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1.
Int J Parasitol ; 45(8): 527-35, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25907768

RESUMO

The invertebrate cell line, Bge, from embryos of the snail Biomphalaria glabrata, remains to date the only established cell line from any species of the Phylum Mollusca. Since its establishment in 1976 by Eder Hansen, few studies have focused on profiling its cytometrics, growth characteristics or sensitivity to xenobiotics. Bge cells are reputed to be challenging to propagate and maintain. Therefore, even though this cell line is a noteworthy resource, it has not been studied widely. With growing interest in functional genomics, including genetic transformation, to elucidate molecular aspects of the snail intermediate hosts responsible for transmission of schistosomiasis, and aiming to enhance the convenience of maintenance of this molluscan cell line, we deployed the xCELLigene real time approach to study Bge cells. Doubling times for three isolates of Bge, termed CB, SL and UK, were longer than for mammalian cell lines - longer than 40 h in complete Bge medium supplemented with 7% fetal bovine serum at 25°C, ranging from ∼42 h to ∼157 h when 40,000 cells were seeded. To assess the potential of the cells for genetic transformation, antibiotic selection was explored. Bge cells were sensitive to the aminonucleoside antibiotic puromycin (from Streptomyces alboniger) from 5 µg/ml to 200 ng/ml, displaying a half maximal inhibitory concentration (IC50) of ∼1.91 µg/ml. Sensitivity to puromycin, and a relatively quick kill time (<48 h in 5 µg/ml) facilitated use of this antibiotic, together with the cognate resistance gene (puromycin N-acetyl-transferase) for selection of Bge cells transformed with the PAC gene (puroR). Bge cells transfected with a plasmid encoding puroR were partially rescued when cultured in the presence of 5 µg/ml of puromycin. These findings pave the way for the development of functional genomic tools applied to the host-parasite interaction during schistosomiasis and neglected tropical trematodiases at large.


Assuntos
Antibacterianos/farmacologia , Biomphalaria/genética , Schistosoma mansoni/fisiologia , Animais , Biomphalaria/efeitos dos fármacos , Biomphalaria/embriologia , Biomphalaria/parasitologia , Linhagem Celular , Interações Hospedeiro-Parasita , Puromicina/farmacologia , Xenobióticos/farmacologia
2.
J Gen Microbiol ; 128(2): 423-5, 1982 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-7042905

RESUMO

Mycobacterium leprae isolated from armadillo tissue incorporated radioactivity from D-(14C)glucose and (14C)protein hydrolysate. In the presence of glucose, the rate of incorporation of (14C)protein hydrolysate was increased. Uptake of glucose was inhibited by 2-deoxy-D-glucose and sodium azide; that of the amino acids was inhibited by puromycin and chloramphenicol and, weakly, by cycloheximide.


Assuntos
Aminoácidos/metabolismo , Glucose/metabolismo , Mycobacterium leprae/metabolismo , Azidas/farmacologia , Proteínas de Bactérias/biossíntese , Transporte Biológico , Cloranfenicol/farmacologia , Cicloeximida/farmacologia , Cinética , Puromicina/farmacologia , Azida Sódica
3.
s.l; s.n; feb. 1982. 3 p. graf.
Não convencional em Inglês | SES-SP, HANSEN, SESSP-ILSLACERVO, SES-SP | ID: biblio-1240616

RESUMO

Mycobacterium leprae isolated from armadillo tissue incorporated radioactivity from D-(14C)glucose and (14C)protein hydrolysate. In the presence of glucose, the rate of incorporation of (14C)protein hydrolysate was increased. Uptake of glucose was inhibited by 2-deoxy-D-glucose and sodium azide; that of the amino acids was inhibited by puromycin and chloramphenicol and, weakly, by cycloheximide.


Assuntos
Aminoácidos/metabolismo , Azida Sódica , Cinética , Cloranfenicol/farmacologia , Mycobacterium leprae/metabolismo , Proteínas de Bactérias/biossíntese , Puromicina/farmacologia , Transporte Biológico
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