• 제목/요약/키워드: In vivo detection

검색결과 184건 처리시간 0.027초

Detection of DNA Damage in Carp Using Single-Cell Gel Electrophoresis Assay for Genotoxicity Monitoring

  • Jin, Hai-Hong;Lee, Jae-Hyung;Hyun, Chang-Kee
    • Journal of Microbiology and Biotechnology
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    • 제14권2호
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    • pp.268-275
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    • 2004
  • To investigate the potential application of the single-cell gel electrophoresis (SCGE) assay to carp as an aquatic pollution monitoring technique, gill, liver, and blood cells were isolated from carp exposed to a direct-acting mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), or indirect mutagen, $benzo[\alpha]pyrene$ $(B[\alpha]P)$, then the DNA strand breakage was analyzed using the assay. Based on testing 5 different cell isolation methods and 6 electrophoretic conditions, the optimized assay conditions were found to be cell isolation by filter pressing and electrophoresis at a lower voltage and longer running time (at 0.4 V/cm for 40 min). In preliminary experiments, gill and liver cells isolated from carp exposed to MNNG in vitro exhibited DNA damage signals even with 0.5 ppb exposure, which is a much higher dose than previously reported. In the gill cells isolated from carp exposed to 0.01-0.5 ppm MNNG in vivo, significant dose-and time-dependent increases were observed in the tail for 4 days. As such, the linear correlation between the relative damage index (RDI) values and time for each dose based on the initial 48-h exposure appeared to provide effective criteria for the genotoxicity monitoring of direct-acting mutagenic pollution. In contrast, the in vivo exposure of carp to 0.25-1.0 ppm of $B[\alpha]P$ for 7 days resulted in dose-and time-dependent responses in the liver cells, in which 24-h delayed responses for metabolizing activation and gradual repair after 48 h were also observed. Thus, the negative-sloped linear correlation between the RDI and time at each dose based on the initial 48 h appeared to provide more effective criteria for the genotoxicity monitoring of indirect mutagenic pollution.

Comet Assay to Detect the DNA Breakages in the Tissue of the Purple Clam ( Saxidomus purpuratus) and the Blood of the Olive Flounder (Paralichthys olivaceus) Exposed to 5 PAHs

  • Lee, Taek-Kyun;Kim, So-Jung;Park, Eun-Seok;Rora Oh;Yun, Hee-Young;Man Chang
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2003년도 추계국제학술대회
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    • pp.159-159
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    • 2003
  • Comet assay is a potential monitoring tool because DNA strand breakage may be produced by a wide range of agents. The comet assay, also called the single-cell gell electrophoresis (SCGE) assay, is rapid and sensitive method for the detection of DNA damage in cells. This study was performed for the identification of DNA damage in the cells from flounders and clams exposed to PAHs. As a control experiments, flounder and clam cells were exposed to $H_2O$$_2$. The cells exposed to $H_2O$$_2$ were displayed a typical nuclei movement DNA damage of cells were significantly increased when the isolated cells from the blood of flounders and the tissue of clams were in vitro exposed to the different concentrations (5, 10, 50, 100 ppb) of five kinds of PAHs (benzo[a]pyrene, pyrene, fluoranthene, anthrancene, and phenanthrene). For the in vivo test, flounders and clams were exposed to the different concentrations of BaP for 4 days. The results showed that DNA strand breakage was effected by the concentration of BaP and the duration of exposure. In high concentration of BaP, the mean tail lengths of nuclei was longer than it In low concentration, while the mean size of head DNA decreased. In this research, both in vitro and in vivo genotoxicity of PAHs could be biomonitored by the comet assay. Especially, clams and flounders seem to be useful as materials for monitoring genotoxic damage by comet assay.

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가변편광과 광학배율을 기반으로 한 피부 확대경을 이용한 피부주름 측정 (Skin Wrinkle Detection Using Dermatologic Magnifier Based on Variable Polarization and Optical Magnification)

  • 배영우;정병조
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2006년 학술대회 논문집 정보 및 제어부문
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    • pp.190-192
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    • 2006
  • The clinical characteristics of photodamaged skin, such as coarse and fine wrinkle are not accurately evaluable with previous methods. Public awareness for wrinkle treatment and prevention which rely on proper assessment and evaluation of the underlying skin changes has been increased as the population ages. In this paper, we suggest an in-vivo method and instrument that allow us to acquire a wrinkle-enhanced image to improve the accuracy of quantitative and qualitative analysis for skin wrinkle. The method used involved white LED illumination and photography through polarizing filters. Finally, the polarized light photography yields an accurate and evaluable parameter of photodamaged skin, especially fine and coarse wrinkle.

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Fabrication of Chitosan-gold Nanocomposites Combined with Optical Fiber as SERS Substrates to Detect Dopamine Molecules

  • Lim, Jae-Wook;Kang, Ik-Joong
    • Bulletin of the Korean Chemical Society
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    • 제35권1호
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    • pp.25-29
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    • 2014
  • This research was aimed to fabricate an optical fiber-based SERS substrate which can detect dopamine neurotransmitters. Chitosan nanoparticles (NPs) were firstly anchored on the surface of optical fiber, and then gold layer was subsequently deposited on the anchored chitosan NPs via electroless plating method. Finally, chitosan-gold nanocomposites combined with optical fiber reacted with dopamine molecules of 100-1500 mg/day which is a standard daily dose for Parkinson's disease patients. The amplified Raman signal at $1348cm^{-1}$ obtained from optical fiber-based SERS substrate was plotted versus dopamine concentrations (1-10 mM), demonstrating an approximate linearity of Y = 303.03X + 2385.8 ($R^2$ = 0.97) with narrow margin errors. The optical fiber-based Raman system can be potentially applicable to in-vitro (or in-vivo) detection of probe molecules.

Murine Heart Wall Imaging with Optical Coherence Tomography

  • Kim Jee-Hyun;Lee Byeong-Ha
    • Journal of the Optical Society of Korea
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    • 제10권1호
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    • pp.42-47
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    • 2006
  • M-mode imaging of the in vivo murine myocardium using optical coherence tomography (OCT) is described. Application of conventional techniques (e.g. MRI, Ultrasound imaging) for imaging the murine myocardium is problematic because the wall thickness is less than 1.5 mm (20 g mouse), and the heart rate can be as high as six hundred beats per minute. To acquire a real-time image of the murine myocardium, OCT can provide sufficient spatial resolution ($10{\mu}m$) and imaging speed (1000 A-scans/s). Strong light scattering by blood in the heart causes significant light attenuation, which makes delineation of the endocardium-chamber boundary problematic. To measure the thickness change of the myocardium during one heart beat cycle, a myocardium edge detection algorithm is developed and demonstrated.

Characterization of Norepinephrine Release in Rat Posterior Hypothalamus Using in vivo Brain Microdialysis

  • Sung, Ki-Wug;Kim, Seong-Yun;Kim, Ok-Nyu;Lee, Sang-Bok
    • The Korean Journal of Physiology and Pharmacology
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    • 제6권1호
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    • pp.9-14
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    • 2002
  • In the present study, we used the microdialysis technique combined with high performance liquid chromatography (HPLC) and electrochemical detection to measure the extracellular levels of norepinephrine (NE) in the posterior hypothalamus in vivo, and to examine the effects of various drugs, affecting central noradrenergic transmission, on the extracellular concentration of NE in the posterior hypothalamus. Microdialysis probes were implanted stereotaxically into the posterior hypothalamus (coordinates: posterior 4.3 mm, lateral 0.5 mm, ventral 8 mm, relative to bregma and the brain surface, respectively) of rats, and dialysate collection began 2 hr after the implantation. The baseline level of monoamines in the dialysates were determined to be: NE $0.17{\pm}0.01,$ 3,4-dihydroxyphenylacetic acid (DOPAC) $0.94{\pm}0.07,$ homovanillic acid (HVA) $0.57{\pm}0.05$ pmol/sample (n=8). When the posterior hypothalamus was perfused with 90 mM potassium, maximum 555% increase of NE output was observed. Concomitantly, this treatment significantly decreased the output of DOPAC and HVA by 35% and 28%, respectively. Local application of imipramine $(50\;{\mu}M)$ enhanced the level of NE in the posterior hypothalamus (maximum 200%) compared to preperfusion control values. But, DOPAC and HVA outputs remained unchanged. Pargyline, an irreversible monoamine oxidase inhibitor, i.p. administered at a dose of 75 mg/kg, increased NE output (maximum 165%), while decreased DOPAC and HVA outputs (maximum 13 and 12%, respectively). These results indicate that NE in dialysate from the rat posterior hypothalamus were neuronal origin, and that manipulations which profoundly affected the levels of extracellular neurotransmitter had also effects on metabolite levels.

자초 뿌리의 함유성분 및 아토피피부염 관련 연구현황 (Progress on Phytochemical and Atopic Dermatitis-related Study of the Root of Lithospermum erythrorhizon)

  • 주지훈;조현환;이용섭
    • 생약학회지
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    • 제41권2호
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    • pp.73-88
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    • 2010
  • Traditionally, the root of Lithospermum erythrorhizon Sieb. et Zucc(L.E) has been used as efficacious therapy for inflammation, burns, frostbite and skin ailments (e.g eczema and psoriasis). It contains isohexenylnaphthoquinone derivatives (shikonin and its esters) and furylhydroquinones (shikonofurans) in lipophilic fractions and caffeic acid oligomers (rosmarinic acid, lithospermic acid B) in polar fractions. Recently, new preparative isolation and analysis procedures of shikonin along with its oligomers from the extract of L. erythrorhizon by the combination of high-speed counter-current chromatography with high-performance liquid chromatography-diode array detection have also been introduced. Although there have been many reports on the wound healing, antiinflammatory, and anticancer effects, the research on the effects of anti-atopic dermatitis of the root of L. erythrorhizon were relatively scarce. However, in recent years, new information gathered from research efforts, on the anti-atopic dermatitis properties of the extract or constituents of L. erythrorhizon has been accumulated. In this paper, the findings and advance on the in vitro and in vivo activities of L. erythrorhizon and its constituents especially focused on antiinflammatory and anti-atopic dermatitis effects are summarized. The phytochemical constituents of L. erythrorhizon or its tissue cultures are also presented. Although there are few to verify or refute its activity in human, one result of clinical study of the extract of L. erythrorhizon on the atopic dermatitis patients was introduced to assess the possibility of its clinical use. The reported mechanisms of action and in vivo pharmacological studies in different animal models for the various types of extracts or constituents of L. erythrorhizon are supportive of its therapeutic potential or dietary supplement, however, more evidence from clinically relevant models, as well as systemic studies on the active constituents or the various types of standardized extracts at the cellular and molecular level, are required.

LAMP 방법에 의한 소 수정란의 성 판별과 Biopsy에 따른 수정란의 체외발달 (Sex Detection and In Vitro Development of Biopsied Bovine Embryo for LAMP Based Embryo Sexing)

  • 조상래;최선호;김현종;한만희;최창용;정연길;손동주
    • 한국수정란이식학회지
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    • 제20권2호
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    • pp.169-176
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    • 2005
  • 수정란의 성 판별은 유전적으로 우수한 유전형질을 보유하고 있는 소의 수정란을 성 판별하므로서 희망하는 성의 송아지를 생산할 수 있으며, 부가가치가 높은 수정란을 확보할 수 있는 기술이다. 수정란의 손상을 최소화하면서 할구를 biopsy하는 기술을 개발하고, 간단하고 빠른 시간에 성 판별이 가능한 Loop-mediated isothermal amplification방법으로 수정란을 성 판별을 실시한 결과는 다음과 같다. 1. 한우 체내 수정란의 성비는 암컷이 $56.5\%$, 수컷이 $43.5\%$였고, 체외 수정란은 암컷이 $49.2\%$, 수컷이 $33.9\%$였다. 그리고, 젖소 체외 수정란의 성비는 암컷이 $29.2\%$, 수컷이 $70.8\%$를 나타내어 한우 체내 및 체외 수정란보다 젖소 체외수정란의 수컷비율이 유의적으로 높게 나타내었다(p<0.05). 또한, 한우 체외 수정란에서 성 판별이 불가능한 것이 $16.9\%$를 나타내어 한우 체내 수정란 및 젖소 체외 수정란과는 유의적인 차이를 나타내었다.(P<0.05). 2. Biopsy한 체내 수정란의 체외 발달율은 $100\%$였으나 체외 수정란에서는 정상적으로 발달하지 못하고 퇴화된 수정란이 $13.2\%$로 체내 수정란보다 유의적으로 높은 결과를 나타내었다.(P<0.05). 3. Punching 방법으로 수정란의 biopsy 후 정상적으로 발달하지 못하고 퇴화된 수정란은 체내 및 체외 수정란에서는 없었으나 biopsy 방법으로 biopsy한 수정란은 체내 및 체외 수정란에서 각각 $16.7\%$$22.6\%$를 나타내어 Punching 방법보다 유의적으로 높은 결과를 나타내었다(P<0.05). 이상의 결과로 보아 한우 체내 수정란은 LAMP방법을 이용하여 간단하고 신속하게 성 판별이 가능하며, 수정란의 biopsy는 punching 방법이 수정란에 손상을 적게 주는 것으로 사료된다.

혈류 방향을 구별하는 연속 초음파 도플러 장치에 관한 연구 (A study on the development of CW(Continuous-Wave) Doppler system for measuring bi-directional blood flow information.)

  • 강충신;김영길
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 1987년도 전기.전자공학 학술대회 논문집(II)
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    • pp.1239-1242
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    • 1987
  • With the convention CW Doppler velocity meter, bi-directional velocities cannot be separated. The new CW Doppler system usee quadrature detection and phase rotation to Produce simultaneous independent audio and velocity signals for forward and reverse blood flow direction, is fabricated. Specially, this system shows that phase rotation method for flow direction separation provides easy and satisfactory feature. From in vivo blood flow measurement, can easily differentiate typical artery flow from vein flow. and measure both velocity characteristics qualitatively.

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