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A simple and effective pressure culture system modified from a transwell cell culture system Biol. Res.
Sun,Xuerong; Liu,Xinguang; Zhang,Yuehong; Kuang,Xielan; Lv,Bo; Ge,Jian.
Mechanical pressure plays an important role in many physiological and pathological processes. Mimicking the mechanical pressure present in vitro is necessary for related research, but usually requires expensive and complicated equipment. In this study we created a simple pressure culture system based on the transwell culture system. By cutting off the top rim of the transwell insert, the cells were compressed between the insert membrane and the well floor. The new pressure culture system was proven effective in that it induced cell morphological change, integrin β1 upregulation, actin polymerization and growth change in rat retinal ganglion cells, human nasopharyngeal carcinoma cells and mice embryonic fibroblasts. Though the pressure value is...
Tipo: Journal article Palavras-chave: Mechanical pressure; Cell culture; Transwell culture system; Cytoskeleton.
Ano: 2013 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602013000100007
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Adenosine as a signaling molecule in the retina: biochemical and developmental aspects Anais da ABC (AABC)
PAES-DE-CARVALHO,ROBERTO.
The nucleoside adenosine plays an important role as a neurotransmitter or neuromodulator in the central nervous system, including the retina. In the present paper we review compelling evidence showing that adenosine is a signaling molecule in the developing retina. In the chick retina, adenosine transporters are present since early stages of development before the appearance of adenosine A1 receptors modulating dopamine-dependent adenylate cyclase activity or A2 receptors that directly activate the enzyme. Experiments using retinal cell cultures revealed that adenosine is taken up by specific cell populations that when stimulated by depolarization or neurotransmitters such as dopamine or glutamate, release the nucleoside through calcium-dependent...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Nucleoside; Receptors and transporters; Development; Cell culture; Uptake and release.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652002000300007
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Amniotic cell culture during different ages of gestation for karyotype analysis in bovine Braz. J. Vet. Res. Anim. Sci.
EIRAS,Paola Ribeiro Seabra; BARRETO FILHO,João Bosco; GOLGHER,Romain Rolland; SANTOS,Sandra Regina Quintino dos.
Bovine karyotyping has become an important diagnostic tool in animal breeding. In the prenatal period it can diagnose several chromosomal abnormalities such as Robertsonian translocations, testicle feminization syndrome, gonadal dysgenesis and Klinefelter’s syndrome. An important cell source for karyotype analysis is the amniotic fluid. It has been extensively used in humans but in bovine, however, this is not the case despite its diagnostic value. Since a small percentage of cells is viable, cells and their growth conditions as well as the handling of the material should be optimal to insure a successful analysis. For this, we have compared the growth efficiency for bovine amniocytes in two media, employing cells from 10 to 14 weeks of gestation....
Tipo: Info:eu-repo/semantics/article Palavras-chave: Karyotypes; Cell culture; Amnion; Cattle.
Ano: 2000 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-95962000000400005
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Biological activity of Serratia marcescens cytotoxin BJMBR
Carbonell,G.V.; Amorim,C.R.N.; Furumura,M.T.; Darini,A.L.C.; Fonseca,B.A.L.; Yano,T..
Serratia marcescens cytotoxin was purified to homogeneity by ion-exchange chromatography on a DEAE Sepharose Fast Flow column, followed by gel filtration chromatography on a Sephadex G100 column. The molecular mass of the cytotoxin was estimated to be about 50 kDa. Some biological properties of the cytotoxin were analyzed and compared with well-characterized toxins, such as VT1, VT2 and CNF from Escherichia coli and hemolysin produced by S. marcescens. The sensitivity of the cell lines CHO, HeLa, HEp-2, Vero, BHK-21, MA 104 and J774 to the cytotoxin was determined by the cell viability assay using neutral red. CHO and HEp-2 were highly sensitive, with massive cellular death after 1 h of treatment, followed by BHK-21, HeLa, Vero and J774 cells, while MA 104...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cell culture; Biological activity; Serratia marcescens; Cytotoxin; Virulence factors.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003000300010
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Biosynthesis and metabolism of steroid hormones by human adrenal carcinomas BJMBR
Brown,J.W.; Fishman,L.M..
Over a 15-year period, our university-based laboratory obtained 125 adrenal tumors, of which 15 (12%) were adrenal cortical carcinomas. Of these, 6 (40% of the carcinomas) occurred in patients with clear clinical manifestations of steroid hormone excess. Adrenal cortical carcinoma cells derived from the surgically resected tumors in 4 of these patients were isolated and established in primary culture. Radiotracer steroid interconversion studies were carried out with these cultures and also on mitochondria isolated from homogenized tissues. Large tumors had the lowest steroidogenic activities per weight, whereas small tumors had more moderately depressed enzyme activities relative to cells from normal glands. In incubations with pregnenolone as substrate, 1...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Adrenal carcinoma; Cell culture; Steroids; Metyrapone; Dibutyryl cAMP; ACTH; Krebs cycle intermediates.
Ano: 2000 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2000001000014
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Bivale mollusc cell culture ArchiMer
Mialhe, Eric; Boulo, Viviane; Grizel, Henri.
The historical background of bivalve cell culture and its potential uses in molluscan pathology are reviewed. Then, primary cultures for the study of some host-pathogen interactions at the cellular level are considered. Next, alternative methods to classical cell cultures are examined that may lead to the production of bivalve celllines.
Tipo: Text Palavras-chave: Cell culture; Pathology; Molluscs.
Ano: 1988 URL: http://archimer.ifremer.fr/doc/1988/publication-3122.pdf
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Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3 BJMBR
Liu,Z.; Zhang,P.; Zhou,Y.; Qin,H.; Shen,T..
Epithelium, a highly dynamic system, plays a key role in the homeostasis of the intestine. However, thus far a human intestinal epithelial cell line has not been established in many countries. Fetal tissue was selected to generate viable cell cultures for its sterile condition, effective generation, and differentiated character. The purpose of the present study was to culture human intestinal epithelial cells by a relatively simple method. Thermolysin was added to improve the yield of epithelial cells, while endothelin-3 was added to stimulate their growth. By adding endothelin-3, the achievement ratio (viable cell cultures/total cultures) was enhanced to 60% of a total of 10 cultures (initiated from 8 distinct fetal small intestines), allowing the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Human intestinal epithelial cell; Cell culture; Thermolysin; Endothelin-3; Cytokeratin; Brush border enzyme.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000500006
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Cytoskeleton, endoplasmic reticulum and nucleus alterations in CHO-K1 cell line after Crotalus durissus terrificus (South American rattlesnake) venom treatment J. Venom. Anim. Toxins incl. Trop. Dis.
Tamieti,B. P.; Damatta,R. A.; Cogo,J. C.; Da Silva,N. S.; Mittmann,J.; Pacheco-Soares,C..
Snake venoms are toxic to a variety of cell types. However, the intracellular damages and the cell death fate induced by venom are unclear. In the present work, the action of the South American rattlesnake Crotalus durissus terrificus venom on CHO-K1 cell line was analyzed. The cells CHO-K1 were incubated with C. d. terrificus venom (10, 50 and 100g/ml) for 1 and 24 hours, and structural alterations of actin filaments, endoplasmic reticulum and nucleus were assessed using specific fluorescent probes and agarose gel electrophoresis for DNA fragmentation. Significant structural changes were observed in all analyzed structures. DNA fragmentation was detected suggesting that, at the concentrations used, the venom induced apoptosis.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cell culture; Actin filaments; Nucleus; Endoplasmic reticulum; Crotalus; Rattlesnake venom; Apoptosis.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992007000100004
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Detection of multiple mycoplasma infection in cell cultures by PCR BJMBR
Timenetsky,J.; Santos,L.M.; Buzinhani,M.; Mettifogo,E..
A total of 301 cell cultures from 15 laboratories were monitored for mycoplasma (Mollicutes) using PCR and culture methodology. The infection was detected in the cell culture collection of 12 laboratories. PCR for Mollicutes detected these bacteria in 93 (30.9%) samples. Although the infection was confirmed by culture for 69 (22.9%) samples, PCR with generic primers did not detect the infection in five (5.4%). Mycoplasma species were identified with specific primers in 91 (30.2%) of the 98 samples (32.6%) considered to be infected. Mycoplasma hyorhinis was detected in 63.3% of the infected samples, M. arginini in 59.2%, Acholeplasma laidlawii in 20.4%, M. fermentans in 14.3%, M. orale in 11.2%, and M. salivarium in 8.2%. Sixty (61.2%) samples were...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mycoplasma; Cell culture; Multiple infection; Mycoplasma detection.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000700009
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Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR BJM
Vinatea,Cecília E.B.; Sincero,Thaís C.M.; Simões,Claúdia M.O.; Barardi,Célia R.M..
Shellfish are readily contaminated with viruses present in water containing sewage due to the concentrating effect of filter feeding. Enteroviruses are generally used as a model for the detection of viruses from shellfish due to their public health significance. In the present work, oysters were placed in glass aquaria containing seawater plus unicellular algae. Two experiments were performed: 1) oysters bioaccumulating four different poliovirus type 2 concentrations: 5 x 10(4), 2.5 x 10(4), 5 x 10³ and 5 x 10² PFU/mL during 20h; 2) oyster tissues directly inoculated with 6.0 x 10(5) and 1.0 x 10(5) PFU/mL. After viruses seeding, tissue samples were processed by an adsorption-elution-precipitation method. Positive controls were performed by seeding 6.0 x...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Oysters; Poliovirus; Bioaccumulation; Cell culture; RT-PCR.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822006000100012
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Development of mussel-specific expression vectors suitable for transgenic organisms: implication on the establishment of mollusc continuous cell lines ArchiMer
Delsert, Claude; Cancela Da Fonseca, Leonor.
The main objective of this project was to develop the tools for the in-depth understanding of mollusc biology and pathology. Such a goal has been reached in other biological systems through the development of molecular biology and cell culture. Indeed, development of a mollusc cell culture system is an essential step to approach many of the problems related to mollusc pathology, in particular to use molecular biology tools to study viral infection of these organisms. No continous cell line is available from bivalves and while cell cultures have been obtained from these organisms they are difficult to develop and labor intensive, survival cultures being maintained only for a few weeks. Our goal has been to study in detail many of the parameters involved in...
Tipo: Text Palavras-chave: Pathology; Mussel-specific expression vectors; Cell culture; Mytilus galloprovincialis; Gene; Genetic marker.
Ano: 1998 URL: http://archimer.ifremer.fr/doc/00044/15540/12917.pdf
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Diversity of enterovirus sequences detected in oysters by RT-heminested PCR ArchiMer
Dubois, Eric; Merle, Ghislaine; Roquier, Catherine; Trompette, Aurélien; Le Guyader, Soizick; Cruciere, Catherine; Chomel, Jean-jacques.
Oysters harvested in western France, from five sites associated with outbreaks of food-borne norovirus gastroenteritis between February 2000 and March 2001, were assayed for enterovirus RNA by reverse transcriptase-heminested polymerase chain reaction (RT-heminested PCR). Forty percent (21/52) of shellfish samples (pool of seven oysters) were contaminated by enteroviruses. Infectious coxsackieviruses serotype A21 were isolated from three of these positive samples. Amplicons corresponding to 65 base sequences in the 5' untranslated region of the enteroviral genome were analyzed by direct sequencing. Interpretable results were obtained from 18 amplicons, but mixtures of sequences confused the results from 3 samples. Sequences isolated from samples from the...
Tipo: Text Palavras-chave: Sequence analyze; RT PCR; Cell culture; Shellfish; Enterovirus.
Ano: 2004 URL: http://archimer.ifremer.fr/doc/2004/publication-763.pdf
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Effect of actinomycin D on simian rotavirus (SA11) replication in cell culture BJMBR
Stefanelli,C.C.; Castilho,J.G.; Botelho,M.V.J.; Linhares,R.E.C.; Nozawa,C.M..
Rotaviruses are the major cause of viral diarrhea in humans and animals. Actinomycin D (Act D) is an antibiotic that intercalates DNA and therefore inhibits DNA-dependent transcription. The current study was carried out to assess the influence of Act D on the replication of simian rotavirus (SA11) in cell culture. Virus-infected MA-104 cell cultures were studied in the presence of Act D at concentrations of 1.25 and 2.5 µg/ml. Treatment of rotavirus-infected cells with 2.5 µg/ml Act D 48 h post-infection reduced the cytoplasmic metachromasia after staining with acridine orange by 25%. Viral RNA labeled with ³H-uridine in the presence of the drug was separated by polyacrylamide gel electrophoresis. Viral RNA replication was not affected by Act D, but...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Rotavirus; Actinomycin D; Replication; Cell culture.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2002000400006
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Effect of amikacin, cephalothin, clindamycin and vancomycin on in vitro fibroblast growth Genet. Mol. Biol.
Souza,Fernanda Timm Seabra; Mello,Alexandre Silva de; Michelin,Kristiane; Coelho,Janice Carneiro.
The effect of four antibiotics (amikacin, clindamycin, cephalothin and vancomycin) was investigated considering that bacterial infection in fibroblasts cultures is a very frequent event. The investigation included the effect of the antibiotics on fibroblast growth and on the activity of the enzyme glucocerebrosidase. The antibiotics were added to the fibroblast cultures and cell growth was evaluated by counting the number of cells and their viability. After cell harvesting, the enzyme activity and content of protein were measured. The results allowed us to conclude that none of the antibiotics affected the cellular number nor the cellular viability. The content of protein decreased when cephalothin and clindamycin were added to the cultures, and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Amikacin; Cephalothin; Clindamycin; Vancomycin; Fibroblasts; Cell culture.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572004000300023
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Evaluation of a multiplex PCR method to detect enteroaggregative Escherichia coli Biocell
Rüttler,M.E.; Yanzón,C.S.; Cuitiño,M.J.; Renna,N.F.; Pizarro,M.A.; Ortiz,A.M..
Enteroaggregative Escherichia coli (EAEC) has been implicated in sporadic diarrhea in children and adults and has been identified as the cause of several outbreaks worldwide. The HEp-2 test remains the gold standard for identification of this pathotype. A 60-65 MDa plasmid encodes the aggregative adherence fimbriae (AAF/I and AAF/II), a transcriptional activator ( aggR gene), the enteroaggregative heat-stable enterotoxin EAST1 ( astA gene) and a cytotoxin (Pet). The standard assay for EAEC is performed only in research laboratories, because it is expensive, labor intensive and time-consuming. The Polymerase Chain Reaction (PCR) offers the possibility of rapid diagnosis. In the current study, a multiplex PCR assay which checks aggR and astA genes was...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Diarrhea; Escherichia coli; Cell culture; PCR (Polymerase Chain Reaction).
Ano: 2006 URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S0327-95452006000200006
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Fingerprinting of cell lines by directed amplification of minisatellite-region DNA (DAMD) BJMBR
Silva,L.M.; Montes de Oca,H.; Diniz,C.R.; Fortes-Dias,C.L..
The development of in vitro propagation of cells has been an extraordinary technical advance for several biological studies. The correct identification of the cell line used, however, is crucial, as a mistaken identity or the presence of another contaminating cell may lead to invalid and/or erroneous conclusions. We report here the application of a DNA fingerprinting procedure (directed amplification of minisatellite-region DNA), developed by Heath et al. [Nucleic Acids Research (1993) 21: 5782-5785], to the characterization of cell lines. Genomic DNA of cells in culture was extracted and amplified by PCR in the presence of VNTR core sequences, and the amplicons were separated by agarose gel electrophoresis. After image capture with a digital camera, the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: DNA fingerprinting; DAMD; Cell line; Cell culture.
Ano: 2001 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001001100005
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Goat umbilical cord cells are permissive to small ruminant lentivirus infection in vitro BJM
Martins,Gabrielle R.; Marinho,Rebeca C.; Bezerra Junior,Rosivaldo Q.; Alves,Antoniel de O.; Câmara,Lilia M.C.; Albuquerque-Pinto,Luiz C.; Teixeira,Maria F. da S..
Abstract Small ruminant lentiviruses isolated from peripheral blood leukocytes and target organs can be propagated in vitro in fibroblasts derived from goat synovial membrane cells. These cells are obtained from tissues collected from embryos or fetuses and are necessary for the establishment of the fibroblast primary culture. A new alternative type of host cells, derived from goat umbilical cord, was isolated and characterized phenotypically with its main purpose being to obtain cell monolayers that could be used for the diagnosis and isolation of small ruminant lentiviruses in cell culture. To accomplish this goal, cells were isolated from umbilical cords; characterized phenotypically by flow cytometry analysis; differentiate into osteogenic,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: CAEV; MVV; Cell culture; MSC; Flow cytometry.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000100125
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Immunophenotyping, plasticity tests and nanotagging of stem cells derived from adipose tissue of wild rodent agouti (Dasyprocta prymnolopha) Arq. Bras. Med. Vet. Zootec.
Rocha,A.R.; Leite,Y.K.C.; Silva,A.S.; Conde Júnior,A.M.; Costa,C.R.M.; Silva,G.C.; Bezerra,D.O.; Cavalcante,M.M.A.S.; Feitosa,M.L.T.; Argôlo Neto,N.M.; Serakides,R.; Carvalho,M.A.M..
ABSTRACT There is a growing interest in the study of unspecialized mesenchymal stem cells, for there are still some discussions about their in vitro behavior. Regenerative medicine is a science undergoing improvement which develops treatments as cell therapy using somatic stem cells. In several studies, adipose tissue is presented as a source of multipotent adult cells that has several advantages over other tissue sources. This study aimed to characterize and evaluate the tagging of mesenchymal stem cells from the agoutis adipose tissue (Dasyprocta prymonolopha), with fluorescent intracytoplasmic nanocrystals. Fibroblast cells were observed, plastic adherent, with extended self-renewal, ability to form colonies, multipotency by differentiation into three...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cell culture; Mesenchymal; Stem cells; Adipose tissue; Cell differentiation; Fluorescent nanocrystals.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352019000501571
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In vitro cultivation of canine multipotent mesenchymal stromal cells on collagen membranes treated with hyaluronic acid for cell therapy and tissue regeneration BJMBR
Wodewotzky,T.I.; Lima-Neto,J.F.; Pereira-Júnior,O.C.M.; Sudano,M.J.; Lima,S.A.F.; Bersano,P.R.O.; Yoshioka,S.A.; Landim-Alvarenga,F.C..
Support structures for dermal regeneration are composed of biodegradable and bioresorbable polymers, animal skin or tendons, or are bacteria products. The use of such materials is controversial due to their low efficiency. An important area within tissue engineering is the application of multipotent mesenchymal stromal cells (MSCs) to reparative surgery. The combined use of biodegradable membranes with stem cell therapy may lead to promising results for patients undergoing unsuccessful conventional treatments. Thus, the aim of this study was to test the efficacy of using membranes composed of anionic collagen with or without the addition of hyaluronic acid (HA) as a substrate for adhesion and in vitro differentiation of bone marrow-derived canine MSCs. The...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Collagen scaffolds; Biomaterials; Hyaluronic acid; Mesenchymal stromal cells; Tissue engineering; Cell culture.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2012001200008
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In vitro culture and characterization of alveolar bone osteoblasts isolated from type 2 diabetics BJMBR
Sun,Dao-Cai; Li,De-Hua; Ji,Hui-Cang; Rao,Guo-Zhou; Liang,Li-Hua; Ma,Ai-Jie; Xie,Chao; Zou,Gui-Ke; Song,Ying-Liang.
In order to understand the mechanisms of poor osseointegration following dental implants in type 2 diabetics, it is important to study the biological properties of alveolar bone osteoblasts isolated from these patients. We collected alveolar bone chips under aseptic conditions and cultured them in vitro using the tissue explants adherent method. The biological properties of these cells were characterized using the following methods: alkaline phosphatase (ALP) chemical staining for cell viability, Alizarin red staining for osteogenic characteristics, MTT test for cell proliferation, enzyme dynamics for ALP contents, radio-immunoassay for bone gla protein (BGP) concentration, and ELISA for the concentration of type I collagen (COL-I) in the supernatant....
Tipo: Info:eu-repo/semantics/article Palavras-chave: Alveolar bone; Cell culture; Implant; Osteoblast; Type 2 diabetics.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2012000600005
Registros recuperados: 35
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