• 제목/요약/키워드: Cell labeling

검색결과 315건 처리시간 0.023초

Progression of Apoptotic Cells by Pretreatment of Proteinase K

  • Joo, Kyeng-Woong
    • 대한의생명과학회지
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    • 제8권3호
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    • pp.161-165
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    • 2002
  • Apoptosis can be difficult to detect in routine histological sections. Since extensive DNA fragmentation is an important characteristic of this process, visualization of DNA breaks could greatly facilitate the identification of apoptotic cells. Several techniques for the qualitative and quantitative detection of this process have been established; recently, an in situ nick end-labelling technique based on the detection of DNA fragmentation, which is a molecular characteristic of apoptotic cell death, was described. Applying this method to paraffin sections of rat tissues, sensitivity was observed to be inconsistently low with regard to the expected number of apoptotic cells. I describe a new modified method for formalin-fixed, paraffin-embedded tissue sections, pretense pretreatment to permeate the tissue sections that involves an TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling) is acknowledged as a method of choice in the rapid identification and quantification of the apoptotic cell fraction in paraffin tissue preparations. TUNEL was performed without apoptosis and with apopotosis samples to each of the three concentrations of proteinase K (10, 25, 40 mg/ml) pretreatments. In this study, I show that chemical pretreatments of the tissue sections in proteinase K (25 mg/ml for 15 min at room temperature) considerably enhances the sensitivity of this nick end labelling technique.

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Protective Effects of Singihwan (腎氣丸) on Traumatic Brain Injury-induced Apoptosis in Rat Hippocampal Dentate Gyrus

  • Kwon, Oh-Bong;Song, Yun-kyung;Lim, Hyung-Ho
    • 대한한의학회지
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    • 제29권2호
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    • pp.21-31
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    • 2008
  • Backgrounds: Singihwan is used "to strengthen inborn energy" and we suspected a protective effect on brain neuron cells. Objectives: The aim of this study was to evaluate the effects of Singihwan (SGH) on traumatic brain injury-induced delayed apoptosis in rat hippocampal dentate gyrus. Methods: For a surgical induction of traumatic brain injury (TBI), a 5 mm diameter stainless rod was used to make traumatic attack from the surface of the brain used by an impactor. The protective effect of the aqueous extract of SGH against TBI in the rat hippocampal dentate gyrus was investigated by using step-down avoidance task, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, Bax immunohistochemistry, and 5-bromo-2'-deoxyuridine (BrdU) immunohistochemistry. Results: The aqueous extract of SGH suppressed the TBI-induced increase in apoptosis and cell proliferation in the hippocampal dentate gyrus. Conclusions: It is possible that the aqueous extract of SGH has a neuroprotective effect on TBI-induced neuronal cell death.

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Morphological Feature Extraction of Microorganisms Using Image Processing

  • Kim Hak-Kyeong;Jeong Nam-Su;Kim Sang-Bong;Lee Myung-Suk
    • Fisheries and Aquatic Sciences
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    • 제4권1호
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    • pp.1-9
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    • 2001
  • This paper describes a procedure extracting feature vector of a target cell more precisely in the case of identifying specified cell. The classification of object type is based on feature vector such as area, complexity, centroid, rotation angle, effective diameter, perimeter, width and height of the object So, the feature vector plays very important role in classifying objects. Because the feature vectors is affected by noises and holes, it is necessary to remove noises contaminated in original image to get feature vector extraction exactly. In this paper, we propose the following method to do to get feature vector extraction exactly. First, by Otsu's optimal threshold selection method and morphological filters such as cleaning, filling and opening filters, we separate objects from background an get rid of isolated particles. After the labeling step by 4-adjacent neighborhood, the labeled image is filtered by the area filter. From this area-filtered image, feature vector such as area, complexity, centroid, rotation angle, effective diameter, the perimeter based on chain code and the width and height based on rotation matrix are extracted. To prove the effectiveness, the proposed method is applied for yeast Zygosaccharomyces rouxn. It is also shown that the experimental results from the proposed method is more efficient in measuring feature vectors than from only Otsu's optimal threshold detection method.

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적포도의 주 항산화물질, 레스베라트롤의 항암작용: 아폽토시스 의한 인체 암세포 사멸 유도 (Anticarcinogenic Activity of Resveratrol, a Major Antioxidant Presnet in Red Wine : Induction of Apoptosis in Human Cancer Cells)

  • 허연진;김정환;서효정;공구;서영준
    • 한국환경성돌연변이발암원학회지
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    • 제19권1호
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    • pp.56-62
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    • 1999
  • Resveratrol (3,5,4'-trihydroxy-trans-stilbene) has been considered to be as one of major antioxidants present in grapes responsible for beneficial effects of red wine consumption on coronary heart disease. This triphenolic stilbene has been suggested as a potential cancer chemopreventive agent based on its striking inhiitory effects on diverse cellular events associated with tumor initiation, promotion, and progression. The compound has strong antioxidative and anti-inflammatory activities which amy contribute to its chemopreventive/chemoprotective properties. In the present work, we have found that resveratrol reduces viability and DNA synthesis capability of cultured human promyelocytic leukemia (HL-60) cells. Likewise, the viability of human breast cancer cell line, MCF-7 was reduced by resveratrol treatment. The growth inhibitory and antiproliferative properties of resveratrol appear to be associated with its induction of apoptotic cell death as determined by morphological and ultrastructural changes, agarose gel electrphoretic analysis of internucleosomal DNA fragmentation, and in situ terminal end-labeling of fragmented DNA (TUNEL). This compound also inhibited the phorbol ester-induced expression of cyclooxygenase-2 (COX-2) protein in immortalized human mammary epithelial MCF-10A cells. These results suggest that resveratrol has the promising cancer therapeutic/chemopreventive potential.

가미팔미환(加味八味丸)의 elastase 유도성 A549 세포사멸에 대한 보호효과 (Protective Effects of Gamipalmi-hwan on Elastase-induced Apoptosis of A549 Cells)

  • 오지석;박양춘
    • 대한한의학회지
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    • 제31권2호
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    • pp.137-148
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    • 2010
  • Objective: This study aimed to evaluate the protective effects of Gamipalmi-hwan (GPH) on elastase-induced lung cell injury. Materials and Methods: As an in vitro model of emphysema, the current study was performed to investigate potential activity of GPH in regulating injury responses of A549 human type II cell line mediated by elastase treatment. Results: GPH treatment increased the number of A549 cells which was reduced by elastase digestion. Elastin protein level, which was reduced by elastase treatment, was increased by GPH treatment. Labeling intensity with caspase 3 protein in elastase-treated cells was reduced by GPH treatment. Both Erk1/2 and Cdc2 protein levels, which were decreased by elastase treatment, were increased to a level similar to that of the normal cells. mRNA levels encoding IL-$1{\beta}$ and TNF-$\alpha$ were increased by elastase and then down-regulated by GPH. Conclusion: The present data suggest that A549 cells are subjected to inflammatory damage by elastase and can be recovered by GPH treatment. Further studies examining the protective activity of GPH in elastase-treated lung tissue would be useful for therapeutic strategies of emphysema treatment.

Spindle cell myoepithelioma of the parotid gland

  • Oh, Suk Joon;Moon, Dukju
    • 대한두개안면성형외과학회지
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    • 제20권5호
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    • pp.336-340
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    • 2019
  • Myoepithelioma was recognized as a histological distinct entity by the World Health Organization (WHO) in 1991. Myoepithelial cells are believed to be of ectodermal origin. In salivary glands, the myoepithelial cells that surround the intercalated ducts are spindled, which is in contrast to the large stellate ones that envelop the acini. Myoepithelioma is a benign salivary gland tumor that consists entirely of myoepithelial cells. A 53-year-old man presented with a 1-year history of a painless mass originating from the right parotid gland. The mass grew rapidly reaching a size of approximately 6 cm. The patient had no facial paralysis. The authors performed right parotidectomy. Immunohistochemistry study of this tumor showed that it was positive for vimentin, positive for S-100, focally positive for pancytokeratin, and focally positive for p63 and that it had a Ki-67 labeling index (below 10%). Additionally, the tumor was negative for epithelial membrane antigen, negative for actin, negative for desmin, negative for CD34 and negative for anaplastic lymphoma kinase. The authors present a case of benign spindle cell myoepithelioma of the parotid gland in a 53-year-old man diagnosed after immunohistochemistry study, describing its importance, along with a brief review of the literature.

Calnexin as a dual-role biomarker: antibody-based diagnosis and therapeutic targeting in lung cancer

  • Soyeon Lim;Youngeun Ha;Boram Lee;Junho Shin;Taiyoun Rhim
    • BMB Reports
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    • 제57권3호
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    • pp.155-160
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    • 2024
  • Lung cancer carries one of the highest mortality rates among all cancers. It is often diagnosed at more advanced stages with limited treatment options compared to other malignancies. This study focuses on calnexin as a potential biomarker for diagnosis and treatment of lung cancer. Calnexin, a molecular chaperone integral to N-linked glycoprotein synthesis, has shown some associations with cancer. However, targeted therapeutic or diagnostic methods using calnexin have been proposed. Through 1D-LCMSMS, we identified calnexin as a biomarker for lung cancer and substantiated its expression in human lung cancer cell membranes using Western blotting, flow cytometry, and immunocytochemistry. Anti-calnexin antibodies exhibited complement-dependent cytotoxicity to lung cancer cell lines, resulting in a notable reduction in tumor growth in a subcutaneous xenograft model. Additionally, we verified the feasibility of labeling tumors through in vivo imaging using antibodies against calnexin. Furthermore, exosomal detection of calnexin suggested the potential utility of liquid biopsy for diagnostic purposes. In conclusion, this study establishes calnexin as a promising target for antibody-based lung cancer diagnosis and therapy, unlocking novel avenues for early detection and treatment.

Cyclohexanone의 in vivo 소핵시험을 통한 GHS 변이원성 구분 (In vivo Micronucleus Test of Cyclohexanone and Mutagenicity Classification According to a Globally Harmonized System)

  • 김수진;임경택;임철홍
    • 생명과학회지
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    • 제24권7호
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    • pp.804-811
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    • 2014
  • Cyclohexanone의 GHS 분류 기준에 따른 화학물질의 변이원성 구분을 위해, 다른 연구에서는 아직까지 수행된 바 없는 ICR계 마우스의 골수세포를 이용하는 in vivo 소핵시험을 수행하였다. 7주령의 수컷 ICR계 마우스에 동 시험물질의 3가지 농도를 투여하였으며, 경구투여 24시간 후에 도살, 골수세포를 채취하여 슬라이드 표본을 제작하였고, 2,000개의 다염성적혈구 중 소핵을 갖는 다염성 적혈구(MNPCE)를 계수하였다. 모든 투여군에서 cyclohexanone은 골수세포의 증식을 억제하지 않았으며, 소핵을 유발하였다. 이 골수세포를 이용한 소핵시험의 결과로 동 시험물질(cyclohexanone)은 GHS 분류기준에 의거, 변이원성 구분2로 분류하였다.

소 설사병 바이러스 구조단백에 대한 단크론항체 성상에 대한 연구 (Monoclonal antibodies against structural proteins of bovine viral diarrhea virus)

  • 권창희;;우희종
    • 대한수의학회지
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    • 제32권1호
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    • pp.83-90
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    • 1992
  • 소 설사병 바이러스 구조단백에 대한 단크론항체를 작성하여 혈청중화시험, 전기영동, 면역침전반응을 이용하여 분석하였던 바 다음의 결과를 얻었다. 중화능력이 있는 항체의 경우 56K내지 54K의 구조단백에 대응하였다. 그외 중화력을 나타내지 않는 항체는 45K와 36K의 바이러스 항원과 대응하였다. 순수정제된 바이러스의 전기영동 분석결과 12종 이상의 바이러스 단백성분이 구조단백질로서 검출되었으며 중화능력을 나타내는 항체를 이용한 면역침전 결과는 이들의 존재를 뒷받침하였다. 중화단백성분의 세포내 전구물질의 검출은 불가능하였으나 방사선동위원소 부착즉시 세포배지에서 바이러스의 존재를 확인할 수 있었다. Staphylococcus aureus $V_8$효소를 이용한 항원의 부분소화 분석결과 45K와 36K의 바이러스 항원은 서로 상관이 있는 것으로서 입증되었다.

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Intratumoral Administration of Rhenium-188-Labeled Pullulan Acetate Nanoparticles (PAN) in Mice Bearing CT-26 Cancer Cells for Suppression of Tumor Growth

  • Song, Ho-Chun;Na, Kun;Park, Keun-Hong;Shin, Chan-Ho;Bom, Hee-Seung;Kang, Dong-Min;Kim, Sung-Won;Lee, Eun-Seong;Lee, Don-Haeng
    • Journal of Microbiology and Biotechnology
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    • 제16권10호
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    • pp.1491-1498
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    • 2006
  • The feasibility of pullulan acetate nanoparticles (PAN) with ionic strength (IS) sensitivity as a radioisotope carrier to inhibit tumor growth is demonstrated. PAN was radiolabeled with rhenium 188 (Re-188) without any chelating agents. The labeling efficiency of Re-188 into PAN (Re-188PAN) was $49.3{\pm}4.0%$ as determined by TLC. The tumor volumes of mice treated with 0.45 mCi of Re-188-PAN were measured and compared with that of free Re-188 after 5 days of intratumoral injection. For the histological evaluation of apoptotic nuclei of tumor cells, hematoxylin and eosin (H&E), and terminal deoxynucleotidyl transferase biotinylated deoxyuridine triphosphate nick end labeling (TUNEL) staining were performed. The mean tumor volume of the Re-188-PAN-treated group was decreased by 36% after 5 days, whereas that the free Re-188-treated group was decreased by only 15% (P<0.05). The mean number of TUNEL-positive cells in Re-188-PAN-treated tumors at $144.3{\pm}79.9$ cells/section was significantly greater than the control ($26.7{\pm}7.9$ cells/section, P=0.03). The numbers of leukocyte and lymphocyte were decreased in both free Re-188- and Re-188-PAN-treated mice. These results indicated that the intratumoral injection of Re-188-PAN effectively inhibits the tumor growth by prolonging Re-188 retention time in tumor site induced by the IS sensitivity.