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Analysis of Gene Expression in Helicobacter pylori-associated Nodular Gastritis in Children Using Microarray (소아의 Helicobacter pylori 감염에 의한 결절성 위염의 유전자 발현 양상 분석)

  • Yang, Hye-Ran;Ko, Jae-Sung;Seo, Jeong-Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.13 no.1
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    • pp.7-22
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    • 2010
  • Purpose: Nodular gastritis is a characteristic finding of Helicobacter pylori infection in children. The aim of this study was to evaluate the difference in gene expression in the gastric mucosa of H. pylori-infected and non-infected children, and to analyze the difference in gene expression using cDNA microarray analysis of nodular gastritis caused by H. pylori infection. Methods: Twelve children (6 boys and 6 girls; mean age 9.8 years) who underwent upper gastrointestinal endoscopy and biopsy at Seoul National University Bundang Hospital were included in the study. The subjects were divided into three groups according to the presence of H. pylori infection and nodular gastritis on endoscopic examination. Gastric mucosa tissue was kept at $-70^{\circ}C$ and RNA was extracted to perform cDNA microarray analysis in each patient. Results: cDNA microarray analysis in children revealed a clear distinction between H. pylori-infected and non-infected gastric mucosa. Specifically, 182 over-expressed genes and 29 under-expressed genes were identified in H. pylori-infected gastric mucosa compared to non-infected mucosa. H. pylori-infected nodular gastritis revealed different gene expression patterns from H. pylori-infected normal gastric mucosa; five genes were over-expressed and five genes were under-expressed. Conclusion: In the presence of H. pylori infection, gastric mucosa shows distinct differences in gene expression, and nodular gastritis with H. pylori infection in children may be associated with over- or under-expression of some genes. Further studies are required to clarify the host response and the pathogenesis of nodular gastritis in children.

Genotyping and Clinical study of Mycoplasma pneumoniae During 1996-1997 in Korea (1996년과 1997년 소아에서 유행한 Mycoplasma pneumoniae의 Genotype에 따른 임상적 연구)

  • Lim, Hong Hee;Ahn, Byung Moon;Kim, Eun Ryoung;Choi, Sug Ho;Moon, Young Ho;Kim, Il Soo
    • Pediatric Infection and Vaccine
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    • v.5 no.1
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    • pp.79-87
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    • 1998
  • Purpose : The P1 protein of Mycoplasma pneumoniae mediates the attachment of the pathogen to its host cell and elicits a strong humoral immune response during infection with this organism. Mycoplasma pneumoniae strains can be classified into two groups(I and II) by PCR method of P1 cytadhesin gene. In this study, we evaluated the prevalence, epidemiological and clinical characteristics of each group. Methods : From 155 patients with Mycoplasma pneumoniae, who admitted to the Department of Pediatrics, Sung-Ae and Kwangmyung Sung-Ae Hospital between November 1996 and October 1997, we collected their throat swabs or nasopharyngeal aspirates for DNA extraction and serum for indirect hemagglutination test of Mycoplasma pneumoniae. The group specific PCR amplification were performed using specific oligonucleotide primers designed for P1 gene genotyping. Results : Group I(137 patients, 88.4%) occurred frequently than group II(18 patients, 11.6%). In both group, the most prevalent season was winter in 1996(Nov. to Dec.) and fall in 1997(Aug. to Oct.) The prevalent age was four to six years old. The number of male was more than female in both group; Group 1(1.2:1), Goup 2(1.6:1). No significant relationship were found between two groups in duration of fever and hospital days(P>0.05). The rate of high antibody titers(>1:5120) was lower in group I(6/137, 4.4%) than group II(2/18, 11.1%). Conclusion : Group I was much more prevalent than group II during 1996~1997 in Korea. There was no difference between two groups in epidemiological and clinical parameters except the rate of high antibody titers. Further follow-up survey will be needed for the epidemiologic and clinical studies of Mycoplasma pneumoniae in Korea.

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Regulation of Gb3 Expression on Dendritic Cells (수지상세포에 있어서 베로독소 수용체의 발현조절)

  • Lim, Suk-Hwan;Kim, Gi-Young;Kim, Hyung-Chun;Kim, Young-Hee;Son, Yong-Hae;Oh, Yang-Hyo;Park, Yeong-Min
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.482-492
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    • 2007
  • Infection with Shiga-like toxin (SLT)-producing Escherichia coli causes a spectrum of illnesses with high morbidity and mortality. Host mediators play an important role in the pathogenesis of SLT-I toxicity. We here investigated the effect of SLT-I on tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ production, effect of $TNF-{\alpha}$ on glycolipid globotriaosyleramide (Gb3) expression, and relationship between Gb3 level and differential susceptibility of cells to SLT-I. In this study, we observed that detectable levels of $TNF-{\alpha}$ are produced 6 hrs after induction and continued to increase during 48 hrs by SLT-I. It was also found that Vero cells and dendritic cells expressed high levels of Gb3, 83% and 68%, respectively, and that macrophages had a low level of Gb3 (29%) and showed refractory to cytotoxicity against SLT-I. Vero cells and dendritic cells expressing high levels of Gb3 were highly susceptible to SLT-I. furthermore, macrophages showed a resistance to SLT-I cytotoxicity, despite the fact that Gb3 expression was enhanced. These results suggest that the expression of Gb3 is necessary, but not sufficient to confer sensitivity of macrophages to SLT-I and further underpin the important role of SLT-I and its receptor, Gb3, in the pathogenesis of E. coli O157 infection.

Phellinus linteus Extract Regulates Macrophage Polarization in Human THP-1 Cells (상황버섯 추출물의 인간 유래 THP-1 단핵구 세포주의 분극화 조절)

  • Lee, Sang-Yull;Park, Sul-Gi;Yu, Sun-Nyoung;Kim, Ji-Won;Hwang, You-Lim;Kim, Dong-Seob;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.30 no.2
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    • pp.113-121
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    • 2020
  • Macrophages are initiators for regulating a host's defenses to eliminate pathogens and trigger tissue repair. Macrophages are classified into two types: classically (M1) activated macrophages and alternatively (M2) activated macrophages. M1-phenotype macrophages directly or indirectly kill infectious organisms and tumor cells via pro-inflammatory responses, whereas M2-phenotype macrophages remodel wounded tissue through anti-inflammatory responses. In this paper, we investigated how Phellinus linteus hot water extract passed from Diaion HP-20 resin (PLEP) regulates polarization of M1-like or M2-like macrophages in human THP-1 cells. PLEP did not have cytotoxicity at a high concentration of 300 ㎍/ml. We observed morphological alteration of the THP-1 cells, which are stimulated by PLEP, LPS/INF-γ (M1 stimulators) or IL-4/IL13 (M2 stimulators). PLEP exposure induced morphology contiguous with LPS/INF-γ. qPCR was also performed to determine whether PLEP influences M1 or M2 polarization-related genes. M1-phenotype macrophage-specific genes, such as TNF-α, IL-1β, IL-6, IL-8, CXCL10 and CCR7, were enhanced by PLEP in a dose-dependent manner similar to LPS/INF-γ. Conversely, M2-phenotype-specific genes, such as MRC-1, DC-SIGN, CCL17 and CCL22, were suppressed by PLEP. PLEP also significantly up-regulated secretory inflammation cytokines related to M1 polarization of macrophages, including TNFα, IL-1β and IL-6, which was similar to the gene expression. Further, MAPK and NF-κB signaling were increased by treatment with PLEP, resulting in enhancement of cytokine secretion. PLEP might therefore be used as a promising booster of pro-inflammatory responses through M1 polarization of human THP-1 cells.

Seasonal Variations of Environmental Factors and Distribution of Anabaena cylindrica Growth-Inhibiting Bacteria in the Lower Daechung Reservoir (대청호 하류에서 환경요인과 Anabaena cylindrica 생장억제세균의 계절별 분포 변화)

  • Lee, Jung-Ho;Kim, Chul-Ho
    • Korean Journal of Ecology and Environment
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    • v.33 no.2 s.90
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    • pp.128-135
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    • 2000
  • The authors surveyed the seasonal variations of environmental factors, the distributions of heterotrophic bacteria and Anabaena cylindrica growth-inhibiting bacteria at each water layer in Daechung Reservoir to verify the role of bacteria during the extinction of bloom. Average water depth at site 1, 2, and 3 were 25.5 m, 15.0 m and 12.3 m, respectively. Water temperature showed a typical pattern seasonally. The variation of DO was relatively inverse proportional to that of water temperature, although it was irregular during summer time. DO decreased gradually to early May, fluctuated sharply after then, and followed by gradual increasement after middle of September. This variation pattern was notable at surface layer. There was remarkable difference in DO concentraion between surface layer and the other water layers during the period in which DO irregulary varied. The variation range of chlorophyll-a concentraion at surface layer in summer time was broad, and it was relatively high when DO was high. The population size of heterotrophic bacteria was high from Spring to Autumn, an declined after September when the water temperature droped rapidly. Especially this variation pattern was prominent at the surface layer. Bacteria that inhibit the growth of A. cylindrica was almost not detected by June, and its distribution increased in July. Afterward, it showed different variation pattern between each site. The distribution of A. cylindrica growth-inhibiting bacteria was higher at the middle and bottom layer than the surface layer in July and October, when it was larger at all sites for the study period. This result suggests that the antagonistic bacteria exhibit higher activity when host activity drops. These results also suggest that natural water bacteria control the distirbution of cyanobacteria, especially its activity as controller is remarkable when cyanobacterial growth declines.

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Studies on a Nematode, Anguillicola crassa Parasitic in the Air Bladder of the Eel (뱀장어 부레에 기생(寄生)하는 선충(線虫), Anguillicola crassa에 관(關)한 연구(硏究))

  • Kim, Young-Gill;Kim, Eul-Bae;Kim, Jong-Yeon;Chun, Seh-Kyu
    • Journal of fish pathology
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    • v.2 no.1
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    • pp.1-18
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    • 1989
  • In Korea, studies on a Nematode, Anguillicola crassa parasitic in the air bladder of eel are not yet reported. This reason led the author to study the parasitic species, state and life history of the A. crassa parasitized in the air bladder of eel in order to take effective control measures against its damage. The size of fully developed eggs was 80 to $92(86.7){\times}62$ to $71(67.4)\;{\mu}m$, larva was 210 to $240(225){\times}18$ to $23(20.6)\;{\mu}m$. The intermediate host of A. crassa was Thermocyclops hyalinus, it was capable for parasitizing the eel after 4 days of invasion and then the size of larva was 360 to $420(390){\times}28$ to $35(31)\;{\mu}m$. Fifty days after eel had ingested the Thermocyclops hyalinus infected with larva of A. crassa, the larvae matured into adult worms in the air bladder of eel. The size of detected adult worms was 7.3 to $31.0(16.5){\times}0.5$ to 2.2(1.2) mm, 4.9 to $13.3(8.3){\times}0.3$ to 0.9(0.4) mm. Investigating the morphology of the worms, they were identified as A. crassa. Monthly the parasitic rate of the worms in the eel was high in June, September and December, but low in January to March. After the investigation on the significance between non-parasitic fish and parasitic fish, it was not significant, therefore it can be considered that there is no effect of infection in the growth of eel. Any abnormality of eels air bladder tissue was not seen by the infection of A. crassa. At 25.0 to $26.7^{\circ}C$ of water temperature the death time of Thermocyclops hyalinus by masoten treatment was 14 hours in 0.5 ppm, 20 hours in 0.4 ppm, 22 hours in 0.3 ppm, 30 hours in 0.2 ppm and 42 hours in 0.1 ppm.

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Regulation of Tumor Neceosis Factor-${\alpha}$ Receptors and Signal Transduction Pathways

  • Han, Hyung-Mee
    • Toxicological Research
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    • v.8 no.2
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    • pp.343-357
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    • 1992
  • Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

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Fluid Inclusions in Amethyst from the Korea Amethyst Deposit, Uljin, Gyeongbuk (경북 울진 코리아 광상의 자수정에 대한 유체포유물 연구)

  • Lee, Mi-Lyoung;Yang, Kyoung-Hee;Lee, Ju-Youn;Kim, Gyo-Tea
    • Journal of the Mineralogical Society of Korea
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    • v.22 no.3
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    • pp.207-216
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    • 2009
  • Three distinct types of fluid inclusions in amethyst and quartz crystals are associated with metamorphic events in the Korea Amethyst deposit from Uljin-Gun, Gyeongbuk Province. The amethyst displays bimodal grain size distribution in fine-grained, strain-free equigranular quartz with coarse-grained quartz grains with kink bands and undulose extinction. Type I inclusions are liquid-rich and salinity is 0~7 wt% NaCl and the homogenization temperatures ($T_h$) $91{\sim}231^{\circ}C$ with eutectic temperatures ($T_e$) $-52{\sim}-20^{\circ}C$. Type II inclusions are vapor-rich (80~90 vol%). The salinity and $T_h$ ranges 3~6 wt% NaCl and $230{\sim}278^{\circ}C$, respectively with $T_e$ $-56{\sim}-23^{\circ}C$. Type III inclusions contain a daughter mineral other than NaCl. The salinity ranges 32~36 wt% NaCl and $T_h$ $210{\sim}271^{\circ}C$. The textural and fluid inclusion evidences suggest that the host Buncheon granite gneiss and Amethyst pegmatite experienced dynamic recrystallization and the studied fluid inclusions are metamorphic in origin. The metamorphic event possibly occurred at higher temperature than $271{\sim}278^{\circ}C$. The amethysts from Uljin Korea Amethyst can be distinguished from the synthetic amethyst on basis of the distinctive two and three-phases fluid inclusions. Furthermore, it is noticeable that Korea amethyst do not contain NaCl-bearing and $CO_2$-rich fluid inclusions unlike those compared to those from Eonyang and Samcheonpo deposits related to unmetamorphosed granitic rocks.

Construction of a Fluorescently Labeled Infectious R Peptide-Less Moloney MLV Molecular Clone for Analysis of Syncytium (합포체 분석을 위해 R 펩타이드가 결여된 형광 표지 Moloney 마우스레트로바이러스 Molecular Clone 제조)

  • Lee, Yong-Jin;Park, Jin-Woo;Lee, Kyu-Jun;Bae, Eun-Hye;Park, Sung-Han;Lim, Ji-Hyun;Kim, Sae-Ro-Mi;Jung, Yong-Tae
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.246-250
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    • 2009
  • Retroviruses enter host cells by membrane fusion between the viral Env proteins on the virus membrane and a virus receptor on the cellular membrane. The envelope protein of the ecotropic Moloney murine leukemia virus is synthesized as a gp85 precursor and is proteolytically cleaved into an extracellular surface unit (SU) and the transmembrane protein (TM). The cytoplasmic tail (16 amino acid; R peptide) of the TM protein is further cleaved by the viral protease during virion maturation. Unlike the wild type Env protrin bearing the R peptide, R peptide-truncated Envelope induces syncytia in susceptible cells. To understand the mechanism of R peptidetruncated Env in syncytium formation, R peptide-truncated Env expressing full-length molecular clone containing EGFP in PRR (proline rich region) of Env was constructed. This molecular clone induced syncytia in transfected NIH3T3 cells, fluorescence was detected in the cytoplasm and at the plasma membrane, while the nuclei did not stain and appeared black by fluorescence microscopy. Interestingly, virions with truncated envelope produced from transfected NIH3T3 cells induced syncytia in NIH3T3 cells, but fluorescence was not detected in the same infected cells. It is believed that cell-free viruses direct the fusion of neighboring cells without infection. Our data suggests that use of EGFP-tagged envelope for monitoring syncytium is a sensitive and convenient method. We also found that virion incorporated the R peptide-truncated Env is able to induce the formation of syncytia by fusion from without.

Geochemical Characteristics of the Gyeongju LILW Repository II. Rock and Mineral (중.저준위 방사성폐기물 처분부지의 지구화학 특성 II. 암석 및 광물)

  • Kim, Geon-Young;Koh, Yong-Kwon;Choi, Byoung-Young;Shin, Seon-Ho;Kim, Doo-Haeng
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.6 no.4
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    • pp.307-327
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    • 2008
  • Geochemical study on the rocks and minerals of the Gyeongju low and intermediate level waste repository was carried out in order to provide geochemical data for the safety assessment and geochemical modeling. Polarized microscopy, X-ray diffraction method, chemical analysis for the major and trace elements, scanning electron microscopy(SEM), and stable isotope analysis were applied. Fracture zones are locally developed with various degrees of alteration in the study area. The study area is mainly composed of granodiorite and diorite and their relation is gradational in the field. However, they could be easily distinguished by their chemical property. The granodiorite showed higher $SiO_2$ content and lower MgO and $Fe_2O_3$ contents than the diorite. Variation trends of the major elements of the granodiorite and diorite were plotted on the same line according to the increase of $SiO_2$ content suggesting that they were differentiated from the same magma. Spatial distribution of the various elements showed that the diorite region had lower $SiO_2,\;Al_2O_3,\;Na_2O\;and\;K_2O$ contents, and higher CaO, $Fe_2O_3$ contents than the granodiorite region. Especially, because the differences in the CaO and $Na_2O$ distribution were most distinct and their trends were reciprocal, the chemical variation of the plagioclase of the granitic rocks was the main parameter of the chemical variation of the host rocks in the study area. Identified fracture-filling minerals from the drill core were montmorillonite, zeolite minerals, chlorite, illite, calcite and pyrite. Especially pyrite and laumontite, which are known as indicating minerals of hydrothermal alteration, were widely distributed in the study area indicating that the study area was affected by mineralization and/or hydrothermal alteration. Sulfur isotope analysis for the pyrite and oxygen-hydrogen stable isotope analysis for the clay minerals indicated that they were originated from the magma. Therefore, it is considered that the fracture-filling minerals from the study area were affected by the hydrothermal solution as well as the simply water-rock interaction.

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