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Study on the Safety of Firefly Luciferase in Human as a Transient Reporter Gene of Oncolytic Virotherapy (항암 바이러스 치료제의 보고유전자로써 반딧불이 루시퍼레이즈의 인체 내 안전성에 대한 연구)

  • Hong, Young Mi;Yoon, Woong Hee;Lee, You Ra;Kim, Soo Ji;Ngabire, Daniel;Narayanasamy, Badrinath;Ornella, Mefotse Saha Cyrelle;Kim, Myunghee;Cho, Euna;Lee, Bora;Hwang, Tae-Ho
    • Journal of Life Science
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    • v.31 no.11
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    • pp.1028-1036
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    • 2021
  • Firefly luciferase (FLuc) can function as an efficient marker in the gene and viral therapies. Nonetheless, its clinical translation has been unaccomplished with the concerns on its exogenous nature and the similarity with human fatty acyl-CoA synthetase. In this study, we aimed to show safety of FLuc by conducting a set of preclinical experiments and a human use. Initially, FLuc permeability across the plasma membrane was investigated by delivering the FLuc-carrying viral vector, OTS-412, or the FLuc recombinant protein. After in vitro infection of OTS-412 into different cancer cell lines, FLuc activity was detected only in the cell lysates, but not in culture media. In addition, recombinant FLuc protein further showed the impermeability against the plasma membrane. Similar result was also observed in the in vivo experiment. After being injected into the VX2 tumor-bearing rabbit, the FLuc exclusively resided within the tumor tissue without being detected in the blood plasma or other organs. Human cancer cell lines originated from various organs were lysed and treated to the FLuc, and none of the human substrates was reactive against the FLuc. As a final step, FLuc recombinant protein was intravenously injected into a human. The luciferase was degraded with the half-life of 20 to 30 minutes in blood, and was untraceable from 1.5 hr after the injection. In addition, the blood plasma was nonresponsive against the fatty acids. Hematological analysis was also comparable between the pre- and post-injection. Altogether, our study collectively demonstrates the safety of the firefly luciferase.

A Study on the Dynamics of Dissolved Organic Matter Associated with Ambient Biophysicochemical Factors in the Sediment Control Dam (Lake Youngju) (영주댐 유사조절지 상류의 용존유기물 (Dissolved Organic Matter) 특성과 물리·화학 및 생물학적 환경 요인과의 연관성 연구)

  • Oh, Hye-Ji;Kim, Dokyun;Choi, Jisoo;Chae, Yeon-Ji;Oh, Jong Min;Shin, Kyung-Hoon;Choi, Kwangsoon;Kim, Dong-Kyun;Chang, Kwang-Hyeon
    • Korean Journal of Ecology and Environment
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    • v.54 no.4
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    • pp.346-362
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    • 2021
  • A sediment control dam is an artificial structure built to prolong sedimentation in the main dam by reducing the inflow of suspended solids. These dams can affect changes in dissolved organic matter (DOM) in the water body by changing the river flow regime. The main DOM component for Yeongju Dam sediment control of the Naeseongcheon River was analyzed through 3D excitation-emission matrix (EEM) and parallel factor (PARAFAC) analyses. As a result, four humic-like components (C1~C3, C5), and three proteins, tryptophan-like components (C2, C6~C7) were detected. Among DOM components, humic-like components (autochthonous: C1, allochthonous: C2~C3) were found to be dominant during the sampling period. The total amount of DOM components and the composition ratio of each component did not show a difference for each depth according to the amount of available light (100%, 12%, and 1%). Throughout the study period, the allochthonous organic matter was continuously decomposing and converting into autochthonous organic matter; the DOM indices (fluorescence index, humification index, and freshness index) indicated the dominance of autochthonous organic matter in the river. Considering the relative abundance of cyanobacteria and that the number of bacteria cells and rotifers increased as autochthonous organic matter increased, it was suggested that the algal bloom and consequent activation of the microbial food web was affected by the composition of DOM in the water body. Research on DOM characteristics is important not only for water quality management but also for understanding the cycling of matter through microbial food web activity.

Biological Toxicity Assessment of Sediment at an Ocean Dumping Site in Korea (폐기물 배출해역 퇴적물의 생물학적 독성평가 연구)

  • Seok, Hyeong Ju;Kim, Young Ryun;Kim, Tae Won;Hwang, Choul-Hee;Son, Min Ho;Choi, Ki-young;Kim, Chang-joon
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.28 no.1
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    • pp.1-9
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    • 2022
  • The effect of sediments in a waste dumping area on marine organisms was evaluated using sediment toxicity tests with a benthic amphipod (Monocorophium acherusicum) and bioluminescent bacterium (Vibrio fischeri) in accordance with the Korean Standard Method for Marine Wastes (KSMMW). Nine sites in the East Sea-Byeong, East Sea-Jeong, and Yellow Sea-Byeong areas were sampled from 2016 to 2019. The test results showed that the relative average survival rate (benthic amphipods) and relative luminescence inhibition rate (luminescent bacteria) were below 30%, which were judged to be "non-toxic." However, in the t-test, a total of 12 benthic amphipod samples (6, 1, 1, and 4 in 2016, 2017, 2018, and 2019, respectively) were significantly different (p<0.05) from the control samples. To identify the source of toxicity on benthic amphipods, a simple linear regression analysis was performed between the levels of eight heavy metals (Cr, As, Ni, Cd, Cu, Pb, Zn, and Hg) in sediments and the relative average survival rate. The results indicated that Cr had the highest contribution to the toxicity of benthic amphipods (p = 0.000, R2 = 0.355). In addition, Cr was detected at the highest concentration at the DB-85 station and exceeded the Marine Environment Standards every year. Although the sediments were determined as "not toxic" according to the ecotoxicity criteria of the KSMMW, the results of the statistical significance tests and toxicity identification evaluation indicated that the toxic effect was not acceptable. Therefore, revising the criteria for determining the toxic effect by deriving a reference value through quantitative risk assessment using species sensitivity distribution curves is necessary in the future.

Apoptotic Effect of Proso Millet Grains on Human Breast Cancer MDA-MB-231Cells Is Exerted by Activation of BAK and BAX, and Mitochondrial Damage-mediated Caspase Cascade Activation (기장 종자 유래 추출물의 인간 유방암 MDA-MB-231 세포에 대한 세포독성에 관련된 미토콘드리아 손상-의존적 아폽토시스 유도 효과)

  • Do Youn Jun;Cho Rong Han;Young Ho Kim
    • Journal of Life Science
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    • v.33 no.1
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    • pp.15-24
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    • 2023
  • To examine the antitumor effect of proso millet grains, whether proso millet grains exert apoptotic activity against human cancer cells was investigated. When the cytotoxicity of 80% ethanol (EtOH) extract of proso millet grains was tested against various cancer cells using MTT assay, more potent cytotoxicity was observed against human breast cancer MDA-MB-231 cells than against other cancer cells. When the EtOH extract was evaporated to dryness, dissolved in water, and then further fractionated by sequential extraction using four organic solvents (n-hexane, methylene chloride, ethyl acetate, and n-butanol), the BuOH fraction exhibited the highest cytotoxicity against MDA-MB-231 cells. Along with the cytotoxicity, TUNEL-positive apoptotic nucleosomal DNA fragmentation and several apoptotic responses including BAK/BAX activation, mitochondria membrane potential (Δψm) loss, mitochondrial cytochrome c release into the cytosol, activation of caspase-8/-9/-3, and degradation of poly (ADP-ribose) polymerase (PARP) were detected. However, human normal mammary epithelial MCF-10A cells exhibited a significantly lesser extent of sensitivity compared to malignant MDA-MB-231 cells. Irrespective of Fas-associated death domain (FADD)-deficiency or caspase-8-deficiency, human T acute lymphoblastic leukemia Jurkat cells displayed similar sensitivities to the cytotoxicity of BuOH fraction, excluding an involvement of extrinsic apoptotic mechanism in the apoptosis induction. These results demonstrate that the cytotoxicity of BuOH fraction from proso millet grains against human breast cancer MDA-MB-231 cells is attributable to intrinsic apoptotic cell death resulting from BAK/BAX activation, and subsequent mediation of mitochondrial damage-dependent activation of caspase cascade.

Development of cordycepin fortified meat production in Hanwoo steers I. Determination of the chemical composition and safety of Cordyceps militaris from grain or pupae and Paecilomyces tenuipes (Cordycepin 강화 한우고기 생산에 관한 연구 I. 곡립기주 동충하초 균사체의 화학적 성분 및 안전성 조사)

  • Kim, W.Y.;Lee, S.H.;Kim, D.H.;Lee, J.H.;Nho, W.G.;Hwang, J.H.;Yeo, J.M.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.11 no.1
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    • pp.35-51
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    • 2009
  • Three different Cordyceps militaris(Paecilomyces tenuipes and Cordyceps militaris from grain or pupae) were analysed for their chemical composition, fatty acids composition and the concentration of amino acids, cordycepin and heavy metal to obtain basic information on them as feed supplements for livestock. The crude protein content of C. militaris from grain and pupae was 76.16 and 75.45%, respectively, being higher than that of P. tenuipes(57.21%). The concentration of linoleic acid was much higher but that of linolenic acid was much lower for C. militaris from grain than for the others. The cordycepin content was significantly higher for C. militaris from grain than for that from pupae (1.64% vs 0.68% on a DM basis). But cordycepin was not detected in P. tenuipes. Heavy metal contents(Cr, Pb, Cd, Hg) for all C. militaris were below the allowance levels recommended by Ministry of Agriculture and Forestry in Republic of Korea.

Evaluation of Physical Properties and Material Characterization for Structural Frame at the Stained Glass Windows to Gongju Jeil Church of the Registered Cultural Heritage in Korea (국가등록문화재 공주제일교회 스테인드글라스 구조재의 재질특성과 물성 평가)

  • Bo Young Park;Hye Ri Yang;Chan Hee Lee
    • Economic and Environmental Geology
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    • v.56 no.1
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    • pp.103-114
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    • 2023
  • The Christian Museum of Gongju Jeil Church was first built in 1931 and was largely damaged during the Korean War, but the walls and chimneys have been preserved. This building has a high architectural values in that the chapel was reconstructed in 1956, and maintains its original form through repair of damaged parts rather than new construction. The stained glass windows were as installed in 1979 and has a great significance in the Dalle de Verre method using lump glass. However, some of the stained glass damaged partially, such as various cracks and splits, and vertical and horizontal cracks in the joint fillers of supporting the colored glass. As the structural materials of the stained glass window, an iron frame and cement mortar filled with it were used, and corrosion of iron, cracking of mortar and granular decomposition appear partially due to weathering. In the joint fillers, the content of Ca and S is very high, indicating that gypsum were used as admixtures, and the gypsums grow in a rhombohedral and forms a bundle, which is investigated to have undergone recrystallization. As a result of modeling the ultrasonic velocity at the joint fillers, the left and right windows at the entrance show relatively weak in the range of 800 to 1,600m/s, and the lower right corner of the altar window and the upper left corner of the center window were also 1,000 to 1,800m/s, showing relatively low physical properties. And gypsums produced during the neutralization of lime mortar were detected in the joint fillers and contaminants on the surface. Such salts may cause damage to the joint material due to freezing and thawing, so appropriate preventive conservation is required. Also, since various damage types are complexly appearing in stained glass window and joint filler, customized conservation treatment should be reviewed through clinical tests.

Antioxidant Activity of Tea Made from Korean Mountain-Cultivated Ginseng Leaves and its Influence on Lipid Metabolism (장뇌삼 엽차의 항산화활성 및 지질대사에 미치는 영향)

  • Bae, Man-Jong;Kim, Soo-Jung;Ye, Eun-Ju;Nam, Hak-Sik;Park, Eun-Mi
    • Journal of the Korean Society of Food Culture
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    • v.24 no.1
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    • pp.77-83
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    • 2009
  • This study was conducted to investigate the chemical composition and biological function of tea made from mountain-cultivated ginseng leaves. The antioxidant activities of tea made from mountain-cultivated ginseng leaves were determined by measuring their electron-donating ability based on their DPPH and nitrite-scavenging ability. The electron-donating abilities of tea made from mountain-cultivated ginseng leaves (500 and 1,000 ppm) as determined by DPPH assay were 45.6 and 85.1%, respectively. The nitrite scavenging ability of tea made from mountain-cultivated ginseng leaves (500 and 1,000 ppm) at pH 6.0 were 32.8 and 51.4%, respectively. Furthermore, the nitrite scavenging activity increased in a dose-dependent manner at all pH values. The effects of tea made from mountain-cultivated ginseng leaves on Male Sprague-Dawley rats were also evaluated. To accomplish this, the rats were divided into three groups (A: normal diet group, B: high fat diet group and C: high fat diet supplemented with tea made from mountain-cultivated ginseng leaves group). The anti-obesity effects of tea made from mountain-cultivated ginseng leaves were then evaluated. The serum total lipid, total cholesterol and triglyceride contents in C group were lower than those of B group; however, these differences were not statistically significant. The HDL-cholesterol content was significantly higher in the C group than in the other groups. Taken together the results of this study suggest that tea made from mountain-cultivated ginseng leaves possesses antioxidant activity and improves lipid metabolism.

A study of analytical method for Benzo[a]pyrene in edible oils (식용유지 중 벤조피렌 분석법 비교 연구)

  • Min-Jeong Kim;jun-Young Park;Min-Ju Kim;Eun-Young Jo;Mi-Young Park;Nan-Sook Han;Sook-Nam Hwang
    • Analytical Science and Technology
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    • v.36 no.6
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    • pp.291-299
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    • 2023
  • The benzo[a]pyrene in edible oils is extracted using methods such as Liquid-liquid, soxhlet and ultrasound-assisted extraction. However these extraction methods have significant drawbacks, such as long extraction time and large amount of solvent usage. To overcome these drawbacks, this study attempted to improve the current complex benzo[a]pyrene analysis method by applying the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method that can be analyzed in a simple and short time. The QuEChERS method applied in this study includes extraction of benzo[a]pyrene into n-hexane saturated acetonitrile and n-hexane. After extraction and distribution using magnesium sulfate and sodium chloride, benzo[a]pyrene is analyzed by liquid chromatography with fluorescence detector (LC/FLR). As a result of method validation of the new method, the limit of detection (LOD) and quantification (LOQ) were 0.02 ㎍/kg and 0.05 ㎍/kg, respectively. The calibration curves were constructed using five levels (0.1~10 ㎍/kg) and coefficient (R2) was above 0.99. Mean recovery ratio was ranged from 74.5 to 79.3 % with a relative standard deviation (RSD) between 0.52 to 1.58 %. The accuracy and precision were 72.6~79.4 % and 0.14~7.20 %, respectively. All results satisfied the criteria ranges requested in the Food Safety Evaluation Department guidelines (2016) and AOAC official method of analysis (2023). Therefore, the analysis method presented in this study was a relatively simple pretreatment method compared to the existing analysis method, which reduced the analysis time and solvent use to 92 % and 96 %, respectively.

Diagnostic Performance of Digital Breast Tomosynthesis with the Two-Dimensional Synthesized Mammogram for Suspicious Breast Microcalcifications Compared to Full-Field Digital Mammography in Stereotactic Breast Biopsy (정위적 유방 조직검사 시 미세석회화 의심 병변에서의 디지털 유방단층영상합성법과 전역 디지털 유방촬영술의 진단능 비교)

  • Jiwon Shin;Ok Hee Woo;Hye Seon Shin;Sung Eun Song;Kyu Ran Cho;Bo Kyoung Seo
    • Journal of the Korean Society of Radiology
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    • v.83 no.5
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    • pp.1090-1103
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    • 2022
  • Purpose To evaluate the diagnostic performance of digital breast tomosynthesis (DBT) with the two-dimensional synthesized mammogram (2DSM), compared to full-field digital mammography (FFDM), for suspicious microcalcifications in the breast ahead of stereotactic biopsy and to assess the diagnostic image visibility of the images. Materials and Methods This retrospective study involved 189 patients with microcalcifications, which were histopathologically verified by stereotactic breast biopsy, who underwent DBT with 2DSM and FFDM between January 8, 2015, and January 20, 2020. Two radiologists assessed all cases of microcalcifications based on Breast Imaging Reporting and Data System (BI-RADS) independently. They were blinded to the histopathologic outcome and additionally evaluated lesion visibility using a fivepoint scoring scale. Results Overall, the inter-observer agreement was excellent (0.9559). Under the setting of category 4A as negative due to the low possibility of malignancy and to avoid the dilution of malignancy criteria in our study, McNemar tests confirmed no significant difference between the performances of the two modalities in detecting microcalcifications with a high potential for malignancy (4B, 4C, or 5; p = 0.1573); however, the tests showed a significant difference between their performances in detecting microcalcifications with a high potential for benignancy (4A; p = 0.0009). DBT with 2DSM demonstrated superior visibility and diagnostic performance than FFDM in dense breasts. Conclusion DBT with 2DSM is superior to FFDM in terms of total diagnostic accuracy and lesion visibility for benign microcalcifications in dense breasts. This study suggests a promising role for DBT with 2DSM as an accommodating tool for stereotactic biopsy in female with dense breasts and suspicious breast microcalcifications.

Improving the Nutritional Value of Tenebrio molitor Larvae by Feeding Them a Soymilk Residue-added Food Source (두유박 첨가 먹이원 급이에 따른 갈색거저리 유충의 영양성 증진 효과)

  • So-Yun Kim;Min Ji Park;Jeong-Hun Song;Sangmin Ji;Gyu-Dong Chang;Sun Young Kim
    • Journal of Life Science
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    • v.34 no.3
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    • pp.191-198
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    • 2024
  • To verify the nutritional value and safety of larvae fed with a soymilk residue-added food source, we compared and analyzed the nutritional components of and harmful substances in Tenebrio molitor larvae reared on wheat bran. Crude protein content based on dry weight was 1.2 times higher in the 10% soymilk residue-fed group (SR) (54.0%) than in the wheat bran-fed group (WB) (43.5%). Dietary fiber also tended to be 1.9 times more in the SR (4.9%) than the WB (2.5%). Among unsaturated fatty acids, the linoleic acid content was found to be 1.1-fold higher in the SR (32.9%) than in the WB (29.0%). Potassium, which was the most abundant among the macro minerals, was 1.1-fold more abundant in the WB (1,074.5 mg/100 g) than in the SR (1,014.0 mg/100 g). Among the micro minerals, zinc content was 1.2-fold higher in the SR (14.5 mg/100 g) than in the WB (11.9 mg/100 g). The results of the analysis of hazardous substances in the WB and SR revealed that the amount of heavy metals met the standards for heavy metals in edible insects, and food poisoning-inducing bacteria such as Escherichia coli and Salmonella spp. were not detected in all groups. These results indicate that mealworm larvae fed with 10% soymilk residue have abundant nutrients and are safe for intake. Thus, food sources with added soymilk residue has the potential to be used as feed ingredients.