• Title/Summary/Keyword: MOLECULAR MARKER

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Serum Talin-1 is a Potential Novel Biomarker for Diagnosis of Hepatocellular Carcinoma in Egyptian Patients

  • Youns, Mahmoud M.;Abdel Wahab, Abdel Hady A.;Hassan, Zeinab A.;Attia, Mohamed S.
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3819-3823
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    • 2013
  • Background: Hepatocellular carcinoma (HCC) is a major cause of cancer mortality worldwide. The outcome of HCC depends mainly on its early diagnosis. To date, the performance of traditional biomarkers is unsatisfactory. Talins were firstly identified as cytoplasmic protein partners of integrins but Talin-1 appears to play a crucial role in cancer formation and progression. Our study was conducted to assess the diagnostic value of serum Talin-1 (TLN1) compared to the most feasible traditional biomarker alpha-fetoprotein (AFP) for the diagnosis of HCC. Methods: TLN1 was detected using enzyme linked immunosorbent assay (ELISA) in serum samples from 120 Egyptian subjects including 40 with HCC, 40 with liver cirrhosis (LC) and 40 healthy controls (HC). Results: ROC curve analysis was used to create a predictive model for TLN1 relative to AFP in HCC diagnosis. Serum levels of TLN1 in hepatocellular carcinoma patients were significantly higher compared to the other groups (p<0.0001). The diagnostic accuracy of TLN1 was higher than that of AFP regarding sensitivity, specificity, positive predictive value and negative predictive value in diagnosis of HCC. Conclusions: The present study showed for the first time that Talin-1 (TLN1) is a potential diagnostic marker for HCC, with a higher sensitivity and specificity compared to the traditional biomarker AFP.

Analysis of RNA Polymerase Beta Subunit (rpoB) Gene Sequences for the Discrimination of Cyanobacteria Anabaena Species (남조세균 Anabaena 종 구분을 위한 RNA Polymerase Beta Subunit (rpoB) 유전자 염기서열 분석)

  • Cheon, Ju-Yong;Lee, Min-Ah;Ki, Jang-Seu
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.268-274
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    • 2011
  • Anabaena (Cyanobacteria, Nostocales) are important for water quality controls, because they are often responsible for freshwater green tides; moreover, some species are reported to produce hepatotoxin. In this study, we sequenced RNA polymerase beta subunit (rpoB) gene of Anabaena, and evaluated their sequences for the potential use of a molecular taxonomic marker in this taxon. Anabaena rpoB showed low DNA similarity and high genetic divergences when compared those of 16S rRNA, and the molecular differences were statistically significant (Student t-test, p<0.01). Parsimony analyses showed the rpoB gene evolves 4.8-fold faster than 16S rRNA. In addition, phylogeny of the rpoB gene separated each Anabaena strain more clearly compared with a 16S rRNA tree. These results suggest that the rpoB gene is a useful marker for the molecular phylogenetics and the species discrimination of Anabaena.

Use of Anti-Phosphohistone H3 (PHH3) as a Mitosis Marker for Classifying Pulmonary Carcinoid Tumors

  • Seo, Bo-Ram;Hong, Young-Seob;Choi, Phil-Jo;Um, Soo-Jung;Seo, Jeong-Wook;Roh, Mee-Sook
    • Biomedical Science Letters
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    • v.17 no.3
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    • pp.197-202
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    • 2011
  • Mitosis count is one of the most helpful morphologic features for distinguishing pulmonary typical carcinoid (TC) from atypical carcinoid (AC). However, identifying areas of highest mitotic activity is tedious and time-consuming, and mitosis count may vary substantially among pathologists. Anti-phosphohistone H3 (PHH3) is an antibody that specifically detects histone H3 only when phosphorylated at serine 10 or serine 28, an event that is concurrent with mitotic chromatin condensation and not observed during apoptosis. In this study, immunohistochemical staining for PHH3 was performed to determine whether PHH3 was a reliable and objective mitosis-specific marker for pulmonary carcinoid tumors. Seventeen cases of surgically resected pulmonary carcinoid tumors (12 TCs and 5 ACs) were obtained and classified according to the 2004 World Health Organization classification. Mitotic counts determined by PHH3 correlated to ones determined by hematoxylin and eosin (H&E) staining; however, PHH3 mitotic counts (mean mitotic counts: 1 in TCs and 3.2 in ACs) were slightly higher than H&E mitotic counts (mean mitotic counts: 0.25 in TCs and 1.8 in ACs). The mitotic counts determined by experienced observer were more correlated to those determined by inexperienced observer with the PHH3-based method (R=0.968, P<0.001) rather than H&E staining (R=0.658, P<0.001). These results suggest that the PHH3 mitotic counting method was more sensitive and simple for detecting mitoses compared to traditional H&E staining. Therefore, PHH3 immunohistochemistry may contribute to more accurate and reproducible diagnosis of pulmonary carcinoid tumors and may be a valuable aid for administrating appropriate clinical treatment.

Development of PCR-Based Molecular Marker for Detection of Xanthomonas campestris pv. campestris Race 6, the Causative Agent of Black Rot of Brassicas

  • Afrin, Khandker Shazia;Rahim, Md Abdur;Rubel, Mehede Hassan;Park, Jong-In;Jung, Hee-Jeong;Kim, Hoy-Taek;Nou, Ill-Sup
    • The Plant Pathology Journal
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    • v.36 no.5
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    • pp.418-427
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    • 2020
  • Xanthomonas campestris pv. campestris (Xcc), the pathogen of black rot which is the most destructive disease of Brassica vegetables throughout the world. Here, we reported two novel sequence-characterized amplified region (SCAR) markers (i.e., XccR6-60 and XccR6-67) for the detection of Xcc race 6 via re-alignment of the complete genome sequences of Xcc races/strains/pathovars. The specificity of SCAR primer sets was verified by mean of PCR amplification using the genomic DNA template of Xcc races/strains/pathovars and two other plant infecting bacterial strains. The PCR result revealed that the XccR6-60 and XccR6-67 primer sets amplified 692-bp and 917-bp DNA fragments, respectively, specifically from race 6, while no visible amplification was detected in other samples. In addition, the SCAR primers were highly sensitive and can detect from a very low concentration of genomic DNA of Xcc race 6. However, the complete genome sequence of Xcc race 6 is not yet publicly available. Therefore, the cloning and sequencing of XccR6-60 and XccR6-67 fragments from race 6 provide more evidence of the specificity of these markers. These results indicated that the newly developed SCAR markers can successfully, effectively and rapidly detect Xcc race 6 from other Xcc races/strains/pathovars as well as other plant pathogenic bacteria. This is the first report for race-specific molecular markers for Xcc race 6.

Comparison of Plant-derived Carbonaceous Components (Organic Molecular Markers and 14carbon) in PM2.5 in Summer and Autumn at Kazo, Japan

  • Sasaka, Kouki;Wang, Qingyue;Sakamoto, Kazuhiko
    • Asian Journal of Atmospheric Environment
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    • v.11 no.3
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    • pp.165-175
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    • 2017
  • In Japan, the primary carbonaceous particles emitted from motor vehicles and waste incinerators have been reduced due to strict regulations against exhaust gas. However, the relative contribution of carbonaceous particles derived from plants and biomass has been increasing. Accordingly, compositional analysis of carbonaceous particles has become increasingly important to determine the sources and types of particles produced. To reveal the sources of the organic particles contained in particulate matter with diameters of ${\leq}2.5{\mu}m$ ($PM_{2.5}$) and the processes involved in their generation, we analyzed molecular marker compounds (2-methyltetrols, cis-pinonic acid, and levoglucosan) derived from the plants and biomass in the $PM_{2.5}$ collected during daytime- and nighttime-sampling periods in summer (July and August) and autumn (November) in Kazo, which is in the northern area of Saitama prefecture, Japan. We also measured $^{14}C$ carbonaceous concentrations in the same $PM_{2.5}$ samples. The concentrations of 2-methyltetrols were higher in the summer than in the autumn. Because the deciduous period overlaps with this decrease in the levels of 2-methyltetrols, we considered the emission source to broad-leaved trees. In contrast, the emission source of the cis-pinonic acid precursor was considered to be conifers, because its concentration remained almost constant throughout the year. The concentration of levoglucosan was considerably increased in the autumn due to frequent biomass open burning. The ratio of plant-derived carbon to total carbon, obtained by measuring of $^{14}C$, in summer $PM_{2.5}$ sample was higher in the nighttime, and could be influenced by anthropogenic sources during the daytime.

Source Profile of Road Dust for Statistical Apportionment Modeling in Seoul (통계 수용모델을 위한 서울시 도로변 화학성분 원인 프로파일)

  • Park, Da-Jeong;Han, Young-Ji;Lee, Ji-Yi;Lee, Kwang-Yul;Cho, In-Hwan;Park, Eun Ha;Yi, Seung-Muk;Bae, Min-Suk
    • Journal of Korean Society for Atmospheric Environment
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    • v.31 no.2
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    • pp.105-117
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    • 2015
  • Sources related to road dust is one of the biggest sources, which is responsible for a large portion of emission. In particular, PM2.5 is a potential cause for respiratory diseases, thus it should be managed and a mitigation plan using results of statistical apportionment modeling such as chemical mass balance needs to be established. Recently, identifying sources of PM2.5 and analyzing the contribution of the road dust through a contribution assessment is required. Therefore, this study provides the chemical source profiles of PM2.5 using IC, GC/MS, OCEC, and XRF for both paved sidewalk and paved roadway collected at seven different sampling sites. As a result, for paved sidewalk, $NH{_4}^+$ (70%), $NO{_3}^-$ (12%), $PO{_4}^-$ (9%), and $SO{_4}^{2-}$ (9%) have been analyzed in PM2.5 mass. Major molecular marker such as Si has been indicated as $12.0{\pm}3.4%$ and $13.6{\pm}6.9%$ for paved sidewalk and roadway, respectively. PAHs such as Fluoranthene, Pyrene, Chrysene, and 1,3,5-Triphenylbenzene are suggested as molecular markers for road dust.

Identification of Species and Sex of Korean Roe Deer (Capreolus pygargus tianschanicus) Using SRY and CYTB Genes

  • Han, Sang-Hyun;Cho, In-Cheol;Lee, Sung-Soo;Tandang, Leoncia;Lee, Hang;Oh, Hong-Shik;Kim, Byoung-Soo;Oh, Moon-You
    • Animal cells and systems
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    • v.11 no.2
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    • pp.165-168
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    • 2007
  • The nucleotide sequences of a male-specific marker sex determining region Y (SRY) gene and a mitochondrial cytochrome B (CYTB) gene were characterized and analyzed to establish a molecular method for identification of species and sex of Korean roe deer (Capreolus pygargus tianschanicus). Similarity search result of SRY sequences showed very similar result to those reported in Moose (Alces alces) and Reindeer (Rangifer tarandus), both of which had 95.9% similarity in identity. CYTB genes were very similar to those reported in Siberian roe deer (C. pygargus pygargus) which had 98.6% similarity and not to European roe deer (C. capreolus), suggesting that the DNA samples tested were of Siberian roe deer lineage. Polymerase chain reaction (PCR)-based sex typing successfully discriminated between carcasses of male and female roe deer. Males had SRY band on agarose gels and females did not. The result of this molecular sex typing provided similar information with that obtained by genital organ observation. Therefore, this molecular method using male specific marker SRY and mitochondrial CYTB genes would be very useful for identification of the species and sex of the carcass remains of roe deer.

Development of Molecular Markers for Xanthomonas axonopodis Resistance in Soybean

  • Kim Ki-Seung;Van Kyujung;Kim Moon Young;Lee Suk-Ha
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.5
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    • pp.429-433
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    • 2004
  • A single recessive gene, rxp, controls the bacterial leaf pustule (BLP) resistance in soybean and in our previous article, it has been mapped on linkage group (LG) D2 of molecular genetic map of soybean. A total of 130 recombinant inbred lines (RILs) from a cross between BLP-resistant SS2-2 and BLP-susceptible Jangyeobkong were used to identify molecular markers linked to rxp. Fifteen simple sequence repeat (SSR) markers on LG D2 were screened to construct a genetic map of rxp locus. Only four SSR markers, Satt135, Satt372, Satt448, and Satt486, showed parental polymorphisms. Using these markers, genetic scaffold map was constructed covering 26.2cM. Based on the single analysis of variance, Satt372 among these four SSR markers was the most significantly associated with the resistance to BLP. To develop new amplified fragment length polymorphism (AFLP) marker linked to the resistance gene, bulked segregant analysis (BSA) was employed. Resistance and susceptible bulks were made by pooling equal amount of genomic DNAs from ten of each in the segregating population. A total of 192 primer combinations were used to identify specific bands to the resistance, selecting three putative AFLP markers. These AFLP markers produced the fragment present in SS2-2 and the resistant bulk, and not in Jangyeobkong and the susceptible bulk. Linkage analysis revealed that McctEact97 $(P=0.0004,\;R^2=14.67\%)$ was more significant than Satt372, previously reported as the most closely linked marker.

Identification of SNPs tightly linked to the QTL for pod shattering in soybean[Glycine max (L.) Merr.]

  • Kim, Kyung-Ryun;Kim, Kyung Hye;Go, Hong Min;Lee, Ju Seok;Moon, Jung-Kyung;Ha, Bo-Keun;Jeong, Soon-Chun;Kim, Namshin;Kang, Sungtaeg
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.146-146
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    • 2017
  • The pod shattering or dehiscence is essential for the propagation of pod-bearing plant species in the wild, but it causes significant yield losses during harvest of domesticated crop plants. Identifying novel molecular makers, which are linked to seed-shattering genes, is needed to employ the molecular marker-assisted selection for efficiently developing shattering-resistant soybean varieties. In this study, a genetic linkage map was constructed using 115 recombinant inbred lines (RILs) developed from crosses between the pod shattering susceptible variety, Keunol, and resistant variety, Sinpaldal. A 180 K Axiom(R) SoyaSNPs data and pod shattering data from two environments in 2001 and 2015 were used to identify quantitative trait loci (QTL) for pod shattering. A major QTL was identified between two flanking single nucleotide polymorphism (SNP) markers, AX-90320801 and AX-90306327 on chromosome 16 with 1.3 cM interval, 857 kb of physical range. In sequence, genotype distribution analysis was conducted using extreme phenotype RILs. This could narrow down the QTL down to 153 kb on the physical map and was designated as qPDH1-KS with 6 annotated gene models. All exons within qPDH1-KS were sequenced and the 6 polymorphic SNPs affecting the amino acid sequence were identified. To develop universally available molecular markers, 38 Korean soybean cultivars were investigated by the association study using the 6 identified SNPs. Only two SNPswere strongly associated with the pod shattering. These two identified SNPs will help to identify the pod shattering responsible gene and to develop pod shattering-resistant soybean plants using marker-assisted selection.

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Construction of a Bacterial Artificial Chromosome Library Containing Large BamHI Genomic Fragments from Medicago truncatula and Identification of Clones Linked to Hypernodulating Genes

  • Park So-Yeon;Nam Young-Woo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.256-263
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    • 2006
  • In the model legume Medicago truncatula, two mutants, sickle and sunn, exhibit morphologically and genetically distinct hypernodulation phenotypes. However, efforts to isolate the single recessive and single semidominant genes for sickle and sunn, respectively, by map-based cloning have so far been unsuccessful, partly due to the absence of clones that enable walks from linked marker positions. To help resolve these difficulties, a new bacterial artificial chromosome (BAC) library was constructed using BamHI-digested genomic fragments. A total of 23,808 clones were collected from ligation mixtures prepared with double-size-selected high-molecular-weight DNA. The average insert size was 116 kb based on an analysis of 88 randomly selected clones using NotI digestion and pulsed-field gel electrophoresis. About 18.5% of the library clones lacked inserts. The frequency of the BAC clones carrying chloroplast or mitochondrial DNA was 0.98% and 0.03%, respectively. The library represented approximately 4.9 haploid M. truncatula genomes. Hybridization of the BAC clone filters with a $C_{0}t-l$ DNA probe revealed that approximately 37% of the clones likely carried repetitive sequence-enriched DNA. An ordered array of pooled BAC DNA was screened by polymerase chain reactions using 13 sequence-characterized molecular markers that belonged to the eight linkage groups. Except for two markers, one to five positive BAC clones were obtained per marker. Accordingly, the sickle- and sunn-linked BAC clones identified herein will be useful for the isolation of these biotechnologically important genes. The new library will also provide clones that fill the gaps between preexisting BAC contigs, facilitating the physical mapping and genome sequencing of M. truncatula.