• Title/Summary/Keyword: replication control

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Bacterial community structure of paddy fields as influenced by heavy metal contamination

  • Tipayno, Sherlyn;Samaddar, Sandipan;Chatterjee, Poulami;Halim, MD Abdul;Sa, Tongmin
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.245-245
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    • 2017
  • Heavy metal pollution of agricultural soils affects land productivity and has impact on the quality of surrounding ecosystem. Soil microbial community parameters are used as reliable indices for assessing quality of agricultural lands under metal stress. This study investigated bacterial community structure of polluted and undisturbed paddy soils to elucidate soil factors that are related to alteration of bacterial communities under conditions of metal pollution. No obvious differences in the richness or diversity of bacterial communities were observed between samples from polluted and control areas. The bacterial communities of three locations were distinct from one another, and each location possessed distinctive set of bacterial phylotypes. The abundances of several phyla and genera differed significantly between study locations. Variation of bacterial community was mostly related to soil general properties at phylum level while at finer taxonomic levels concentrations of arsenic and lead were significant factors. According to results of bacterial community functional prediction, the soil bacterial communities of metal polluted locations were characterized by more abundant DNA replication and repair, translation, transcription and nucleotide metabolism pathway enzymes while amino acid and lipid metabolism as well as xenobiotic biodegradation potential was reduced.Our results suggest that the soil microbial communities had adapted to the elevated metal concentrations in the polluted soils as evidenced by changes in relative abundances of particular groups of microorganisms at different taxonomic resolution levels, and by altered functional potential of the microbial communities.

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A Homeotic Gene, Hoxc8, Regulates the Expression of Proliferating Cell Nuclear Antigen in NIH3T3 Cell

  • Min, Hye-Hyun;Kang, Myeng-Mo;Kim, Myoung-Hee
    • Biomedical Science Letters
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    • v.13 no.3
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    • pp.239-244
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    • 2007
  • Hoxc8 is one of the homeotic developmental control genes regulating the expression of many downstream target genes, through which animal body pattern is established during embryonic development. In previous proteomics analysis, proliferating cell nuclear antigen (PCNA) which is also known as cyclin, has been implied to be regulated by Hoxc8 in F9 murine embryonic teratocarcinoma cell. When the 5' upstream region of PCNA was analyzed, it turned out to contain 20 Hox core binding sites (ATTA) in about 1.17 kbp (kilo base pairs) region ($-520{\sim}-1690$). In order to test whether this region is responsible for Hoxc8 regulation, the upstream 2.3 kbp fragment of PCNA was amplified through PCR and then cloned into the pGL3 basic vector containing a luciferase gene as a reporter. When the luciferase activity was measured in the presence of effector plasmid (pcDNA : c8) expressing murine Hoxc8, the PCNA promoter driven reporter activity was reduced. To confirm whether this reduction is due to the Hoxc8 protein, the siRNA against Hoxc8 (5'-GUA UCA GAC CUU GGA ACU A-3' and 5'-UAG UUC CAA GGU CUG AUA C-3') was prepared. Interestingly enough, siRNA treatment up regulated the luciferase activity which was down regulated by Hoxc8, indicating that Hoxc8 indeed regulates the expression of PCNA, in particular, down regulation in NIN3T3 cells. These results altogether indicate that Hoxc8 might orchestrate the pattern formation by regulating PCNA which is one of the important proteins involved in several processes such as DNA replication and methylation, chromatin remodeling, cell cycle regulation, differentiation, as well as programmed cell death.

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In Vivo Excision and Amplification of Large Human Genomic Segments Using Cre/loxP-and EBNA-1/oriP-mediated Machinery

  • Yoon, Young-Geol;Choi, Ja-Young;Kim, Jung-Min;Lee, Jun-Hyoung;Kim, Sun-Chang
    • BMB Reports
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    • v.34 no.4
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    • pp.322-328
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    • 2001
  • Excision and amplification of pre-determined, large genomic segments (taken directly from the genome of a natural host, which provides an alternative to conventional cloning in foreign vectors and hosts) was explored in human cells. In this approach, we devised a procedure for excising a large segment of human genomic DNA, the iNOS gene, by using the Cre/loxP system of bacteriophage P1 and amplifying the excised circles with the EBNA-1/oriP system of the Epstein-Barr virus. Two loxP sequences, each of which serves as a recognition site for recombinase Cre, were integrated unidirectionally into the 5'-UTR and 3'-UTR regions of the iNOS gene, together with an oriP sequence for conditional replication. The traps-acting genes cre and EBNA-1, which were under the control of a tetracycline responsive $P_{hcmv^*-1}$ promoter, were also inserted into the 5'-UTR and 3'-UTR regions of the iNOS gene, respectively, by homologous recombination. The strain carrying the inserted elements was stably maintained until the excision and amplification functions were triggered by the induction of cre and EBNA-1. Upon induction by doxycycline, Cre excised the iNOS gene that was flanked by two ZoxP sites and circularized it. The circularized iNOS gene was then amplified by the EBNA-1/oriP-system. With this procedure, approximately a 45.8-kb iNOS genomic fragment of human chromosome 17 was excised and successfully amplified in human cells. Our procedure can be used effectively for the sequencing of unclonable genes, the functional analysis of unknown genes, and gene therapy.

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Development of Synthetic Jet Micro Air Pump (Synthetic Jet 마이크로 에어펌프의 개발)

  • Choi, J.P.;Kim, K.S.;Seo, Y.H.;Ku, B.S.;Jang, J.H.;Kim, B.H.
    • Transactions of Materials Processing
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    • v.17 no.8
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    • pp.594-599
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    • 2008
  • This paper presents a micro air pump based on the synthetic jet to supply reactant at the cathode side for micro fuel cells. The synthetic jet is a zero mass flux device that converts electrical energy into the momentum. The synthetic jet actuation is usually generated by a traditional PZT-driven actuator, which consists of a small cylindrical cavity, orifices and PZT diaphragms. Therefore, it is very important that the design parameters are optimized because of the simple configuration. To design the synthetic jet micro air pump, a numerical analysis has been conducted for flow characteristics with respect to various geometries. From results of numerical analysis, the micro air pump has been fabricated by the PDMS replication process. The most important design factors of the micro air pump in micro fuel cells are the small size and low power consumption. To satisfy the design targets, we used SP4423 micro chip that is high voltage output DC-AC converter to control the PZT. The SP4423 micro chips can operate from $2.2{\sim}6V$ power supply(or battery) and is capable of supplying up to 200V signals. So it is possible to make small size controller and low power consumption under 0.1W. The size of micro air pump was $16{\times}13{\times}3mm^3$ and the performance test was conducted. With a voltage of 3V at 800Hz, the air pump's flow rate was 2.4cc/min and its power consumption was only 0.15W.

Preliminary assessment of correlation between T-lymphocyte responses and control of porcine reproductive and respiratory syndrome virus (PRRSV) in piglets born after in-utero infection of a type 2 PRRSV

  • Cha, Sang-Ho;Bandaranayaka-Mudiyanselage, Carey;Bandaranayaka-Mudiyanselage, Chandima B.;Ajiththos, Dharani;Yoon, Kyoung-Jin;Gibson, Kathleen A.;Yu, Ji-Eun;Cho, In-Soo;Lee, Stephen S.;Chung, Chungwon J.
    • Korean Journal of Veterinary Research
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    • v.58 no.1
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    • pp.9-16
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    • 2018
  • A preliminary study into the protective mechanisms of adaptive immunity against porcine reproductive and respiratory syndrome virus (PRRSV) in piglets (n = 9) born to a gilt challenged intranasally with a type-2 PRRSV. Immune parameters (neutralizing antibodies, $CD3^+CD4^+$, $CD3^+CD8^+$, $CD3^+CD4^+CD8^+$ T-lymphocytes, and PRRSV-specific interferon $(IFN)-{\gamma}$ secreting T-lymphocytes) were compared with infection parameters (macro- and microscopic lung lesion, and PRRSV-infected porcine alveolar macrophages ($CD172{\alpha}^+PRRSV-N^+\;PAM$) as well as with plasma and lymphoid tissue viral loads. Percentages of three T-lymphocyte phenotypes in 14-days post-birth (dpb) peripheral blood mononuclear cell (PBMC) had significant negative correlations with percentages of $CD172{\alpha}^+PRRSV-N^+\;PAM$ (p < 0.05) as well as with macroscopic lung lesion (p < 0.01). Plasma and tissue viral loads had significant (p < 0.05) negative correlations with $CD3^+CD4^+CD8^+$ T-lymphocyte percentage in PBMC. Frequencies of $CD3^+CD8^+$ and $CD3^+CD4^+$ T-lymphocytes in 14-dpb PBMC had significant negative correlations with of lymph node (p = 0.04) and lung (p = 0.002) viral loads. $IFN-{\gamma}$-secreting T-lymphocytes frequency had a significant negative correlation with gross lung lesion severity (p = 0.002). However, neutralizing antibody titers had no significant negative correlation (p > 0.1) with infection parameters. The results indicate that T-lymphocytes contribute to controlling PRRSV replication in young piglets born after in-utero infection.

Modulation of Chemical Carcinogen-Induced Unscheduled DNA Synthesis by Dehydroepiandrosterone (DHEA) in the Primary Rat Hepatocytes

  • Kim, Seung-Hee;Han, Hyung-Mee;Kang, Seog-Youn;Jung, Ki-Kyung;Kim, Tae-Gyun;Oh, Hye-Young;Lee, Young-Kyung;Rheu, Hang-Mook
    • Archives of Pharmacal Research
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    • v.22 no.5
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    • pp.474-478
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    • 1999
  • Modulation of unscheduled DNA synthesis by dehydroepiandrosterone (DHEA) after exposure to various chemical carcinogens was investigated in the primary rat hepatocytes. Unscheduled DNA synthesis was induced by treatment of such direct acting carcinogens as methly methanesulfonate (MMS) and ethyl methanesulfonate (EMS) or procarcinogens including benzo(a)pyrene (BaP) and 7, 12-dimethylbenz(a)anthracene (DMBA). Unscheduled DNA synthesis was determined by measuring [methyl-3H]thymidine radioactivity incorporated into nuclear DNA of hepatocytes treated with carcinogens in the presence or absence of DHEA. Hydroxyurea $(5{\times}10^{-3} M)$was added to growth medium to selectively suppress normal replication. DHEA at concentrations ranging from $(1{\times}10^{-6} M)$ to$(5{\times}10^{-4} M)$ did not significantly inhibit unscheduled DNA synthesis induced by either MMS $(1{\times}10^{-4} M)$ or EMS $(1{\times}10^{-2} M)$. In contrast, DHEA-significantly inhibited unscheduled DNA synthesis induced by BaP $(6.5{\times}10^{-5} M)$ and DMBA.$(2{\times}10^{-5} M)$. DHEA-induced hepatotoxicity in rats was examined using lactate dehydrogenase (LDH) release as an indicator of cytotoxicity. DHEA exhibit no significant increase in LDH release compared with the control at 18 h. These data suggest that nontoxic concentration of DHEA does not affect the DNA excision repair process, but it probably influence the enzymatic system responsible for the metabolic activation of procarcinogens and thereby decreases the amount of the effective DNA adducts formed by the ultimate reactive carcinogenic species.

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Control of X Chromosome Reactivation and Determination of the Ratio of Sex Chromosome to Autosome in Embryonal Carcinoma Cell-Somatic Cell Hybrids (배종양 세포와 체세포 간의 융합 세포에서 X 염색체 재활성화의 조절과 성염색체에 대한 상염색체 비율의 결정)

  • 이광호
    • The Korean Journal of Zoology
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    • v.39 no.1
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    • pp.75-88
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    • 1996
  • OTF9-63 (OTF9) and P19S1O1A1 (P19) embryonal carcinoma (EC) cells were examined for their ability to produce the readivation of inactive X chromosomes from somatic cells. They were hybridized with various somatic cells and resulting HATr EC-somatic cell clones were analysed for their morphology, chromosomal replication pafterns and expression proffies of X-linked and distantiy located genes, Hprt and Pgk-1. The results demonstrated that 0RF9 cells could reactivate the inactive X chromosome whereas P19 cells could not. In adition, EC-somatic cell hybrids tended to reduce the number of sex chromosomes in long-term culture, resulting m 1:2 ratio of sex chromosomes to autosomes The use of EC cell hybrids provides an experimental system for studying the mechanism(s) of the X-reactivatio that is initiated and maintained from meiotic prophase of oogenesis to early embryogenesis.

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MITOCHONDRIAL DNA DELETION AND IMPAIRMENT OF MITOCHONDRIAL BIOGENESIS ARE MEDIATED BY REACTIVE OXYGEN SPECIES IN IONIZING RADIATION-INDUCED PREMATURE SENESCENCE

  • Eom, Hyeon-Soo;Jung, U-Hee;Jo, Sung-Kee;Kim, Young-Sang
    • Journal of Radiation Protection and Research
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    • v.36 no.3
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    • pp.119-126
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    • 2011
  • Mitochondrial DNA (mtDNA) deletion is a well-known marker for oxidative stress and aging, and contributes to harmful effects in cultured cells and animal tissues. mtDNA biogenesis genes (NRF-1, TFAM) are essential for the maintenance of mtDNA, as well as the transcription and replication of mitochondrial genomes. Considering that oxidative stress is known to affect mitochondrial biogenesis, we hypothesized that ionizing radiation (IR)-induced reactive oxygen species (ROS) causes mtDNA deletion by modulating the mitochondrial biogenesis, thereby leading to cellular senescence. Therefore, we examined the effects of IR on ROS levels, cellular senescence, mitochondrial biogenesis, and mtDNA deletion in IMR-90 human lung fibroblast cells. Young IMR-90 cells at population doubling (PD) 39 were irradiated at 4 or 8 Gy. Old cells at PD55, and H2O2-treated young cells at PD 39, were compared as a positive control. The IR increased the intracellular ROS level, senescence-associated ${\beta}$-galactosidase (SA-${\beta}$-gal) activity, and mtDNA common deletion (4977 bp), and it decreased the mRNA expression of NRF-1 and TFAM in IMR-90 cells. Similar results were also observed in old cells (PD 55) and $H_2O_2$-treated young cells. To confirm that a increase in ROS level is essential for mtDNA deletion and changes of mitochondrial biogenesis in irradiated cells, the effects of N-acetylcysteine (NAC) were examined. In irradiated and $H_2O_2$-treated cells, 5 mM NAC significantly attenuated the increases of ROS, mtDNA deletion, and SA-${\beta}$-gal activity, and recovered from decreased expressions of NRF-1 and TFAM mRNA. These results suggest that ROS is a key cause of IR-induced mtDNA deletion, and the suppression of the mitochondrial biogenesis gene may mediate this process.

Influence of Compost Recycling and Magnesium Supplement on Physical and Chemical Traits of Animal Manure Compost

  • Lee, Jin-Eui;Kwag, Jung-Hoon;Ra, Chang-Six
    • Journal of Animal Science and Technology
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    • v.52 no.6
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    • pp.513-519
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    • 2010
  • A series of experiments were performed to study the influence of the following parameters on the physical traits and composition of swine manure compost: (1) addition of magnesium (Mg) at a molar ratio of 1.2 with respect to $PO_4$, and (2) reutilization of compost containing $MgNH_4PO_4{\cdot}6H_2O$ (magnesium ammonium phosphate, MAP). Three independent batch tests were conducted for replication: batch test I-control (C) and Mg added (T), batch test II-C, T and compost recycle ($T_{R1}$), and batch test III-C, T and compost recycle ($T_{R2}$). Magnesium addition and compost reutilization had no adverse effect on the degradation of organic matter. Reuse of the compost, however, had a clear effect on the total nitrogen (TN) and total phosphorus (TP) contents in the final compost. Repeated compost reutilization as a bulking material was resulted in composts rich in N and P. Upon adding the Mg supplement to the composting materials, the ortho-phosphate (OP) to TP ratio decreased due to the MAP crystallization reaction. The decrease in the OP/TP ratio and the increase in the TP content of the compost indicate that water-soluble phosphate is converted into a slow-release phosphate by the formation of crystals during composting. X-ray diffraction analysis of the irregular shaped crystals in the compost indicated that they are MAP crystals and that the crystallization of MAP begins immediately after the addition of the Mg supplement. The Mg addition to composting materials and the reutilization of compost as a bulking material would be a practical means to conserve nutrient content.

Evaluating the Effects of Salicornia Extract on Performance, Egg Quality and Blood Profile of Laying Hens

  • Mohammadi, Mohsen;Li, Hanlin;Kim, In Ho
    • Korean Journal of Poultry Science
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    • v.42 no.3
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    • pp.191-196
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    • 2015
  • The current experiment was conducted to evaluate the effects of adding Salicornia extract to the drinking water on the performance, egg quality, and blood profile of laying hens. A total of 216 Hy-Line Brown laying hens at 40 weeks of age were used in a 10-week experiment. The birds were allotted into three experimental treatments with three replications per treatment and 24 birds per replication. The treatments were CON (basal diet), T1 (1 cc of Salicornia extract per liter of drinking water), and T2 (5 cc of Salicornia extract per liter of drinking water). The collected data were analyzed using the SAS package program. The results indicated that addition of Salicornia extract to the drinking water of laying hens did not cause any negative effects on the performance, egg quality, or blood profile. Compared to the control treatment, the treatments with Salicornia extract remarkably increased egg production (P<0.05) in the last week of the study, improved egg shell thickness and significantly reduced the egg breaking rate (P<0.05). The results of this study showed that the addition of Salicornia extract improved egg shell quality; thus, Salicornia extract can decrease the egg breaking rate and increase production on commercial farms.