• Journal of Plant Biotechnology
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• v.43 no.1
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• pp.91-98
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• 2016

The growth area of living modified (LM) cotton has steadily increased every year, since its first commercialization in 1996. Development of environmental risk assessment tools and techniques for LM cotton is required for ecosystem safety. We therefore developed multiplex PCR assays for simultaneous detection of two (MON15985, MON531) and four (GHB614, LLCOTTON25, MON88913 and MON1445) LM cotton events approved in Korea, with event specific primer pairs. The PCR reactions were optimized by using event specific primers of six LM cottons at various concentrations. The reactions allows amplification of estimated amplicons of MON15985 (214 bp), MON531 (270 bp), GHB614 (119 bp), LLCOTTON25 (164 bp), MON88913 (276 bp), and MON1445 (389 bp) from multiplex PCR reactions. The multiplex PCR assay developed allowed that two annealing steps (15 cycles at $55^{\circ}C$ and 25 cycles at $60^{\circ}C$) were performed for amplification of distinguished two LM cottons, and only one annealing step (50 cycles at $60^{\circ}C$) was necessary for tetraplex PCR. Primer extension step of all PCR reactions was skipped for time-effective amplification. Our methods suggest that two multiplex PCR assays can be cost-effective and a rapid diagnostic tool for environmental LMO monitoring of six LM cottons.

• Korean Journal of Veterinary Research
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• v.44 no.2
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• pp.241-249
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• 2004

As a part of epidemiologic investigation of tsutsugamushi disease, the wild rodents which were captured in Gyeongnam area were diagnosed with indirect immunofluorescent antibody assay (IFA) and Polymerase Chain Reaction (PCR) to find if they have an antibody against Orientia tsutsugamushi. The conclusion was drawn as followings. (1) The captured 58 wild rodents showed that the subspecies distribution of Apodemus agrarius was 86.2%, Microtus fortis was 8.6% and Crocidura lasiura was 5.2%. (2) The antibody positive rate of O. tsutsugamushi Gilliam, Karp, Karto and Boryong by IFA method was 32.0% in Apodemus agrarius among 50 wild rodents and 40.0% in Microtus fortis among 5 wild rodents, respectively. It was negative in the case of all the 3 Crocidura lasiura. (3) The antibody titers on Apodemus agrarius, Microtus fortis and Crocidura lasiura against Gilliam, Karp, Karto and Boryong were measured between 1:20 and 1:640. The antibody titer against each antigen was in the order Boryong>Gilliam>Karp. (4) O. tsutsugamushi was detected from the blood, spleen and kidney from the artificially infected mice by IFA and PCR. IFA showed the positive response between 3 and 18 days after inoculation. On the other hand, positive response was found from all the samples by PCR. (5) From PCR of the genomic DNA extracted from the blood, spleen and kidney samples of the captured wild rodents, Boryong-specific amplification product with size of 210 bp, which is particular in Boryong, was detected from spleen and kidney samples, but not detected in the blood. (6) Boryong-specific amplification product was detected from spleen and kidney samples which were obtained at 3, 6, 12, 18 and 24 days after the infection with Boryong. But, it wasn't detected from the uninfected samples. (7) From PCR of spleen and kidney samples of the captured wild rodents, it was found that positive rate of O. tsutsugamushi in Apodemus agrarius and Microtus fortis were 25.0% (4/16) and 20.0% (1/5), respectively. From the above results, it can be concluded that Apodemus agrarius resided in Gyeongnam area carried O. tsutsugamushi and PCR method might be a simple, precise, rapid and useful diagnostic tool than IFA for the diagnosis of O. tsutsugamushi.

• The Korean Journal of Nuclear Medicine
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• v.26 no.2
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• pp.290-298
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• 1992

Although early diagnosis of urinary tract infection is important, the radiologic evaluation is still controversial because of the low sensitivity and the lack of cost-effectiveness. This study was carried out to evaluate the clinical utility of high resolution triple head $^{99m}Tc-DMSA$ SPECT imaging in urinary tract infection. We prospectively performed $^{99m}Tc-DMSA$ planar and SPECT imaging, ultrasound of kidney (US), intravenous pyelography (IVP) and voiding cystourethrography (VCU) in all 60 adult patients with UTI [26 with first episode of acute pyelonephritis (APN), 22 with recurrent APN, and 12 persistent asymptomatic pyuria] and 25 normal persons. To assess reversibility of the renal cortical defect (RCD), $^{99m}Tc-DMSA$ SPECT was repeated 1 to 8 months later in those patients with abnormal initial findings. Overall detection rate of $^{99m}Tc-DMSA$ SPECT imaging was 83% (50/60), but planar, US, IVP and VCU showed abnormal findings in 68%, 28%, 32% and 13%, respectively. 25 out of 27 patients with normal or single RCD were all normal in other radioligic studies. Only two patients showed vesicoureteral reflux (VUR) on VCU (grade I) and mild hydronephrosis on IVP. But, high proportion of those with multiple RCD showed abnormal findings on US (17/33), IVP (18/33), and VCU (7/33): 67% in any of these 3 studies. Especially, 3 out 7 patients with VUR showed multiple RCD on $^{99m}Tc-DMSA$ SPECT without any abnormality on IVP or US. 25 normal persons showed normal findings in all studies except one false positive finding on $^{99m}Tc-DMSA$ SPECT imaging. Follow-up $^{99m}Tc-DMSA$ SPECT was done in 28 patients (13 with single RCD, 15 with multiple RCD). All 13 patients with single RCD showed improvement. Those with multiple RCD presented improvement in 4, no change in 10, and aggravation in 1 on follow-up studies. With these results, we conclude: 1) $^{99m}Tc-DMSA$ SPECT imaging is superior to planar imaging, US, IVP or VCU in detection of renal lesion in urinary tract infection. $^{99m}Tc-DMSA$ SPECT is useful as a initial diagnostic tool in adult patients with urinary tract infection. 2) The multiple RCD on $^{99m}Tc-DMSA$ SPECT represent the high probability of irreversible tissue change and need of extensive urological work-up.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2003.07b
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• pp.37-54
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• 2003

It has been recognized that the hen. like its mammalian counterparts. provides young chicks with antibodies as protection against hostile invaders. This system facilitates the transfer of specific antibodies from serum to egg yolk. and provides a supply of antibodies called immunoglobulin Y(IgY) to the developing embryo and the hatched chick. The protection against pathogens that the relatively immuno-incompetent newly hatched chick has. is through transmission of antibodies from the mother via the egg. Egg yolk. therefore. can be loaded with a large amount of IgY against pathogens which can immobilize the existing or invading pathogens during the embryo development or in day-old chicks. Thus. the immunization of laying hens to various pathogens results in production of different antigen-specific IgY in eggs. Egg yolk contains 8~20 mg of immunoglobulins (IgY) per $m\ell$ or 136~340 mg per yolk suggesting that more than 30 g of IgY can be obtained from one immunized hen in a year. By immunizing laying hens with antigens and collecting IgY from egg yolk. low cost antibodies at less than ＄10 per g compared to more than ＄20.000 per g of mammalian IgG can be obtained. This IgY technology opens new potential market applications in medicine. public health veterinary medicine and food safety. A broader use of IgY technology could be applied as biological or diagnostic tool. nut-raceutical or functional food development. oral-supplementation for prophylaxis. and as pathogen-specific antimicrobial agents for infectious disease control. This paper has emphasized that when IgY-loaded chicken eggs are produced and consumed. the specific antibody binds. immobilizes and consequently reduces or inhibits the growth or colony forming abilities of microbial pathogens. This concept could serve as an alternative agent to replace the use of antibiotics. since today. more and more antibiotics are less effective in the treatment of infections. due to the emergence of drug-resistant bacteria.

• Journal of Veterinary Clinics
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• v.29 no.5
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• pp.377-383
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• 2012

Bovine diarrhea is a major economical burden to the bovine industry in Korea. Since multiple infectious agents can be involved in bovine diarrhea, differential diagnosis is essential for effective treatment. Therefore, a panel of two multiplex real-time PCR assays which can simultaneously detect six major bovine enteric pathogens [i.e., bovine viral diarrhea virus (BVDV), bovine coronavirus (BCoV), group A bovine rotavirus (BRV), Salmonella spp., Escherichia coli (E. coli) $K99^+$, and Cryptosporidium parvum] was developed and applied to test 97 fecal samples collected from cattle farms in Korea. In addition, microscopic examination was also preformed on the samples to detect Coccidium oocyst. The estimated sensitivity of the multiplex PCR was 0.1 $TCID_{50}$ for BVDV, BCoV and group A BRV, 5 and 0.5 CFU for E. coli $K99^+$ and Salmonella, respectively, and 50 oocysts for Cryptosporidium. The amplification efficiency of the multiplex PCR ranged between 0.97 and 0.99 for each pathogen. Among 97 samples, 36 samples were positive for at least one of the 6 major pathogens and 6 samples were simultaneously positive for 2 pathogens by the multiplex PCR assay. Coccidium oocysts were also detected in 48 samples, which were all collected from over 1 month old calves. In conclusion, the multiplex real-time PCR panel can be a useful tool for fast and accurate diagnosis of calf diarrhea associated with BVDV, BCoV, group A BRV, E. coli $K99^+$, Salmonella, and/or Cryptosporidium and Coccidium may be an important target which needs to be included in the multiplex PCR panel in the future.

• Weed & Turfgrass Science
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• v.4 no.4
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• pp.376-382
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• 2015

Dollar spot, caused by Sclerotinia homoeocarpa, is the major disease in cool-season turfgrasses. Understanding the distribution of this pathogen in soil and thatch is important to developing disease control strategies. In this study, toothpicks were used to detect S. homoeocarpa in the turfgrass canopy, thatch, and soil at different distances from dollar spot infection centers. The effect of penetrant and contact fungicide applications with different water volumes on distribution of S. homoeocarpa was also investigated. S. homoeocarpa was detected in 100% of samples taken from the leaf canopy, 83.3% in thatch area, and 0% in the soil from within the infection center. S. homoeocarpa was isolated in 100% of samples taken from the edge of the infection center, but was only detected in 13% of the samples taken at 1.5 cm away from the infection center edge. S. homoeocarpa was isolated at a higher frequency in the propiconazole treated plots than those treated with chlorothalonil and was not detected in leaf canopy samples when either fungicides was applied with 6.78 L of water. In conclusion, the toothpick-aided detection technique has improved our understanding of S. homoeocarpa epidemiology and could be used as a diagnostic tool to detect for fungicide resistance on golf courses.

• Tuberculosis and Respiratory Diseases
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• v.44 no.2
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• pp.251-263
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• 1997

Background : The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture. Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in $\beta$subunit of RNA polymerase is main mechanism of resistance. Method : In this study, rpoB gene for the $\beta$subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. Results : All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S531L) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not confirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513~515 were new identified mutations for the first time. Conclusion : Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.

• Investigative Magnetic Resonance Imaging
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• v.13 no.1
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• pp.40-46
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• 2009

Purpose : The purpose of this study was to investigate the quantitative evaluation of the corticospinal tract (CST) at the multiple levels by using functional MRI (fMRI) co-registered to diffusion tensor tractography (DTT). Materials and Methods : Ten normal subjects without any history of neurological disorder participated in this study. fMRI was performed at 1.5 T MR scanner using hand grasp-release movement paradigm. DTT was performed by using DtiStudio on the basis of fiber assignment continuous tracking algorithm (FACT). The seed region of interest (ROI) was drawn in the area of maximum fMRI activation during the motor task of hand grasp-release movement on a 2-D fractional anisotropy (FA) color map, and the target ROI was drawn in the cortiocospinal portion of anterior lower pons. We have drawn five ROIs for the measurement of FA and apparent diffusion coefficient (ADC) along the corona radiata (CR) down to the medulla. Results : The contralateral primary sensorimotor cortex (SM1) was mainly found to be activated in all subjects. DTT showed that tracts originated from SM1 and ran to the medulla along the known pathway of the CST. In all subjects, FA values of the CST were higher at the level of the midbrain and posterior limb of internal capsule (PLIC) than the level of others. Conclusion : Our study showed that co-registered fMRI and DTT has elucidated the state of CST on 3-D and analyzed the quantitative values of FA and ADC at the multiple levels. We conclude that co-registered fMRI and DTT may be applied as a useful tool for clarifying and investigating the state of CST in the patients with brain injury.

• Tuberculosis and Respiratory Diseases
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• v.38 no.2
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• pp.155-163
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• 1991

Bronchoalveolar lavage had been done as the treatment of some diseases such as alveolar proteinsois, bronchiectasis, and severe asthma to remove excessive secretion or mucus. But in the recent decade it has been done as a diagnostic method and a tool to understand and evaluate the pathophysiology of diffuse interstitial lung diseases such as sarcoidosis, pneumoconiosis and hypersensitivity pneumonitis. To analyse the bronchoalveolar fluid, it might be useful to have a standard reference (especially cell counts and differetial count of the cells from bronchoalveolar lavage fluid) of normal person. But it is difficult to study the normal volunteers. We investgated the bronchoalveolar lavage fluid of 48 patients (28 nonsmokers, 20 smokers) who visited Severance Hospital because of minor pulmonary symptoms such as cough and sputum. They did neither complain of dyspnea nor cyanosis, and had normal or unilateral minor lesion on physical examination and chest X-ray. We analysed the recovery rate, viability, total cell count and differential count of the cells in fluid obtained by bronchoalveolar lavage. The following results were obtained: 1) Age ranged from 17 to 72 years-old with the mean age of 36.7; there was no difference of age between the nonsmoker and the smoker gorup. Male to female ratio was 2.43:1 for total group, 1.15:1 for nonsmokers, and 19:1 for smokers. 2) The diagnoses of the patients were undetermined in 41.9%, healed pulmonary tuberculosis in 37.5%, laryngitis or pharyngitis in 10.4% and others in 10.4%. 3) Total cell number of the recovered fluid by bronchoalveolar lavage was significantly higher in male[$9.6{\pm}6.2({\times}10^6)$] than in female[$5.1{\pm}3.0({\times}10^6)$](p<0.05), and there was no significant difference in the total cell number between the smokers and nonsmokers [$9.3{\pm}5.8({\times}10^6)$ vs $7.5{\pm}5.8({\times}10^6)$]. 4) The differential count of the cells from bronchoalveolar lavage fluid had no difference between the nonsmokers and the smokers. 5) There was no correlation between the total cell count and smoking or age. 6) In the smoker group, there was no correlation between the amount of smoking and the total cell count of the bronchoalveolar fluid. In conclusion, it should be careful to regard the patients with symptoms or minor radiologic abnormalities as a control group in bronchoalveolar lavage study and further study of cell analysis in bronchoalveolar lavage will be needed between smoker and nonsmoker in the male and female healthy people.

• Journal of Trauma and Injury
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• v.21 no.1
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• pp.28-35
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• 2008

Purpose: This study was conducted to examine the clinical significance IV-contrasted helical abdomen computed tomography (CT) as a diagnostic screening tool to evaluate hollow viscus injury in blunt abdominal trauma patients. Methods: This is a retrospective study encompassing 108 patients, presenting to Korea University Medical Center (KUMC) Emergency Department (ED) from January 2007 to December 2007, with an initial CT finding suggestive of intra-abdominal injury. An initial non-enhanced abdomen CT was taken, followed by an enhanced CT with intravenous contrast. Patients' demographic data, as well as the mechanisms of injury, were inquired upon and obtained, initial diagnosis, as dictated by specialized radiologists, were added to post-operational (post-OP) findings and to additional CT findings acquired during their hospital stays, and all were combined to arrive at final diagnosis. Initial CT findings were further compared with the final diagnosis, yielding values for sensitivity, specificity, and accuracy, as well as positive and negative predictive values. Patients were further divided into two groups, namely, those that underwent operational intervention and those that did not. The initial CT findings of each group were subsequently compared and analyzed. Results: Initial CT scans revealed abnormal findings in a total of 212 cases - solid organ injuries being the most common finding, as was observed in 97 cases. Free fluid accumulation was evident in another 69 cases. Based on the CT findings, 77 cases (71.3%) were initially diagnosed as having a solid organ injury, 20 cases (18.5%) as having a combined (solid organ + hollow viscus) injury, and 11 cases (10.2%), as having an isolated hollow viscus injury. The final diagnosis however, were somewhat different, with only 67 cases (62.0%) attributed to solid organ injury, 31 cases (28.7%) to combined injury (solid + hollow), and 10 cases (9.3%) to hollow viscus injury. The sensitivity (CI 95%) of the initial helical CT in diagnosing hollow viscus injury was 75.6%, and its specificity was 100%. The accuracy in diagnosing hollow viscus injury was also meaningfully lower compared to that in diagnosis of solid organ injury. Among patients initially diagnosed with solid organ injuries, 10 patients (2 from follow-up CT and 8 from post-OP finding) turned out to have combined injuries. A total of 38 patients underwent an operation, and the proportion of initial CT findings suggesting free air, mesenteric hematoma or bowel wall thickening turned out to be significantly higher in the operation group. Conclusion: Abdominal CT was a meaningful screening test for hollow viscus injury, but the sensitivity of abdominal CT was significantly lower in detecting hollow viscus injury as compared to solid organ injury. This calls for special consideration and careful observation by the ED physicians when dealing with cases of blunt abdominal trauma.