• Title/Summary/Keyword: reverse transcriptase polymerase chain reaction

Search Result 349, Processing Time 0.033 seconds

Characteristics of outbreak for the classical swine fever (CSF) at Incheon metropolitan area in 2002 (인천지역에서 발생한 돼지콜레라의 역학적 특성)

  • 권효정;변재원;이정구;김경호;박은정;이성모;황현순
    • Korean Journal of Veterinary Service
    • /
    • v.26 no.1
    • /
    • pp.1-9
    • /
    • 2003
  • The aim of this survey was to investigate the characteristics of outbreak farm determined as the classical swine fever(CSF) at Gangwha-gun and Seo-gu, Incheon metropolitan area from October 7 to November 25 in 2002. Sixty pigs in six different farms were confirmed to the CSF and a total of 9,106 pigs containing 3,194 related epidemiologically was slaughtered to stop spreading of the disease. Clinical signs of pigs diagnosed with the CSF were high fever, anorexia, depression, paralysis of hindlimbs, cyanosis, etc and gross lesions were typically represented with hemorrhage of submandibular and superficial lymph node, infarction of spleen, and petechial (ecchymotic) hemorrhage of kidney and skin. But some outbreak farms had not shown remarkable symptoms, so they were confused with other bacterial diseases. White blood cell (WBC) counts, the classical swine fever virus(CSFV) antigen and antibody enzyme linked immunosorbent assay(ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR) results about six farms indicated that total 60 pigs were infected with the CSFV. Although the origin and infection route of the CSFV were not clear, but the transmissions between farms were mainly through indirect contact such as the movement of farm personal and vehicles from outbreak farm.

Analysis of gene expression during odontogenic differentiation of cultured human dental pulp cells

  • Seo, Min-Seock;Hwang, Kyung-Gyun;Kim, Hyong-Bum;Baek, Seung-Ho
    • Restorative Dentistry and Endodontics
    • /
    • v.37 no.3
    • /
    • pp.142-148
    • /
    • 2012
  • Objectives: We analyzed gene-expression profiles after 14 day odontogenic induction of human dental pulp cells (DPCs) using a DNA microarray and sought candidate genes possibly associated with mineralization. Materials and Methods: Induced human dental pulp cells were obtained by culturing DPCs in odontogenic induction medium (OM) for 14 day. Cells exposed to normal culture medium were used as controls. Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR). We also performed a gene set enrichment analysis (GSEA) of the microarray data. Results: Six hundred and five genes among the 47,320 probes on the BeadChip differed by a factor of more than two-fold in the induced cells. Of these, 217 genes were upregulated, and 388 were down-regulated. GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated. Conclusions: Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.

Beauvericin, a cyclic peptide, inhibits inflammatory responses in macrophages by inhibiting the NF-κB pathway

  • Yoo, Sulgi;Kim, Mi-Yeon;Cho, Jae Youl
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.21 no.4
    • /
    • pp.449-456
    • /
    • 2017
  • Beauvericin (BEA), a cyclic hexadepsipeptide produced by the fungus Beauveria bassiana, is known to have anti-cancer, anti-inflammatory, and anti-microbial actions. However, how BEA suppresses macrophage-induced inflammatory responses has not been fully elucidated. In this study, we explored the anti-inflammatory properties of BEA and the underlying molecular mechanisms using lipopolysaccharide (LPS)-treated macrophage-like RAW264.7 cells. Levels of nitric oxide (NO), mRNA levels of transcription factors and the inflammatory genes inducible NO synthase (iNOS) and interleukin (IL)-1, and protein levels of activated intracellular signaling molecules were determined by Griess assay, semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), luciferase reporter gene assay, and immunoblotting analysis. BEA dose-dependently blocked the production of NO in LPS-treated RAW264.7 cells without inducing cell cytotoxicity. BEA also prevented LPS-triggered morphological changes. This compound significantly inhibited nuclear translocation of the $NF-{\kappa}B$ subunits p65 and p50. Luciferase reporter gene assays demonstrated that BEA suppresses MyD88-dependent NF-${\kappa}B$ activation. By analyzing upstream signaling events for $NF-{\kappa}B$ activation and overexpressing Src and Syk, these two enzymes were revealed to be targets of BEA. Together, these results suggest that BEA suppresses $NF-{\kappa}B$-dependent inflammatory responses by suppressing both Src and Syk.

Development of Recombinant Saccharomyces cerevisiae Expressing Epoxide Hydrolase for the Preparation of Chiral Phenyl Oxirane (광학활성 Phenyl Oxirane 제조용 유전자 재조합 생촉매 개발)

  • 이수정;이은정;김초희;이지원;김희숙;이은열
    • Journal of Life Science
    • /
    • v.13 no.1
    • /
    • pp.105-109
    • /
    • 2003
  • Epoxide hydrolase (EH) gene from Aspergillus niger #33 was cloned from cDNA library generated by reverse transcriptase-polymerase chain reaction (RT-PCR). The nucleotide sequence analysis revealed that the deduced amino acid sequence was almost similiar to that of A. niger LCP521 previously reported. The cloned EH gene was transformed into Saccharomyces cerevisiae and expressed by addition of galactose. The recombinant S. cerevisiae showed hydrolytic activity toward racemic phenyl oxirane substrate based on chiral GC analysis, and can be used as a potential biocatalyst for the preparation of chiral phenyl oxirane.

Expression of DSPP mRNA During Differentiation of Human Dental Pulp-derived Cells (HDPC) and Transplantation of HDPC Using Alginate Scaffold

  • Aikawa, Fumiko;Nakatsuka, Michiko;Kumabe, Shunji;Jue, Seong-Suk;Hayashi, Hiroyuki;Shin, Je-Won;Iwai, Yasutomo
    • International Journal of Oral Biology
    • /
    • v.31 no.3
    • /
    • pp.73-79
    • /
    • 2006
  • Tissue stem cells are used for the regenerative medicine. In previous study we observed hard tissue formation of human dental pulp-derived cells using alginate scaffold. In this study, we explore the ability to differentiate of the 13th passage cells with glycerol 2-phosphate disodium salt hydrate (${\beta}-GP$) which accelerate calcification. Reverse transcriptase Polymerase Chain Reaction (RT-PCR), transplants using alginate scaffold and histological examination were performed. We observed the expression of DSPP mRNA on day 10 cultured cells with ${\beta}-GP$. In conclusion, the 13th passage cells still have an ability to differentiate into odontoblast-like cells and alginate supports the differentiation of cultured cells in the transplants.

Usefulness of Chimeric Transcript in the Diagnosis of Pediatric Solid Tumors (소아악성고형종의 진단에 있어서 chimeric transcript의 유용성)

  • Choi, Seung-Hoon
    • Advances in pediatric surgery
    • /
    • v.5 no.1
    • /
    • pp.45-52
    • /
    • 1999
  • Pediatric solid tumors have many histologic similarity. These tumors contained small round cell types, and cause frequent diagnostic problems in pediatric pathology. An important advance in the differentiation of these small round cell tumors has been the identification of consistent chromosomal translocations associated with several types of tumors. Eighteen patients with soft tissue sarcoma were available for review. Seventeen cell lines were also included in this study. The RNA from the specimens were analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). PAX3-FKHR fusion was present in four of five alveolar rhabdomyosarcoma and PAX7-FKHR fusion was detected in one of five alveolar rhabdomyosarcoma. None of the specimens expressed more than one chimeric transcript. EWS-FLI1 or EWS-ERG fusions were detected in all seven Ewings' sarcoma. No specimens showed EWS-WT1 fusion. These results corresponded well to the histopathologic diagnosis. There were no differences in the histologic appearances of tumors with the more frequent PAX3-FKHR or EWS-FLI1 fusions compared with those containing the variant PAX7-FKHR or EWS-ERG fusions. RT-PCR assay for chimeric transcript is a useful tool for rapid and objective diagnosis of pediatric solid tumors. Through these tools, we can approach genetically to the differential diagnosis of undifferentiated small round tumors.

  • PDF

Effects of geagibokrounghwan on superoxide dismutase gene expression in experimental mouse fed cholesterol (계지복령환이 Cholesterol 식이 생쥐의 SOD 유전자발현능에 미치는 영향)

  • Hong Yun-Pyo;Ro Sung-Hyun
    • Herbal Formula Science
    • /
    • v.6 no.1
    • /
    • pp.215-226
    • /
    • 1998
  • Geagibokrounghwan (桂技茯笭湯) has long been used to cure human diseases such as vascular and blood disorders. However, it is still unkown on its action mechanism, physiolosical and biochemical meaning. Thus, many attempts were tried to show the scientific background covering the above mentioned mechanism. The effect of Geagibokrounghwan, which was known to date, as follow; effective circulation of body blood system, proliferation of leucocytes and antioxidative action. In this study, we have applied the Geagibokrounghwan administration and feed to mouse, to see effects on the expression of superoxide dismutase(SOD) mRNA as antioxidative agent and oxygen radical scavenger. Total RNAs includingmRNA have been isolated from liver and white blood cells after mice were fed with cholesterol in high dose. Also, in a separate group, the cholesterol-administrated mice were fed with Geagibokrounghwan to see the effects on SOD transcription. and then reverse transcriptase-polymerase chain reaction (RT-PCR) usion each primer set (SOD-F;GATGAAAGCGGTGT-3'; SOD-R; 5'-CCTGTGGAGTGATT-3') were performed to trace theamounts of mRNA. SOD mRNA was specifically expressed in Geagibokrounghwan-fed mice at 2 weeks after treatment, however, gradually reduced after 4 weeks. These results indicate that Geagibokrounghwan is highly applicable in treatment of the above mentioned human diseases.

  • PDF

Detection and genetic characteristic of Yellow-head virus genotype 8 (YHV-8) Cultured Litopanaeus vanamei, in Korea (국내 양식 흰다리새우에서 검출된 노랑머리바이러스(YHV-8)의 유전학적 특성)

  • Kim, Seok-Ryel;CWR, Gunasekara;SHMP, Wimalasena;Shin, Gee-Wook
    • Journal of fish pathology
    • /
    • v.33 no.1
    • /
    • pp.77-81
    • /
    • 2020
  • This study was performed for screening yellow head virus(YHV) complex in 252 including 235 white leg shrimps (Litopenaeus vannamei) and 17 oriental shrimp (Fenneropenaeus chinensis) collected from 18 farms located in southwestern province of Korea. The virus complex was detected by nested reverse-transcriptase polymerase chain reaction (RT-PCR) assay. In the assay, amplicons were resulted in RNAs exracted from 38 shrimps (21 white leg shrimps and 17 oriental shrimps) obtained from 7 farms. In phylogenetic analysis using sequences of ORF1b gene, all 38 sequences obtained in this study formed an independent lineage with YHV-8 genotype firstly isolated in China, belonged to an YHV-8 clade.

Arsenic-Induced Differentially Expressed Genes Identified in Medicago sativa L. roots

  • Rahman, Md. Atikur;Lee, Sang-Hoon;Kim, Ki-Yong;Park, Hyung Soo;Hwang, Tae Young;Choi, Gi Jun;Lee, Ki-Won
    • Journal of The Korean Society of Grassland and Forage Science
    • /
    • v.36 no.3
    • /
    • pp.243-247
    • /
    • 2016
  • Arsenic (As) is a toxic element that easily taken up by plants root. Several toxic forms of As disrupt plant metabolism by a series of cellular alterations. In this study, we applied annealing control primer (ACP)-based reverse transcriptase PCR (polymerase chain reaction) technique to identify differentially expressed genes (DEGs) in alfalfa roots in response to As stress. Two-week-old alfalfa seedlings were exposed to As treatment for 6 hours. DEGs were screened from As treated samples using the ACP-based technique. A total of six DEGs including heat shock protein, HSP 23, plastocyanin-like domain protein162, thioredoxin H-type 1 protein, protein MKS1, and NAD(P)H dehydrogenase B2 were identified in alfalfa roots under As stress. These genes have putative functions in abiotic stress homeostasis, antioxidant activity, and plant defense. These identified genes would be useful to increase As tolerance in alfalfa plants.

O-Methyltransferases from Arabidopsis thaliana

  • Kim, Bong-Gyu;Kim, Dae-Hwan;Hur, Hor-Gil;Lim, Jun;Lim, Yoong-Ho;Ahn, Joong-Hoon
    • Journal of Applied Biological Chemistry
    • /
    • v.48 no.3
    • /
    • pp.113-119
    • /
    • 2005
  • O-methylation mediated by O-methyltransferases (OMTs) is a common modification in natural product biosynthesis and contributes to diversity of secondary metabolites. OMTs use phenylpropanoids, flavonoids, other phenolics and alkaloids as substrates, and share common domains for S-adenosyl-L-methionine (AdoMet) and substrate binding. We searched Arabiposis genome and found 17 OMTs genes (AtOMTs). AdoMet- and substrate-binding sites were predicted. AdoMet binding domain of AtOMTs is highly conserved, while substrate-binding domain is diverse, indicating use of different substrates. In addition, expressions of six AtOMT genes in response to UV and in different tissues were investigated using real-time quantitative reverse transcriptase-polymerase chain reaction. All the AtOMTs investigated were expressed under normal growth condition and most, except AtOMT10, were induced after UV illumination. AtOMT1 and AtOMT8 were expressed in all the tissues, whereas AtOMT10 showed flower-specific expression. Analysis of these AtOMT gene expressions could provide some clues on AtOMT involvement in the cellular processes.