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ISOLATION OF THE HUMAN CYTOMEGALOVIRUS FROM BODILY FLUIDS Acta biol.Colomb.
CAMACHO ORTEGA,Sigrid Johanna; BOHORQUEZ ÁVILA,Sonia del Pilar; VELANDIA ROMERO,Myriam Lucia; CASTELLANOS PARRA,Jaime Eduardo.
ABSTRACT In vitro studies on the pathogenesis of the human cytomegalovirus (HCMV) are conducted regularly using laboratory adapted strains that lose some characteristics during the adaptation process. Since HCMV is excreted from bodily fluids during infection or reactivation, this work aimed to isolate and culture HCMV from the MRC-5 human cells found in the urine, bronchoalveolar lavage, saliva, and plasma samples of pediatric patients with probable or confirmed infection. The samples were inoculated on cell cultures either for 14 days or until a cytopathic effect (CPE) of 80 % was observed. The cell lysates and supernatants were used to perform successive viral passages. Besides HCMV, the herpes simplex virus was detected from all the saliva samples....
Tipo: Info:eu-repo/semantics/article Palavras-chave: Body fluids; Cell culture; Cytomegalovirus; Isolation.
Ano: 2019 URL: http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S0120-548X2019000300520
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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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In vitro evaluation of Clostridium septicum alpha toxoid Arq. Bras. Med. Vet. Zootec.
Salvarani,F.M.; Lobato,Z.I.P.; Assis,R.A.; Lima,C.G.R.D.; Silva,R.O.S.; Pires,P.S.; Lobato,F.C.F..
Aiming to investigate in vitro alternatives, a test for neutralizing antibody detection using cell culture was developed. This test was more sensitive than previous animal models, allowing for detection of substantially lower alpha toxin and anti-alpha toxin titers. Titers observed during in vivo and in vitro seroneutralization had a correlation of 99.12%, indicating that cell culture is a viable alternative in the evaluation of vaccine potency, screening of vaccinal seeds, and Clostridium septicum alpha toxin titration.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Seroneutralization; Mouse; Guinea pig; Cell culture; Vaccine.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352010000400002
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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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The in vitro comparative cytopathology of a porcine rotavirus and the simian prototype (SA-11) Arq. Bras. Med. Vet. Zootec.
Lonien,S.C.H.; Bolognini,A.M.; Linhares,R.E.C.; Nozawa,C.M..
A citopatologia in vitro de uma cepa de rotavírus porcino adaptado em cultura de células foi comparada à estirpe-protótipo símia (SA-11). O efeito citopático (ECP) produzido pelos vírus foi semelhante embora a estirpe porcina tivesse apresentado algumas alterações diferentes, como o acentuado estreitamento do citoplasma, com grande perda do volume citoplasmático. O vírus porcino apresentou menor número de plaques de ECP porém com diâmetro maior em relação ao vírus símio, demonstrando maior capacidade de disseminação célula-célula, quase oito vezes mais, a julgar pelo diâmetro dos plaques de ECP. Os elementos do citoesqueleto das células infectadas revelaram uma reorganização semelhante para ambas as estirpes, não sendo possível observar nenhuma diferença,...
Tipo: Info:eu-repo/semantics/report Palavras-chave: Rotavirus; Cytopathology; Cell culture; Plaque; Cytoskeleton.
Ano: 2001 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352001000400002
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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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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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The effect of concanavalin A on the replication of rotavirus (SA-11) in cell culture BABT
Hasenack,Beatriz S.; Botelho,Maria Valéria J.; Lauretti,Flávio; Melo,Fernando L. de; Orlandi,Janaina M.; Linhares,Rosa Elisa C.; Ueda,Marli; Nozawa,Carlos M..
Rotavirus (RV) strain SA-11 was studied with respect to its infectivity in MA-104 cell cultures and the effect of concanavalin A (ConA). Receptors for ConA at the surface of MA-104 cell were determined by fluorescence assay and specifically inhibited by D-mannose. The kinetics of virus growth was carried out by plaque assay. Electron microscopy and polyacrylamide gel electrophoresis were used for monitoring the experiments. It was concluded that RV replication was not affected consistently by ConA, however it interfered with the development of cytopathic effect (CPE) without altering virus yields.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Rotavirus; Concanavalin A; Cell culture.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132002000200003
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Isolation in HRT-18 cells and molecular analysis of a BCoV strain from diarrheic feces of naturally infected calves BABT
Stipp,Danilo Tancler; Barry,Aline Fernandes; Alfieri,Alice Fernandes; Bodnar,Lívia; Alfieri,Amauri Alcindo.
Bovine coronavirus (BCoV) may cause acute diarrhea in newborn calves, leading to significant economic losses for cattle farmers. There are several diagnostic techniques used to detect BCoV in calf fecal samples, but virus isolation still has advantages for antigenic and genomic characterization. This study describes the isolation in HRT-18 cells and molecular characterization of Brazilian BCoV wild-type strains. Three fecal samples from diarrheic 30 day-old calves were inoculated in HRT-18 cell monolayers, which were then evaluated for HA titers and tested using semi-nested PCR followed by RFLP and sequencing. Two samples were successfully isolated and presented HA titers of 16 and 32 units per 25 mL. The results were confirmed using semi-nested PCR and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cattle; Diarrhea; Bovine coronavirus; Cell culture; Molecular analyses.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000700007
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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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Primary culture of hepatic cells from Metynnis roosevelti (Pisces, Teleostei, Characidae) Braz. J. Vet. Res. Anim. Sci.
SALVO,Ligia Maria; MALUCELLI,Maria Ivette Carboni; RICHARTZ,Rosária Regina Tesoni B.; BACILA,Metry.
In order to achieve monolayers and adhesion in cell cultures of hepatocytes from the subtropical fish Metynnis roosevelti (Pisces, Teleostei, Characidae) three different methodologies were used: enzyme cell dissociation by means of a system containing trypsin and versene 0.25%; by means of bovine fetal serum and by the use of blood serum obtained from adult specimens of M. roosevelti. The cell medium used has been a modified F10-199 by the addition of 2 mM L-glutamine, 10 µg/mL insulin, 50 µg/mL fibronectin; and antibiotics. The best results in terms of cell dissociation as well as in terms of adhesion and confluence of the cells monolayer has been achieved by means of the enzymatic cell dissociation.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Liver cells; Cell culture; Cells; Pisces; Metynnis roosevelti.
Ano: 2000 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-95962000000500004
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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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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
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Supraorganized collagen enhances Schwann cell reactivity and organization in vitro BJMBR
Maturana,L.G.; Zanon,R.G.; Pierucci,A.; Vidal,B.C.; Oliveira,A.L.R..
We investigated the reactivity and expression of basal lamina collagen by Schwann cells (SCs) cultivated on a supraorganized bovine-derived collagen substrate. SC cultures were obtained from sciatic nerves of neonatal Sprague-Dawley rats and seeded on 24-well culture plates containing collagen substrate. The homogeneity of the cultures was evaluated with an SC marker antibody (anti-S-100). After 1 week, the cultures were fixed and processed for immunocytochemistry by using antibodies against type IV collagen, S-100 and p75NTR (pan neurotrophin receptor) and for scanning electron microscopy (SEM). Positive labeling with antibodies to the cited molecules was observed, indicating that the collagen substrate stimulates SC alignment and adhesion (collagen IV...
Tipo: Info:eu-repo/semantics/article Palavras-chave: S-100; P75NTR; Type IV collagen; Collagen; Cell culture; Supraorganized collagen.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2011000700011
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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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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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Use of blends of bioabsorbable poly(L-lactic acid)/poly(hydroxybutyrate- co-hydroxyvalerate) as surfaces for Vero cell culture BJMBR
Santos Jr.,A.R.; Ferreira,B.M.P.; Duek,E.A.R.; Dolder,H.; Wada,M.L.F..
Vero cells, a cell line established from the kidney of the African green monkey (Cercopithecus aethiops), were cultured in F-10 Ham medium supplemented with 10% fetal calf serum at 37°C on membranes of poly(L-lactic acid) (PLLA), poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and their blends in different proportions (100/0, 60/40, 50/50, 40/60, and 0/100). The present study evaluated morphology of cells grown on different polymeric substrates after 24 h of culture by scanning electron microscopy. Cell adhesion was also analyzed after 2 h of inoculation. For cell growth evaluation, the cells were maintained in culture for 48, 120, 240, and 360 h. For cytochemical study, the cells were cultured for 120 or 240 h, fixed, processed for histological analysis,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Poly(L-lactic acid); Poly(hydroxybutyrate- co-hydroxyvalerate); Blends; Biomaterials; Cell culture; Vero cells.
Ano: 2005 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2005001100009
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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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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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