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An Evaluation of Polycross Progenies for Leaf and Plant Characteristics in Winter Active Tall Fescue (Festuca arundinacea Schreb.) - I. Summer Forage Phase (동기생육형(冬期生育型) 톨페스큐의 엽(葉)및 지상부형질(地上部形質)에 관(關)한 다교배(多交配) 후대검정(後代檢定))

  • Kim, Dal Ung
    • Korean Journal of Agricultural Science
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    • v.2 no.2
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    • pp.357-373
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    • 1975
  • This study was conducted to evaluate the winter active polycross progenies of 10 genotypes selected at the hot and dry climate of the Southern Oregon in their performance in the progeny test comparing with a high yielding variety, 'Fawn', and a winter active variety, 'TFM', as the control varieties at Daejon, Korea. Various plant and leaf characteristics, especially which related to photosynthesis, and forage production during the first summer after their establishment, were examined. The important conclusions of this study are summarized as follows: 1. The winter active genotypes and variety had less leaf fresh weight and dry weight per leaf than variety 'Fawn'. Variations among polycross progenies of genotypes for these characteristics were great. 2. The winter active genotypes and variety had less leaf area per leaf than variety 'Fawn'. Leaf area among polycross progenies of genotypes deviated greatly and poly cross progenies of 'genotype-16' had the same average leaf area as 'Fawn'. 3. Differences of specific leaf weight (S. L. W.) in the winter active genotypes and variety were not significant. Probably the genetic diversity for S. L. W were not big and were narrowed down already in this genetic population. It was suggested that the photosynthate production within the population might not be different and there might be differences in the photosynthate production-translocation balance. Further study for the diurnal change in S. L. W. within the population might be useful. 4. The winter active variety and genotypes had less leaf width than 'Fawn' does. Leaf width among polycross progenies of genotypes deviated significantly. 5. Differences among controls and polycross progeny group in the initial plant height were significant and variety 'Fawn' was taller than the winter active genotypes and variety. But the differences were not significant in the regrowth of plant height after the first forage harvest. On the contrary. the differences among polycross progenies of genotypes were not significant in the initial plant but the differences in their polycross progeny performance became obvious and great in the regrowth ability which is an improtent agronomic characteristics for forage crops produced in the pasture and for hay and silage. 6. Plant width of the winter active genotypes and variety was lesser than 'Fawn' variety. 7. Differences of tiller number became evident and variety 'Fawn' had higher tiller number than the winter active genotypes and variety after the first forage cutting. There, deviations among polycross progenies of genotypes were great for this characteristic. It was obvious that the genetic differences became more evident in the second measurement after the first cutting of forage probably because this characteristic were stimulated by defoliation in the cartain genotypes and variety. 8. The winter active genotypes and variety on the initial growth. the regrowth ability andtotal yield had lesser forage yield than variety 'Fawn'. Deviation of forage yield among polycross progenies of genotypes were great and gave basis for selection according to their polycross progeny performance improving the forage yield of these winter active tall fescue population during summer. 9. It was concluded that the winter active variety and genotypes in this study was poorer than variety 'Fawn' for the most of leaf and plant characteristics including forage yield. For these measurements, the variations among polycross progenies of genotypes were great. and plant breeding might able to improve further this winter active tall fescue through the polycross progeny testing method for the higher forage production during summer in Korea. 10. The result of the associations among various characteristics under study were quite agreeable with the results of the analysis of variance and woul be useful in the selection of desirable genotypes for the development of a new variety.

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Diagnosis of the Field-grown Rice Plant -II. Diagnosis by total plant analysis (포장재배(圃場栽培) 수도(水稻)의 영양진단(營養診斷) -II. 전분석(全分析)에 의(依)한 진단(診斷))

  • Park, Hoon;Park, Chon Suh
    • Korean Journal of Soil Science and Fertilizer
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    • v.6 no.3
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    • pp.165-172
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    • 1973
  • The optimum time for nutritional diagnosis of the field-grown rice plant by total plant analysis, and the relationship between maximum or minimum nutrient content at various growth stages and corresponding yield and between maximum or minimum yield and corresponding nutrient content were as follows. 1. The percentage occurence of the minimum nutrient content in straw or grain of minus nutrient plot was in the order of 20 days after transplanting (20)>maximum tillering (MT)>harvested straw (HS)> earformation (EF)>straw at flowering (FS)>harvested grain (HG)>ear at flowering (FE) for nitrogen, MT>EF>HS>20=FS>FE>HG for phosphorus and MT>EF>20>FS>HG>FE for potassium. 2. The time when the occurece of minimum nutrient content in minus plot is highest was considered as the optimum time for nutritional diagnosis of root zone. It was 20 days after transplanting in N and maximum tillering stage in P and K. 3. The highest relative difference($100{\times}(L-H)/H$), between maximum (H)and minimum(L) nutrient content appeared in harvested straw for N and P while in harvested grain for K and Si, suggesting the close relation to their translocation from straw to grain. 4. The corresponding yield of maximum nutrient content was higher than that of minimum content at all growth stages in N, at all stages except MT and EF in P, at 20 days after trans planting and harvest in K, but it was always lower in Si, thus the contribution of nutrient content to yield will be in the order of N>P>K>Si. 5. The highest relative difference ($100{\times}(L-H)/H$, where H and L stand for yields) between yields corresponding to maximum and minimum nutrient content appeared at 20 days after transplanting for N. P. K, indicating the time of the closest relation between yield and nutrient content. 6. The highest difference (H-L, where H and L stand for nutrient content) between N. P. K contents corresponding to maximum or minimum yields came at 20 days after transplanting. The contents of N. P. K corresponding to the maximum total dry matter yield were lower than those corresponding the maximum grain yield at this stage. These facts support the closest relation between yield and nutrient content at this time. 7. The highest yield among yields corresponding to maximum nutrient contents occured at 20 days after transplanting in N. P. K but the lowest yield among yields corresponding to minimum nutrient contents appeared at the same stage only in nitrogen. 8. From the above facts the optimum time for diagnosis of nutrient around root zone seems different from that for diagnosis of nutritional status in relation to grain yield.

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Pro-inflammatory Cytokine Expression Through NF-${\kappa}B/I{\kappa}B$ Pathway in Lung Epithelial Cells (폐 상피세포에서 NF-${\kappa}B/I{\kappa}B$ 경로에 의한 염증매개 사이토카인의 발현)

  • Park, Gye-Young;Lee, Seung-Hee;HwangBo, Bin;Yim, Jae-Joon;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.3
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    • pp.332-342
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    • 2000
  • Background : The importance of pro-inflammatory cytokines, especially tumor necrosis factor $\alpha$ (INF-$\alpha$) and interleukin-1$\beta$ (IL-1$\beta$), have been extensively documented in the generation of inflammatory lung disease. Lung epithelial cells are also actively involved in initiating and maintaining inflammation by producing pro-inflammatory mediators. Understanding the mechanism of pro-inflammatory cytokine expression in lung epithelial cells is crucial to the development of new therapeutic modalities for inflammatory lung disease. Transcription of most pro-inflammatory cytokines is dependent on the activation of NF-${\kappa}B$. However, the relationship between pro-inflammatory cytokine expression and NF-${\kappa}B/I{\kappa}B$ pathway in lung epithelial cells is not clear. Methods : BEAS-2B, A549, Na-H157, NCI-H719 cells were stimulated with IL-$1{\beta}$ or TNF-$\alpha$ at various times, and then IL-8 and TNF-$\alpha$mRNA expressions were assayed by Northern blot analysis. IL-$1{\beta}$ or TNF-$\alpha$-induced NF-${\kappa}B$ activation was assessed by the nuclear translocation of p65 NF-${\kappa}B$ subunit. The degradation of $I{\kappa}B{\alpha}$ and $I{\kappa}B{\beta}$ by IL-$1{\beta}$ or TNF-$\alpha$stimulation was assayed by Western blot analysis. The phosphorylation of $I{\kappa}B{\alpha}$ was evaluated by Western blot analysis after pre-treating cells with proteasome inhibitor followed by IL-$1{\beta}$ or TNF-$\alpha$ stimulation. The basal level of IKK $\alpha$ expression was evaluated by Western blot analysis. Results: $I{\kappa}B{\alpha}$ and $I{\kappa}B{\alpha}$ was rapidly degraded after 5 minutes of incubation with IL-$1{\beta}$ or TNF-$\alpha$ in BEAS-2B, A549, and NCI-H157 cells. The activation of NF-${\kappa}B{\alpha}$ and the induction of IL-8 and TNF-$\alpha$ mRNA expression were observed by IL-$1{\beta}$ or TNF-$\alpha$ stimulation in these cells. In contrast, neither the changes in NF-${\kappa}B/I{\kappa}B$ pathway nor IL-8 and TNF-$\alpha$mRNA expression was induced by IL-$1{\beta}$ or TNF-$\alpha$ stimulation in NCI-H719 cells. IL-$1{\beta}$ and TNF-$\alpha$-induced $I{\kappa}B$ phosphorylation was observed in BEAS-2B, A549, and NCI-H157 cells, but not in NCI-H719 cells. The basal level of IKK$\alpha$ expression was not different between cell. Conclusion : NF-${\kappa}B/I{\kappa}B$ pathway plays an important role in the expression of pro-inflammatory cytokine in most lung epithelial cells. The absence of the effect on NF-${\kappa}B/I{\kappa}B$ pathway in NCI-H719 cells sæms to be due to the defect in the intracellular signal transduction pathway upstream to IKK.

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Expression of Phospholipase C Isozymes in Human Lung Cancer Tissues (인체 폐암조직에서 Phospholipase C 동위효소의 발현양상)

  • Hwang, Sung-Chul;Mah, Kyung-Ae;Choi, So-Yeon;Oh, Yoon-Jung;Choi, Young-In;Kim, Deog-Ki;Lee, Hyung-Noh;Choi, Young-Hwa;Park, Kwang-Ju;Lee, Yi-Hyeong;Lee, Kyi-Beom;Ha, Mahn-Joon;Bae, Yoon-Su
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.3
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    • pp.310-322
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    • 2000
  • Background : Phospholipase C(PLC) plays an important role in cellular signal transduction and is thought to be critical in cellular growth, differentiation and transformation of certain malignancies. Two second messengers produced from the enzymatic action of PLC are diacylglycerol (DAG) and inositol 1, 4, 5-trisphosphate (IP3). These two second messengers are important in down stream signal activation of protein kinase C and intracellular calcium elevation. In addition, functional domains of the PLC isozymes, such as Src homology 2 (SH2) domain, Src homology 3 (SH3) domain, and pleckstrin homology (PH) domain play crucial roles in protein translocation, lipid membrane modificailon and intracellular memrane trafficking which occur during various mitogenic processes. We have previously reported the presence of PLC-${\gamma}1$, ${\gamma}2$, ${\beta}1$, ${\beta}3$, and ${\delta}1$ isozymes in normal human lung tissue and tyrosine-kinase-independent activation of phospholipase C-${\gamma}$ isozymes by tau protein and AHNAK. We had also found that the expression of AHNAK protein was markedly increased in various mstologic types of lung can∞r tissues as compared to the normallungs. However, the report concerning expression of various PLC isozymes in lung canærs and other lung diseases is lacking. Therefore, in this study we examined the expression of PLC isozymes in the paired surgical specimens taken from lung cancer patients. Methods : Surgically resected lung cancer tissue samples taken from thirty seven patients and their paired normal control lungs from the same patients, The expression of various PLC isozymes were studied. Western blot analysis of the tissue extracts for the PLC isozymes and immunohistochemistry was performed on typical samples for localization of the isozyme. Results : In 16 of 18 squamous cell carcinomas, the expression of PLC-${\gamma}1$ was increased. PLC-${\gamma}1$ was also found to be increased in all of 15 adenocarcinoma patients. In most of the non-small cell lung cancer tissues we had examined, expression of PLC-${\delta}1$ was decreased. However, the expression of PLC-${\delta}1$ was markedly increased in 3 adenocarcinomas and 3 squamous carcinomas. Although the numbers were small, in all 4 cases of small cell lung cancer tissues, the expression of PLC-${\delta}1$ was nearly absent. Conclusion : We found increased expression of PLC-${\gamma}1$ isozyme in lung cancer tissues. Results of this study, taken together with our earlier findings of AHNAK protein-a putative PLD-${\gamma}$, activator-over-expression, and the changes observed in PLC-${\delta}1$ in primary human lung cancers may provide a possible insight into the derranged calcium-inositol signaling pathways leading to the lung malignancies.

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Studies on the Flowering and Maturity in Sesame (Sesamum indicum L.) IV. Effects of Foliage Clipping on the Seed Maturity (참깨의 개화.등숙에 관한 연구 IV. 적엽처리가 참깨의 등숙에 미치는 영향)

  • Lee, Jung-Il;Kang, Chul-Whan;Son, Eung-Ryong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.2
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    • pp.165-173
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    • 1985
  • The objectives of the study were to investigate the effects of foliage clipping on photosynthesis and grain filling for branch and non branch types under the polyethylene film mulch and non mulch conditions in mono cropping and second cropping after barley in sesame (Sesamum indicum L.), and to improve poor grain filling at later flowering time utilizing these data. One thousand grain weight was more decreased in branch type than in non branch type, in polyethylene film mulch condition than in non mulch condition, and in second cropping after barley than in mono cropping by clipping lower part foliage. Twentyfive percent clipping of lower part foliage showed a little increase than no clipping. Matured grain rate also showed same tendency between branch and non branch type and between mono cropping and second cropping after barley as well as 1,000 grain weight except for polyethylene film mulch. Matured grain rate of 25% foliage clipping at 30 days after flowering in non branch type presented a little increase but decreased in branch type. Clipping of higher part leaves were so serious decrease of matured grain rate that higher part leaves at late maturing time have a major role in photosynthesis. Matured grain rate of foliage clipping at 10 days after flowering was decreased in all treatments. Chlorophyll content of higher part leaves at 50% lower part foliage clipping presented 39% increase compared to same positioned leaves of non treatment, and 66% increase by 50% higher part foliage clipping in lower part leaves. Photosynthetic activity was 58% more increased in 50% lower part foliage clipping than no clipping, but seriously decreased in 50% higher part foliage clipping. Therfore, photosynthates of remained lower part leaves could not only support their own demands, but also any contribution to translocation of photosynthates from source to sink at late maturing time. Harvest index was 28% increased in 25% lower part foliage clipping and 13% decreased in 50% higher part foliage clipping compared to no clipping. Leaf area was 48% increased in 50% lower part foliage clipping compared to the same positioned leaves of no clipping, and only 5% increased in higher part foliage clipping. Productivity by foliage clipping compared to non treatment, was highly decreased in branch type than in non branch type, in second cropping after barley than in mono cropping. Little difference was detected between polyethylene film mulch and non mulch conditions. Twenty five percentage of lower part foliage clipping on mono cropping of non branch type appeared 5% and 8% yield increase in each of polyethylene film mulch and non mulch conditions compared to no clipping, and all decreased in other treatments. Mean loss of productivity by foliage clipping at 10 days after flowering was serious than clipping at 30 days after flowering. As the result, contribution to photosynthesis of source at 10 days after flowering are larger than that at 30 days after flowering in sesame. Fifty percent lower part foliage clipping at 10 days after flowering showed so the most serious yield decrease that lower part leaves at that time were considered as the main role leaves for photosynthesis.

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Studies on the Biological Effects of Chemosterilant, Hempa, on the Rice Weevil (Sitophilus oryzae 1.) and Transmission of Sterility (화학적 불임유기물질 Hempa가 쌀바구미(Sitophilus oryzae L.)에 미치는 생물학적 영향 및 불임성의 전달에 관한 연구)

  • Shim Jai Wook
    • Korean journal of applied entomology
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    • v.12 no.1
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    • pp.1-21
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    • 1973
  • Some experiments were conducted to investigate the effects of the chemosterilant, hempa, on the biology of the rice weevil, Sitophilus oryzae L., and the transmission of the lethal factors in the progeny. One to three days old adult males were fed on the wheat grains treated with concentrations of 0.0625, 0.125, 0.25, and $0.5\%$ of hempa water solution. The effects of the treatment on the mortality, longevity, and the performance of oviposition were examined for the Pl generation, and the hatchability and mortality in the postembryonic development were also tested in the $F_1,\;F_2,\;BC_1,\;F_3,\;and\;BC_2$ generations to analyze the inheritance of the lethal factors. The results obtained were summarized as follows. (1) The average longevity of the treated males were ranged from 26.6 to 30.4 days, and indicated no statistical differences. (2) The mortality of the treated males were ranged between $3.3\%\;and\;13.3\%$ and showed no statistical significance. (3) The overall mean number of eggs laid by a female mated to a treated male with concentrations of 0.0625, 0.125, 0.26 and $0.5\%$ were 3.78, 4.05, 3.75 and 3.61 for the respective treatments, and they were not differ significantly from those of control which were 3.60 per female per 3 day period. The unmated female laid 1.91 in the same period, and significantly differ from those in other experimental groups. (4) The overall mean hatchability of the eggs laid by the females mated with males that had been treated with various concentrations of hempa were 86.82, 64.77, 53.47, 40.33 and $24.78\%$ for the respective concentrations of 0, 0.0625, 0.125, 0.25 and $0.5\%$. The hatchability decreased with the increasing concentrations. (5) The minimum hatchabilities were obtained from the eggs laid in the period of 10-12 days after treatment, then the hatchability increased showing some recovery. The recovery seemed to be very much delayed for the males which had been treated with the greater concentrations. Such a difference in hatchability might be related with the sensitivity of the developmental stages of the sperms, and broader spectrum in the stages and severer effects seemed to be associated with the increased concentrations. (6) The overall mean of larval mortality in the $F_l$ generation were 6.55, 17.89, 27.40, 35.42 and $52.17\%$ for the respective concentrations of 0,0.0625, 0.125,0.25 and $0.5\%$. And there was a tendency to increase in the mortality with the increase of concentrations. (7) The correlation coefficients between per cent sterile eggs and larval mortality for the experimental plots of 0.125, 0.25 and $0.5\%$ treatments showed r=+0.83 and +0.85, respectively, and it seemed to be close correlation between the lethal effects on the embryonic and post-embryonic developments. (8) Since the $SC_{50}$ of the sterile eggs was $0.133\%$ and $SC_{50}$ of the larval mortality was $0.565\%$, it was considered that tile lethal factors expressed more in the egg stages than the larval stages. (9) The ratio of female to male in the $F_l$ adults showed 100 : 125, 100 : 108 and 100 : 124 for the plots of 0.125, 0.25 and $0.5\%$ treatments, respectively. And it n·as considered that the sex ratio distortions might occur with the higher concentrations. (10) When the F, males originated 1.on the eggs had been laid by p, in the period of 16-18 days after treatment, were crossed to normal females $(BC_1)$ and made sib matings $(F_2)$, the per cent sterile eggs of the $BC_1$ generation were 13.88 and $33.04\%$ , and were 31.01 and $38.73\%$ for the $F_2$generation with the plots of 0.0625 and $0.125\%$ treatment, respectively. And these seemed to be a results of the $F_1$ individuals are carrying some chromosomal aberrations (11) The larval mortality was the highest in the $F_2$ plot and followed the female backcross plot, and the least in the male backcrosses. (12) The proportions of 1st and 2nd instar larvae among the larval development at tile 17th day after oviposition were 10.98, 27.26, 32.98 and $15.73\%$ in the normal female $\times$ normal male, $F_1$ female$\times$normal male, normal $female \;\times F_1$ male and $female \;\times F_1$ male plots, respectively. It was considered that the larval development might be delayed by the treatment in the 2nd generation. (13) Per cent larval mortality and sterile eggs were greater in the $F_2$ sib mating plots $(F_3)$ than both of $F_2$ backcrosses. Therefore, it seemed that some of the recessive lethal mutations might affect in the further generations. (14) The sterility, induced by the treatment of chemosterilant, hempa, was considered as the result of the dominant lethal mutations due to chromosomal aberrations such as translocation and/or deletion. The effects of these lethal factors seemed to be inherited tip to 3rd generation after treatment.

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Activation of NF-${\kappa}B$ in Lung Cancer Cell Lines in Basal and TNF-${\alpha}$ Stimulated States (폐암 세포에서 기저 상태와 TNF-${\alpha}$ 자극 시 NF-${\kappa}B$의 활성화)

  • HwangBo, Bin;Lee, Seung-Hee;Lee, Choon-Taek;Yoo, Chul-Gyu;Han, Sung-Koo;Shim, Young-Soo;Kim, Young-Whan
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.5
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    • pp.485-496
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    • 2002
  • Background : The NF-${\kappa}B$ transcription factors control various biological processes including the immune response, acute phase reaction and cell cycle regulation. NF-${\kappa}B$ complexes are retained in the cytoplasm in the basal state and various stimuli cause a translocation of the NF-${\kappa}B$ complexes into the nucleus where they bind to the ${\kappa}B$ elements and regulate the transcription of the target genes. Recent reports also suggest that NF-${\kappa}B$ proteins are involved in oncogenesis, tumor growth and metastasis. High expression of NF-${\kappa}B$ expression was reported in many cancer cell lines and tissues. The constitutive activation of NF-${\kappa}B$ was also reported in several cancer cell lines supporting its role in cancer development and survival. The anti-apoptotic action of NF-${\kappa}B$ is important for cancer survival. NF-${\kappa}B$ also controls the expression of several proteins that are important for cellular adhesion (ICAM-1, VCAM-1) suggesting a role in cancer metastasis. In lung cancer, high expression levels of the NF-${\kappa}B$ subunit p50 and c-Rel were reported. In fact, high expression does not mean a high activity, and the activation pattern of NF-${\kappa}B$ in lung cancer has not been reported. Materials and Methods : In this study, the NF-${\kappa}B$ nuclear binding activity in the basal and TNF-${\alpha}$ stimulated states were exmined in various lung cancer cell lines and compared with the normal bronchial epithelial cell line. Twelve lung cancer cell lines including the non-small cell and small cell lung cancer cell lines (A549, NCI-H358, NCI-H441, NCI-H552, NCI-H2009, NCI-H460, NCI-H1229, NCI-H1703, NCI-H157, NCI-H187, NCI-H417, NCI-H526) and BEAS-2B bronchial epithelial cell line were used. To evaluate the NF-${\kappa}B$ expression and DNA binding activity, western blot analysis and an electrophoretic mobility shift assay with the nuclear protein extracts. Results : The basal expressions of the p65 and p50 subunits were observed in the BEAS-2B cell line and all lung cancer cell lines except for NCI-H358 and NCI-H460. The expression levels of p65 and p50 were increased 30 minutes after stimulation with TNF-${\alpha}$ in BEAS-2B and in 10 lung cancer cell lines. In the NCI-H358 and NCI-H460 cell lines, p65 expression was not observed in the basal and stimulated states and the two p50 related protein levels were higher after stimulation with TNF-${\alpha}$ These new proteins were smaller than p50 and are thought to be variants of p50. In the basal state, NF-${\kappa}B$ was nearly activated in the BEAS-2B and all lung cancer cell lines. The DNA binding activity of the NF-${\kappa}B$ complexes was markedly higher after stimulation with TNF-${\alpha}$ In the BEAS-2B and all lung cancer cell line except for NCI-H358 and NCI-H460, the activated NF-${\kappa}B$ complex was a p65/p50 heterodimer. In the NCI-H358 and NCI-H460 lung cancer cell lines, the NF-${\kappa}B$ complex was variant of a p50/p50 homodimer. Conclusion : The NF-${\kappa}B$ activation pattern in the lung cancer cell lines and the normal bronchial epithelial cell lines was similar except for the activation of a variant of the p50/p50 homodimer in some lung cancer cell linse.

The Study of anti-inflammatory Mechanism with Cobra Venom on Astrocytes of Rats (뇌(腦) 성상세포(星狀細胞)를 대상으로 한 Cobrotoxin의 염증(炎症) 치료(治療) 기전(機轉) 연구(硏究))

  • Yoo, Jae-ryong;Song, Ho-sueb
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.155-167
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    • 2005
  • Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Cobrotoxin on binding affinity of cobrotoxin with P50, $IKK{\alpa}$ and $IKK{\beta}$, activities of NF-${\kappa}B$, Cell viability of astrocyte, expressions of protein molecules of NF-${\kappa}B$ such as P50, P-$1{kappa}B$, $1{\kappa}B$ and iflammation related genes such as Cox-2, iNOS, cPLA2 in the SNP or LPS induced Inflammatory pathway of Rats' astrocytes. Methods : In this study, The expression of cytosolic phospholipase A2, Nitric oxcide, Cyclooxygenase-2 and inducible nitrogen oxide synthase was determined by western blotting with corresponding antibodies, and the generation of NF-${\kappa}B$ was assayed by EMSA method in astrocytes of rats. The Cell viability of astrocytes was determined by MTT assay, and Binding affinity of Cobrotoxin with P50, $IKK{\alpha}$ and $IKK{\beta}$ was assayed by Surface plasmon resonance analysis, and NF-${\kappa}B$ dependent luciferase activity was determined by luciferase analysis, and Uptake of cobrotoxin in astrocytes was identified by Confocal laser scanning microscope Results : 1. Compared with control, LPS-induced NF-${\kappa}B$ DNA binding activity was decreased significantly by 0.1, $0.5{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 2. Compared with control, LPS-induced NF-kB dependent luciferase expression was decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 3. Compared with control, SNP induced P50, $I{\kappa}B$ expressions in astrocyte were decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin and P-$1{\kappa}B$ expression was decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 4. Compared with control, LPS induced P50, $1{\kappa}B$ expressions in astrocyte were decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 5. Compared with control, SNP induced Cox-2, iNOS, CPLA2 expressions in astrocyte were decreased significantly by $1{\mu}g/m{\ell}$ of Cobrotoxin. 6. Compared with control, LPS induced Cox-2, cPLA2 expressions in astrocyte were decreased significantly by 0.1, 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin and iNOS expression was decreased significantly by 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin. 7. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, SNP-induced NF-${\kappa}B$ DNA bindins activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM. 8. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, LPS-induced NF-${\kappa}B$ DNA binding activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM, Cobrotoxin $0.5{\mu}g/m{\ell}$with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM 9. Compared with $0.1{\mu}g/m{\ell}$ of cobrotoxin, SNP induced P50 expressions in astrocyte were increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM. 10. The uptake of the labeled cobrotoxin into the cells was shown under a confocal laser scanning microscope. cobrotoxin was uptaken into the membrane and nucleus of astrocytes. Conclusions : In summary, the present results demonstrate that cobrotoxin directly binds to sulfhydryl group of p50 and IKKS resulting In the reduction of translocation of p50 and IkB release, thereby inhibits activation of NF-${\kappa}B$, and suggest that pico to nanomolar range of cobrotoxin could inhibit the expression of genes in the NF-${\kappa}B$ signal pathway.

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