• 제목/요약/키워드: DAPI

검색결과 271건 처리시간 0.025초

Investigation of effects of newly synthesized Pt(II) complex against human serum albumin and leukemia cell line of K562

  • Divsalar, Adeleh;Saboury, Ali A.;Ahadi, Leila;Zemanatiyar, Elham;Mansouri-Torshizi, Hassan
    • BMB Reports
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    • 제43권11호
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    • pp.766-771
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    • 2010
  • The biological evaluation of a new synthesized Pt(II)-complex, 2,2'-bipyridin Butylglycinato Pt(II) nitrate, an anti-tumor component, was studied at different temperatures by fluorescence and far UV circular dichroism (CD) spectroscopic methods. Human serum albumin (HSA) and human tumor cell line K562 were as targets. The Pt(II)-complex has a strong ability to quench the intrinsic fluorescence of HSA. Binding and thermodynamic parameters of the interaction were calculated by fluorescence quenching method. Far-UV-CD results showed that Pt(II)-complex induced increasing in content of $\alpha$ helical structure of the protein and stabilized it. The 50% cytotoxic concentration ($Cc_{50}$) of complex was determined using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay at different incubation times. Also, fluorescence staining with DAPI (4,6-diamidino-2-phenylindole) revealed some typical nuclear changes, which are characteristic of apoptosis. Above results suggest that Pt (II) complex is a promising anti-proliferative agent and should execute its biological effects by inducing apoptosis.

Assessment of Characteristics of Biofilm Formed on Autotrophic Denitrification

  • JANG AM;BUM MINSU;KIM SUNGYOUN;AHN YEONGHEE;KIM IN S;BISHOP PAUL L
    • Journal of Microbiology and Biotechnology
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    • 제15권3호
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    • pp.455-460
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    • 2005
  • A pilot-scale sulfur particle autotrophic denitrification (SPAD) process for the treatment of municipal wastewater was operated for 10 months at Shihwa, Korea, and higher than $90\%\;NO^{-}_{3}-N$ removal efficiency was observed. Plate counting showed that the lower part of the denitrifying column reactor had the most autotrophic denitrifiers. The biofilm thickness formed on sulfur particles from the SPAD reactor was approximately $25-30\;{\mu}m$, measured by DAPI (4,6-diamidino-2-phenylindole) staining. The presence of bacteria inside the highly porous sulfur particle was also monitored by SEM observation of the internal surfaces of broken sulfur particles. Biofilm extracellular polymeric substances (EPS) analysis showed that the ratio of carbohydrate to protein decreased with the reactor heights at which biofilm-formed sulfur particles were obtained.

The Distribution of Cytoplasm and Nuclei within the Extra-radical Mycelia in Glomus intraradices, a Species of Arbuscular Mycorrhizal Fungi

  • Lee, Jai-Koo
    • Mycobiology
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    • 제39권2호
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    • pp.79-84
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    • 2011
  • Nuclear distribution within the extra-radical fungal structures and during spore production in the arbuscular mycorrhizae fungus Glomus intraradices was examined using an in vitro monoxenic culture system. A di-compartmental monoxenic culture system was modified using a nitrocellulose membrane and a coverglass slip for detailed observations. Nuclear distribution was observed using the fluorescent DNA binding probes SYBR Green I and DAPI. Both septate and non-septate mycelial regions were observed, but cytoplasmic contents were only found within non-septate mycelia. Nuclear fluorescent staining revealed that the non-septate hyphal region contained nuclei only with cytoplasm, and that nuclear distribution was limited by septa. Swollen hyphal bodies were often associated with septate and empty-looking hyphae. Cytoplasmic contents filled the swollen hyphal body from the non-septate hyphal region following removal of the septa. As a consequence, the swollen body developed into a new spore. These observations provide understanding about the distribution of AM fungal nuclei within extra-radical mycelia and during spore formation. The results suggest a mechanism by which the development of a cytoplasm-containing mycelium is controlled by the formation or removal of septa to efficiently maintain and proliferate essential contents. This mechanism may provide a survival strategy to the fungus.

Combined Effect of Heptaplatin and Ionizing Radiation on Human Squamous Carcinoma Cell Lines

  • Ryu, Mi-Ryeong;Paik, Soon-Young;Chung, Su-Mi
    • Molecules and Cells
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    • 제19권1호
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    • pp.143-148
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    • 2005
  • Heptaplatin, cis-malonato [(4R,5R)-4,5-bis (amino-methyl)-2-isopropyl-1,3-dioxolane] platinum(II) (SKI-2053R, Sunpla) is a new platinum derivative with antitumor activity comparable to cisplatin on various cancer cell lines. Preclinical studies suggest that it is less nephrotoxic than cisplatin. This study was undertaken to examine the combined effect of heptaplatin and ionizing radiation on two established human squamous carcinoma cell lines (NCI-H520, SQ20B). The cytotoxic activity of heptaplatin was concentration-dependent in both cell lines. When low dose heptaplatin was combined with high dose ionizing radiation, there was an additive cytotoxic effect on NCI-H520 cells (P < 0.05), while a moderate dose of heptaplatin and a low dose of ionizing radiation had an additive cytotoxic effect on the growth of SQ20B cells (P < 0.05). FACS analysis and DAPI staining showed that their additive cytotoxic effects were correlated with the induction of apoptosis. Further studies are warranted using heptaplatin and ionizing radiation in squamous cell carcinoma as a substitute for cisplatin.

단삼 추출물이 암세포주에 미치는 세포증식 억제 효과 (Antiproliferative Effect of the Salviae miltiorrhizae Radix Extracts on the Cancer Cell Lines)

  • 양의호;정태산;최창원
    • 대한한의학방제학회지
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    • 제22권2호
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    • pp.35-43
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    • 2014
  • Objectives : The purpose of this study was to identify antiproliferative effects of Salviae miltiorrhizae Radix(SM) extracts against cancer cell lines. Methods : We used 2 kinds of cancer cell lines such as colon cancer cells(HT-29), human oral epitheloid carcinoma cells(KB). MTT assay was performed to examine the efficacy of SM extracts on the cytostaticity of cancer cells in proportion to time and doses. Apoptosis was evaluated by DNA laddering and DAPI nuclei staining. Results : The MTT absorbances against HT-29 and KB of SM extracts were significantly decresed. DNA ladders could be identified in KB of SM extracts. The morphological change were observed and number of cells were decreased by SM extracts. Conclusions : SM extracts is considered to be effective to induce apoptosis and inhibit cancer cell proliferation.

배양 기관지 상피세포(BEAS-2B cells)에서 6가 크롬에 의한 산화적 스트레스 (Induction of Oxidative Stress by Hexavalent Chromium in Human Bronchial Epithelial Cells (BEAS-2B))

  • 박은정;강미선;김대선;박광식
    • Environmental Analysis Health and Toxicology
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    • 제21권4호
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    • pp.357-363
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    • 2006
  • Chromium compounds are widely used in diverse industries including pigment manufacturing, painting, metal plating and leather tanning. With the wide uses of chromium, various adverse effects of the compounds on the environment and human health have been reported. Among them, hexavalent chromium [Cr (VI)], which is a carcinogenic heavy metal, has been widely studies. Epidemiological investigations have shown that respiratory cancers had been found in workers who had been occupationally exposed to Cr (VI). In this study, cell toxicity and induction of reactive oxygen species (ROS) by Cr (VI) (1, 2, 4, $8{\mu}M$) in cultured human bronchial epithelial cells were investigated. Exposure of the cells to Cr (VI) led to cell death, ROS increase, and cytosolic caspase-3 activation. The ROS increase was related with the decreased level of GSH. Chromatin condensation and fragmentation were occurred by Cr (VI) when evaluated by DAPI staining or agarose gel electrophoresis of the extracted DNA. Expression of ROS related genes including glutathione S-transferase, heme oxygenase-1, metallothionein were significantly induced in Cr (VI) treated cells. This result suggests the toxicity in cultured cells by Cr (VI) was expressed through the apoptotic process with ROS induction.

Simple Screening Method for Double-strand DNA Binders Using Hairpin DNA-modified Magnetic Beads

  • Jo, Hun-Ho;Min, Kyoung-In;Song, Kyung-Mi;Ku, Ja-Kang;Han, Min-Su;Ban, Chang-Ill
    • Bulletin of the Korean Chemical Society
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    • 제32권1호
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    • pp.247-250
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    • 2011
  • We designed an effective screening method for double strand DNA (dsDNA) binders using DNA-modified magnetic particles. Hairpin DNA was immobilized on the surface of magnetic particle for a simple screening of dsDNA binding materials in a solution containing various compounds. Through several magnetic separation and incubation processes, four DNA-binding materials, DAPI, 9AA, AQ2A, and DNR, were successfully screened from among five candidates. Efficiency of screening was demonstrated by HPLC analysis using a C2/18 reverse-phase column. In addition, their relative binding strengths were verified by measuring the melting temperature ($T_m$). If hairpin DNA sequence is modified for other uses, this magnetic bead-based approach can be applied as a high-throughput screening method for various functional materials such as anti-cancer drugs.

Phellodendri Cortex Herbal-acupuncture Solution Induced Apoptosis in Human Cervical Cancer Cells, SNU-17

  • Seo, Yong-Seok;Seo, Jung-Chul;Lim, Seong-Chul;Jung, Tae-Young;Han, Sang-Won
    • Journal of Acupuncture Research
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    • 제22권2호
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    • pp.103-110
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    • 2005
  • Phellodendri Cortex (PC) has been used traditionally in Korea for damp heat leukorrhea with thick, yellow, discharge, foul-smelling diarrhea or dysentery. We investigated whether the Phellodendri Cortex Herbal-acupuncture Solution (PCHS) induced cell-death on SNU-17, human cervical cancer cell. 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was performed to find out the cytotoxicity of PCHS. The cell death was identified as apoptosis from the results of 4, 6-diamidineo-2-phenylindole (DAPI) staining, terminal deoxy-nucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay. The expression of proapototic gene, Bax, was increased and the expression of apoptotic gene, Caspase-3, was also increased. Considering the above results, PCHS could induce the apoptosis on SNU-17, human cervical cancer cell, via Bax-related Caspase-3 activation. And it might provide the experimental data for the clinical use of Phellodendri Cortex on cervical cancer.

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The Bacterial Community of Southern Lake Baikal in Winter

  • Ahn, Tae-Seok;Hong, Sung-Hee;Kim, Dong-Joo;Suck, Jung-Hyun;Valentin V. Drucker
    • Journal of Microbiology
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    • 제37권1호
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    • pp.10-13
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    • 1999
  • The bacterial abundance, proportion of respiring bacteria, and bacterial community of southern Lake Baikal were analyzed at 1 m and 400m depths during winter. The total bacterial numbers were 5.1${\times}$105 cells ml-1 at 1 m and 2.5${\times}$105 cells ml-1 at 400 m depth, which are about half and quarter of the numbers of other lakes. The proportion of respiring bacteria was as low as 2.5% at 1 m and 1.4% at 400 m depth. Considering the amount of organic carbon which need to be degraded and low proportion of respiring bacteria, the bacteria could be assumed to have high activities. The EUB/DAPI ratios were 77 and 89% at 1 m and 400 m depths, respectively. Of the bacterial community, the other group was dominant at both depths, and gamma group of protebacteria followed next. But the beta group of proteobacteria and Cytophaga-Flavobacterium groups occupied very small proportions.

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KINETICS OF AUTOTROPHIC DENITRIFICATION FOR THE BIOFILM FORMED ON SULFUR PARTICLES : Evaluation of Molecular Technique on Monitoring Biomass Growth

  • Kim, Sung-Youn;Jang, Am;Kim, I-Tae;Kim, Kwang-Soo;Kim, In-S.
    • Environmental Engineering Research
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    • 제10권6호
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    • pp.283-293
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    • 2005
  • Characteristics of sulfur-based autotrophic denitrification in a semi-continuous type reactor and the kinetic parameters were studied. Enriched autotrophic denitrifying culture was used for the reactor operation. Biomass growth on sulfur particles and in the liquid medium was monitored using the DAPI staining method. From the result of ion concentration changes and the biomass growth, maximum specific growth rate, ${\mu}_{max}$, and the half velocity constant, $K_M$, were estimated as $0.61\;d^{-1}$ and 3.66 mg/L, respectively. Growth yield coefficient, Y values for electron acceptor and donor were found as 0.49 gVSS/g N and 0.16 gVSS/g S. The biomass showed specific denitrification rate, ranging 0.86-1.13 gN/g VSS-d. A half-order equation was found to best simulate the denitrification process in the packed bed reactor operated in the semi-continuous mode.