kim, Jung-Han;Kim, Chang-Joo;Shin, Sang-Hun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제36권2호
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    • pp.37-49
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    • 2014
  • Purpose: This study compares the bone formation ability of tricalcium phosphate (TCP) with and without recombinant human bone morphogenetic protein-2 (rhBMP-2) and assesses TCP as a carrier of rhBMP-2. Methods: Bilateral round defects (diameter: 8.0 mm) were formed in the cranium of eight New Zealand white rabbits. The defects were grafted with TCP only (control group) or with rhBMP-2-coated TCP (experimental group). The animals were sacrificed at 1st week, 2nd week, 4th week, and 8th week postoperatively; two rabbits sacrificed each time. The skulls were harvested and subjected to radiographic and histological examination. Results: Radiologic evaluation showed faster bone remodeling in the experimental group than in the control group. Histologic evaluation (H&E, Masson's trichrome stain) showed rapid bone formation, remodeling and calcification in the 1st and 2nd week in the experimental group. Immunohistochemical evaluation showed higher expression rate of osteoprotegerin, receptor activator of nuclear factor ${\kappa}B$ ligand, and receptor activator of nuclear factor ${\kappa}B$ in the experimental group at the 1st and 2nd week than in the control group. Conclusion: rhBMP-2 coated TCP resulted in rapid bone formation, remodeling, and calcification due to rhBMP-2's osteogenic effect. TCP performed properly as a carrier for rhBMP-2. Thus, the use of an rhBMP-2 coating on TCP had a synergic effect on bone healing and, especially, bone remodeling and maturation.

Anti-adipogenic effect of mulberry leaf ethanol extract in 3T3-L1 adipocytes

  • Yang, Soo Jin;Park, Na-Young;Lim, Yunsook
    • Nutrition Research and Practice
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    • 제8권6호
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    • pp.613-617
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    • 2014
  • BACKGROUND/OBJECTIVES: Adipogenesis is part of the cell differentiation process in which undifferentiated fibroblasts (pre-adipocytes) become mature adipocytes with the accumulation of lipid droplets and subsequent cell morphological changes. Several transcription factors and food components have been suggested to be involved in adipogenesis. The aim of this study was to determine whether mulberry leaf ethanol extract (MLEE) affects adipogenesis in 3T3-L1 adipocytes. MATERIALS/METHODS: The 3T3-L1 adipocytes were treated with different doses of MLEE for 8 days starting 2 days post-confluence. Cell viability, fat accumulation, and adipogenesis-related factors including CCAAT-enhancer-binding protein alpha ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), $PPAR{\gamma}$ coactivator 1 alpha (PGC-$1{\alpha}$), fatty acid synthase (FAS), and adiponectin were analyzed. RESULTS: Results showed that MLEE treatments at 10, 25, 50, and $100{\mu}g/ml$ had no effect on cell morphology and viability. Without evident toxicity, all MLEE treated cells had lower fat accumulation compared with control as shown by lower absorbances of Oil Red O stain. MLEE at 50 and $100{\mu}g/ml$ significantly reduced protein levels of $PPAR{\gamma}$, PGC-$1{\alpha}$, FAS, and adiponectin in differentiated adipocytes. Furthermore, protein level of $C/EBP{\alpha}$ was significantly decreased by the treatment of $100{\mu}g/ml$ MLEE. CONCLUSION: These results demonstrate that MLEE treatment has an anti-adipogenic effect in differentiated adipocytes without toxicity, suggesting its potential as an anti-obesity therapeutic.

무름병균 Erwinia chrysanthemi PY35의 CMCase isozymes 분리 (Isolation of CMCase Isozymes from Phytopathogenic Erwinia chrysanthemi PY35)

  • 박상렬;조수정;윤한대
    • Applied Biological Chemistry
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    • 제42권3호
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    • pp.199-204
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    • 1999
  • 배추로부터 연부균을 분리하여 조사한 결과 Erwinia chrysanthemi(Ech)로 동정되었으며 분리균 Ech PY35는 CMCase, pectinase, pectate lyase, protease 활성은 있었으나 hemicellulase 활성은 없었다. 배추와 감자의 조직에서 $24{\sim}48$시간 이내에 연부증상을 일으키며, 배추 조직보다 감자 조직에서 연부증상의 발병 진행속도가 느린 편이었다. 전자현미경관찰 결과 병원균의 식물 조직 내의 침투가 확인되었으며 이들이 분비한 세포벽분해효소에 의한 식물조직의 연부현상도 관찰되었다. 분리균의 체내 및 체외효소를 분리하여 CMC-SDS-PAGE에 의한 CMCase 직접활성 염색법을 이용하여 5종류의 CMCase isozyme으로 추정되는 활성밴드를 관찰할 수 있었다. 체내분비효소로 48 kDa, 34 kDa, 31 kDa의 3종류, 체외분비효소는 50 kDa, 29 kDa 2종류로 확인되었다.

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Antiobesity effects of the water-soluble fraction of the ethanol extract of Smilax china L. leaf in 3T3-L1 adipocytes

  • Kang, Yun Hwan;Kim, Kyoung Kon;Kim, Dae Jung;Choe, Myeon
    • Nutrition Research and Practice
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    • 제9권6호
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    • pp.606-612
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    • 2015
  • BACKGROUND/OBJECTIVES: Several medicinal properties of Smilax china L. have been studied including antioxidant, anti-inflammatory, and anti-cancer effects. However, the antiobesity activity and mechanism by which the water-soluble fraction of this plant mediates its effects are not clear. In the present study, we investigated the lipolytic actions of the water-soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) in 3T3-L1 adipocytes. MATERIALS/METHODS: The wsSCLE was identified by measuring the total polyphenol and flavonoid content. The wsSCLE was evaluated for its effects on cell viability, lipid accumulation, glycerol, and cyclic adenosine monophosphate (cAMP) contents. In addition, western blot analysis was used to evaluate the effects on protein kinase A (PKA), PKA substrates (PKAs), and hormone-sensitive lipase (HSL). For the lipid accumulation assay, 3T3-L1 adipocytes were treated with different doses of wsSCLE for 9 days starting 2 days post-confluence. In other cell experiments, mature 3T3-L1 adipocytes were treated for 24 h with wsSCLE. RESULTS: Results showed that treatment with wsSCLE at 0.05, 0.1, and 0.25 mg/mL had no effect on cell morphology and viability. Without evidence of toxicity, wsSCLE treatment decreased lipid accumulation compared with the untreated adipocyte controls as shown by the lower absorbance of Oil Red O stain. The wsSCLE significantly induced glycerol release and cAMP production in mature 3T3-L1 cells. Furthermore, protein levels of phosphorylated PKA, PKAs, and HSL significantly increased following wsSCLE treatment. CONCLUSION: These results demonstrate that the potential antiobesity activity of wsSCLE is at least in part due to the stimulation of cAMP-PKA-HSL signaling. In addition, the wsSCLE-stimulated lipolysis induced by the signaling is mediated via activation of the ${\beta}$-adrenergic receptor.

MSH에 의해 자극된 B16F10세포에서 사간(射干)의 멜라닌 합성 억제 효과 (Inhibitory Effect of Belamcandae Rhizoma on the Melanogenesis in MSH-stimulated B16F10 cells)

  • 김대성;성병곤;이장천;이부균;우원홍;임규상
    • 한방안이비인후피부과학회지
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    • 제24권1호
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    • pp.25-35
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    • 2011
  • Objective : The present study was designed to assess the potential inhibitory activity of an ethanol extract of Belamcandae Rhizoma (EBR) on the alpha-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis signal pathway in B16F10 melanoma cells. Methods : Several experiments were performed in B16F10 melanoma cells. We studied tyrosinase activity, melanin content, cell-free tyrosinase activity and DOPA stain, and performed Western blots and RT-PCR for proteins and mRNA involved in melanogenesis. Results : ${\alpha}$-MSH-induced tyrosinase activity and melanin content were inhibited significantly by EBR. EBR markedly suppressed the protein expression level of tyrosinase in B16F10 melanoma cells. On the other hand, the expression of tyrosinase-related protein-1 (TRP-1) and -2 (TRP-2; DCT) were not affected by EBR. To elucidate the mechanism of the depigmenting property of EBR, we examined the involvement EBR in cAMP response element binding (CREB) protein phosphorylation and microphthalmia-associated transcription factor (MITF) signalling induced by ${\alpha}$-MSH. EBR did not regulate CREB phosphorylation and MITF expression by ${\alpha}$-MSH. Nevertheless, the mRNA expression of tyrosinase was significantly attenuated by EBR treatment without changes in the expression of TRP-1 and -2 mRNA. Conclusion : Our study suggested that EBR inhibits ${\alpha}$-MSH-induced melanogenesis by suppressing tyrosinase mRNA.