• Nuclear Medicine and Molecular Imaging
• /
• v.43 no.5
• /
• pp.451-458
• /
• 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.

• Journal of Food Hygiene and Safety
• /
• v.34 no.3
• /
• pp.290-295
• /
• 2019

Salmonella spp. are frequently associated with food and are among the most important foodborne pathogens. The recent Salmonella out breaks in Korea was associated with chocolate mousse cakes served with school meals during September 2018. The objective of this research was to compare the 3M Molecular Detection Assay 2 - Salmonella and the Korean Standard Method of Salmonella in artificially inoculated mousse (chocolate and cheese) and tiramisu cakes. Mousse (chocolate and cheese) and tiramisu cakes were artificially inoculated with S. Typhimurium. Twenty five gram of sample was enriched with 225 mL buffered peptone water for incubation at $37^{\circ}C$ for 24 h. After enrichment, the cultures were analyzed by using the 3M Molecular Detection Assay 2 - Salmonella and the Korean Standard Method. Most of the inoculated samples showed similar results except the chocolate mousse cakes, in which real-time PCR was unable to detect S. Typhimurium even after $10^4CFU/25g$ of inoculation. However, S. Typhimurium inoculated at a concentration of $10^0CFU/25g$ was detected by using 3M Molecular Detection Assay 2 - Salmonella. In chocolate mousse, detection of S. Typhimurium using real-time PCR was partially successful when dark chocolate was added at less than 15%. Negative results in real-time PCR and 3M Molecular Detection Assay 2 - Salmonella were confirmed by gel electrophoresis. The data indicated that dark chocolate could inhibit amplification of the target gene in the PCR reactions. In conclusion, the 3M Molecular Detection Assay 2 - Salmonella was better than the Korean Standard Method (real-time PCR) for the detection of S. Typhimurium in chocolate mousse cakes and chocolate mousse.

• Journal of Digital Convergence
• /
• v.19 no.1
• /
• pp.211-216
• /
• 2021

This study was carried out to evaluate the effects of disinfection using environmental disinfectant after having assessed the extent of contamination through microbial culture testing and the Adenosine Triphosphate Bioluminescence method among the environmental management evaluation methods used for the environment in the hospital ward of patients infected by multidrug-resistant organisms. This study was conducted with the patient wards isolated due to multidrug-resistant organisms as the environmental surface. Specimens were collected from five locations including infusion pumps, IV poles, bedside cabinets, bed railings, keyboards, and blood pressure measurement cuffs. ATP and microbial culture testing were executed prior to, immediately after, and five minutes post-disinfection. According to the result contamination of the infusion pumps was statistically significantly reduced after disinfection. In addition, the bacteria before and after disinfection reduced in IV pole, bed railing, and keyboard. That is, regular environmental surface disinfection can provide safer environments to patients against infection. Therefore, it is necessary to establish guidelines including disinfection methods and intervals for environmental surfaces by evaluating the persistence of disinfectants at various institutions in the future.

• ALGAE
• /
• v.37 no.2
• /
• pp.149-161
• /
• 2022

Noctiluca scintillans is a heterotrophic dinoflagellate that causes red-colored oceans during the day (red tides) and glowing oceans at night (bioluminescence). This species feeds on diverse prey, including phytoplankton, heterotrophic protists, and eggs of metazoans. Thus, many scientists have conducted studies on the ecophysiology of this species. It is easy to cultivate N. scintillans at a scale of <1 L, but it is difficult to cultivate them at a scale of >100 L because N. scintillans cells usually stay near the surface, while prey cells stay below the surface in large water tanks. To obtain mass-cultured N. scintillans cells, we developed an automatic system for cultivating N. scintillans on a scale of 100 L. The system consisted of four tanks containing fresh nutrients, the chlorophyte Dunaliella salina as prey, N. scintillans for growth, and N. scintillans for storage, respectively. The light intensities supporting the high growth rates of D. salina and N. scintillans were 300 and 20 µmol photons m^-2 s^-1, respectively. Twenty liters of D. salina culture from the prey culture tank were transferred to the predator culture tank, and subsequently 20 L of nutrients from the nutrient tank were transferred to the prey culture tank every 2 d. When the volume of N. scintillans in the predator culture tank reached 90 L 6 d later, 70 L of the culture were transferred to the predator storage tank. To prevent N. scintillans cells from being separated from D. salina cells in the predator culture tank, the culture was mixed using an air pump, a sparger, and a stirrer. The highest abundance of N. scintillans in the predator culture tank was 45 cells mL^-1, which was more than twice the highest abundance when this dinoflagellate was cultivated manually. This automatic system supplies 100 L of N. scintillans pure culture with a high density every 10 d for diverse experiments on N. scintillans.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.5 no.6
• /
• pp.395-399
• /
• 2000

Five different freeze-dried recombinant bioluminescent bacteria were used for the detection of cellular stresses caused by endocrine disrupting chemicals. These strains were DPD2794 (recA::luxCDABE), which is sensitive to DNA damage, DPD2540 (fabA::luxCDABE), sensitive to cellular membrane damage, DPD2511 (katG::luxCDABE), sensitive to oxidative damage, and TV1061 (grpE::luxCDABE), sensitive to protein damage. GC2, which emits bioluminescence constitutively, was also used in this study. The toxicity of several chemicals was measured using GC2. Damage caused by known endocrine disrupting chemicals, such as nonyl phenol, bisphenol A, and styrene, was detected and classified according to toxicity mode, while others, such as phathalate and DDT, were not detected with the bacteria. These results suggest that endocrine disrupting chemicals are toxic in bacteria, and do not act via an estrogenic effect, and that toxicity monitoring and classification of some endocrine disrupting chemicals may be possible in the field using these freeze-dried recombinant bioluminescent bacteria.

• Bulletin of the Korean Chemical Society
• /
• v.34 no.6
• /
• pp.1673-1678
• /
• 2013

Lumazine protein is a member of the riboflavin synthase superfamily and the intense fluorescence is caused by non-covalently bound to 6,7-dimethyl 8-ribityllumazine. To figure out the binding modes and the structure of the N-terminal domain of lumazine protein, the wild type of protein extending to amino acid 118 (N-LumP 118 Wt) and mutants of N-LumP 118 V41W, S48W, T50W, D64W, and A66W from Photobacterium leiognathi were purified. The biochemical properties of the wild type and mutants of N-LumP 118 proteins were analyzed by absorbance and fluorescence spectroscope. The peak of absorbance and fluorescence of lumazine ligand were shifted to longer wavelength on binding to N-LumPs. The observed absorbance value at 410 nm of lumazine bound to N-LumP 118 proteins indicate that one mole of N-LumP 118 proteins bind to one mole of ligand of lumazine. Fluorescence analysis show that the maximum peak of fluorescence of N-LumP S48W was shifted to the longest wavelength by binding with 6,7-dimethyl 8-ribityllumazine and was shown to the greatest quench effect by acrylamide among all tryptophan mutants.

• Food Science of Animal Resources
• /
• v.38 no.4
• /
• pp.816-822
• /
• 2018

Bifidobacterium is recognized as one of the most beneficial microorganisms in our gut. Many researches on bifidobacteria have been done to understand their roles in the gut. The objective of the present study was to develop a bioluminescent labelling plasmid vector for bifidobacteria to facilitate their visualization in vitro, in situ, and in vivo. A plasmid replicon (2.0 kb) of plasmid pFI2576 previously identified from B. longum FI10564 was amplified by PCR and cloned into pUC19 plasmid vector (2.68 kb). The cloned replicon was subcloned into pTG262 ($luc^+$) recombinant plasmid vector (7.4 kb) where a luciferase gene ($luc^+$) from pLuc2 (8.5 kb), an Escherichia coli and lactobacilli shuttle vector, was inserted into pTG262 plasmid vector. The final recombinant DNA, pTG262::pFI2576 rep ($luc^+$), was transferred into a B. catenulatum strain. This recombinant strain showed 3,024 relative luminescence units at $OD_{600}$ value of 0.352. Thus, this recombinant plasmid construct can be broadly used for labelling bifidobacteria.

• 한국생물공학회:학술대회논문집
• /
• 2000.04a
• /
• pp.117-120
• /
• 2000

For detecting toxicity of endocrine disruptors (EDs), rapid, sensitive, and simple methods are needed. Therefore, in this study, a new method in which the different toxic effect of EDs can be monitored using 4 different recombinant bacteria was designed and evaluated. It was found that the recombinant bacteria could monitor the toxic effect, not estrogenic effect, due to EDCs through the measurement of bioluminescence and cell growth rate, which were shown to depend upon a form of cellular toxicity, such as DNA damage, protein damage, oxidative damage, and membrane damage. In addition, it was found that the damage done by EDCs can be divided into several groups based upon the toxic mechanisms of the EDCs

• Proceedings of the Korean Society of Sericultural Science Conference
• /
• 2003.10a
• /
• pp.136-137
• /
• 2003

Fireflies, the luminescent insect, have species specific flash patterns, being recognized as sexual communication. The lucifrrase gene is sole enzyme responsible for bioluminescence. The firefly luciferase gene is widely used as a genetic marker or as a reporter gene in a variety of organism including bacteria, plants and animals. In this study, we illustrate the complete organization of the genomic structure of the luciferase gene from L. lateralis sampled in Boun and Muju, Korea. (omitted)

• Journal of Microbiology
• /
• v.38 no.3
• /
• pp.150-155
• /
• 2000

A bioluminescent assay method for detecting the activity of phospholipase C(PLC; phosphatidyl choline cholinephosphohydrolase, EC 3.1.4.3) was developed using bioluminescent marine bacteria. Phospholipase C from Bacillus cereus and sn-1,2- dimyristoyl glycerol was further hydrolyzed with lipase from Candida ecylidracea. The hydrolyzed myristic acid was quantified using a dark mutant of Vibrio harveyi (designated as M-17). The in vivo light intensity of which was stimulated specifically up to one thousand fold in the presence of myristic acid. The rates of the hdrolysis of the DMPC substrate by the phospholipase measured by the luminescence method were linear with time and the were estalished to detect as little as 0.1 mUnit of phospholipase C and 5 nM of myristic acid production.