• 제목/요약/키워드: Reverse transcriptase polymerase chain reaction

검색결과 349건 처리시간 0.026초

Effects of bacterial LPS and DNA on the induction of IL-1β, IL-10 and IL-12 by mouse peritoneal macrophages in vitro

  • Samad, D. Abdel;Abdelnoor, AM
    • Advances in Traditional Medicine
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    • 제6권2호
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    • pp.134-143
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    • 2006
  • The capacities of bacterial DNA, extracted from Salmonella typhimurium, and lipopolysaccharide (LPS), extracted from Salmonella minnesota, to activate mouse peritoneal macrophages in vitro were compared. Activation was assessed by estimating e levels of 3 cytokines, IL-10, IL-12, and $IL-1{\beta}$, at time intervals of 3, 6, 9, and 24 h after addition of LPS and/or DNA to macrophage cultures. Cytokine levels in culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA) and cytokine mRNA levels were estimated based on band intensity in cultured cells by reverse transcriptase-polymerase chain reaction (RT-PCR). Results obtained demonstrated the ability of DNA and LPS to elicit increased production of all 3 cytokines as compared to controls. In the amount tested, LPS appeared to be a more potent inducer of IL-12, and $IL-1{\beta}$, whereas DNA induced higher levels of IL-10. DNA and LPS, used in combination, exhibited neither an additive nor a synergistic effect. Rather, an antagonist effect appeared to occur. RT-PCR results correlated well with ELISA.

Expression of CD320 in human B cells in addition to follicular dendritic cells

  • Cho, Wha-Jung;Choi, Jin-Suk;Park, Chan-Hum;Yoon, Sun-Ok;Jeoung, Doo-Il;Kim, Young-Myeong;Choe, Jong-Seon
    • BMB Reports
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    • 제41권12호
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    • pp.863-867
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    • 2008
  • CD320 has been recently discovered and reported as a follicular dendritic cell (FDC) protein. Although CD320 is known to enhance proliferation of germinal center (GC) B cells, little other information is available. In this study, we investigated its cellular distribution in the GC. Confocal microscopy of human tonsil sections revealed co-localization of CD320 with CD19 and CD38 but not with CD3 indicating that GC B cells expressed CD320 in addition to FDC. In purified GC B cells, CD320 expression was inhibited in the nucleus, membrane and cytoplasm. Reverse transcriptase-polymerase chain reaction confirmed CD320 mRNA expression in B cells. These finding indicate that CD320 is expressed in B cells in addition to FDC, and that its GC activity may be more complicated than previously thought.

Isolation and Characterization of Terpene Synthase Gene from Panax ginseng

  • Kim, Yu-Jin;Ham, Ah-Rom;Shim, Ju-Sun;Lee, Jung-Hye;Jung, Dae-Young;In, Jun-Gyo;Lee, Bum-Soo;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • 제32권2호
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    • pp.114-119
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    • 2008
  • Terpene synthase plays a key role in biosynthesis of triterpene saponins (ginsenosides) and is intermediate in the biosynthesis of a number of secondary metabolites. A terpene synthase (PgTPS) cDNA was isolated and characterized from the root of Panax ginseng c.A. Meyer. The deduced amino acid sequence of PgTPS showed a similarity with A. deliciosa (AAX16121) 61%, V. vinifera (AAS66357) 61%, L. hirsutum (AAG41891) 55%, M. truncatula (AAV36464) 52%. And the segment of a terpene synthase gene was amplified by reverse transcriptase-polymerase chain reaction (RTPCR). We studied expression of terpene synthase under stressful conditions like chilling, salt, UV, and heavy metal stress treatment. Expression of PgTPS was increased gradually after exposure to stresses such as chilling, salt, and UV illumination. But its transcription seems to be reduced by cadmium and copper treatment.

오레가노 추출물의 모발성장 촉진효과 (Effect of Origanum vulgare Extracts on Hair Regeneration)

  • 박장순
    • 생약학회지
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    • 제44권3호
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    • pp.275-280
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    • 2013
  • This study was carried out to investigate the effect of Origanum vulgare extracts on cell proliferation of human hair dermal papilla cell (HHDPC) using sulforhodamine B (SRB) assay, antioxidant activity by 1,1-diphenyl-2-picryl hydrazyl (DPPH) method, expression of insulin-like growth factor-1 (IGF-1) by analyzing reverse transcriptase polymerase chain reaction (RT-PCR) and hair growth in a shaving animal model of C57BL/6 mice topically applying with an amount of 0.1 mL once a day for 3 weeks. The mice were divided into 4 groups including normal group (saline, N), negative control group (dimethyl sulfoxide, NC), positive control group (5 mg/mL minoxidil, PC), and experimental group (Origanum vulgare extracts, OV). Treatment of OV didn't show cytotoxicity in HHDPC up to 10 ${\mu}g/mL$ and exhibited antioxidant activity with $IC_{50}$ of 31.0 ${\mu}g/mL$. IGF-1 expression in the skin was significantly (p<0.05) increased in the PC and OV compared to the N or NC. PC and OV also showed a prominently promoted hair regrowth compared to the N or NC in hair growth observation. The hair regrowth of OV was significantly higher than that of PC (p<0.05). Therefore, these results indicate that O. vulgare extracts effectively stimulated hair growth in an animal model.

Isolation and Identification of Short Term Drought-Induced Genes in Zea mays L. Leaves

  • Rahman, Md. Atikur;Lee, Sang-Hoon;Choi, Gi Jun;Ji, Hee Jung;Kim, Won Ho;Lee, Ki-Won
    • 한국초지조사료학회지
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    • 제37권3호
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    • pp.237-241
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    • 2017
  • Drought is one of the detrimental factors that impair plant growth and productivity. In this study, we applied annealing control primer (ACP)-based reverse transcriptase PCR (polymerase chain reaction) technique to identify differentially expressed genes (DEGs) in maize leaves in response to drought stress. Two-week-old maize seedlings were exposed to drought (DT) by suspending water supply. DEGs were screened after 3 days of DT-treated samples using the ACP-based technique. Several DEGs encoding 16.9 protein, antimicrobial protein, hypothetical protein NCLIV_068840, thioredoxin M-type were identified in maize leaves under drought stress. These genes have putative functions in plant defense response, growth and development. These identified genes would be useful for predictive markers of plant defense, and growth responses under drought stress in plants.

A Minimally Invasive Rabbit Model of Progressive and Reproducible Disc Degeneration Confirmed by Radiology, Gene Expression, and Histology

  • Kwon, Young-Joon
    • Journal of Korean Neurosurgical Society
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    • 제53권6호
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    • pp.323-330
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    • 2013
  • Objective : To develop a simple, reproducible model of disc degeneration in rabbits through percutaneous annular puncture and to confirm the degree of degeneration over time. Methods : Fifteen New Zealand white rabbits (4 to 5 months old and weighing approximately 3 to 3.5 kg each) underwent annular puncture of the L2-L3, L3-L4, and L4-L5 discs. Rabbits were sacrificed at 4, 8, or 20 weeks after puncture. For a longitudinal study to assess changes in disc height over time, serial X-rays were performed at 0, 2, 4, 8, and 20 weeks for rabbits in the 20-week group. Upon sacrifice, the whole spinal column and discs were extracted and analyzed with magnetic resonance imaging (MRI), real time reverse transcriptase-polymerase chain reaction, and histological staining. Results : The X-rays showed a slow, progressive decrease in disc height over time. Significant disc space narrowing compared to preoperative disc height was observed during the time period (p<0.001). The MRI grade, aggrecan, and matrix metalloprotease-13 mRNA expression and hematoxylin and eosin/safranin O/anti-collagen II staining were consistently indicative of degeneration, supporting the results of the X-ray data. Conclusion : Percutaneous annular puncture resulted in slow, reproducible disc degeneration that was confirmed by radiology, biochemistry, and histology. This in vivo model can be used to study and evaluate the safety and efficacy of biologic treatments for degenerative disc disease.

소아 크룹 환자들의 원인 바이러스에 따른 임상 양상과 입원 경과 비교 (Clinical presentation of croup in children according to causative viruses)

  • 김가은;신석원;최희정;최보금
    • Allergy, Asthma & Respiratory Disease
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    • 제6권6호
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    • pp.290-294
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    • 2018
  • Purpose: We evaluated the clinical features of croup in children according to viral etiology. Methods: This study enrolled pediatric patients with croup, who showed positive results on respiratory virus reverse transcriptase polymerase chain reaction performed between January 2012 and December 2017. We retrospectively reviewed the medical records. Results: A total of 179 patients (119 boys and 60 girls) were enrolled with the mean age of $18.9{\pm}14.7$ months. The viruses commonly identified were parainfluenza, respiratory syncytial virus, rhinovirus, and influenza. Among these 4 viruses, patients with rhinovirus infection showed significantly shorter fever and admission durations. Patients with parainfluenza infection showed significantly lower incidences of epinephrine nebulization and patients with influenza infections showed significantly higher incidences of steroid treatment. Conclusion: Clinical manifestations of croup differ according to causative viruses. Further studies should be conducted to evaluate the severity and prognosis of croup according to viral etiology.

Sandalwood Essential Oil의 iNOS 발현과 염증성사이토카인의 생성에 미치는 영향 (Effects of Sandalwood Essential Oil on the iNOS Expression and Proinflammatory Cytokine Production)

  • 박정숙;정숙희
    • 약학회지
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    • 제57권1호
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    • pp.70-75
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    • 2013
  • The present study was designed to determine the effect of the Sandalwood Essential Oil (Santalum album) on pro-inflammatory factors such as NO, iNOS expression and IL-$1{\beta}$, IL-6, TNF-${\alpha}$ in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of Sandalwood Essential Oil, amount of NO was measured using the NO detection kit and the iNOS expression was measured by western blot analysis and reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, Sandalwood Essential Oil reduced NO, iNOS expression and IL-$1{\beta}$, IL-6, TNF-${\alpha}$ production without cytotoxicity. Our results suggest that the Sandalwood Essential Oil may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful as an anti-inflammatory oil.

Xylitol stimulates saliva secretion via muscarinic receptor signaling pathway

  • Park, Eunjoo;Na, Hee Sam;Jeong, Sunghee;Chung, Jin
    • International Journal of Oral Biology
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    • 제44권2호
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    • pp.62-70
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    • 2019
  • Xylitol is well-known to have an anti-caries effect by inhibiting the replication of cariogenic bacteria. In addition, xylitol enhances saliva secretion. However, the precise molecular mechanism of xylitol on saliva secretion is yet to be elucidated. Thus, in this study, we aimed to investigate the stimulatory effect of xylitol on saliva secretion and to further evaluate the involvement of xylitol in muscarinic type 3 receptor (M3R) signaling. For determining these effects, we measured the saliva flow rate following xylitol treatment in healthy individuals and patients with dry mouth. We further tested the effects of xylitol on M3R signaling in human salivary gland (HSG) cells using real-time quantitative reverse-transcriptase polymerase chain reaction, immunoblotting, and immunostaining. Xylitol candy significantly increased the salivary flow rate and intracellular calcium release in HSG cells via the M3R signaling pathway. In addition, the expressions of M3R and aquaporin 5 were induced by xylitol treatment. Lastly, we investigated the distribution of M3R and aquaporin 5 in HSG cells. Xylitol was found to activate M3R, thereby inducing increases in $Ca^{2+}$ concentration. Stimulation of the muscarinic receptor induced by xylitol activated the internalization of M3R and subsequent trafficking of aquaporin 5. Taken together, these findings suggest a molecular mechanism for secretory effects of xylitol on salivary epithelial cells.

Circulating HOTAIR LncRNA Is Potentially Up-regulated in Coronary Artery Disease

  • Avazpour, Niloofar;Hajjari, Mohammadreza;Yazdankhah, Saeed;Sahni, Azita;Foroughmand, Ali Mohammad
    • Genomics & Informatics
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    • 제16권4호
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    • pp.25.1-25.5
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    • 2018
  • Coronary artery disease (CAD) is one of the leading causes of death and disability all around the world. Recent studies have revealed that aberrantly regulated long non-coding RNA (lncRNA) as one of the main classes of cellular transcript plays a key regulatory role in transcriptional and epigenetic pathways. Recent reports have demonstrated that circulating lncRNAs in the blood can be potential biomarkers for CAD. HOTAIR is one of the most cited lncRNAs with a critical role in the initiation and progression of the gene expression regulation. Recent research on the role of the HOTAIR in cardiovascular disease lays the basis for the development of new studies considering this lncRNA as a potential biomarker and therapeutic target in CAD. In this study, we aimed to compare the expression of HOTAIR lncRNA in the blood samples of patients with CAD and control samples. The expression level was examined by semi-quantitative reverse transcriptase polymerase chain reaction technique. Our data shows that expression of HOTAIR is up-regulated in blood samples of patients with CAD.