• Title/Summary/Keyword: Protein stain

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THE EFFECT OF HYALURONIC ACID ON MOUSE CALVARIA PRE-OSTEOBLASTS OSTEOGENESIS IN VITRO (히알루론산이 골 형성에 미치는 영향에 관한 실험적 연구)

  • Cho, Yong-Min;Min, Seung-Ki;Kim, Soo-Nam;You, Yong-Ouk
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.3
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    • pp.216-225
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    • 2002
  • Hyaluronic acid (HA) is an almost essential component of extracellular matrices. Early in embryogenesis mesenchymal cells migrate, proliferate and differentiate, in part, because of the influence of HA. Since the features of embryogenesis are revisited during wound repair, including bone fracture repair, this study was initiated to evaluate whether HA has an effect on calcification and bone formation in an in vitro system of osteogenesis. Mouse calvaria Pre-osteoblast (MC3T3-E1) cells were cultured in ${\alpha}-MEM$ medium with microorganism-derivative hyaluronic acid that was produced by Strep. zooepidemicus which of molecular weight was 3 million units. The dosages were categorized in each 0.5, 1.0 and 2.0 mg/ml concentration experimental groups. After 2 and 4 days cultures in expeirmental and control groups, the tendency of cell proliferation, MTT assay, protein synthesis ability, collagen synthesis and alkaline phosphatase activity were analysed and bone nodule formation capacity were measured with Alizarin Red S stain after 29 days cultures. The cell proliferation was increased in time, especially the group of 0.5 and 1.0 mg/ml concentration of HA were showed prominent cell proliferation. After 2 and 4 days culture, experimental groups in general were greater cell activity in MTT assay. The protein synthesis was increased in all experimental groups compared to control group, especially most prominent in 1.0 mg/ml concentration group. The collagen synthesis capacity were increased in HA experimental groups, especially prominent in 1.0 mg/ml group and the activity of alkaline phosphatase were increased, especially also prominent in 1.0 mg/ml group, compared to control group. Above these, the activity of mouse carvarial pre-osteoblast cells was showed greater bone osteogenesis activity in all applied HA experimental group, especially group of 1.0 mg/ml concentration of HA.

Electrophoretic analysis of the major proteins of race horse erythrocyte membrane: Their relation to fast erythrocyte sedimentation rate (경주 적혈구막 단백의 전기영동법에 의한 분석 -빠른 적혈구 침강 속도와의 관계-)

  • Bahk, Yeong-woo;Lee, Si-yeong
    • Korean Journal of Veterinary Research
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    • v.31 no.3
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    • pp.259-264
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    • 1991
  • The proteins of the race horse erythrocyte membrane were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate(SDS-PAGE), and their relations to the fast erythrocyte sedimentation rate(ESR) of the race horse were investigated. The erythrocyte sedimentation rate of race horse were very fast compared with the human one(33 times <$90^{\circ}-plastic-ESR/30m$> and 25 times <$90^{\circ}-micro-ESR/30m$> as fast as the human one) are reported previously. Although the general protein profiles of the race horse erythrocyte membranes were almost similar to that of human, band 3 content was showing higher in race horse (34.7%) than in human (25.3%). The glycoprotein profiles of the race horse erythrocyte membranes revealed by periodic acid Schiff's(PAS) stain showed a marked difference from that of human. The PAS-1(glycophorin) and PAS-2(sialoglycoprotein) present in human erythrocyte memo brane were almost absent from the Holstein and race horse erythrocyte membranes, but PAS-2 was more in only race horse from that of human. Instead, the bovine erythrocyte membranes showed a strong PAS-B near the origin of the electrophorograms and the race horse erythrocyte membranes showed a strong PAS-negative band near the end of the electrophorograms, which is named as PAS-E in this study. These results suggest that the fast sedimentation rate of race horse erythrocyte is due in part to the presence of more band 3 protein fraction and PAS-E glycoproteins in the race horse erythrocytes.

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Inhibitory Effect of Ethyl Acetate Extract of White Peach Pericarp on Adipogenesis of 3T3-L1 Preadipocyte Cells

  • Park, Hong-Gyu;Kim, Jin-Moon;Kim, Jung-Mogg;Chung, Won-Yoon;Yoo, Yun-Jung;Cha, Jeong-Heon
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1327-1331
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    • 2008
  • In order to determine whether peach contains compounds to regulate adipocyte differentiation, extracts of flesh/pericarp of yellow/white peach were prepared in water, ethyl acetate (EtOAc), or n-butanol solvent and determined for effects on adipocyte differentiation in C3H10T1/2 or 3T3-L1 cells. Interestingly, none of peach extracts has statistically significant stimulatory effect on the adipocyte differentiation in C3H10T1/2. Furthermore, the presence of EtOAc extract of white peach pericarp (WPP) was found to inhibit lipid accumulation in 3T3-L1 cells both by microscopic examination of Oil Red O-stained lipid droplets and by spectrophotometric quantification of extracted stain, indicating a significant inhibitory effect on adipocyte differentiation. The inhibition of lipid accumulation was accompanied by a significant decrease in the expression levels of adipocyte molecular markers-peroxisome proliferator-activated receptor $\gamma$, CAAT enhancer binding protein $\alpha$, and fatty acid-binding protein. Thus, this study determined that WPP EtOAc extract contains the inhibitory compound(s) on adipogenesis.

Fine Needle Aspiration Cytology of Cervical Chordoma - A Case Report - (경추에 발생한 척삭종의 세침흡인 세포학적 소견 - 1예 보고 -)

  • Yang, Kyung-Moo;Cho, Mee-Yon;Jung, Soon-Hee;Bong, Jeong-Pyo
    • The Korean Journal of Cytopathology
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    • v.8 no.1
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    • pp.93-97
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    • 1997
  • A case of cervical chordoma diagnosed by fine needle aspiration is discussed. A 41year-old male was admitted due to dyspnea on neck flexion. Radiologic image revealed a retrotracheal superior mediastinal solid mass. Aspiration cytology showed many clusters of oval or large polygonal cells having abundant eosinophilic or bubbly cytoplasm in an amorphous blue-gray mucoid background. The nuclei were round and showed size variation, coarse granular chromatin, and indistinct nucleoli. Some cells contained brown granular pigments in the cytoplasm. Mitoses were rarely found. The cytoplasm was strongly positive for PAS stain. Immunohistochemical stains using cell block revealed positive reaction for cytokerain, EMA, vimentin, and S-100 protein. The confirmative diagnosis was made by following excisional biopsy. Electron microscopic study revealed large pools of intracytoplasmic glycogen and microfilaments. This is the first case of cervical chordoma diagnosed by aspiration cytology to our knowledge in Korean literature.

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RASA1-Related Parkes Weber Syndrome in a Neonate

  • Koh, Hong Ryul;Lee, Yeon Kyung;Ko, Sun Young;Shin, Son Moon;Han, Byoung-Hee
    • Neonatal Medicine
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    • v.25 no.3
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    • pp.126-130
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    • 2018
  • Parkes Weber syndrome is a rare congenital vascular anomaly, related to the RAS p21 protein activator 1 (RASA1) gene. It is characterized by capillary cutaneous malformations, bony and soft tissue hyperplasia, and multiple arteriovenous fistulas throughout the affected upper or lower extremity. These arteriovenous fistulas can be associated with life-threatening complications such as bleeding, thrombosis, and high output heart failure. In this report, we present a neonate who had a disproportionately hypertrophied left upper limb with port-wine stain, dystrophy of the left humerus, and hypertrophy of the left clavicle on X-ray, and arteriovenous malformation and massive dilatation of the left subclavian artery on magnetic resonance angiography. Exome sequencing analysis revealed a novel heterozygous splicing mutation (c.1776+2T>A) in the RASA1 gene. To the best of our knowledge, this report is the first case of RASA1-related Parkes Weber syndrome in Korea.

Cytologic Findings of Epithelial - Myoepithelial Carcinoma of the Salivary Gland - A Cese Report - (타액선 상피-근상피세포암종의 세포학적 소견 - 1예 보고 -)

  • Nam, Eun-Sook;Kang, Gu;Shin, Hyung-Sik
    • The Korean Journal of Cytopathology
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    • v.7 no.1
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    • pp.64-68
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    • 1996
  • The report of aspiration cytologic findings of epithelial-myoepithelial carcinoma(EMC) in the salivary gland is extremely rare. We present a case of fine needle aspiration cytology(FNAC) from EMC in the right submandibular gland of a 46 years old male patient. Neck CT scan revealed a confined lesion in the submandibular gland without enlargement of the regional lymph node. FNAC from the tumor showed several three-dimensional cellular clusters with admixed normal acinar cells. They frequently formed blanching tubular structures composed of two type of cells; darker cells haying eosinophilic scanty cytoplasm with round dense nuclei and clear cells having abudant pale cytoplasm with vesicular nuclei at the periphery of clusters. The tumor cells of both types did not show pleomorphism or mitoses. The resected submandibular gland showed an ill-defined whitish firm tumor, measuring $2{\times}1.5{\times}2cm$. The histology revealed an infiltrative tumor showing characteristic two cell types in a duct-like arrangement surrounded by thin basement menbrane. An inner layer of darker cells and outer layer of clear cells were postive for cytokeratin in the former and S-100 protein in the taller on the immunohistochemical stain.

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[ $PLC-{\gamma}1$ ] for Differentiating Adenocarcinoma from Reactive Mesothelial Cells in Effusions (체강 삼출액의 진단에 있어서 $PLC-{\gamma}1$ 면역 염색의 유용성)

  • Woo, Yeong-Ju;Kim, Sung-Sook
    • The Korean Journal of Cytopathology
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    • v.8 no.2
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    • pp.115-119
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    • 1997
  • Cytologic diagnosis of reactive or malignant effusion is sometimes difficult. Especially, differentiation of benign reactive mesothelial cells from malignant cells in body effusion is more difficult. Recently, immunohistochemistry has been used to diagnose difficult cases. Phospholipase $C(PLC)-{\gamma}1$ is one of the isoenzyme of the PLC which plays central role in signal transduction involving cellular growth, differentiation and transformation by phosphorylating many protein component. Increased expression of $PLC-{\gamma}1$ in human breast carcinoma, colorectal carcinoma and stomach cancers are reported. To evaluate the efficacy of positive $PLC-{\gamma}1$ immunostaining in the diagnosis of malignancy in effusions, paraffin-embedded cell blocks of pleural fluid and ascites from 10 patients(5 metastatic adenocarcinomas, and 5 reactive mesothelial cells) were immunostained with a monoclonal antibody to $PLC-{\gamma}1$. $PLC-{\gamma}1$ immuostained all the adenocarcinomas in cell block(5/5) with intense membrane pattern, however, none of the reactive mesothelial proliferations stained with the diagnostic membrane pattern. Thus, our study strongly supports the conclusion that $PLC-{\gamma}1$ immunopositivity is likely to become a useful adjunct for the diagnosis of malignancy in effusions.

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Effect of Ethanol on Mouse Brain Cell

  • Jang, Hyung Seok
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.1
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    • pp.51-58
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    • 2015
  • Ethanol has long been implicated in triggering apoptotic neurodegeneration. Alcohol also may indirectly harm the fetus by imparing the mother's physiology. We examined the effects of ethanol on immature brain of mice. Three-weeks-old female ICR strain mice daily intraperitoneally injected with ethanol at the concentration of 4 and 20% in saline for 0, 6, and 24 hours and 1 and 4 weeks. The mice were weighted and sacrificed, and the brains were ectomized for the present histological, immunohistochemical and TUNEL assays. Based on the histologic hematoxylin and eosin stain, immunohistochemical expression of glutamate receptor protein and neuronal cell adhesion molecule (NCAM) were evaluated. The cerebral cortex of the ethanol-treated group showed few typical symptoms of apoptosis such as chromosome condensation and disintegration of the cell bodies. TUNEL staining revealed DNA fragmentation in the 6 and 24 hours. This results demonstrated that acute ethanol administration causes neuronal cell death. I found that either glutamate receptor inhibition or activation could induce cerebellar degeneration as ethanol effect. Neuronal death also can be induced by excess activity of certain neurotransmitter, including glutamate. Neurons must establish cell-to-cell contact during growth and development in order to survive, migrate to their final destination, and develop appropriate connections with neighboring cell. Purkinje cell in cerebellar are especially vulnerable to the cell death and degeneration. After ethanol treatment in cerebellar, NCAM had decreased by 4 weeks. This result suggest that apoptosis seems to be involved in the slow elimination of neuron and cerebellar degeneration.

Effects of Hwangryunhaedoktang on DSS-induced Colitis (황련해독탕이 DSS로 유발된 흰쥐의 궤양성 대장염에 미치는 영향)

  • An Joong-Hwan;Choi En-Young;Lee Seong-Hwan;Lee Seong-Hwan;Lim Seong-Woo
    • The Journal of Korean Medicine
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    • v.27 no.2 s.66
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    • pp.182-195
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    • 2006
  • Objectives : We examined the effect of Hwangryunhaedoktang(HH) on the experimental ulcerative colitis induced by dextran sulfate sodium (DSS). Methods : Experimental colitis was induced in rats by daily treatment with 5% DSS in the drinking water for 5 days. Afterward, the mts were divided into two groups: the control group was administered water and the sample group was administered HH for 7 days. Results : The sample group provided HH for 7 days demonstrated fast recovery of body weight compared with the control group. Histologic change showed fast regeneration of crypt and surface epithelial cells and decreased edema of the submucosa and decreased lymphatic follicle of mucosa compared with the control group. Immunohistochemical stain usingCOX-2 gene was decreased and there was localized Ki-67 positive reaction. Regeneration of surface epithelial cell and goblet cell in mucosa was observed by transmission electron microscope. These results indicate therapeutic effect of HH on DSS-induced colitis in rats.

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Molecular Association of Glucose Transporter in the Plasma Membrane of Rat Adipocyte

  • Hah, Jong-Sik
    • The Korean Journal of Physiology
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    • v.25 no.2
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    • pp.115-123
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    • 1991
  • Molecular association of glucose transporters with the other proteins in the plasma membrane was assessed by gel electrophoresis and immunoblot techniques. Approximately $31.5{\pm}5.1%$ of GLUT-4, $64.8{\pm}2.7%$ of clathrin, 48.7% of total protein in the plasma membrane (PM) were found insoluble upon extraction with 1% Tx-100. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the Tx-100 insoluble PM fraction contained about 4 major polypeptides with apparent molecular weight of above 200, 100-120, 80 and 30-35 KDa that were readily removed upon wash with a high pH buffer which is known to remove clathrin and 0.5 M Tris-buffer which is known to remove assembly proteins (AP). Immunoblotting of GLUT4 and clathrin against specific antibodies showed that GLUT-4 and clathrin were co-solubilized up to 84.6% and 82.7% respectively by wash with a high pH buffer and 1% Tx-100. When the membrane was pre-washed with a high pH buffer and 0.5 M Tris solution, GLUT4 and clathrin were not solubilized further suggesting that GLUT4 molecules are in molecular association with clathrin, AP and/or other extrinsic membrane proteins in plasma membrane and the formation of clathrin-coated structures might be involved in insulin stimulated glucose transporter translocation mechanism.

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