• Title/Summary/Keyword: Expression Concentration

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The Mechanism of Interferon-$\gamma$ Induced Cytotoxicity on the Lung Cancer Cell Line, A549 (인터페론감마에 의한 A549 폐암세포주 세포독성의 기전)

  • Oh, Yeon-Mok;Yoo, Chul-Gyu;Chung, Hee-Soon;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.63-68
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    • 1996
  • Background: Interferon-$\gamma$ has various biologic effects, including antiviral effect, antitumor proliferative effect, activation of macrophage and B lymphocyte, and increased expression of major histocompatibility complex. Especially, antitumor proliferative effect of interferon-$\gamma$ has already been proved to be important in vivo as well as in vitro. And, clinical studies of interferon-$\gamma$ have been tried in lung cancer patients. However, the mechanism of antitumor effect of interferon-$\gamma$ has not yet been established despite of many hypotheses. "Necrosis" is a type of cell death which is well known to occur in the circumstances of severe stresses. In contrast, "apoptosis" is another type of cell death which occurs in such biological circumstances as embryonic development, regression of organs, and self-tolerance of lymphocytes. And, apoptosis is an active process of cell death in which cells are dying with fragmentations of their cytoplasms and nuclei. And, in the process of apoptosis the DNAs of cells are cleaved between nucleosomes by unidentified endonuclease and therefore DNAs of apoptotic cells result in a typical electrophoresis pattern known as DNA ladder pattern. Recently it has been suggested that cytotoxic effect of interferon-$\gamma$ occurs via apoptosis. To elucidate the mechanism of antitumor cytotoxic effect of interferon-$\gamma$, we microscopically observed a lung cancer cell line, A549 which was treated with interferon-$\gamma$. We observed A545 treated with interferon-$\gamma$ was dying fragmented. And so, we performed this study to find out that the mechanism of antitumor cytotoxic effect of interferon-$\gamma$ be apoptosis. Method: We treated A549, human lung cancer cell line with various concentration of interferon-$\gamma$ and quantified its cytotoxic effect of various periods, 24 hours, 72 hours and, 120 hours by MTT(dimethylthiazolyl diphenyltetrazolium bromide) bioassay. Also, after we treated A549 with 100 units/mi of interferon-$\gamma$ for 120 hours, we observed the pattern of cell death with inverted microscope and we extracted DNAs from the dead A549 cells and observed the pattern of 1.5% agarose gel electrophoresis with ethidium bromide staining. Result: 1) Cytotoxic effect of interferon-$\gamma$ on A549: For the first 24 hours, threre was little cytotoxic effect and for between 24 hours and 72 hours, there was the beginning of cytotoxic effect and for 120 hours there was increased cytotoxic effect. 2) Pattern of A549 cell death by interferon-$\gamma$: We observed with inverted microscope that A549 cells were dying fragmented. 3) DNA ladder pattern of gel electrophoresis: We observed DNA ladder pattern of gel electrophoresis of extracted DNAs from dead A549 cells. Conclusion: We concluded that the mechanism of interferon-$\gamma$induced cytotoxicity on lung cancer cell line, A549 be via apoptosis.

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Inhibition of Neointima Formation and Migration of Vascular Smooth Muscle Cells by Anti-vascular Endothelial Growth Factor Receptor-1 (Flt-4) Peptide in Diabetic Rats (당뇨병 쥐에서 혈관내피 성장인자 수용체-1 차단 펩타이드를 이용한 신내막 형성과 혈관평활근세포 이동의 억제)

  • Jo, Min-Seop;Yoo, Ki-Dong;Park, Chan-Beom;Cho, Deog-Gon;Cho, Kue-Do;Jin, Ung;Moon, Kun-Woong;Kim, Chul-Min;Wang, Young-Pil;Lee, Sun-Hee
    • Journal of Chest Surgery
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    • v.40 no.4 s.273
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    • pp.264-272
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    • 2007
  • Background: Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis, including stimulating the proliferation and migration of vascular smooth muscle cells (VSMCs). It has been known that diabetes is associated with accelerated cellular proliferation via VEGF, as compared to that under a normal glucose concentration. We investigated the effects of selective blockade of a VEGF receptor by using anti-Flt-1 peptide on the formation and hyperplasia of the neointima in balloon injured-carotid arteries of OLETF rats and also on the in vitro VSMCS' migration under high glucose conditions. Material and Method: The balloon-injury method was employed to induce neointima formation by VEGF. For f4 days beginning 2 days before the ballon injury, placebo or vascular endothelial growth factor receptor-1 (VEGFR-1) specific peptide (anti-Flt-1 peptide), was injected at a dose of 0.5mg/kg daily into the OLETF rats. At 14 days after balloon injury, the neointimal proliferation and vascular luminal stenosis were measured, and cellular proliferation was assessed by counting the proliferative cell nuclear antigen (PCNA) stained cells. To analyze the effect of VEGF and anti-Flt-1 peptide on the migration of VSMCs under a high glucose condition, transwell assay with a matrigel filter was performed. And finally, to determine the underlying mechanism of the effect of anti-Flt-1 peptide on the VEGF-induced VSMC migration in vitro, the expression of matrix metalloproteinase (MMP) was observed by performing reverse transcription-polymerase chain reaction (RT-PCR). Result: Both the neointimal area and luminal stenosis associated with neointimal proliferation were significantly decreased in the anti-Flt-1 peptide injected rats, ($0.15{\pm}0.04 mm^2$ and $ 36.03{\pm}3.78%$ compared to $0.24{\pm}0.03mm^2\;and\;61.85{\pm}5.11%$, respectively, in the placebo-injected rats (p<0.01, respectively). The ratio of PCNA(+) cells to the entire neointimal cells was also significantly decreased from $52.82{\pm}4.20%\;to\;38.11{\pm}6.89%$, by the injected anti-Flt-1 peptide (p<0.05). On the VSMC migration assay, anti-Flt-1 peptide significantly reduced the VEGF-induced VMSC migration by about 40% (p<0.01). Consistent with the effect of anti-Flt-1 peptide on VSMC migration, it also obviously attenuated the induction of the MMP-3 and MMP-9 mRNA expressions via VEGF in the VSMCS. Conclusion: Anti-Flt-1 peptide inhibits the formation and hyperplasia of the neointima in a balloon-injured carotid artery model of OLETF rats. Anti-Flt-1 peptide also inhibits the VSMCs' migration and the expressions of MMP-3 and MMP-9 mRNA induced by VEGF under a high glucose condition. Therefore, these results suggest that specific blockade of VEGFR-1 by anti-Flt-1 peptide may have therapeutic potential against the arterial stenosis of diabetes mellitus patients or that occurring under a high glucose condition.

Derivation of the Synthetic Unit Hydrograph Based on the Watershed Characteristics (유역특성에 의한 합성단위도의 유도에 관한 연구)

  • 서승덕
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.17 no.1
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    • pp.3642-3654
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    • 1975
  • The purpose of this thesis is to derive a unit hydrograph which may be applied to the ungaged watershed area from the relations between directly measurable unitgraph properties such as peak discharge(qp), time to peak discharge (Tp), and lag time (Lg) and watershed characteristics such as river length(L) from the given station to the upstream limits of the watershed area in km, river length from station to centroid of gravity of the watershed area in km (Lca), and main stream slope in meter per km (S). Other procedure based on routing a time-area diagram through catchment storage named Instantaneous Unit Hydrograph(IUH). Dimensionless unitgraph also analysed in brief. The basic data (1969 to 1973) used in these studies are 9 recording level gages and rating curves, 41 rain gages and pluviographs, and 40 observed unitgraphs through the 9 sub watersheds in Nak Oong River basin. The results summarized in these studies are as follows; 1. Time in hour from start of rise to peak rate (Tp) generally occured at the position of 0.3Tb (time base of hydrograph) with some indication of higher values for larger watershed. The base flow is comparelatively higher than the other small watershed area. 2. Te losses from rainfall were divided into initial loss and continuing loss. Initial loss may be defined as that portion of storm rainfall which is intercepted by vegetation, held in deppression storage or infiltrated at a high rate early in the storm and continuing loss is defined as the loss which continues at a constant rate throughout the duration of the storm after the initial loss has been satisfied. Tis continuing loss approximates the nearly constant rate of infiltration (${\Phi}$-index method). The loss rate from this analysis was estimated 50 Per cent to the rainfall excess approximately during the surface runoff occured. 3. Stream slope seems approximate, as is usual, to consider the mainstreamonly, not giving any specific consideration to tributary. It is desirable to develop a single measure of slope that is representative of the who1e stream. The mean slope of channel increment in 1 meter per 200 meters and 1 meter per 1400 meters were defined at Gazang and Jindong respectively. It is considered that the slopes are low slightly in the light of other river studies. Flood concentration rate might slightly be low in the Nak Dong river basin. 4. It found that the watershed lag (Lg, hrs) could be expressed by Lg=0.253 (L.Lca)0.4171 The product L.Lca is a measure of the size and shape of the watershed. For the logarithms, the correlation coefficient for Lg was 0.97 which defined that Lg is closely related with the watershed characteristics, L and Lca. 5. Expression for basin might be expected to take form containing theslope as {{{{ { L}_{g }=0.545 {( { L. { L}_{ca } } over { SQRT {s} } ) }^{0.346 } }}}} For the logarithms, the correlation coefficient for Lg was 0.97 which defined that Lg is closely related with the basin characteristics too. It should be needed to take care of analysis which relating to the mean slopes 6. Peak discharge per unit area of unitgraph for standard duration tr, ㎥/sec/$\textrm{km}^2$, was given by qp=10-0.52-0.0184Lg with a indication of lower values for watershed contrary to the higher lag time. For the logarithms, the correlation coefficient qp was 0.998 which defined high sign ificance. The peak discharge of the unitgraph for an area could therefore be expected to take the from Qp=qp. A(㎥/sec). 7. Using the unitgraph parameter Lg, the base length of the unitgraph, in days, was adopted as {{{{ {T}_{b } =0.73+2.073( { { L}_{g } } over {24 } )}}}} with high significant correlation coefficient, 0.92. The constant of the above equation are fixed by the procedure used to separate base flow from direct runoff. 8. The width W75 of the unitgraph at discharge equal to 75 per cent of the peak discharge, in hours and the width W50 at discharge equal to 50 Per cent of the peak discharge in hours, can be estimated from {{{{ { W}_{75 }= { 1.61} over { { q}_{b } ^{1.05 } } }}}} and {{{{ { W}_{50 }= { 2.5} over { { q}_{b } ^{1.05 } } }}}} respectively. This provides supplementary guide for sketching the unitgraph. 9. Above equations define the three factors necessary to construct the unitgraph for duration tr. For the duration tR, the lag is LgR=Lg+0.2(tR-tr) and this modified lag, LgRis used in qp and Tb It the tr happens to be equal to or close to tR, further assume qpR=qp. 10. Triangular hydrograph is a dimensionless unitgraph prepared from the 40 unitgraphs. The equation is shown as {{{{ { q}_{p } = { K.A.Q} over { { T}_{p } } }}}} or {{{{ { q}_{p } = { 0.21A.Q} over { { T}_{p } } }}}} The constant 0.21 is defined to Nak Dong River basin. 11. The base length of the time-area diagram for the IUH routing is {{{{C=0.9 {( { L. { L}_{ca } } over { SQRT { s} } ) }^{1/3 } }}}}. Correlation coefficient for C was 0.983 which defined a high significance. The base length of the T-AD was set to equal the time from the midpoint of rain fall excess to the point of contraflexure. The constant K, derived in this studies is K=8.32+0.0213 {{{{ { L} over { SQRT { s} } }}}} with correlation coefficient, 0.964. 12. In the light of the results analysed in these studies, average errors in the peak discharge of the Synthetic unitgraph, Triangular unitgraph, and IUH were estimated as 2.2, 7.7 and 6.4 per cent respectively to the peak of observed average unitgraph. Each ordinate of the Synthetic unitgraph was approached closely to the observed one.

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Studies for Antibiotic Free Chicken Production Using Water Extracts from Artemisia capillaris and Camellia sinensis (인진쑥 및 녹차 추출물을 이용한 무항생제 닭고기 생산 연구)

  • Kim, Dong-Wook;Kim, Ji-Hyuk;Kang, Geun-Ho;Kang, Hwan-Ku;Park, Sung-Bok;Park, Jae-Hong;Bang, Han-Tae;Kim, Min-Ji;Na, Jae-Cheon;Chae, Hyun-Suk;Choi, Hee-Chul;Suh, Ok-Suk;Kim, Sang-Ho;Kang, Chang-Won
    • Food Science of Animal Resources
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    • v.30 no.6
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    • pp.975-988
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    • 2010
  • Two experiments were conducted to determine whether water extracts from Artemisia capillaries (A. capillaries) and Camellia sinensis (C. sinensis) could be used as alternatives to antibiotic growth promoters in broiler feed. The experiment 1 was verified their chemical composition, extracts yields, total phenolic compounds concentration, antioxidant activity, antimicrobial activity, and chicken splenocytes proliferation through in vitro test. The extract yields of A. capillaries and C. sinensis were 26.5 and 16.8%, respectively. Total phenolic compounds concentrations of them expressed as gallic acid equivalent were 15.28 and 26.74 mg/mL, respectively. Electron donating abilities of them expressed as $SC_{50}$ showing 50% DPPH radical scavenging were 0.30 and 0.06 mg, respectively. Bacterial inhibitory rates of them against Escherichia coli, Staphylococcus aureus, and Salmonella Typhimurium were ranged from 42.1 to 52.3% and from 21.6 to 33.7%, respectively. And, these extracts increased proliferation of chicken splenocytes. Especially, A. capillaris was more excellent than Echinacea and Concanavalin A known as T-cell stimulator. The experiment 2 was investigated their effects on growth performance, relative organ weight, cecal microflora, blood biochemical parameters, and splenic cytokines mRNA expression in broiler chicks. Four hundred eighty 1-day-old male broiler chicks (Ross 308) were divided in to 4 treatment groups with 4 replicates of 30 birds in each group: NC (control, no antibiotics), PC (avilamycin, 10 ppm; salinomycin, 60 ppm), AC (A. capillaries, 100 ppm), and CS (C. sinensis, 100 ppm); treatments were administered through water supplementation. Final body weight was significantly higher in all treated groups than in NC (p<0.05). Cecal Salmonella numbers were significantly or somewhat decreased in all treated groups than in NC (p<0.05). The relative weights and lengths of the small intestine were more significantly decreased in the PC and AC groups than in the other groups. Cecal Salmonella numbers were significantly or somewhat decreased in all treated groups than in the NC group (p<0.05). The contents of total cholesterol, aspatate aminotransferase, and alanine aminotransferase in blood serum were more significantly decreased in all treated groups than in NC (p<0.05). In conclusion, these results suggested the possibility that these extracts could serve as alternatives for antibiotic growth promoters.