• Korean Journal of Microbiology
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• v.52 no.4
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• pp.428-436
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• 2016

The emergence of antibiotic-resistant bacteria has been increased and become a public health concern worldwide. Many bacterial infections can be sequentially treated with different types of antibiotics. Thus, this study was designed to evaluate the changes in survival, antibiotic susceptibility, mutant frequency, ${\beta}$-lactamase activity, biofilm formation, and gene expression in Klebsiella pneumoniae after exposure to sequential antibiotic treatments of ciprofloxacin and meropenem. Treatments include control (CON; no addition), 1/2 MIC ciprofloxacin addition (1/2CIP), 2 MIC ciprofloxacin addition (2CIP), initial 1/2 MIC ciprofloxacin addition followed by 1/2 MIC meropenem (8 h-incubation) and 2 MIC ciprofloxacin (16 h-incubation) (1/2CIP-1/2MER-2CIP), initial 1/2 MIC ciprofloxacin addition followed by 1/2 MIC meropenem (8 h-incubation) and 2 MIC meropenem (16 h-incubation) (1/2CIP-1/2MER-2MER), and initial 1/2 MIC ciprofloxacin addition followed by 2 MIC ciprofloxacin(8 h-incubation) and 2 MIC meropenem(16 h-incubation) (1/2CIP-2CIP-2MER). No growth of K. pneumoniae was observed for the 2CIP throughout the incubation period. The numbers of planktonic cells varied with the treatments (7~10 log CFU/ml), while those of biofilm cells were not significantly different among treatments after 24-h incubation, showing approximately 7 log CFU/ml. Among the sequential treatments, the least mutant frequency was observed at the 1/2CIP-1/2MER-2CIP (14%). Compared to the CON, 1/2CIP-2CIP-2MER decreased the sensitivity of K. pneumoniae to piperacillin, cefotaxime, and nalidixic acid. The highest ${\beta}$-lactamase activity was 22 nmol/min/ml for 1/2CIP-1/2MER-2CIP, while the least ${\beta}$-lactamase activity was 6 nmol/min/ml for 1/2CIP-2CIP-2MER. The relative expression levels of multidrug efflux pump-related genes (acrA, acrB, and ramA) were increased more than 2-fold in K. pneumoniae exposed to 1/2CIP-1/2MER-2MER and 1/2CIP-2CIP-2MER. The results suggest that the sequential antibiotic treatments could change the antibiotic resistance profiles in K. pneumoniae.

• Tuberculosis and Respiratory Diseases
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• v.58 no.5
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• pp.473-479
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• 2005

Background : Gefitinib targets the epidermal growth factor receptor r(EGFR), and Gefitinib has antitumor activity in patient with non-small cell lung cancer (NSCLC). However, only 10 to 20 percent of patients show a clinical response to this drug, and the molecular mechanisms underlying patient sensitivity to gefitinib are unknown. PTEN (Phosphatase and tensin homolog deleted on chromosome Ten) plays a role for the modulation of the phosphatidylinositol 3-kinase pathway (PI3K), which is involved in cell proliferation and survival, so that it can inhibit cell cycle progression and induce G1 arrest. Therefore, we analyzed the relationship between PTEN expression and gefitinib's responsiveness in patients having advanced non small cell lung cancer that had progressed after previous chemotherapy. Methods : The expression of PTEN was studied by immunohistochemistry in paraffin-embedded tumor blocks that were obtained from 22 patients who had been treated with gefitinib from JAN, 2001 to AUG. 2004. For the evaluation of the relationships between the PTEN expression, the clinical stage and the basal characteristics, those cases that showed the respective antigen expression in >50% of the tumor cells were considered positive. Results : The positive rate of PTEN staining was 55% of the total of 22 patients. There was a significant relationship between the increased expression of PTEN and the response group (p=0.039). However, there was no significant relationship between the expression of PTEN and other clinicopathologic characteristics. Conclusion: The expression of PTEN in patients with advanced non small cell lung cancer that has progressed after previous chemotherapy may play a role in gefitinib's responsiveness.

• Neonatal Medicine
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• v.17 no.1
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• pp.21-33
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• 2010

Purpose : The increased survival of preterm infants in the neonatal intensive care unit recently has resulted in an increased frequency of bronchopulmonary dysplasia (BPD), especially with atypical forms. However, there have been few studies compairing classic and atypical BPD. The aim of this study was to investigate the differences between these two types of BPD. Methods : Infants with a gestational age less than 32 weeks born at the Seoul National University Hospital and Bundang Seoul National University Hospital from May 2004 to April 2007 were included. The data were categorized in 2 groups, classic and atypical BPD. We determined the incidence of BPD, and compared perinatal factors and postnatal managements between two groups. Results : Among 260 study subjects, 141 (54.2%) infants had BPD. Classic BPD infants were 64 and atypical BPD infants were 77. Comparison of differences between 2 groups, classic BPD infants were associated with respiratory distress syndrome, patent ductus arteriosus, intrauterine growth restriction, more high-frequency ventilator (HFV) use, low 1 and 5 minute Apgar scores. Atypical BPD infants were associated with antenatal steroid use, maternal premature rupture of membrane and chorioamnionitis (CAM). In multivariate analysis, more HFV use was associated with classic BPD. Antenatal steroid use, clinical CAM and histological CAM were associated with atypical BPD. Conclusion : The results of this study showed that antenatal factors (antenatal steroid use, clinical CAM, histological CAM) were associated with atypical BPD and postnatal factors (HFV used more) were associated with classic BPD. Further studies are needed for prevention and treatment of BPD.

• Journal of Life Science
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• v.18 no.4
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• pp.522-529
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• 2008

High-temperature requirement A2(HtrA2) has been known as a human homologue of bacterial HtrA that has a molecular chaperone function. HtrA2 is mitochondrial serine protease that plays a significant role in regulating the apoptosis; however, the physiological function of HtrA2 still remains elusive. To establish experimental system for the investigation of new insights into the function of HtrA2 in mammalian cells, we first obtained $HtrA2^{+/+}$ and $HtrA2^{-/-}$ MEF cells lines and identified those cells based on the expression pattern and subcellular localization of HtrA2, using immunoblot and biochemical assays. Additionally, we observed that the morphological characteristics of $HtrA2^{-/-}$ MEF cells are different form those of $HtrA2^{+/+}$ MEF cells, showing a rounded shape instead of a typical fibroblast-like shape. Growth rate of $HtrA2^{-/-}$ MEF cells was also 1.4-fold higher than that of $HtrA2^{+/+}$ MEF cells at 36 hours. Furthermore, we verified both MEF cell lines induced caspsase-dependent cell death in response to apoptotic stimuli such as heat shock, staurosporine, and rotenone. The relationship between HtrA2 and heat shock-induced cell death is the first demonstration of the research field of HtrA2. Our study suggests that those MEF cell lines are suitable reagents to further investigate the molecular mechanism by which HtrA2 regulates the balance between cell death and survival.

• Radiation Oncology Journal
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• v.19 no.2
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• pp.163-170
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• 2001

Purpose : The measurement of radiation survival using a clonogenic assay, the established standard, can be difficult and time consuming. In this study, We have used the MTT assay, based on the reduction of a tetrazolium salt to a purple formazan precipitate by living cells, as a substitution for clonogenic assay and have examined the optimal condition for performing this assay in determination of radiation sensitivity. Materials and Methods : Four human cancer cell lines - PCI-1, SNU-1066, NCI-H630 and RKO cells have been used. For each cell line, a clonogenic assay and a MTT assay using Premix WST-1 solution, which is one of the tetrazolium salts and does not require washing or solubilization of the precipitate were carried out after irradiation of 0, 2, 4, 6, 8, 10 Gy. For clonogenic assay, cells in $25\;cm^2$ flasks were irradiated after overnight incubation and the resultant colonies containing more than 50 cells were scored after culturing the cells for $10\~14$ days. For MTT assay, the relationship between absorbance and cell number, optimal seeding cell number, and optimal timing of assay was determined. Then, MTT assay was performed when the irradiated cells had regained exponential growth or when the non-irradiated cells had undergone four or more doubling times. Results : There was minimal variation in the values gained from these two methods with the standard deviation generally less than $5\%$, and there were no statistically significant differences between two methods according to t-test in low radiation dose (below 6 Gy). The regression analyses showed high linear correlation with the $R^2$ value of $0.975\~0.992$ between data from the two different methods. The optimal cell numbers for MTT assay were found to be dependent on plating efficiency of used cell line. Less than 300 cells/well were appropriate for cells with high plating efficiency (more than $30\%$). For cells with low plating efficiency (less than $30\%$), 500 cells/well or more were appropriate for assay. The optimal time for MTT assay was after 6 doubling times for the results compatible with those of clonogenic assay, at least after 4 doubling times was required for valid results. In consideration of practical limits of assay (12 days, in this study) cells with doubling time more than 3 days were inappropriate for application. Conclusion : In conclusion, it is found that MTT assay can successfully replace clonogenic assay of tested cancer cell lines after irradiation only if MTT assay was undertaken with optimal assay conditions that included plating efficiency of each cell line and doubling time at least.

• Korean journal of applied entomology
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• v.57 no.3
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• pp.127-135
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• 2018

Climate change can affect variables related to the life cycle of insects, including growth, development, survival, reproduction and distribution. As it encourages alien insects to rapidly spread and settle, climate change is regarded as one of the direct causes of decreased biodiversity because it disturbed ecosystems and reduces the population of native species. Hypera postica caused a great deal of damage in the southern provinces of Korea after it was first identified on Jeju lsland in the 1990s. In recent years, the number of individuals moving to estivation sites has concerned scientists due to the crop damage and national proliferation. In this study, we examine how climate change could affect inhabitation of H. postica. The MaxEnt model was applied to estimate potential distributions of H. postica using future climate change scenarios, namely, representative concentration pathway (RCP) 4.5 and RCP 8.5. As variables of the model, this study used six bio-climates (bio3, bio6, bio10, bio12, bio14, and bio16) in consideration of the ecological characteristics of 66 areas where inhabitation of H. postica was confirmed from 2015 to 2017, and in consideration of the interrelation between prediction variables. The fitness of the model was measured at a considered potentially useful level of 0.765 on average, and the warmest quarter has a high contribution rate of 60-70%. Prediction models (RCP 4.5 and RCP 8.5) results for the year 2050 and 2070 indicated that H. postica habitats are projected to expand across the Korean peninsula due to increasing temperatures.

• Radiation Oncology Journal
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• v.22 no.2
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• pp.145-154
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• 2004

Purpose : To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. Materials and Methods : Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37$^{\circ}C$ with 5$\%$ CO$_{2}$ in air atmosphere. After removal of HS-1200, cells were irradiated with 2$\~$8 Gy X-ray, and then cultured Ii drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16$\mu$M of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40$\mu$M of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken. Results : Treatment of MCF-7 cells with 40$\mu$M of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential ($\Delta$$\psi$$_{m}$) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. Conclusion : We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio In HS-1200 co-treatment group underlies the increased radio sensitivity of MCF-7 cells. Further futures studies are remained elusive.

• Childhood Kidney Diseases
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• v.6 no.2
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• pp.131-141
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• 2002

Purpose : Renal transplantation which allows children normal growth and development and a return to normal life. is now proven to be the best modality for children with ESRD Up to Recently, the number of renal transplantations in Asia has rapidly increased and the outcome has also improved. This investigation was planned to estimate the current status of pediatric renal transplantation in Asia and to find the keys for better improvement of outcome in pediatric renal allograft in Asian countries. Material and methods : The participating countries and institutions for this investigation were China, Hong Kong, India, Indonesia, Japan, Malaysia, Pakistan, Philippines, Singapore, Thailand, Korea, KSPN (Korean Society of Pediatric Nephrology), KONOS (Korean Network for Organ Sharing). Results : Many countries in Asia still do not have a well organized nation wide renal transplantation registration system independently in the pediatric field. So it's very difficult to evaluate the real state of pediatric transplantation among Asian countries. According to the estimation with fragmented data from each countries, in the front running countries of pediatric renal transplantation in Asia, about 40 or more transplants were performed in each country per year and the five year actuarial renal allograft survival was around 80% which is similar to that of western countries. But there were large gaps among the behind groups. Conclusion : Vigorous attempts to perform renal transplantation for children especially younger than 5 years old would be encouraged as well as organ donation from brain dead donor and non heart beating cadaveric donor also should be activated to cope effectively with the shortage of living donor supply. Large number of recent reports shows the favorable outcome of pre-emptive renal transplantation, we should make more efforts toward pre-emptive renal transplantation. First of all, in order to improve the outcome and to narrow the gap between Asian countries in pediatric renal transplantation, effective and continuous efforts to establish nationwide pediatric renal transplantation registration program as well as official, nation-to-nation data sharing program should be needed.

• Journal of Korean Society of Forest Science
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• v.82 no.3
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• pp.221-226
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• 1993

Methods for shoot proliferation via pulse treatment onto the microshoots of Quercus acutissima, and ex vitro root induction using peat plug systems of the microshoots of 4 oak trees were described. Pulsing solution was prepared by the addition of BA and/or BA plus zeatin onto the aqueous WPM and sterilized distilled water. Using the solution, pulsing time was adjusted at different levels(0. 1, 2, 5. 9, and 24 hours). Although the effect of pulsing solution prepared by the addition of cytokinins onto the sterilized distilled water was slightly lower in shoot proliferation rate, a little higher in shoot elongation was observed compared with that of aqueous WPM. One hour of pulse treatment revealed best in shoot proliferation and its elongation, whereas the increment of pulsing time slightly suppressed the response. In addition, prolonged pulse time resulted high frequency of hyperhydric shoot appearance. Single treatment of BA was better in shoot proliferation than that of BA combination with zeatin, whereas the latter treatment usually showed rapid and healthy shoot growth. For ex vitro root induction using peat plug systems, black oaks(Q. acutissima and Q. variabilis) revealed excellent rootability compared with white oaks(Q. serrata and Q. mongolica). Shoot-tip necrosis of white oaks eras one of the big problems for survival. In this study, we discribed the effect of pulse treatment, successful ex vitro rooting system by the incorporation of peat plug, and the possibilities for the overcoming the obstacles on micropropagation of oaks.

• Childhood Kidney Diseases
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• v.11 no.2
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• pp.239-246
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• 2007

Purpose : The progressive deterioration of renal function in children can impose a serious and lifelong impact on their lives. The ultimate goal in the management of children with chronic kidney disease(CKD) is to prolong survival, to prevent complications, and to promote growth and neurodevelopment. The aim of this study is to investigate the risk factors for the decline of renal function in pediatric CKD patients. Methods : Data from patients who met the criteria for the Kidney Disease Outcomes Quality Initiative(K/DOQI) CKD stage 2 to 4 between August 1999 and March 2007 were retrospectively analyzed. The estimated glomerular filtration rate(eGFR) was calculated by the Schwartz formula, using serum creatinine levels and height. We calculated the annual eGFR change from the difference between the baseline eGFR and the last eGFR divided by the duration(years) of the follow-up period. We analyzed the association between the annual eGFR change and factors such as age, gender, K/DOQI stage, underlying renal disease, serum calcium, and inorganic phosphorous during the follow-up period. Results : Sixty one children(44 boys & 17 girls) were enrolled. The age at entry was $7.1{\pm}4.7$ years. The annual eGFR change was $-1.2{\pm}11.9 mL/min/1.73m^2/year$. Our study showed that older age(P=0.005). hypocalcemia(P=0.012), and hypenhosphatemia(P=0.002) were significantly related to more rapid decline in renal function. Conclusion : In pediatric CKD, older age, hypocalcemia and hyperphosphatemia are related to more rapid deterioration of renal function.