• 제목/요약/키워드: Direct PCR

검색결과 410건 처리시간 0.037초

이프리플라본을 함유한 생분해성 PLGA 미립구의 제조 및 조직공학적 골재생을 위한 영향평가 (Preparation and Release Behavior of Ipriflavone-Loaded PLGA Microsphere for Tissue Engineered Bone)

  • 소정원;장지욱;김순희;김근아;최진희;이종문;손영숙;민병현;강길선
    • 폴리머
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    • 제33권1호
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    • pp.26-32
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    • 2009
  • 골 결손 치료를 위해 생분해성 고분자인 PLGA에 골다공증 치료제인 이프리플라본(IP)을 함유한 미립구를 O/W 유화 용매 증발법으로 제조하였으며, 생체외 방출실험에서 IP 방출량은 HPLC로 분석하였다. SEM을 이용하여 미립구에 부착된 세포의 거동을 확인하였으며, IP가 세포에 미치는 독성평가는 CCK-8 분석방법을 이용하여 측정하였다. 골 형성 지표인 ALP 활성도를 측정하였으며, 배양된 BMSCS의 골세포로의 표현형을 확인하기 위하여 RT-PCR을 수행하였다. 방출결과에서 IP 방출은 거의 40일 이상으로 지속적이었으며, IP를 함유한 미립구에서의 세포의 부착, 성장 등이 잘 이루어짐을 확인하였고, ALP 활성도 및 RT-PCR 분석결과에서도 단독의 PLGA 미립구보다 IP를 함유한 미립구에서의 값이 더 높은 것을 확인할 수 있었다. 본 실험결과를 바탕으로 향후 서방성 제제화에 있어서 IP/PLGA 미립구를 응용하게 되면 국소지향성 골분화 주사용 지지체로서 중요한 역할을 할 것으로 보인다.

수계환경에서 분변성 오염의 지표로 사용되는 미생물들 (Indicator Microorganisms Used as Fecal Contamination in Aquatic Environments)

  • 이건형
    • 환경생물
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    • 제20권3호
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    • pp.189-196
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    • 2002
  • The direct detection of intestinal pathogens and viruses often requires costly, tedious, and time-consuming procedures. These requirements developed a test to show that the water was contaminated with sewage-borne pathogens by assessing the hygienic quality of water based on indicator microorganisms whose presence indicates that pathogenic microorganisms may also be present. Various groups of microorganisms have been suggested and used as indicator microorganisms. Proposed and commonly used microbial indicators are total coliforms, fecal coliforms, fecal streptococci, Clostridium perfringens, heterotrophic plate count, bacteriophage, and so on. Unfortunately, most, if not all, of these indicators are not ideal because of the sensitivity and resistance to environment stresses and disinfection. However, the development of gene probes and PCR technology may give hope for the discovery of rapid and simple methods toy detecting small number of fecal pathogens in various environments.

Sequence Analysis of Cochlodinium polykrikoides Isolated from Korean Coastal Waters Using Sequences of Internal Transcribed Spacers and 5.8S rDNA

  • Kim, Hak-Gyoon;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • 제35권3호
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    • pp.158-160
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    • 2000
  • The relativity of four isolates of C. polykrikoides was determined by comparative sequence analysis based on direct sequencing of PCR amplified ribosomal DNA (the internal transcribed spacer region and the 5.8S rDNA). Sequence comparisons indicated that four isolates had the same nucleotide sites in the ITS regions, as well as a total of 585 nucleotide length and 100% homology. The molecular data revealed that C. polykrikoides in Korean coastal waters show no genetical difference.

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The effect of Astragali Radix Ethanol extract on Murine CD4 T cells′ Cytokine Profiles in vitro

  • Hee Kang;Bae Hyun Su;Ahn Kyoo Seok
    • 동의생리병리학회지
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    • 제17권5호
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    • pp.1330-1334
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    • 2003
  • Astragali Radix(AR), one of the strong tonic herbs, is known to improve immunological responses in mice and human. In this study, AR's ai-reinforcing effect was examined in the context of CD4/sup +/ T cells' TCR/CD3 induced activation responses. In order to evaluate the direct effect of AR on helper T cells, CD4/sup +/ T cells are isolated using magnetic bead and their proliferation and CD69 expression in AR treated medium were assessed with anti-CD3/anti-CD28 activation for 48h. CD4 T cells' proliferation was slightly increased but there was little effect on CD69 expression. RT PCR and ELISA equally demonstrated that IL-2 and IL-4 production was increased but IFN-ν was down-regulated. This shows AR ethanol extract favors Th2 cytokine profile under neutral conditions.

Human rhinoviruses and asthma in children

  • Kim, Woo Kyung
    • Clinical and Experimental Pediatrics
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    • 제53권2호
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    • pp.129-135
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    • 2010
  • Human rhinoviruses (HRVs) is a nonenveloped, single stranded RNA virus belonging to the family Picornavirudae. Transmission by direct contact such as hand-to-hand, hand-to-nose, and hand-to-eye has been readily demonstrated in experimental settings. HRV are the most frequent causes of common cold infection, however, they are also known to replicate in the lower respiratory tract and associated with more severe respiratory illnesses such as asthma. New technique such as reverse transcriptase polymerase chain reaction and molecular typing in HRV has been developed and our understanding of the importance of these respiratory viruses. HRVs consisted of 101 serotypes that are classified into groups A and B according to sequence variations. And there is a newly identified set of HRVs, called Group C, and it is currently under investigation. In recent study using PCR techniques, HRVs accounted for approximate 50-80% of common colds and 85 % of childhood asthma exacerbations and in more than half of adult exacerbations. However, the mechanisms of HRV- induced asthma exacerbations are poorly understood. This review discusses the association between HRVs and childhood asthma.

Sequence-specific interaction between ABD-B homeodomain and castor gene in Drosophila

  • Kim, Keon-Hee;Yoo, Siuk
    • BMB Reports
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    • 제47권2호
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    • pp.92-97
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    • 2014
  • We have examined the effect of bithorax complex genes on the expression of castor gene. During the embryonic stages 12-15, both Ultrabithorax and abdominal-A regulated the castor gene expression negatively, whereas Abdominal-B showed a positive correlation with the castor gene expression according to real-time PCR. To investigate whether ABD-B protein directly interacts with the castor gene, electrophoretic mobility shift assays were performed using the recombinant ABD-B homeodomain and oligonucleotides, which are located within the region 10 kb upstream of the castor gene. The results show that ABD-B protein directly binds to the castor gene specifically. ABD-B binds more strongly to oligonucleotides containing two 5'-TTAT-3' canonical core motifs than the probe containing the 5'-TTAC-3' motif. In addition, the sequences flanking the core motif are also involved in the protein-DNA interaction. The results demonstrate the importance of HD for direct binding to target sequences to regulate the expression level of the target genes.

Iridovirus의 해산 양식어류에 대한 병원성과 사육수에서의 검출 (Pathogenicity of Iridovirus against Marine Fish and Its Detection in Culturing Seawater)

  • 정준범;정현도
    • 한국수산과학회지
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    • 제41권1호
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    • pp.20-25
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    • 2008
  • The susceptibility of five different marine fish to iridovirus IVS-1 infection was analyzed and found a higher the cumulative mortality in the order of rock bream (Oplegnathus fasciatus), red sea bream (Pagrus major), sea perch (Lateolabrax sp.), rockfish (Sebastes schlegeli) and black porgy (Acanthopagrus schlegeli). However, the concentrations of virus in the infected spleens of these species did not differ significantly. To determine the release of iridovirus from infected fish into culturing seawater, rock bream were challenged with iridovirus IVS-1 and the concentration of virus in the water was analyzed using PCR. Over the 10 days of the analysis, the linear relationship between the number of dead fish and viral DNA concentration found in culturing seawater should be considered direct evidence of horizontal iridovirus transmission.

Vascular Endothelial Growth Factor Upregulates Follistatin in Human Umbilical Vein Endothelial Cells

  • Oh, In-Suk;Kim, Hwan-Gyu
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권3호
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    • pp.201-206
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    • 2004
  • Vascular endothelial growth factor (VEGF), plays a key role in angiogenesis. Many endogenous factors can affect angiogenesis in endothelial cells. VEGF is known to be a strong migration, sprouting, survival, and proliferation factor for endothelial cells during angiogenesis in endothelial cells. Searching for novel genes involved in VEGF signaling during angiogenesis, we carried out differential display polymerase chain reaction on RNA from VEGF-stimulated human umbilical vein endothelial cells (HUVECs). In this study, follistatin (FS) differentially expressed in VEGF-treated HUVECs, compared with controls. Addition of VEGF (10ng/L) produced an approximately 11.8-fold increase of FS mRNA. F5 or VEGF produced approximately 1.8- or 2.9-fold increases, respectively, in matrix metalloproteinase-2 (MMP-2) secretion for 12h, compared to the addition of a control buffer. We suggest that VEGF may affect the angiogenic effect of HUVECs, through a combination of the direct effects of VEGF itself, and the indirect effects mediated via induction of FS in vitro.

Characterization of the Nucleotide Sequence of a Polyubiquitin Gene (PUBC1) from Arabian Camel, Camelus dromedarius

  • Al-Khedhairy, Abdulaziz Ali A.
    • BMB Reports
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    • 제37권2호
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    • pp.144-147
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    • 2004
  • Molecular amplification and sequencing of genomic DNA that encodes camel polyubiquitin (PUBC1) was performed by a polymerase chain reaction (PCR) using various sets of primers. The amplification generated a number of DNA fragments, which were sequenced and compared with the polyubiquitin coding sequences of various species. One DNA fragment that conformed to 325 bp was found to be 95 and 88% homologous to the sequences of human polyubiquitin B and C, respectively. The DNA translated into 108 amino acids that corresponded to two fused units of ubiquitin with no intervening sequence, which indicates that it is a polyubiquitin and contains at least two units of ubiquitin. Although, variations were found in the nucleotide sequence when compared to those of other species, the amino acid sequence was 100% homologous to the polyubiquitin sequences of humans, mice, and rats. This is the first report of the polyubiquitin DNA coding sequence and its corresponding amino acid sequence from camels, amplified using direct genomic DNA preparations.

CHARACTERISTICS OF BIOHYDROGEN PRODUCTION AND MICROBIAL COMMUNITY AS A FUNCTION OF SUBSTRATE CONCENTRATION

  • Youn, Jong-Ho;Shin, Hang-Sik
    • Environmental Engineering Research
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    • 제10권1호
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    • pp.7-14
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    • 2005
  • The feasibility of hydrogen production with a raw seed sludge through direct acclimation of feedstock was investigated at acidogenic stage, and methane was harvested at followed methanogenic stage in an anaerobic two-stage process. Hydrogen content was higher than 57% at all tested organic loading rates (OLRs) and the yield of hydrogen ranged from 1.5 to 2.4 mol H2/mol hexose consumed and peaked at 6 gVSl-1day-1. Normal butyrate and acetate were main volatile fatty acids (VFAs), whereas the concentration of propionate was insignificant. The hydrogen-producing bacteria, Clostridium thermosaccharolyticum, was detected with strong intensity at all tested organic loading rates (OLRs) by denaturing gradient gel electrophoresis (DGGE) of the polymerase chain reaction (PCR) analysis. From COD balance in the process, the fraction of the feed-COD converted to the hydrogen-COD at acidogenic stage ranged from 7.9% to 9.3% and peaked at 6 gVSl-1day-1, whereas the fraction of feed-COD converted to the methane-COD at methanogenic stage ranged from 66.2% to 72.3% and peaked at 3 gVSl-1day-1.