• 제목/요약/키워드: Protein stain

검색결과 154건 처리시간 0.021초

Osteoblastogenesis and osteolysis in the Zucker Diabetic Sprague Dawley rat humerus head

  • Gcwalisile Frances Dlamini;Robert Ndou
    • Anatomy and Cell Biology
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    • 제56권4호
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    • pp.552-561
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    • 2023
  • The endocrinology of type 2 diabetes (T2D) and its predisposing factors have been studied extensively while its skeletal effects have received negligible research despite this being a global disease. The cellular and molecular association between proximal humeral fractures and T2D has not been fully elucidated. We aimed to study bone cell quantities and immunolabel osteogenic and antiosteogenic cytokines. The study used 12-week-old rats (23 males) consisting of 8 Sprague Dawley (SD) and 15 Zucker Diabetic Sprague Dawley (ZDSD). Weekly mass measurements were taken while fasting blood glucose levels were recorded every 2 weeks with oral glucose tolerance tests conducted once every 4 weeks. Upon termination at the age of 28 weeks, humeri were fixed in 10% buffered formalin, prior to decalcification in ethylenediaminetetraacetic acid. The bone samples were then processed in ascending grades of alcohol using an automatic processor before embedding in paraffin wax. Sections were cut at 5 ㎛ thickness in a series for Haematoxylin and Eosin stain, and immunohistochemistry was performed with the anti-tartrate-resistant acid phosphatase (TRAP), anti-alkaline phosphatase (ALP), anti-bone morphogenetic protein 3 (BMP3), anti-transforming growth factor beta 1 (TGFβ1), anti-aged glycation end product (AGE) antibodies in the sequence. ZDSD rats had more adipocytes, BMP3 and AGEs expression with higher numbers of TRAP positive osteocytes and fewer ALP cells although no differences were found in TGFβ1 immunopositivity. We also found that T2D increases the number of AGEs immuno-positive cells, as well as its extracellular expression, thus providing a conducive environment for the interaction of the osteogenic cytokine and its antagonist to suppress osteoblastogenesis. ZDSD groups had higher adipocyte numbers therefore increased marrow adiposity in T2D.

나물콩 재배시기가 종실 품질에 미치는 영향 (Effects of Sowing Date for Seed Quality of Sprout-soybean)

  • 김학신;김홍식;김경호
    • 한국작물학회지
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    • 제51권spc1호
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    • pp.152-159
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    • 2006
  • 본 연구는 나물용 콩의 파종기가 콩나물 원료콩의 품질에 미치는 영향을 구명함으로써 양질 콩나물 품종육성과 재배기술 확립을 위한 기초 자료를 얻고자 $2000{\sim}2001$년에 호남농업연구소 전작시험포장에서 파종시기 시험과 콩나물 간이검정실에서 콩나물생장 시험을 수행하였다. 시험 결과를 요약하면 다음과 같다. 1. 종실의 자반립, 열피립 및 미숙립 발생율은 5월 25일 파종이 6월 15일과 7월 5일 파종에 비하여 높았고, 풍산나물콩이 자반립, 열피립 및 미숙립 발생율이 가장 낮았으며 한남콩이 발생율이 가장 높았다. 2. 조단백질함량은 파종기가 늦어지면서 감소하였고, 5월 25일과 6월 15일 파종에서 다원콩이 가장 높았고 7월 5일 파종에서는 풍산나물콩이 가장 높았다. 조지방함량은 파종기에 따른 일정한 경향이 없었으며 소명콩이 가장 높았고 다원콩이 가장 낮았다. 3. Isoflavone의 aglycone 함량은 6월 15일 파종이 가장 많았고 다음으로 7월 5일파종이 많았으며, 5월 25일파종이 적었고, 품종간에는 소명콩이 가장 많았고 다원콩이 가장 낮았다. 4. 종실의 품질향상을 위한 나물콩의 파종기는 지나친 일찍 또는 늦게 심는 것 보다 품종 특성을 고려하여야 할 것으로 판단되었다.

Nitric Oxide-Induced Autophagy in MC3T3-E1 Cells is Associated with Cytoprotection via AMPK Activation

  • Yang, Jung Yoon;Park, Min Young;Park, Sam Young;Yoo, Hong Il;Kim, Min Seok;Kim, Jae Hyung;Kim, Won Jae;Jung, Ji Yeon
    • The Korean Journal of Physiology and Pharmacology
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    • 제19권6호
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    • pp.507-514
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    • 2015
  • Nitric oxide (NO) is important in the regulation of bone remodeling, whereas high concentration of NO promotes cell death of osteoblast. However, it is not clear yet whether NO-induced autophagy is implicated in cell death or survival of osteoblast. The present study is aimed to examine the role of NO-induced autophagy in the MC3T3-E1 cells and their underlying molecular mechanism. The effect of sodium nitroprusside (SNP), an NO donor, on the cytotoxicity of the MC3T3-E1 cells was determined by MTT assay and expression of apoptosis or autophagy associated molecules was evaluated by western blot analysis. The morphological observation of autophagy and apoptosis by acridine orange stain and TUNEL assay were performed, respectively. Treatment of SNP decreased the cell viability of the MC3T3-E1 cells in dose- and time-dependent manner. SNP increased expression levels of p62, ATG7, Beclin-1 and LC3-II, as typical autophagic markers and augmented acidic autophagolysosomal vacuoles, detected by acridine orange staining. However, pretreatment with 3-methyladenine (3MA), the specific inhibitor for autophagy, decreased cell viability, whereas increased the cleavage of PARP and caspase-3 in the SNP-treated MC3T3-E1 cells. AMP-activated protein kinase (AMPK), a major autophagy regulatory kinase, was activated in SNP-treated MC3T3-E1 cells. In addition, pretreatment with compound C, an inhibitor of AMPK, decreased cell viability, whereas increased the number of apoptotic cells, cleaved PARP and caspase-3 levels compared to those of SNP-treated MC3T3-E1 cells. Taken together, it is speculated that NO-induced autophagy functions as a survival mechanism via AMPK activation against apoptosis in the MC3T3-E1 cells.

의치 구내염 환자에서 분리한 Candida albicans의 아크릴 수지에 대한 부착성과 타액 단백질과의 상호 관계 (ADHESION OF CANDIDA ALBICANS ISOLATES TO ACRYLIC RESIN IN RELATION TO SALIVARY GLYCOPROTEINS IN DENTURE STOMATITIS PATIENTS)

  • 오정환;최부병;최대균;우이형;이성복;권긍록
    • 대한치과보철학회지
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    • 제37권5호
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    • pp.698-713
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    • 1999
  • Adherence of Candida albicans(C. albicans) to the surface of a denture is believed to be an initial and essential step in the formation of denture-induced stematitis. Previous studies have provided enormous infomation on the relationship between composition of palatine gland/parotid saliva and upper denture stomatitis. Relatively little information is available on the correlation between lower denture stomatitis and sublingual-submandibular ( SLSM ) saliva. The plaque samples were collected from the two sites($100mm^2$) on the inner surface of lower partial denture corresponding to the stematitis and healthy region of the lower partial dentures of 12 denture stomatitis patients and 6 nor-mal persons who wore lower partial dentures. The samples were plated to isolate C. albicans on a selective Saboraud's dextrose agar plate and the isolates were identified by germ tube test and gram staining. The subjects were divided into group I (stomatitis with C. albican), group II (lesion without C. albicans), group III (no lesion but C. albicans), and group IV (normal and healthy denture wearer). Individual SLSM saliva($20{\mu}g$ of protein) was analyzed by SDS-PAGE (SDS -poly-acrylamide gel electrophoresis) with Coomassie brilliant blue and PAS(Periodic Acid Schinff) stain-ing. The salivary proteins separated in the polyacryamide gels were subjected to immunoblot anaysis using anti-lactoferrin, anti-sIgA, and anti-secretory component of sIgA. In this study using custom made acrylic denture resin beads(5mm in diameter) coated with stimulated individual SLSM saliva, the binding ability of individual C. albicans strains to the beads was observed. Levels of C, albicans adhered to the acrylic resin beads were determined by measuring the optical density of the bound C. albicans to the beads at 580nm. The results showed that a higher number of C. albicans was observed in the lesion site than healthy site. The saliva of group I contained more high molecular weight glycoprotein(mucin, MGI) as compared to group II, III and IV. And lactoferrin and sIgA affected to the binding ability of C. albicans to acylic resin beads. Binding ability of individual C. albicans to the acrylic resin coated with respective individual saliva was found to be greater in group I than the other 3 groups. And when bound cells of C. albicans isolated from individual subject #2 to the saliva coated beads were used binding ability of subject #2 saliva coated beads was founed to be greater than the other sutjects. These results suggested that denture induced stomatitis is related to individual patient's salivary protein composition, especially MG-1. Future studies will be directed toward saliva exam-ination of patients who have general disease and analysis of pellicles formed on prosthesis with respect to oral disease.

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Lipo-PGE1의 투여경로가 흰쥐의 횡복직근피판의 생존률에 미치는 효과 (The Effect of Lipo-PGE1 According to the Routes of Administration on the Survival of Transverse Rectus Abdominis Musculocutaneous Flap in Rats)

  • 김성식;변준희;유결;한기택
    • Archives of Plastic Surgery
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    • 제32권1호
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    • pp.12-18
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    • 2005
  • The Transverse rectus abdominis musculocutaneous (TRAM) flap has been commonly used for autologous breast reconstruction. Despite these clinical usefulness, the TRAM flap is prone to partial flap or fat necrosis in especially pedicled flap. To improve flap survival, the surgical delay procedures and pharmacological treatments have been developed. In many studies for the pharmacological treatment, Lipo-$PGE_1$ has demonstrated a marked ability to improve flap survival and it's effect has been proved similar to surgical delay procedure. The purpose of this study is to determine the most effective route of Lipo-$PGE_1$ administration as a pharmacological treatment in TRAM flap of the rat. Fifty male Sprague-Dawley rats weighing 300-350 gm were divided into five groups, One week before flap elevation, Lipo-$PGE_1$($2{\mu}g/kg$) was injected three times in a week and than the left inferior epigastric vessel based TRAM flap ($5.0{\times}3.0cm$) elevated; group I: no procedure before flap elevation; group II: intraperitoneal injection; group III: intravenous injection; group IV: subcutaneous injection; group V: topical application. A flap was assessed at postoperative 7 days by comparison of flap survival rate, vessel counts(H-E stain), and vascular endothelial growth factor(VEGF) protein expressed by Western blot. The results demonstrated that the mean percentages of the flap survival area in group III were significantly higher than that of any other group(p<0.05). The vessel counts of all experimental groups were statistically higher than that of control group(p<0.05). Only in group III, the VEGF protein expression was increased significantly than control group and there are no difference in other experimental groups. In conclusion, the intravenous administration of the Lipo-$PGE_1$ is the most effective on flap survival, and the VEGF induced by Lipo-$PGE_1$ has some positive effects on new vessel formation and flap survival.

방사선 조사 후 생쥐(Mus musculus)조직의 형태와 젖산탈수소효소 동위효소의 재분포 (Redistribution of Lactate Dehydrogenase Isozymes and Morphology of Tissues in Mus musculus after Irradiation)

  • 박현도;염정주
    • 환경생물
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    • 제17권3호
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    • pp.263-270
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    • 1999
  • 생쥐에 1 Gray(Gy)와 3 Gy의 방사선을 조사한 후 시간에 따라 골격근, 심장, 신장, 간 및 정소조직의 형태적 변화를 확인하였다. H-B(hematoxylin-eosin)염색 결과 apoptotic body는 다른 조직에 비해 간조직에서 쉽게 관찰되었고, 조사량에 비례하였으며 대부분의 조직에서 조사 후 1일에, 정소조직에서는 7일에 많이 관찰되었다. TUNEL (terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling)염색에 의한 결과도 동일하게 나타났다. 방사선 조사 후 단백질량은 신장을 제외한 나머지 조직에서 감소되었으며, 3 Gy를 조사한 경우 1 Gy보다 모든 조직에서 초기 2시간까지 감소되었으나 7일에는 모두 증가되었다. 젖산탈수소효소(EC 1.1.1.27, Lactate dehydrogenase: LDH) 활성은 모든 조직에서 증가되었고 1 Gy를 조사한 경우 초기에 더욱 증가되었으며 최고 활성은 3 Gy보다 빠른 시기에 나타났다. 골격근, 심장, 신장 및 정소조직은 일반적으로 $A_4$동위효소 활성이 감소되고 골격근과 심장은 $B_4$$A_2$$B_2$동위효소 활성이 증가되었으며 신장은 이질사량체의 활성이 증가되었다. 간은 $A_4$ 동위효소 활성이 높았고 정소는 C동위효소의 활성이 높았다. 따라서 LDH는 방사선 조사시 간조직을 제외한 나머지 조직에서 lactate oxidase로서 작용하여 호기적 대사가 우세한 것으로 사료된다.

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다발성 폐종괴 및 기관지 점막으로 전이한 원발불명의 악성 흑색종 1예 (A Case of Unknown Primary Malignant Melanoma with Pulmonary and Endobronchial Metastasis)

  • 민영훈;김성욱;진희종;이태유;송헌호;이근석;이정애;박영이;현인규
    • Tuberculosis and Respiratory Diseases
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    • 제53권2호
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    • pp.196-201
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    • 2002
  • 악성 흑색종은 멜라닌세포에서 발생하는 종양으로 전체 악성 종양의 3%를 차지하고, 피부, 안구, 피부주위점막에서 호발한다. 이들은 주로 피부, 림프절, 폐, 간, 골 등으로 전이되고 폐의 전이는 약 5~15%에서 일어난다. 그러나 기관지 점막으로의 전이는 흔하지 않다. 특히 기관지 및 폐장으로 전이된 원발불명의 악성 흑색종은 아직 보고된 예가 없다. 이에 저자들은 기침과 검은색 객담을 주소로 내원하였던 34세 남자환자에서 굴곡성 기관지 내시경 및 조직검사를 시행하여 vimentin, S-100 protein, HMB-45 염색에 양성을 보이는 양측 폐장 및 기관지점막으로 전이된 악성 흑색종을 경험하였기에 문헌고찰과 함께 보고하는 바이다.

편평세포폐암에서 Argyrophilic Nucleolar Organizer Regions(AgNORs)의 임상적의의 (Clinical Significance of Argyrophilic Nucleolar Organizer Regions(AgNORs) In Squamous Cell Carcinoma of the Lung)

  • 한승범;전영준;이상숙
    • Tuberculosis and Respiratory Diseases
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    • 제42권4호
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    • pp.513-521
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    • 1995
  • 연구배경: 세포의 증식능력을 반영한다고 알려진 NORs를 간편한 은염색으로 발현시켜서 AgNORs 수가 정상조직과 종양조직의 비교감별과 종양의 증식능력판단에 유용한지를 알기위하여 파라핀 포매된 편평세포폐암 조직을 이용하여 본 연구를 시행하였다. 방법: 수술로써 절제한 파라핀 포매된 편평세포폐암 조직 36예를 Mourad 등의 방법으로 은염색하였다. 1,000 배 배율하에서 종양조직과 정상조직에서 임의로 100 개씩 세포를 선정해서 핵당 평균 AgNORs수를 구하였다. 결과: 종양조직에서 TNM병기에 따른 핵당 평균 AgNORs수는 유의한 차이가 없었으나 정상조직과 종양조직을 비교허여 정상조직에서는 $1.74{\pm}0.25$, 종양 조직에서는 $4.05{\pm}0.80$으로 종양조직의 핵당 평균 AgNORs수가 정상조직에 비해서 유의하게 높았다(p<0.001). 결론: TNM분류에 따른 각 병기별 종양조직과 인접 정상조직의 핵당 평균 AgNORs수를 비교하여도 각 병기에서 종양조직의 평균 AgNORs수가 정상조직에 비하여 유의하게 높았다(p<0.005).

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상아모세포의 조건배지를 이용한 백악모세포의 분화와 석회화 조절 (Regulation of cementoblast differentiation and mineralization using conditioned media of odontoblast)

  • 문상원;김혜선;송혜정;최홍규;박종태;김흥중;장현선;박주철
    • Journal of Periodontal and Implant Science
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    • 제36권2호
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    • pp.385-396
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    • 2006
  • For the regeneration of periodontal tissues, the microenvironment for new attachment of connective tissue fibers should be provided, At this point of view, cementum formation in root surface plays a key role for this new attachment. This study was performed to figure out which factor promotes differentiation of cementoblast Considering anatomical structure of tooth, we selected the cells which may affect the differentiation of cementoblast - Ameloblast, OD11&MDPC23 for odontoblasts, NIH3T3 for fibroblsts and MG63 for osteoblasts. And OCCM30 was selected for cementoblast cell line. Then, the cell lines were cultured respectively and transferred the conditioned media to OCCM30. To evaluate the result, Alizarin red S stain was proceeded for evaluation of mineralization. The subjected mRNA genes are bone sialoprotein(BSP), alkaline phosphate(ALP) , osteocalcin(OC), type I collagen(Col I), osteonectin(SPARC ; secreted protein acidic and rich in cysteine). Expression of the gene were analysed by RT-PCR, The results were as follows: 1. For alizarin red S staining, control OCCM30 didn't show any mineralized red nodules until 14 days. But red nodules started to appear from about 4 days in MDPC-OCCM30 & OD11-OCCM30. 2. For results of RT-PCR, ESP mRNAs of control-OCCM30 and others were expressed from 14 days, but in MDPC23-OCCM30 & OD11-OCCM30 from 4 days. Like this, the gene expression of MDPC23-OCCM30 & OD11-OCCM30 were detected much earlier than others. 3. For confirmation of odontoblast effect on cementoblast, conditioned media of osteoblasts(MG63) which is mineralized by producing matrix vesicles didn't affect on the mineralized nodule formation of cementoblasts(OCCM30). This suggest the possibility that cementoblast mineralization is regulated by specific factor in dentin matrix protein rather than matrix vesicles. Therefore, we proved that the dentin/odontoblast promotes differentiation/mineralization of cementoblasts. This new approach might hole promise as diverse possibilities for the regeneration of tissues after periodontal disease.

방사선에 의한 폐 섬유화증에서 c-Jun N-terminal Kinase(JNK)의 역할 (The Role of c-Jun N-terminal Kinase in the Radiation-Induced Lung Fibrosis)

  • 어수택;홍기영;이영목;김기업;김도진;;김용훈;박춘식;염욱;김은석;최두호
    • Tuberculosis and Respiratory Diseases
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    • 제50권4호
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    • pp.450-461
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    • 2001
  • 서 론 : 폐암의 치료에 사용되는 방사선 조사는 흉곽 및 여러장기에 다양한 합병증을 발생시키며, 특히 방사선 섬유화증과 방사선 폐렴을 일으킨다. 방사선 조사의 초기 효과는 보통 염증 세포들의 침윤, 간질 및 폐포 부종, 상피세포의 탈락에 의한 방사선 폐렴이며 후기 효과는 방사선 섬유화증을 초래한다. 방사선 조사는 폐장의 염증 세포에서 TGF-beta의 단백 합성 및 활성도를 증가시키고, 생체외 실험에서 TGF-beta는 mitogen activated protein kinases(MAPKs)를 활성화시킨다는 것이 알려져 있다. c-Jun N-terminal kinase(JNK)는 MAPKs중의 하나로 핵단백질인 c-Jun을 인산화(phosphorylation) 시켜 전사(transcription)를 증가시키는데 생체외 실험에서 자외선 조사후 대식 세포에서 JNK의 활성이 증가되는 것으로 알려져 있다. 하지만 현재 까지 생체내에서 JNK가 방사선 조사에 의해 활성화되는지 그리고 이들 활성화가 방사선 섬유화증의 병인에 관계하는 지는 알려진 바 없다. 본 연구는 흉부에 방사선을 조사한 백서를 이용하여 방사선 섬유화증의 병인에 JNK가 신호 전달 체계에서 주요한 역할을 담당하는 지를 알아보고자 시행하였다. 대상 및 방법 : C57BL/6 백서의 전 흉부에 14 Gy의 $^{60}CO{\gamma}$-ray를 조사한 후 일정한 간격(1주, 4주, 8주)으로 폐장의 세포 분석을 위한 기관지 폐포 세척술, elastin의 합성 정도를 측정하기 위한 Verhoeff stain, collagen 합성 양을 알기 위한 hydroxyproline의 측정, JNK 활성도를 알기 위한 in vitro JNK assay를 시행하였다. 결 과 : 폐장의 기관지 폐포 세척 소견상 총세포수는 방사선 조사 4주, 8주후에 증가하는 소견을 보였다. 세포의 감별 분석상 림프구는 4주후 증가되는 경향을 보였다. Verhoeff 염색상 폐포 조직의 특이 변화 소견은 없었으며 hydroxyproline의 양은 방사선 조사전에 비해 4주, 8주후에 증가하는 소견을 보였다. 방사선 조사 후 시간이 경과함에 따라 4주 후에 c-Jun N-terminal kinase(JNK)의 활성도가 가장 높았으며 8주 후에는 4주 후와 비슷한 활성도를 보였다. 결 론 : 흉곽에 방사선 조사 후 hydroxyproline양의 증가와 함께 JNK 활성도의 증가 소견을 보여 방사선 폐섬유화증의 병인에 JNK가 관여될 것으로 사료된다.

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