• Title/Summary/Keyword: quantitative expression analysis

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Screening for MiRNAs Related to Laryngeal Squamous Carcinoma Stem Cell Radiation

  • Huang, Chang-Xin;Zhu, Ying;Duan, Guang-Liang;Yao, Ji-Fen;Li, Zhao-Yang;Li, Da;Wang, Qing-Qing
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4533-4537
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    • 2013
  • Objective: To use microarray chip technology for screening of stem cell radiation related miRNAs in laryngeal squamous cell carcinoma; study and explore the relationship of miRNAs with radiosensitivity of laryngeal squamous cells. Method: After conventional culture and amplification of the laryngeal squamous carcinoma cell line Hep-2, CD 133+ cells were screened out with combination of isolated culture of stem cell microspheres and FACS for preparation of laryngeal cancer stem cells. After radiation treatment, miRNAs of laryngeal squamous carcinoma stem cells before and after radiation were enriched and purified. After microarray hybridization with mammalian miRNA and scanning of fluorescence signal, the miRNAs of laryngeal squamous carcinoma stem cells before and after radiation was subject to differential screening and clustering analysis. Real-time quantitative RT-PCR was used to verify part of the differentially expressed miRNAs. Results: 70 miRNAs related to laryngeal cancer stem cell radiation with 2-fold difference in expression were screened out, in which 62 were down-regulated and 8 were up-regulated. Fluorescent quantitative RT-PCR results were consistent with miRNAs chip results. Conclusion: Some miRNAs may be involved in self-regulation with laryngeal squamous carcinoma stem cell radiation.

Molecular Cloning, Protein Expression, and Regulatory Mechanisms of the Chitinase Gene from Spodoptera littoralis Nucleopolyhedrovirus

  • Yasser, Norhan;Salem, Reda;Alkhazindar, Maha;Abdelhamid, Ismail A.;Ghozlan, Said A.S.;Elmenofy, Wael
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.305-315
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    • 2021
  • The cotton leafworm, Spodoptera littoralis, is a major pest in Egypt and many countries worldwide, and causes heavy economic losses. As a result, management measures to control the spread of the worm are required. S. littoralis nucleopolyhedrovirus (SpliNPV) is one of the most promising bioagents for the efficient control of insect pests. In this study, a chitinase gene (chitA) of a 1.8 kb DNA fragment was cloned and fully characterized from SpliNPV-EG1, an Egyptian isolate. A sequence of 601 amino acids was deduced when the gene was completely sequenced with a predicted molecular mass of 67 kDa for the preprotein. Transcriptional analyses using reverse transcription polymerase chain reaction (RT-PCR) revealed that chitA transcripts were detected first at 12 h post infection (hpi) and remained detectable until 168 hpi, suggesting their transcriptional regulation from a putative late promoter motif. In addition, quantitative analysis using quantitative RT-PCR showed a steady increase of 7.86-fold at 12 hpi in chitA transcription levels, which increased up to 71.4-fold at 120 hpi. An approximately 50 kDa protein fragment with chitinolytic activity was purified from ChitA-induced bacterial culture and detected by western blotting with an anti-recombinant SpliNPV chitinase antibody. Moreover, purification of the expressed ChitA recombinant protein showed in vitro growth inhibition of two different fungi species, Fusarium solani and F. oxysporum, confirming that the enzyme assembly and activity was correct. The results supported the potential role and application of the SpliNPV-ChitA protein as a synergistic agent in agricultural fungal and pest control programs.

Expression of Endometriosis Related Genes in the Shed Endometrial Tissues from Menstrual Blood (생리혈에 존재하는 자궁내막조직에서 자궁내막증 관련 유전자의 발현 양상)

  • Park, Chan-Woo;Jun, Jin-Hyun;Koong, Mi-Kyoung;Song, In-Ok
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.4
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    • pp.275-283
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    • 2007
  • Objective: This study was performed to investigate the expressions of endometriosis related genes in shed endometrial tissues from menstrual blood of patients with or without endometriosis. Methods: The shed endometrial tissues were collected on 2$^{nd}$ or 3$^{rd}$ day of menstrual cycle with Wallace catheter in patients with endometriosis (n=16) and without endometriosis (n=26). The mRNA expressions of twelve kinds of endometriosis related genes were compared between two groups using semi-quantitative RT-PCR. Results: The collected shed endometrium was confirmed by histological observation. Expressions of telomerase, c-kit and aromatase mRNA were not detected by RT-PCR in shed endometrial tissues. The mRNA expressions of apoptosis related genes (fas, fas ligand, bcl-2, bax), stem cell factor, estrogen receptor-$\alpha$/$\alpha$, endometriosis protein-I and secretory leukocyte protease inhibitor gene were similar between shed endometrial tissues with endometriosis and without endometriosis. Conclusion: We could not find the difference of mRNA expressions of tested endometriosis related genes between shed endometrial tissues with or without endometriosis by semi-quantitative RT-PCR analysis. It may be related to the dynamical changes of gene expressions in the endometrium with menstrual cycle.

Molecular Cloning and Expression Analysis of Red-spotted Grouper, Epinephelus akaara Hsp70 (수온변화에 따른 붉바리(Epinephelus akaara)의 heat shock protein (Hsp) 70 mRNA 발현)

  • Min, Byung Hwa;Hur, Jun Wook;Park, Hyung Jun
    • Journal of Life Science
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    • v.28 no.6
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    • pp.639-647
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    • 2018
  • A new heat shock protein 70 was identified in red-spotted grouper (Epinephelus akaara) based on an expression analysis. The cDNA of red-spotted grouper Hsp70 (designated RgHsp70) was cloned by the rapid amplification of cDNA ends (RACE) techniques. The full-length of RgHsp70 cDNA was 2,152 bp, consisting of a 5'-terminal untranslated region (UTR) of 105 bp, a 3'-terminal UTR of 274 bp, and an open reading frame (ORF) of 1,773 bp that encode a polypeptide of 590 amino acids with a theoretical molecular weight of 64.9 kDa and an estimated isoelectric point of 5.2. Multiple alignment and phylogenetic analyses revealed that the RgHsp70 gene shares a high similarity with other Hsp70 fish genes. RgHsp70 contained all three classical Hsp70 family signatures. The results indicated the RgHsp70 is a member of the heat shock protein 70 family. RgHsp70 mRNA was predominately expressed in the liver, with reduced expression noted in the head-kidney tissues. The expression analysis of different water temperatures (21, 18, 15 and $12^{\circ}C$) for sampled livers revealed that expression gradually increased at $12^{\circ}C$ compared to $21^{\circ}C$. In this study, the effects of water temperature lowering on the physiological conditions were investigated, and the results revealed that novel RgHsp70 may be an important molecule involved in stress responses.

Nutriproteomics: Identifying the Molecular Targets of Nutritive and Non-nutritive Components of the Diet

  • Barnes, Stephen;Kim, Helen
    • BMB Reports
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    • v.37 no.1
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    • pp.59-74
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    • 2004
  • The study of whole patterns of changes in protein expression and their modifications, or proteomics, presents both technological advances as well as formidable challenges to biological researchers. Nutrition research and the food sciences in general will be strongly influenced by the new knowledge generated by the proteomics approach. This review examines the different aspects of proteomics technologies, while emphasizing the value of consideration of "traditional" aspects of protein separation. These include the choice of the cell, the subcellular fraction, and the isolation and purification of the relevant protein fraction (if known) by protein chromatographic procedures. Qualitative and quantitative analyses of proteins and their peptides formed by proteolytic hydrolysis have been substantially enhanced by the development of mass spectrometry technologies in combination with nanoscale fluidics analysis. These are described, as are the pros and cons of each method in current use.

Comparative Proteome Analysis of Celastrol-Treated Helicobacter pylori

  • Kim, Sa-Hyun
    • Biomedical Science Letters
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    • v.23 no.4
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    • pp.395-401
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    • 2017
  • Various preclinical and clinical trials have been conducted the efficacy of celastrol. In data presented in the current manuscript is the first trial to inhibit Helicobacter pylori with celastrol. In this study, the quantitative change of various H. pylori proteins including CagA and VacA by the anti-bacterial effect of celastrol was determined. The anti-H. pylori effects of celastrol was investigated by performing 2-dimensional electrophoresis and additional supporting experiments. After 2-dimensional electrophoresis analysis, spot intensities were analyzed and then each spot was identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) or peptide sequencing using Finnigan LCQ ion trap mass spectrometer (LC-MS/MS). The results show that celastrol has multiple effects on protein expression in H. pylori.

Analysis the static thrust force and dynamic thrust force in HB-type Linear Pulse Motor (하이브리드형 선형 펄스모터의 정추력 및 동추력 분석)

  • Kim, Dong-Hee;Ahn, Jae-Young;Kim, Kwang-Heon
    • Proceedings of the KIPE Conference
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    • 2007.07a
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    • pp.214-216
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    • 2007
  • The linear motor is available for linear transition motion, because of its advantages, the motor design and its application have gradually increased, but the quantitative measurement system of thrust force has not been generalized. Need analysis of correct thrust for control performance improvement of HB-LPM(HB-type Linear Pulse Motor). It is difficult to analyze HB-LPM's thrust. In this paper, HB-LPM's thrust is expressed to mathematical expression. And it is proved validity of this numerical formula by thrust measurement system. Two phase driver is composed. It is verified validity of numerical formula that measure waveform of electric current and voltage that is supplied in each phase.

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Development of Diagnosis of Trouble Model for Effective Operation of Air-compressor (효율적인 공기압축기 운영을 위한 이상진단모델 연구)

  • Im, Sang Don;Jung, Young Deuk;Kim, Jong Rae
    • Journal of the Korea Safety Management & Science
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    • v.16 no.3
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    • pp.239-248
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    • 2014
  • Most systems used in industrial sites, actually have non-linearity and uncertainty. Therefore there are a lot of difficulties in evaluating conditions of these systems. Generally, the quantitative analysis and expression are found hard because the general public cannot easily make an accurate interpretation on the systems. Thus development of a system that utilizes an expertise from skilled analysts is required. In this research, a real-time sensor signal conditioning system and Fuzzy-expert system have been separately set up into an inference algorithm. So that it ensures a fast, accurate, objective and quantitative operational condition value provided to the manager. Therefore, FE_AFCDM is suggested in this literature, as an effective system for diagnosing the problems related to the air compressor. It can quantify the uncertain and absurd condition to operate the air compressor facilities safely and financially.

Effects of Five Fractions of Artemisia capillaris THUNB on $TGF{\beta}1-induced$ Apoptosis in HepG2 Cells (인진분획물이 인체간세포의 $TGF{\beta}1-induced$ Apoptosis에 미치는 영향)

  • 이지현;이장훈;우홍정
    • The Journal of Korean Medicine
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    • v.21 no.1
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    • pp.53-61
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    • 2000
  • Objectives: This study was camed out to examine the effect of five fractions of aqueous extract from Artemisia capillaris THUNB(ACT), on TGF, 1-induced apoptosis, cell viability, cell cycle progression and mRNA expression of apoptosis-related genes in human hepatocyte cell line HepG2. Methods: This study employed Tryphan blue exclusion assay, DNA fragmentation assay, Cpp32 protease activity assay and Quantitative RT-PCR analysis. Results: In the Tryphan blue exclusion assay, the butanol fraction of ACT with $TGF{\beta}$, l showed magnificent (Nice word, ut is it appropriate in a medical abstract\ulcorner) viability and the H2O fraction of ACT with $TGF{\beta}$, l also showed higher viability than only $TGF{\beta}$, l-treated group. DNA fragmentation assay showed that the butanol fraction and the H2O fraction carried inhibitory effects on apoptosis induction, with the butanol fraction displaying greater effects. The Cpp32 protease activity assay showed that the butanol fraction decreased Cpp32 protease activity. The H2O fraction of ACT had no significant effect on the Cpp32 protease activity. Quantitative RT-PCR showed that the butanol fraction suppressed Bax, p 15/INK4B, p21/Waf1, PAI-1 and increased Bcl-2 gene. Conclusions: The data shows that butanol fraction of ACT increases the hepatocyte viability and has the hepatocellular protective effect by the suppression of $TGF{\beta}$, l induced-apoptosis through gene regulation.

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Characterization of Quantitative Trait Loci (QTL) for Growth using Genome Scanning in Korean Native Pig

  • Lee, H.K.;Choi, I.S.;Choi, B.H.;Kim, T.H.;Jung, I.J.
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.107-112
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    • 2004
  • Molecular genetic markers were genotyped used to detect chromosomal regions which contain economically important traits such as growth traits in pigs. Three generation resource population was constructed from a cross between the Korean native boars and Landrace sows. A total of 193 F2 animals from intercross of F1 were produced. Phenotypic data on 7 traits, birth weight, body weight at 3, 5, 12, 30 weeks of age, live empty weight were collected for F2 animals. Animals including grandparents (F0), parents (F1), offspring (F2) were genotyped for 194 microsatellite markers covering from chromosome 1 to 18. Quantitative trait locus analyses were performed using interval mapping by regression under line-cross model. To characterize presence of imprinting, genetic full model in which dominance, additive and imprinting effect were included was fitted in this analysis. Significance thresholds were determined by permutation test. Using imprinting full model, four QTL with expression of imprinted effect were detected at 5% chromosome-wide significance level for growth traits on chromosome 1, 5, 7, 13, 14, and 16.