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Comparative Evaluation of Colon Cancer Stemness and Chemoresistance in Optimally Constituted HCT-8 cell-based Spheroids (적정 구성 배양 HCT-8 기반 대장암 스페로이드의 암 줄기세포능 및 항암제 내성 평가의 비교 평가 연구)

  • Lee, Seung Joon;Kim, Hyoung-Kab;Lee, Hyang Burm;Moon, Yuseok
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1313-1319
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    • 2016
  • Cancer is a complex disease heterogeneously composed of various types of cells including cancer stem-like cells responsible for relapse and chemoresistance in the tumor microenvironment. The conventional two-dimensional cell culture-based platform has critical limitations for representing the heterogeneity of cancer cells in the three-dimensional tumor niche in vivo. To overcome this insufficiency, three-dimensional cell culture methods in a scaffold-dependent or -free physical environment have been developed. In this study, we improved and simplified the HCT-8 colon cancer cell-based spheroid culture protocol and evaluated the relationship between cancer stemness and responses of chemosensitivity to 5- Fluorouracil (5-FU), a representative anticancer agent against colon cancer. Supplementation with defined growth factors in the medium and the culture dish of the regular surface with low attachment were required for the formation of constant-sized spheroids containing $CD44^+$ and $CD133^+$ colon cancer stem cells. The chemo-sensitivities of $CD44^+$ cancer stem cells in the spheroids were much lower than those of $CD44^-$ non-stem-like cancer cells, indicating that the chemoresistance to 5-FU is due to the stemness of colon cancer cells. Taken together, the inflammation and oncogenic gut environment-sensitive HCT-8 cell-based colon cancer spheroid culture and comparative evaluation using the simplified model would be an efficient and applicable way to estimate colon cancer stemness and pharmaceutical response to anticancer drugs in the realistic tumor niche.

A Case Study - IT Outsourcing of the Korea Development Bank (산업은행: 금융 IT 아웃소싱 - 공동협력으로 안전한 문을 연다)

  • Kang, Ju-Young;Lee, Jae-Kyu
    • Information Systems Review
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    • v.7 no.2
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    • pp.229-255
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    • 2005
  • The Korea Development Bank promoted a total outsourcing for IT operation in 1999 for the first time in the banking industry. The Korea Development Bank became the center of public attention because the most banks were unwilling to take an outsourcing with external sources for the reason of financial operation accidents, securities, and threats of strikes. After the introduction of the total IT outsourcing, the Korea Development Bank has continuously diagnosed the problems of the IT outsourcing and adopted various proper complements for the enhancement of the IT outsourcing. As the result of the enhancement, the IT outsourcing of the Korea Development Bank marched into the joint liability operation period after going through the outsourcing operation period and the co-operation period. The joint liability operation which is the most leading outsourcing system which is adopted by the Korea Development Bank for the first time in the banking industry. Through the joint liability operation, the Korea Development Bank could accept the most up-to-date IT, concentrate internal manpower on the core capability, and secure flexibility of manpower. Also, the bank changed the relationship between the bank and the external sources from the one-sided relationship between a producers and a consumer to the joint liability relationship on which both sides are responsible for the operation, and could integrate the internal capacity with the professional know-how of the external IT outsourcing company. In this paper, we testified the soundness and validity for the worries of banks about the total IT outsourcing with external sources. And, we arranged the advantages and outcomes of the total IT outsourcing with external sources compared to the IT outsourcing with internal sources. Moreover, we expect that we can improve the closed financial IT outsourcing industry structure and raise the world competitive power of domestic IT outsourcing companies by correcting wrong ideas on the IT outsourcing with external sources.

Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Relationship between Monounsaturated Fatty Acid Composition and Stearoyl-CoA Desaturase mRNA Level in Hanwoo Liver and Loin Muscle (한우 간 및 등심 조직에서 불포화지방산의 조성비율과 Stearoyl-CoA Desaturase mRNA의 발현 양상)

  • Lee, S.H.;Yoon, D.H.;Hwang, S.H.;Cheong, E.Y.;Kim, O.H.;Lee, C.S.
    • Journal of Animal Science and Technology
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    • v.46 no.1
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    • pp.7-14
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    • 2004
  • The Stearoyl-CoA Desaturase(SCD) is a key enzyme, which converting palmitic acid(16:0) and stearic acid (18:0) to pahnitoleic acid(16:1) and oleic acid(l8:1), respectively. The concentration of oleic acid(18:1) in meat of beef cattle could influence both palatability and perception of meat. This experiment has conducted to determine relationship between the compositions of monounsaturated fatty acids and the SCD mRNA level in bovine liver and loin muscle tissue. The compositions of palmitoleic acid(16:1) and oleic acid(18:1) in loin muscle were 5% and 46% of total lipid and in liver were 2% and 20% of total lipid, respectively. On the other hand, the compositions of palmitic acid(16:0) and stearic acid(18:0) in loin muscle were 25% and 45% of total lipid and in liver were 14% and 43% of total lipid, respectively. The ratio of monounsaturated to saturated fatty acids(the desaturation index) was used as a measure of SCD activity in tissues. The average desaturation index in loin muscle was higher about 3.6-fold than that in liver. The desaturation index of oleate/stearate and palmitoleatelpalmitate in loin muscle were higher 8-fold and 1.8-fold than those in liver, respectively, showing that the substrate specificity of SCD enzyme was very different between liver and muscle tissues. To determine whether the composition of monounsaturated fatty acids in liver and muscle are dependent on SCD expression, SCD mRNA level was examined by RT-PCR analysis. The SCD mRNA level in loin muscle was higher about 3-fold than that in liver. Thus, the quantitative relationship between the desaturation index of fatty acid and SCD mRNA was observed in liver and muscle. The difference in the compositions of monounsaturated fatty acids between bovine liver and muscle tissues may be due to different level of Stearoyl-CoA Desaturase mRNA.

The Profile of Milling Plants in Korea (우리나라 양곡가공공장의 현황분석)

  • 정창주;금동혁;강화석
    • Journal of Biosystems Engineering
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    • v.3 no.1
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    • pp.47-63
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    • 1978
  • This study was conducted to obtain a basic information necessary to assess present rice milling technology in Korea The profiles for milling plants was analyzed by survey work.For the private custom-work mills, which process about 80 percent of domestic rice consumption ,their actual milling test for the identical samples as used for filed mills was conducted. Two rice varieties Japonica and Tongil-type were associated with the experiments. The results are summarized as follows: 1. Analyses for private custom-work mills showed their general aspects as; about 91 percent of the mills belonged to an individual owner ship ; more than 93 percent of the mills was established earlier than 1950 ; about 80 percent of the mills was powered with electric motor; mills having less than two employees were about 75 percent; about 45 percent of the mills provided for warehouse in storing customers cereal grains. 2. The polishers installed in 1,255mills within the surveyed area (7 counties) have been supplied by 44 different domestic manufacturers ;in but about 60 percent of which was supplied by 6major manufacturers. The polishers could be classified into two categories in terms of principles of their polishing actions ;jet-pearler and friction types. About 51 percent of the mills was equipped with the former which has been recognized as giving greater milling recovery than the friction types. 3. Reason for owners of private mills to supplement new machines was due mainly to pgrading their mills to meet the requirements that established by the Government. However, about 60 percent of the mill owners intended to replace with new pearler by their own needs to meet with new high yielding varieties. 4. Processing systems of each previate rice mills surveyed could be classified into three categories, depending upon whether the systems posessed such components as precleaner and paddy separator or not. Only 36.7 percent of mills was installed with both precleanr ad paddy seperrator, 5.0 percent of mills did have neither percleaner nor paddy seperator, and rest of them equipped only one of the two. Hence,it is needed for about 63% of rice miils to be supplemented with these basic facilities to meet with the requirements for the standaized system. 5. Actual milling capacity measured at each field rice mills was shown a wide variation, having range from about 190 to 1,210 kg/hr. The percentages of mills classified according to daily milling capacity based on this hourly capacity were 24.3% for the capacity less than 3 M/T a day; 20.0% for 3-4 M/T; 15.6% for 4-5 M/T; 6.7% for 5-6 M/T; 22.3% for 6-7 M/T; and 11.0% for more than 7 M/T a day. 6. Actual amount of rice processed was about 310 M/T a year in average. About 42% of total milled rice was processed during October to Decembear, which formed a peak demand period for rice mills. The amount of rice milled during January to May was relatively small, but it had still a large amount compared to that during June to September. 7. Utilization rate of milling facility, i. e., percentage of the actual amount of milled rice to the capacity of rice mills, was about 18% on the year round average, about 41% in the peak demand season, and about 10% during June to September. Average number of operating days for mills surveyed was about 250 days a year, and about 21 days a month. 8. Moisture contents of paddy at the time of field mill tests were ranged 14.5% to 19.5% for both Japonica and Tong-i] varieties, majority of paddy grains having moisture level much higher than 1530. To aviod potential reduction of milling recovery while milling and deterioration of milled rice while storage due to these high grain mJisture contents, it may be very important for farmers holding rice to dry by an artificial drying method. 9. Milling recovery of JapJnica varieties in rice mills was 75.0% in average and it was widely ranged from 69.0% to 78.0 % according to mills. Potential increase in milJing recovery of Japonica variety with improvement of mill facilities was estimated to about 1.9%. On the other hand, milling recovery of Tong-il varieties in the field mill tests was 69.8% in average and it ranged from 62% to 77 %, which is much wider than that of Japonica varieties. It is noticed that the average milling recovery of Tong-il variety of 69.8% was much less than that of the Japonica-type. It was estimated th3.t up to about 5.0% of milling recovery for Tong-il variety could be improved by improving the present lo'.ver graded milling technology. 10. Head rice recoveries, as a factor of representing the quality of commercial goods, of Japonica and Tong-il varieties were 65.9% and 53.8% in average, and they were widely ranged from 52% to 73% and from 44% to 65% , respectively. It was assessed that head rice recovery of Japonica varieties can be improved up 3.3% and that of Tong-il varieties by 7.0% by improving mill components and systems.

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The Relationship between Change of Lymphocyte Inositol Monophosphatase mRNA Level by Lithium and Clinical Course in Bipolar Affective Disorder (Lithium에 의한 양극성 기분장애환자의 임파구 Inositol Monophosphatase mRNA 양의 변화와 임상경과)

  • Kim, Seok Hyeon;Lee, Min Soo;Lee, Jang Han
    • Korean Journal of Biological Psychiatry
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    • v.8 no.1
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    • pp.96-105
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    • 2001
  • Objective : Lithium inhibits the action of inositol monophosphatase(IMPase) in phosphoinositide(PI) signal transduction system at therapeutically relevant concentration. The depletion of inositol by lithium itself cannot explain the lithium's therapeutic effect. However, attention has focused on the abnormality of PI signal transduction system as the pathophysiology of bipolar affective disorder(BPD). We investigated whether IMPase mRNA levels of lymphocytes would be different between BPD patients(n=16) and age, sex-matched normal controls(n=16). We also investigated the change of IMPase mRNA level by lithium during 4 weeks to probe the possibility that IMPase mRNA levels could predict the therapeutic response to lithium and clinical course. Method : Relative IMPase mRNA levels in lymphocyte were quantified by reverse transcriptase(RT)-PCR in sixteen drug-free BPD patients and sex, age-matched normal controls. The psychopathology of patients were measured using YMRS (Young Mania Rating Scale) and CGI(Clinical Global Impression). Results : There was no significant difference in IMPase mRNA levels between BPD patients and normal controls. And the IMPase mRNA levels were not significantly changed by 4 week treatment with lithium. However, the basal IMPase mRNA levels were negatively correlated with the changes of CGI after 4 weeks. Furthermore, the patients with relatively high basal IMPase mRNA levels showed much more improvement during 4 weeks. Conclusions : BPD patients and normal controls were not distinguished by lymphocyte IMPase mRNA level. Although we do not support the hypothesis that lymphocyte IMPase activity would be related with the pathogenesis of BPD and the action of lithium, these data raise the possibility that lymphocyte IMPase mRNA levels could function as a predictor of therapeutic response and clinical course of BPD.

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Evaluation of the Effect of Metal Artifacts Varying the Parameters of the Attenuation Map for the artificial Hip Joint in SPECT/CT (SPECT/CT에서 인공고관절에 대한 감쇠보정지도(Attenuation Map)의 매개변수 변화에 따른 금속 인공물(Metal Artifact)의 영향 평가)

  • Kim, Sang Gyu;Kim, Jung Yul;Park, Min Soo;Jo, Seung Hyun;Lim, Han Sang;Kim, Jae Sam
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.2
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    • pp.3-7
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    • 2014
  • Purpose SPECT/CT scan to be performed attenuation correction on the basis of CT induce an overestimation of the site due to the beam hardening artifact by metal cover and reduce the images quality. Therefore, this study using a phantom that has been inserted artificial hip joint investigated that effect on the SPECT/CT image causing by metal artifact for varying the parameters of the Attenuation Map. Materials and Methods Siemens Symbia T16 SPECT/CT equipment was used. Artificial hip joint was inserted to SPECT/PET phantom, 17 mm sphere of Bright Streak area in CT image was filled with Tc-99m so that the radiation activity was 8 times compared to background. And then Hot and Background was measured in varying Wide Beam Coefficient on Attenuation Map and RBR (Region to Background Ratio) of Metal and Non-Metal was calculated and analyzed depending on the presence or absence of the hip joint. Results It tended to hot count of Non-Metal and Metal to increase as the value of the manual mode is increased, hot count ratio with the group of both manual mode 0.5 and 0.4 is the best match. Also, in automatic mode, the ratio of RBRNon-Metal and RBRMetal was 1.135, statistically significant difference was not observed in the manual mode 0.5 and 0.4. Conclusion In the automatic mode of Wide Beam Coefficient in attenuation correction map, it was found that it is over-correction by 13.52%, it was possible to minimize the over-correction by the artifact in 0.5 and 0.4 of manual mode. Further studies should be performed in order to apply to a patient with the help of this and it is considered possible to reduce the over-correction by the metal artifact of an artificial hip joint for Hip-Resurfacing Arthroplasty patients, and to improve the diagnostic performance.

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Reduction of Injection Dose in 18F-FDG Fusion PET (PET-CT 검사에서 18F-FDG 투여량 감소에 대한 고찰)

  • Kim, Jong-Pil;Kim, Jae-Il;Lee, Hong-Jae;Kim, Jin-Eui
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.2
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    • pp.17-21
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    • 2014
  • Purpose With the recent rise of social issue regarding radiation exposure, attention to medical radiation use has been placed under a great spotlight. During PET-CT examination, generally about 40% more of $^{18}F$-FDG is used than EANM recommendation. While maintaining the diagnostic test result, we hope to find optimal injection dose to minimize the $^{18}F$-FDG in patients by utilizing the latest PET-CT scanner which is equiped with the newest technology. Materials and Methods During this experiment, the Biograph Truepoint 40 (siemens, USA) installed in 2007 and mCT 64 (siemens, USA) installed in 2011 were used and evaluated NECR (noise-equivalent counting rate) by using a scatter phantom. For the image quality evaluation of each scanner, we injected 3.7, 4.44 and 5.18 MBq/kg of $^{18}F$-FDG in NEMA IEC Body Phantom and also evaluated SNR between two scanners by using the data acquired at 60, 70, 80, 90, 100, 110 and 120 sec per bed. For the clinical evaluation, actual data of patients who were injected $^{18}F$-FDG 3.7, 4.44, 5.18 MBq/kg were used to compare SNR and draw a final result. Results As a result, mCT 64 peak NECR value was 1.65e+005, which is 10% higher than Turepoint 40. SNR values using the IEC body phantom was 17.9%, 17.4% and 17.1% higher in $^{18}F$-FDG 3.7 MBq/kg, 4.44 MBq/kg and 5.18 MBq/kg. In clinical patients, SNR values of the image mCT 64 was 16.5, which is 25% higher than Turepoint 40 scanner. Conclusion To draw a conclusion from the test result of this experiment, the same quality of SNR could be attained even with 10% reduced injection dose, if when the duration is extended by 10 sec/bed. This optimal result was possible due to enhanced equipment. The NECR (one of the equipment's performance assessment criteria for the scanner) increased by 10% and the SNR (one of the image quality assessment criteria) also increased by 17.5%. Therefore, we can expect to reduce the injection dose without deterioration of image quality. In consequence, it will also help to decrease the patient's anxiety of the radiation exposure.

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Temporal and Spatial Characteristics of Surface Winds over the Adjacent Seas of the Korean Peninsula (한국 주변해역에서의 해상풍의 시공간적 특성)

  • Han, Sang-Kyu;Lee, Heung-Jae;Na, Jung-Yul
    • 한국해양학회지
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    • v.30 no.6
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    • pp.550-564
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    • 1995
  • The temporal and spatial characteristics of wind fields over the neighbouring seas of the Korean peninsula are investigated using 10-years daily wind data during 1978${\sim}$1987 which have been spatially smoothed and low-pass filtered. Long term annual and monthly means are examined for synoptic patterns and spectral analyses are made for temporal variability and spatial coherence. Spatial patterns of the annual mean wind stress and curl have a strong resemblance with those of monthly means during the winter season. Two outstanding periodicities are observed at 1 and 2 cycles per year. The synoptic winds over the study area are highly coherent at both the annual and semi-annual periodicities. However, each basin has its own characteristic spatial pattern. For instance, the prevailing wind during the winter season is northerIy over the northern East Sea (ES), Yellow Sea (YS), and northern East China Sea (ECS), while it is northwesterly over the southern ES and northesterly over the northern ES and southern ECS. At the same time, the wind stress curl is positive over the northern ES and southern ECS, while it is negative over the southern ES, YS and northern ECS. On the other hand, the wind field during the summer season, with its strength being much reduced, is completely different from that during the winter season, and frequent passage of tropical storms provokes large temporal variability over ECS. One remarkable point is that the annual cycle, dominated by the Siberian High, tends to propagate from northeast to southwest, i.e., from northern 25 toward southern ES, YS and ECS, while the semi-annual cycle propagates in the opposite direction, from southwest to northeast. The semi-annual periodicity may reflect development of extratropical cyclones in spring and fall which frequently cross the Korean peninsula. In higher frequencies, there are no dominant periodicities, but local winds over YS and ES are highly correlated for frequencies larger than 0.1 cycles per day and phase difference increases linearly with frequency. This linear increase of phase corresponds to phase speed of 550 and 730 km/d at 0.1 and 0.3 cpd, respectively, The phase speed is apparently coincident with moving speed of extratropical cyclones across the Korean peninsula in the west-east direction.

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Clinical Observations in Vancomycin-Resistant Enterococci Isolated from Pediatric Patients (소아 환아에서 분리된 Vancomycin 내성 장구균에 대한 임상적 고찰)

  • Lee, Dong Woo;Lee, Kyung Jae;Jang, Gwang Choen;Kim, Dong Soo;Lee, Kyung Won;Park, Eun Suk
    • Pediatric Infection and Vaccine
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    • v.8 no.2
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    • pp.199-205
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    • 2001
  • Purpose : Since the first report of vancomycin-resistant enterococci(VRE) in 1986, the resistance to vancomycin in enterococci has been increasingly rapidly. In this study, we investigated the clinical manifestations of pediatric patients with VRE and the pattern of the antibiotic use with increasing the rate of VRE in pediatrics Methods : We studied retrospectively 36 pediatric patients who were isolated VRE from January 1998 to December 2000. We classified patients into ICU and non ICU groups and reviewed species of VRE, specimens in which VRE were first detected and procedures performed before VRE detected. Results : We have found that the number of pediatric patients isolated VRE is increasingly annually in this study. In addition, the number of VRE-isolation in the ICU group and in patients who were operated or who underwent active procedures is much higher than that of in the non ICU group and in patients who were taken medication only. Enterococcus faecium is the main species of VRE. VRE showed high resistance to almost all antibiotics except tetracycline, and resistance was closely related to the duration of hospitalization and history of the antibiotic use. The proportion of the cephalosporin use was higher than any other antibiotic before VRE detection. In contrast, that of teicoplanin was higher than any other antibiotic after VRE detection(P<0.05). The cases of superinfection is higher in the ICU group than in non ICU group. Conclusion : In the hospital level, prevention of nosocomial infection through proper administrative policies, through surveillance of high risk VRE regions and prudent antibiotic use can prevent VRE outbreaks and corresponding side effects.

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