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Detection of single-nucleotide polymorphism in RPB2 of Wolfiporia hoelen strains and assessment of its applicability for strain breeding (복령 균주의 RPB2 유전자 내 단일염기다형성 및 육종 활용성 분석)

  • Su Yeon, Kim;Mi-Jeong, Park;Seong Hwan, Kim;Kang-Hyeon, Ka
    • Journal of Mushroom
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    • v.20 no.4
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    • pp.199-207
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    • 2022
  • The demand for novel strains has been rising in the domestic market to increase the production of sclerotia from Wolfiporia hoelen. To improve strain breeding efficiency, we investigated whether single-nucleotide polymorphisms (SNPs) in the RNA polymerase II subunit (RPB2) gene, which may be linked to the mating type locus, are useful for distinguishing monokaryons from dikaryons in Korean W. hoelen strains. We designed a specific primer set to efficiently amplify a region of RPB2 using PCR with the genomic DNA of 12 cultivated strains and 31 wild strains of W. hoelen collected from Korea. Nucleotide sequences of the PCR-amplified RPB2 genes were determined and analyzed for the presence of SNPs among the 43 W. hoelen strains. Previously reported SNP loci were detected in the RPB2 gene of all W. hoelen strains tested. However, these previously reported SNP loci could not be applied to differentiate monokaryons from dikaryons in approximately one-third of Korean wild strains with homozygous genotypes. Three additional SNPs in the RPB2 gene, which may improve the ability to distinguish monokaryons from dikaryons, were identified by searching through the multiple sequence alignments of the 43 W. hoelen strains. The applicability of these three novel SNPs, together with the previously known SNPs, in the RPB2 gene to W. hoelen strain breeding was verified by examining the hybrid strains and their parental strains.

Phylogenetic and pathogenic traits of YHV3 and IHHNV detected from imported frozen shrimp (수입 냉동새우에서 검출된 YHV3와 IHHNV의 계통학 및 병원성 분석)

  • Baek, Eun Jin;Joeng, Ye Jin;Jeong, Min A;Park, Ji Yeon;Kim, Kwang Il
    • Journal of fish pathology
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    • v.35 no.1
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    • pp.27-40
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    • 2022
  • Yellow head virus (YHV), Infectious hypodermal and hematopoietic necrosis (IHHNV), Taura syndrome virus (TSV), and Infectious myositis virus (IMNV) cause serious mortality to Penaeidae shrimp in the aquaculture. In this study, YHV, IHHNV, TSV, and IMNV were surveyed from imported frozen shrimps between 2019 and 2020 via molecular diagnostic assay. Among 10 shrimp groups, YHV (n=1) and IHHNV (n=4) were detected by RT-PCR and PCR, respectively. From the phylogenetic analysis based on the partial ORF 1b region of YHV, YHV was classified into YHV genotype 3 (YHV3). And IHHNVs (n=2) detected from Litopenaeus vannamei belong to infectious IHHNV type 2. Although IHHNVs (n=2) identified from Penaeus monodon showed PCR positive results (MG 831F/R primer set), the sequences of ORF 2 and 3 were not amplified, suggesting that those samples might possess type A IHHNV related sequence of P. monodon. Furthermore, in the challenge test, even though PCR-detected isolates (YHV3/type A IHHNV related sequence or infectious IHHNV type 2) were not induced mortality to L. vannamei, viral genes were amplified suggesting that the viruses in the frozen shrimp could be non-pathogenic particles which are not enough to induce mortality.

Identification and Characterization of Polymorphic Microsatellite DNA Markers Using Next-generation Sequencing in Parapristipoma trilineatum (차세대 염기서열 분석법을 사용한 벤자리(Parapristipoma trilineatum)의 microsatellite 마커의 개발 및 유전학적 특성 분석)

  • Chun Mae Dong;Mi-Nan Lee;Jae Koo Noh;Jin Woo Park;Young-Ok Kim;Eun-Mi Kim
    • Journal of Life Science
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    • v.33 no.8
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    • pp.623-631
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    • 2023
  • This study was conducted to develop microsatellite markers in Parapristipoma trilineatum using next-generation sequencing. A total of 402,244,934 reads were generated on the Illumina Hiseq X Ten System, yielding 60,738,985,034 bp of sequences. The de novo assembly resulted in 1,320,995 contigs. A total of 952,326 contigs (0.016%) including 151 microsatellite loci were derived from the 1,320,995 contigs longer than 640 bp. A total of 34 primer sets were designed from the 151 microsatellite loci. As a result, 15 microsatellite loci were chosen and used for assuming population genetic parameters in the wild and farmed populations. The mean number of effective alleles was 12, ranging from 6 to 25. The observed heterozygosity (HO) and the expected heterozygosity (HE) ranged between 0.530 and 0.873, with an average of 0.750, and from 0.647 to 0.895, with an average of 0.793, respectively. According to these results, the developed set of 15 microsatellite markers is expected to be useful for the analysis of genetic characteristics in the population of P. trilineatum in Korea. There are requirements now for further genetic information, fishery resource management, breeding guidelines, support with the selection of breeds and studies on the effects of release, all of which will improve species conservation, and through future research, we aim to offer genetic foundational data with that goal.

Automated Detection and Segmentation of Bone Metastases on Spine MRI Using U-Net: A Multicenter Study

  • Dong Hyun Kim;Jiwoon Seo;Ji Hyun Lee;Eun-Tae Jeon;DongYoung Jeong;Hee Dong Chae;Eugene Lee;Ji Hee Kang;Yoon-Hee Choi;Hyo Jin Kim;Jee Won Chai
    • Korean Journal of Radiology
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    • v.25 no.4
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    • pp.363-373
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    • 2024
  • Objective: To develop and evaluate a deep learning model for automated segmentation and detection of bone metastasis on spinal MRI. Materials and Methods: We included whole spine MRI scans of adult patients with bone metastasis: 662 MRI series from 302 patients (63.5 ± 11.5 years; male:female, 151:151) from three study centers obtained between January 2015 and August 2021 for training and internal testing (random split into 536 and 126 series, respectively) and 49 MRI series from 20 patients (65.9 ± 11.5 years; male:female, 11:9) from another center obtained between January 2018 and August 2020 for external testing. Three sagittal MRI sequences, including non-contrast T1-weighted image (T1), contrast-enhanced T1-weighted Dixon fat-only image (FO), and contrast-enhanced fat-suppressed T1-weighted image (CE), were used. Seven models trained using the 2D and 3D U-Nets were developed with different combinations (T1, FO, CE, T1 + FO, T1 + CE, FO + CE, and T1 + FO + CE). The segmentation performance was evaluated using Dice coefficient, pixel-wise recall, and pixel-wise precision. The detection performance was analyzed using per-lesion sensitivity and a free-response receiver operating characteristic curve. The performance of the model was compared with that of five radiologists using the external test set. Results: The 2D U-Net T1 + CE model exhibited superior segmentation performance in the external test compared to the other models, with a Dice coefficient of 0.699 and pixel-wise recall of 0.653. The T1 + CE model achieved per-lesion sensitivities of 0.828 (497/600) and 0.857 (150/175) for metastases in the internal and external tests, respectively. The radiologists demonstrated a mean per-lesion sensitivity of 0.746 and a mean per-lesion positive predictive value of 0.701 in the external test. Conclusion: The deep learning models proposed for automated segmentation and detection of bone metastases on spinal MRI demonstrated high diagnostic performance.

Diagnostic Performance of Simulated Abbreviated MRI for Early-Stage Hepatocellular Carcinoma Screening: A Comparison to Conventional Dynamic Contrast-Enhanced MRI (초기 간암 선별 검사로서 단축 자기공명영상 검사의 진단능: 고식적 역동학적 자기공명영상검사와의 비교)

  • Eun Sol Lim;Sung Mo Kim;Sang Soo Shin;Suk Hee Heo;Jong Eun Lee;Yong Yeon Jeong
    • Journal of the Korean Society of Radiology
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    • v.82 no.5
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    • pp.1218-1230
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    • 2021
  • Purpose To compare the per-patient diagnostic performance of simulated abbreviated MRI (AMRI) to that of conventional MRI (CMRI) with full-sequence dynamic gadoxetic acid (GA) enhancement for early-stage hepatocellular carcinoma (HCC) screening in high-risk patients. Materials and Methods A total of 201 consecutive patients at high-risk for HCC, who underwent 3T liver MRI, were included in this retrospective study. The AMRI protocol comprised T2-weighted imaging, hepatobiliary phase imaging after GA injection, and diffusion-weighted imaging. For each patient, two AMRI and CMRI image sets were independently reviewed by two radiologists. Inter-reader agreement was assessed using Cohen's kappa value. A composite reference standard was used to determine the diagnostic performance of each image set for each reader. Results A total of 93 HCCs were detected in 79 patients. The inter-reader agreement was almost perfect for both image sets (κ = 0.839, 0.948). In AMRI, the per-patient sensitivity and negative predictive values (NPV) were 94.9% and 96.4%, respectively. In CMRI, the per-patient sensitivity and NPV were 96.2% and 97.5%, respectively. Conclusion AMRI, using only three sequences, had a comparable diagnostic performance to CMRI in screening early-stage HCC. AMRI could be an alternative HCC screening tool for high-risk HCC patients.

A Set of Anthocyanin Biosynthetic Genes are Differentially Expressed in Strawberry (Fragaria x ananassa cv Maehyang) during the Fruit Development Process (매향 딸기로부터 anthocyanin 합성 유전자의 분리 및 과실발달 과정에서의 발현 분석)

  • Bae, Ki-Suk;Kih, Joon-Yeong;Pyee, Jae-Ho
    • Journal of Life Science
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    • v.18 no.2
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    • pp.234-240
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    • 2008
  • Anthocyanin synthesis in strawberry (Fragaria x ananassa cv Maehyang) begins approximately 26 days postflowering and continued throughout fruit ripening. A set of cDNA clones encoding the anthocyanin biosynthetic enzymes were isolated from strawberry. A pair of primers were designed for polymerase chain reaction (PCR) through the comparison of the nucleotide sequences of homologous genes from diverse plants. Reverse transcriptase-PCRs were performed using cDNA synthesized from ripe fruit total RNA and the primers corresponding to each gene. Eight genes of the anthocyanin pathway were cloned and confirmed by sequencing to code for phenylalanine ammonia lyase (PAL), 4-cummarate CoA ligase (4CL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone-3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), anthocyanidine synthase (ANS), UDP-glucose:flavonoid-3-O-glucosyl-transferase (UFGT). Northern analyses showed that the corresponding genes were differentially expressed during the fruit development process. All genes except PAL were predominantly expressed in fruit. Expression of PAL, DFR and ANS was detected 10 days postflowering at the early stage of fruit development, declined for a while and sharply increased 22 days postflowering then showed a peak 34 days postflowering. The other genes, however, were not expressed up to 22 or 30 days postflowering when the initial fruit ripening events occur at the time of initiation of anthocyanin accumulation. The onset of anthocyanin synthesis in ripening strawberry coincides with a coordinated induction of the anthocyanin pathway genes, suggesting the involvement of regulatory genes. We propose that at least two different regulatory mechanisms playa role in the biosynthesis of anthocyanin during color development of strawberry.

Analysis of shopping website visit types and shopping pattern (쇼핑 웹사이트 탐색 유형과 방문 패턴 분석)

  • Choi, Kyungbin;Nam, Kihwan
    • Journal of Intelligence and Information Systems
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    • v.25 no.1
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    • pp.85-107
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    • 2019
  • Online consumers browse products belonging to a particular product line or brand for purchase, or simply leave a wide range of navigation without making purchase. The research on the behavior and purchase of online consumers has been steadily progressed, and related services and applications based on behavior data of consumers have been developed in practice. In recent years, customization strategies and recommendation systems of consumers have been utilized due to the development of big data technology, and attempts are being made to optimize users' shopping experience. However, even in such an attempt, it is very unlikely that online consumers will actually be able to visit the website and switch to the purchase stage. This is because online consumers do not just visit the website to purchase products but use and browse the websites differently according to their shopping motives and purposes. Therefore, it is important to analyze various types of visits as well as visits to purchase, which is important for understanding the behaviors of online consumers. In this study, we explored the clustering analysis of session based on click stream data of e-commerce company in order to explain diversity and complexity of search behavior of online consumers and typified search behavior. For the analysis, we converted data points of more than 8 million pages units into visit units' sessions, resulting in a total of over 500,000 website visit sessions. For each visit session, 12 characteristics such as page view, duration, search diversity, and page type concentration were extracted for clustering analysis. Considering the size of the data set, we performed the analysis using the Mini-Batch K-means algorithm, which has advantages in terms of learning speed and efficiency while maintaining the clustering performance similar to that of the clustering algorithm K-means. The most optimized number of clusters was derived from four, and the differences in session unit characteristics and purchasing rates were identified for each cluster. The online consumer visits the website several times and learns about the product and decides the purchase. In order to analyze the purchasing process over several visits of the online consumer, we constructed the visiting sequence data of the consumer based on the navigation patterns in the web site derived clustering analysis. The visit sequence data includes a series of visiting sequences until one purchase is made, and the items constituting one sequence become cluster labels derived from the foregoing. We have separately established a sequence data for consumers who have made purchases and data on visits for consumers who have only explored products without making purchases during the same period of time. And then sequential pattern mining was applied to extract frequent patterns from each sequence data. The minimum support is set to 10%, and frequent patterns consist of a sequence of cluster labels. While there are common derived patterns in both sequence data, there are also frequent patterns derived only from one side of sequence data. We found that the consumers who made purchases through the comparative analysis of the extracted frequent patterns showed the visiting pattern to decide to purchase the product repeatedly while searching for the specific product. The implication of this study is that we analyze the search type of online consumers by using large - scale click stream data and analyze the patterns of them to explain the behavior of purchasing process with data-driven point. Most studies that typology of online consumers have focused on the characteristics of the type and what factors are key in distinguishing that type. In this study, we carried out an analysis to type the behavior of online consumers, and further analyzed what order the types could be organized into one another and become a series of search patterns. In addition, online retailers will be able to try to improve their purchasing conversion through marketing strategies and recommendations for various types of visit and will be able to evaluate the effect of the strategy through changes in consumers' visit patterns.

The Brassica rapa Tissue-specific EST Database (배추의 조직 특이적 발현유전자 데이터베이스)

  • Yu, Hee-Ju;Park, Sin-Gi;Oh, Mi-Jin;Hwang, Hyun-Ju;Kim, Nam-Shin;Chung, Hee;Sohn, Seong-Han;Park, Beom-Seok;Mun, Jeong-Hwan
    • Horticultural Science & Technology
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    • v.29 no.6
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    • pp.633-640
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    • 2011
  • Brassica rapa is an A genome model species for Brassica crop genetics, genomics, and breeding. With the completion of sequencing the B. rapa genome, functional analysis of the genome is forthcoming issue. The expressed sequence tags are fundamental resources supporting annotation and functional analysis of the genome including identification of tissue-specific genes and promoters. As of July 2011, 147,217 ESTs from 39 cDNA libraries of B. rapa are reported in the public database. However, little information can be retrieved from the sequences due to lack of organized databases. To leverage the sequence information and to maximize the use of publicly-available EST collections, the Brassica rapa tissue-specific EST database (BrTED) is developed. BrTED includes sequence information of 23,962 unigenes assembled by StackPack program. The unigene set is used as a query unit for various analyses such as BLAST against TAIR gene model, functional annotation using MIPS and UniProt, gene ontology analysis, and prediction of tissue-specific unigene sets based on statistics test. The database is composed of two main units, EST sequence processing and information retrieving unit and tissue-specific expression profile analysis unit. Information and data in both units are tightly inter-connected to each other using a web based browsing system. RT-PCR evaluation of 29 selected unigene sets successfully amplified amplicons from the target tissues of B. rapa. BrTED provided here allows the user to identify and analyze the expression of genes of interest and aid efforts to interpret the B. rapa genome through functional genomics. In addition, it can be used as a public resource in providing reference information to study the genus Brassica and other closely related crop crucifer plants.

Phylogentic Position, Pigment Content and Optimal Growth Condition of the Unicellular Hydrogen-Producing Cyanobacterial Strains from Korean Coasts (한국 연안산 단세포성 수소생산 남세균 종주들의 분류계통, 색소함량 및 최적성장 환경)

  • PARK, JONG-WOO;KIM, JU HEE;CHO, AE-RA;JUNG, YUN-DUK;KIM, PYOUNG JOONG;KIM, HYUNG-SEOP;YIH, WONHO
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.20 no.3
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    • pp.131-140
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    • 2015
  • To set up unicellular cyanobacterial strains with photo-biological $H_2$ production potential, live samples were repeatedly collected from 68 stations in the coastal zone of Korea for the four years since 2005. Among 77 cyanobacterial strains established six (KNU strains, CB-MAL002, 026, 031, 054, 055 and 058) were finally chosen as the excellent strains for $H_2$ production with $H_2$ accumulation over 0.15 mL $H_2\;mL^{-1}$ under general basic $H_2$ production conditions as well as positive $H_2$ production for more than 60 hr. To explore optimum procedures for higher $H_2$ production efficiency of the six cyanobacterial strains, the inter-strain differences in the growth rate under the gradients of water temperature and salinity were investigated. The maximum daily growth rates of the six strains ranged from 1.78 to 2.08, and all of them exhibited $N_2-fixation$ ability. Based on the similarity of the 16S rRNA sequences, all the test strains were quite close to Cyanothece sp. ATCC51142 (99%). The six strains, however, were grouped into separate clades from strain ATCC51142 in the molecular phylogeny diagram. Chlorophyll- a content was 3.4~7.8% of the total dried weight, and the phycoerythrin and phycocyanin contents were half of those in the Atlantic strain, Synechococcus sp. Miami BG03511. The growth of the six strains was significantly suppressed at temperatures above the optimal range, $30{\sim}35^{\circ}C$, to be nearly stopped at $40^{\circ}C$. The growth was not inhibited by high salinities of 30 psu salinity in all the strains while strain CB055 maintained its high growth rate at low salinities down to 15 psu. The euryhaline strains like CB055 might support massive biotechnological cultivation systems using natural basal seawater in temperate latitudes. base seawater. The biological and ecophysiological characteristics of the test strains may contribute to designing the optimal procedures for photo-biological $H_2$ production by unicellular cyanobacteria.

Use of Human Serum Albumin Fusion Tags for Recombinant Protein Secretory Expression in the Methylotrophic Yeast Hansenula polymorpha (메탄올 자화효모 Hansenula polymorpha에서의 재조합 단백질 분비발현을 위한 인체 혈청 알부민 융합단편의 활용)

  • Song, Ji-Hye;Hwang, Dong Hyeon;Oh, Doo-Byoung;Rhee, Sang Ki;Kwon, Ohsuk
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.17-25
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    • 2013
  • The thermotolerant methylotrophic yeast Hansenula polymorpha is an attractive model organism for various fundamental studies, such as the genetic control of enzymes involved in methanol metabolism, peroxisome biogenesis, nitrate assimilation, and resistance to heavy metals and oxidative stresses. In addition, H. polymorpha has been highlighted as a promising recombinant protein expression host, especially due to the availability of strong and tightly regulatable promoters. In this study, we investigated the possibility of employing human serum albumin (HSA) as the fusion tag for the secretory expression of heterologous proteins in H. polymorpha. A set of four expression cassettes, which contained the methanol oxidase (MOX) promoter, translational HSA fusion tag, and the terminator of MOX, were constructed. The expression cassettes were also designed to contain sequences for accessory elements including His8-tag, $2{\times}(Gly_4Ser_1)$ linkers, tobacco etch virus protease recognition sites (Tev), multi-cloning sites, and strep-tags. To determine the effects of the size of the HSA fusion tag on the secretory expression of the target protein, each cassette contained the HSA gene fragment truncated at a specific position based on its domain structure. By using the Green fluorescence protein gene as the reporter, the properties of each expression cassette were compared in various conditions. Our results suggest that the translational HSA fusion tag is an efficient tool for the secretory expression of recombinant proteins in H. polymorpha.