• Title/Summary/Keyword: Specific markers

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Use of RT-PCR for CK20 and CEA mRNA to Detect of Micrometastasis in the Draining Venous Blood and the Peritoneum in Gastric Cancer Patients (위암에서 CK20과 CEA의 RT-PCR을 이용한 수술조작 전후 유출 정맥과 복막에서의 미세전이의 검출)

  • Joo Jai Kyun;Lee Ji Hee;Koh Yang Seok;Kim Jung Chul;Ryu Seong Yeob;Kim Dong Yi;Kim Young Jin
    • Journal of Gastric Cancer
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    • v.3 no.3
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    • pp.128-133
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    • 2003
  • Purpose: The benefits of the 'no-touch' isolation techniquethat is usually performed to prevent the circulation of tumor cells are not evident. The aim of this study was to determine whether the no-touch isolation technique for treating gastrointestinal cancers could prevent the circulation of tumor cells detected by reverse transcriptase polymerase chain reaction (RT-PCR). Matrials and Methods: By using RT-PCR to amplify mRNAs for two specific epithelial markers, carcinoembryonic antigen (CEA) and cytokeratin 20 (CK-20), we examined 34 gastric cancer patients who had been histologically diagnosed and 22 patients had undergone serosal and peritoneal brushing. Results: In 10 ($29.4\%$) of the 34 gastric cancer patients, we detected CK20 mRNA before manipulation, and in 17 ($51.5\%$) of those patients, after we detected it. The density of the CK20 mRNA band was increased in 11 cases ($33.3\%$) and the density was decreased in 2 cases ($6.1\%$). In 16 ($48.5\%$) of the 34 gastric cancer patients, we detected CEA mRNA before manipulation, and in 17 ($51.5\%$) patients after we detected it. The density of the CEA mRNA band was increased in 8 cases ($24.2\%$) and decreased in 3 cases ($9.1\%$). Conclusion: These result suggest that the ' no-touch isolation technique ' might be useful when operating on advanced gastric cancer patients and that serosal or Douglas pouch brushing can be used to determine the status of micrometastasis.

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QTL Analysis of Rice Heading-related Genes Using Cheongcheong/Nagdong Doubled Haploid Genetic Map (청청/낙동 배가반수체 유전자 지도를 이용한 쌀의 출수기 관련 양적형질유전자좌(QTL) 분석)

  • Jang, Yoon-Hee;Park, Jae-Ryoung;Kim, Kyung-Min
    • Journal of Life Science
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    • v.30 no.10
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    • pp.844-850
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    • 2020
  • Disaster-related extreme weather is rapidly increasing due to climate change. In Korea, typhoons accompanied by rainfall usually approach in August and September, causing great damage. The purpose of this study is to find a gene that regulates the heading date of rice in order to avoid loss of harvest from climate change and typhoons. Cheongcheong/Nagdong doubled haploid (CNDH) was used as the plant material to investigate the location of heading-related genes using QTL and sequence analysis by cloning the gene. In the distribution chart, the heading dates, culm lengths, panicle lengths, numbers of panicles, and 1,000-grain weights all have normal distributions. QTL analysis found 13 contigs on chromosome 8. One QTL, named qHd8, was detected on chromosome 8. The range at qHd8 was approximately 7.7 cM, with RM72 and RM404 markers near the peak. There were 13 contigs and 1 ORF. Protein sequence analysis showed that rice was similar to Os08g0341700, AtSFH13, and AtSFH7 proteins. Os08g0341700, which is involved in signal transduction, is similar to phosphatidylinositol transfer-like protein II, and complete information is not available, but it is believed to play a role in the phosphatidylinositol-specific signaling pathway related to Sec14P.

Detection of Clavibacter michiganensis subsp. michiganensis Assisted by Micro-Raman Spectroscopy under Laboratory Conditions

  • Perez, Moises Roberto Vallejo;Contreras, Hugo Ricardo Navarro;Herrera, Jesus A. Sosa;Avila, Jose Pablo Lara;Tobias, Hugo Magdaleno Ramirez;Martinez, Fernando Diaz-Barriga;Ramirez, Rogelio Flores;Vazquez, Angel Gabriel Rodriguez
    • The Plant Pathology Journal
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    • v.34 no.5
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    • pp.381-392
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    • 2018
  • Clavibacter michiganensis subsp. michiganesis (Cmm) is a quarantine-worthy pest in $M{\acute{e}}xico$. The implementation and validation of new technologies is necessary to reduce the time for bacterial detection in laboratory conditions and Raman spectroscopy is an ambitious technology that has all of the features needed to characterize and identify bacteria. Under controlled conditions a contagion process was induced with Cmm, the disease epidemiology was monitored. Micro-Raman spectroscopy ($532nm\;{\lambda}$ laser) technique was evaluated its performance at assisting on Cmm detection through its characteristic Raman spectrum fingerprint. Our experiment was conducted with tomato plants in a completely randomized block experimental design (13 plants ${\times}$ 4 rows). The Cmm infection was confirmed by 16S rDNA and plants showed symptoms from 48 to 72 h after inoculation, the evolution of the incidence and severity on plant population varied over time and it kept an aggregated spatial pattern. The contagion process reached 79% just 24 days after the epidemic was induced. Micro-Raman spectroscopy proved its speed, efficiency and usefulness as a non-destructive method for the preliminary detection of Cmm. Carotenoid specific bands with wavelengths at 1146 and $1510cm^{-1}$ were the distinguishable markers. Chemometric analyses showed the best performance by the implementation of PCA-LDA supervised classification algorithms applied over Raman spectrum data with 100% of performance in metrics of classifiers (sensitivity, specificity, accuracy, negative and positive predictive value) that allowed us to differentiate Cmm from other endophytic bacteria (Bacillus and Pantoea). The unsupervised KMeans algorithm showed good performance (100, 96, 98, 91 y 100%, respectively).

Circulating miR-195 as a Therapeutic Biomarker in Turkish Breast Cancer Patients

  • Cecener, Gulsah;Ak, Secil;Eskiler, Gamze Guney;Demirdogen, Elif;Erturk, Elif;Gokgoz, Sehsuvar;Polatkan, Volkan;Egeli, Unal;Tunca, Berrin;Tezcan, Gulcin;Topal, Ugur;Tolunay, Sahsine;Tasdelen, Ismet
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.9
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    • pp.4241-4246
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    • 2016
  • Background: Dysregulation of miRNA expression may be used as a biomarker for specific tumours because it may contribute to development of cancer. Circulating miRNA profiles have been highlighted for their potential as predictive markers in heterogeneous diseases such as breast cancer. In the literature, there is evidence that miR-195 levels are differentially expressed pre- and post-operative periods in breast cancer patients. At the same time, miRNA expression levels may vary because of ethnic origins. This study aimed to determine expression levels and potential roles of miR-195 in Turkish breast cancer patients. Materials and Methods: The expression patterns of miR-195 were initially examined in breast cancer tissues (luminal A and B type) (n=96). Subsequently, blood samples were prospectively collected from preoperative and postoperative Turkish breast cancer patients and disease free controls. Total RNA was isolated, and the expression level of miR-195 was quantified by real-time PCR. Results: We found that miR-195 level was altered in Turkish breast cancer patients, with down-regulation evident in breast cancer tissues compared to normal adjacent specimens. Furthermore, circulating levels of miR-195 was significantly decreased in post-operative blood samples compared with pre-operative levels (p=0.01 and <0.05). However, miR-195 was significantly increased in pre-operative blood samples of the luminal B type (p=0.04 and <0.05). Conclusions: This study represents the first report of a miR-195 expression profile in Turkish breast cancer patients. Our data suggests that miR-195 levels might be a clinically useful biomarker in the earliest stage of Turkish breast cancer patients.

The Study on Vitrification and Ultrarapid Thawing of Human Embryonic Stem Cells (인간 배아 줄기세포의 초자화 동결 및 초급속 융해에 관한 연구)

  • Moon, Shin-Yong;Park, Yong-Bin;Kim, Hee-Sun;Sung, Ki-Chung;Oh, Sun-Kyung;Chun, Dae-Woo;Suh, Chang-Suk;Choi, Young-Min;Kim, Jung-Gu;Lee, Jin-Yong;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.1
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    • pp.13-20
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    • 2002
  • Objective: This study was carried out to establish the effectiveness of the vitrification method and the optimal cryoprotectants in the cryopreservation of human embryonic stem cells (ESC). Materials and Methods: Human ESC clumps established at Seoul National University Hospital (SNUhES 1) were cryopreserved with the vitrification method using the EM grid. EDS and EFS40 were used as vitrification solutions. Results: Between the EDS and EFS40 groups, there was no significant difference in the recovery rate after cryopreservation of human ESC. The formation rates of ESC colonies in the vitrified groups were significantly lower than those in the control ESC group (p<0.05, p<0.05). In addition, the formation rate of ESC colonies in the EDS group was significantly higher than that in the EFS40 group (p<0.05). The ESC colonies in the vitrified groups were significantly smaller after culture duration of 2 and 4 days, respectively, compared with the control ESC group (p<0.1, p<0.05). However, these effects could be reduced to nonsignificant level by the additional culture of ESC colonies. The vitrified human ESC retained the properties of pluripotent cells, including the expression of cell surface. markers for the undifferentiated cells such as alkaline phosphatase and SSEA-4 (stage-specific embryonic antigen-4), and the expression of transcription factor Oct-4 (octamer-binding transcription factor-4), and the normal karyotype. Conclusion: The vitrification method using the EM grid and EDS solution was confirmed to be very effective for the cryopreservation of human ESC.

Phosphatidylserine Enhances Skin Barrier Function Through Keratinocyte Differentiation (포스파티딜세린의 각질세포 분화 유도를 통한 피부장벽 기능 강화)

  • Chung, So-Young;Nam, Sang-June;Choi, Wang-Keun;Seo, Mi-Young;Kim, Jin-Wook;Lee, Seung-Hun;Park, Chang-Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.1 s.55
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    • pp.17-22
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    • 2006
  • Phosphatidyiserine (PS) is a phospholipid which plays the structural role in membranes and serves as a cofactor of signaling enzymes for diverse cellular functions. In this study, we observed that topical treatment with PS significantly decreased trans-epidermal water loss (TEWL) induced by tape-stripping in hairless mice. Also, ceramides in epidermis were increased in PS-treated group compared to vehicle-treated one in vivo. the amounts of non-hydroxyl ceramide (NHCER) (1.4 fold) and glucosylceramide (glucosylCER) (1.6 fold), in the skin of hairless mice, were increased by topical treament with PS. Also, we demonstrated that PS stimulated keratinocyte differentiation. We observed that PS treatment morphologically altered normal human keratinocyte (NHK) from the proliferating phase to the differentiating one, suggesting that PS stimulated epidermal differentiation in NHK. We also showed that the expressions of the specific markers for epidermal differentiation, involucrin (INV) (3.5 fold up) and transglutaminase 1 (TG'ase 1) (3 fold up), were significantly increased by PS treatment, compared to untreated control in vitro. In addition, topical treatment with PS resulted in a progressive increase in INV and loricrin protein levels in vivo. In conclusion, we provide the first evidence for the physiological activities of PS in skin, and we suggest that PS strengthen the epidermal permeability harrier by stimulation of keratinocyte differentiation.

A Diagnostic Algorithm after Newborn Screening for Hypermethioninemia (고메티오닌혈증의 신생아 선별 검사 후 진단 알고리즘)

  • Kim, Yoo-Mi
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.16 no.1
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    • pp.1-9
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    • 2016
  • Newborn screening (NBS) is important if early intervention is effective in a disorder and if there are sensitive and specific biochemical markers to detect disorder. Methionine is a useful marker to detect abnormal methionine-homocysteine metabolism, especially homocystinuria which needs urgent medical intervention. However, hypermethioninemia could occur in other metabolic disorder including liver disease, tyrosinemia type I, methionine adenosyltransferase (MAT) I/III deficiency, glycine N-methyltransferase (GNMT) deficiency, or adenosylhomocysteine hydrolase deficiency. However, experience with NBS for homocystinurias and methylation disorders is limited. Especially, MAT I/III deficiency which is the most common cause of persistent hypermethioninemia have two inheritance, autosomal recessive (AR) and autosomal dominant (AD), and their clinical manifestation is different between AR and AD. Here, author reviewed recent articles of guideline and proposed guideline for homocystinuria and methylation disorder.

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Selective and Random Patterning of Programmed Cell Death in Zebrafish Embryonic Development

  • Hwang, Chang-Nam;Kim, Joon;Lee, Sang-Ho
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.118-118
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    • 2002
  • Programmed cell death (PCD) is thought as a well-controlled process by which unwanted cells are selectively eliminated. During the last decade many researches have elucidated molecules and their interactions involved in cell death by using largely in vitro induction of cell death or survival signals in a more defined manner, While these critical information and novel findings provide us with clearer understanding of mechanisms underlying cell death, it does by no means explain how PCD occurs and which cells or tissues are affected during normal embryonic development in vivo. In this study, we used zebrafish to examine whether the PCD is occurring selectively or randomly in developing embryos by whole mount in situ TUNEL analysis with specific markers for neural cells. The result revealed that the degree and distribution of TUNEL staining varied considerably throughout gastrulation stage, and there was also a number of TUNEL-negative embryos. Most of TUNEL-positive cells were scattered randomly throughout the blastoderm. During the gastrulation stage about 75 % of the embryos analyzed exhibited more than 5 TUNEL-positive cells. As the dorsal epiblast begins to thicken rather abruptly near the end of gastrulation, TUNEL-positive cells were mainly located along the dorsal side. Although there were some variations in TUNEL staining during segmentation and pharyngeal stages, TUNEL staining continued to be localized to the central nervous system, and was also detected in the sensory organs, trigeminal ganglions, and the primary sensory neurons. High levels of the cell death in developing brain between 20-somite and prim-6 stages are thought to play a role in the morphogenesis and organization of the brain. At prim-16 stage, cell death is considerably reduced in the brain region. Dying cells are mainly localized to the prospective brain region where ectodermal cells are about to initiate neurogenesis. As development progressed, high levels and more reproducible patterns of cell death were observed in the developing nervous system. Intensive TUNEL staining was restricted to the trigeminal ganglions, the primary sensory neurons, and sensory organs, such as olfactory pits and otic vesicles. Thus, PCD patterning in zebrafish embryos occurs randomly at early stages and becomes restricted to certain region of the embryos. The spatio-temporal pattern of PCD during the early embryonic development in zebrafish will provide basic information for further studies to elucidate genes involved in. regulation of PCD largely unknown in vivo during vertebrate embryogenesis.

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Evaluation of Digital PCR as a Technique for Monitoring Acute Rejection in Kidney Transplantation

  • Lee, Hyeseon;Park, Young-Mi;We, Yu-Mee;Han, Duck Jong;Seo, Jung-Woo;Moon, Haena;Lee, Yu-Ho;Kim, Yang-Gyun;Moon, Ju-Young;Lee, Sang-Ho;Lee, Jong-Keuk
    • Genomics & Informatics
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    • v.15 no.1
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    • pp.2-10
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    • 2017
  • Early detection and proper management of kidney rejection are crucial for the long-term health of a transplant recipient. Recipients are normally monitored by serum creatinine measurement and sometimes with graft biopsies. Donor-derived cell-free deoxyribonucleic acid (cfDNA) in the recipient's plasma and/or urine may be a better indicator of acute rejection. We evaluated digital PCR (dPCR) as a system for monitoring graft status using single nucleotide polymorphism (SNP)-based detection of donor DNA in plasma or urine. We compared the detection abilities of the QX200, RainDrop, and QuantStudio 3D dPCR systems. The QX200 was the most accurate and sensitive. Plasma and/or urine samples were isolated from 34 kidney recipients at multiple time points after transplantation, and analyzed by dPCR using the QX200. We found that donor DNA was almost undetectable in plasma DNA samples, whereas a high percentage of donor DNA was measured in urine DNA samples, indicating that urine is a good source of cfDNA for patient monitoring. We found that at least 24% of the highly polymorphic SNPs used to identify individuals could also identify donor cfDNA in transplant patient samples. Our results further showed that autosomal, sex-specific, and mitochondrial SNPs were suitable markers for identifying donor cfDNA. Finally, we found that donor-derived cfDNA measurement by dPCR was not sufficient to predict a patient's clinical condition. Our results indicate that donor-derived cfDNA is not an accurate predictor of kidney status in kidney transplant patients.

The Prediction of the Expected Current Selection Coefficient of Single Nucleotide Polymorphism Associated with Holstein Milk Yield, Fat and Protein Contents

  • Lee, Young-Sup;Shin, Donghyun;Lee, Wonseok;Taye, Mengistie;Cho, Kwanghyun;Park, Kyoung-Do;Kim, Heebal
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.1
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    • pp.36-42
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    • 2016
  • Milk-related traits (milk yield, fat and protein) have been crucial to selection of Holstein. It is essential to find the current selection trends of Holstein. Despite this, uncovering the current trends of selection have been ignored in previous studies. We suggest a new formula to detect the current selection trends based on single nucleotide polymorphisms (SNP). This suggestion is based on the best linear unbiased prediction (BLUP) and the Fisher's fundamental theorem of natural selection both of which are trait-dependent. Fisher's theorem links the additive genetic variance to the selection coefficient. For Holstein milk production traits, we estimated the additive genetic variance using SNP effect from BLUP and selection coefficients based on genetic variance to search highly selective SNPs. Through these processes, we identified significantly selective SNPs. The number of genes containing highly selective SNPs with p-value <0.01 (nearly top 1% SNPs) in all traits and p-value <0.001 (nearly top 0.1%) in any traits was 14. They are phosphodiesterase 4B (PDE4B), serine/threonine kinase 40 (STK40), collagen, type XI, alpha 1 (COL11A1), ephrin-A1 (EFNA1), netrin 4 (NTN4), neuron specific gene family member 1 (NSG1), estrogen receptor 1 (ESR1), neurexin 3 (NRXN3), spectrin, beta, non-erythrocytic 1 (SPTBN1), ADP-ribosylation factor interacting protein 1 (ARFIP1), mutL homolog 1 (MLH1), transmembrane channel-like 7 (TMC7), carboxypeptidase X, member 2 (CPXM2) and ADAM metallopeptidase domain 12 (ADAM12). These genes may be important for future artificial selection trends. Also, we found that the SNP effect predicted from BLUP was the key factor to determine the expected current selection coefficient of SNP. Under Hardy-Weinberg equilibrium of SNP markers in current generation, the selection coefficient is equivalent to $2^*SNP$ effect.