• Title, Summary, Keyword: Hoechst 33258

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Genome Detection Using Hoechst 33258 Groove Binder (Hoechst 33258 Groove Binder를 이용한 DNA칩)

  • Choi, Yong-Sung;Lee, Kyung-Sup
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • pp.372-373
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    • 2006
  • In this paper, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5 end were immobilized on the gold electrodes by DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes.

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Electrochemical Gene Detection Using Hoechat Groove Binder (Hoechst groove binder를 이용한 유전자의 전기화학적 검출)

  • Choi, Yong-Sung;Lee, Woo-Ki;Lee, Kyung-Sup
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • pp.65-70
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    • 2006
  • In this study, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5 end were immobilized on the gold electrodes by DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes.

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Simple Classification of Male Mouse Germ Cells using Hoechst 33258 Staining (Hoechst 33258 Staining을 이용한 웅성 생쥐 성세포의 간편 분류)

  • Kim, Kyoung Guk;Park, Young Sik
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.213-218
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    • 2015
  • In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

Electrochemical Gene Detection Using Microelectrode Array on a DNA Chip

  • Park, Yong-Sung;Kwon, Young-Soo;Park, Dae-Hee
    • KIEE International Transactions on Electrophysics and Applications
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    • v.4C no.4
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    • pp.145-148
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    • 2004
  • In this study, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5' end were immobilized on the gold electrodes by a DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 and concentrated at the electrode surface through association with the formed hybrid. This suggested that this DNA chip could recognize the sequence specific genes.

Eletrochemical Detection of Gene using Microelectrode-array DNA Chip (미소전극어레이형 DNA칩을 이용한 유전자의 전기화학적 검출)

  • ;;Eiichi Tamiya
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.17 no.7
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    • pp.729-737
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    • 2004
  • In this paper, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5 end were immobilized on the gold electrodes by DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes.

Genomic Detection using Electrochemical Method (전기화학적 방법에 의한 유전자의 검출)

  • Choi, Yong-Sung;Lee, Kyung-Sup;Park, Dae-Hee
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.18 no.6
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    • pp.560-570
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    • 2005
  • In this paper, a microelectrode away DNA chip was fabricated on glass slide using photolithography technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5' end were immobilized on the gold electrodes by DNA arrayer utilizing the affinity between gold and sulfu. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Cyclic voltammetry in 5mA ferricyanide/ferrocyanide solution at 100 mV/s confirmed the immobilization of probe DNA on the gold electrodes. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic system.

Electrochemical Detection of Genes Using Microeledtrode Array DNA Chip (미소전극어에이형 DNA칩을 이용한 유전자의 전기화학적 검출)

  • Choi, Yong-Sung;Park, Dae-Hee
    • Proceedings of the KIEE Conference
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    • pp.2125-2127
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    • 2004
  • In this paper, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5 end were immobilized on the gold electrodes by DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes.

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Electrochemical Detection of Single Nucleotide Polymorphism (SNP) Using Microelectrode Array on a DNA Chip (미소전극어레이형 DNA칩을 이용한 유전자다형의 전기화학적 검출)

  • 최용성;권영수;박대희
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.53 no.5
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    • pp.286-292
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    • 2004
  • In this study, an integrated microelectrode array was fabricated on glass slide using microfabrication technology. Probe DNAs consisting of mercaptohexyl moiety at their 5-end were spotted on the gold electrode using micropipette or DNA arrayer utilizing the affinity between gold and sulfur. Cyclic voltammetry in 5mM ferricyanide/ferrocyanide solution at 100 ㎷/s confirmed the immobilization of probe DNA on the gold electrodes. When several DNAs were detected electrochemically, there was a difference between target DNA and control DNA in the anodic peak current values. It was derived from specific binding of Hoechst 33258 to the double stranded DNA due to hybridization of target DNA. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic System.

Applications of capacitation status for litter size enhancement in various pig breeds

  • Kwon, Woo-Sung;Shin, Dong-Ha;Ryu, Do-Yeal;Khatun, Amena;Rahman, Md Saidur;Pang, Myung-Geol
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.6
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    • pp.842-850
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    • 2018
  • Objective: Several studies have reported the development of new molecular methods for the prognosis and diagnosis of male fertility based on biomarkers aimed at overcoming the limitations of conventional male fertility analysis tools. However, further studies are needed for the field application of these methods. Therefore, alternative methods based on existing semen analysis methods are required to improve production efficiency in the animal industry. Methods: we examined the possibility of improving litter size in various pig breeds using combined Hoechst 33258/chlortetracycline fluorescence (H33258/CTC) staining. The correlation between field fertility and capacitation status by combined H33258/CTC staining in different ejaculates spermatozoa (n = 3) from an individual boar (20 Landrace, 20 Yorkshire, and 20 Duroc) was evaluated as well as overall accuracy. Results: The acrosome reacted (AR) pattern after capacitation (%) was positively correlated with the litter size of Landrace, Yorkshire, and Duroc pigs and the overall accuracy was 75%, 75%, and 70% in Landrace, Yorkshire, and Duroc pigs, respectively. The difference (${\Delta}$) in AR pattern before and after capacitation was positively correlated with the litter size of Landrace, Yorkshire, and Duroc pigs and the overall accuracy was 80%, 65%, and 55% in Landrace, Yorkshire, and Duroc pigs, respectively. However, the difference (${\Delta}$) in capacitated (B) pattern before and after capacitation was negatively correlated with the litter size of Landrace pigs and the overall accuracy was 75%. Moreover, average litter size was significantly altered according to different combined H33258/CTC staining parameters. Conclusion: These results show that combined H33258/CTC staining may be used to predict male fertility in various breeds. However, the selection of specific efficiency combined H33258/CTC staining parameters requires further consideration. Taken together, these findings suggest that combined H33258/CTC staining may constitute an alternative method for predicting male fertility until such time as fertility-related biomarkers are further validated.

Cytotoxicity of COX-2 Inhibitor (Nimesulide) in Non-small Cell Lung Cancer Cell Line (비소세포폐암 세포주에서 COX-2억제제(Nimesulide)의 세포독성)

  • Park Chan Beom;Jeon Hyun Woo;Jin Ung;Cho Kyu Do;Kim Chi Kyung;Wang Young-Pil
    • Journal of Chest Surgery
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    • v.38 no.4
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    • pp.263-270
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    • 2005
  • In recent years, a combination of two demographic phenomena, an increased number of older people in the population and an increase in the incidence of lung cancer with age, has made it mandatory to develop therapeutic modalities with less toxicity for the treatment of inoperable elderly patients with lung cancer. Therefore, we investigated the correlation between COX-2 expression and cytotoxicity of Nimesulide, a specific COX-2 inhibitor. Material and Method: Immunohistochemical staining of COX-2 was performed. After exposure of Nimesulide, XTT analysis, FACS analysis and Hoechst staining were carried out. Result: COX-2 protein was expressed in non-treated A549 cells strongly, but not in H1299. Cytotoxicity of Nimesulide against A549 cell and H1299 cell were similar and $IC_{50}$ of Nimesulide in both cell lines were $70.9{\mu}M$ in A549 cell line and $56.5{\mu}M$ in H1299 cell line respectively. FACS analysis showed $G_0/G_1$ arrest in both cell lines and the S phase cell fraction was decreased. Morphologic assessment of apoptosis by Hoechst 33258 staining, many apoptotic cells were detected in both cell lines. Conclusion: Selective COX-2 inhibitor, Nimesulide, can inhibit the proliferation of non-small cell lung cancer cell lines in vitro. Inhibitory effect of Nimesulide are induction of apoptosis and $G_0/G_1$ arrest. There is no correlation between COX-2 expression and cytotoxicity of Nimesulide, a specific COX-2 inhibitor. Therefore, highly selective COX-2 inhibitors such as Nimesulide can be expected to lead to even greater efficacy of their use as adjuncts to various anticancer angents and radiation therapy for the treatment of high-risk patients.