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      • 세포주에 따른 담배연기응축물의 소핵생성 비교

        신한재,손형옥,이영구,이동욱,현학철 한국연초학회 2003 한국연초학회지 Vol.25 No.2

        Although tobacco smoke has been known to have genotoxicity as well as cytotoxicity, the sensitivity of the cell lines used against cigarette smoke is poorly understood. The objective of this study was to evaluate and compare the genotoxicity of several cell lines, which are routinely used in the in vitro assays, with cigarette smoke condensate(CSC) of Kentucky Reference Cigarette 1R4F. In the micronucleus(MN) induction assays, murine(CHO-K1, V79, BALB/c 3T3) cell lines and human(MCF-7, A549) ones were used. As a result, the CSC exhibited cytotoxicity with a concentration-dependent response in all cell lines. EC$_{50}$ of CSC in CHO-K1, V79, BALB/c 3T3, MCF-7 and A549 were 140, 125, 100, 116 and 109 $\mu\textrm{g}$/mL, respectively. On the other hand, the spontaneous micronucleated cell(MNC) frequency was stable and reproducible in every cell lines tested in this study. The dose-response of various cell lines to the induction of MN by CSC was estimated using linear regression analysis. CSC(0~100 $\mu\textrm{g}$/mL) caused a dose-dependent MN induction in CHO-K1, V79, BALB/c 3T3 and MCF-7 cell lines. Putting together all the data obtained and linear regression analysis of the data, we concluded that V79 cells are more susceptible to the accurate assessment of CSC-induced MN than the others.s.

      • KCI등재

        Comparative In Vitro Biological Toxicity of Four Kinds of Air Pollution Particles

        신한재,조현기,박창균,박기홍,임흥빈 한국독성학회 2017 Toxicological Research Vol.33 No.4

        Accumulating epidemiological evidence indicates that exposure to fine air pollution particles (APPs) is associated with a variety of adverse health effects. However, the exact physiochemical properties and biological toxicities of fine APPs are still not well characterized. We collected four types of fine particle (FP) (diesel exhaust particles [DEPs], natural organic combustion [NOC] ash, synthetic organic combustion [SOC] ash, and yellow sand dust [YSD]) and investigated their physicochemical properties and in vitro biological toxicity. DEPs were almost entirely composed of ultrafine particles (UFPs), while the NOC, SOC, and YSD particles were a mixture of UFPs and FPs. The main elements in the DEPs, NOC ash, SOC ash, and YSD were black carbon, silicon, black carbon, and silicon, respectively. DEPs exhibited dose-dependent mutagenicity even at a low dose in Salmonella typhimurium TA 98 and 100 strains in an Ames test for genotoxicity. However, NOC, SOC, and YSD particles did not show any mutagenicity at high doses. The neutral red uptake assay to test cell viability revealed that DEPs showed dose-dependent potent cytotoxicity even at a low concentration. The toxicity of DEPs was relatively higher than that of NOC, SOC, and YSD particles. Therefore, these results indicate that among the four FPs, DEPs showed the highest in vitro biological toxicity. Additional comprehensive research studies such as chemical analysis and in vivo acute and chronic inhalation toxicity tests are necessary to determine and clarify the effects of this air contaminant on human health.

      • 담배적용 생체외(in vitro) 평가

        신한재,박철훈,Shin, Han-Jae,Park, Chul-Hoon 한국연초학회 2008 한국연초학회지 Vol.30 No.1

        세포독성과 유전독성 측정법은 독성 평가에 있어서 통합요소이며, 잠재적인 생물학적 활성을 측정하는 방법으로서 규제 당국에 의해 인정되는 방법이다. 이들은 빠르고 경제적인 방법이고, 오랜 사용한 역사를 가지고 있으며, 양적인 독성을 평가 할 수 있는 방법이다. 담배의 안전성을 평가하기위해 널리 이용되는 3-battery 시험법은 다음과 같다. 1. 미생물을 이용한 복귀돌연변이 시험 2. 포유류 유래 세포주를 이용한 Neutral red 세포독성 3. 포유류 유래 세포주를 이용한 소핵시험

      • 담배 주류연의 생물학적 활성에 대한 흡연조건의 영향

        신한재,박철훈,손형옥,이형석,유지혜,이병찬,현학철,Shin, Han-Jae,Park, Chul-Hoon,Sohn, Hyung-Ok,Lee, Hyeong-Seok,Yoo, Ji-Hye,Lee, Byeong-Chan,Hyun, Hak-Chul 한국연초학회 2008 한국연초학회지 Vol.30 No.1

        The objective of this study was to determine the effect of smoking conditions on the in vitro toxicological activity of mainstream smoke. The 2R4F reference cigarette was machine-smoked by International Organization for Standardization(ISO) and Canadian Intense(CI) conditions. Smoke was analysed for chemical composition and in vitro toxicity. The cytotoxic potencies of both the total particulate matter(TPM), which were collected in Cambridge filter pad, and gas/vapor phase(GVP), which was bubbled through in phosphate-buffer saline in a gas-washing bottle, were assessed neutral red up take assay with chinese hamster ovary(CHO) cells. The assessment for genotoxicity of TPMs generated under ISO and CI conditions was determined using Salmonella mutagenicity assay and in vitro micronucleus assay. When calculated on an equal TPM basis, in vitro toxicity of TPM obtained under CI condition was decreased compared to TPM generated under ISO condition. The results of chemical composition analyses revealed that the lower toxicological activity under CI condition than that of ISO condition could be explained by the decreased in the contents of phenols, N-nitrosoamines and aromatic amines of TPM on an equal TPM basis.

      • 35종의 특정 화학성분들의 in vitro 활성 평가

        신한재,손형옥,박철훈,이형석,민영근,현학철,Shin, Han-Jae,Sohn, Hyung-Ok,Park, Chul-Hoon,Lee, Hyeong-Seok,Min, Young-Keun,Hyun, Hak-Chul 한국연초학회 2007 한국연초학회지 Vol.29 No.1

        The objective of this study was to investigate the contribution of various smoke constituents to the toxicological activity of total particulate matter(TPM) or the gas/vapor phase(GVP). These components included phenol compounds, aromatic amines, polycyclic aromatic hydrocarbons, heterocyclic amines, and carbonyl compounds. The mutagenic and cytotoxic potencies were assessed using the Salmonella mutagenicity assay with S. typimurium TA98 strain and the neutral red uptake cytotoxicity assay(NRU) with BALB/c 3T3 fibroblast cells, respectively. The Salmonella mutagenicity test showed that heterocyclic amines exhibited significantly higher levels of toxicity compared to other smoke constituents. Among them, 2-amino-3,4-dimethylimidazo[4,5-f]quinoline(MeIQ) was shown the most mutagenic compound with a specific mutagenicity of $7.9{\times}10^5\;revertants/{\mu}g$. An analysis of the possible contribution revealed that MeIQ account for only 0.85% of the 2R4F-TPM mutagenicity in TA98. NRU data demonstrated that high cytotoxic activity was obtained for hydroquinone, formaldehyde, and acrolein. Based on the results of the present study, the contribution of acrolein to the cytotoxicity of the GVP fraction was calculated as 61%. Thus, a large proportion of the cytotoxic activity of this complex mixture, cigarette smoke gas phase, can be attributed to the acrolein.

      • 질소 화합물이 담배연기성분 및 안전성에 미치는 영향

        신한재,박철훈,손형옥,이형석,김용하,현학철,Shin, Han-Jae,Park, Chul-Hoon,Sohn, Hyung-Ok,Lee, Hyeong-Seok,Kim, Yong-Ha,Hyun, Hak-Chul 한국연초학회 2009 한국연초학회지 Vol.31 No.2

        The objective of this study was to investigate the effect of nitrogen compounds such as protein on the chemical composition and toxicity of cigarette mainstream smoke. BSA protein was treated into the tobacco leaf of original 2R4F cigarette at 1~4 % level. The studies were performed which included a bacterial mutagenicity assay and a mammalian cell cytotoxicity assay for total particulate matter(TPM), and glutathione(GSH) consumption assay for gas/vapor phase(GVP) and determination of smoke chemical constitute. Cigarettes treated with protein were observed dose-dependent increase in yield of volatiles, semi-volatiles and aromatic amines compared with control cigarette. However, carbonyl compounds such as acrolein was lower than that of control cigarette when calculated on an equal TPM basis. The cytotoxicity of TPM obtained from the protein-added cigarettes was not different from that of control cigarette. However, the mutagenicity of the TPM from protein-treated cigarettes(1~4 %) was up to 10-27 % higher than that of control. On the other hand, toxicity of GVP from protein-treated cigarette(4 %) was significantly decreased compared with control cigarette. An overall assessment of our data suggests that nitrogen compounds such as protein should be important for the chemical composition and biological activity of cigarette mainstream smoke.

      • 담배연기 중 생물학적 활성 성분

        신한재,Shin, Han-Jae 한국연초학회 2010 한국연초학회지 Vol.32 No.1

        담배 주류연은 수천가지 종류의 다양한 형태의 화학물질로 구성되어있고, 이들 화학물질의 대부분은 극히 미량의 농도로 존재하는 것으로 알려져 있다(Green and Rodgman, 1996). 담배연기 속에서 발암물질로서는 poly cyclic aromatic hydrycarbon (PAHs) 류인 B(a)P를 최초로 보고된 이후, 현재까지 69종의 화합물이 담배연기중의 발암물질로서 알려졌으며, 여기에는 9종의 PAHs 그리고 4종의 aromatic amine, nitrosoamines 및 aldehydes 그리고 다른 무기 및 유기 화합물들을 포함된다. 또한 담배 주류연에는 여러 화학물질들에 의해 유발된 생물학적 독성을 상당히 감소시키는 것으로 확인된 성분들이 존재하는 것으로 알려져 있다. 이러한 담배연기 중에 존재하는 항 발암성 효과를 나타내는 성분들의 작용 메카니즘은 발암성 물질 또는 발암성 물질의 대사산물들과 반응을 하거나, 불활성화 시키거나, 세포 구성 물질들과의 반응에서 서로 경쟁하거나, 또는 발암성의 흡수를 간섭함으로서 발암성 효과를 없애거나 또는 감소시키는 것으로 생각된다. 여러 연구자들은 담배연기 응축물에서 종양 성장의 억제 작용을 하는 성분들이 존재하는 것으로 주장 하고 있다.(Hoffmann and Griffin, 1958; Fall et al., 1964; Homburger and Tregier, 1965). 담배 연기 중에는 발암성을 나타내는 성분들과 함께 항 발암성 및 항 돌연변이성 효과를 나타내는 성분들이 존재한다는 것이 다양한 생물학적 평가 연구들에 의해 증명되었다(Slaga and DiGivanni, 1984; Fay et al., 1984; Green and Rodgman, 1996).

      • 담배주류연의 세포독성에 대한 담배필터의 영향

        신한재,손현옥,한정호,박철훈,허재연,이동욱,황건중,현학철,Shin Han-Jae,Sohn Hyung-Ok,Han Jung-Ho,Park Chul-Hoon,Hur Jae-Yeon,Lee Dong-Wook,Hwang Keon-Joong,Hyun Hak-Chul 한국연초학회 2005 한국연초학회지 Vol.27 No.1

        The objective of this study was to evaluate the effect of cigarette filter on in vitro cytotoxicity of cigarette mainstream smoke from the cigarette. In this work, we used 3 types of cigarettes included non-filtered 2R4F cigarette, cellulose acetate-filtered 2R4F cigarette, and carbon dual-filtered 2R4F cigarette which was made from original 2R4F by replacing with an acetate filter containing carbon. The cytotoxicity of both the cigarette smoke condensate (CSC), which was collected in Cambridge filter pad, and the gas/vapor phase (GVP), which was bubbled through in phosphate-buffered saline in a gas-washing bottle, was determined using a neutral red uptake assay with CHO-K1 cells. With regard to cytotoxicity when calculated on an equal puff basis, the cytotoxicity of CSC from the filtered cigarettes was lower than that of the non filtered cigarette. Also, $EC_{50}$ vlaue of GVP from carbon filter cigarette was 40.9 puff/L, indicating the cytotoxicity to be $20\%$ lower than that of the CA filter cigarette. The cytotoxicity of the GVP was correlated to the several vapor phase components (formaldehyde, acetaldehyde, acetone, acrolein, crotonaldehyde and MEK). In conclusion, carbon filter, which significantly reduced the amount of carbonyl compounds in mainstream cigarette smoke, results in significant reductions in the cytotoxicity potential of the smoke.

      • KCI등재

        3종의 배암차즈기에 대한 염증억제 활성 비교 및 활성성분 구명

        신한재,곽효민,장미,박시환,민혜정,이정민,이문용,김종한,김성원,한창균,김승형 한국약용작물학회 2016 韓國藥用作物學會誌 Vol.24 No.5

        Background: Salvia has been widely cultivated for use in flavoring and folk medicines in many countries, including Korea and China. In this study, we investigated the anti-inflammatory activity and the underlying active compounds of Salvia extract and its fractions.Methods and Results: The anti-inflammatory activity was measured by assessing the inhibition of cysteinyl leukotriene production in rat basophilic leukemia (RBL)-2H3 mast cells. Salvia plebeia R. Br. was found to have the most potent inhibitory activity on leukotriene production than S. japonica and S. chanroenica had. Fifty percent ethanol extracts of S. plebeia R. Br. were successively partitioned with n-hexane, methylene chloride, ethyl acetate, 1-butanol and water. The ethyl acetate (EtOAc) fraction showed stronger anti-inflammatory activity than other solvent fractions did. The EtOAc fraction was subjected to silica gel column chromatography elution with a chloroform and methanol gradient system (100 : 1 → 1 : 1) yielding 10 fractions. Three kinds of fractions (chloroform:methanol = 20 : 1, 10 : 1 and 5 : 1) showed high inhibitory activity on leukotriene production. We confirmed the major compounds with anti-inflammatory activity from S. plebeia R. Br.Conclusions: In this study, the major components of S. plebeia that showed leukotriene production inhibitory activity were isolated using solvent extraction and silica gel column chromatography. Rosmarinic acid, hispidulin and luteolin were identified as the major compounds with anti-inflammatory effect.

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