• Title/Summary/Keyword: High-resolution melting analysis

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Development of HRM Markers Based on SNPs Identified from Next Generation Resequencing of Susceptible and Resistant Parents to Gummy Stem Blight in Watermelon (수박에서 덩굴마름병 감수성 및 저항성 양친에 대한 차세대 염기서열 재분석으로 탐색된 SNP 기반 HRM 분자표지 개발)

  • Lee, Eun Su;Kim, Jinhee;Hong, Jong Pil;Kim, Do-Sun;Kim, Minkyong;Huh, Yun-Chan;Back, Chang-Gi;Lee, Jundae;Lee, Hye-Eun
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.424-433
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    • 2018
  • Watermelon (Citrullus lanatus) is an economically important vegetable crop all over the world, which has functional compounds such as lycopene and citrulline. Gummy stem blight caused by Didymella bryoniae is one of the most devastative diseases in watermelon. Single nucleotide polymorphisms (SNPs), which are genetic variations occurring between individuals with respect to a single base, were often used to construct genetic linkage maps and develop molecular markers linked to a variety of horticultural traits and resistance to several diseases. In this study, we developed high-resolution melting (HRM) markers based on SNPs generated from NGS resequencing of two parents in watermelon. Plant materials were C. lanatus '920533' (female and susceptible parent), C. amarus 'PI 189225' (male and resistant parent), and their $F_1$ and $F_2$ progenies. A total of 13.6 Gbp ('920533') and 13.1 Gbp ('PI 189225') of genomic sequences were obtained using NGS analysis. A total of 6.09 million SNPs between '920533' and 'PI 189225' were detected, and 354,860 SNPs were identified as potential HRM primer sets. From these, a total of 330 primer sets for HRM analysis were designed. As a result, a total of 61 HRM markers that have polymorphic melting curves were developed. These HRM markers can be used for the construction of SNP-based linkage maps and for the analysis of quantitative trait loci (QTLs) related to gummy stem blight resistance.

Quantitative Analysis of Snow Particles Using a Multi-Angle Snowflake Camera in the Yeongdong Region (영동지역에서 눈결정 카메라를 활용한 눈결정의 정량 분석)

  • Kim, Su-Hyun;Ko, Dae-Hong;Seong, Dae-Kyung;Eun, Seung-Hee;Kim, Byung-Gon;Kim, Baek-Jo;Park, Chang-Geun;Cha, Ju-Wan
    • Atmosphere
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    • v.29 no.3
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    • pp.311-324
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    • 2019
  • We employed a Multi-Angle Snowflake Camera (MASC) to quantitatively analyze snow particles at the ground level in the Yeongdong region of Korea. The MASC captures high-resolution photographs of hydrometeors from three angles and simultaneously measures fallspeed. Based on snowflake images of the several episodes in 2017 and 2018, we derived statistics of size, aspect ratio, orientation, complexity, and fallspeed of snow crystals, which generally showed similar characteristics to the previous studies in other regions of the world. Dominant snow crystal habits of January 22, 2018 generated by northerly were melted aggregates when 850 hPa temperature was about $-6{\sim}-8^{\circ}C$. Average fallspeed of snow crystals was $1.0m\;s^{-1}$ though its size gradually increased as temperature decreased. Another snowfall event (March 8, 2018) was driven by the baroclinic instability as accompanied with a deep trough. Snow crystal habits were largely rimed aggregates (complexity ~1.8) and melting particles of dark images. Meanwhile, in the extreme snowfall event whose snow rate was greater than $10cm\;hr^{-1}$ on January 20, 2017, main snow crystals appeared to be heavily rimed particles with relatively smaller size when convective clouds developed vertically up to 9 km in association with tropopause folding. MASC also could successfully measure a decrease in snow crystal size and an increase in riming degree after AgI seeding at Daegwallyeong on March 14, 2017.

Mutation Analysis of KRAS and BRAF Genes in Metastatic Colorectal Cancer: a First Large Scale Study from Iran

  • koochak, Aghigh;Rakhshani, Nasser;Niya, Mohammad Hadi Karbalaie;Tameshkel, Fahimeh Safarnezhad;Sohrabi, Masoud Reza;Babaee, Mohammad Reza;Rezvani, Hamid;Bahar, Babak;Imanzade, Farid;Zamani, Farhad;Khonsari, Mohammad Reza;Ajdarkosh, Hossein;Hemmasi, Gholamreza
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.603-608
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    • 2016
  • Background: The investigation of mutation patterns in oncogenes potentially can make available a reliable mechanism for management and treatment decisions for patients with colorectal cancer (CRC). This study concerns the rate of KRAS and BRAF genes mutations in Iranian metastatic colorectal cancer (mCRC) patients, as well as associations of genotypes with clinicopathological features. Materials and Methods: A total of 1,000 mCRC specimens collected from 2008 to 2012 that referred to the Mehr Hospital and Partolab center, Tehran, Iran enrolled in this cross sectional study. Using HRM, Dxs Therascreen and Pyrosequencing methods, we analyzed the mutational status of KRAS and BRAF genes in these. Results: KRAS mutations were present in 33.6% cases (n=336). Of KRAS mutation positive cases, 85.1% were in codon 12 and 14.9% were in codon 13. The most frequent mutation at KRAS codon 12 was Gly12Asp; BRAF mutations were not found in any mCRC patients (n=242). In addition, we observed a strong correlation of KRAS mutations with some clinicopathological characteristics. Conclusions: KRAS mutations are frequent in mCRCs while presence of BRAF mutations in these patients is rare. Moreover, associations of KRAS genotypes with non-mucinous adenocarcinoma and depth of invasion (pT3) were remarkable.

Association of 8 Loci on Chromosome 8q24 with Prostate Carcinoma Risk in Northern Chinese Men

  • Zhao, Cheng-Xiao;Liu, Ming;Wang, Jian-Ye;Xu, Yong;Wei, Dong;Yang, Kuo;Yang, Ze
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6733-6738
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    • 2013
  • Multiple genetic studies have confirmed association of 8q24 variants with susceptibility to prostate cancer (PCa). As PCa risk SNPs may also influence disease outcome, we studied here eight 8q24 risk alleles, and evaluated their role in PCa clinical covariates in northern Chinese men. Blood samples and clinical information were collected from ethnically Chinese men from Northern China with histologically-confirmed PCa (n=289) and from age-matched normal controls (n=288). Eight 8q24 SNPs were genotyped by polymerase chain reaction-high- resolution melting analysis in 577 subjects. We examined the prevalence distribution of 8q24 risk alleles and analyzed the associations between the risk allele and PCa and clinical covariates to infer their impact on aggressive PCa. Three of the eight SNPs were associated with PCa risk in northern Chinese men, including rs16901966 (OR 1.31, 95% CI 1.01-1.70, p=0.042), rs1447295 (OR 1.47, 95% CI 1.09-1.98, p=0.011) and rs10090154 (OR 1.55, 95% CI 1.14-2.12, p=0.005). Haplotype analysis based association with the risk alleles revealed significant differences between cases and controls (OR 1.43, 95%CI 0.99-2.06, p=0.049). The risk alleles rs16901966, rs1447295 and rs10090154 were associated with age at diagnosis and tumor stage as compared with controls, while rs16901966 was associated with aggressive PCa (OR 1.43, 95% CI 1.01-2.03, p=0.042). The evidence for 8q24 SNPs with PCa risk in northern Chinese men showed rs16901966, rs1447295 and rs10090154 at 8q24 (region 1, region 2) to be strongly associated with PCa and clinical covariates. The three SNPs at 8q24 could be PCa susceptible genetic markers in northern Chinese men.

Development of Specific SNP Molecular Marker from Thistle in the DNA Sequences of Chloroplast TrnL-F and Matk Region Using HRM Analysis (엉겅퀴의 엽록체 TrnL-F와 Matk 영역 염기서열의 HRM 분석을 통한 특이적 SNP 분자마커의 개발)

  • Lee, Shin-Woo;Lee, Soo Jin;Kim, Yun-Hee
    • Journal of Life Science
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    • v.29 no.5
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    • pp.524-529
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    • 2019
  • Medicinal plants resources are becoming important assets since their usages have been expanded to the development of functional foods for human health, cosmetics and pharmaceutical industries. However, their phylogenetic origins and names are different from each country and quite often they are mixed each other resulting in the confusion for consumers. Particularly when they are very similar based on their morphological characteristics and distributed, it is extremely difficult to differentiate their origins even by specialists. Therefore, identification of each plant species is important for standardizing herbal medicine. Thistle is a medicinal and perennial plant. Obtaining information about the genetic diversity of plant populations is highly important for conservation and germplasm utilization. Although thistle is an important medicinal plant species registered in South Korea, no molecular markers are currently available to distinguish from other similar species from different countries. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific thistle species via high resolution melting (HRM) curve analyses. We performed molecular authentication of four different kinds of thistle species from different regions using DNA sequences in the trnL-F and matK chloroplast intergenic region. The SNP markers developed in this study are useful for rapidly identifying specific thistle species from different country.

8q24 rs4242382 Polymorphism is a Risk Factor for Prostate Cancer among Multi-Ethnic Populations: Evidence from Clinical Detection in China and a Meta-analysis

  • Zhao, Cheng-Xiao;Liu, Ming;Xu, Yong;Yang, Kuo;Wei, Dong;Shi, Xiao-Hong;Yang, Fan;Zhang, Yao-Guang;Wang, Xin;Liang, Si-Ying;Zhao, Fan;Zhang, Yu-Rong;Wang, Na-Na;Chen, Xin;Sun, Liang;Zhu, Xiao-Quan;Yuan, Hui-Ping;Zhu, Ling;Yang, Yi-Ge;Tang, Lei;Jiao, Hai-Yan;Huo, Zheng-Hao;Wang, Jian-Ye;Yang, Ze
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.19
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    • pp.8311-8317
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    • 2014
  • Background: Evidence supporting an association between the 8q24 rs4242382-A polymorphism and prostate cancer (PCa) risk has been reported in North American and Europe populations, though data from Asian populations remain limited. We therefore investigated this association by clinical detection in China, and meta-analysis in Asian, Caucasian and African-American populations. Materials and Methods: Blood samples and clinical information were collected from ethnically Chinese men from Northern China with histologically-confirmed PCa (n=335) and from age-matched normal controls (n=347). The 8q24 (rs4242382) gene polymorphism was genotyped by polymerase chain reaction-high-resolution melting analysis. We initially analyzed the associations between the risk allele and PCa and clinical covariates. A meta-analysis was then performed using genotyping data from a total of 1,793 PCa cases and 1,864 controls from our study and previously published studies in American and European populations, to determine the association between PCa and risk genotype. Results: The incidence of the risk allele was higher in PCa cases than controls (0.222 vs 0.140, $P=7.3{\times}10^{-5}$), suggesting that the 8q24 rs4242382-A polymorphism was associated with PCa risk in Chinese men. The genotypes in subjects were in accordance with a dominant genetic model (ORadj=2.03, 95%CI: 1.42-2.91, $Padj=1.1{\times}10^{-4}$). Presence of the risk allele rs4242382-A at 8q24 was also associated with clinical covariates including age at diagnosis ${\geq}65$ years, prostate specific antigen >10 ng/ml, Gleason score <8, tumor stage and aggressive PCa, compared with the non-risk genotype ($P=4.6{\times}10^{-5}-3.0{\times}10^{-2}$). Meta-analysis confirmed the association between 8q24 rs4242382-A polymorphism and PCa risk (OR=1.62, 95%CI: 1.39-1.88, $P=1.0{\times}10^{-5}$) across Asian, Caucasian and African American populations. Conclusions: The replicated data suggest that the 8q24 rs4242382-A variation might be associated with increased PCa susceptibility in Asian, Caucasian and African American populations. These results imply that this polymorphism may be a useful risk biomarker for PCa in multi-ethnic populations.

Analysis of Optical Characteristic Near the Cloud Base of Before Precipitation Over the Yeongdong Region in Winter (영동지역 겨울철 스캔라이다로 관측된 강수 이전 운저 인근 수상체의 광학 특성 분석)

  • Nam, Hyoung-Gu;Kim, Yoo-Jun;Kim, Seon-Jeong;Lee, Jin-Hwa;Kim, Geon-Tea;An, Bo-Yeong;Shim, Jae-Kwan;Jeon, Gye-hak;Choi, Byoung-Choel;Kim, Byung-Gon
    • Korean Journal of Remote Sensing
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    • v.34 no.2_1
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    • pp.237-248
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    • 2018
  • The vertical distribution of hydrometeor before precipitation near the cloud base has been analyzed using a scanning lidar, rawinsonde data, and Cloud-Resolving Storm Simulator (CReSS). This study mostly focuses on 13 Desember 2016 only. The typical synoptic pattern of lake-effect snowstorm induced easterly in the Yeongdong region. Clouds generated due to high temperature difference between 850 hPa and sea surface (SST) penentrated in the Yeongdong region along with northerly and northeasterly, which eventually resulted precipitation. The cloud base height before the precipitation changed from 750 m to 1,280 m, which was in agreement with that from ceilometer at Sokcho. However, ceilometer tended to detect the cloud base 50 m ~ 100 m below strong signal of lidar backscattering coefficient. As a result, the depolarization ratio increased vertically while the backscattering coefficient decreased about 1,010 m~1,200 m above the ground. Lidar signal might be interpreted to be attenuated with the penetration depth of the cloud layer with of nonspherical hydrometeor (snow, ice cloud). An increase in backscattering signal and a decrease in depolarization ratio occured in the layer of 800 to 1,010 m, probably being associated with an increase in non-spherical particles. There seemed to be a shallow liquid layer with a low depolarization ratio (<0.1) in the layer of 850~900 m. As the altitude increases in the 680 m~850 m, the backscattering coefficient and depolarization ratio increase at the same time. In this range of height, the maximum value (0.6) is displayed. Such a result can be inferred that the nonspherical hydrometeor are distributed by a low density. At this time, the depolarization ratio and the backscattering coefficient did not increase under observed melting layer of 680 m. The lidar has a disadvantage that it is difficult for its beam to penetrate deep into clouds due to attenuation problem. However it is promising to distinguish hydrometeor morphology by utilizing the depolarization ratio and the backscattering coefficient, since its vertical high resolution (2.5 m) enable us to analyze detailed cloud microphysics. It would contribute to understanding cloud microphysics of cold clouds and snowfall when remote sensings including lidar, radar, and in-situ measurements could be timely utilized altogether.

Susceptibility Loci Associations with Prostate Cancer Risk in Northern Chinese Men

  • Wang, Na-Na;Xu, Yong;Yang, Kuo;Wei, Dong;Zhang, Yao-Guang;Liu, Ming;Shi, Xiao-Hong;Liang, Si-Ying;Sun, Liang;Zhu, Xiao-Quan;Yang, Yi-Ge;Tang, Lei;Zhao, Cheng-Xiao;Wang, Xin;Chen, Xin;Hui, Juan;Zhang, Yu-Hong;Zhu, Ling;Yang, Fan;Zhang, Yu-Rong;Yang, Ze;Wang, Jian-Ye
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3075-3078
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    • 2013
  • Background: KLK3 gene products, like human prostate-specific antigen (PSA), are important biomarkers in the clinical diagnosis of prostate cancer (PCa). G protein-coupled receptor RFX6, C2orf43 and FOXP4 signaling plays important roles in the development of PCa. However, associations of these genes with PCa in northern Chinese men remain to be detailed. This study aimed to investigate their impact on occurrence and level of malignancy. Methods: All subjects were from Beijing and Tianjin, including 266 cases with prostate cancer and 288 normal individuals as controls. We evaluated associations between clinical covariates (age at diagnosis, prostate specific antigen, Gleason score, tumor stage and aggressive) and 6 candidate PCa risk loci, genotyped by PCR- high resolution melting curve and sequencing methods. Results: Case-control analysis of allelic frequency of PCa associated with PCa showed that one of the 6 candidate risk loci, rs339331 in the RFX6 gene, was associated with reduced risk of prostate cancer (odds ratio (OR) = 0.73, 95% confidence interval (CI) =0.57-0.94, P = 0.013) in northern Chinese men. In addition, subjects with CX (CC+TC) genotypes had a decreased risk for prostrate cancer compared to those carrying the TT homozygote (OR =0.64, 95% CI = 0.45- 0.90, P = 0.008). The TT genotype of 13q22 (rs9600079, T) was associated with tumor stage (P=0.044, OR=2.34, 95% CI=0.94-5.87). Other SNPs were not significantly associated with clinical covariates in prostate cancer (P > 0.05). Conclusions. rs339331 in the RFX6 gene may be associated with prostate cancer as a susceptibility locus in northern Chinese men.