성장배지 혈액 유무가 구강미생물의 적색 형광 발현에 미치는 영향

정승화 ( Seung-hwa Jeong ),양용훈 ( Yong-hoon Yang ),이민아 ( Min-ah Lee ),김세연 ( Se-yeon Kim ),김지수 ( Ji-soo Kim ) 대한예방치과·구강보건학회 2017 大韓口腔保健學會誌 Vol.41 No.4

Objectives: Dental plaque emits red fluorescence under a visible blue light near the ultra-violet end of the light spectrum. The fluorescence characteristics of each microorganism have been reported in several studies. The aim of this study was to evaluate changes in red fluorescence of oral microorganisms that is affected by blood in the culture media. Methods: The gram-positive Actinomyces naeslundii (AN, KCTC 5525) and Lactobacillus casei (LC, KCTC 3109) and gram negative Prevotella intermedia (PI, KCTC 3692) that are known to emit red fluorescence were used in this study. Each bacterium was activated in broth and cultivated in different agar media at 37℃ for 7 days. Tryptic soy agar with hemin and vitamin K₃ (TSA), TSA with sheep blood (TSAB), basal medium mucin (BMM) medium, and BMM with sheep blood (BMMB) were used in this study. Fluorescence due to bacterial growth was observed under 405-nm wavelength blue light using the quantitative light-induced fluorescence-digital (QLF-D) device. The red, green, and blue fluorescence values of colonies were obtained using image-analysis software and the red to green ratio (R/G value) and red to total RGB ratio (R/RGB value) were calculated for quantitative comparison. Results: The QLF-D images of the AN, LC, and PI colonies showed red fluorescence in all media, but the fluorescence of all bacteria was reduced in TSA and BMM media, compared with in TSAB and BMMB media. Both the R/G and the R/RGB values of all bacteria were significantly reduced in growth media without blood (P<0.001). Conclusions: Based on this in vitro study, it can be concluded that red fluorescence of oral bacteria can be affected by growth components, especially blood. Blood-containing medium could be a significant factor influencing red fluorescence of oral bacteria. It can be further hypothesized that bleeding in the oral cavity can increase the red fluorescence of dental plaque.

레이저 형광법을 이용한 인접면 우식증의 진단

설재헌,오유향,이난영,이상호 大韓小兒齒科學會 2004 大韓小兒齒科學會誌 Vol.31 No.2

레이저 형광법을 이용하여 초기 인접면 우식증을 탐지할 수 있는지의 여부와 그 탐지 감도를 평가하여 인접면 우식증의 진단에 활용 가능한지의 여부를 규명하기 위하여 사람의 치아를 사용하여 다양한 깊이의 인공우식병소를 유발시키고 이를 가시광선 투과법에 의한 사진, 교익 방사선사진 촬영, 레이저 형광법, 광활성 염료를 이용한 레이저 형광법 등으로 관찰하여 검사법 간의 일치도, 상관관계, 병소 깊이에 따른 광밀도 등을 분석하여 다음과 같은 결과를 얻었다. 1. 탈회시간과 각 검사법들과 일치도 검사에서 사진의 경우 tau-c 값이 0.08로 탈회시간에 따른 사진의 검사 수치가 일치하지 않았으나 교익 방사선사진, 레이저 형광법, 광활성 염료를 이용한 레이저 형광법의 tau-c 값은 각각 0.60, 0.48, 0.64로 일치하는 것으로 나타났다. 2. 탈회시간에 따른 병소깊이와 각 검사법 간의 상관관계는 광활성 염료를 이용한 레이저 형광법(r=0.51), 레이저 형광법(r=0.43), 교익 방사선사진(r=0.35), 사진(r=0.33) 순으로 높았으며 광활성 염료를 이용한 레이저 형광법과 레이저 형광법은 탈회 시간과 상관관계가 있었다(P<0.05). 3. 교익 방사선사진을 기준 검사법으로 한 레이저 형광법과 광활성 염료를 이용한 레이저 형광법의 진단학적 민감도는 각각 67%, 100%였으며 특이도는 57%, 11%로 민감도에 비해 상대적으로 낮게 나타났다. 4. 병소의 깊이에 따른 병소 표면에서의 건전 치질과 우식 치질 사이의 광밀도의 차이(DFR)는 광활성 염료를 이용한 레이저 형광법이 레이저 형광법에 비해 크게 나타났으며(P<O.05) 병소 깊이의 변화에 따른 광밀도 변화는 레이저 형광법의 경우 유의한 변화를 보이는데 반해 광활성 염료를 이용한 레이저 형광법에서는 변화를 보이지 않았다. 이상의 결과를 종합하면, 초기 인접면 우식증의 진단에 있어 레이저 형광법과 광활성 염료를 이용한 레이저 형광법을 교익 방사선사진에 뒤지지 않은 진단능을 가지고 있으나 우식 병소의 정성적인 분석뿐 아니라 정량적 분석이 가능한지의 여부는 향후 더 많은 연구가 필요하리라 사료된다. Artificial carious lesions in various depths were observed with visual examination using light transillumination, bite-wing radiography, laser fluorescence, and dye-enhanced laser fluorescence to determine the reproducibility, correlation of each diagnostic method, diagnostic sensitivity and diagnostic specificity. And optical densities according to demineralized times were measured whether laser fluorescence could be used as a quantitative diagnostic method. The following results were obtained whether laser fluorescence could be used for diagnosis of initial proximal caries. 1. Tau-c values of visual examination was 0.08 which showed lowest reproducibility, and those of bite-wing radiography, laser fluorescence, dye-enhanced laser fluorescence were 0.60, 0.48, and 0.64. respectively which showed relatively high reproducibility. 2. The correlation between demineralization time and each examination was the highest in dye-enhanced laser fluorescence(γ=0.51), followed by laser fluorescence(γ=0.43), bite-wing radiograph(γ=0.35), and visual examination (γ= 0.33). Dye-enhanced laser fluorescence and laser fluorescence showed significant correlation with demineralization time. 3. The sensitivity of laser fluorescence and dye-enhanced laser fluorescence for diagnosing approximal caries based on bite-wing radiography were 67%, 100% and those of specificity were 57%. 11% which showed diagnostic specificity was relatively lower than sensitivity. 4. The difference in optical density(DFR) between sound teeth and carious lesions according to lesion depth was high with dye-enhanced laser fluorescence compared with laser fluorescence. DFR measured with laser fluorescence according to changes in lesion depth was statistically significant but was not statistically significant with dye-enhanced laser fluorescence. Based on these results, laser fluorescence and dye-enhanced laser fluorescence have comparable diagnostic power as bite-wing radiography in early diagnosis of proximal caries.

Experience Profiling of Fluorescence-Guided Surgery I: Gliomas

( So Young Ji ),( Jin Wook Kim ),( Chul-kee Park ) 대한뇌종양학회·대한신경종양학회·대한소아뇌종양학회 2019 Brain Tumor Research and Treatment Vol.7 No.2

Background Numerous studies reported a usefulness of 5-aminolevulinic acid (5-ALA) fluorescence- guided surgery (FGS) in high grade gliomas. However, fluorescence patterns and intensities are variable among gliomas. In this study, we report our extensive experience with FGS in various gliomas, focusing on epidemiological data of fluorescence patterns. Methods A total of 827 histologically proven glioma patients out of 900 brain tumor patients who had undergone FGS using 5-ALA during the period of 8.5 years between July 2010 and January 2019 were analyzed. Indications of FGS in glioma surgery are evidence for possible high-grade foci in putative gliomas in preoperative MRI. Results Among the 827 gliomas, the number of cases corresponding to 2016 World Health Organization (WHO) grade IV, III, II, and I are 528 (58.7%), 193 (21.4%), 87 (9.7%) and 19 (2.1%), respectively. In terms of fluorescence rate, grade IV gliomas showed positive fluorescence in 95.4% of cases including strong intensity in 85.6%. Grade III gliomas showed fluorescence in about half of cases (55.0%), but 45.0% of the cases showed no fluorescence at all. Anaplastic oligodendroglioma had a higher positive rate (63.9%) than anaplastic astrocytoma (46.2%). Both grade II and I gliomas still showed positive fluorescence in about one-fourth of cases (24.1% and 26.3% respectively). Among them ependymoma and pilocytic astrocytoma were fluorescence-prone tumors. Conclusion This epidemiological data of 5-ALA fluorescence in various grades of glioma provides a basic reference to the clinical application of FGS with 5-ALA in glioma surgery.

Fluorescence change of Fusobacterium nucleatum due to Porphyromonas gingivalis

이민아,강시묵,김세연,김지수,김진범,정승화 한국미생물학회 2018 The journal of microbiology Vol.56 No.9

The aim of this study was to measure changes in the fluorescence of Fusobacterium nucleatum interacting with Porphyromonas gingivalis for excitation with blue light at 405-nm. P. gingivalis was mono- and co-cultivated in close proximity with F. nucleatum. The fluorescence of the bacterial colonies was photographed using a QLF-D (Quantitative Light-induced Fluorescence-Digital) Biluminator camera system with a 405 nm light source and a specific filter. The red, green and blue intensities of fluorescence images were analyzed using the image analysis software. A fluorescence spectrometer was used to detect porphyrin synthesized by each bacterium. F. nucleatum, which emitted green fluorescence in single cultures, showed intense red fluorescence when it was grown in close proximity with P. gingivalis. F. nucleatum co-cultivated with P. gingivalis showed the same pattern of fluorescence peaks as for protoporphyrin IX in the red part of the spectrum. We conclude that the green fluorescence of F. nucleatum can change to red fluorescence in the presence of adjacent co-cultured with P. gingivalis, indicating that the fluorescence character of each bacterium might depend on the presence of other bacteria.

Fluorescence Quenching of Green Fluorescent Protein during Denaturation by Guanidine

Hackjin Kim*,Jaebok Park,맹필재,Kichul Jung 대한화학회 2005 Bulletin of the Korean Chemical Society Vol.26 No.3

Fluorescence of green fluorescent protein mutant, 2-5 GFP is observed during denaturation by guanidine. The fluorescence intensity decreases exponentially but the fluorescence lifetime does not change during denaturation. The fluorescence lifetime of the denatured protein is shorter than that of native form. As the protein structure is modified by guanidine, solvent water molecules penetrate into the protein barrel and protonate the chromophore to quench fluorescence. Most fluorescence quenchers do not affect the fluorescence of native form but accelerate the fluorescence intensity decay during denaturation. Based on the observations, a simple model is suggested for the structural change of the protein molecule during denaturation.

Development of a fluorescence-image scoring system for assessing noncavitated occlusal caries

Jung, Eun-Ha,Lee, Eun-Song,Jung, Hoi-In,Kang, Si-Mook,de Josselin de Jong, Elbert,Kim, Baek-Il Elsevier 2018 Photodiagnosis and photodynamic therapy Vol.21 No.-

<P><B>Abstract</B></P> <P><B>Background</B></P> <P>This study aimed (1) to develop a scoring system based on a quantitative light-induced fluorescence (QLF) score for the occlusal caries (QS-Occlusal) that standardizes the fluorescence properties of noncavitated lesions from QLF images, (2) to confirm the validity and reliability of QS-Occlusal, and (3) to determine whether it is possible to replace existing clinical examinations by image evaluations based on the developed QS-Occlusal for assessing occlusal caries lesions.</P> <P><B>Methods</B></P> <P>This clinical study investigated 791 teeth of 94 subjects. The teeth were assessed by visual and tactile examinations using ICDAS criteria and quantitative light-induced fluorescence-digital (QLF-D) image examinations. QS-Occlusal was divided into four stages (from 0 to 3) based on the progression level of the lesion and the fluorescence loss and red fluorescence on captured QLF-D images. Two trained examiners who were not involved in the visual examination evaluated occlusal fluorescence images using QS-Occlusal. The maximum loss of fluorescence (|Δ<I>F</I> <SUB>max</SUB>|) and the maximum change in the ratio of red and green fluorescence (Δ<I>R</I> <SUB>max</SUB>) were quantitatively analyzed by the QA2 software to detect differences between the QS-Occlusal groups. The modalities were compared in terms of sensitivity, specificity, and area under the receiver operating characteristics (AUROC) curve for three different thresholds of the ICDAS codes: 0 vs 1–4 (D<SUB>1</SUB>), 0–2 vs 3/4 (D<SUB>2</SUB>), and 0–3 vs 4 (D<SUB>3</SUB>).</P> <P><B>Results</B></P> <P>|Δ<I>F</I> <SUB>max</SUB>| increased significantly by about 4.7-fold (from 15.94 to 75.63) when QS-Occlusal increased from 0 to 3. Δ<I>R</I> <SUB>max</SUB> was about 6.2-fold higher for QS-Occlusal=1 (49.74) than for QS-Occlusal=0 (8.04), and 21.6-fold higher for QS-Occlusal=3 (<I>P<</I> 0.05). The new QS-Occlusal showed an excellent AUROC (ranging from 0.807 to 0.976) in detecting occlusal caries when optimum cutoff values were applied. The intra- and interexaminer agreements indicated excellent reliability, with ICC values of 0.94 and 0.86, respectively.</P> <P><B>Conclusions</B></P> <P>The QS-Occlusal proposed in this study can be used in the clinical detection of noncavitated lesions with an excellent diagnostic ability. This makes it possible to replace clinical examinations and intuitively evaluate the lesion severity and status relatively easily and objectively by applying this scoring system to fluorescence images.</P> <P><B>Highlights</B></P> <P> <UL> <LI> QLF technology can be a useful screening tool for noncavitated occlusal lesion. </LI> <LI> QLF score can be used to evaluate the severity and status of occlusal caries lesion nondestructively. </LI> <LI> QLF score may be able to replace existing clincal examination in the future with excellent diagnostic ability for detecting noncavitated caries. </LI> </UL> </P>

Experience Profiling of Fluorescence-Guided Surgery II: Non-Glioma Pathologies

( So Young Ji ),( Jin Wook Kim ),( Chul-kee Park ) 대한뇌종양학회·대한신경종양학회·대한소아뇌종양학회 2019 Brain Tumor Research and Treatment Vol.7 No.2

Background Only sporadic reports of fluorescence-guided surgery (FGS) have been published for non-glioma conditions. In this study, we focus on epidemiological data of fluorescence patterns and report the diverse experiences of FGS in non-gliomas. Methods During 8.5 years between July 2010 and January 2019, 900 FGS for brain tumor performed in Seoul National University Hospital. Among them, a total of 73 histologically proven nonglioma patients were analyzed. Indications for FGS have been the possibility of anaplastic tumor in intra- axial tumors in preoperative MRI and an attempt to reproduce known anecdotal experiences of 5-Aminolevulinic Acid (5-ALA) fluorescence. Results In cases of brain tumors except for gliomas, the most frequent cases were brain metastasis (23 cases) followed by lymphomas (9 cases) and meningeal tumors (8 cases). And there were embryonal tumors (6 cases), hemangioblastomas (4 cases), and solitary fibrous tumor/hemangiopericytomas (3 cases). Most brain metastases, meningiomas, primary central nervous system lymphomas, and treatment effect cases showed positive fluorescence. Moreover, some non-tumorous conditions also showed positive fluorescence. However, hemangioblastoma and germ cell tumor did not observe any fluorescence at all. Conclusion 5-ALA induced fluorescence is not limited to glioma but is also evident in non-glioma and non-neoplastic conditions. This 5-ALA-induced fluorescence may be used as an intraoperative tool for various brain conditions.

설태의 형광특성 - 설태 형광현상의 발현기전 소개 및 제안 -

김지혜 ( Ji-hye Kim ),남동현 ( Dong-hyun Nam ) 대한한의진단학회 2011 大韓韓醫診斷學會誌 Vol.15 No.1

In traditional Korean medicine, inspection of the tongue is an important method of making medical diagnoses and determining prognosis. We surveyed the fluorescence characteristics of the tongue coat in the ultraviolet light. The tongue coat comprises micro-organisms, blood metabolites, leukocytes from periodontal pockets, large amounts of desquamated epithelial cells released from the oral mucosa and different nutrients. In the ultraviolet light tissues of the oral cavity generally emit weak red or green fluorescence, which is not easily seen by the human eye, but is readily detected. This fluorescence has been proved to be due to the production of porphyrins by oral micro-organisms. While the composition of motile micro-organisms on the dorsum of the tongue is not constant, variations also occur persistingly in the fluorescence characteristics of the tongue coat. But because live bacteria contain a variety of intracellular biomolecules that have specific excitation and emission wavelength spectra characterizing their intrinsic fluorescence, the tongue coat emits fluorescence. the tongue itself, on the other hand, emits very weak or not fluorescence. In conclusion, we suggests that the uncoated tongue area be eliminated from the coated tongue area with the difference between the fluorescence characteristics of the tongue and that of the tongue coat.

Fluorescence Quenching of Green Fluorescent Protein during Denaturation by Guanidine

Jung, Ki-Chul,Park, Jae-Bok,Maeng, Pil-Jae,Kim, Hack-Jin Korean Chemical Society 2005 Bulletin of the Korean Chemical Society Vol.26 No.3

Fluorescence of green fluorescent protein mutant, 2-5 GFP is observed during denaturation by guanidine. The fluorescence intensity decreases exponentially but the fluorescence lifetime does not change during denaturation. The fluorescence lifetime of the denatured protein is shorter than that of native form. As the protein structure is modified by guanidine, solvent water molecules penetrate into the protein barrel and protonate the chromophore to quench fluorescence. Most fluorescence quenchers do not affect the fluorescence of native form but accelerate the fluorescence intensity decay during denaturation. Based on the observations, a simple model is suggested for the structural change of the protein molecule during denaturation.

Use of Chlorophyll a Fluorescence Imaging for Photochemical Stress Assessment in Maize (Zea mays L.) Leaf under Hot Air Condition

Jong Yong Park,Sung Young Yoo,Hong Gyu Kang,Tae Wan Kim 한국작물학회 2016 한국작물학회지 Vol.61 No.4

The objective of this study was to find a rapid determination of the hot air stress in maize (Zea mays L.) leaves using a portable chlorophyll fluorescence imaging instrument. To assess the photosynthetic activity of maize leaves, an imaging analysis of the photochemical responses of maize was performed with chlorophyll fluorescence camera. The observed chlorophyll imaging photos were numerically transformed to the photochemical parameters on the basis of chlorophyll a fluorescence. Chlorophyll a fluorescence imaging (CFI) method showed that a rapid decrease in maximum fluorescence intensity (Fm) of leaf occurred under hot air stress. Although no change was observed in the maximum quantum yield (Fv/Fm) of the hot air stressed maize leaves, the other photochemical parameters such as maximum fluorescence intensity (Fm) and Maximum fluorescence value (Fp) were relatively lowered after hot air stress. In hot air stressed maize leaves, an increase was observed in the nonphotoquenching (NPQ) and decrease in the effective quantum yield of photochemical energy conversion in photosystem II (Φ PSII). Thus, NPQ and ΦPSII were available to be determined non-destructively in maize leaves under hot air stress. Our results clearly indicated that the hot air could be a source of stress in maize leaves. Thus, the CFI analysis along with its related parameters can be used as a rapid indicating technique for the determining hot air stress in plants.