• Horticultural Science & Technology
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• v.35 no.1
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• pp.121-130
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• 2017

This study was carried out to investigate a method for producing cultured virus - free ovules for breeding high - quality Citrus cultivars. Ovules from the immature fruits of three citrus cultivars native to Jeju (Dongjeongkyool, Cheongkyool, and Jikak) and two cultivars of Citrus unshiu Marc. (Miyagawa wase and Haryejosaeng) that were thought to be infected with Citrus tristeza virus (CTV) were cultured on MS2 medium (Murashige - Skoog [MS] basal medium containing $500mg{\cdot}L^{-1}$ malt extract, $50g{\cdot}L^{-1}$ sucrose, $1.0 mg{\cdot}L^{-1}$ kinetin, and $8g{\cdot}L^{-1}$ agar). After four weeks of culture, 10, 21, 13, 5, and 7 somatic embryos and 2, 4, 2, 4, and 5 white callus cells (surrounding green somatic embryos) were obtained from Dongjeongkyool, Cheongkyool, Jikak, Miyagawa wase, and Haryejosaeng, respectively. After six weeks of culture, somatic embryos were obtained from cultured cells grown on MT basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), lactose ($70g{\cdot}L^{-1}$), and agar ($16g{\cdot}L^{-1}$). Over 60% of the somatic embryos from citrus cultivars native to Jeju developed into normal plants on MS basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), sucrose ($50g{\cdot}L^{-1}$), and agar ($8g{\cdot}L^{-1}$) after 10 weeks of culture. Normal plants were regenerated from two Citrus unshiu Marc. cultivars on MT basal medium supplemented with sorbitol (1.0 M), galactose (1.0 M), $GA_3$ ($1.0mg{\cdot}L^{-1}$), and Gelrite ($3g{\cdot}L^{-1}$). The absence of virus in plants generated from cultured ovules was confirmed by RT - PCR and antigen - antibody reactions. Therefore, virus - free Citrus cells can be obtained for breeding high - quality citrus cultivars using the biotechnological technique evaluated in this study.

• Tuberculosis and Respiratory Diseases
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• v.48 no.3
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• pp.315-323
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• 2000

Background : An immunochromatographic assay (ICT Diagnostics) which facilitates the diagnosis of tuberculosis(TB) by detecting serum antibodies mainly directed against specific 38KDa of Mycobacterium tuberculosis has come into the market. The test consists of a cardboard folding device containing nitrocellulose strip and absorbent pads. The whole procedure is completed within 15 min and does not require any additional equipment. The test has been reported to be sensitive and specific in diagnosing active TB. Thus the test had been evaluated with sera from TB patients and TB-free subjects. Method : Sera from patients with active pulmonary tuberculosis(40 sputum positives for Mycobacterium tuberculosis, 79 sputum negatives, and 3 extrapulmonary tuberculosis) were obtained from the Double-Cross Chest Clinic of the Korean National Tuberculosis Association (KNTA) in Seoul. The control group consisted of TB-free 68 subjects(21 children under 7 years old and 47 healthy staff members of KNTA). Results : Nine out of 68(13.2%) TB-free controls had positive antibody response. Total 106 of 122(86.9%) radiologically active patients had positive antibodies while 16 (13.1%) showed negative reaction. Antibody was detected in 38 of 40(95.0%) sputum positive patients and 68 of 82(82.9%) sputum negative patients who were under the antituberculosis chemotherapy. The sensitivity and specificity were all 87% and the positive predictive value was 92.2% while the negative predictive value was 78.7%, when the prevalence of TB in the sample was 64.2%. Our results clearly show that the detection of antibodies which mainly react with the 38KDa antigen of M. tuberculosis is not suitable as the first-line method of diagnosis but considered only as an adjunctive test to standard techniques of tuberculosis diagnosis. when considering its high false positivity.

• Investigative Magnetic Resonance Imaging
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• v.9 no.2
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• pp.109-116
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• 2005

Purpose : To determine the usefulness of additional Mn-DPDP MRI for preoperative evaluation of the patients with colorectal cancers by comparison of dual-phase CT scan, Mn-DPDP enhanced MRI and combination of CT and MRI. Materials and Methods : Fifty-three colorectal cancer patients with 92 metastatic nodules underwent dualphase (arterial and portal) helical CT scan and Mn-DPDP MRI prior to surgery. The indication of MRI was presence or suspected of having metastatic lesions at CT scan and/or increased serum carcinoembryonic antigen (CEA) levels (10 ng/mL or more). The diagnosis was established by the combination of findings at surgery, intraoperative ultrasonography, and histopathologic examination. Two radiologists interpreted CT, MRI, and combination of CT-MRI at discrete sessions and evaluated each lesion for location, size, and intrinsic characteristics. The lesions were divided into three groups according to their diameter; 1cm<, 1-2 cm, and >2 cm. Diagnostic accuracy was evaluated using the alternative-free response receiver operating characteristic method. Detection and false positive rate were also evaluated. Results : In the lesions smaller than 1 cm, detection rate of combined CT-MRI was superior to CT or MRI alone (82%, p=0.036). The mean accuracy (Az values) of combined CT and MRI was significantly higher than that of CT in the lesions smaller than 2 cm (1 cm<, p=0.034; 1-2 cm, p=0.045). However, there was no significant difference between MRI and combined CT-MRI. The false positive rate of CT was higher than those of combined CT-MR in the lesions smaller than 1 cm (28%, p=0.023). Conclusion : Additional MRI using Mn-DPDP besides routine CT scan was helpful in differentiating the hepatic lesions (<2 cm) and could improve detection of the small hepatic metastases (<1 cm) from colorectal carcinoma.

• Journal of Veterinary Clinics
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• v.29 no.4
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• pp.309-314
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• 2012

The study was undertaken to evaluate sensitivity and specificity of rapid Avian Influenza (AI) and Newcastle Disease virus (NDV) combo antigen kits from field samples of domestic (broiler and layer chicken, native chicken) and semi-domestic (duck, goose, pigeon and quail) birds of Bangladesh. Samples were collected from naturally infected AI suspected domestic and semi-domestic birds of five different outbreak areas in Bangladesh. From each area two birds were selected for sampling, and from each bird three types of samples (tracheal, cloacal and oro-nasal swabs) were collected. A total of 210 field samples from a total of 70 birds were collected and tested using AI and NDV combo antigen rapid diagnostic kits in the study. All three different samples from a bird showed similar pattern of reaction. Out of 210 samples, 15 samples (5 birds), 63 samples (21 birds) and 27 samples (9 birds) were positive for AIV, NDV and both for AIV and NDV, respectively; whereas the remaining birds were negative for either AIV or NDV in this screening test. Among the five AIV positive, a layer chicken from wet market in Mymensingh, Netrokona, Gibandha and Kurigram and a native chicken from wet market in Kurigram area was positive to AIV. The semi-domestic birds are either positive to NDV or free from both AIV and NDV. This study revealed that the AIV and NDV rapid diagnostic kits could be effectively use to diagnose the respective virus in trachea, oro-nasal and cloacal samples simultaneously. AIV-NDV combo Ag test result clearly indicates that the test kit designed for AIV and NDV could diagnose the disease rapidly with less effort and higher scientific know how which could be used for the detection of AIV and NDV using field samples in large scale.

• Radiation Oncology Journal
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• v.29 no.1
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• pp.11-19
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• 2011

Purpose: To evaluate the pathological and clinical effects of preoperative chemoradiation (CCRT) in cases of locally advanced rectal cancer and to determine the predictive factors for tumor downstaging. Materials and Methods: From March 2004 to August 2008, 33 patients with locally advanced rectal cancer were treated with preoperative CCRT. Twenty-eight patients (84.8%) were treated using a concomitant boost technique while five (15.2%) patients were treated using a cone down boost technique. All patients received 50.4 Gy of irradiation and concurrent chemotherapy with 5-fluorouracil. The median follow-up duration was 24.2 months (range, 9.8 to 64.7 months). Results: Thirty-one (93.9%) patients underwent surgery. Twenty-four patients (72.7%) underwent anal sphincter-preserving surgery. The 3-year disease free survival (DFS) and overall survival rates were 63.4% and 78.8%, respectively. Post-operative factors were more important for DFS. Pathologic N stage, margin status, and pathologic differentiation were significant prognostic factors (p=0.001, 0.029, 0.030). Tumor size and lymphovascular invasion were also associated with marginal significance (p=0.081, 0.073). However, only pre-treatment T stage was a significant pre-operative factor (p=0.018). The complete pathological response rate was 9.1 %. T-downstaging was observed in ten (30.3%) patients, whereas N-downstaging was found in 24 (72.7%) patients. Pre-treatment T stage and the interval between CCRT and operation were the predictive factors for downstaging in a univariate analysis (p=0.029, 0.027). Pre-treatment carcinoembryogenic antigen was also associated with marginal significance (p=0.068). Conclusion: The survival of rectal cancer patients can be better determined based on post-operative findings. Therefore, pre-operative CCRT for downstaging of the tumor seems to be important. Pre-treatment T stage and the interval between CCRT and operation can be used to predict downstaging.

• Korean Journal of Veterinary Research
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• v.25 no.1
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• pp.7-17
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• 1985

Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.756-762
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• 1998

The purpose of this study was to investigate the confirmation and prevalence of porcine cytomegalovirus (PCMV) infection of pigs in Korea using enzyme-linked immunosorbent assay (ELISA). Four hundred-eighty one sera tested were collected from the areas of Kyonggi, Kangwon, Chungcheong, Cholla, Gyongsang and Cheju during the year of 1991 to 1997 except 1994. PCMV antigen, OF-1 strain, for ELISA, was prepared 19-PFT-F cell line originated from porcine fallopian tube. Positive control was used by sera made from the specific pathogen free piglets which were infected with OF-1 strain. Three hundred-sixty seven of 481 sera (76.3%) were positive against PCMV. Antibody titers of these seropositives were classified by 129 (26.8%) cases in more than 1 : 12,800, 77 (16.0%) in 1 : 6,400, 76 (15.8%) in 1 : 3,200, 44 (9.2%) in 1 : 1,600, and 41 (8.5%) in 1 : 800, respectively. Also, the seropositive pigs were divided by 87.4% (76/87) in older than 6 month-old, 73.9% (238/322) in 2~6-month old, and 73.6% (53/72) in less than 2-month old, respectively. Regional prevalence rate of PCMV infection in Korea showed 89.7% (70/78) in Chungchong, 79.8/% (83/104) in Cholla, 79.4% (143/180) in Kyonggi, 79.3% (42/53) in Gyongsang, 50% (15/30) in Kangwon, and 38.9% (14/36) in Cheju area, respectively. In the sera collected in 1991, seropositive rate was appeared as 90.2% (37/41). From 1992 to 1997 except 1994, the average infection rate to PCMV was 77.5%. Consequently, these results confirmed that PCMV in Korean piggeries was introduced at least before the year of 1991. More importantly, PCMV infection has been prevailing nation-wide in pig herds in Korea.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.19
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• pp.8177-8182
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• 2014

Background: Blocking angiogenesis by targeting vascular endothelial growth factor (VEGF) signaling pathway to inhibit tumor growth has proven to be successful in treating a variety of different metastatic tumor types, including kidney, colon, ovarian, and lung cancers, but its role in castration-resistant prostate cancer (CRPC) is still unknown. We here aimed to determine the efficacy and toxicities of anti-VEGF agents in patients with CRPC. Materials and Methods: The databases of PubMed, Web of Science and abstracts presented at the American Society of Clinical Oncology up to March 31, 2014 were searched for relevant articles. Pooled estimates of the objective response rate (ORR) and prostate-specific antigen (PSA) response rate (decline ${\geq}50%$) were calculated using the Comprehensive Meta-Analysis (version 2.2.064) software. Median weighted progression-free survival (PFS) and overall survival (OS) time for anti-VEGF monotherapy and anti-VEGF-based doublets were compared by two-sided Student's t test. Results: A total of 3,841 patients from 19 prospective studies (4 randomized controlled trials and 15 prospective nonrandomized cohort studies) were included for analysis. The pooled ORR was 12.4% with a higher response rate of 26.4% (95%CI, 13.6-44.9%) for anti-VEGF-based combinations vs. 6.7% (95%CI, 3.5-12.7%) for anti-VEGF alone (p=0.004). Similarly, the pooled PSA response rate was 32.4% with a higher PSA response rate of 52.8% (95%CI: 40.2-65.1%) for anti-VEGF-based combinations vs. 7.3% (95%CI, 3.6-14.2%) for anti-VEGF alone (p<0.001). Median PFS and OS were 6.9 and 22.1 months with weighted median PFS of 5.6 vs. 6.9 months (p<0.001) and weighted median OS of 13.1 vs. 22.1 months (p<0.001) for anti-VEGF monotherapy vs. anti-VEGF-based doublets. Conclusions: With available evidence, this pooled analysis indicates that anti-VEGF monotherapy has a modest effect in patients with CRPC, and clinical benefits gained from anti-VEGF-based doublets appear greater than anti-VEGF monotherapy.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.289-299
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• 1993

The polypeptide patterns of cellular and follicular components were analysed by SDS-PAGE and two dimensional(2-D)electrophoresis combined with isoelectric focusing (IEF) to establish protein profiles in each of the components in porcine follicles. Oocyte-cumulus complexes were cultured in M16+FCS+Gn at 39 in an atmosphere of 5% CO$_2$, in air for 35 h. At the end of the culture, the zona-free oocyte, ZP alone and cumulus cells were prepared and analysed either on 10% SDS-PAGE for the protein profile at the first dimensional gel or 2-D protein pattern. The amounts of each samples were determined for the visualization with Coomasie brilliant blue (CBB) or silver staining, thus giving useful information for the identification of specific proteins in the components or appropriate amount of samples for proper visualization. Oocyte showed 25 and 114 kd major protein band. Other minor components were additionally visualized with CBB on the same gel after silver staining procedure. Cumulus cells also showed specific proteins which is not present in the oocytes. The number of cumulus cell was proper to give major bands with CBB and additional minor bands with silver staining. To establish the degree of contamination from the remnant of the corona radiata to the ZP, zonae were differently prepared or analysed by SDS-PAGE.The preparation of the ZP in this study did not showed any contamination judged by the protein profile of the components. Also follicular fluid showed its specific protein profile without any significant differences among the different sizes of follicles. The established protein profile of each follicular component should be helpful for the identification and elimination of contaminated components, i. e., antigen preparation or immunological studies. The results also suggest that the preparation of each components in the study was appropriate and can be used for a further sensitive biochemical analysis in mammalian oocytes and early embryos.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.263-268
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• 2005

Porcine epidemic diarrhea virus (PEDV) causes acute enteritis in pigs of all ages and is often fatal for neonates. In order to develop sweetpotato plants expressing PEDV antigen, we constructed the vector expressing spike gene of PEDV under the control of sweetpotato sporamin promoter or constitutive CaMV 35S promoter. The spike protein region of PEDV was synthesized by PCR and linked to each promoter, Transgenic sweetpotato [Ipomoea batatas (L.) Lam. cv. Yulmi] plants were developed from embryogenic calli following Agrobacterium tumefaciens-mediated transformation. The co-cultured embryogenic calli transferred to selective MS medium containing 1 mg/L 2,4-D, 100 mg/L kanamycin, and 400 mg/L claforan. These embryogenic calli were subcultured to the same selection medium at 3 weeks interval. Kanamycin-resistant calli transferred to hormone-free MS medium with kanamycin gave rise to somatic embryos and then converted into plantlets in the same medium. Southern blot analysis confirmed that the spike gene of PEDV was inserted into the genome of the sweetpotato plants. RT-PCR revealed that the spike gene of PEDV was highly expressed in transgenic sweetpotato plants.