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Cloning of a Glutathione S-Transferase Decreasing During Differentiation of HL60 Cell Line (HL6O 세포주의 분화 시 감소 특성을 보이는 Glutathione S-Transferase의 클로닝)

  • Kim Jae Chul;Park In Kyu;Lee Kyu Bo;Sohn Sang Kyun;Kim Moo Kyu;Kim Jung Chul
    • Radiation Oncology Journal
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    • v.17 no.2
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    • pp.151-157
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    • 1999
  • Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

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Effects of Heat-stress on Rumen Bacterial Diversity and Composition of Holstein Cows (고온 스트레스 영향에 따른 홀스타인종 젖소의 반추위내 미생물 균총 변화)

  • Kim, Dong Hyeon;Kim, Myung Hoo;Kim, Sang Bum;Ha, Seung Min;Son, Jun Kyu;Lee, Ji Hwan;Hur, Tai Young;Lee, Jae Yeong;Park, Ji Hoo;Choi, Hee Chul;Lee, Hyun Jeong;Park, Beom Young;Ki, Kwang Seok;Kim, Eun Tae
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.39 no.4
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    • pp.227-234
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    • 2019
  • This study was performed to investigate the effect of heat-stressed environment on rumen microbial diversity in Holstein cows. Rectal temperature and respiration rate were measured and rumen fluid was collected under normal environment (NE; Temperature humidity index (THI)=64.6) and heat-stressed environment (HE; THI=87.2) from 10 Holstein cows (60±17.7 months, 717±64.4 kg) fed on the basis of dairy feeding management in National Institute of Animal Science. The rumen bacteria diversity was analyzed by using the Illumina HiSeqTM 4000 platform. The rectal temperature and respiratory rate were increased by 1.5℃ and 53 breaths/min in HE compared to that in NE, respectively. In this study, HE exposure induced significant changes of ruminal microbe. At phylum level, Fibrobacteres were increased in HE. At genus level, Ruminococcaceae bacterium P7 and YAD3003, Butyrivibrio sp. AE2032, Erysipelotrichaceae bacterium NK3D112, Bifidobacterium pseudolongum, Lachnospiraceae bacterium FE2018, XBB2008, and AC2029, Eubacterium celulosolvens, Clostridium hathewayi, and Butyrivibrio hungatei were decreased in HE, while Choristoneura murinana nucleopolyhedrovirus, Calothrix parasitica, Nostoc sp. KVJ20, Anabaena sp. ATCC 33047, Fibrobacter sp. UWB13 and sp. UWB5, Lachnospiraceae bacterium G41, and Xanthomonas arboricola were increased in HE. In conclusion, HE might have an effect to change the rumen microbial community in Holstein cows.

Development of Assay Methods for Enterotoxin of Escherichia coli Employing the Hybridoma Technology (잡종세포종기법을 이용한 대장균의 장독소 측정법 개발)

  • Kim, Moon-Kyo;Cho, Myung-Je;Park, Kyung-Hee;Lee, Woo-Kon;Kim, Yoon-Won;Choi, Myung-Sik;Park, Joong-Soo;Cha, Chang-Yong;Chang, Woo-Hyun;Chung, Hong-Keun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.151-161
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    • 1986
  • In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

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국제 수학 올림피아드 참가 후보자들을 위한 상황대처훈련에 관한연구

  • 김보경;조성희;이군현
    • Proceedings of the Korean Society for the Gifted Conference
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    • 1994.08a
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    • pp.1.2-37
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    • 1994
  • Currently Korea encourages gifted high schoolers and junior high schoolers to participate in international achievement contests such as International Olympiads. Participants for these contests are selected nationwide among gifted students in areas of mathematics, physics, chemistry, and others. They go through a series of screening tests and programs. One of the screening processes IS Korean Olympiad School, which provides study programs each summer for student-candidates prior to following year's International Olympiads. Approximately 40 students of high schools and junior high schools, in each subject of study, gather at Korean Olympiad Summer School, and they go through intensive study programs during a short period of time. Out of 40 candidates,' less than 10 students are finally selected to participate in International Olympiads. In this study, a psycho-educational program called "Situation Coping Training Program" was developed to enhance ahievement motivation for these student-candidates. This study was to see if this training program actually improved their cognitive, emotive motivation factors, and to see how this training program affected their achievement level. Training was administered for five days. This training program was found effective for participants to increase self-efficacy, internal locus of control, and anxiety copmg. These cognitive and emotive motivation factors, other than intelligence, were found to have positive relationship with achievement level, of which self-efficacy and attribution style of students were found as two best predictors of achievement. This training program was perceived as necessary. by participants, and helpful for recovering self-confidence and self-control as well as coping pressure. Suggestions were made that this kind of training program be administered as a regular curriculum in preparative study programs such as Korean Olympiads, since cognitive, emotive motivation factors are related with achievement, and furthermore, be utilized in all gifted education programs in Korea. in Korea.

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The Preference and Inhibitory Effect of Root Vegetables on β-Glucuronidase and Tryptophanase of Human Intestinal Bacteria (근채류의 기호도와 장내세균의 유해효소 억제효과)

  • Han, Myung Joo;Kim, Na Young
    • Korean journal of food and cookery science
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    • v.15 no.6
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    • pp.555-564
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    • 1999
  • The objective of this study was to investigate the preference of root vegetables and the inhibitory effect of the vegetables on harmful enzymes of intestinal bacteria. Two hundred fifty respondents in Seoul area surveyed to obtain information from Sep. 30 to Oct. 30, 1998. Respondents preferred Inpuomoea batatas (sweet potato, 4.05), Solanum tuberosum(potato, 3.97), Allium cepa(onion, 3.68), Codonopsis lanceolata(3.64) and Raponus sativus(redish, 3.60). The growth of B. breve K-110 was effectively increased by adding 0.5% extract of Solanum tuberosum(139%), Codonopsis lamceolate(145%), Dioscorea japonica(164%), Colocisia antiquorum(144%) extract to the medium. B. breve K-100 for beneficial bacteria, and E. coli HGU-3 or Bacteroides JY-6 for harmful bacteria were used to determine the inhibitory effect of root vegetables on harmful intestinal enzymes after co-culturing harmful and beneficial bacteria. The extract of Solanum tuberosum, Codonopsis lanceolata, Dioscorea japonica (yam) and Colocisia antiquorum (taroes) showed inhibitory effect on ${\beta}$-glucuronidase and tryptophanase of intestinal bacteria. The macromolecules were isolated from Solanum tuberosum, Codonopsis lanceolata, Dioscorea japonica and Colocisia antiquorum by Sephadex G-100 column chromatography. By adding these isolated marcromolecules to the medium, the growth of B. breve K-100 were also increased and high inhibitory effects on the ${\beta}$-glucuronidase and tryptophanase were measured. These results suggested that the harmful enzymes of intestinal bacteria were inhibited by consuming Solanum tuberosum, Codonopsis lanceolata, Dioscorea japonica and Colocisia antiquorum. Therefore, they could prevent gastrointestinal diseases.

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Packing effects on the intracavitary radiation therapy of the utaine lervix cancer ($^{192}Ir$source를 이용할 자궁경부암 강내치료시 사용하는 packing의 효과에 대한 고찰)

  • Cho, Jung-Kun;Lee, Du-Hyun;Si, Chang-Kun;Choi, Yoon-Kyung;Kim, Tae-Yoon
    • The Journal of Korean Society for Radiation Therapy
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    • v.16 no.1
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    • pp.73-77
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    • 2004
  • Purpose of the radio-therapy is maximize the radiation dose to the tumor while minimizing the dose to the critical organ. Carcinoma of the uterine cervix treatment are external irradiation or an interstitial brachtheraphy make use of isotope. Brachytherapy is a method of radiotherapy in advantage to achieve better local control with minimum radiation toxicity in comparison with external irradiation because radiation dose is distributed according to the inverse square low of gamma-ray emitted from the implanted sources. Authors make use of the patients data which 192Ir gives medical treatment intrcavity. Intracavitary radiation of the uterine cervix cancer, critical organ take $20\%$ below than exposure dose of A point in the ICRU report. None the less of the advice, Radiation proctitis and radiation cystitis are frequent and problematic early complications in patients treated with radiation for the uterine cervix cancer. In brachytherapy of uterine cervical cancer using a high dose rate remote afterloading system, it is of prime importance to deliver a accurate dose in each fractionated treatment by minimizing the difference between the pre-treatment planned and post-treatment calculated doses. Use of packing to reduce late complications intracavitary radiation of the uterine cervix cancer. Bladder and rectum changes exposure dose rate by radiotherphy make use of packing.

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Intestinal Atresia - The Second National Survey (선천성 장폐쇄증 2차 조사 - 대한소아외과학회 회원을 주 대상으로 한 전국조사 -)

  • Kim, Seong-Chul;Kim, D.Y.;Kim, S.Y.;Kim, I.K.;Kim, I.S.;Kim, J.E.;Kim, J.C.;Kim, H.Y.;Park, K.W.;Park, W.H.;Park, J.Y.;Seo, J.M.;Seol, J.Y.;Oh, S.M.;Oh, J.T.;Lee, N.H.;Lee, M.D.;Lee, S.K.;Lee, S.C.;Chung, S.Y.;Jung, S.E.;Jeong, Y.J.;Jung, P.M.;Choi, K.J.
    • Advances in pediatric surgery
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    • v.16 no.1
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    • pp.1-10
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    • 2010
  • The members of the Korean Association of Pediatric Surgeons conducted a retrospective study of two hundred and twenty-two cases of intestinal atresia for the period from January 1, 2007 to December 31, 2009. Seventeen hospitals were involved. There were 76 duodenal, 65 jejunal, and 81 ileal atresias (3 colonic). The male to female ratio was 0.85:1 in DA and 1.34:1 in JIA. Ninety-four patients(43.3 %) were premature babies (DA 40.3 %, JA 64.6 %, IA 28.8 %), and 70 babies (32.0 %) had low birth weight (DA 38.7 %, JA 44.4 %, IA 16.0 %). Antenatal diagnosis was made in 153 cases (68.9 %). However, 27 infants (17.6 %) with antenatal diagnosis were transferred to the pediatric surgeon's hospitals after delivery. Maternal polyhydramnios was observed in 81 cases (36.59 %) and most frequent with proximal obstruction. In forty-four cases (19.8 %), only simple abdominal film was taken for diagnostic study. The associated malformations were more frequently observed in DA - 61.8 % in DA and 22.6 % in JIA. Meconium peritonitis, small bowel volvulus and intussusception were more frequently associated with ileal atresia. The overall mortality rate was 3.6 %. (Abbreviations: DA;duodenal atersia, JA;jejunal atresia, JIA;jejunoileal atresia, IA;ileal atrsia).

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Food Component Characteristics of Red-tanner Crab (Chionoecetes japonicus) Paste as Food Processing Source (식품가공소재로서 게 페이스트의 식품성분 특성)

  • Kim, Hye-Suk;Park, Chan-Ho;Choi, Seung-Geal;Han, Byung-Wook;Kang, Kyung-Tae;Shim, Nam-Hyuk;Oh, Hyeon-Seok;Kim, Jin-Soo;Heu, Min-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.1077-1081
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    • 2005
  • The objective of this study was to investigate the components and nutritional quality of red-tanner crab (Chionoecetes japonicus) paste in order to explore possibility for food processing source such as surimi gel containing crab paste. Yield of crab paste was $30\%$ from whole body after crushing and dehydrating. Crude protein contents $(9.5\%)$ of crab paste was lower than that $(13.1\%)$ of crab muscle, but fat $(0.5\%)$ and ash contents $(8.0\%)$ of paste were higher than $0.2\%\;and\;1.3\%$ of crab muscle, respectively. Volatile basic nitrogen (VBN) content of the crab paste was lower than those of the edible parts. Total amino acid content (9,497mg/l00g) of paste was lower than that (12,980mg/100g) of muscle. Aspartic acid, glutamic acid, lysine and leucine were the predominant amino acids in the protein fraction. The calcium content (6,539mg/l00g) was higher than those of phosphorus (579mg/100g), and potassium (793mg/100g) while manganese and iron were present in trace amounts. Major fatty acids of total lipid were 16:0, 18:1n-9, 20:5n-3 and 22:6n-3, and no difference of composition between paste and muscle. Sensory evaluation showed that scores of color and flavor of $15\%$ substituted surimi gel increased significantly when compared to surimi gel without crab paste (p<0.05). From the above results, the addition of crab paste enhanced nutrition and functionality of surimi gel.

Sensory Evaluation and Bioavailability of Red Ginseng Extract(Rg1, Rb1) by Complexation with ${\gamma}$-Cyclodextrin (${\gamma}$-cyclodextrin으로 포접한 홍삼추출물의 관능평가 및 Rg1, Rb1의 생체이용율)

  • Lee, Seung-Hyun;Park, Ji-Ho;Cho, Nam-Suk;Yu, Heui-Jong;You, Sung-Kyun;Cho, Cheong-Weon;Kim, Dong-Chool;Kim, Young-Heui;Kim, Ki-Ho
    • Korean Journal of Food Science and Technology
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    • v.41 no.1
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    • pp.106-110
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    • 2009
  • In order to reduce the bitter taste and improve the bioavailability of red ginseng extract(RGE), inclusion complexes (RGE-CD) of the extract with ${\alpha}-,\;{\beta}-,\;{\gamma}$-cyclodextrin were prepared and studied for their sensory quality and bioavailability compared to RGE. By complexation, the bitter taste-reducing efficacies of ${\alpha}$-CD and ${\beta}$-CD were much lower than that of ${\gamma}$-CD. In comparative sensory analysis for the bitter taste, RGE-${\gamma}$-CD10, prepared using 10%(w/w) of ${\gamma}$-CD, showed a score of 1.93(decreased by about 78%) compared to RGE as the control. In addition, in sensory analysis for flavor, RGE-${\gamma}$-CD10showed a score of 5.60. Upon increasing the amount of ${\gamma}$-CD to 15%(w/w) and 20%(w/w), respectively, the bitter taste of RGE-${\gamma}$-CD was removed and the flavor of RGE disappeared(scores of 2.67 and 1.67, respectively). Therefore RGE-${\gamma}$-CD10 was chosen as an optimum. The same dosages of RGE and RGE-${\gamma}$-CD10 were orally administered to SD(Sprague-Dawley) rats on a saponin basis, and the plasma concentrations of ginsenoside Rg1 and Rb1 were measured over time to estimate the average AUC(area under the plasma concentration versus time curve) of the ginsenosides. After the oral administration, there were no significant differences in the AUC values of the RGE and RGE-${\gamma}$-CD 10 groups for ginsenoside Rg1. However, AUC values for ginsenoside Rb1 were $25.8{\mu}g{\cdot}hr/mL$ in the RGE group and $81.5{\mu}g{\cdot}hr/mL$ in the RGE-${\gamma}$-CD 10 group, respectively. Therefore, the bioavailability of ginsenoside Rb1 in the RGE-${\gamma}$-CD 10 group was significantly higher by up to 315% compared with that in the RGE group(p = 0.0029). These results show that the bitter taste of RGE can be simultaneously removed by the complexation of RGE and ${\gamma}$-CD(RGE-${\gamma}$-CD) along with increased bioavailability.

Susceptibility tests of oral antibiotics including cefixime against Escherichia coli, isolated from pediatric patients with community acquired urinary tract infections (소아 원외 요로감염 환아에서 분리된 E. coli에 대한 cefixime을 포함한 경구 항생제의 감수성 연구)

  • Lee, Soo Young;Lee, Jung Hyun;Kim, Jong Hyun;Hur, Jae Kyun;Kim, Sun Mi;Ma, Sang Hyuk;Kang, Jin Han
    • Clinical and Experimental Pediatrics
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    • v.49 no.7
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    • pp.777-783
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    • 2006
  • Purpose : Urinary tract infection(UTI) is one of the most frequent infections in children. E. coli is the most frequent etiological micropathogen in pediatric community UTI, and E. coli has developed resistance to many antibiotics, highlighting the need for regular surveys of this organism resistant patterns in the community. The aim of this study was to determine the oral antibiotic susceptibility patterns of E. coli, isolated from pediatric patients with uncomplicated community acquired UTI. Methods : E. coli isolates, obtained from pediatric patients with uncomplicated community acquired UTI between October in 2004 to September in 2005. And minimal inhibitory concentrations(MICs) of oral aminopenicillins and beta-lactamase inhibnitors(ampicillin, amoxacillin, ampicillin-sulbactam), oral cephalosporins(cefaclor, cefixime) and sulfa drug(trimethoprime-sulfamethoxazole) were performed according to the National Committee for Clinical Laboratory Standards(NCCLS) guide line. Results : Total 211 organisms were isolated from pediatric out-patients with community UTI. E. coli was the most common organism(89 percent), followed by E. fecalis, Proteus species, S. aureus, M. morganii, and P. aeruginosa. The resistant rates of aminopenicillins and beta-lactamase inhibitors, cefaclor and sulfa drug to E. coli were very high. But, the resistant rate of cefixime was markedly low, and ESBL strains were isolated with small rates. Conclusion : Our study results suggest that aminopenicillins, cefaclor and sulfa drug may not be useful as first line empirical antibiotics to treat pediatric patients with community UTI in Korea. But, 3rd generation cephalosporin such as cefixime can be used as effective second line antibiotics after primary treatment failure, also may be useful as an empirical first line antibiotic. Finally, we conclude that a continuous surveillance study to monitor susceptibility patterns of E. coli in community UTI will be needed for the standard guide lines of empirical oral antibiotic treatment.