• Title/Summary/Keyword: PCR-PP

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Composting and Fertilizing Characteristics of Poultry Manure Mixture with Compressed Expansion Rice Hull as Bulking Agent (수분조절제로서 팽화왕겨를 이용한 계분 발효 특성 및 시비 효과)

  • Kim, Young-Sun;Cho, Sung-Hyun;Lee, Hoon-Soo;Lee, Geung-Joo
    • Journal of the Korea Organic Resources Recycling Association
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    • v.28 no.4
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    • pp.5-13
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    • 2020
  • This study was conducted to evaluate effects of compressed expansion rice hull (CERH) as bulking agent on physicochemical properties of composting poultry manure (PM) and of its fertilization on lettuce and pak choi growth. Treatments were designed as follows; sawdust treatment (90% PM + 10% sawdust; SP), peatmoss treatment (90% PM + 10% peatmoss; PP), and CERH treatments [PCR1 (90% PM + 10% CERH 1.3 mm) and PCR2 (90% PM + 10% CERH 3.0 mm)]. Physicochemical properties such as temperature, water content, pH, and total carbon of composted poultry piles for 31 days were unaffected by various bulking agents. However, total nitrogen content in compost pile was higher in PP and PCR1 than that of SP or PCR2. After composting for 31 days, content ranges of N, P2O5, and K2O in the composting PM piles were 19.1~19.7%, 47.6~51.6%, 2.76~3.65%, and 2.53~2.90%, respectively. As compared to SP treatment, dry weight of lettuce treated with PP and PCR1 increased by more than 10%, but only in PP for pak choi. These results indicated that CERH 1.3 could be used as bulking agents for composting PM on behalf of peatmoss or sawdust.

Multiple Genetic Marker Analysis with Using Quantitative RT-PCR in Gastric Cancer (위암에서 정량적 역전사 중합효소연쇄반응을 이용한 다중 표지자 분석)

  • Yoo, Moon-Won;Lee, Hyuk-Joon;Choi, Soo-Min;Yu, Ji-Eun;Hur, Keun;Kim, Young-Kook;Yang, Han-Kwang
    • Journal of Gastric Cancer
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    • v.7 no.2
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    • pp.59-66
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    • 2007
  • Purpose: This study was aimed at evaluating the diagnostic validity of peritoneal dissemination of gastric cancer cells by performing multiple genetic marker analysis via quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) in gastric cancer cell lines and gastric cancer tissues. Materials and Methods: Quantitative RT-PCR was performed on 12 human gastric cancer cell lines and 10 gastric cancer tissues with four mRNAs of carcinoembryonic antigen (CEA), Cytokeratin 20 (CK20), dopa decarboxylase (DDC), and L-3-phosphoserine phosphatase (L3PP). Results: Out of the 12 human gastric cancer cell lines we tested, CEA was overexpressed in four cell lines (33%), CK20 in one (8%), DDC in six (50%) and L3PP was expessed in all the lines (100%). Out of the 10 gastric cancer tissues we tested, CEA was overexpressed in nine tissues, CK20 in eight, DOC in nine and L3PP was overexpressed in all the tissues. L3PP was overexpressed in all the gastric cancer cell lines and tissues, but the levels of overexpression were lower than those of CEA and DDC. Conclusion: Multiple genetic marker analysis can compensate for the weak points of single marker analysis when testing gastric cancer, and three mRNAs of CEA, DDC and L3PP can be used as candidate genes.

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Development of Multiplex PCR for Simultaneous Detection of Citrus Viruses and the Incidence of Citrus Viral Diseases in Late-Maturity Citrus Trees in Jeju Island

  • Hyun, Jae Wook;Hwang, Rok Yeon;Jung, Kyung Eun
    • The Plant Pathology Journal
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    • v.33 no.3
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    • pp.307-317
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    • 2017
  • Satsuma dwarf virus (SDV) or Citrus mosaic sadwavirus (CiMV) were not consistently detected in RTPCR assay with the primer sets based on gene of Japan isolates. SDV and CiMV isolates were distinctively divided into two groups based on phylogenetic analysis of PP2 gene cloned from 22 Korean isolates, and the Korean CiMV and SDV isolates shared 95.5-96.2% and 97.1-97.7% sequence identity with Japanese isolate, respectively. We developed PP2-1 primer set based on the PP2 gene sequence of Korean isolates to simultaneously and effectively detect SDV and CiMV. And CTLV-2013 and CTV-po primer sets were newly designed for detection of Citrus tatter leaf virus (CTLV) and Citrus tristeza virus (CTV), respectively. Using these primer sets, a new multiplex PCR assay was developed as a means to simultaneously detect 4 citrus viruses, CTV, CTLV, SDV, and CiMV. The degree of detection by the multiplex PCR were consistent with those of uniplex RT-PCR for detection of each of the viruses. Therefore, the new multiplex PCR provides an efficient method for detecting 4 citrus viruses, which will help diagnose many citrus plants at the same time. We verified that 35.2% and 72.1% of 775 trees in 155 orchards were infected with SDV or CiMV (SDV/CiMV) and CTV by the multiplex-PCR assay, respectively, and CTLV was not detected in any of the trees tested.

Effects of Heme Oxygenase-1 on VEGF Expression in Rheumatoid Arthritis (류마티스 관절염에서 HO-1에 의한 VEGF 발현 유도에 대한 연구)

  • Lee, Seung-Hoon;Kwan, Sang-Chul;Byun, Seung-Jae;Jang, Sung-Jo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.871-877
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    • 2008
  • Heme oxygenase-1 (HO-1), an inducible heme-degrading enzyme, is expressed by macrophages and endothelial cells in response to various stresses and mediators of inflammation. HO-1 has been recently implicated in regulation of angiogenesis via expression of VEGF. The purpose of this study was to determine the effects of HO-1 modulation on the collagen-induced arthritis (CIA) model and on angiogenesis via up- regulation of VEGF expression in human synovial fibroblast. DBA/1J mice were treated with an inhibitor of HO-1, tin protoporphyrin IX (SnPP), or with an inducer of HO-1, cobalt protoporphyrin IX (CoPP), from day 1 to day 35 after CIA induction. The clinical evolution of disease was monitored visually. At the end of the experiment, histopathologic changes were examined on the joints. VEGF expression in paws were measured by immunohistochemical stain. mRNA expression of HO-1 and VEGF stimulated with various concentration of $TNF-{\alpha}$, CoPP accessed on human synovial fibroblast by RT-PCR. Effects of pretreatment with SnPP on mRNA expression of HO-1 and VEGF in the presence of CoPP and $TNF-{\alpha}$ in synovial fibroblast was accessed by Real-time RT-PCR. Administration of cobalt protoporphyrin IX significantly induced the inflammatory response, with increased arthritis index and expression of VEGF in the paws of the arthritis models. Treatment with SnPP significantly reduced the severity of CIA through inhibition of joint inflammation and cartilage destruction. The expression of VEGF were also significantly reduced by SnPP treatment in the paw. CoPPIX as inducer of HO-1, increased HO-1 and VEGF expression dose dependently in synovial fibroblast. In contrast, inhibition of HO-1 activity by SnPPIX abrogated CoPPIX-induced HO-1 and VEGF production in synovial fibroblast. Stimulation with $TNF-{\alpha}$ increased HO-1 and VEGF expression itself and showed additive effect on HO-1 and VEGF expression when it treated with CoPP. When SnPP was treated with CoPP and $TNF-{\alpha}$, it abrogated the CoPP induced HO-1 and VEGF expression and also abrogated $TNF-{\alpha}$ induced HO-1 and VEGF expression in synovial fibroblast. The effects of HO-1 induction in rheumatoid arthritis results in aggravation of arthritis via up-regulation of VEGF. I concluded that inhibition of the expression or activity of HO-1 could be a therapeutic target of rheumatoid arthritis.

Paralytic Peptide Binding Protein (PP-BP) Gene Expression During Egg Diapause and Its Multi-Gene Organization in the Silkworm Bombyx mori

  • Sirigineedi, Sasibhushan;Murthy, Geetha N.;Rao, Guruprasada;Ponnuvel, Kangayam M.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.26 no.1
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    • pp.31-40
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    • 2013
  • Paralytic peptide binding proteins (PP-BP) are 30KP proteins that show similarity to ENF binding proteins. The ENF-BP act as active regulators of ENF peptides. ENF peptides are multifunctional insect cytokines. The comparison of gene expression in diapause induced and non-diapause eggs at different time intervals after oviposition showed an upregulation of PP at 18h as well as PP-BP at 12 and 18h after oviposition along with few other genes. The current study has been taken up to investigate the role of PP as well as PP-BP in diapause induction in polyvoltine silkworms and to study the multigene organization of PP-BP in the Bombyx mori genome. The tissue specific expression analysis revealed that, PP-BP is highly expressed in fat body followed by egg and brain while no expression was observed in midgut. The expression levels of PP and PP-BP in diapause and non-diapause eggs from 0h to 48h after oviposition, validated through realtime PCR revealed that PP is highly expressed at 18 and 24h while PP-BP expression is higher at 12 and 18h time intervals suggesting their possible role in diapause induction. The whole genome survey of the PP-BP paralogous sequences revealed a total of 46 B. mori PP-BP homologs that are classified into 3 categories viz., ENF-BP, Typical 30KPs and serine/threonine rich 30KPs. These paralogous sequences are distributed on chromosomes 7, 20, 22 and 24, all 30KP and S/T rich 30KP proteins are present in the same locus of chromosome 20.

Cloning, Over-expression, and Characterization of YjgA, a Novel ppGpp-binding Protein

  • Gnanasekaran, Gopalsamy;Pan, SangO;Jung, Wontae;Jeong, Kwangjoon;Jeong, Jae-Ho;Rhee, Joon Haeng;Choy, Hyon E.;Jung, Che-Hun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.8
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    • pp.2419-2424
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    • 2013
  • Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

Genetic Variability among Different Populations of Root Knot Nematodes Based on Their Encumbrance Response to Pasteuria Isolates Using PCR-RFLP

  • Kamran, Muhammad;Javed, Nazir;Ullah, Ihsan;Nazir, Shahid;Jamil, Shakra;Iqbal, Muhammad Zafar;Abbas, Huma;Khan, Sajid Aleem;Haq, Muhammad Ehetisham ul
    • The Plant Pathology Journal
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    • v.35 no.1
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    • pp.51-62
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    • 2019
  • A great variable response was observed when PP-3 and PP-J encumbered with 116 populations of root knot nematode (RKN) at two different temperatures ($25{\pm}2^{\circ}C$ and $30{\pm}2^{\circ}C$) and concentrations ($10^4$ and $10^5$ spores/ml). The PCR reaction amplified intergenic region between cytochrome oxidase subunit II gene (COII) and large subunit of rRNA gene (lrRNA) of the mitochondrial genome of different RKN species. The primer C2F3 and 1108 identified M. incognita with the highest frequency (52.6%) followed by M. javanica (36.8%) and M. arenaria (10.5%). The sizes of PCR products were 1.7 kb for M. incognita and M. javanica populations while populations of M. arenaria produced 1.1 kb fragment. The digestion with Hinf I yielded three different fragment length patterns on 1.5 % agarose gel. From current research it is concluded that intra-Meloidogyne genetic variability exist in RKN populations which have better encumbrance with P. penetrans.

Evaluation of JC and Cytomegalo Viruses in Glioblastoma Tissue

  • Afshar, Reza Malekpour;Mollaei, Hamid Reza;Zandi, Bahare;Iranpour, Maryam
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.11
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    • pp.4907-4911
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    • 2016
  • Glioblastoma multiforme (GBM) is the most aggressive of the gliomas, a collection of tumors arising from glia in the central nervous system. Possible associations between the human cytomegalovirus (HCMV) and the JC virus with GBM are now attracting interest. Our present aim was to investigate the prevalence of the two viruses in Iranian patients from Kerman's cities in the south of Iran. In addition, the expression rates of pp65, large T antigen and p53 proteins were assessed and their relation with GBM evaluated using reverse transcription real time PCR (rReal Time PCR). A total of 199 patients with GBM cancer were enrolled, with $mean{\pm}SD$ ages of $50.0{\pm}19.5$ and $50.7{\pm}19.6$ years for males and females, respectively. The P53 rate was dramatically low suggesting an aetiological role,. Large T antigen expression was found in JC positive samples, while the PP65 antigen was observed in patients positive for CMV and JC. HCMV products and JC virus with oncogenic potential may induce the development of various tumors including glioblastomas. The JC virus produces an early gene product, T-antigen, which has the ability to associate with and functionally inactivate well-studied tumor suppressor proteins including p53 and pRB.

Preferential Expression of IgA Isotype Switching-associated Transcripts in Mouse Intestinal Lymphoid Tissues (마우스 내장 림프조직에서 우세하게 발현되는 IgA Isotype Switching 관련 전사체의 분석)

  • Chae, Byung-Chul;Chun, Sung-Ki;Seo, Goo-Young;Kim, Hyun-A;Kim, Pyeung-Hyeun
    • IMMUNE NETWORK
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    • v.5 no.4
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    • pp.215-220
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    • 2005
  • Background: Transforming growth factor-${\beta}$ (TGF-${\beta}1$) directs class switch recombination (CSR) to IgA isotype, which is a predominant antibody in mucosal surfaces. Although IgA is preferentially committed in mucosal lymphoid tissues, it is not definitely established whether hallmarks of IgA CSR such as IgA germ-line transcripts (GLT ${\alpha}$), post-switch transcripts (PST ${\alpha}$) and circle transcripts (CT ${\alpha}$) are readily expressed in such tissues. Therefore, we compared the expression of these transcripts among mouse Peyer's patches (PP), mesenteric lymph nodes (MLN), and spleen. Methods: Levels of GLTs, PSTs and CTs were measured by RT-PCR in isolated PPs, MLNs and spleen cells. Results: GLT ${\alpha}$ and PST ${\alpha}$ were well expressed in PP and MLN cells but in spleen cells. Similar patterns were observed in the expression of GL ${\gamma}$2b and PST ${\gamma}$2b. On the other hand, these transcripts were only inducible in spleen cells upon stimulated with LPS and TGF-${\beta}1$. In addition, CT${\alpha}$ and CT${\gamma}$2b were detected in PP cells. Conclusion: PP B cells readily express IgA GLT, PST, and CT. Overall expression patterns of these transcripts were similar in MLN cells. Thus, these results suggest that microenvironment of PP and MLN influences spontaneous IgA CSR, which lacks in systemic lymphoid tissues such as spleen.

Verification of Recycled Plastic Pellets and Mucuna pruriens Extracts Correlation of Antioxidant Efficacy (재활용 플라스틱 펠렛과 벨벳콩(Mucuna pruriens) 추출물의 항산화 효능 상관관계 검증)

  • Dong-Geun Han;Min-jun Bae;GE-GE;Hyun-Jeong Kim;Se-Gie Kim;Bong-Jeon An
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.1
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    • pp.29-38
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    • 2023
  • In this study, the correlation between recycled plastic pellets and the antioxidant efficacy of the extract of Mucuna pruriens (M. pruriens) was verified. After immersing recycled plastic pellets in M. pruriens extracts, they were stored for 30 days under various temperature conditions, and then analyzed for DPPH radical and ABTS+ radical erasing ability, SOD-like activity, total phenol and flavonoid content. Compared to the existing efficacy, DPPH radical elimination showed less than 30% activity change in ECOTRIA and PCR-PP pellets, and the largest decrease in activity of 46.06% was confirmed in Grico pellets. As a result of measuring ABTS+ radical erasure ability, less than 5% of efficacy change was confirmed in all pellets. As a result of measuring SOD-like activity, all showed a decrease in activity of more than 30%, and as a result of measuring the total phenol and flavonoid content, all pellets showed a tendency to decrease compared to the existing content. Through this results, it was confirmed that the physiological activity of M. pruriens extract decreased and changed by recycled plastic pellets, and based on the results of this study, recycled plastic pellets are believed to affect the existing efficacy and activity stability of M. pruriens extract.