• Title/Summary/Keyword: Differential expression analysis

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Endophytic bacterium Pseudomonas fluorescens strain EP103 was effective against Phytophthora capsici causing blight in chili pepper (식물근권에서 분리한 Pseudomonas fluorescens strain EP103에 의한 고추역병억제)

  • Kim, Tack-Soo;Dutta, Swarnalee;Lee, Se Won;Park, Kyungseok
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.422-428
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    • 2014
  • Endophytic bacterial strains from root tissue of strawberry were screened for their efficacy in growth improvement and control of Phytophthora blight disease of chili pepper plant under greenhouse condition. Plants treated with the strain EP103, identified as Pseudomonas fluorescens, showed growth improvement in terms of fresh weight and root length compared to the untreated control and other endophytic strains. When challenged with Phytophthora capsici, there was significant reduction of disease in EP103 treated plants with an efficacy of 78.7%. There was no direct inhibition of the target pathogen by EP103 when tested under in vitro antibiosis assay. Analysis of differential expression of selected marker genes for induced systemic resistance (ISR) in plants treated with EP103 and challenged with P. capsici showed up-regulation of PR1 and PR10 pathogenesis-related (PR) proteins. PCR analysis showed that EP103 produced secondary metabolites such as pyoluteorin, pyrrolnitrin, hydrogen cyanide and orfamide A. This study indicated the potential of endophytic P. fluorescens strain EP103 as an efficient biocontrol agent against P. capsici in chili pepper plant.

Differential Potential of Stem Cells Following Their Origin - Subacromial Bursa, Bone Marrow, Umbilical Cord Blood - (줄기세포의 분화능의 기원에 따른 비교 - 견봉하 점액낭, 골수, 탯줄 혈액 -)

  • Sim, Sung Woo;Moon, Young Lae;Kang, Jung Hun
    • Clinics in Shoulder and Elbow
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    • v.15 no.2
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    • pp.65-72
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    • 2012
  • Purpose: To evaluate the differentiation potential of stem cells and their immunophenotype from 3 different sources. Methods: Our study involved three stem cell sources-subacromial bursal tissue, bone marrow, and umbilical cord blood. We obtained the subacromial bursal tissue and bone marrow from the patients undergoing shoulder surgery. After collecting the sample, we applied specific induction media for neurogenic, adipogenic and osteogenic differentiation. Also, flow-cytometry analysis was done to reveal the cell surface antigens. Results: We obtained 100% (8 cases) neural and adipogenic differentiation, but 62.5% (5 of 8 cases) osseous differentiation among the subacromial bursal tissue group. Bone marrow derived cells showed 100% neural (6 cases) and adipogenic (5 cases) differentiation, but 80% (4 of 5 cases) osseous differentiation. Umbilical cord blood derived cells revealed 97% (65 of 67 cases) neural, 53.7% (29 of 54 cases) adipogenic and 68.4% (39 of 57 cases) osseous differentiation. Immunophenotype analysis revealed that surface markers of bone marrow, subacromial bursal cell and umbilical cord blood derived mesenchymal stem cells are different from each other. Conclusions: Mesenchymal stem cells are potential agents in regenerative medicine and are characterized by expression of surface markers and by their differentiation potential. Our study with stem cells from subacromial bursal tissue, bone marrow and umbilical cord discovered that each stem cell has unique differentiation potential and function based on its origin. Various stem cells show multi-lineage differentiations in vitro which can be correlated to in vivo conditions.

Real-Time PCR Analysis of Metabolic Pathway of PHB in Acidiphilium cryptum DX1-1

  • Xu, Ai-Ling;Xia, Jin-Lan;Liu, Ke-Ke;Li, Li;Yang, Yu;Nie, Zhen-Yuan;Qiu, Guan-Zhou
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.71-77
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    • 2010
  • The time, yield, and related genes expression of PHB accumulation of Acidiphilium cryptum DX1-1 were investigated under four different initial C/N ratios, 1.2, 2.4, 7.5, and 24. The results of time and yield of poly-$\beta$-hydroxybutyrate (PHB) accumulation show that the initial C/N ratio of 2.4 was optimum for strain DX1-1 to accumulate PHB, but both higher and lower initial C/N ratios did not favor that process. Based on the genome of Acidiphilium cryptum JF-5, 13 PHB accumulation related genes in strain JF-5 were chosen and successfully cloned from strain DX1-1. The differential expressions of the 13 functional genes, in different C/N ratios as cited above, were then studied by real-time PCR. The results show that all the 13 genes were most upregulated when the initial C/N ratio was 2.4, and among which the gene Acry_3030 encoding poly-$\beta$-hydroxybutyrate polymerase and Aery_0626 encoding acetyl-CoA synthetase were much more upregulated than the other genes, which proved that they play the most important role for PHB accumulation, and acetate is the main initial substance for PHB accumulation for strain DX1-1. Potential regulatory motifs analysis showed that the genes related to PHB accumulation are regulated by different promoters and that the motif had weak similarity to the model promoters, suggesting that PHB metabolism in Acidiphilium cryptum may be mediated by a different mechanism.

Identification of Differentially Expressed Radiation-induced Genes in Cervix Carcinoma Cells Using Suppression Subtractive Hybridization (자궁경부암세포에서 방사선조사시 차등 발현되는 유전자 동정)

  • Kim Jun-Sang;Lee Young-Sook;Lee Jeung Hoon;Lee Woong-Hee;Seo Eun Young;Cho Moon-June
    • Radiation Oncology Journal
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    • v.23 no.1
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    • pp.43-50
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    • 2005
  • Purpose : A number of genes and their products are Induced early or late following exposure of cells to ionizing radiation. These radiation-Induced genes have various effects on irradiated cells and tissues. Suppression subtractive hybridization (SSH) based on PCR was used to Identify the differentially expressed genes by radiation in cervix carcinoma cells. Materials and Methods : Total RNA and poly $(A)^+$ mRNA were Isolated from Irradiated and non-irradiated HeLa cells. Forward- and reverse-subtracted cDNA libraries were constructed using SSH. Eighty-eight clones of each were used to randomly select differentially expressed genes using reverse Northern blotting (dot blot analysis). Northern blotting was used to verify the screened genes. Results : Of the 17t clones, 10 genes in the forward-subtracted library and 9 genes In the reverse-subtracted library were identified as differentially expressed radiation-induced genes by PCR-select differential screening. Three clones from the forward-subtracted library were confirmed by Northern blotting, and showed increased expression in a dose-dependent manner, including a telomerase catalytic subunit and sodium channel-like protein gene, and an ESTs (expressed sequence tags) gene. Conclusion : We Identified differentially expressed radiation-induced genes with low-abundance genes with SSH, but further characterization of theses genes are necessary to clarify the biological functions of them.

A systematic exploration of ginsenoside Rg5 reveals anti-inflammatory functions in airway mucosa cells

  • Hyojin Heo;Yumin Kim;Byungsun Cha;Sofia Brito;Haneul Kim;Hyunjin Kim;Bassiratou M. Fatombi;So Young Jung;So Min Lee;Lei Lei;Sang Hun Lee;Geon-woo Park;Byeong-Mun Kwak;Bum-Ho Bin;Ji-Hwan Park;Mi-Gi Lee
    • Journal of Ginseng Research
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    • v.47 no.1
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    • pp.97-105
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    • 2023
  • Background: Hyperactivated airway mucosa cells overproduce mucin and cause severe breathing complications. Here, we aimed to identify the effects of saponins derived from Panax ginseng on inflammation and mucin overproduction. Methods: NCI-H292 cells were pre-incubated with 16 saponins derived from P. ginseng, and mucin overproduction was induced by treatment with phorbol 12-myristate 13-acetate (PMA). Mucin protein MUC5AC was quantified by enzyme-linked immunosorbent assay, and mRNA levels were analyzed using quantitative polymerase chain reaction (qPCR). Moreover, we performed a transcriptome analysis of PMA-treated NCI-H292 cells in the absence or presence of Rg5, and differential gene expression was confirmed using qPCR. Phosphorylation levels of signaling molecules, and the abundance of lipid droplets, were measured by western blotting, flow cytometry, and confocal microscopy. Results: Ginsenoside Rg5 effectively reduced MUC5AC secretion and decreased MUC5AC mRNA levels. A systematic functional network analysis revealed that Rg5 upregulated cholesterol and glycerolipid metabolism, resulting in the production of lipid droplets to clear reactive oxygen species (ROS), and modulated the mitogen-activated protein kinase and nuclear factor (NF)-kB signaling pathways to regulate inflammatory responses. Rg5 induced the accumulation of lipid droplets and decreased cellular ROS levels, and N-acetyl-ⳑ-cysteine, a ROS inhibitor, reduced MUC5AC secretion via Rg5. Furthermore, Rg5 hampered the phosphorylation of extracellular signal-regulated kinase and p38 proteins, affecting the NF-kB signaling pathway and pro-inflammatory responses. Conclusion: Rg5 alleviated inflammatory responses by reducing mucin secretion and promoting lipid droplet-mediated ROS clearance. Therefore, Rg5 may have potential as a therapeutic agent to alleviate respiratory disorders caused by hyperactivation of mucosa cells.

A Study of the Reactive Movement Synchronization for Analysis of Group Flow (그룹 몰입도 판단을 위한 움직임 동기화 연구)

  • Ryu, Joon Mo;Park, Seung-Bo;Kim, Jae Kyeong
    • Journal of Intelligence and Information Systems
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    • v.19 no.1
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    • pp.79-94
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    • 2013
  • Recently, the high value added business is steadily growing in the culture and art area. To generated high value from a performance, the satisfaction of audience is necessary. The flow in a critical factor for satisfaction, and it should be induced from audience and measures. To evaluate interest and emotion of audience on contents, producers or investors need a kind of index for the measurement of the flow. But it is neither easy to define the flow quantitatively, nor to collect audience's reaction immediately. The previous studies of the group flow were evaluated by the sum of the average value of each person's reaction. The flow or "good feeling" from each audience was extracted from his face, especially, the change of his (or her) expression and body movement. But it was not easy to handle the large amount of real-time data from each sensor signals. And also it was difficult to set experimental devices, in terms of economic and environmental problems. Because, all participants should have their own personal sensor to check their physical signal. Also each camera should be located in front of their head to catch their looks. Therefore we need more simple system to analyze group flow. This study provides the method for measurement of audiences flow with group synchronization at same time and place. To measure the synchronization, we made real-time processing system using the Differential Image and Group Emotion Analysis (GEA) system. Differential Image was obtained from camera and by the previous frame was subtracted from present frame. So the movement variation on audience's reaction was obtained. And then we developed a program, GEX(Group Emotion Analysis), for flow judgment model. After the measurement of the audience's reaction, the synchronization is divided as Dynamic State Synchronization and Static State Synchronization. The Dynamic State Synchronization accompanies audience's active reaction, while the Static State Synchronization means to movement of audience. The Dynamic State Synchronization can be caused by the audience's surprise action such as scary, creepy or reversal scene. And the Static State Synchronization was triggered by impressed or sad scene. Therefore we showed them several short movies containing various scenes mentioned previously. And these kind of scenes made them sad, clap, and creepy, etc. To check the movement of audience, we defined the critical point, ${\alpha}$and ${\beta}$. Dynamic State Synchronization was meaningful when the movement value was over critical point ${\beta}$, while Static State Synchronization was effective under critical point ${\alpha}$. ${\beta}$ is made by audience' clapping movement of 10 teams in stead of using average number of movement. After checking the reactive movement of audience, the percentage(%) ratio was calculated from the division of "people having reaction" by "total people". Total 37 teams were made in "2012 Seoul DMC Culture Open" and they involved the experiments. First, they followed induction to clap by staff. Second, basic scene for neutralize emotion of audience. Third, flow scene was displayed to audience. Forth, the reversal scene was introduced. And then 24 teams of them were provided with amuse and creepy scenes. And the other 10 teams were exposed with the sad scene. There were clapping and laughing action of audience on the amuse scene with shaking their head or hid with closing eyes. And also the sad or touching scene made them silent. If the results were over about 80%, the group could be judged as the synchronization and the flow were achieved. As a result, the audience showed similar reactions about similar stimulation at same time and place. Once we get an additional normalization and experiment, we can obtain find the flow factor through the synchronization on a much bigger group and this should be useful for planning contents.

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.