• Title/Summary/Keyword: Differential expression analysis

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Gene Expression Profiling by RNA Sequencing in Mature/Immature Oocytes of Chicken (닭의 성숙/미성숙란에서 RNA Sequencing을 이용한 유전자 발현 양상 고찰)

  • Kang, Kyung-Soo;Jang, Hyun-Jun;Park, Mi Na;Choi, Jung-Woo;Chung, Won-Hyong;Heo, Kang-Nyeong;Choe, Chang-Yong;Kim, Young-Joo;Lee, Si-Woo;Cho, Eun-Seok;Kim, Namshin;Kim, Tae-Hun;Han, Jae-Yong;Lee, Kyung-Tai
    • Korean Journal of Poultry Science
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    • v.41 no.4
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    • pp.287-296
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    • 2014
  • Chicken eggs undergo various physiological changes during egg maturation. To study genes associated with the egg maturation in pre-ovulation (immature) and post-ovulation (mature), we compared gene expression patterns between in the immature egg and mature egg using RNA sequencing data. Mature and immature eggs were obtained from a Heuksaek Jaerae-jong of Korean native chicken. Total RNAs obtained from the eggs were sequenced by Illumina HiSeq 2000 platform, and the generated sequence reads were mapped to Galgal4 reference sequence assembly using Tuxedo Protocol. From the comparison of the RNA sequencing data, 315 genes were differentially expressed between mature and immature eggs, and 46 genes were only detected in immature egg. Further gene ontology (GO) analysis was performed for the differentially expressed genes using DAVID, showing that 29 and 28 GO terms were independently clustered from mature and immature, respectively. From those clustered GO terms, genes related to germ cell development, sex differentiation and defense response to bacterium were mainly expressed in the immature egg, while genes related to regulation of apoptosis, steroid metabolic process and lipid homeostasis were mainly detected in the mature egg. Our results could contribute to understand egg maturation before and after ovulation, and develop genetic markers for improving egg quality and productivity.

Identification of the Plant Part of Gleditsia sinensis that Activates Nrf2, an Anti-oxidative Transcription Factor (조협의 부위에 따른 항산화 전사인자 Nrf2 활성 효과)

  • Choi, Jiyeon;Kim, Kyun Ha;Choi, Jun Yong;Han, Chang Woo;Ha, Ki Tae;Jeong, Han-Sol;Joo, Myungsoo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.3
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    • pp.303-309
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    • 2014
  • The fruit of Gleditsia sinensis has been extensively used as a key ingredient of an herbal remedy for the treatment of various inflammatory diseases in traditional Korean Medicine. However, the reason of using the fruit of G. sinensis for the remedy is unclear. Since Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is a key anti-inflammatory transcription factor, which is activated by the fruit of G. sinesis, we examined whether other plant parts of G. sinensis are also capable of suppressing inflammatory responses by activating Nrf2. Water extracts of various parts of G. sinensis were prepared and tested for Nrf2 activation by reporter assay and western blot analysis. Our results show that the hull of G. sinensis is the most potent in activating Nrf2. Sequential organic solvent extraction of the hull show that all the fractions had a higher potency in activating Nrf2 than the water extract, albeit differential degrees. The hull originated from Korea in general activated Nrf2 strongly compared to that of China. Chloroform fraction of the hull was further examined, showing that the fraction induced nuclear localization of Nrf2, indicative of activated Nrf2, and Nrf2-dependent gene expression including NAD(P)H dehydrogenase quinone 1 (NQO-1), glutamate-cysteine ligase catalytic subunit (GCLC), and heme oxygenase - 1 (HO-1). Therefore, our results show that, among other plant parts examined in this study, the hull of G. sinensis is the most potent, providing the experimental basis for the use of the hull of G. sinensis as an active ingredient for an anti-inflammatory remedy.

Cloning of the posterior silk glands specific-expressed gene of silkworm (누에 후부실샘 특이 발현 유전자 클로닝)

  • Piao, Yulan;Kim, Seong-Ryul;Kim, Sung-Wan;Kang, Seok-Woo;Goo, Tae-Won;Choi, Kwang-Ho
    • Journal of Sericultural and Entomological Science
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    • v.53 no.1
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    • pp.44-49
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    • 2015
  • We characterized tissue specific-expressed genes in the posterior silk gland of Bombyx mori using by the Annealing Control Primer based differential display-PCR manner. In this study, we isolated 34 differentially expressed PCR amplicons, which one of these was identified as a novel transcript named as ACP-16 (366 bp), its expression was observed only in the posterior silk glands by Northern blot analysis. To determine promoter region of the ACP-16, we isolated and analyzed a phage DNA having 1.7 kb-long genome DNA including the open reading flame and 5'- upstream untranslated region of the ACP-16 gene from a genomic DNA library. We have estimated a promoter region of the ACP-16 gene by a web promoter prediction engine, which locates -750 ~ -165 from translation initiation site (ATG, +1). ACP-16 gene is necessary to more studies about critical biological role in order to apply the silkworm's transgenic system.

Effects of Differential Distribution of Microvessel Density, Possibly Regulated by miR-374a, on Breast Cancer Prognosis

  • Li, Jian-Yi;Zhang, Yang;Zhang, Wen-Hai;Jia, Shi;Kang, Ye;Tian, Rui
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.1715-1720
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    • 2013
  • Background: The discovery that microRNAs (miRNAs) regulate proliferation, invasion and metastasis provides a principal molecular basis of tumor heterogeneity. Microvessel distribution is an important characteristic of solid tumors, with significant hypoxia occurring in the center of tumors with low blood flow. The distribution of miR-374a in breast tumors was examined as a factor likely to be important in breast cancer progression. Methods: Breast tissue samples from 40 patients with breast cancer were classified into two groups: a highly invasive and metastatic group (HIMG) and a low-invasive and metastatic Group (LIMG). Samples were collected from the center and edge of each tumor. In each group, six specimens were examined by microRNA array, and the remaining 14 specimens were used for real-time RT-qPCR, Western blot and immunohistochemical analyses. Correlation analysis was performed for the miRNAs and target proteins. Follow-up was carried out during 28 months to 68 months after surgery, and survival data were analyzed. Results: In the LIMG, the relative content of miR-374a was lower in the center of the tumor than at its edge; in the HIMG, it was lower at the edge of the tumor, and miR-374a levels were lower in breast cancer tissues than in normal tissues. There was no difference between VEGF-A and VCAM-1 mRNA levels at the edge and center of the tumor; however, we observed a significant difference between VEGF-A and VCAM-1 protein expression levels in these two regions. There was a negative correlation between miR-374a and target protein levels. The microvessel density (MVD) was lower in the center of the tumor than at its edge in HIMG, but the LIMG vessels were uniformly distributed. There was a significant positive correlation between MVD and the number of lymph node metastases (Pearson correlation, r=0.912, P<0.01). The median follow-up time was 48.5 months. LIMG had higher rate of disease-free survival (100%, P=0.013) and longer median survival time (66 months) than HIMG, which had a lower rate of 75% and shorter median survival time (54 months). Conclusions: Our data demonstrated miR-374a to be differentially distributed in breast cancer; VEGF-A and VCAM-1 mRNA had coincident distribution, and the distribution of teh respective proteins was uneven and opposite to that for the miR-374a. These data might explain the differences in the distribution of MVD in breast cancer and variation in breast cancer prognosis.

Identification of Functional and In silico Positional Differentially Expressed Genes in the Livers of High- and Low-marbled Hanwoo Steers

  • Lee, Seung-Hwan;Park, Eung-Woo;Cho, Yong-Min;Yoon, Duhak;Park, Jun-Hyung;Hong, Seong-Koo;Im, Seok-Ki;Thompson, J.M.;Oh, Sung-Jong
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.9
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    • pp.1334-1341
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    • 2007
  • This study identified hepatic differentially expressed genes (DEGs) affecting the marbling of muscle. Most dietary nutrients bypass the liver and produce plasma lipoproteins. These plasma lipoproteins transport free fatty acids to the target tissue, adipose tissue and muscle. We examined hepatic genes differentially expressed in a differential-display reverse transcription-polymerase chain reaction (ddRT-PCR) analysis comparing high- and low-marbled Hanwoo steers. Using 60 arbitrary primers, we found 13 candidate genes that were upregulated and five candidate genes that were downregulated in the livers of high-marbled Hanwoo steers compared to low-marbled individuals. A BLAST search for the 18 DEGs revealed that 14 were well characterized, while four were not annotated. We examined four DEGs: ATP synthase F0, complement component CD, insulin-like growth factor binding protein-3 (IGFBP3) and phosphatidylethanolamine binding protein (PEBP). Of these, only two genes (complement component CD and IGFBP3) were differentially expressed at p<0.05 between the livers of high- and low-marbled individuals. The mean mRNA levels of the PEBP and ATP synthase F0 genes did not differ significantly between the livers of high- and low-marbled individuals. Moreover, these DEGs showed very high inter-individual variation in expression. These informative DEGs were assigned to the bovine chromosome in a BLAST search of MS marker subsets and the bovine genome sequence. Genes related to energy metabolism (ATP synthase F0, ketohexokinase, electron-transfer flavoprotein-ubiquinone oxidoreductase and NADH hydrogenase) were assigned to BTA 1, 11, 17, and 22, respectively. Syntaxin, IGFBP3, decorin, the bax inhibitor gene and the PEBP gene were assigned to BTA 3, 4, 5, 5, and 17, respectively. In this study, the in silico physical maps provided information on the specific location of candidate genes associated with economic traits in cattle.

Effect of Long Term Reverse Feeding on the Reproductive and Non-reproductive Tissues in Male Mice

  • Go, Eun Hye;Lee, Sung-Ho
    • Development and Reproduction
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    • v.18 no.3
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    • pp.161-166
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    • 2014
  • Previously, we demonstrated that the shift and/or restriction of feeding time during relatively short-term period (4 weeks) could alter the pituitary gonadotropin expression and the weights of seminal vesicle and prostate in rats. We also found that the reverse feeding (RF) schedule (up to 8 weeks) might induce an adaptable metabolic stress and cause impairment of androgen-dependent reproductive tissues. In the present study, we extended the RF time regimen up to 12 weeks, and measured the reproductive tissue weights. After 4 and 8 weeks of RF, the weights of epididymis were not significantly different. After 12 weeks, however, epididymis weights of RF animals were significantly different (CON 12W : RF 12W = $48.26{\pm}0.62mg$ : $44.05{\pm}1.57mg$, p<0.05). After 4 and 12 weeks of feeding, seminal vesicle weights of RF animals were significantly decreased (CON 4W : RF 4W = $79.36{\pm}8.34mg$ : $46.28{\pm}2.43mg$, p<0.001; CON 12W : RF 12W = $72.04{\pm}3.76mg$ : $46.71{\pm}2.27mg$, p<0.001, respectively). Prostate weights were not changed by RF. Kidney and spleen weights of RF animals were significantly different on weeks 4 and 12 (Kidney, CON 4W : RF 4W = $249.72{\pm}4.20mg$ : $228.41{\pm}3.03mg$, p<0.001; CON 12W : RF 12W = $309.15{\pm}7.49mg$ : $250.72{\pm}6.13mg$, p<0.001, respectively, Spleen, CON 4W : RF 4W = $111.26{\pm}3.76mg$ : $96.88{\pm}4.69mg$, p<0.05; CON 12W : RF 12W = $123.93{\pm}10.72mg$ : $94.68{\pm}5.65mg$, p<0.05, respectively). Histology analysis of seminal vesicle revealed that the thinner epithelial cell layers, reduced complexities of swollen papilla folding in the exocrine glands on weeks 4 and 12 of RF. There was no histological difference between control and RF group on week 8. The present study indicates that up to 12 weeks RF induced differential changes in tissue weights of male mice. In particular, seminal vesicle, kidney and spleen seemed to temporarily adapted to the RF-induced metabolic stress on week 8 of feeding schedule. These results confirmed the our previous study that the RF might induce an adaptable metabolic stress and cause impairment of androgen-dependent reproductive tissues such as epididymis and seminal vesicle as well as non-reproductive tissues such as kidney and spleen. Further studies will be needed to achieve a better understanding of the how does mealtime shift affect the reproductive function and exact nature of adaptation.

Evaluation of Biological Effects of Low Concentrations of Mercury Chloride (II) and Ionizing Radiation in the Prepubertal Male Rats (미성숙 웅성 흰쥐를 이용한 이온화 방사선 조사 및 저농도 염화수은(II)의 음용에 따른 위해성 비교 평가)

  • Kim Ji Hyang;Kim Jin Kyu;Yoon Yong Dal
    • Korean Journal of Environmental Biology
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    • v.22 no.3
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    • pp.411-418
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    • 2004
  • Mercury, one of the most diffused and hazardous organ-specific environmental contaminants, exists in a wide variety of physical and chemical states. The murcury with the nature which evaporates easily can cause an acute or chronic mercury poisoning to workers at mercury-handling workplaces. Although many studies indicate that mercury induces a deleterious damage, little has been reported from the investigations of mercury effects at surrounding levels in living things. The purpose of this study was to evaluate the biological effects of mercury chloride and ionizing radiation. Prepubertal male F344 rats were administered mercury chloride in drinking water throughout the experimental period or were given wholebody irradiation with a dose of 6.5 Gy. The amount changed of body weight during the experimental period showed a 4.9% rise in the mercury-treated group and 14.4% decline in the irradiated group compared with the level of the control group. The results of hematological analysis (red blood cells, white blood cells, hemoglobin, and hematocrit) indicated the differential effects of mercury chloride and ionizing radiation. However the concentration of cortisol as assessed by radioimmunoassay increased in both of the groups. Relative expressions of mRNA related to mitochondrion-mediated apoptosis were investigated using semiquantitative reverse transcription polymerase chain reaction on gonad and urinary organs of the experimental groups. While the expression of Bcl-2 mRNA exhibited different patterns depending on the organs or the experimental groups, both of the experimental groups showed a conspicuous expressions of Bax mRNA. In conclusion, the target organ of mercury chloride seems to be a urinary organ and the pattern of damage induced by mercury chloride differs from that by ionizing radiation.

Preparation and Release Behavior of Ipriflavone-Loaded PLGA Microsphere for Tissue Engineered Bone (이프리플라본을 함유한 생분해성 PLGA 미립구의 제조 및 조직공학적 골재생을 위한 영향평가)

  • So, Jung-Won;Jang, Ji-Wook;Kim, Soon-Hee;Kim, Geun-Ah;Choi, Jin-Hee;Rhee, John-M.;Son, Young-Suk;Min, Byoung-Hyun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.33 no.1
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    • pp.26-32
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    • 2009
  • The aim of this research was to prepare microparticulate systems based on poly (lactide-co-glycolide)(PLGA) for the local release of ipriflavone in order to reduce bone loss. We developed the IP loaded PLGA microspheres using relatively simple oil-in-water(O/W) solvent evaporation method. HPLC was used to perform the in vitro release test of IP and morphology of cell attached on the micro-spheres was investigated using SEM. Cytotoxicity was assayed by cell counting kit-8 (CCK-8) test. Osteogenic differential cells were analyzed by ALP activity. Through RT-PCR analysis, we observed osteocalcin, ALP, and Type I collagen mRNA expression. The release of IP in vitro was more prolonged over 42 days and IP/PLGA microspheres showed the improvement on the cell proliferation, ALP activity and RT-PCR comparing with control (only PLGA). This initial research will be used to direct future work involved in developing this composite injectable bone tissue engineering system.

Identification and Validation of Circulating MicroRNA Signatures for Breast Cancer Early Detection Based on Large Scale Tissue-Derived Data

  • Yu, Xiaokang;Liang, Jinsheng;Xu, Jiarui;Li, Xingsong;Xing, Shan;Li, Huilan;Liu, Wanli;Liu, Dongdong;Xu, Jianhua;Huang, Lizhen;Du, Hongli
    • Journal of Breast Cancer
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    • v.21 no.4
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    • pp.363-370
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    • 2018
  • Purpose: Breast cancer is the most commonly occurring cancer among women worldwide, and therefore, improved approaches for its early detection are urgently needed. As microRNAs (miRNAs) are increasingly recognized as critical regulators in tumorigenesis and possess excellent stability in plasma, this study focused on using miRNAs to develop a method for identifying noninvasive biomarkers. Methods: To discover critical candidates, differential expression analysis was performed on tissue-originated miRNA profiles of 409 early breast cancer patients and 87 healthy controls from The Cancer Genome Atlas database. We selected candidates from the differentially expressed miRNAs and then evaluated every possible molecular signature formed by the candidates. The best signature was validated in independent serum samples from 113 early breast cancer patients and 47 healthy controls using reverse transcription quantitative real-time polymerase chain reaction. Results: The miRNA candidates in our method were revealed to be associated with breast cancer according to previous studies and showed potential as useful biomarkers. When validated in independent serum samples, the area under curve of the final miRNA signature (miR-21-3p, miR-21-5p, and miR-99a-5p) was 0.895. Diagnostic sensitivity and specificity were 97.9% and 73.5%, respectively. Conclusion: The present study established a novel and effective method to identify biomarkers for early breast cancer. And the method, is also suitable for other cancer types. Furthermore, a combination of three miRNAs was identified as a prospective biomarker for breast cancer early detection.

System Development for Measuring Group Engagement in the Art Center (공연장에서 다중 몰입도 측정을 위한 시스템 개발)

  • Ryu, Joon Mo;Choi, Il Young;Choi, Lee Kwon;Kim, Jae Kyeong
    • Journal of Intelligence and Information Systems
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    • v.20 no.3
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    • pp.45-58
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    • 2014
  • The Korean Culture Contents spread out to Worldwide, because the Korean wave is sweeping in the world. The contents stand in the middle of the Korean wave that we are used it. Each country is ongoing to keep their Culture industry improve the national brand and High added value. Performing contents is important factor of arousal in the enterprise industry. To improve high arousal confidence of product and positive attitude by populace is one of important factor by advertiser. Culture contents is the same situation. If culture contents have trusted by everyone, they will give information their around to spread word-of-mouth. So, many researcher study to measure for person's arousal analysis by statistical survey, physiological response, body movement and facial expression. First, Statistical survey has a problem that it is not possible to measure each person's arousal real time and we cannot get good survey result after they watched contents. Second, physiological response should be checked with surround because experimenter sets sensors up their chair or space by each of them. Additionally it is difficult to handle provided amount of information with real time from their sensor. Third, body movement is easy to get their movement from camera but it difficult to set up experimental condition, to measure their body language and to get the meaning. Lastly, many researcher study facial expression. They measures facial expression, eye tracking and face posed. Most of previous studies about arousal and interest are mostly limited to reaction of just one person and they have problems with application multi audiences. They have a particular method, for example they need room light surround, but set limits only one person and special environment condition in the laboratory. Also, we need to measure arousal in the contents, but is difficult to define also it is not easy to collect reaction by audiences immediately. Many audience in the theater watch performance. We suggest the system to measure multi-audience's reaction with real-time during performance. We use difference image analysis method for multi-audience but it weaks a dark field. To overcome dark environment during recoding IR camera can get the photo from dark area. In addition we present Multi-Audience Engagement Index (MAEI) to calculate algorithm which sources from sound, audience' movement and eye tracking value. Algorithm calculates audience arousal from the mobile survey, sound value, audience' reaction and audience eye's tracking. It improves accuracy of Multi-Audience Engagement Index, we compare Multi-Audience Engagement Index with mobile survey. And then it send the result to reporting system and proposal an interested persons. Mobile surveys are easy, fast, and visitors' discomfort can be minimized. Also additional information can be provided mobile advantage. Mobile application to communicate with the database, real-time information on visitors' attitudes focused on the content stored. Database can provide different survey every time based on provided information. The example shown in the survey are as follows: Impressive scene, Satisfied, Touched, Interested, Didn't pay attention and so on. The suggested system is combine as 3 parts. The system consist of three parts, External Device, Server and Internal Device. External Device can record multi-Audience in the dark field with IR camera and sound signal. Also we use survey with mobile application and send the data to ERD Server DB. The Server part's contain contents' data, such as each scene's weights value, group audience weights index, camera control program, algorithm and calculate Multi-Audience Engagement Index. Internal Device presents Multi-Audience Engagement Index with Web UI, print and display field monitor. Our system is test-operated by the Mogencelab in the DMC display exhibition hall which is located in the Sangam Dong, Mapo Gu, Seoul. We have still gotten from visitor daily. If we find this system audience arousal factor with this will be very useful to create contents.