• Title/Summary/Keyword: conventional-PCR

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Acceleration of Biological Denitrification by Using Bioelectrochemical Reactor (생물전기화학반응기를 이용한 생물학적 탈질반응의 촉진)

  • Chun, Ji-Eun;Yu, Jae-Cheul;Park, Young-Hyun;Seon, Ji-Yun;Cho, Sun-Ja;Lee, Tae-Ho
    • Journal of Environmental Science International
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    • v.21 no.8
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    • pp.989-996
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    • 2012
  • Nitrate contamination of water environments can create serious problems such as eutrophication of rivers. Conventional biological processes for nitrate removal by heterotrophic denitrification often need additional organic substrates as carbon sources and electron donors. We tried to accelerate biological denitrification by using bioelectrochemical reactor (BER) in which electrode works as an electron donor. Denitrification activity of 8 environmental samples from various sediments, soils, groundwaters, and sludges were tested to establish an efficient enrichment culture for BER. The established enrichment culture from a soil sample showed stable denitrification activity without any nitrite accumulation. Microbial community analysis by using PCR-DGGE method revealed that dominant denitrifiers in the enrichment culture were Pantoea sp., Cronobacter sakazakii, and Castellaniella defragrans. Denitrification rate ($0.08kg/m^3{\cdot}day$) of the enrichment culture in BER with electrode poised at -0.5 V (vs Ag/AgCl) was higher than that ($2.1{\times}10^{-2}kg/m^3{\cdot}day$) of BER without any poised potential. This results suggested that biological denitrification would be improved by supplying potential throughout electrode in BER. Further research using BER without any organic substrate addition is needed to apply this system for bioremediation of water and wastewater contaminated by nitrate.

Molecular Identification of Haemadipsa rjukjuana (Hirudiniformes: Haemadipsidae) in Gageo Island, Korea

  • Won, Sohyun;Park, Bae Keun;Kim, Baek Jun;Kim, Hye Won;Kang, Jun Gu;Park, Tae Seo;Seo, Hong Yul;Eun, Ye;Kim, Ki Gyoung;Chae, Joon Seok
    • Parasites, Hosts and Diseases
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    • v.52 no.2
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    • pp.169-175
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    • 2014
  • There are 60 species of blood-feeding land leeches, 50 species belonging to the family Haemadipsidae and 10 species belonging to the family Xerobdellidae. Despite recent papers on the land leeches, their taxonomic identification is not fully understood, especially at a species level. In Korea, there have been no historical records of the terrestrial leeches, but recently an unrecorded blood-feeding land leech was discovered at Gageo-do (Island), Korea. Molecular analysis was used to identify the species of 29 leeches collected from Mt. Dock-Sil in Gageo-do. Conventional PCR was conducted using nuclear 18S rRNA and mitochondrial cytochrome c oxidase subunit 1 (CO1) genetic marker. The 18S rRNA sequences revealed that the leeches share 99.9% identity with Haemadipsa rjukjuana (inhabiting Taiwan), and the CO1 sequences revealed that the leeches are very close to H. rjukjuana (inhabiting Taiwan). The CO1 sequences were separated into 2 categories, 1 with 94.6% and the other with 94.3% similarity to the H. rjukjuana L00115A (inhabiting Taiwan). This new finding of the land leech is the first record in Korea. In addition, the north range of the distribution of the blood-feeding leech (Hirudiniformes: Haemadipisidae) should be reconsidered including Korea.

Effects of nanoscale ridge/groovepattern arrayed surface on in vitro differentiation of multi-potent pulp cells derived from human supernumerary teeth

  • Kim, Daehwan;Jo, Hwansung;Lee, Jingu;Kim, Keesung;Roh, Sangho
    • International Journal of Oral Biology
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    • v.38 no.4
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    • pp.161-167
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    • 2013
  • Human dental pulp stem cells (DPSCs) are multi-potent mesenchymal stem cells that have several differentiation potentials. An understanding of thetissues that differentiate from these cells can provide insights for future regenerative therapeutics and tissue engineering strategies. The mesiodens is the most frequent form of supernumerary tooth from which DPSCs can differentiate into several lineages similar to cells from normal deciduous teeth. Recently, it has been shown that nanoscale structures can affect stem cell differentiation. In our presentstudy, we investigated the effects of a 250-nm nanoscale ridge/groove pattern array on the osteogenic and adipogenic differentiation of dental pulp cells from mesiodenscontaining human DPSCs. To this end, the expression of lineage specific markers after differentiation induction was analyzed by lineage specific staining and RT-PCR. The nanoscale pattern arrayed surface showed apositive effect on the adipogenic differentiation of DPSCs. There was no difference between nanoscale pattern arrayed surface and conventional surface groups onosteogenic differentiation. In conclusion, the nanoscale ridge/groove pattern arrayed surface can be used to enhance the adipogenic differentiation of DPSCs derived from mesiodens. This finding provides an improved understanding of the effects of topography on cell differentiation as well as the potential use of supernumerary tooth in regenerative dental medicine.

Cytogenetic Profile of De Novo B lineage Acute Lymphoblastic Leukemia: Determination of Frequency, Distribution Pattern and Identification of Rare and Novel Chromosomal Aberrations in Indian Patients

  • Bhandari, Prerana;Ahmad, Firoz;Dalvi, Rupa;Koppaka, Neeraja;Kokate, Prajakta;Das, Bibhu Ranjan;Mandava, Swarna
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7219-7229
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    • 2015
  • Background: Chromosomal aberrations identified in acute lymphoblastic leukemia (ALL) have an important role in disease diagnosis, prognosis and management. Information on karyotype and associated clinical parameters are essential to physicians for planning cancer control interventions in different geographical regions. Materials and Methods: In this study, we present the overall frequency and distribution patterns of chromosomal aberrations in both children and adult de novo B lineage ALL Indian patients using conventional cytogenetics, interphase FISH and multiplex RT-PCR. Results: Among the 215 subjects, cytogenetic results were achieved in 172 (80%) patients; normal karyotype represented 37.2% and abnormal 62.8% with a distribution as follows: 15.3% hypodiploidy; 10.3% hyperdiploidy; 15.8% t(9;22); 9.8% t(1;19); 3.7% t(12;21); 2.8% t(4;11); 2.8% complex karyotypes. Apart from these, we observed several novel, rare and common chromosomal rearrangements. Also, FISH studies using LSI extra-signal dual-color probes revealed additional structural or numerical changes. Conclusions: These results demonstrate cytogenetic heterogeneity of ALL and confirm that the incidence of chromosomal abnormalities varies considerably. To the best of our knowledge, this is one of the largest reported series of cytogenetic investigations in Indian B-lineage ALL cases. In addition, ongoing cytogenetic studies are warranted in larger groups of B-lineage ALL cases to identify newly acquired chromosomal abnormalities that may contribute to disease diagnosis and management.

Possibility in identifying species composition of fish communities using the environmental DNA metabarcoding technique - with the preliminary results at urban ecological streams (환경DNA 메타바코딩 기술을 활용한 수생태계 어류종 군집조사의 가능성 - 도시 생태하천 초기분석 자료를 중심으로)

  • Song, Young-Keun;Kim, Jong-Hee;Won, Su-Yeon;Park, Chan
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.22 no.6
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    • pp.125-138
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    • 2019
  • This study aims to highlight the possibility in identifying species composition of fish communities using the environmental DNA (eDNA) metabarcoding technique, from both of the technical introduction and the pilot test at urban ecological streams. This new emerging survey technique using eDNA is getting popular in the world as a compensating way for the conventional field survey. However, the application to the domestic cases has yet to be studied. We attempted to use this technique for identifying fish species observed at four survey points in Hwangguji-chon, Suwon City. As a result, the detected number of species by eDNA sampled once in May was significantly matched with the total number of observed species in annual field surveys. Additionally eDNA results indicated the presence possibility of the unobserved species in field last year, even though the validation may be required. This survey technique seems to be more efficient and applicable to diverse situations of the fields and species, thereby needs to be studied further. We discussed the pros and cons of the application and summarized the research directions in future.

Virological Response to Conventional Interferon Therapy Combined with Ribavirin against Various HCV Genotypes in Khyber Pakhtunkhwa, Pakistan

  • Ali, Sajid;Ahmad, Bashir;Ali, Ijaz;Mahmood, Nourin;Anwar, Naveed;Saeedi, Ilyas;Afridi, Jehan Zeb
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.5
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    • pp.2407-2410
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    • 2016
  • Background: Response to antiviral therapy has been linked to different genotypes and this impacts on clinical management. Data on general responses to standard interferon (IFN) against HCV infection exists but little is known regarding HCV genotype specific responses. Purpose: Therefore, we attempted to determine genotype specific responses of chronic HCV patients, having different HCV genotypes, to standard IFN and ribavirin combination therapy administered for a period of six months in Kybher Pakhtunkhwa province of Pakistan. Materials and Methods: HCV genotype was determined for all serum samples. Each patient received standard IFN combination therapy with ribavirin administered at dose of 3 MIU three times a week and 800-1200 mg/day, respectively, for a period of six months. After completion of this therapy, PCR was performed for all course completed subjects. Results: Out of total 51 selected patients the most abundant genotypes were 3a (49.0%) and 1a (21.6%) followed by 3b (9.8%), 1b (7.84%), 2a (7.84%) and untypable (3.94%). Moast responsive genotypes were 2a followed by 3a, with end of treatment responses of 77.7%, and 72.2%. Responses for 3b, 1b and untypable were 66.7%, 33.3% and 0%, respectively. Conclusions: IFN response is efficient in case of 2a and 3a genotypes while in case of untypable genotypes, further categorization is required to know about genomic sequences and to adopt some new regimes against these genotypes.

Genetically Modified Human Embryonic Stem Cells Expressing Nurr1 and Their Differentiation into Tyrosine Hydroxylase Positive Cells in vitro.

  • Cho, Hwang-Yoon;Lee, Chang-Hyun;Kil, Kwang-Soo;Yoon, Ji-Yeon;Shin, Hyun-Ah;Lee, Gun-Soup;Lee, Young-Jae;Kim, Eun-Young;Park, SePill;Lim, Jin-Ho
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.104-104
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    • 2003
  • As an effort to direct differentiation of human embryonic stem (hES, MB03) cells to dopamine-producing neuronal cells, Nurr1 was transfected using conventional transfection protocol into MB03 and examined the expression of tyrosine hydroylase (TH) after differentiation induced by retinoic acid (RA) and ascorbic acid (AA). Experimentally, cells were transfected with linearized Nurr1 cDNA in pcDNA3.1 (+)-hygovernight followed by selection in medium containing hygromycin-B (150 $\mu$/ml). Expression of Nurr1 mRNA was confirmed by RT-PCR and protein by immunocytochemistry in the drug resistant clones. In order to study the effect of Nurr1 protein on the differentiation pattern of ES cells, one of the positive clones (MBNr24) was allowed to form embryoid body (EB) for 2 days and were induced to differentiate for another 4 days using RA (1 $\mu M$) and AA (50 mM) (2-/4+ protocol) followed by selection in N2 medium for 10 or 20 days. After 10 days in N2 medium, cells immunoreactive to anti-GFAP, anti-TH, or anti-NF200 antibodies were 38.8%, 11%, and 20.5%, respectively. After 20 days in N2 medium, cells expressing GFAP, TH, or NF200 were 28%, 15% and 44.8%, respectively but approximately 9% of MB03 expressed TH protein when the cells were induced to differentiate using a similar prorocol, These results suggest that ectopic expression of Nurr1 enhances generation of TH+ cells as well as neuronal cells when hES cells were differentiated by 2-/4+ protocol.

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Inhibition of adhesion and biofilm formation in Escherichia coli O157:H7 by diosmin (다이오스민(diosmin)에 의한 병원성 대장균 세포부착 및 생물막 형성 억제)

  • Kim, Hyun Jung;Kim, Seung Min
    • Korean Journal of Food Science and Technology
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    • v.50 no.4
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    • pp.414-419
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    • 2018
  • Escherichia coli O157:H7 is one of the most common foodborne pathogens responsible for outbreaks of hemorrhagic colitis, which can lead to the life-threatening hemolytic-uremic syndrome. In this study, we identified phytochemicals that specifically inhibit the expression of LEE operon in E. coli O157:H7. Among phytochemicals, diosmin decreased the adherence of E. coli O157:H7 towards Caco-2 cells in vitro (p<0.01) and its biofilm formation activity (p<0.05). Quantitative RT-PCR analysis revealed that the transcripts of Ler-regulated genes and genes related to curli production were significantly reduced in the presence of diosmin. However, diosmin does not affect bacterial viability, indicating that the resistance rate to diosmin was remarkably low. Overall, these results provide significant insights into the development of a novel anti-infective agent that is different from conventional antibiotics.

Overexpression of Anti-apoptotic Molecules and Sax Translocation to Mitochondria by Pharbitis Nil Extracts in AGS

  • Ko Seong-Gyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1843-1849
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    • 2004
  • Conventional medicines have usually sorted to a number of treatments such asoperation, radiotherapy, and chemotherapy. The existing anti-cancer agents, designed to eradicate cancer cells, have strong toxicities, also with leading to harmful side effects. Recently, a number of researches on natural products have been actively carried out in efforts to develop new treatments that can decrease side effects or increase anti-cancer effects. We performed this study to understand the molecular basis underlying the antitumor effects of Pharbitis nil, and Plantago asiatica, which have been used for herbal medicinal treatments against cancers in East Asia. We analyzed the effects of these medicinal herbs on proliferation and on expression of cell growth/apoptosis related molecules, with using an AGS gastric cancer cell line. The treatment of Pharbitis nil dramatically reduced cell viabilities in a dose and time-dependent manner, but Plantago asiatica didn't. FACS analysis and Annexin V staining assay also showed that Pharbitis nil induce apoptotic cell death of AGS. Expression analyses via RT-PCR and Western blots revealed that Pharbitis nil didn't increase expression of the p53 and its downstream effector p21/sup wafl/, and that the both increased expression of apoptosis related Sax and cleavage of active caspase-3 protein. We also confirmed the translocation of Sax to mitochondria. Collectively, our data demonstrate that Pharbitis nilinduce growth inhibition and apoptosis of human gastric cancer cells, and these effects are correlated with down- and up-regulation of growth-regulating apoptotic and tumor suppressor genes, respectively.

Development and Evaluation of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Tylenchulus semipenetrans Using DNA Extracted from Soil

  • Song, Zhi-Qiang;Cheng, Ju-E;Cheng, Fei-Xue;Zhang, De-Yong;Liu, Yong
    • The Plant Pathology Journal
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    • v.33 no.2
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    • pp.184-192
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    • 2017
  • Tylenchulus semipenetrans is an important and widespread plant-parasitic nematode of citrus worldwide and can cause citrus slow decline disease leading to significant reduction in tree growth and yield. Rapid and accurate detection of T. semipenetrans in soil is important for the disease forecasting and management. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed to detect T. semipenetrans using DNA extracted from soil. A set of five primers was designed from the internal transcribed spacer region (ITS1) of rDNA, and was highly specific to T. semipenetrans. The LAMP reaction was performed at $63^{\circ}C$ for 60 min. The LAMP product was visualized directly in one reaction tube by adding SYBR Green I. The detection limit of the LAMP assay was $10^{-2}J2/0.5g$ of soil, which was 10 times more sensitive than conventional PCR ($10^{-1}J2/0.5g$ of soil). Examination of 24 field soil samples revealed that the LAMP assay was applicable to a range of soils infested naturally with T. semipenetrans, and the total assay time was less than 2.5 h. These results indicated that the developed LAMP assay is a simple, rapid, sensitive, specific and accurate technique for detection of T. semipenetrans in field soil, and contributes to the effective management of citrus slow decline disease.