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Study on hydroxy fatty acid contents changes and physiological responses under abiotic stresses in transgenic Camelina

  • Kim, Hyun-Sung;Lee, Hyun-Sook;Lim, Hyun-Gyu;Park, Won;Kim, Hyun-Uk;Lee, Kyeong-Ryeol;Ahn, Sung-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.191-191
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    • 2017
  • Hydroxy fatty acid (HFA) is an important industrial resource that known to be extracted from seeds of Castor or Lesquerella. However, mass production of HFA from those crops are difficult because of their behavior or life cycle. In this study, we applied HFA synthesis related gene FAH12, RcPDAT1, RcLPCAT, RcDGAT2, and RcPDCT on bioenergy crop Camelina sativa. Furthermore, we determined NaCl or cold stress tolerance changes of transgenic Camelina. RcFAH12, RcPDAT1, RcLPCAT, RcDGAT2, and RcPDCT genes were cloned into multigene expression vector which is engineered with seed specific promoter of FAE1 or Napin. Combination of HFA genes multi-expression vector constructs were divided into Set3 (RcFAH12, RcPDAT1-2, RcLPCAT), Set4 (RcFAH12, RcDGAT2, RCPDAT1-2, RcLPCAT), and Set5 (RcFAH12, RcDGAT2, RCPDAT1-2, RcLPCAT, RcPDCT). Transgenic HFA synthesis Camelina plants were generated using agrobacterium-mediated vacuum infiltration system. Results of fatty acid composition of T1 transgenic Camelina seeds analyzed by GC-MS showed 9.5, 9.0, and 13.6 % of HFA contents in Set3#6, Set4#8, and Set5#10, respectively. Therefore, seeds of T2 generation were harvest from Set5#10 which is shown highest HFA contents, and, 17.7, 8.1 and 10.5 % of HFA contents were determined in Set5#10-5, Set5#10-8, and Set#10-10, respectively. However, 7.7% of C18:2 and 22.3 % of C18:3 among unsaturated fatty acids were decreased in Set5#10-5 than WT. Meanwhile, we confirmed abiotic stress responses in T2 transgenic Camelina Set5#10-5 and Set5#10-10 under 0, 100, 150, and 200 mM NaCl or 25, 15, and $10^{\circ}C$ temperature for 5 weeks. Both Set5#10-5 and Set5#10-10 showed lower growth in height than WT in control and NaCl condition. Growth of leaf length and width were similar in WT and Set5#10-10 but lower in Set5#10-5 under NaCl stress. Number of opened flowers showed that both transgenic Camelina were lower than WT under normal condition. But, WT and Set5#10-10 showed similar opened flower number in 100 and 200 mM NaCl. In cold stress, 15 and $10^{\circ}C$ treatment for 5 weeks did not showed significant changes in between WT and both transgenic lines even they showed different growth rate in control condition. Taken together, growth and development are delayed by expression of exogenous HFA related genes in transgenic lines but relative abiotic stress sensitivity is similar with WT. In conclusion, reduced C18:2 or C18:3 fatty acid composition of seed by HFA synthesis is resulted from lack of resource supplement for development at seedling stage but it is not affect NaCl and cold stress tolerance.

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Development of an Effective Method for Testing Resistance to Black Spot of Radish Caused by Alternaria brassicicola (Alternaria brassicicola에 의한 무 검은무늬병에 대한 효율적인 저항성 검정법 개발)

  • Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Hun;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.35 no.2
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    • pp.210-219
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    • 2017
  • This study was conducted to establish an efficient screening method for radish (Raphanus sativus) cultivars that are resistant to black spot, which is caused by Alternaria brassicicola. Seven A. brassicicola isolates were selected and investigated for their ability to produce spores and pathogenicity. Of these isolates, A. brassicicola KACC 40036 and 43923 produced abundant spores in V-8 juice agar medium and showed pathogenicity and strong virulence on radish seedlings. We examined the resistance of 61 commercial cultivars of radish to A. brassicicola KACC40036, and found that there are no highly resistant radish cultivars; however, some cultivars, such as 'Geumbong' and 'Searom', showed weak resistance to A. brassicicola. For further study, we selected four radish cultivars that showed different disease responses to A. brassicicola KACC40036. According to the growth stage of the radish seedlings, inoculum concentration, and incubation temperature of radish, development of black spot on four cultivars has been investigated. The results showed that younger seedlings were more sensitive to A. brassicicola than older seedlings, and the disease severity depended on the concentration of the spore suspension. The disease severity of plants incubated in humidity chamber at $25^{\circ}C$ was greater than that of plants grown at $20^{\circ}C$ or $30^{\circ}C$. Taken together, we suggest the following method for screening for radish plants that are resistant to A. brassicicola: 1) inoculate 16-day-old radish seedlings with an A. brassicicola spore suspension ($2.0{\times}10^5spores{\cdot}mL^{-1}$) using the spray method, 2) incubate the inoculated plants in a humidity chamber at $25^{\circ}C$ for 24 h and then transfer the plants to a growth chamber at $25^{\circ}C$ with 80% relative humidity under a 12 h light/dark cycle, and 3) assess the disease severity of the plants two days after inoculation.

The Effect of Epigallocatechin-3-gallate on HIF-1 α and VEGF in Human Lung Cancer Cell Line (비소세포폐암주에서 저산소상태에 의해 유발된 HIFa-1 α와 VEGF의 발현증가에 미치는 Epigallocatechin-3-gallate의 억제 효과)

  • Song, Joo Han;Jeon, Eun Joo;Kwak, Hee Won;Lee, Hye Min;Cho, Sung Gun;Kang, Hyung Koo;Park, Sung Woon;Lee, Jae Hee;Lee, Byung Ook;Jung, Jae Woo;Choi, Jae Cheol;Shin, Jong Wook;Kim, Ki Jeong;Kim, Jae-Yeol;Park, In Won;Choi, Byoung Whui
    • Tuberculosis and Respiratory Diseases
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    • v.66 no.3
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    • pp.178-185
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    • 2009
  • Background: Epigallocatechin-3-gallate (EGCG) is the major catechin in green tea, and has shown antiproliferative, antiangiogenic, antimetastatic and cell cycle pertubation activity in various tumor models. Hypoxia can be induced because angiogenesis is insufficient for highly proliferating cancer. Hypoxia-inducible factor-1$\alpha$ (HIF-1$\alpha$) and its downstream target, vascular endothelial growth factor (VEGF), are important for angiogenesis, tumor growth and metastasis. The aim of this study was to determine how hypoxia could cause changes in the cellular phenomena and microenvironment in a non-small cell culture system and to examine the effects of EGCG on a HIF-1$\alpha$ and VEGF in A549 cell line. Methods: A549 cells, a non-small cell lung cancer cell line, were cultured with DMEM and 10% fetal bovine serum. A decrease in oxygen tension was induced using a hypoxia microchamber and a $CO_2-N_2$ gas mixture. Gas analysis and a MTT assay were performed. The A549 cells were treated with EGCG (0, 12.5, 25, 50 ${\mu}mol/L$), and then examined by real-time-PCR analysis of HIF-1$\alpha$, VEGF, and $\beta$-actin mRNA. Results: Hypoxia reduced the proliferation of A549 cells from normoxic conditions. EGCG inhibited HIF-1$\alpha$ transcription in A549 cells in a dose-dependent manner. Compared to HIF-1$\alpha$, VEGF was not inhibited by EGCG. Conclusion: HIF-1$\alpha$ can be inhibited by EGCG. This suggests that targeting HIF-1$\alpha$ with a EGCG treatment may have therapeutic potential in non-small cell lung cancers.

Microbial Community Composition Associated with Anaerobic Oxidation of Methane in Gas Hydrate-Bearing Sediments in the Ulleung Basin, East Sea (동해 울릉분지 가스 하이드레이트 매장 지역의 메탄산화 미생물 군집 조성 및 분포)

  • Cho, Hyeyoun;Kim, Sung-Han;Shin, Kyung-Hoon;Bahk, Jang-Jun;Hyun, Jung-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.20 no.1
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    • pp.53-62
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    • 2015
  • To elucidate the microbial consortia responsible for the anaerobic methane oxidation in the methane hydrate bearing sediments, we compared the geochemical constituents of the sediment, the rate of sulfate reduction, and microbial biomass and diversity using an analysis of functional genes associated with the anaerobic methane oxidation and sulfate reduction between chimney site (UBGH2-3) on the continental slope and non-chimney site (UBGH2-10) on the basin of the Ulleung Basin. From the vertical profiles of geochemical constituents, sulfate and methane transition zone (SMTZ) was clearly defined between 0.5 and 1.5 mbsf (meters below seafloor) in the UBGH2-3, and between 6 and 7 mbsf at the UBGH2-10. At the UBGH2-3, the sulfate reduction rate (SRR) in the SMTZ exhibited was appeared to be $1.82nmol\;cm^{-3}d^{-1}$ at the depth of 1.15 mbsf. The SRR in the UBHG2-10 showed a highest value ($4.29nmol\;cm^{-3}d^{-1}$) at the SMTZ. The 16S rRNA gene copy numbers of total Prokaryotes, mcrA, (methyl coenzyme M reductase subunit A), and dsrA (dissimilatory sulfite reductase subunit A) showed the peaks in the SMTZ at both sites, but the maximum mcrA gene copy number of the UBGH2-10 appeared below the SMTZ (9.8 mbsf). ANME-1 was a predominant ANME (Anaerobic MEthanotroph) group in both SMTZs of the UBGH2-3 and -10. However, The sequences of ANME-2 were detected only at 2.2 mbsf of the UBGH2-3 where high methane flux was observed because of massive amount of gas hydrate at shallow depth. And Desulfosarcina-Desulfococcus (DSS) that is associated with ANME-2 was detected in 2.2 mbsf of the UBHG2-3. Overall results demonstrate that ANME-1 and ANME-2 are considered as significant archaeal groups related to methane cycle in the subsurface sediment of the East Sea, and ANME-2/DSS consortia might be more responsible for methane oxidation in the methane seeping region than in non-seeping region.

Evaluation of the Temperature Drop Effect and the Rutting Resistance of Moisture Retaining-Porous Asphalt Pavement Using Accelerated Pavement Testing (포장가속시험을 이용한 보수형 배수성 포장의 온도저감 효과 및 소성변형 저항특성 연구)

  • Kwak, Byoung-Seok;Suh, Young-Chan;Song, Chul-Young;Kim, Ju-Won
    • International Journal of Highway Engineering
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    • v.11 no.3
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    • pp.97-109
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    • 2009
  • One of the main causes of asphalt rutting is high temperature of the pavement. Nevertheless, there has been few research on lowering the pavement temperature for reducing rutting. This study investigated the performance characteristics of moisture-retaining porous asphalt pavement, which is known to have a temperature reducing effect. The purpose of this study is to quantify the temperature reducing effect of moisture-retaining porous asphalt pavement and its effect of reducing rutting through Accelerated Pavement Testing(APT). Additionally, the possibility of reducing the thickness of the pavement in comparison to general dense grade pavement by analyzing structural layer coefficient of moisture retaining pavement. A total of three test sections consisting of two moisture-retaining porous asphalt pavement sections and one general dense-grade porous asphalt pavement section were constructed for this study. Heating and spraying of water were carried out in a regular cycle. The loading condition was 8.2 ton of wheel load, the tire pressure of $7.03kgf/cm^2$, and the contact area of $610cm^2$. The result of this experiment revealed that the temperature reducing effect of the pavement was about $6.6{\sim}7.9^{\circ}C$(average of $7.4^{\circ}C$) for the middle layer and $7.9{\sim}9.8^{\circ}C$(average of $8.8^{\circ}C$) for surface course, resulting in a rutting reduction of 26% at the pavement surface. Additionally, the structural layer coefficient of moisture retaining pavement measured from a laboratory test was 0.173, about 1.2 times that of general dense grade pavement. The general dense-grade porous asphalt pavement test section exhibited rutting at all layers of surface course, middle layer, and base layer, while the test sections of moisture-retaining porous asphalt pavement manifested rutting mostly at surface course only.

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Adsorption Removal of Sr by Barium Impregnated 4A Zeolite (BaA) From High Radioactive Seawater Waste (Barium이 함침된 4A 제올라이트 (BaA)에 의한 고방사성해수폐액에서 Sr의 흡착 제거)

  • Lee, Eil-Hee;Lee, Keun-Young;Kim, Kwang-Wook;Kim, Ik-Soo;Chung, Dong-Yong;Moon, Jei-Kwon;Choi, Jong-Won
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.14 no.2
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    • pp.101-112
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    • 2016
  • This study investigated the removal of Sr, which was one of the high radioactive nuclides, by adsorption with Barium (Ba) impregnated 4A zeolite (BaA) from high-radioactive seawater waste (HSW). Adsorption of Sr by BaA (BaA-Sr), in the impregnated Ba concentration of above 20.2wt%, was decreased by increasing the impregnated Ba concentration, and the impregnated Ba concentration was suitable at 20.2wt%. The BaA-Sr adsorption was added to the co-precipitation of Sr with $BaSO_4$ precipitation in the adsorption of Sr by 4A (4A-Sr) within BaA. Thus, it was possible to remove Sr more than 99% at m/V (adsorbent weight/solution volume)=5 g/L for BaA and m/V >20 g/L for 4A, respectively, in the Sr concentration of less than 0.2 mg/L (actual concentration level of Sr in HSW). It shows that BaA-Sr adsorption is better than 4A-Sr adsorption in for the removal capacity of Sr per unit gram of adsorbent, and the reduction of the secondary solid waste generation (spent adsorbent etc.). Also, BaA-Sr adsorption was more excellent removal capacity of Sr in the seawater waste than distilled water. Therefore, it seems to be effective for the direct removal of Sr from HSW. On the other hand, the adsorption of Cs by BaA (BaA-Cs) was mainly performed by 4A within BaA. Accordingly, it seems to be little effect of impregnated Ba into BaA. Meanwhile, BaA-Sr adsorption kinetics could be expressed the pseudo-second order rate equation. By increasing the initial Sr concentrations and the ratios of V/m, the adsorption rate constants ($k_2$) were decreased, but the equilibrium adsorption capacities ($q_e$) were increasing. However, with increasing the temperature of solution, $k_2$ was conversely increased, and $q_e$ was decreased. The activation energy of BaA-Sr adsorption was 38 kJ/mol. Thus, the chemical adsorption seems to be dominant rather than physical adsorption, although it is not a chemisorption with strong bonding form.

Effect of Scrapping Aerial Mycelia and Light on the Production of Macroconidia and Chlamydospores of Cylindrocarpon destructans Causing Root Rot of Panax ginseng (기중균사 제거와 광처리가 인삼 뿌리썩음병균 Cylindrocarpon destructans의 대형분생포자 및 후막포자 생성에 미치는 영향)

  • Cho Dae-Hui;Yu Yun-Hyun;Ohh Seung-Hwan
    • Journal of Ginseng Research
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    • v.23 no.3 s.55
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    • pp.123-129
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    • 1999
  • Under the light condition of 25,000 Lux (12 hrs dark and light cycle) with scrapping treatment of aerial mycelia of Cylindrocarpon destructans on potato dextrose agar (PDA), V-8 juice agar, and ginseng extract agar, production of the macroconidia was increased to $3.7\~8.1$ fold over them produced in the dark. They were also produced $7.7\~18.0$ times more in the liquid cultures under the light condition than under the dark as well. PDA and V-8 juice agar among the tested were the best for the macroconidium production. On PDA, 1,585 $macroconidia/mm^2$ were produced under the light of 25,000 Lux with scrapping treatment of aerial mycelia of C. destructans, which is 3.2 and 1.4 times more than those produced under 3,000 and 10,000 Lux, respectively. Meanwhile, $20\~99$ macroconidia/$mm^2$ were produced by the non-scrapping under the light condition between 3,000 Lux and 25,000 Lux. The macroconidia were, however, lysed at $6\~7$ days after being incubated under the above range of the light. They were consisted of $1\~3$ cells in a macroconidium while $69.4\~100\%$ of them were the two-celled and the number did not seem to be affected by either the scrapping or the light. Production of chlamydospore converted from mycelia of C. destructans seemed to be promoted by the light and the scrapping as well. The 1,285 chlamydospres/$mm^2$ were produced with the light (25,000 Lux), which is 2.8 and 1.2 times more than those with 3,000 and 10,000 Lux, respectively. Scrapping the aerial mycelia of the cultures increased the chlamydospore formation to 1.9, 2.5 and 1.4 times more than the non-scrapping under the light intensity of 3,000 Lux, 10,000 Lux, and 25,000 Lux, respectively. On PDA, 1 to 8 chlamydospore(s) per catena were formed by all treatments tested and $34.2\~58.9\%$ of them was a single chlamydospore, However, the numbers was affected by neither the light ($3,000\~25,000$ Lux) nor the scrapping the aerial mycelia.

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The Generation of Westerly Waves by Sobaek Mountains (소백산맥에 의한 서풍 파동 발생)

  • Kim, Jin wook;Youn, Daeok
    • Journal of the Korean earth science society
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    • v.38 no.1
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    • pp.24-34
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    • 2017
  • The westerly waves generation is described in the advanced earth science textbook used at high school as follows: as westerly wind approaches and blows over large mountains, the air flow shows wave motions in downwind side, which can be explained by the conservation of potential vorticity. However, there has been no case study showing the phenomena of the mesoscale westerly waves with observational data in the area of small mountains in Korea. And thus the wind speed and time persistency of westerly winds along with the width and length of mountains have never been studied to explain the generation of the westerly waves. As a first step, we assured the westerly waves generated in the downwind side of Sobaek mountains based on surface station wind data nearby. Furthermore, the critical or minimum wind velocity of the westerly wind over Sobaek mountains to generate the downwind wave were derived and calcuated tobe about $0.6m\;s^{-1}$ for Sobaek mountains, which means that the westerly waves could be generated in most cases of westerly blowing over the mountains. Using surface station data and 4-dimensional assimilation data of RDAPS (Regional Data Assimilation and Prediction System) provided by Korea Meteorological Agency, we also analyzed cases of westerly waves occurrence and life cycle in the downwind side of Sobaek mountains for a year of 2014. The westerly waves occurred in meso-${\beta}$ or -${\gamma}$ scales. The westerly waves generated by the mountains disappeared gradually with wind speed decreasing. The occurrence frequency of the vorticity with meso-${\beta}$ scale got to be higher when the stronger westerly wind blew. When we extended the spatial range of the analysis, phenomena of westerly waves were also observed in the downwind side of Yensan mountains in Northeastern China. Our current work will be a study material to help students understand the atmospheric phenomena perturbed by mountains.

Electrochemical Characteristics of Cu3Si as Negative Electrode for Lithium Secondary Batteries at Elevated Temperatures (리튬 이차전지 음극용 Cu3Si의 고온에서의 전기화학적 특성)

  • Kwon, Ji-Y.;Ryu, Ji-Heon;Kim, Jun-Ho;Chae, Oh-B.;Oh, Seung-M.
    • Journal of the Korean Electrochemical Society
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    • v.13 no.2
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    • pp.116-122
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    • 2010
  • A $Cu_3Si$ film electrode is obtained by Si deposition on a Cu foil using DC magnetron sputtering, which is followed by annealing at $800^{\circ}C$ for 10 h. The Si component in $Cu_3Si$ is inactive for lithiation at ambient temperature. The linear sweep thermammetry (LSTA) and galvano-static charge/discharge cycling, however, consistently illustrate that $Cu_3Si$ becomes active for the conversion-type lithiation reaction at elevated temperatures (> $85^{\circ}C$). The $Cu_3Si$ electrode that is short-circuited with Li metal for one week is converted to a mixture of $Li_{21}Si_5$ and metallic Cu, implying that the Li-Si alloy phase generated at 0.0 V (vs. Li/$Li^+$) at the quasi-equilibrium condition is the most Li-rich $Li_{21}Si_5$. However, the lithiation is not extended to this phase in the constant-current charging (transient or dynamic condition). Upon de-lithiation, the metallic Cu and Si react to be restored back to $Cu_3Si$. The $Cu_3Si$ electrode shows a better cycle performance than an amorphous Si electrode at $120^{\circ}C$, which can be ascribed to the favorable roles provided by the Cu component in $Cu_3Si$. The inactive element (Cu) plays as a buffer against the volume change of Si component, which can minimize the electrode failure by suppressing the detachment of Si from the Cu substrate.

Expression of p53 in Human Primary Lung Cancers (인체 폐암종에서 p53의 발현에 관한 연구)

  • Lee, Young-Kyu;Park, Sung-Soo;Shin, Dong-Ho;Lee, Dong-Hoo;Lee, Jung-Hee;Lee, Jung-Dal
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.4
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    • pp.395-403
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    • 1993
  • Background: The cell cycle is composed of a series of steps which can be negatively or positively regulated by various factors. Alteration or inactivation of p53 by mutations, or by its interactions with oncogene products of DNA tumor viruses, can lead to cancer. Mutations of the p53 gene occur frequently in human primary lung cancers and the wild-type p 53 allele is often concomitantly deleted. These suggest that deprivation of suppressive role of the wild-type p53 may ensure tumor cell growth presumable by the mutant p53 gene. Methods: In an attempt to investigate this hypothesis, a mutant p53 gene was immunohistochemically demonstrated in the formalin-fixed paraffin-embedded tissue sections of lung cancers by using a monoclonal antibody p53 (Ab-3 and clone DO7). Results: The expression of p53 (DO7) was found in all four normal lung tissues, four small cell carcinomas, and four non small cell carcinomas in histologic types of lung cancer. In the six normal lung tissues the expressions of p53 (Ab-3) were not found. Contrarily, the expression of p53 (Ab-3) was found in the nuclei of lung cancers among fifteen (46.9%) of thirty-two cases studied. The expression of p53 (Ab-3) was disclosed in three case (37.5%) of eight small cell carcinomas and twelve cases (50.0%) of twenty-four non small cell carcinomas in histologic types of lung cancer. Conclusion: These findings suggest that expression of the mutant p53 is related to the one of events in the pathogenesis of human lung cancer and the role of the other oncogenes might be also related to the development of lung cancers.

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