• Korean Journal of Medicinal Crop Science
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• v.8 no.4
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• pp.334-341
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• 2000

This study was conducted to determine the effects and optimum concentrantion of chemical mutagens, colchicine, EMS (ethyl methan sulfonate), MNU (1-methyl-3-1­-nitrosoguanidinenitro), sodium azide $(NaN_3)$ for induction of mutant plants. In order to induce the mutants of Dianthus superbus L, immature seed were pre-soaked in the warter adding each mutagens and concentration of EMS, colchicine, MNU, and sodium azide $(NaN_3)$. Comparision of morphological characteristic and seed germination in each mutant plants differed depending on mutagen sources and their concentrations. When 0.2% EMS were treated on seed, germination decreased to 12% while untreated control was germinated 76.6% for twenty days. Treatments of colchicine appeared higher germination than other mutagen but not survived. The survival rate was extremely decreased in MNU treatment at 0.5mM and chlorophyll-mutant plantlets were obtained by sodium azide treatment at 0.2mM. Chlorophyll mutants were produced by pre-soaking the immature seed of Dianthus superbus L. with mutagen, sodium azide. The control plants appeared normal green leaf color, while mutant plant after mutagenic treatment of immature seed results in yellow­-green stripes and albino in normal green leaf tissue. RAPD was carried out to check the genetic modification of regenerated plants by mutagen treatments at 0.2mM sodium azide. Three polymorphic DNA fragments out of thirty-seven obtained by RAPDs were observed in regenerated plants using five decamer primers.

• KSBB Journal
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• v.21 no.3
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• pp.204-211
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• 2006

For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

• Tuberculosis and Respiratory Diseases
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• v.54 no.5
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• pp.495-509
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• 2003

Background : Neutrophil-mediated inflammation is usually self-limiting, because neutrophils have a remarkably short life span. Prolonged neutrophil survival, which is caused by decreased spontaneous apoptosis, leads to persistent inflammation in sepsis. Because many inflammatory cytokines, which generate signals that delay apoptosis, are regulated by nuclear factor-${\kappa}B$ transcription factor, we hypothesized that nuclear factor-${\kappa}B$ might be related to the reduced neutrophil apoptosis observed in sepsis. Methods : Neutrophils of healthy volunteers and sepsis patients were freshly isolated from venous blood. Neutrophil apoptosis was assayed with two approaches : by counting apoptotic cells under a microscope and by flow cytometry using Annexin V. The activity of nuclear factor-${\kappa}B$ was assessed by immunofluorescent staining or electrophoretic mobility shift assay. Expression of X-linked inhibitor of apoptosis was measured by western blot assay. Results : We confirmed reduced spontaneous neutrophil apoptosis in patients with sepsis. The number of apoptotic neutrophils in patients with sepsis increased to the level of that in healthy controls after cycloheximide treatment, suggesting that decreased spontaneous neutrophil apoptosis is dependent on de novo protein synthesis. In patients with sepsis, basal neutrophil nuclear factor-${\kappa}B$ was activated compared to the level in healthy controls. Moreover, a blockade of nuclear factor-${\kappa}B$ activity reversed the decreased spontaneous neutrophil apoptosis in sepsis patients. Meanwhile, X-linked inhibition of apoptosis expression, which is regulated by nuclear factor-${\kappa}B$, decreased 24 hours after incubation in healthy persons, but persisted for 24 hours in patients with sepsis. Conclusion : These observations suggest that the reduced spontaneous neutrophil apoptosis observed in patients with sepsis may be related to the induction of survival protein by nuclear factor-${\kappa}B$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.1
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• pp.42-47
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• 2006

L-Ascorbic acid (AsA) is an essential nutrient for prevention of scurvy in humans, primates and guinea pigs that lack $L-gulono-\gamma-lactone$ oxidase which is required for the final step of AsA biosynthesis. AsA participates in various hydroxylation reactions involved in the biosynthesis of collagen. The purpose of this study is to clarify the role of AsA on collagen synthesis in 3T6 fibroblasts and primary cultured cells of chondrocytes. Cells were cultured in medium supplemented with catalase and AsA at various concentration. Supplement of AsA induced collagen synthesis in 3T6 fibroblasts and primary cultured cells of chondrocytes. The most remarkable induction of collagen synthesis by AsA was found in primary cultured chondrocytes. The content of collagen representing the amounts of extracellular matrix significantly increased in the cells of which growth was stimulated by AsA, while it decreased with increasing passage numbers of subculture in cells. It showed that the content of collagen decreased in the medium which contained AsA at the concentration higher than 5.0 mM. However, the contents of collagen to DNA were not different among various AsA concentrations. Supplementing with AsA resulted in enhancement of collagen formation and extracellular matrix. Therefore, there might be a Positive correlation between the activity of catalase and the AsA concentration. Moreover, it can be assumed that AsA stimulates the collagen synthesis by optimizing the cell-culture environment.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.11
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• pp.1589-1594
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• 2016

Piperine [(E,E)-5-(3,4-methtylenedioxyphenyl)-2,4-pentadienolypiperidide] is a principal of Piperaceae, including Piper nigrum L. and Piper longum Linn., which has been used as a spice and traditional medicine. In this study, we investigated whether or not piperine has anti-cancer effects on AGS human gastric cancer cells. The results demonstrated that piperine not only inhibited proliferation using MTT assay but also induced apoptotic bodies using DAPI assay in a dose-dependent manner in response to piperine. Expression levels of p53, Bax (pro-apoptotic), cleaved caspase-9, and cleaved-PARP increased, whereas expression levels of Bcl-2, XIAP (anti-apoptotic), and Akt decreased in a dose-dependent manner compared with the control by western blotting analysis. To identify the connection between phospo-Akt and Bcl-2 family in response to piperine, LY249002 (Akt inhibitor) was treated with piperine ($150{\mu}M$). The results were shown that expression of phospo-Akt was reduced whereas expression of Bax and cleaved-PARP increased in a dose-dependent manner. These results indicate that piperine induced apoptosis in AGS cells and may serve as a chemopreventive or therapeutic agent for human gastric cancer.

• The Korean Journal of Pesticide Science
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• v.5 no.2
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• pp.45-50
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• 2001

In recent, non virus-induced mosaic symptoms(NVMS) on potato leaves were observed in the seed potato fields, and its incidence rate was $5{\sim}20%$ nationwide. It made difficult to rogue out virus-infected plants, and caused much arguments between seed potato production farmers and seed potato inspectors. The objectives of these experiments were to find out the causes of NVMS, and also to induce mosaic symptom(phytotoxicity) on potato plants by treatment of several herbicides. No significant correlations were found between incidence rates of NVMS and values from soil analyses; soil pH, soil EC, organic matter content, and contents of inorganic constituents($P_2O_5,\;NO_3$, Ca, Mg, K) in the soil around the potato planted. The examinations by ELISA, virus indicator plants, and TEM showed that NVMS on potato leaves was not caused by the viruses infection. But, the use of herbicides could induced the NVMS on potato leaves. The incidence rates of potato treated with pendimethalin linuron of 400 mL/10 a, pendimethalin of 200 mL/10 a, pendimethalin.oxadiazon of 300 mL/10 a, and control were 61.1%, 47.2%, 19.4%, and 1.4%, respectively. Based on these results, we confirmed that the treatment of pendimethalin alone and in mixture with other herbicides were the reason of NVMS on potato leaves. The yields among test plots were similar except dicamba treated plot, which decreased by about 23% compared to control plot. When their progenies harvested in 1999 were planted in the following season, no symptoms of mosaic were observed.

• Journal of Chest Surgery
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• v.31 no.5
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• pp.502-508
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• 1998

Proinflammatory cytokines such as tumor necrosis factor-$\alpha$(TNF-$\alpha$) have been implicated in myocardial and organ dysfunction associated with postperfusion syndrome. We tested the hypothesis that cytokine productions are depressed by preoperative cortiosteroid injection for cardiopulmonary bypass(CPB) and the postoperative courses will be better than without steriod pretreated cases. Cardiac surgery was performed in randomized blind fashion for 20 patients from June 1996 to September 1996. In the steroid group(n=10), corticosteroid(dexamethasone 1 mg/kg) was injected 1 hour before anesthetic induction, but in the control group(n=10), nothing was injected. Each of groups were sampled 11 times as scheduled for TNF-$\alpha$ bioassays. We have checked EKG, cardiac enzymes(CPK, LDH with isoenzyme), WBC count preoperative day, one day and three days after operation. Viatal signs were continuously monitored for three postoperaive days. In the postoperative period three patients in the control group had elevated body temperature and four patients had hypotension that required considerable intravenous fluid administration. But steroid injected patients showed normal body temperture and acceptable blood pressures without supportive treatment. CPK enzymes rose in control group higher than steroid group at postoperative 1st and 3rd day(CPK; 1122$\pm$465 vs 567$\pm$271, 864$\pm$42 vs 325$\pm$87), and CPK-MB enzymes rose in control group higher than steroid group at postoperative 1st day(106.4$\pm$115.1 vs 29.5$\pm$22.4)(P=0.02). Arterial tumor necrosis factor-$\alpha$ rose during cardiopulmonary bypass, peaking at 5 minutes before the end of aortic cross clamping(ACC-5min) in steroid group(11.9$\pm$4.7 pg/ml), and 5 minutes before the end of cardiopulmonary bypass(CPB-5min) in control group(22.3$\pm$6.8 pg/ml). The steroid pretreated patients had a shorter period of time in respirator suport time, ICU stay day, hospital admission day. We conclude that corticosteroid suppress cytokine production during and after cardiopulmonary bypass, and may improve the postoperative course through inhibition of reperfusion injury such as myocardial stunning and hemodynamic instability.

• Radiation Oncology Journal
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• v.20 no.3
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• pp.206-214
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• 2002

Purpose : The aim of this study was to assess the effectiveness, survival rate and complications of radiation therapy and chemoradiation treatment in hypopharyngeal cancer. Methods and Materials : From January 1984 to December 1999, 56 patients who had hypopharyngeal carcinoma treated with curative radiation therapy were retrospectively studied. Twenty four patients $(42.9\%)$ were treated with radiation therapy alone (Group I) and $32\;(57.1\%)$ treated with a combination of chemotherapy and radiation (Group II). Total radiation dose ranged from 40.5 to 83. 5 Gy (median 67.9 Gy). Radiotherapy was given with conventional technique in 9 patients $(16.4\%)$, with hyperfractionation I ($1.15\~1.2$ Gy/fr., BID) in 26 $(47.2\%)$, hyperfractionation II (1.35 Gy/fr., BID) in 18 $(32.7\%)$, and accelerated fractionation (1.6 Gy/fr., BID) in 2 $(3.6\%)$. In chemotherapy, 5-FU ($1,000\;mg/m^2$ daily for 5 consecutive days) and cisplatin ($100\;mg/m^2$ on day 1) were administered in a cycle of 3 weeks interval, and a total of 1 to 3 cycles (average 2..3 cycles) were given prior to radiation therapy. Follow up duration was $1\~195$ months (median 28 months). Results : Overall 2 and 5 year survival rates were $40.6\%\;and\;27.6\%;\;50.0\%\;and\;30.0\%$ in Group I, and $36.4\%\;and\;26.3\%$ in Group II, respectively. Complete local control rates in Group I and II were $70.0\%\;and\;67.7\%$, respectively. The response to radiotherapy and nodal stage were statistically significant prognostic factors. The complication rate was increased in Group II and was decreased in hyperfractionation. Conclusion : The response to radiotherapy and nodal stage were valid factors to indicate the degree of control over the hypopharyngeal cancer. The induction cisplatin, 5-Fu chemotherapy was not valid in terms of local control rate and survival rate, but did contribute to an increased complication rate. The use of hyperfractionation was valid to reduce the late radiation complications.

• Journal of Wetlands Research
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• v.8 no.2
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• pp.53-64
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• 2006

A study on the restoration process of a stream ecosystem and the water quality renovation technique by removing algae, vegetation and fish monitoring as evaluating the removal of the algae by dietetic characteristics of fishes were performed on Sangdong stream in the B city after stream restoration it to the artificial stream as the cases, restoring urban stream into close-to-nature stream are being increased domestically with the aim of ecological city. As a result, restoration and rehabilitation of the fundamental stream ecosystem was well maintained 4 years later the reclamation at the moment and total 93 diagnosis which were all vascular plant phylum including 44 families, 73 genuses, 79 species and 14 varieties in flora and vegetation community were observed. 3 families, 8 species and 354 populations in total among Fishes were found and Pseudorasbora Parva, Cyprinus Carpic and Carassius Auratus strongly resistant to water pollution were dominantly appeared in order of 50.5% of Pseudorasbora Parva 21.2% of Cyprinus Carpic, 20.9% of Carassius Auratus, 7.1% of Macropodus chinensis and 0.3% of Misqurnus anguillicaudatus according to relative richness index. It turned out to be that Cyprinus Carpic ingests algae over 90% and Carassius Auratus takes it over 30% according to the analysis about the alimentary object of the fishes as a consequences of algae's excrescent from characteristics of the tested experimental stream. It is reported that a Cyprinus Carpic, about 34 cm in length, ingested wet-weight 43.2g algae on the rough analysis toward the sample which makes us recognize how effective a macro community Cyprinus Carpic is for removing algae.As a consequence of this research, the effect of stream ecosystem characteristics and water quality purification could not be expected by aquatic plants and trees which were eliminated at experimental stream. From now on, a close-to-nature stream should be formed of ecological hydraulic and hydrologic engineered modeling from the beginning so that it can perform the water quality purifying function. It is determined that the structure of food chain will be abundantly influenced by the induction of oversized macro community like Cyprinus Carpic because a biomass of a consumer of higher order is increased. It is estimated that the removal algae by fishes is not effective despite in some cases of dietetic characteristics so much more studies should be executed in the future.

• Radiation Oncology Journal
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• v.16 no.4
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• pp.377-388
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• 1998

Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.