• Title/Summary/Keyword: specific RNA

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Secretion of MCP-1, IL-8 and IL-6 Induced by House Dust Mite, Dermatophagoides pteronissinus in Human Eosinophilic EoL-1 Cells

  • Lee, Ji-Sook;Kim, In-Sik;Yun, Chi-Young
    • Animal cells and systems
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    • v.13 no.4
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    • pp.391-397
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    • 2009
  • The house dust mite (Dermatophagoides pteronissinus) is an important factor in triggering allergic diseases. The function of eosinophils, particularly in the production of cytokine or chemokine, is critical in understanding the pathogenesis of inflammatory diseases. In this study, we examined whether D. pteronissinus extract (DpE) induces the expression of monocyte chemotactic protein 1 (MCP-1)/CCL2, IL-8/CXCL8, and IL-6 that mediate in the infiltration and activation of immune cells and in its signaling mechanism in the human eosinophilic cell line, EoL-1. DpE increased the mRNA and protein expression of MCP-1, IL-8, and IL-6 in a time- and dose-dependent course in EoL-1 cells. In our experiments using signal-specific inhibitors, we found that the increased expression of MCP-1, IL-8, and IL-6 due to DpE is associated with Src family tyrosine kinase and protein kinase C $\delta$ (PKC $\delta$). In addition, the activation of extracellular signal-regulated kinase (ERK) is required for MCP-1 and IL-8 expression while p38 mitogen-activated protein kinase (MAPK) is involved in IL-6 expression. DpE induced the phosphorylation of ERK and p38 MAPK. PP2, an inhibitor of Src family tyrosine kinase, and rottlerin, an inhibitor of PKC $\delta$, blocked the activation of ERK and p38 MAPK. DpE induces the activation of ERK and p38 MAPK via Src family tyrosine kinase and PKC $\delta$ for MCP-1, IL-8, or IL-6 production. Increased cytokine release due to the house dust mite and the characterization of its signal transduction may be valuable in understanding the eosinophil-related pathogenic mechanism of inflammatory diseases.

Molecular Authentication of Magnoliae Flos Using Robust SNP Marker Base on trnL-F and ndhF Region

  • Kim, Min-Kyeoung;Noh, Jong-Hun;Yan, Deok-Chun;Lee, Sanghun;Lee, Hee-Nyeong;Jin, Chi-Gyu
    • Korean Journal of Plant Resources
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    • v.28 no.3
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    • pp.341-349
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    • 2015
  • Magnoliae Flos (Sini in Korean) is one of the most important oriental medicinal plants. In the Korean Herbal Pharmacopeia, the bud of the all species in Manolia denudate and Manolia genus were regarded as the botanical sources for ‘Sini’. Most the dried bud of Manolia denudata, Manolia biondii and Manolia sprengeri were used as ‘Xin-yi’ in China. Therefore, the purpose of this study was to determine and compare the ‘Magnolia’ species, four species including Manolia denudata, M. biondii, M. liliiflora and M. Kobus were analysis of sequencing data revealed DNA polymorphisms. The based on tRNA coding leucine/phenylalanine (trnL-F) and NADH-plastoquinone oxidoreductase subunit 5 (ndhF) sequences in chloroplast DNA. For the identification of ‘Magnolia’ species, polymerase chain reaction (PCR) analysis of chloroplast DNA regions such as ndhF have proven an appropriate method. A single nucleotide polymorphism (SNP) has been identified between genuine “Sini” and their fraudulent and misuse. Specific PCR primers were designed from this polymorphic site within the sequence data, and were used to detect true plants via multiplex PCR.

Anti-Cancer Activity of the Flower Bud of Sophora japonica L. through Upregulating Activating Transcription Factor 3 in Human Colorectal Cancer Cells

  • Lee, Jin Wook;Park, Gwang Hun;Eo, Hyun Ji;Song, Hun Min;Kim, Mi Kyoung;Kwon, Min Ji;Koo, Jin Suk;Lee, Jeong Rak;Lee, Man Hyo;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.28 no.3
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    • pp.297-304
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    • 2015
  • The flower buds of Sophora japonica L (SF), as a well-known traditional Chinese medicinal herb, have been used to treat bleeding-related disorders such as hematochezia, hemorrhoidal bleeding, dysfunctional uterine bleeding, and diarrhea. However, no specific anti-cancer effect and its molecular mechanism of SF have been described. Thus, we performed in vitro study to investigate if treatment of SF affects activating transcription factor 3 (ATF3) expression and ATF3-mediated apoptosis in human colorectal cancer cells. The effects of SF on cell viability and apoptosis were measured by MTT assay and Western blot analysis against cleaved poly (ADP-ribose) polymerase (PARP). ATF3 activation induced by SF was evaluated using Western blot analysis, RT-PCR and ATF3 promoter assay. SF treatment caused decrease of cell viability and increase of apoptosis in a dose-dependent manner in HCT116 and SW480 cells. Exposure of SF activated the levels of ATF3 protein and mRNA via transcriptional regulation in HCT116 and SW480 cells. Inhibition of extracellular signal-regulated kinases (ERK) 1/2 by PD98059 and p38 by SB203580 attenuated SF-induced ATF3 expression and transcriptional activation. Ectopic ATF3 overexpression accelerated SF-induced cleavage of PARP. These findings suggest that SF-mediated apoptosis may be the result of ATF3 expression through ERK1/2 and p38-mediated transcriptional activation.

PREVALENCE OF BLACK-PIGMENTED BACTERIA IN INFECTED ROOT CANALS IN KOREA (감염 근관의 흑색세균의 동정)

  • Chung, Ki-Soo;Lim, Sung-Sam;Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.24 no.3
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    • pp.447-452
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    • 1999
  • The role of bacteria in root canals and periapical infections is well known and established. In these bacteria, black-pigmented bacteria(BPH) play important role in endodontic infection. BPB are Gram negative anaerobic rods which are closely related 50 clinical symptoms such as pain, percussion, tenderness, foul odor, etc. In America and Europe, many studies on BPB have been done and are continued. But, relatively few studies have been done in Korea, especially its prevalence in Korean population is not yet studied. The purpose of this study is to establish prevalence of BPB in infected root canals and periapical abscesses in Korean people. Microbial samples were collected from the root canals of 34 intact tooth with periapical rarefactions of endodontic origin and 3 periapical abscesses. All samples were incubated in an anaerobic chamber(Coy, Model No. 77. Ann Arbor, Michigan, USA.). Identification of In microorganism was based on its growth in the anaerobic chamber, colonial pigmentation, colonial morphology, Gram stain, and Rapid ID32A(BioMericux SA/69280 Marcy-l'Etoile/France) results. In addition, the polyme ase chain reaction using specific primers for 16S rRNA genes were used differentiate Prevotella nigrescens for Prevotella intermedia. The results were as follows : 1. In this study, thirteen (35%) of thirty seven samples were positive for the growth of BPB. In thirteen samples, sixteen strains of BPR were found. 2. The most frequently identified BPB in root canals and abscesses in Korean were P. nigrescens 5/37(14%) and P. intermedia 5/37(14%). Porphyromonas gingivalis 3/37(8%), Porphyromonas endodontalis 2/37(5%) and Prevotella loecheii 1/37(3%) were also found. 3. In this study, no significant differences were found between the prevalence of BPB in Korean and that of American and European.

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Phylogenetic Analysis and Rapid Detection of Genus Phellinus using the Nucleotide Sequences of 18S Ribosomal RNA

  • Nam, Byung-Hyouk;Lee, Jae-Yun;Kim, Gi-Young;Jung, Heon-Ho;Park, Hyung-Sik;Kim, Cheng-Yun;Jo, Wol-Soon;Jeong, Soo-Jin;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.31 no.3
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    • pp.133-138
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    • 2003
  • Analysis of phylogenetic relationship was performed among Phellinus species based on 18S ribosomal subunit sequence data. Twenty-five strains of 19 Phellinus species including P. linteus were examined in this study. Regions of 18S ribosomal subunit were very conserved, but some variable regions between Phellinus species were observed. The species-specific detection primers, modified by 2 or 3 nucleotides in sense primer were designed based on 18S ribosomal DNA(rDNA) sequence data. The 210 by PCR bands were detected with annealing temperature $48^{\circ}C$. The 18S 2F-18S 4R detection primer set distinguished P. linteus from various Phellinus species but some species like P. baumii, P. weirianius, P. rhabarberinus and P. pomaceus also had weak reactivity on this primer set. The 18S 3F-18S 4R primer set distinguished only P. linteus from various Phellinus species, although sensitivity with this primer set was lower than that of 18S 2F-18 4R primer set. These primer sets would be useful for the detection of only P. linteus among unknown Phellinus species rapidly.

Evaluation of Japanese encephalitis virus vaccine strains currently used in pigs by molecular characterization

  • Lee, Jeong-Ah;Yang, Dong-Kun;Kim, Ha-Hyun;Kim, Sun-Young;Nah, Jin-Ju;Cho, Soo-Dong;Song, Jae-Young
    • Korean Journal of Veterinary Service
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    • v.35 no.3
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    • pp.169-174
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    • 2012
  • Japanese encephalitis virus (JEV) is one of the main causes of viral encephalitis in human and animals. For over 30 years, a live attenuated JEV vaccine strain has been used in the veterinary field, and it is required to conduct quality evaluation studies on the commercial vaccines. For the quality control of live attenuated JEV vaccine, we investigated the nucleotide sequence similarity of prME gene derived from five JEV vaccines commercially available in pigs in Korea. The Vero cells infected with JEV vaccines showed specific cytopathic effect, which was characterized by rounding and detached cells. In the phylogenetic analysis, all of the vaccine strains showed a close relationship with the original vaccine seed strain (Anyang 300) and clustered into the genotype 3. In comparison of the nucleotide and deduced amino acid sequences of prME genes with the original strain, all JEV vaccine strains showed high amino acid similarity ranging from 98.9% to 99.5%, but had several point mutations, probably due to high mutation rates of viral RNA polymerase by several virus passages. Even though the current JEV vaccine strains have been maintained and produced for a long period of time, the genetic characterization of them have been rarely changed. However, since the mid 1990's, molecular epidemiology of JEV has been changed sharply from genotype 3 to genotype 1 in Korea, further studies on new vaccine strains to genotype 1 is required for more effective prevention in the field.

Isolation of Brucella spp from sere-positive native bulls and calves below twelve months old (브루셀라병 혈청검사 양성 수소와 12개월령 이하 소에서의 균 분리 및 동정)

  • Ryu, Jae-Yun;Bun, Jung-Hyun;Lee, Hee-Young;Lee, Yong-Chang;Lee, Jong-Jin;Song, Young-Gak;Nam, Hyang-Mi
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.375-384
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    • 2007
  • This study was conducted to investigate the characteristics of brucellosis in Korean native cattle in a farm where bovine brucellosis was confirmed 3 times from September 2006 to March 2007. Of 74 bulls serum samples examined, 21 (28.4%) were positive by Rose-Bengal test (RBT) and Standard tube agglutination test (STAT). In the isolation test from seropositive bulls, B abortus was isolated and identified from 2 specimens (testis, intestinal lymph node) among 6 kinds of specimens including blood, urine, feces and soil. Isolation rate of intestinal lymph node and testis was 25% (3/12 cases) and 16.7% (2/12), respectively. B abortus was also isolated from calves below 12 months old, i.e., 1 isolate (25.0%) was confirmed from testis, 4 (40.0%) from supra-mammary lymph nodes and 1 (25.0%) from intestinal lymph node. All isolates had Brucella specific 16s r-RNA with 905-bp band detected by PCR assay. For the more effective control of bovine brucellosis in korea, this paper would like to suggest that all of bulls and calves should be included in the screening tests.

Antiadipogenic Effects of Salvia plebeia R. Br. Extracts by Extraction Conditions in 3T3-L1 Preadipocytes (추출조건에 따른 배암차즈기 추출물의 지방세포 분화 및 지방축적 억제 효과)

  • Choi, Soo Im;Kwak, Hoyoung;Kim, Jae Yoon;Choi, Jong Gil;Lee, Je Hyuk
    • Korean Journal of Medicinal Crop Science
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    • v.23 no.3
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    • pp.245-252
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    • 2015
  • This study was carried out to investigate the effects of Salvia plebeia R. Br. ethanolic extract with different aspects (stem/leaf and whole plant) on differentiation and lipid accumulation in 3T3-L1 preadipocytes. The morphological changes and the degrees of lipid accumulation in 3T3-L1 cells were measured by Oil Red O staining and intra-cellular triglyceride (TG) assay. The mRNA expressions of special peroxisome proliferation activated receptor- genes (PPAR), CCAAT/ enhancer-binding protein (C/$EBP{\alpha}$), fatty acid synthase (FAS) and lipoprotein lipase (LPL) were detected by reverse transcriptase polymerase chain reaction (RT-PCR). The 50% ethanolic extracts ($100{\mu}g/mL$) of stem and leaf (SALE) and 30% ethanolic extracts (100 g/mL) of whole plant (SAE) from Salvia plebeia R. Br. were significantly attenuated lipid accumulation during adipogenesis in 3T3-L1 cells. Ethyl acetate-soluble fractions ($50{\mu}g/mL$) significantly inhibited lipid droplet accumulation in 3T3-L1 cells. In addition, SALE induced down-regulation of specific adipogenic transcriptional factors (C/$EBP{\alpha}$ and $PPAR{\gamma}$) and target genes (FAS and LPL) during adipogenesis. Salvia plebeia R. Br. may be used as a safe and efficient natural substance to manage obesity.

Predictive value of C-reactive protein in response to macrolides in children with macrolide-resistant Mycoplasma pneumoniae pneumonia

  • Seo, Young Ho;Kim, Jang Su;Seo, Sung Chul;Seo, Won Hee;Yoo, Young;Song, Dae Jin;Choung, Ji Tae
    • Clinical and Experimental Pediatrics
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    • v.57 no.4
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    • pp.186-192
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    • 2014
  • Purpose: The prevalence of macrolide-resistant Mycoplasma pneumoniae (MRMP) has increased worldwide. The aim of this study was to estimate the proportion of MRMP in a tertiary hospital in Korea, and to find potential laboratory markers that could be used to predict the efficacy of macrolides in children with MRMP pneumonia. Methods: A total of 95 patients with M. pneumoniae pneumonia were enrolled in this study. Detection of MRMP was based on the results of specific point mutations in domain V of the 23S rRNA gene. The medical records of these patients were reviewed retrospectively and the clinical course and laboratory data were compared. Results: The proportion of patients with MRMP was 51.6% and all MRMP isolates had the A2063G point mutation. The MRMP group had longer hospital stay and febrile period after initiation of macrolides. The levels of serum C-reactive protein (CRP) and interleukin-18 in nasopharyngeal aspirate were significantly higher in patients who did not respond to macrolide treatment. CRP was the only significant factor in predicting the efficacy of macrolides in patients with MRMP pneumonia. The area under the curve for CRP was 0.69 in receiver operating characteristic curve analysis, indicating reasonable discriminative power, and the optimal cutoff value was 40.7 mg/L. Conclusion: The proportion of patients with MRMP was high, suggesting that the prevalence of MRMP is rising rapidly in Korea. Serum CRP could be a useful marker for predicting the efficacy of macrolides and helping clinicians make better clinical decisions in children with MRMP pneumonia.

Bacterial Community and Biological Nitrate Removal: Comparisons of Autotrophic and Heterotrophic Reactors for Denitrification with Raw Sewage

  • Lee, Han-Woong;Park, Yong-Keun;Choi, Eui-So;Lee, Jin-Woo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1826-1835
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    • 2008
  • An autotrophic denitrification reactor (ADR-l) and a heterotrophic denitrification reactor (HDR-2) were operated to remove nitrate and nitrite in an anoxic environment in raw sewage. The $NO_3$-N removal rate of ADR-l was shown to range from 52.8% to 78.7%, which was higher than the $NO_3$-N removal rate of HDR-2. Specific denitrification rates (SDNR) of ADR-l and HDR-2 were 3.0 to 4.0 and 1.1 to $1.2\;mgNO_3$-N/gVSS/h, respectively. From results of restriction fragment length polymorphism (RFLP) of the 16S rRNA gene, Aquaspirillum metamorphum, Alcaligenes defragrans, and Azoarcus sp. were $\beta$-Proteobacteria that are affiliated with denitritying bacteria in the ADR-l. Specifically, Thiobacillus denitrificans was detected as an autotrophic denitrification bacteria. In HDR-2, the $\beta$-Proteobacteria such as Denitritying-Fe-oxidizing bacteria, Alcaligenes defragrans, Acidovorax sp., Azoarcus denitrificans, and Aquaspirillum metamorphum were the main bacteria related to denitrifying bacteria. The $\beta$-and $\alpha$-Proteobacteria were the important bacterial groups in ADR-l, whereas the $\beta$-Proteobacteria were the main bacterial group in HDR-2 based on results of fluorescent in situ hybridization (FISH). The number of Thiobacillus denitrificans increased in ADR-l during the operation period but not in HRD-2. Overall, the data presented here demonstrate that many heterotrophic denitritying bacteria coexisted with autotrophic denitrifying bacteria such as Thiobacillus denitrificans for nitrate removal in ADR-l. On the other hand, only heterotrophic denitritying bacteria were identified as dominant bacterial groups in HDR-2. Our research may provide a foundation for the complete nitrate removal in raw sewage of low-COD concentration under anoxic condition without any external organic carbon or the requirement of post-treatment.