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Characteristics of β-casein Gene using the PCR Technique in Korean Native Goat (PCR 기법을 이용한 한국재래산양 β-casein 유전자의 특성)

  • Kim, Ji-Ae;Ryoo, Seung-Heui;Yu, Sung-Lan;Lee, Jun-Heon;Seo, Gil-Woong;Kim, Sun-Kyun;Sang, Byung-Chan
    • Korean Journal of Agricultural Science
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    • v.29 no.2
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    • pp.43-52
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    • 2002
  • This study was performed to provide the basic data for preservation and improvement of genetic resources according to finding genetic construction obtained from analysis of genetic characteristics of $\beta$-casein gene in Korean Native goat and Saanen using the PCR-RFLP. This study confirmed the amplified products of 481bp fragments obtained from the amplification of $\beta$-casein loci by PCR. The $\beta$-casein AB genotype showed 481, 284 and 197bp, and $\beta$-casein BB genotype showed 284 and 197bp fragments in Korean Native goat and Saanen. The frequencies of $\beta$-casein genotype in Korean Native goat were 6.25 and 93.75% for AA and AB and the frequencies of $\beta$-casein genotype in Saanen were 57.14 and 42.86% for AA and AB types. The frequencies of $\beta$-casein A and B alleles were 0.031 and 0.969 in Korean Native goat and the frequencies of $\beta$-casein A and B alleles are 0.286 and 0.714 in Saanen, respectively. The nucleotide sequence of $\beta$-casein gene of Korean Native goat was 97.71% higher homology with 11 nucleotide sequences difference of that of goat reported in GeneBank (M90556). Therefore, this study of molecular genetic characteristics by the analysis of genetic polymorphism and sequencing for $\beta$-casein gene should be used as basic and applying data for preservation and improvement of genetic resources in Korean Native goat breeding.

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Detection of Megalocytivirus in shellfish using PCR with various DNA extraction methods (다양한 PCR용 DNA 추출법에 의한 패류 내 Megalocytivirus의 검출)

  • Kim, Jin-Woo;Cho, Mi-Young;Jin, Ji-Woong;Kim, Ki-Hong;Jeong, Hyun-Do;Kim, Kwang-Il
    • Journal of fish pathology
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    • v.24 no.2
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    • pp.65-73
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    • 2011
  • In analysis of DNA viruses from the contaminated shellfish using PCR, preparation method of template DNA is an important factor to get enough copy number of viruses. In this study, we evaluated the efficiency of PCR template of Megalocytivirus (sT50mg-D) DNA obtained from 50 mg digestive gland homogenate of oyster using commercial method, and compared with that obtained from 5 g of the same tissues (T5g-D) after PEG precipitation procedures of virus. Both templates DNA suspended in the same volume of distilled water showed positive results by primary PCR with 35 cycles, and the presence of Megalocytivirus was confirmed in oysters collected from cultured farms in Korea. Moreover, PCR with sT50mg-D allowed us to discriminate the contaminated oyster individually, that can not be done in PCR with T5g-D prepared from the mixture of three different individual oyster to get 5 g digestive gland homogenate. In quantitative analysis with real time PCR, Megalocytivirus concentrations in 50 ${\mu}l$ templates prepared using 0.5~50 mg of one positive sample were appeared in the range 6.14E+00~1.2E+02/${\mu}l$. We were not able to get positive result using template DNA contained less than 6.14E+00 copies. Consequently, 2-step PCR performed with DNA extracts from oyster homogenate of small amount (sT50mg-D) i) was enough to detect the contaminated Megalocytivirus in shellfish, ii) allowed us to do the analysis for individual shellfish rather than mixture of several shellfish and iii) showed the presence of Megalocytivirus in oyster from Korea.

ANALYSIS OF DIFFERENTIAL GENE EXPRESSION IN NORMAL, CYST AND AMELOBLASTOMA CELLS (정상, 낭종 및 법랑아세포종 세포에서의 유전자 발현 차이 분석)

  • Yang, Cheol-Hee;Baik, Byeong-Ju;Yang, Yeon-Mi;Kim, Jae-Gon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.32 no.1
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    • pp.75-88
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    • 2005
  • Ameloblastoma is the most commonly occurring odontogenic tumor in oral cavity. Although most are benign epithelial neoplasm, they are generally considered to be locally aggressive and destructive, exhibiting a high rate of recurrence. The biological behavior of this neoplasm is a slowly growing, locally invasive tumor without metastasis, therefore malignant neoplasm, changed its histological appearance to carcinoma or showed distant metastasis, is only defined clinically. In this study, we identified the differentially expressed genes(DEGs) in stages under benign or malignant ameloblastoma compared with normal patient using ordered differential display(ODD) reverse transcription(RT)-PCR and $GeneFishing^{TM}$ technology. ODD RT-PCR is rather effective when the investigation of samples containing very small amounts of total RNA must be accomplished. ODD RT-PCR used the means of amplification with anchored T-primer and adaptor specific primer. bearing definite two bases at their 3' ends and so this method could display differential 3'-expressed sequence taqs(ESTs) patterns without using full-length cDNAs. Compared with standard differential display, ODD RT-PCR is more simple and have enough sensitivity to search for molecular markers by comparing gene expression profiles, However, this method required much effort and skill to perform. $GeneFishing^{TM}$ modified from DD-PCR is an improved method for detecting differentially expressed genes in two or more related samples. This two step RT-PCR method uses a constant reverse primer(anchor ACP-T) to prime the RT reaction and arbitrary primer pairs(annealing control primers, ACPs) during PCR. Because of high annealing specificity of ACPs than ODD RT-PCR, the application of $GeneFishing^{TM}$ to DEG discovery generates reproducible, authentic, and long(100bp to 2kb) PCR products that are detectable on agarose gels. Consequently, various DEGs observed differential expression levels on agarose gels were isolated from normal, benign, and malignant tissues using these methods. The expression patterns of the some isolated DEGs through ODD RT-PCR and $GeneFishing^{TM}$ were confirmed by Northern blot analysis and RT-PCR. The results showed that these identified DEGs were implicated in ameloblastoma neoplasm processes. Therefore, the identified DEGs will be further studied in order to be applied in candidate selection for marker as an early diagnosis during ameloblastoma neoplasm processes.

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Method Development of Ellagic Acid as Marker Compound for Standardization of Gochang Bokbunja (Rubus coreanus Miquel) as Functional Ingredient (고창 복분자의 기능성원료 표준화를 위한 지표성분으로서 Ellagic Acid의 분석법 개발)

  • Kim, Yunjeong;Han, Song-Hee;Jeon, Ji-Yeong;Hwang, Minho;Im, Yong-Jin;Chae, Soo-Wan;Kim, Min-Gul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.11
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    • pp.1554-1558
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    • 2012
  • Method development and validation of ellagic acid for the standardization of Gochang Bokbunja as a functional ingredient and health food were accomplished. A Symmetry$^{(R)}$ (C18, $4.6{\times}250mm$, $5.0{\mu}m$) column was used with a gradient elution system of 1% formic acid in water and acetonitrile. This method was validated according to specificity, linearity, accuracy, precision test, and recovery test. Specificity was confirmed with identical retention time, and calibration curves of ellagic acid showed good linear regression ($R^2$ >0.9996). Relative standard deviations (RSD) of data from the intra- and inter-day experiments were less than 2.28% and 2.84%, except in the low limit of quality control (LLOQ, $1{\mu}g/mL$) sample. The results of the recovery test were from 89.17% to 97.92% with RSD values from 0.05 to 0.14%. Therefore, we performed analysis of ellagic acid as a marker compound in Gochang Bokbunja extracts. The amount of ellagic acid in Gochang Bukbunja was about $1.918{\mu}g/mg$ (0.192%) in the three times analysis, and RSD was less than 2.36% by the validated method. These results suggest that the developed HPLC method is simple, efficient, and could contribute to the quality control of Gochang Bokbunja extract as a functional ingredient.

Comparison of Two Rapid Antigen Detection Tests for Diagnosis of Group A Streptococcal Pharyngotonsillitis (A군 연쇄구균성 인두편도염 진단을 위한 두 종류 신속항원검사법의 비교)

  • Song, Seung Kyu;Hong, Mi Ae;Oh, Kyung Chang;Ahn, Seung In;Tae, Mi Hyon;Shin, Hye Jung;Chang, Jin Keun;Cha, Sung Ho
    • Clinical and Experimental Pediatrics
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    • v.45 no.8
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    • pp.973-979
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    • 2002
  • Purpose : Recently, a number of rapid antigen detection tests have been available to diagnose group A streptococcal pharyngotonsillitis. The purpose of this study was to determine the sensitivity, specificity and consistency of the two rapid antigen detection tests. Methods : Among the patients who visited our clinic from November 2001 to February 2002, 61 patients who had clinical findings of pharyngeal erythema or edema, pharyngeal exudates and soft palatine petechiae were enrolled in our study. A total of 61 patients were tested with rapid antigen detection tests and throat culture. BD $LINK2^{TM}$ Strep A(Becton, Dickinson & Company, U.S.A.) and $QuickVue^{(R)}$ $In-Line^{TM}$(Quidel Corporation, U.S.A.) were selected for rapid antigen detection tests. Results : Of the 61 patients tested, 22 patients were confirmed as group A streptococcal pharyngotonsillitis by throat culture. The BD $LINK2^{TM}$ Strep A had a sensitivity of 81.8% and a specificity 89.7%. The positive and negative predictive values were 81.8% and 89.7%, respectively. The $QuickVue^{(R)}$ $In-Line^{TM}$ had a sensitivity of 77.3% and a specificity of 100%. The positive and negative predictive values were 100% and 88.6%, respectively. The kappa values of BD $LINK2^{TM}$ Strep A and $QuickVue^{(R)}$ $In-Line^{TM}$ were 0.72 and 0.81, respectively. Conclusion : In addition to high sensitivity, specificity and consistency, both kits are easy to use and simple to interpret, and therefore have the potential to be used with backup throat culture for diagnosis of acute pharyngotonsillitis.

Antioxidant and Cytoprotective Effects of Socheongja and Socheong 2, Korean Black Seed Coat Soybean Varieties, against Hydrogen Peroxide-induced Oxidative Damage in HaCaT Human Skin Keratinocytes (HaCaT 인간 피부 각질세포에서 과산화수소 유도 산화 손상에 대한 소청자 및 소총2호의 항산화 및 세포보호 효능)

  • Choi, Eun Ok;Kwon, Da Hye;Hwang, Hye-Jin;Kim, Kook Jin;Lee, Dong Hee;Choi, Yung Hyun
    • Journal of Life Science
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    • v.28 no.4
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    • pp.454-464
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    • 2018
  • Black soybeans are used as food sources as well as for traditional medicines because they contain an abundance of natural phenolic compounds. In this study, total phenolic contents (TPCs) of Korean black seed coat soybean varieties Socheongja (SCJ), Socheong 2 (SC2) and Cheongja 2 (CJ2) as well as their antioxidant capacities were investigated. Among them, TPCs were abundantly present in the order of CJ2$H_2O_2$-stimulated HaCaT human keratinocytes. Our results revealed that treatment with SCJ and SC2 prior to $H_2O_2$ exposure significantly increases the viability of HaCaT cells, indicating that the exposure of HaCaT cells to SCJ and SC2 conferred a protective effect against oxidative stress. SCJ and SC2 also effectively inhibited $H_2O_2$-induced apoptotic cell death through the blocking of mitochondrial dysfunction. SCJ and SC2 also attenuated the phosphorylation of Histone H2AX. Furthermore, they effectively induced the levels of thioredoxin reductase (TrxR) 1, a potent antioxidant enzyme, which is associated with the induction of nuclear transcription factor erythroid-2-like factor 2 (Nrf2); however, the protective effects of SCJ and SC2 were significantly reversed by Auranofin, a TrxR inhibitor. These results indicate that they have protective activity through the blocking of cellular damage related to oxidative stress via the Nrf2 signaling pathway. In conclusion, our study indicated that SCJ and SC2 might potentially serve as novel agents for the treatment and prevention of skin disorders caused by oxidative stress.

Development of a predictive model describing the growth of Staphylococcus aureus in processed meat product galbitang (식육추출가공품 중 갈비탕에서의 Staphylococcus aureus 성장예측모델 개발)

  • Son, Na-Ry;Kim, An-Na;Choi, Won-Seok;Yoon, Sang-Hyun;Suh, Soo-Hwan;Joo, In-Sun;Kim, Soon-Han;Kwak, Hyo-Sun;Cho, Joon-Il
    • Korean Journal of Food Science and Technology
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    • v.49 no.3
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    • pp.274-278
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    • 2017
  • In this study, predictive mathematical models were developed to estimate the kinetics of Staphylococcus aureus growth in processed meat product galbitang. Processed meat product galbitang was inoculated with 0.1 mL of S. aureus culture and stored at 4, 10, 20, $37^{\circ}C$. The ${\mu}_{max}$ (maximum specific growth rate) and LPD (lag phase duration) values were calculated. The primary model was used to develop a response surface secondary model. The growth parameters were analyzed using the square root model as a function of storage temperature. The developed model was confirmed by calculating RMSE (Root Mean Square Error) values as statistic parameters. The LPD decreased, but ${\mu}_{max}$ increased with an increase in the storage temperature. At 4, 10, 20 and $37^{\circ}C$, $R^2$ was 0.99, 0.98, 0.99 and 0.99, respectively; RMSE was 0.39. The developed predictive growth model can be used to predict the risk of S. aureus contamination in processed meat product galbitang; hence, it has potential as an input model for the risk assessment.

Serologic follow-up Study in neurocysticercosis patients by ELISA after praziquantel treatment (프라지콴텔 치료후 효소면역측정법에 의한 뇌 유구낭미충증 환자의 혈청학적 추적검사)

  • Cho, Seung-Yull;Kim, Suk-Il;Kang, Shin-Yong
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.159-170
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    • 1986
  • A total of 69 patients of confirmed neurocysticercosis was followed serologically by ELISA up to 22 months after praziquantel treatment. The intervals and numbers of follow-up were variable by patient. Serially collected samples of serum and CSF were examined simultaneously for their specific IgG antibody levels by ELISA, using cystic fluid, saline extracts of bladder wall and scolex as antigen. Within 4 months after praziquantel treatment, the antibody levels were elevated temporarily in both serum and CSF in most patients. In some cases antibody levels exhibited steady declining tendency after the treatment. Concomitant administration of dexamethasone appeared to suppress the elevation of antibody levels. The rate of mean absorbance of antibody changed more in serum than in CSF. The rate of elevation was greater in antibodies to parenchymal antigens than that to cystic fluid, but absolute difference of antibody levels was greater in antibody to cystic fluid. Previously negative samples for IgG antibody may become positive after the praziquantel treatment, which could be used as a complementary tool (provocation test) in serodiagnosis. One month was considered to be sufficient interval for the follow-up test for that purpose. In the follow-up of up to 22 months, only few cases of chronic neurocysticercosis showed declining tendency of IgG antibody levels below negative range. During acute encephalitic attacks in chronic patients, IgG antibody to parenchymal antigen were elevated in CSF temporarily. These results indicated that serologic follow-up of every year was recommendable to differentiate the cured patients from chronic patients with slowly calcifying lesions.

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Prevalence and Risk Factors Associated with Esophagitis in Children with Abdominal Pain (복통이 있는 소아청소년에서 식도염의 유병률과 위험인자)

  • Kwon, Hyun-Jung;Yi, Dae-Yong;Ryoo, Eell;Cho, Kang-Ho;Son, Dong-Woo;Tcha, Han
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.11 no.2
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    • pp.103-109
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    • 2008
  • Purpose: Children with esophagitis express a variety of nonspecific symptoms and signs depending on their age, and diagnosis is limited because gastrointestinal endoscopy (GFS) and biopsy are difficult to perform. The aim of this study was to examine the prevalence of esophagitis in children with upper abdominal pain, to determine the necessity of esophageal biopsy, and to evaluate the associated risk factors. Methods: We reviewed 266 pediatric patients with upper abdominal pain who underwent history-taking, physical examination, and GFS with esophageal and gastric biopsies between January 2006 and December 2007. Esophagitis was confirmed on biopsy. We analyzed the risk factors for histologic esophagitis and the necessity of esophageal biopsy. Results: The prevalence of esophagitis was 19.9% (53/266 patients). The sensitivity and specificity of endoscopic diagnosis were 41.5% and 77%. Of 53 patients with histologic esophagitis, reflux esophagitis was seen in 50 patients, eosinophilic esophagitis was seen in 2 patients, and esophageal candidiasis was seen in 1 patient. Vomiting was a significant factor in patients under 8 yr of age (p<0.05). H. pylori infection was documented in 41.5% of patients with histologic esophagitis, compared with 58.5% of patients not infected with H. pylori (p<0.05). The possibility of histologic esophagitis was higher in patients with H. pylori infection (OR 2.5, 95% CI 1.2544 to 4.8286) and in those who visited in the spring (OR 2.5, 95% CI 1.2544 to 4.8286). Conclusion: We believe esophageal tissue biopsy should be performed in pediatric patients with upper gastrointestinal symptoms who are undergoing GFS and stomach tissue biopsy, especially preschoolers and H. pylori-infected children in the spring.

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Study on Ecological Restoration of Endangered Species in Abandoned Paddy of Korea and Management Plan for its Habitat (한국의 묵논에서 멸종위기식물의 복원생태학적 연구 및 서식지 관리방안)

  • Lee, Soo-In;Lee, Eung-Pill;Hong, Young-Sik;Kim, Eui-Joo;Lee, Seung-Yeon;Park, Jae-Hoon;Jang, Rae-Ha;You, Young-Han
    • Journal of Wetlands Research
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    • v.22 no.2
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    • pp.81-91
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    • 2020
  • As part of method for ecologically utilizing abandoned paddy, potential of the abandoned paddy as a target site for ex-situ conservation of 9 endangered species of different life form was confirmed by considering the nature of rice paddy. In order to create Dum-bung, a component of traditional Korean rice paddy, a part of the abandoned paddy was modified to change the water environment. The seeds, asexual reproductive and sexually reproductive individuals of each endangerd species were transplanted into the abandoned paddy to observe the survival rate and phenological response for 1 year, and then monitored for 2 years. As a result, Hydrophyte 4 taxa (Euryale ferox, Saururus chinensis, Dysophylla yatabeana, Menyanthes trifoliata), Geophyte taxa 1 (Epilobium hirsutum), and Hemicryptophyte taxa 1 (Cicuta virosa) could be introduced into the abandoned paddy. In particular, Euryale ferox, Dysophylla yatabeana, and Menyanthes trifoliata should be introduced into Dum-bung, and Saururus chinensis, Epilobium hirsutum, and Cicuta virosa should be introduced into paddy wetland. Growth of Euryale ferox and Brasenia schreberi was inhibited by herbaceous species, and the growth of Epilobium hirsutum was inhibited by herbivores. Therefore, in order to help efficient settlement of endangered plants introduced in abandoned paddy, it is necessary to remove herbs that inhibit growth and to manage herbivores. In addition, it is necessary to prevent the collapse of paddy bank by planting on the paddy field trees or herbaceous forming vegetation mat. When using abandoned paddy ecologically, it is effective to diversify the moisture environment by creating a Dum-bung to increase biodiversity.