• Title/Summary/Keyword: ITS marker

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Clinical relevance of Lgr5 expression in colorectal cancer patients

  • Kim, Young Joo;Kang, Dong Hyun;Song, Geum Jong;Ahn, Tae Sung;Son, Myoung Won;Lee, Moon Soo;Baek, Moo-Jun
    • Korean Journal of Clinical Oncology
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    • v.14 no.2
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    • pp.76-82
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    • 2018
  • Purpose: Lgr5 is a well-known stem cell marker in colorectal cancer (CRC). This retrospective study evaluated the expressions of Lgr5 in CRC specimens, and examined whether these expressions were associated with survival outcomes. Methods: We used immunohistochemistry to retrospectively examine expressions of Lgr5 in paraffin-embedded specimens from 337 patients with CRC between January 2009 and December 2013. All clinicopathologic data were collected by retrospective review based on medical records. The correlation between its expression and clinicopathological data as well as clinical outcomes of patients was analyzed. Results: Low expression and high expression of Lgr5 in 337 patients were 175 (51.9%) and 162 (48.1%), respectively. There was no statistically significant difference in the association of Lgr5 expression with clinicopathologic factors (age, tumor location, lymphatic invasion, vascular invasion, perineural invasion, TNM stage, and differentiation). In the survival analysis, the high expression group of Lgr5 showed a better prognosis than the low expression group in disease-free survival (P=0.044). However, overall survival was not significantly different between the two groups (P=0.087). In multivariate analysis, we found that high expression of Lgr5 was independent prognostic factor for tumor relapse (hazard ratio, 0.601; 95% confidence interval, 0.388-0.929; P=0.022). Conclusion: In present study, high expression of Lgr5 is an independent predictor of favorable prognosis in patients with CRC. So, further well designed, prospective, large scale studies are needed to examine the value of Lgr5 as a prognostic biomarker for CRC.

Genotyping of avian pathogenic Escherichia coli by DNA fragment analysis for the differences in simple sequence repeats

  • Han, Mi Na;Byeon, Hyeon Seop;Han, Seong Tae;Jang, Rae Hoon;Kim, Chang Seop;Choi, Seok Hwa
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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    • pp.257-262
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    • 2018
  • Avian pathogenic E. coli (APEC) causes severe economic losses in the poultry farms, due to systemic infections leading to lethal colisepticemia. It causes a variety of diseases from air sac infection to systemic spread leading to septicemia. Secondary infection contains opportunistic infections due to immunosuppression disease. Collibacillosis causes the great problems in the poultry industry in Korea. Thus, it is necessary to identify and classify the characteristics of E. coli isolate of chicken origin to confirm the diversity of symptoms and whether they are transmitted among the farms. Fragment analysis is identify the difference in the number of Variable-Number Tandem-Repeats (VNTRs) for genotyping. VNTRs have repeating structure (Microsatellite, Short tandem repeats; STR, Simple sequence repeats; SSR) in the chromosome. This region can be used as a genetic marker because of its high mutation rate. And various lengths of the amplified DNA fragment cause the difference in the number of repetition of the DNA specific site. The number of repetition sequences indicates the separated size of fragments, so the each fragments can be distinguished by specific samples. The results of the sample show that there is no difference in six microsatellite loci (yjiD, aidB, molR_1, ftsZ, b1668, yibA). There are differences among the farms in relation of the number of repetitions of other six microsatellite loci (ycgW, yaiN, yiaB, mhpR, b0829, caiF). Four (ycgW, yiaB, b0829, caiF) of these six microsatellite loci show statistically significant differences (P<0.05). It means that the analysis using four microsatellite loci including ycgW, yiaB, b0829, and caiF can confirm among the farms. Five E. coli samples in one farm have same SSR repetition at all markers. But, there are significant differences from other farms at Four (ycgW, yiaB, b0829, caiF) microsatellite loci. These results emphasize again that the four microsatellite loci makes a difference in the amplified DNA fragments, enabling it to be used for E. coli genotyping.

Race- and Isolate-specific Molecular Marker Development through Genome-Realignment Enables Detection of Korean Plasmodiophora brassicae Isolates, Causal agents of Clubroot Disease

  • Jeong, Ji -Yun;Robin, Arif Hasan Khan;Natarajan, Sathishkumar;Laila, Rawnak;Kim, Hoy-Taek;Park, Jong-In;Nou, Ill-Sup
    • The Plant Pathology Journal
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    • v.34 no.6
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    • pp.506-513
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    • 2018
  • Clubroot is one of the most economically important diseases of the Brassicaceae family. Clubroot disease is caused by the obligate parasite Plasmodiophora brassicae, which is difficult to study because it is nonculturable in the laboratory and its races are genetically variable worldwide. In Korea, there are at least five races that belongs to four pathotype groups. A recent study conducted in Korea attempted to develop molecular markers based on ribosomal DNA polymorphism to detect P. brassicae isolates, but none of those markers was either race-specific or pathotype-specific. Our current study aimed to develop race- and isolate-specific markers by exploiting genomic sequence variations. A total of 119 markers were developed based on unique variation exists in genomic sequences of each of the races. Only 12 markers were able to detect P. brassicae strains of each isolate or race. Ycheon14 markers was specific to isolates of race 2, Yeoncheon and Hoengseong. Ycheon9 and Ycheon10 markers were specific to Yeoncheon isolate (race 2, pathotype 3), ZJ1-3, ZJ1-4 and ZJ1-5 markers were specific to Haenam2 (race 4) isolate, ZJ1-35, ZJ1-40, ZJ1-41 and ZJ1-49 markers were specific to Hoengseong isolate and ZJ1-56 and ZJ1-64 markers were specific to Pyeongchang isolate (race 4, pathotype 3). The PCR-based sequence characterized amplified region (SCAR) markers developed in this study are able to detect five Korean isolates of P. brassicae. These markers can be utilized in identifying four Korean P. brassicae isolates from different regions. Additional effort is required to develop race- and isolate-specific markers for the remaining Korean isolates.

Establishment and validation of an analytical method for quality control of health functional foods derived from Agastache rugosa

  • Park, Keunbae;Jung, Dasom;Jin, Yan;Kim, Jin Hak;Geum, Jeong Ho;Lee, Jeongmi
    • Analytical Science and Technology
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    • v.32 no.3
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    • pp.96-104
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    • 2019
  • Agastache rugosa, known as Korean mint, is a medicinal plant with many beneficial health effects. In this study, a simple and reliable HPLC-UV method was proposed for the quantification of rosmarinic acid (RA) in the aqueous extracts of A. rugosa. RA was selected as a quantification marker due to its easiness in procurement and analysis. The developed method involved chromatographic separation on a $C_{18}$ column ($250{\times}4.6mm$, $5{\mu}m$) at room temperature. The mobile phase consisted of water and acetonitrile both containing 2 % acetic acid and was run at a flow rate of $1mL\;min^{-1}$. The method was validated for specificity, linearity, precision, and accuracy. It was specific to RA and linear in the range of $50-300{\mu}g\;mL^{-1}$ ($r^2=0.9994$). Intra-day, inter-day, and inter-analyst precisions were ${\leq}0.91%\;RSD$, ${\leq}1.40%\;RSD$, and 1.94 % RSD, respectively. Accuracy was 93.3-95.9 % (${\leq}1.21%\;RSD$). The method could be applied to three batches of bulk samples and three batches of lab scale samples, which were found to be $0.64({\pm}0.04)mg\;g^{-1}$ and $0.48({\pm}0.02)mg\;g^{-1}$ for the dried raw materials of A. rugosa. The results show that the proposed method can be used as a readily applicable method for QC of health functional foods containing the aqueous extracts of A. rugosa.

Characteristics as Pulp and Papermaking Resources of Paper-mulberry (B. kazinoki) woods in Jeolla-do Region with Identically Genetic Marker (유전형질이 동일한 전라도지역 닥나무의 펄프제지 자원으로써의 특성)

  • Jo, Ah Hyeon;Go, In Hee;Jang, Kyung Ju;Park, Kyu Tae;Park, Sun Mi;Park, Seon Joo;Jeong, Seon Hwa
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.67-67
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    • 2019
  • 뽕나무과(Moraceae) 닥나무속(Broussonetia)의 품종에 대한 국내 연구동향으로는 잎의 성상이나 암꽃의 포길이, 엽신의 폭과 같은 수목학 분수학적 관점으로 특징을 파악하기 때문에 구분이 어려운 실정이다. 또한 닥나무(Broussonetia kazinoki Siebold)와 꾸지나무[Broussonetia papyrifera (L.) $L^{\prime}H{\acute{e}}r.$ ex Vent.]의 수목학적 특징이 유사하여 오동정의 사례가 발생하기도 한다. 닥나무의 향명은 다양하게 불리지만 그 분류기준은 명확하지 않다. 본 연구에서는 총 4개 마커(ITS, matK, trnL-F, ndhF)의 염기서열 분석과 분자계통학적 분류를 통해 닥나무와 꾸지나무의 교잡종으로 확인된 전라도지역 닥나무 8개체에 대하여 펄프제지 자원으로써의 특성을 확인하였다. 8개체 인피섬유의 형태학적 특징인 섬유장, 섬유폭, 섬유 내강(Lumen)폭을 통해 Runkel ratio, Slenderness ratio, Flexibility coefficient, Rigidity coefficient를 도출하고 종이의 기계적 강도, 내절도와의 상관관계, 섬유와 섬유간의 결합력 등을 예측하였다. 상기와 같은 연구결과는 유전형질이 동일한 전라도지역 닥나무의 형태학적 특성을 통해 펄프제지 공정에 영향을 미칠 수 있는 인자들을 분석 하였다. 이러한 특성에 따라 다양한 품질의 종이를 제조 할 수 있는 기초 자료로 제공될 것이다.

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Qualitative and quantitative determination of oleanolic acid in a scalp tonic products by HPLC using response surface methodology for extraction optimization

  • Cai, Lin Xi;Cho, Chong Woon;Zhao, Yan;Kang, Jong Seong;Kim, Kyung Tae;Jung, Sang-Hun
    • Analytical Science and Technology
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    • v.32 no.2
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    • pp.48-55
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    • 2019
  • The simple and effective analytical method for the quality control of a novel scalp tonic formulation has been developed and optimized in terms of HPLC conditions and sample preparation method, meanwhile, the optimization of preparation condition was using response surface methodology (RSM) based on central composite design (CCD). Oleanolic acid was selected as marker compound because of its bioactivities for alopecia therapy. The developed analytical method and extraction condition were successfully qualified. Coefficient of determination ($r^2$) for the calibration was 0.9997 with a line passing through the origin point in the range of 0.1-100 mg/mL. The limit of detection (LOD) and the limit of quantitation (LOQ) were 17.5 ng/mL and 55.0 ng/mL, respectively. The intra-day and inter-day precision of the method were 0.5-1.4 % and 0.7-1.8 % in relative standard deviation, respectively, while those accuracy were 99.5-100.9 % and 100.0-102.2 %, respectively. The repeatability of oleanolic acid in samples ranged of 0.3-1.9 % based on peak area and 0.3-0.7 % for retention time. Recoveries from samples were 95.0-99.4 % with lower than 1.8 % in relative standard deviation. Overall, the developed analytical method will be used for quality control of this commercial scalp tonic products successfully.

The changes of stresses and ecdysteroid biosynthesis gene expression levels in Kynurenine 3-monooxygenase mutant Bombyx mori

  • Jeong, Chan Young;Lee, Chang Hoon;Kim, Su Bae;Kang, Sang Kuk;Ju, Wan-Taek;Kim, Seong-Wan;Kim, Nam-Suk;Kim, Kee Young;Park, Jong Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.43 no.1
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    • pp.29-36
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    • 2021
  • Silkworms have long been bred with human history to produce silk. It has been with humans for longer than other industrial insects, and the silkworm variety has been continuously improved. Silkworms have been developed into the optimal form for producing high quality silk and pupae. Recently, the production of transgenic silkworms has further expanded the possibility of industrial value of silkworms. Kynurenine 3-monooxygenase (KMO), which is a flavin enzyme, is known for its involvement in ommochrome pigment synthesis. In the field of mammals, including humans, previous studies have revealed the function and role of KMO, which is an important enzyme for various immune responses and cell protection. However, in the case of insects, the function of KMO has only been studied to be involved in the formation of pigment, and accordingly, KMO is used exclusively on screening for generation of transgenic insects as a marker. In this study, using KMO-edited silkworms, it was intended to discover the novel functions and roles of KMO in silkworms by identifying changes in the expression of various genes associated with stress and growth. The changes were observed in expressions of genes regulating on stresses to survive and those on ecdysteroid hormone between wild-type (WT) silkworms and kmo mutant silkworms. The loss of KMO, in particular, decreased the expression of the shadow (sad) gene, one of the Halloween genes in the synthesis of ecdysteroid. In conclusion, these results suggest that silkworm KMO is responsible for potential functions regarding stress response and ecdysteroid synthesis.

Evaluation of Genetic Diversity and Population Structure Analysis among Germplasm of Agaricus bisporus by SSR Markers

  • An, Hyejin;Lee, Hwa-Yong;Shin, Hyeran;Bang, Jun Hyoung;Han, Seahee;Oh, Youn-Lee;Jang, Kab-Yeul;Cho, Hyunwoo;Hyun, Tae Kyung;Sung, Jwakyung;So, Yoon-Sup;Jo, Ick-Hyun;Chung, Jong-Wook
    • Mycobiology
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    • v.49 no.4
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    • pp.376-384
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    • 2021
  • Agaricus bisporus is a popular edible mushroom that is cultivated worldwide. Due to its secondary homothallic nature, cultivated A. bisporus strains have low genetic diversity, and breeding novel strains is challenging. The aim of this study was to investigate the genetic diversity and population structure of globally collected A. bisporus strains using simple sequence repeat (SSR) markers. Agaricus bisporus strains were divided based on genetic distance-based groups and model-based subpopulations. The major allele frequency (MAF), number of genotypes (NG), number of alleles (NA), observed heterozygosity (HO), expected heterozygosity (HE), and polymorphic information content (PIC) were calculated, and genetic distance, population structure, genetic differentiation, and Hardy-Weinberg equilibrium (HWE) were assessed. Strains were divided into two groups by distance-based analysis and into three subpopulations by model-based analysis. Strains in subpopulations POP A and POP B were included in Group I, and strains in subpopulation POP C were included in Group II. Genetic differentiation between strains was 99%. Marker AB-gSSR-1057 in Group II and subpopulation POP C was confirmed to be in HWE. These results will enhance A. bisporus breeding programs and support the protection of genetic resources.

Evaluation of the acute toxicity of theoredoxin (TRX) transgenic soybean to Daphnia magna

  • Oh, Sung-Dug;Min, Seok-Ki;Kim, Jae Kwang;Park, Jung-Ho;Kim, Chang-Gi;Park, Soo Yun
    • Korean Journal of Agricultural Science
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    • v.47 no.4
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    • pp.791-802
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    • 2020
  • Theoredoxin (TRX) transgenic soybeans were developed using the human Theoredoxin gene under the control of the ��-conglycinin promoter with a selection marker, the phosphinothricin acetyltransferase (PAT) gene. This study was done to assess the acute toxicity of a genetically modified (GM) soybean using the fresh water planktonic crustacean Daphnia magna. The acute toxicity effect of the TRX soybean and non-GM soybean (Gwangan) on D. magna was investigated at different concentrations (0, 156, 313, 625, 1,250, 2,500, and 5,000 mg·L-1). The TRX soybean used for the test was confirmed to express the TRX/PAT genes by PCR and enzyme-linked immunosorbent assay (ELISA). D. magna feeding tests showed no significant differences in the cumulative immobility or an abnormal response with either the TRX soybean or non-GM soybean. The feeding study showed a similar abnormal response and cumulative immobility of the D. magna between the TRX soybean and Gwangan treatments. Additionally, the 48 h-EC50 values for the TRX and Gwangan soybeans were 755.6 and 778 mg·L-1, respectively. The soybean NOEC (no observed effect concentration) value for D. magna was suggested to be 156 mg·L-1. These results suggest that there is no significant difference in toxicity to Daphnia magna between the TRX soybean and its non-GM counterpart.

Association between High Sensitivity C-Reactive Protein and Metabolic Syndrome in South Korea: A Gender-Specific Analysis (우리나라 성인의 고감도 C-반응성 단백과 대사증후군의 관련성: 성별 분석)

  • Shin, Eunyoung;Lee, Yongjae;Kim, Taehyun;Jung, Keum Ji;Chung, Woojin
    • Health Policy and Management
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    • v.31 no.2
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    • pp.158-172
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    • 2021
  • Background: Metabolic syndrome has been known as a risk of cardiovascular disease. Meanwhile, high sensitivity C-reactive protein (hs-CRP) is used as a predictor of cardiovascular disease. In this paper, we aimed to investigate the association between hs-CRP and metabolic syndrome. Method: A total of 7,633 were chosen as the study population from the 7th Korea National Health and Nutrition Examination Survey dataset (2016-2017). Our dependent variable was whether an individual had metabolic syndrome or not, and the independent variable of interest was hs-CRP which was categorized into three groups. The chi-square tests and hierarchical logistic regression analyses reflecting survey characteristics were conducted. All analyses were stratified by gender. Results: According to the adjusted model with all covariates, compared to individuals having the low risk of hs-CRP, those having its average risk were more likely to have metabolic syndrome in men (odds ratio [OR], 1.41; 95% confidence interval [CI], 1.12-1.76) and women (OR, 1.69; 95% CI, 1.33-2.16). Individuals having the high risk was not significantly different in men; however, they were more likely to have metabolic syndrome in women (OR, 2.03; 95% CI, 1.28-3.23). Conclusion: In an upcoming aging society, it is important to reduce the risk of metabolic syndrome to improve population health. This study suggests that hs-CRP may be used as a marker of the risk of metabolic syndrome in a gender-specific way, thereby contributing to enhancing awareness of the risk of metabolic syndrome among the general public.