• Title/Summary/Keyword: quantitative PCR

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Conflicting Physiological Characteristics and Aquaporin (JcPIP2) Expression of Jatropha (Jatropha curcas L.) as a Bio-energy Crop under Salt and Drought Stresses (바이오에너지 작물 소재로서 자트로파의 염과 가뭄 스트레스 하에서 상반되는 생리적 특성과 아쿠아포린(JcPIP2)의 발현)

  • Jang, Ha-Young;Lee, Ji-Eun;Jang, Young-Seok;Ahn, Sung-Ju
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.3
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    • pp.183-191
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    • 2011
  • This study was undertaken to collect basic knowledge of Jatropha which is one of bio-energy crops, based on the understanding of physiological and molecular aspects under salt and drought conditions. The treatments were followed as: 100, 200 and 300 mM NaCl for salt stress and 5, 10, 20 and 30% PEG for drought stress for 8 days, respectively. Leaf growth, stomatal conductance, chlorophyll fluorescence and gene expression of aquaporin (JcPIP2) of Jatropha were investigated. From 2 days after treatments, plants treated with higher than 100 mM NaCl and 10% PEG respectively were significantly suppressed in leaf length, width, and stomatal conductance, but 5% PEG treatment showed that plant growth was improved more than control plant. Semi-quantitative RT-PCR analyses revealed that the JcPIP2 gene was expressed in root, stem, cotyledon and leaves. It was not detected in leaves at 200 and 300 mM NaCl treatments. However, transcripts of JcPIP2 were induced in roots and stems under salt and drought conditions compared to those of healthy plants. Therefore, it was concluded that JcPIP2 plays an important role in improving drought tolerance.

Apoptosis and Apoptosis Related Gene Expression in Preimplantation Porcine Diploid Parthenotes Developing In Vitro (착상전 이배체 단위발생 돼지난자의 체외 배양에서 세포사멸과 세포사멸에 관여하는 유전자의 발현에 관한 연구)

  • X. S. Cui;Kim, I. H.;Kim, N. H.
    • Korean Journal of Animal Reproduction
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    • v.27 no.2
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    • pp.169-177
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    • 2003
  • This study was conducted to determine effects of polyvinyl alcohol (PVA), fetal bovine serum (FBS) bovine serum albumin (BSA) and epidermal growth factor (EGF) on blastocoel formation, total cell number, apoptosis and apoptosis-related gene expression of porcine diploid parthenotes developing in vitro. The addition of 0.4% BSA to the culture medium enhanced the development of 2-cell stage parthenotes to the blastocysts stage (P<0.01). FBS reduced cell numbers of blastocysts (P<0.01) and increased percentage of apoptosis in the blastocysts (P<0.001). However, while BSA increased cell numbers, it did so only when EGF was present. Either agent on its own had no effect. Similarly, apoptosis in the blastocysts was not influenced by either agent on its own but was reduced when both BSA and EGF were present. Furthermore, semi-quantitative reverse-transcriptase polymerase chain reaction revealed that EGF enhanced the mRNA expression of Bcl-xL in the presence of 0.4% BSA but BSA and EGF alone had no effect, and EGF and/or BSA did not influence Bak gene expression in the blastocyst stage parthenotes. However FBS reduced Bcl-xL mRNA expression (P <0.05) and enhanced Bak expression. This result suggests that apoptosis related genes expression is significantly affected by supplements, which may result in alteration of apoptosis and embryo viability of porcine embryos developing in vitro.

Trends in Rapid Detection Methods for Food-borne pathogenic Microorganisms by Using New Technologies (신기술 이용 식중독균 신속검출법 개발 동향 분석)

  • Kim, Hyun-Joo;Kim, Yong-Soo;Chung, Myung-Sub;Oh, Deog-Hwan;Chun, Hyang-Sook;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.25 no.4
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    • pp.376-387
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    • 2010
  • Recently, speedy, convenient and easy detection technologies have been developed rapidly and on the contrary, studies on development of traditional detectors applying biochemical characteristics has gradually been decreased. This review examined trend in current studies on detection of food-borne pathogenic microorganisms in the fields of selective media, immuno-assay, Polymerase Chain Reaction (PCR), microarray, terahertz spectroscopy & imagination and so on. Most traditional methods to detect the organisms from food matrix rely on selective media and such a method have disadvantages like long time requirement and distinguishing one species only from each selective medium although they are highly economical. Various new convenient methods such as Enzyme Linked Immuno-sorbent Assay (ELISA), paper-strip kit, fluoroimmunoassay etc. have been developed. The most ideal method for detecting food-borne pathogenic microorganisms in foods should be accurate, convenient, rapid and economical. Additionally, it is needed that capabilities of quantitative analysis and automation to be applied to industries.

The Action Mechanism of Tamoxifen Via Estrogen Receptor on Uterine Leimyoma (자궁근종에서 타목시펜의 수용체를 통한 기전)

  • Lee, Byung-Seok;Cha, Dong-Hyun;Jung, Kyung-Ah;Lee, Hye-Dae;Park, Ki-Hyun;Cho, Dong-Jae;Song, Chan-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.4
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    • pp.337-343
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    • 2002
  • Objectives: To investigate the distribution of $ER{\alpha}$, $ER{\beta}$, c-fos and c-jun in the uterine myoma and myometrium in oder to know how the tamoxifen cause the growth of myoma. Methods: Myoma and myometrial tissue were obtained from the postmenopausal women treated with tamoxifen in the patients with breast cancer and in the premenopausal patients, who were undergoing myoma of uterus from 1998 through 2000. The espression of each gene was quantitated with quantitative RT-PCR. Results: The expression of $ER{\alpha}$ was slightly increased in the myoma than the myometrium in the proliferative phase, and was slightly decreased in the myometrium than the myoma in the secretory phase. However it was not significant statistically. In the postmemopausal women treated with tamoxifen, $ER{\alpha}$ was expressed in all myoma and myome1rial tissues and the expression was not statistically significant. The expression ofER~ was slightly increased in the myome1rium than the leiomyoma in the proliferative and secretory phase, but it was not significant statistically. In the postmemopausal women treated with tamoxifen, the expression of ER~ was significantly incresed in the myome1rium than the leiomyoma. The expression of c-fos was significantly increased in the myome1rium than the leiomyoma in the proliferative and secretory phase. In the postmemopausal women treated with tamoxifen, the expression of c-fos was slightly increased in the leiomyoma than the myomelrium, however, it was not statistically significant. Conclusion: Tamoxifen may cause the growth of leiomyoma by $ER{\alpha}$ with AP-l pathway reducing the counteraction of 6$ER{\beta}$ to $ER{\alpha}$.

Quality characteristics of buckwheat Soksungjang manufactured by Bacillus subtilis HJ18-4 (Bacillus subtilis HJ18-4를 이용하여 제조한 메밀 속성장의 품질특성)

  • Park, Na Young;Lee, Sun Young;Kim, Ji Yeun;Choi, Hye Sun
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.699-704
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    • 2013
  • Buckwheat Soksungjang (BS) is a bealmijang manufactured with buckwheat and soybeans. We manufactured BS using Bacillus subtilis HJ18-4 (HJ18-4), which has high enzyme activities and antibacterial effects. HJ18-4 was inoculated in a different process during the BS manufacturing, which was the meju-making time (Treat 1), and the salt water time was added (Treat 2). The physiochemical and microbial characteristics of the BS were analyzed. As a result, the total aerobic counts (7~8 log CFU/mL) in the BS increased after 15 days of fermentation. Especially, Treat 1 showed higher total aerobic counts and amino-type nitrogen (65.38~202.52 mg%) than Treat 2. During the BS fermentation, the reduction of the sugar contents and the enzyme (protease and amylase) activities decreased. In the relative quantitative expression level of PlcR, Treat 1 did not show toxin gene expressions at the end of the fermentation on Day 23. Treat 1 showed suitable B. cereus physiochemical quality characteristics and inhibition effects. When the modified-form type of fermented soybean paste was manufactured with a single starter, it could not reproduce the natural fermentation quality. These results suggest that the addition of a starter (HJ18-4) in the Meju manufacturing process could enhance the quality characteristics of the manufactured BS via natural fermentation and by suppressing B. cereus.

Marker Assisted Selection-Applications and Evaluation for Commercial Poultry Breeding

  • Sodhi, Simrinder Singh;Jeong, Dong Kee;Sharma, Neelesh;Lee, Jun Heon;Kim, Jeong Hyun;Kim, Sung Hoon;Kim, Sung Woo;Oh, Sung Jong
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.223-234
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    • 2013
  • Poultry industry is abounding day by day as it engrosses less cost of investment per bird as compared to large animals. Poultry have the most copious genomic tool box amongst domestic animals for the detection of quantitative trait loci (QTL) and marker assisted selection (MAS). Use of multiple markers and least square techniques for mapping of QTL affecting quality and production traits in poultry is in vogue. Examples of genetic tests that are available to or used in industry programs are documented and classified into causative mutations (direct markers), linked markers in population-wide linkage disequilibrium (LD) with the QTL (LD markers), and linked markers in population wide equilibrium with the QTL (LE markers). Development of genome-wide SNP assays, role of 42 K, 60 K (Illumina) and 600 K (Affymetrix$^{(R)}$ Axim$^{(R)}$) SNP chip with next generation sequencing for identification of single nucleotide polymorphism (SNP) has been documented. Hybridization based, PCR based, DNA chip and sequencing based are the major segments of DNA markers which help in conducting of MAS in poultry. Economic index-marker assisted selection (EI-MAS) provides platform for simultaneous selection for production traits while giving due weightage to their marginal economic values by calculating predicted breeding value, using information on DNA markers which are normally associated with relevant QTL. Understanding of linkage equilibrium, linkage dis-equilibrium, relation between the markers and gene of interest are quite important for success of MAS. This kind of selection is the most useful tool in enhancing disease resistance by identifying candidate genes to improve the immune response. The application of marker assisted selection in selection procedures would help in improvement of economic traits in poultry.

Calmodulin of Olive Flounder Paralichthys olivaceus : Cloning and Expression Analysis

  • Hong, Gyeong-Eun;Kong, Hee Jeong;Nam, Bo-Hye;Kim, Young-Ok;Kim, Woo-Jin;Lee, Sang-Jun;Choi, Tae-Jin
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.4
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    • pp.234-237
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    • 2007
  • Calmodulin (CaM) is a $Ca^{2+}$-binding protein essential for biological functions mediated through $Ca^{2+}$-dependent mechanism. A cDNA clone for CaM was isolated from a cDNA library of olive flounder Paralichthys olivaceus. The CaM cDNA concists of 782 bp and encodes a polypeptide of 149 amino acids with four $Ca^{2+}$-binding motifs EF-hands (EF-I, EF-II, EF-III, and EF-IV). The deduced amino acid sequence of CaM shows 97-100% amino acid sequence identity to other CaM sequences. Semi-quantitative PCR analysis revealed that the CaM transcription was began during early development and the CaM mRNA is expressed highly in brain and intestine, and moderately in kidney, gill, and eye of healthy olive flounder. Taken together, CaM may be necessary for early olive flounder development and that it may have a part in homeostasis.

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MMP and TIMP production in periodontal ligament fibroblasts stimulated by Prevotella nigrescens lipopolysaccharide (Prevotella nigrescens lipopolysaccharide로 자극한 치주인대 섬유아세포에서 기질금속단백분해효소와 단백분해효소억제제의 생성 양상에 대한 연구)

  • Yang, Won-Kyung;Lee, Woo-Cheol;Kim, Mi-Ri;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.30 no.5
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    • pp.372-384
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    • 2005
  • The purpose of this study was to monitor the secretion of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) by human periodontal ligament (PDL) fibroblasts stimulated with Prevotella nigrescens lipopolysaccharide (LPS), and to examine the effect of calcium hydroxide treatment on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. PDL fibroblasts were stimulated by the LPS (0, 0.1, 1, 10 ${\mu}g/ml$) or LPS (10 ${\mu}g/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days, for various periods of time (12, 24, 48 h). Immunoprecipitation were performed for protein level analysis of MMP-1 MMP-2 and TIMP-1. Total RNA was isolated and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 mRNA. According to this study, the results were as follows: 1. The p개duction of MMP-1 by stimulation with P. nigrescens LPS increased in time-dependent manner, and showed maximum value at 48 h in both protein and mRNA level. But there was no dose-dependent increas. 2. MMP-2 production time-dependently increased when stimulated with 1 and 10 ${\mu}g/ml$LPS, but there was no dose-dependent increase. 3. TIMP-1 p개duction increased to 24 h, but decreased at 48 h. It increased when stimulated with 0.1 and 1${\mu}g/ml$, but suppressed at 10 ${\mu}g/ml$ .4. P. nigrescens LPS pretreated with $Ca(OH)_2$ markedly downregulated MMP-1 gene expression.

Effect of microgrooves and fibronectin conjugation on the osteoblast marker gene expression and differentiation

  • Park, Su-Jung;Leesungbok, Richard;Ahn, Su-Jin;Im, Byung-Jin;Lee, Do Yun;Jee, Yu-Jin;Yoon, Joon-Ho;Cui, Taixing;Lee, Sang Cheon;Lee, Suk Won
    • The Journal of Advanced Prosthodontics
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    • v.7 no.6
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    • pp.496-505
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    • 2015
  • PURPOSE. To determine the effect of fibronectin (FN)-conjugated, microgrooved titanium (Ti) on osteoblast differentiation and gene expression in human bone marrow-derived mesenchymal stem cells (MSCs). MATERIALS AND METHODS. Photolithography was used to fabricate the microgrooved Ti, and amine functionalization (silanization) was used to immobilize fibronectin on the titanium surfaces. Osteoblast differentiation and osteoblast marker gene expression were analyzed by means of alkaline phosphatase activity assay, extracellular calcium deposition assay, and quantitative real-time PCR. RESULTS. The conjugation of fibronectin on Ti significantly increased osteoblast differentiation in MSCs compared with non-conjugated Ti substrates. On the extracellular calcium deposition assays of MSCs at 21 days, an approximately two-fold increase in calcium concentration was observed on the etched 60-${\mu}m$-wide/10-${\mu}m$-deep microgrooved surface with fibronectin (E60/10FN) compared with the same surface without fibronectin (E60/10), and a more than four-fold increase in calcium concentration was observed on E60/10FN compared with the non-etched control (NE0) and etched control (E0) surfaces. Through a series of analyses to determine the expression of osteoblast marker genes, a significant increase in all the marker genes except type I collagen ${\alpha}1$ mRNA was seen with E60/10FN more than with any of the other groups, as compared with NE0. CONCLUSION. The FN-conjugated, microgrooved Ti substrate can provide an effective surface to promote osteoblast differentiation and osteoblast marker gene expression in MSCs.

Overexpression of MicroRNA-31 as a Promising Biomarker for Prognosis and Metastasis in Human Colorectal Cancer (MicroRNA-31 과발현을 이용한 대장암의 예후예측 및 전이예측 바이오마커 발굴)

  • Hur, Keun
    • Journal of Life Science
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    • v.26 no.6
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    • pp.705-710
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    • 2016
  • Colorectal cancer (CRC) is the third most common cancer and a leading cause of cancer-related death worldwide. Although several diagnostic and therapeutic tools have been available, CRC remains difficult to complete cure because of insufficient understanding of the molecular mechanisms underlying this disease progression. MicroRNAs (miRNAs) are small non-coding RNA molecules that strongly regulate gene expression via transcriptional and translational control mechanisms. Many crucial cellular pathways are frequently disrupted in cancer development process due to dysregulation of several miRNAs. Mir-31 functions as an oncogene that modulate expression of multiple cancer related genes. Thus, we aimed to demonstrate clinical relevance of miR-31 in human CRC. Quantitative RT-PCR analysis of miR-31 expression was performed in 175 CRC tissues and 16 normal colonic mucosa (NM). Next, we investigated clinical significances of miR-31 expression in various clinicopathologic features in CRC patients cohort. Mir-31 was significantly up-regulated in CRC tissues compared to NM. In CRC tissues, miR-31 expression level was significantly elevated in a stage-dependent manner, which was associated with poor survival in patients with CRC. High miR-31 levels in CRC tissues significantly correlated with poor prognosis (hazard ratio [HR]=2.4) as well as distant metastasis (odds ratio [OR]=2.3). In conclusion, we identified clinical significance of miR-31 expression in CRC. High miR-31 expression may be clinically able to use as a biomarker for CRC prognosis and predicting metastasis.