• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1238-1243
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• 2009

The susceptibilities of major enterohemorrhagic Escherichia coli (EHEC) strains to antimicrobial agents and the cytotoxicity of these agents were examined using a total of 38 strains of E. coli O26, O111, and O157, which are the major serogroups of EHEC. Among the 38 strains, 35, 36, and 36 were susceptible to amikacin, imipenem, and norfloxacin, respectively. These antimicrobial agents were further examined to determine their cytotoxicity on Vero cells as well as their effect on the release of Shiga toxins along with trimethoprim/sulfamethoxazole. Each of the E. coli O26, O111, and O157 strains containing both the stx1 and stx2 genes were grown in the absence or presence of these agents at 1/4 minimal inhibitory concentration for 6 h, 12 h, and 18 h. At the concentrations used in this study, none of the agents significantly altered cell count compared with the control group. The level of cytotoxicity in the imipenem group was lower at 12 hand 18 h than their respective controls. In contrast, the level of cytotoxicity in cultures treated with trimethoprim/sulfamethoxazole, norfloxacin, and amikacin was increased. The strains were also examined for the release of Shiga toxins 1 and 2 following treatment with the agents, which were measured by the reversed passive latex agglutination (RPLA) method. The RPLA assay showed a suppression of release of Shiga toxin 2 in the strain cultures containing imipenem. These results indicate that imipenem may be a safe and effective agent for inhibition of these bacteria, which has clinical implications for the treatment of EHEC infections.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.862-868
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• 2014

Certain Escherichia coli (E. coli) strains have the ability to cause diarrheal disease. Five types of diarrheagenic E. coli have been identified, including EHEC, ETEC, EPEC, EAEC, and EIEC. To detect these five diarrheagenic types rapidly, we developed a one-step multiplex PCR (MP-PCR) assay using nine primer pairs to amplify nine virulence genes specific to the different virotypes, with each group being represented (i.e., stx1 and stx2 for EHEC, lt, sth, and stp for ETEC, eaeA and bfpA for EPEC, aggR for EAEC, and ipaH for EIEC). The PCR primers were constructed using MultAlin. The sensitivity and specificity of the constructed multiplex PCR primers were measured using DNA isolated from diarrheagenic E. coli strains representing each group. The limits of detection were as follows: $5{\times}10^1CFU/ml$ for EHEC, $5{\times}10^3CFU/ml$ for ETEC expressing lt and sth, $5{\times}10^4CFU/ml$ for ETEC expressing stp, $5{\times}10^2CFU/ml$ for EPEC, $5{\times}10^4CFU/ml$ for EAEC, and $5{\times}10^2CFU/ml$ for EIEC. To confirm the specificity, C. jejuni, C. perfringens, S. Typhimurium, V. parahaemolyticus, L. monocytogenes, Y. enterocolitica, B. cereus, and S. aureus were used as negative controls, and no amplification was obtained for these. Moreover, this kit was validated using 100 fecal samples from patients with diarrhea and 150 diarrheagenic E. coli strains isolated in Korea. In conclusion, the multiplex PCR assay developed in this study is very useful for the rapid and specific detection of five diarrheagenic E. coli types. This single-step assay will be useful as a rapid and economical method, as it reduces the cost and time required for the identification of diarrheagenic E. coli.

• KSBB Journal
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• v.16 no.6
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• pp.579-591
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• 2001

The cell growth inhibitor effect of cervical cancer cells was investigated by liposome mediated transfection (pRcCMVp53/lipofectin) and by transfection using adenovirus (AdCMVp57). The papilloma virus cancer cell lines we used in this study were HPV16 positive, having inhibiter gene, wild p53 gene, CaSki, SiHa, HPV18 positive HeLa, HeLaS3 and HPV negative C33A, HT3. LacZ gene of E.coli was used as the marker gene for the transfection efficiency. The effect on the inhibition of tumor cell growth was measured by cell count and cell viability though ELISA analysis and MTT assay. The inhibition of tumor cell growth was confirmed by measuring each assay for six days, comparing with the normal control cell growth. The cell growth of cervical cancer calls by transfection was significantly reduced and showed tittle differences among the cell lines. To eliminate the potential problem of Ad(adenovirus) contamination during rAAV production, rAAV can be produced by a triple transfection of vector plasmic, packaging plasmid, and adenovirus helper plasmid. To examine the helper functions of Ad plasmids on the production of rAAV vector, we carried out cotransfection of three plasmids, AAV vector, packaging construct, and Ad helper plasmids. The optimized transfection condition for calcium phosphate method is 25ug of total DNA per 10-cm-diameter plate of 293 cell. We found that rAAV yields peaked at 48hr after Ad infection. The titer of rAAV was measured by the dot blot analysis to measure the number of particles/ml based on the quantification of viral DNA. Recent1y, Kombucha(fungi) was identified as a very potent antileukefic agent. In the present study, effect of natural toxin(plankton) and Kombucha is PSP(GTXI-3, neoSTX), on various MTT assay cervical cancer cell line. Toxin(GTX 1-3, neoSTX) also inhibited the proliferation in primary cervical cancer calls in a dose-dependent toxin concentration. These results showed that toxin was very potent in inhibiting the proliferation of cervical cancer calls in vitro. Toxins and Kombuoha exhibited a dose dependent inhibition of cellular proliferation in cancer cell line.

• Journal of the Society of Naval Architects of Korea
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• v.52 no.1
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• pp.8-18
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• 2015

Reduction of the fuel oil consumption and corresponding greenhouse gas exhausted from ships is an important issue for today's ship design and shipping. Several concepts and devices on the superstructure of a container ship were suggested and tested in the wind tunnel to estimate the air drag reduction. As a preliminary performance evaluation, air drag contributions of each part of the superstructure and containers were estimated based on RANS simulation respectively. Air drag reduction efficiency of shape modification and add-on devices on the superstructure and containers was also estimated. Gap-protectors between containers and a visor in front of upper deck were found to be most effective for drag reduction. Wind tunnel tests had been carried out to confirm the drag reduction performance between the baseline(without any modification) configuration and two modified superstructure configurations which were designed and chosen based on the computation results. The test results with the modified configurations show considerable aerodynamic drag reduction, especially the gap-protectors between containers show the largest reduction for the wide range of heading angles. RANS computations for three configurations were performed and compared with the wind tunnel tests. Computation result shows the similar drag reduction trend with experiment for small heading angles. However, the computation result becomes less accurate as heading angle is increasing where the massively separated flow is spread over the leeward side.

• Journal of Food Hygiene and Safety
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• v.13 no.2
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• pp.143-148
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• 1998

The purpose of this study was to determine the kinetics of paralytic shellfish poison (PSP) destruction at various temperature. The toxic digestive gland homogenate of blue mussel (Mytilus edulis), PSP crude toxin, gonyautoxin group and saxitoxin group were heated at temperature ranging from 90 to $120^{\circ}C$, and then the toxicities were measured in samples heated for various time intervals. The rate constant (k) of the toxic digestive gland homogenate, PSP crude toxin, gonyautoxin group and saxitoxin group were $3.28{\times}10^{-2},\;1.20{\times}10^{-2},\;5.88{\times}10^{-2}\;and\;2.58{\times}10^{-2}\;at\;120^{\circ}C$, respectively. The decimal reduction time (D-value) of the toxic digestive gland homogenate, PSP crude toxin, gonyautoxin group and saxitoxin group were 70, 192, 39 and 89 at $120^{\circ}C$, respectively. These results indicate that PSP crude toxin is most heat-stable of 4 types of PSP toxins and PSP toxin are more heat-stable than food poisoning bacteria and spores. The retorting condition to reduce PSP toxicity below quarantine limit ($80\;\mu\textrm{g}/100\;g$ in Korea and America, 4 MU/g in Japan) could be calculated by rate constant. For example, the digestive gland homogenate having a initial toxicity of $200\;\mu\textrm{g}/100\;g$ could have toxicity below quarantine limit when heated at $90^{\circ}C$ for 129 min., $100^{\circ}C$ for 82 min., $110^{\circ}C$ for 48 min. and $120^{\circ}C$ for 28 min. These results suggest that commercial retorting condition ($115^{\circ}C$ for 70 min) in Korea is enough to reduce toxicity below quarantine limit from initial toxicity of $200\;\mu\textrm{g}/100\;g$. From these results, the quarantine limit of PSP-infested shellfish for canning can be level up to raw score of $200\;\mu\textrm{g}/100\;g$.

• Journal of Dairy Science and Biotechnology
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• v.33 no.1
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• pp.59-67
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• 2015

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is well-characterized as an important food-borne pathogen worldwide and causes human diseases such as diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome (HUS) by producing shiga-like toxin (Stx). It has been reported that a number of dairy foods, including cheese, can act as the source of EHEC O157:H7 infections. In addition to the toxicity of Stx, recently it has been indicated that EHEC O157:H7 possesses virulence factors related to attachment, quorum sensing, and biofilms. Moreover, these novel virulence factors might become critical points to be considered in the future production of food derived from animals. Here, we review the evidences that support these insights on new virulence factors and discuss the potential mechanisms mediating the pathogenesis of EHEC O157:H7 in the dairy food industry.

• Korean Journal of Ecology and Environment
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• v.50 no.1
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• pp.137-147
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• 2017

The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.

• The Journal of Korean Society for Radiation Therapy
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• v.27 no.1
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• pp.31-43
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• 2015

Purpose : Stereotactic body radiation therapy (SBRT) has proved its efficacy in several patient populations with primary and metastatic limited tumors. Because SBRT prescription is high dose level than Conventional radiation therapy. SBRT plan is necessary for effective Organ at risk (OAR) protection and sufficient Planning target volume (PTV) dose coverage. In particular, multi-target cases may result excessive doses to OAR and hot spot due to dose overlap. This study evaluate usefulness of Volumetric modulated arc therapy (VMAT) in dosimetric and technical considerations using Flattening filter free (FFF) beam. Materials and Methods : The treatment plans for five patients, being treated on TrueBeam STx(Varian$^{TM}$, USA) with VMAT using 10MV FFF beam and Standard conformal radiotherapy (CRT) using 15MV Flattening filter (FF) beam. PTV, liver, duodenum, bowel, spinal cord, esophagus, stomach dose were evaluated using the dose volume histogram(DVH). Conformity index(CI), homogeneity index(HI), Paddick's index(PCI) for the PTV was assessed. Total Monitor unit (MU) and beam on time was assessed. Results : Average value of CI, HI and PCI for PTV was $1.381{\pm}0.028$, $1.096{\pm}0.016$, $0.944{\pm}0.473$ in VMAT and $1.381{\pm}0.042$, $1.136{\pm}0.042$, $1.534{\pm}0.465$ in CRT respectively. OAR dose in CRT plans evaluated 1.8 times higher than VMAT. Total MU in VMAT evaluated 1.3 times increase than CRT. Average beam on time was 6.8 minute in VMAT and 21.3 minute in CRT. Conclusion : VMAT for SBRT in multi-target liver cancer using FFF beam is effective treatment techniqe in dosimetric and technical considerations. VMAT decrease intra-fraction error due to treatment time shortening using high dose rate of FFF beam.

• Journal of the Korean Society of Radiology
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• v.17 no.3
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• pp.481-487
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• 2023

In this study, we analyzed the incidence of side effects of photoneutron dose according to the number of beams during intensity-modulated radiotherapy of prostate cancer using 15 MV. The radiation treatment plan design for intensity-modulated radiation therapy for prostate cancer was established with a prescription dose of 220 cGy per dose and a total of 7260 cGy for 33 treatments. The linear accelerator used in the experiment is Varian's True Beam STx (Varian, USA). Photoneutron dose was generated by using 15 MV energy in the planning target volume (PTV). The treatment plan was designed with IMRT 5, 7, and 9 portals using the Eclipse System (Varian Ver 10.0, USA). An optically stimulated luminescence albedo neutron dosimeter (Landauer Inc., USA) was used to measure photoneutron dose. IMRT 5 portals, 1.7 per 1,000, 7 portals, 1.8 per 1,000, 9 portals, 2.0 per 1,000 were calculated as the probability of experiencing side effects on the thyroid gland due to photoneutron dose. This study studies the risk of secondary radiation exposure dose that can occur during intensity-modulated radiation therapy, and it is considered that it will be used as useful data in relation to stochastic effects in the future.

• Journal of Welding and Joining
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• v.27 no.3
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• pp.85-91
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• 2009

A roll projection welding machine is introduced to fabricate metallic sandwich plate consisting of a structured inner sheet with projection-like shape and a pair of skin sheets. To fabricate the metallic sandwich plate of consistent and good quality, two process monitoring methods are introduced; dynamic resistance monitoring and skin sheet temperature monitoring. Dynamic resistance monitoring has no time delay but gives only averaged value over plate width. Skin sheet temperature monitoring has certain amount of time delay but is good for predicting weld quality of specified position. By the two complementary monitoring methods, the characteristics of the new welding process is successfully understood.