• Title/Summary/Keyword: Target DNA

Search Result 786, Processing Time 0.035 seconds

Effect of Ginsenoside on Basal and Nitro-L-Arginine Suppressed Nitric Oxide Production in Rat Kidney

  • Kim, Hye-Young;Han, Sang-Won
    • Biomolecules & Therapeutics
    • /
    • v.2 no.2
    • /
    • pp.131-135
    • /
    • 1994
  • The effect of ginsenoside (GS) from Panax ginseng on basal and nitro-L-arginine suppressed nitric oxide (NO) production was studied in rat kidney. NO production was determined by conversion to [$^{14C}$]=L-citrulline from [$^{14C}$]-L-arginine both in whole kidney and three renal segments; glomerulus, cortex excluding glomerulus (cortex-) and medulla. Nitro-L-arginine (total dose of 30 mg/kg/3 days, i.p.) significantly reduced NO production in whole kidney, which was prevented by GS pretreatment (30 mg/kg/3 days, i.p.). Relative high dose of GS (120 mg/kg/4 days, i.p..) selectively increased NO production in glomerulus and cortex-. Protein content, on wet weight basis, in cortex- and glomerular DNA content were significantly reduced by GS. Our results confirm the existence of constitutive nitric oxide synthase in kidney and it seems that target nephron segment for volume expansion due to GS'NO-mediated vasodilation and for NO production stimulated by GS is cortex including glomerulus.lus.

  • PDF

ent-Kaurane Diterpenoids from Croton tonkinensis Inhibit LPS-induced Transcription Factor NF-${\kappa}{B}$ Activation and NO Production

  • Giang, Phan-Minh;Jin, Hui-Zi;Lee, Jung-Joon
    • Proceedings of the PSK Conference
    • /
    • 2003.04a
    • /
    • pp.120.1-120.1
    • /
    • 2003
  • Nuclear factor-${\kappa}{B}$ (NF-${\kappa}{B}$) belongs to a group of homodimers and heterodimers of Rel/NF-${\kappa}{B}$ proteins that bind to DNA target sites, where they directly regulate gene transcription. The activation of NF-${\kappa}{B}$ has been shown to mediate inflammation and suppress apoptosis. Activated NF-${\kappa}{B}$ has been found n various inflammatory diseases such as rheumatoid arthritis, Atherosclerosis, asthma, nflammatory bowel disease, and Helicobacter pylori-associated gastritis and associated with cancer, cachexia, diabetes, euthyroid sick syndrome, and AIDS. (omitted)

  • PDF

Design and Synthesis of N-Aryl 8,9-Dihydro-7H-isoindolo[5,6-g]quinoxaline-7,9-dione Derivatives as Potential Antitumor Agent

  • Lee, Hee-Soon;Jung, Eun-Kyung;Nam, Koong-Kwon;Jung, Jae-Kyung
    • Proceedings of the PSK Conference
    • /
    • 2003.10b
    • /
    • pp.174.1-174.1
    • /
    • 2003
  • We have previously reported the synthesis and cytotoxic activities of a series of azaanthraquinone derivatives using doxorubicin as a lead compound. Doxorubicin is known to intercalate into DNA and to inhibit topoisomerase II activity. But in the case of quinone compounds like Dox, its use is limited because of systemic toxicities, primarily cardiotoxicity and myelosuppression. In this study, we discuss the synthesis of isoindolobenzoquinoxaline derivatives. The quinone group of the azaanthraquinone derivatives were removed in the target compounds. (omitted)

  • PDF

Development of Diagnostic Technology of Xylella fastidiosa Using Loop-Mediated Isothermal Amplification and PCR Methods

  • Kim, Suyoung;Park, Yujin;Kim, Gidon
    • Research in Plant Disease
    • /
    • v.27 no.1
    • /
    • pp.38-44
    • /
    • 2021
  • Xylella fastidiosa is the most damaging pathogen in many parts of the world. To increase diagnostic capability of X. fastidiosa in the field, the loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assay were developed to mqsA gene of citrate-synthase (XF 1535) X. fastidiosa and evaluated for specificity and sensitivity. Both assays were more robust than current published tests for detection of X. fastidiosa when screened against 16 isolates representing the four major subgroups of the bacterium from a range of host species. No cross reaction with DNA from healthy hosts or other species of bacteria has been observed. The LAMP and PCR assays could detect 10-4 pmol and 100 copies of the gene, respectively. Hydroxynaphthol blue was evaluated as an endpoint detection method for LAMP. There was a significant color shift that signaled the existence of the bacterium when at least 100 copies of the target template were present.

Detection of the Causal Agent of Bacterial Wilt, Ralstonia solanacearum in the Seeds of Solanaceae by PCR (가지과 종자에서 Ralstonia solanacearum의 검출을 위한 PCR 방법)

  • Cho, Jung-Hee;Yim, Kyu-Ock;Lee, Hyok-In;Baeg, Ji-Hyun;Cha, Jae-Soon
    • Research in Plant Disease
    • /
    • v.17 no.2
    • /
    • pp.184-190
    • /
    • 2011
  • Ralstonia solanacearum, a causal agent of bacterium wilt is very difficult to control once the disease becomes endemic. Thus, Ralstonia solanacearum is a plant quarantine bacterium in many countries including Korea. In this study, we developed PCR assays, which can detect Ralstonia solanacearum from the Solanaceae seeds. Primers RS-JH-F and RS-JH-R amplified specifically a 401 bp fragment only from Ralstonia solanacearum race 1 and race 3. The nested PCR primers, RS-JH-F-ne and RS-JH-R-ne that were designed inside of 1st PCR amplicon amplified specifically a 131 bp fragment only from Ralstonia solanacearum race 1 and race 3. The primers did not amplify any non-specific DNA from the seed extracts of the Solanaceae including tomato and pepper. When detection sensitivity were compared using the Solanaceae seeds inoculated with target bacteria artificially, the nested PCR method developed in this study 100 times more sensitive than ELISA and selective medium. Therefore, we believe that the PCR assays developed in this work is very useful to detect Ralstonia solanacearum in the Solanaceae seeds.

Evaluation of MiR-34 Family and DNA Methyltransferases 1, 3A, 3B Gene Expression Levels in Hepatocellular Carcinoma Following Treatment with Dendrosomal Nanocurcumin

  • Chamani, Fatemeh;Sadeghizadeh, Majid;Masoumi, Mahbobeh;Babashah, Sadegh
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.17 no.sup3
    • /
    • pp.219-224
    • /
    • 2016
  • Hepatocellular carcinoma (HCC) is the most common primary malignancy of the liver making up more than 80 percent of cases. It is known to be the sixth most prevalent cancer and the third most frequent cause of cancer related death worldwide. Epigenetic regulation constitutes an important mechanism by which dietary components can selectively activate or inactivate target gene expression. The miR-34 family members including mir-34a, mir-34b and mir-34c are tumor suppressor micro RNAs, which are expressed in the majority of normal tissues. Several studies have indicated silencing of miR-34 expression via DNA methylation in multiple types of cancers. Bioactive nutrients like curcumin (Cur) have excellent anticarcinogenic activity and minimal toxic manifestations in biological systems. This compound has recently been determined to induce epigenetic changes. However, Cur is lipophilic and has a poor systemic bioavailability and poor absorption. Its bioavailability is increased through employing dendrosome nanoparticles. The aim of the current study was to investigate the effect of dendrosomal nanocurcumin (DNC) on expression of mir-34 family members in two HCC cell lines, HepG2 and Huh7. We performed the MTT assay to evaluate DNC and dendrosome effects on cell viability. The ability of DNC to alter expression of the mir-34 family and DNA methyltransferases (DNMT1, DNMT3A and 3B) was evaluated using semi-quantitative and quantitative PCR. We observed the entrance of DNC into HepG2 and Huh7 cells. Gene expression assays indicated that DNC treatment upregulated mir34a, mir34b and mir34c expression (P<0.05) as well as downregulated DNMT1, DNMT3A and DNMT3B expression (P<0.05) in both HepG2 and Huh7 cell lines. DNC also reduced viability of Huh7 and HepG2 cells through restoration of miR-34s expression. We showed that DNC could awaken the epigenetically silenced miR-34 family by downregulation of DNMTs. Our findings suggest that DNC has potential in epigenetic therapy of HCC.

Study on Rumen Cellulolytic Bacterial Attachment and Fermentation Dependent on Initial pH by cPCR (cPCR 기법을 이용한 초기배양 pH에 의한 반추위 섬유소 분해 박테리아의 부착 및 발효에 관한 연구)

  • Kim, M.S.;Sung, H.G.;Kim, H.J.;Lee, Sang-S.;Chang, J.S.;Ha, J.K.
    • Journal of Animal Science and Technology
    • /
    • v.47 no.4
    • /
    • pp.615-624
    • /
    • 2005
  • The cPCR technique was used to monitor rumen fermentation and attachment of Fibrobacter succinogenes to cellulose at different pH in the in vitro culture medium. The target fragments of 16S rDNA(445 bp) were amplified from genomic DNA of F. succinogenes with specific primers and internal controls(205 bp) were constructed. Cell counts were estimated from the amounts of genomic DNA, which was calculated from cPCR results. F. succinogenes in pH 6.8 and 6.2 showed apparently higher attachment than in pH 5.8 during all incubation time. There were some difference between pH 6.8 and 6.2 in the degree of attachment, but the different was not significant (P>0.05). Cellulose degradation increased in process of incubation time and the increasing rate was higher when initial pH was higher. The pH in culture medium decreased regardless of initial pH in course of incubation time. After 24 h of incubation, medium pH was dropped by 0.24, 0.58 and 0.16 units from original medium pH 6.8, 6.2 and 5.8, respectively. More gas was produced at higher initial pH in the same manner as in cellulose degradation. In summery, Initial pH of rumen culture in vitro significantly influenced cellulose digestion, gas production, pH change and bacterial attachment. Especially, low pH(5.8) resulted in much lower bacterial attachment and fiber digestion compared to higher medium pH.

Differentiation of Four Major Gram-negative Foodborne Pathogenic Bacterial Genera by Using ERIC-PCR Genomic Fingerprinting (ERIC-PCR genomic fingerprinting에 의한 주요 식중독 그람 음성 세균 4속의 구별)

  • Jung, Hye-Jin;Park, Sung-Hee;Seo, Hyeon-A;Kim, Young-Joon;Cho, Joon-Il;Park, Sung-Soo;Song, Dae-Sik;Kim, Keun-Sung
    • Korean Journal of Food Science and Technology
    • /
    • v.37 no.6
    • /
    • pp.1005-1011
    • /
    • 2005
  • Widespread distributions of repetitive DNA elements in bacteria genomes are useful for analysis of genomes and should be exploited to differentiate food-borne pathogenic bacteria among and within species. Enterobacterial repetitive intergenic consensus (ERIC) sequence has been used for ERIC-PCR genomic fingerprinting to identify and differentiate bacterial strains from various environmental sources. ERIC-PCH genomic fingerprinting was applied to detect and differentiate four major Gram-negative food-borne bacterial pathogens, Esherichia coli, Salmonella, Shigella, and Vibrio. Target DNA fragments of pathogens were amplified by ERIC-PCR reactions. Dendrograms of subsequent PCR fingerprinting patterns for each strain were constructed, through which relative similarity coefficients or genetic distances between different strains were obtained numerically. Numerical comparisons revealed ERIC-PCR genotyping is effective for differentiation of strains among and within species of food-borne bacterial pathogens, showing ERIC-PCR fingerprinting methods can be utilized to differentiate isolates from outbreak and to determine their clonal relationships among outbreaks.

Molecular Identification of Pseudanabaena Strains and Analysis of 2-MIB Production Potential in the North Han River System (북한강 수역에 분포하는 Pseudanabaena 균주의 동정 및 2-MIB 생산 잠재성 분석)

  • Kim, Keonhee;Lee, Sejin;Seo, Kyunghwa;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
    • /
    • v.53 no.4
    • /
    • pp.344-354
    • /
    • 2020
  • Identification of the target species of 2-MIB (2-methyllisoborneol) production is crucial in the management of off-flavor problem in the freshwater system. This study was conducted to identify 2-MIB-producing Pseudanabaena strains occurring in the North Han River system using molecular genetic method. Eleven phenotypes of Pseudanabaena were isolated from several mainstream sites of the North Han River, including Sambong-ri, Joam-myun, and Lake Uiam areas. Despite of morphological similarity of the strains, the phylogenetic analysis using 16S rDNA classified them into different species with low genetic similarity (40~55%). Isolated Pseudanabaena strains were converged to four species; Pseudanabaena cinerea, P. yagii, P. mucicola, and P. redekei. Among them, the 2-MIB synthesizing gene (mibC) was detected in P. cinerea, P. yagii, and P. redekei. However, actual 2-MIB production was detected only in P. cinerea and P. redekei based on gas chromatography analysis. This study is the first report of the molecular identification of Pseudanabaena strains and their 2-MIB production potential in Korea. The results of this study provides an evidence of species diversity of Pseudanabaena occurring in the North Han River.

Evaluation of Genetic Safety in Genome-editing Rice Through Comparative Analysis of Genetic and Agronomic Traits (유전적 특성과 농업형질의 비교분석을 통한 유전자 교정 벼의 안전성 평가)

  • Seung-Kyo Jeong;Dohyeong Gwon;Bae-Hyeon Lee;Jeong-Hwan Suh;Rahmatullah Jan;Jae-Ryoung Park;Taehun Ryu;Kyung-Min Kim
    • Journal of Life Science
    • /
    • v.34 no.8
    • /
    • pp.567-575
    • /
    • 2024
  • New breeding techniques (NBT) recognize specific DNA sequences and remove, modify, or insert DNA at a desired location, and can be used to treat genetic diseases in humans or to improve the traits of livestock or crops. In this study, we conducted a comparative analysis of various agricultural traits and assessed the safety of gene transferability in third-generation genome-editing rice (OsCKq1-G3) with T and G nucleotide insertions developed using the CRISPR/Cas9 SDN-1 method, in comparison to its parental line (Oryza sativa L., cv Ilmi). The analyzed traits included heading date, culm length, panicle length, tiller number, yield, germination rate, viviparous germination rate, shattering, after wintering seed viability, the presence of toxins and allergens. The target trait, heading date, exhibited a high significant difference of approximately 5 days. Culm length, panicle length, tiller number, yield showed no significant differences compared to the parental line. No T-DNA bands indicating gene transfer were detected. In the third generation of genome-edited rice, the T-DNA was confirmed to be eliminated as successive generations advanced through self-pollination. Through the analysis of germination rate, viviparous germination rate, shattering, and after wintering viability, we confirmed that the genome-editing rice has no potential for weediness. The ORF and amino acid sequences of the genome-editing rice did not reveal any toxins and allergens. The results of this study can be utilized as important data for the environmental risk assessment of genome-editing rice.