• Title/Summary/Keyword: Marker-set

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A fast and reliable polymerase chain reaction method based on short interspersed nuclear elements detection for the discrimination of buffalo, cattle, goat, and sheep species in dairy products

  • Cosenza, Gianfranco;Iannaccone, Marco;Gallo, Daniela;Pauciullo, Alfredo
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.891-895
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    • 2019
  • Objective: Aim of present study was the set up of a fast and reliable protocol using species-specific markers for the quali-quantitative analysis of DNA and the detection of ruminant biological components in dairy products. For this purpose, the promoter of the gene coding for the ${\alpha}$-lactoalbumin (LALBA) was chosen as possible candidate for the presence of short interspersed nuclear elements (SINEs). Methods: DNA was isolated from somatic cells of 120 individual milk samples of cattle (30), Mediterranean river buffalo (30), goat (30), and sheep (30) and the gene promoter region (about 600/700 bp) of LALBA (from about 600 bp upstream of exon 1) has been sequenced. For the development of a single polymerase chain reaction (PCR) protocol that allows the simultaneous identification of DNA from the four species of ruminants, the following internal primers pair were used: 5'-CACTGATCTTAAAGCTCAGGTT-3' (forward) and 5'-TCAGA GTAGGCCACAGAAG-3' (reverse). Results: Sequencing results of LALBA gene promoter region confirmed the presence of SINEs as monomorphic "within" and variable in size "among" the selected species. Amplicon lengths were 582 bp in cattle, 592 bp in buffalo, 655 in goat and 729 bp in sheep. PCR specificity was demonstrated by the detection of trace amounts of species-specific DNA from mixed sources ($0.25ng/{\mu}L$). Conclusion: We developed a rapid PCR protocol for the quali-quantitative analysis of DNA and the traceability of dairy products using a species-specific marker with only one pair of primers. Our results validate the proposed technique as a suitable tool for a simple and inexpensive (economic) detection of animal origin components in foodstuffs.

A literature review on diagnostic markers and subtype classification of children with speech sound disorders (원인을 모르는 말소리장애의 하위유형 분류 및 진단 표지에 관한 문헌 고찰)

  • Yi, Roo-Dah;Kim, Soo-Jin
    • Phonetics and Speech Sciences
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    • v.14 no.2
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    • pp.87-99
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    • 2022
  • A review regarding indicators used in Korean research is needed to develop a diagnostic marker system for Korean children with speech sound disorders (SSD). This literature review examined the research conducted to reveal the characteristics of children with SSD of unknown origin in Korea. The researchers in Korea used diverse variables as indicators to identify the natures of children with SSD. These included indicators related to external characteristics of speech sound and comorbid features other than external aspects of speech sound. The attention has been focused on specific indicators so far. This result implies that some indicators may still require closer study in various aspects due to their influence, and some may require more attention due to the limited number of research. This article argues that more research is necessary to comprehensively describe the unique characteristics of children with SSD of unknown origin and suggests a direction for future research regarding diagnostic markers and subtype classification of SSD. It also proposes potential diagnostic markers and a set of assessments for the subtype classification of SSD.

Effect of Defined KSOM Medium on the Development of 1-antitrypsin Transgenic Nuclear Transfer Bovine Embryos

  • M.M.U. Bhuiyan;J.K. Cho;G. Jang;Park, E.S.;S.K. Kang;Lee, B.C.;W.S. Hwang
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.74-74
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    • 2002
  • Production of u 1-antitrypsin ($\alpha$AT) in transgenic cows has a great value in the field of medicine. The present study was conducted to determine the effect of chemically defined KSOM media on in vitro development of bovine transgenic nuclear transfer (NT) embryos. An expression plasmid for human $\alpha$AT was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human $\alpha$AT target gene into a pcDNA3 plasmid. Cumulus cells as donor nuclei in NT were collected from a Holstein cow and transfected by lipid-mediated method using FuGene6 (Roche Molecular Biochemicals, USA) as reagent. GFP expressed cumulus cells were introduced into recipient oocytes under DIC microscopy equipped with FITC filter set. After electrical fusion and chemical activation, reconstructed embryos were cultured in 1) SOF + 0.8% BSA, 2) KSOM + 0.8% BSA, 3) KSOM + 10% FBS and 4) KSOM +0.01% PVA for 192 h at 39$^{\circ}C$ with 5% $CO_2$, 5% $O_2$ and 90% $N_2$in humidified condition. The development of the embryos was recorded and the GFP expression in blastocyst was determined under FITC filter. The average fusion rate was 73.8% (251/340; n=8). The development rates to 2-4 cells, morula, blastocysts and expression rates in blastocysts varied from 70.3 to 76.5%, 30.2 to 33.8%, 25.4 to 33.8% and 11.8 to 15.6%, respectively. The difference in development and expression rates of embryos among 4 culture groups was not significant (P>0.05). This study indicates that chemically defined KSOM medium is also able to support development of bovine transgenic NT embryos at similar rate of SOF or KSOM supplemented with BSA or serum.

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標記"了"的認知解析考察

  • Lee, Ju-Eun
    • 중국학논총
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    • no.53
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    • pp.1-16
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    • 2017
  • The grammatical meaning of "了", a marker of aspects in Mandarin has been described differently among researchers, three of which are to express the end, a certain phase of event and the start of an event or a state. This paper consists of four parts. Set out below is a summary of main points of each part. The first chapter elaborates on the results of previous related studies, points out the purpose and meaning of the current study. Chapter two introduces marked "了" including concepts of the meaning and "Done", "complete whole", "realization" Corresponds to the case study. And explains syntactic and semantic features of the "了", highlights the look "of the了" problems of syntax and semantics, "了2" made three domains Limit 3 Including of the knowledge. This chapter holds that its grammartical meaning can be described as follows an event has taken place or a state has formed prior to a certain reference time, and that the grammartical meanings mentioned above can be considered sub - grammartical meanings of "了",which result from different prominence of the phases of an event. Chapter three explains tag component of the "了1", "了2". It discusses limited constraints, first place for "了" syntactic meaning, "The End" and "extension" of the opposition for "了" is looming around the root system, "了" and "着" sentence structure and semantic and syntactic forms. Chapter four discusses bounded features of the 了. It mainly focuses on "了"of bounded / unbounded function. It analyses the functions of components and explains a bounded function of "了1" and a bounded function of "了2". Chapter five is the conclusion. It consolidates and summarises the previous contents of the paper and points out the conclusion.

Complete Chloroplast Genome assembly and Annotation of Milk Thistle (Silybum marianum) and Phylogenetic Analysis

  • Hwajin Jung;Yedomon Ange Bovys Zoclanclounon;Jeongwoo Lee;Taeho Lee;Jeonggu Kim;Guhwang Park;Keunpyo Lee;Kwanghoon An;Jeehyoung Shim;Joonghyoun Chin;Suyoung Hong
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.210-210
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    • 2022
  • Silybum marianum is an annual or biennial plant from the Asteraceae family. It can grow in low-nutrient soil and drought conditions, making it easy to cultivate. From the seed, a specialized plant metabolite called silymarin (flavonolignan complex) is produced and is known to alleviate the liver from hepatitis and toxins damages. To infer the phylogenetic placement of a Korean milk thistle, we conducted a chloroplast assembly and annotation following by a comparison with existing Chinese reference genome (NC_028027). The chloroplast genome structure was highly similar with an assembly size of 152,642 bp, an 153,202 bp for Korean and Chinese milk thistle respectively. Moreover, there were similarities at the gene level, coding sequence (n = 82), transfer RNA (n = 31) and ribosomal RNA (n = 4). From all coding sequences gene set, the phylogenetic tree inference placed the Korean cultivar into the milk thistle clade; corroborating the expected tree. Moreover, an investigation the tree based only on the ycf1 gene confirmed the same tree; suggesting that ycf1 gene is a potential marker for DNA barcoding and population diversity study in milk thistle genus. Overall, the provided data represents a valuable resource for population genomics and species-centered determination since several species have been reported in the Silybum genus.

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Implementation of Markerless Augmented Reality with Deformable Object Simulation (변형물체 시뮬레이션을 활용한 비 마커기반 증강현실 시스템 구현)

  • Sung, Nak-Jun;Choi, Yoo-Joo;Hong, Min
    • Journal of Internet Computing and Services
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    • v.17 no.4
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    • pp.35-42
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    • 2016
  • Recently many researches have been focused on the use of the markerless augmented reality system using face, foot, and hand of user's body to alleviate many disadvantages of the marker based augmented reality system. In addition, most existing augmented reality systems have been utilized rigid objects since they just desire to insert and to basic interaction with virtual object in the augmented reality system. In this paper, unlike restricted marker based augmented reality system with rigid objects that is based in display, we designed and implemented the markerless augmented reality system using deformable objects to apply various fields for interactive situations with a user. Generally, deformable objects can be implemented with mass-spring modeling and the finite element modeling. Mass-spring model can provide a real time simulation and finite element model can achieve more accurate simulation result in physical and mathematical view. In this paper, the proposed markerless augmented reality system utilize the mass-spring model using tetraheadron structure to provide real-time simulation result. To provide plausible simulated interaction result with deformable objects, the proposed method detects and tracks users hand with Kinect SDK and calculates the external force which is applied to the object on hand based on the position change of hand. Based on these force, 4th order Runge-Kutta Integration is applied to compute the next position of the deformable object. In addition, to prevent the generation of excessive external force by hand movement that can provide the natural behavior of deformable object, we set up the threshold value and applied this value when the hand movement is over this threshold. Each experimental test has been repeated 5 times and we analyzed the experimental result based on the computational cost of simulation. We believe that the proposed markerless augmented reality system with deformable objects can overcome the weakness of traditional marker based augmented reality system with rigid object that are not suitable to apply to other various fields including healthcare and education area.

Human Lung Cancer Cell Xenografts Implanted under the Capsule of Kidney, Spleen and Liver (폐암 세포주를 사용한 신, 비장 및 간 피막하 분식법의 비교)

  • 김수현;김종인;이해영;조봉균;박성달;김송명
    • Journal of Chest Surgery
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    • v.36 no.10
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    • pp.711-720
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    • 2003
  • Bakground : Complete resection by the surgery has been selected as the treatment of choice in lung cancer patients, but in cases of recurrence after excision or inoperable cases, the importance of anticancer chemotherapy has been emphasized. If one can select a set of the sensitive chemotherapeutic agents before anticancer chemotherapy, it will give more favourable results. Subrenal capsular assay has been recognized as a useful in-vivo chemosensitivity test of thoracic and abdominal tumors and it can be done in a short time for a rapid interpretation of tumor responsiveness to anticancer chemotherapeutic drugs. It has been reported that various kinds of cancer cells can be implantable to the kidney, but so far there is no comparative study of xenogeneic cell implantation on liver, spleen and kidney. The author implanted the human lung cancer cells under the capsule of S.D rat's liver, spleen and kidney respectively and compared the pattern of growth and histology. Material and Method: After incubation of human lung cancer cell line (SW-900 G IV) in RPMI 1640 (Leibovitz L-15 medium) culture media, 3${\times}$3${\times}$3 mm size fibrin clots which contain 108 cancer cells were made. Thereafter the fibrin clots were implanted at subcapsule area of liver, spleen and kidney of S.D. female rat. For immune suppression, cyclosporin-A (80 mg/Kg) was injected subcutaneously daily from post-implantation first day to sixth day. The body weight was measured at pre and post implantation periods. The growth pattern and the size of tumor mass were observed and the pathologic examination and serum tumor marker tests were performed. Result: Body weight increased in both of control and experimental groups. Serum Cyfra 21-1 was not detected. Serum levels of CEA and NSE revealed no significant change. The SCC-Ag increased significantly in implanted group. The growth rate of human lung cancer cells which was implanted on spleen was higher than on liver or kidney. The surface area, thickness, and volume of tumor mass were predominant at spleen. The success rates of implantation were 80% on kidney, 76.7% on spleen and 43.3% on liver. Pathologic examination of implanted tumors showed characteristic findings according to different organs. Tumors that were implanted on kidney grew in a round shape, small and regular pattern. In the spleen, tumors grew well and microscopic neovascularization and tumor thrombi were also found, but the growth pattern was irregular representing frequent daughter mass. Human lung cancer cells that were implanted in the liver, invaded to the liver parenchyme, and had low success rate of implantation. Microscopically, coagulation necrosis and myxoid fibrous lesion were observed. Conclusion: The success rate of implantation was highest in the kidney. And the mass revealed regular growth that could be measured easily. The SCC-Ag was presented earlier than CEA or Cyfra21-1. The Cyfra21-1 was not detected at early time after implantation. The best model for tumor implantation experiment for chemosensitivity test was subrenal capsular analysis than liver and spleen and the useful serum tumor marker in early period of implantation was the SCC-Ag.

Establishment of the High-Throughput Hair Roots' DNA Isolation System and Verification of Its Appicability for Hanwoo Traceability Using the 11 Microsatellite Makes (대량 모근 시료 DNA 분리 체계 확립과 11 microsatellite maker를 사용하는 한우 생산이력제로의 적용가능성 검증)

  • Lim, Hyun-Tae;Lee, Sang-Ho;Yoo, Chae-Kyoung;Sun, Du-Won;Cho, In-Cheol;Yoon, Du-Hak;Yang, Dae-Young;Cheong, Il-Cheong;Lee, Jung-Gyu;Jeon, Jin-Tae
    • Journal of agriculture & life science
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    • v.44 no.6
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    • pp.91-99
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    • 2010
  • We used a multiplex PCR primer set composed of 11 microsatellite (MS) markers and two sexing markers for gender detection. Genomic DNA extracted from hair roots of 3,510 Hanwoo were genotyped. Based on the 11MS markers, no animals had identical genotypes(TGLA227, BM2113, TGLA53, ETF10, SPS115, TGLA122, ETH3, ETH225, BM1824 and INRA23). The expected probability of identity among genotypes of random individuals (PI), the probability of identity among genotypes from random half-sibs ($PI_{half-sibs}$) and among genotypes of random individuals, and the probability of identity among genotypes from random sibs ($PI_{sibs}$) were estimated as $1.31{\times}10^{-23}$, $2.52{\times}10^{-16}$and $1.09{\times}10^{-6}$, respectively using the API-CALC program, version 1.0. We successfully completed the genotype analysis of 3,510 Hanwoo with a 3.93% genotyping failure rate. It was revealed that extracting DNA from the hair root was a time-efficient and cost-effective method to collect specimens for DNA isolation from live animals. This method also minimized stress for the animals during specimen collection. Among the hair roots from the back, belly, upper tail and lower tail, 5~13 hair roots of the lower tail led to the best genotype analysis results. Finally, we established a 96-well-format method of DNA preparation applicable for high- throughput genotype analysis.

Genome-wide Association Study to Identify Quantitative Trait Loci for Meat and Carcass Quality Traits in Berkshire

  • Iqbal, Asif;Kim, You-Sam;Kang, Jun-Mo;Lee, Yun-Mi;Rai, Rajani;Jung, Jong-Hyun;Oh, Dong-Yup;Nam, Ki-Chang;Lee, Hak-Kyo;Kim, Jong-Joo
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1537-1544
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    • 2015
  • Meat and carcass quality attributes are of crucial importance influencing consumer preference and profitability in the pork industry. A set of 400 Berkshire pigs were collected from Dasan breeding farm, Namwon, Chonbuk province, Korea that were born between 2012 and 2013. To perform genome wide association studies (GWAS), eleven meat and carcass quality traits were considered, including carcass weight, backfat thickness, pH value after 24 hours (pH24), Commission Internationale de l'Eclairage lightness in meat color (CIE L), redness in meat color (CIE a), yellowness in meat color (CIE b), filtering, drip loss, heat loss, shear force and marbling score. All of the 400 animals were genotyped with the Porcine 62K SNP BeadChips (Illumina Inc., USA). A SAS general linear model procedure (SAS version 9.2) was used to pre-adjust the animal phenotypes before GWAS with sire and sex effects as fixed effects and slaughter age as a covariate. After fitting the fixed and covariate factors in the model, the residuals of the phenotype regressed on additive effects of each single nucleotide polymorphism (SNP) under a linear regression model (PLINK version 1.07). The significant SNPs after permutation testing at a chromosome-wise level were subjected to stepwise regression analysis to determine the best set of SNP markers. A total of 55 significant (p<0.05) SNPs or quantitative trait loci (QTL) were detected on various chromosomes. The QTLs explained from 5.06% to 8.28% of the total phenotypic variation of the traits. Some QTLs with pleiotropic effect were also identified. A pair of significant QTL for pH24 was also found to affect both CIE L and drip loss percentage. The significant QTL after characterization of the functional candidate genes on the QTL or around the QTL region may be effectively and efficiently used in marker assisted selection to achieve enhanced genetic improvement of the trait considered.

Development of glufosinate-tolerant GMO detection markers for food safety management (식품안전관리를 위한 제초제 glufosinate 특이적 GM 작물 검출마커 개발)

  • Song, Minji;Qin, Yang;Cho, Younsung;Park, TaeSung;Lim, Myung-Ho
    • Korean Journal of Food Science and Technology
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    • v.52 no.1
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    • pp.40-45
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    • 2020
  • Over 500 genetically modified organisms (GMOs) have been developed since 1996, of which nearly 44% have glufosinate herbicide-tolerant traits. Identification of specific markers that can be used to identify herbicide-tolerant traits is challenging as the DNA sequences of the gene(s) of a trait are highly variable depending on the origin of the gene(s), plant species, and developers. To develop specific PCR marker(s) for the detection of the glufosinate-tolerance trait, DNA sequences of several pat or bar genes were compared and a diverse combination of PCR primer sets were examined using certified reference materials or transgenic plants. Based on both the qualitative and quantitative PCR tests, a primer set specific for pat and non-specific for bar was developed. Additionally, a set of markers that can detect both pat and bar was developed, and the quantitative PCR data indicated that the primer pairs were sensitive enough to detect 0.1% of the mixed seed content rate.