• Title/Summary/Keyword: Bioluminescence

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MicroSPECT and MicroPET Imaging of Small Animals for Drug Development

  • Jang, Beom-Su
    • Toxicological Research
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    • v.29 no.1
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    • pp.1-6
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    • 2013
  • The process of drug discovery and development requires substantial resources and time. The drug industry has tried to reduce costs by conducting appropriate animal studies together with molecular biological and genetic analyses. Basic science research has been limited to in vitro studies of cellular processes and ex vivo tissue examination using suitable animal models of disease. However, in the past two decades new technologies have been developed that permit the imaging of live animals using radiotracer emission, X-rays, magnetic resonance signals, fluorescence, and bioluminescence. The main objective of this review is to provide an overview of small animal molecular imaging, with a focus on nuclear imaging (single photon emission computed tomography and positron emission tomography). These technologies permit visualization of toxicodynamics as well as toxicity to specific organs by directly monitoring drug accumulation and assessing physiological and/or molecular alterations. Nuclear imaging technology has great potential for improving the efficiency of the drug development process.

Monitoring of Bioluminescent Lactobacillus plantarum in a Complex Food Matrix

  • Moon, Gi-Seong;Narbad, Arjan
    • Food Science of Animal Resources
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    • v.37 no.1
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    • pp.147-152
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    • 2017
  • A bioluminescent Lactobacillus plantarum (pLuc2) strain was constructed. The luminescent signal started to increase during the early exponential phase and reached its maximum in the mid-exponential phase in a batch culture of the strain. The signal detection sensitivity of the strain was the highest in PBS (phosphate buffered saline), followed by milk and MRS broth, indicating that the sensitivity was influenced by the matrix effect. The strain was used in millet seed fermentation which has a complex matrix and native lactic acid bacteria (LAB). The luminescent signal was gradually increased until 9 h during fermentation and abolished at 24 h, indicating that the strain could be specifically tracked in the complex matrix and microflora. Therefore, the bioluminescent labeling system can be used for monitoring LAB in food and dairy sciences and industries.

Characterization of gltA::luxCDABE Fusion in Escherichia coli as a Toxicity Biosensor

  • Ahn, Joo-Myung;Kim, Byoung-Chan;Gu, Man-Bock
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.6
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    • pp.516-521
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    • 2006
  • The use of gltA gene, as a new biomarker for environmental stress biomonitoring, was investigated because of its key position as the first enzyme of the tricarboxylic acid (TCA) cycle. A recombinant bioluminescent Escherichia coli strain, EBJM2, was constructed using a plasmid carrying the citrate synthase (gltA) promoter transcribing the Photorhabdus luminescens IuxCDABE genes (gltA::luxCDABE). The responses from this strain were studied with five different classes of toxicants: DNA damage chemicals, phenolics, oxidative-stress chemicals, PAHs, and organic solvents. EBJM2 responded strongly to DNA damage chemicals, such as mitomycin C (MMC) and methyl-nitro-nitrosoguanidine (MNNG) and nalidixic acid with the strongest responses. In contrast, tests with several compounds from the other four classes of toxicants gave no significant response. Therefore, EBJM2 was found to be sensitive to DNA damage chemicals.

Fluorescent and Luminescent Proteins Derived from Marine Organisms: Functions and Applications

  • Sehyeok, Im;Jisub, Hwang;Hackwon, Do;Bo-Mi, Kim;Sung Gu, Lee;Jun Hyuck, Lee
    • Journal of Marine Life Science
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    • v.7 no.2
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    • pp.74-85
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    • 2022
  • Organisms constituting a large proportion of marine ecosystems, ranging from bacteria to fish, exhibit fluorescence and bioluminescence. A variety of marine organisms utilize these biochemically generated light sources for feeding, reproduction, communication, and defense. Since the discovery of green fluorescent protein and the luciferin-luciferase system more than a century ago, numerous studies have been conducted to characterize their function and regulatory mechanism. The unique properties of fluorescent and bioluminescent proteins offer great potential for their use in a broad range of applications. This short review briefly describes the functions and characteristics of fluorescent and bioluminescent proteins, in addition to summarizing the recent status of their applications.

QSAR Modeling of Toxicant Concentrations(EC50) on the Use of Bioluminescence Intensity of CMC Immobilized Photobacterium Phosphoreum (CMC 고정화 Photobacterium phosphoreum 의 생체발광량을 이용한 독성농도(EC50)의 QSAR 모델)

  • 이용제;허문석;이우창;전억한
    • KSBB Journal
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    • v.15 no.3
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    • pp.299-306
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    • 2000
  • Concern for the effects of toxic chemicals on the environment leads the search for better bioassay test organisms and test procedures. Photobacterium phosphoreum was used successfully as a test organism and the luminometer detection technique was an effective and simple method for determining the concentration of toxic chemicals. With EC50 a total of 14 chlorine substituted phenols benzenes and ethanes were used for the experiments. The test results showed that the toxicity to P. phosphoreum increased in the order of phenol > benzene > ethane and the toxicity also increased with the number of chlorine substitution. Quantitative structure activity relationship (QSARO) model can be used to predict EC50 to save time and endeavor. Correlation was well established with the QSAR parameters such as log P, log S and solvatochromic parameter(Vi/100 $\pi$, ${\beta}$m and am). The QSAR modeling was used with multi-regression analysis and mono-regression analysis. These analyses resulted in the following QSAR : $log EC_{50} =2.48 + 0.914 log S(n=9 R2=85.5% RE=0.378) log EC_{50}=0.35 - 4.48 Vi/100 + 2.84 \pi^* +9.46{\beta}m-4.48am (n =14 R2=98.2% RE=0.012) log EC_{50} =2.64 -1.66 log P(n=5, R2=98.8% RE=0.16) log EC_{50}=3.44 -1.09 log P(n=9 R2= 80.8% Re=0.207)$

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Bioassessment of Heavy Metals, Nanoparticles, and Soils Contaminated with Metals using Various Bioassays (다양한 독성법을 이용한 중금속, 나노입자 및 금속오염 토양 평가)

  • Kong, In Chul;Shi, Yu Tal;Lee, Min Kyung;Kang, Il Mo
    • Economic and Environmental Geology
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    • v.48 no.3
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    • pp.261-271
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    • 2015
  • Toxicity results of metals, nanoparticles (NPs), and soils contaminated with metals were introduced on this review. Following methods were used: seed germination, bioluminescence, enzyme activity, and mutation. In general, different sensitivities were observed, depending on types of bioassays and pollutants. Among tested seeds, sensitivities of Lactucus and Raphanus were greater than others. Of single metal exposure, effect by As(III) was greater than others, and high revertant mutation ratio (5.1) was observed at 1 mg/L arsenite, indicating high mutagenicity. No general pattern was observed on the effect of metal mixture, but synergistic effect was observed with seeds. In case of soils, no correlation was observed between total metal contents and toxicity. Toxicity of NPs was observed as follows: CuO > ZnO > NiO > $TiO_2$, $Fe_2O_3$, $Co_3O_4$. Especially, no considerable effects were observed by $TiO_2$, $Fe_2O_3$, and $Co_3O_4$ under tested concentration (max. 1,000 mg/L). The evaluation results of interactive toxic effects using various bioassays may comprise a useful tool for the bioassessment of various environmental pollutants.

Verification of the Suitability of the ATP Luminometer as the Monitoring Tool for Surface Hygiene in Foodservices (급식소 식품접촉표면 위생 모니터링 도구로서의 ATP Luminometer 적합성 확인)

  • Kim, Yang-Sook;Moon, Hey-Kyung;Kang, Seong-Il;Nam, Eun-Jeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.11
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    • pp.1719-1723
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    • 2010
  • The purpose of this study was to verify feasibility of using a ATP Luminometer, real-time hygiene monitoring tool for food contact surfaces in foodservices. For this, 54 cutting boards, 70 knives, 21 rubber gloves in 4 institutional foodservices were studied. ATP (RLU: relatively light unit) values by ATP Luminometer were compared with APC (CFU: colony forming unit) of swabbing culture method using aerobic count plates of 3M petrifilm. ATP ranged from 0 RLU/$cm^2$ to 64693 RLU/$cm^2$ on knives, from 0.1 RLU/$cm^2$ to 6743.6 RLU/$cm^2$ on cutting boards and from 31 RLU/$cm^2$ to 465635 RLU/$cm^2$ on the rubber gloves. APC ranged from 0 CFU/$cm^2$ to 166667 CFU/$cm^2$ on knives, from 0 CFU/$cm^2$ to 1000 CFU/$cm^2$ on cutting boards and from 0 CFU/$cm^2$ to 730000 CFU/$cm^2$ on the rubber gloves. To express the degree of association between ATP and APC, a linear regression was performed. There were significant positive correlations found between log RLU and log CFU on the knives (r=0.84, p<0.001), the cutting boards (r=0.79, p<0.001), the rubber gloves (r=0.78, p<0.001). Results of this study showed the possibility that ATP bioluminescence technique can be used as the monitoring tool for surface hygiene in foodservices.

Bioluminescence Imaging of Chondrocytes in Rabbits by Intraarticular Injection of D-Luciferin (토끼에서 D-luciferin의 관절강 주입에 의한 연골세포의 자연발광 영상)

  • Moon, Sung-Min;Min, Jung-Joon;Oh, Suk-Jung;Kang, Han-Saem;Kim, Young-Ho;Kim, Sung-Mi;Kim, Kwang-Yoon;Bom, Hee-Seung
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.1
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    • pp.54-58
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    • 2007
  • Purpose: Luciferase is one of the most commonly used reporter enzymes in the field of in vivo optical imaging. D-luciferin, the substrate for firefly luciferase has very high cost that allows this kind of experiment limited to small animals such as mice and rats. In this current study, we validated local injection of D-luciferin in the articular capsule for bioluminescence imaging in rabbits. Materials and Methods: Chondrocytes were cultured and infected by replication-defective adenoviral vector encoding firefly luciferase (Fluc). Chondrocytes expressing Fluc were injected or implanted in the left knee joint. The rabbits underwent optical imaging studies after local injection of D-luciferin at 1, 5, 7, 9 days after cellular administration. We sought whether optimal imaging signals was could be by a cooled CCD camera after local injection of D-luciferin. Results: Imaging signal was not observed from the left knee joint after intraperitoneal injection of D-luciferin (15 mg/kg), whereas it was observed after intraarticular injection. Photon intensity from the left knee joint of rabbits was compared between cell injected and implanted groups after intraarticular injection of D-luciferin. During the period of imaging studies, photon intensity of the cell implanted group was 5-10 times higher than that of the cell injected group. Conclusion: We successfully imaged chondrocytes expressing Fluc after intraarticular injection of D-luciferin. This technique may be further applied to develop new drugs for knee joint disease.

Development of Optical Molecular Imaging System for the Acquisition of Bioluminescence Signals from Small Animals (소동물 발광영상 측정을 위한 광학분자영상기기의 개발)

  • Lee, Byeong-Il;Kim, Hyeon-Sik;Jeong, Hye-Jin;Lee, Hyung-Jae;Moon, Seung-Min;Kwon, Seung-Young;Choi, Eun-Seo;Jeong, Shin-Young;Bom, Hee-Seung;Min, Jung-Joon
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.4
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    • pp.344-351
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    • 2009
  • Purpose: Optical imaging is providing great advance and improvement in genetic and molecular imaging of animals and humans. Optical imaging system consists of optical imaging devices, which carry out major function for monitoring, tracing, and imaging in most of molecular in-vivo researches. In bio-luminescent imaging, small animals containing luciferase gene locally irradiate light, and emitted photons transmitted through skin of the small animals are imaged by using a high sensitive charged coupled device (CCD) camera. In this paper, we introduced optical imaging system for the image acquisition of bio-luminescent signals emitted from small animals. Materials and Methods: In the system, Nikon lens and four LED light sources were mounted at the inside of a dark box. A cooled CCD camera equipped with a control module was used. Results: We tested the performance of the optical imaging system using effendorf tube and light emitting bacteria which injected intravenously into CT26 tumor bearing nude mouse. The performance of implemented optical imaging system for bio-luminescence imaging was demonstrated and the feasibility of the system in small animal imaging application was proved. Conclusion: We anticipate this system could be a useful tool for the molecular imaging of small animals adaptable for various experimental conditions in future.

Development of Quantification Method for Bioluminescence Imaging (발광영상에 대한 정량화 방법 개발)

  • Kim, Hyeon-Sik;Choi, Eun-Seo;Tak, Yoon-O;Choi, Heung-Kook;Lee, Ju-Young;Min, Jung-Joon;Lee, Byeong-Il
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.451-458
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    • 2009
  • Purpose: Optical molecular luminescence imaging is widely used for detection and imaging of bio-photons emitted by luminescent luciferase activation. The measured photons in this method provide the degree of molecular alteration or cell numbers with the advantage of high signal-to-noise ratio. To extract useful information from the measured results, the analysis based on a proper quantification method is necessary. In this research, we propose a quantification method presenting linear response of measured light signal to measurement time. Materials and Methods: We detected the luminescence signal by using lab-made optical imaging equipment of animal light imaging system (ALIS) and different two kinds of light sources. One is three bacterial light-emitting sources containing different number of bacteria. The other is three different non-bacterial light sources emitting very weak light. By using the concept of the candela and the flux, we could derive simplified linear quantification formula. After experimentally measuring light intensity, the data was processed with the proposed quantification function. Results: We could obtain linear response of photon counts to measurement time by applying the pre-determined quantification function. The ratio of the re-calculated photon counts and measurement time present a constant value although different light source was applied. Conclusion: The quantification function for linear response could be applicable to the standard quantification process. The proposed method could be used for the exact quantitative analysis in various light imaging equipments with presenting linear response behavior of constant light emitting sources to measurement time.