NÉLIO ALESSANDRO DE JESUS OLIVEIRA

NÉLIO ALESSANDRO DE JESUS OLIVEIRA

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Phenotype changes of oral epithelial stem cells after in vitro culture
2020 - DALTOE, FELIPE P.; OLIVEIRA, NÉLIO A.J. de; PERONI, CIBELE N.; SHARPE, PAUL T.; MANTESSO, ANDREA
The aim of our study was to isolate populations of keratinocyte stem cells based on the expression of cell surface markers and to investigate whether the culture could affect their phenotype. keratinocytes from human oral mucosa were sorted based on the expression of the epithelial stem cell markers p75NTR and CD71. We also examined the co-expression of other epithelial stem markers such as integrins β1 and α6 and their stem cell-like proprieties in in vitro assays. Three passages after being sorted by MACS, more than 93% of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of the p75NTR-ve gained it. Within the small population of the p75NTR+ve cells, 88% co-expressed other epithelial stem cell markers such as integrins β1 and α6, while only 28% of p75NTR-ve cells co-expressed these markers. These results were confirmed by sorting cells by FACS. Additionally, when double staining was used for sorting cells, 99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells expressed both integrins, but just one week after culture, only 1.74% of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32% still expressed CD71. Similar results were obtained when co-culturing p75NTR+ve and p75NTR-ve populations before analysis. Our results suggest that phenotype changes may be part of an intrinsic cellular mechanism to conserve levels of protein expression as they may found in the human body. In addition, in vitro culture may not offer ideal conditions for epithelial stem cell maintenance due to phenotype changes under standard culture conditions.
Molecular cloning and characterization of pirarucu (Arapaima gigas) follicle-stimulating hormone and luteinizing hormone β-subunit cDNAs
2017 - SEVILHANO, THAIS; CARVALHO, ROBERTO F. de; OLIVEIRA, NELIO A. de J.; OLIVEIRA, JOAO E.; MALTAROLLO, VINICIUS G.; TROSSINI, GUSTAVO; GARCEZ, RIVIANE; BARTOLINI, PAOLO
The common gonadotrophic hormone α-subunit (GTHα) has been previously isolated by our research group from A. gigas pituitaries; in the present work the cDNA sequences encoding FSHβ and LHβ subunits have also been isolated from the same species of fish. The FSH β-subunit consists of 126 amino acids with a putative 18 amino acid signal peptide and a 108 amino acid mature peptide, while the LH β-subunit consists of 141 amino acids with a putative 24 amino acid amino acid signal peptide and a 117 amino acid mature peptide. The highest identity, based on the amino acid sequences, was found with the order of Anguilliformes (61%) for FSHβ and of Cypriniformes (76%) for LHβ, followed by Siluriformes, 53% for FSHβ and 75% for LHβ. Interestingly, the identity with the corresponding human amino acid sequences was still remarkable: 45.1% for FSHβ and 51.4% for LHβ. Three dimensional models of ag-FSH and ag-LH, generated by using the crystal structures of h-FSH and h-LH as the respective templates and carried out via comparative modeling and molecular dynamics simulations, suggested the presence of the so-called ªseat-beltº, favored by a disulfide bond formed between the 3rd and 12th cysteine in both β-subunits. The sequences found will be used for the biotechnological synthesis of A. gigas gonadotrophic hormones (ag-FSH and ag-LH). In a first approach, to ascertain that the cloned transcripts allow the expression of the heterodimeric hormones, ag-FSH has been synthesized in human embryonic kidney 293 (HEK293) cells, preliminarily purified and characterized.
Partial correction of the dwart phenotype by non-viral transfer of the growth hormone gene in mice: treatment age is critical
2016 - HIGUTI, ELIZA; CECCHI, CLAUDIA R.; OLIVEIRA, NELIO A.J.; LIMA, ELIANA R.; VIEIRA, DANIEL P.; AAGAARD, LARS; JENSEN, THOMAS G.; JORGE, ALEXANDER A.L.; BARTOLINI, PAOLO; PERONI, CIBELE N.
The influence of Escherichia coli cultivation temperature on interferon alpha 2a expression (IFN-'alfa'2a)
2014 - ARTHUSO, FERNANDA dos S.; SUZUKI, MIRIAM F.; OLIVEIRA, NELIO A. de J.; OLIVEIRA, JOAO E. de; BARTOLINI, PAOLO; SOARES, CARLOS R.J.
Different ex vivo and direct in vivo DNA administration strategies for growth hormone gene therapy in dwarf animals
2011 - PERONI, CIBELE N.; OLIVEIRA, NELIO A. de J.; CECCHI, CLAUDIA R.; HIGUTI, ELIZA; BARTOLINI, PAOLO
In vitro, ex vivo and in vivo recombinant growth hormone synthesis, for gene therapy applications
2011 - PERONI, CIBELE N.; OLIVEIRA, NELIO A. de J.; CECCHI, CLAUDIA R.; HIGUTI, ELIZA; SILVA, JULIANA T. da; JORGE, ALEXANDER A. de L.; JENSEN, THOMAS G.; BARTOLINI, PAOLO
Growth parameters after intramuscular hGH plasmid administration compared to recombinant hGH injections in LIT/SCID mice
2011 - HIGUTI, E.; CECCHI, C.R.; OLIVEIRA, N.A.J.; SILVA, J.T.; JORGE, A.A.L.; BARTOLINI, P.; PERONI, C.N.
Assessment of the in vivo permanence of mGH-secreting human keratinocytes in grafted organotypic cultures
2009 - CECCHI, C.R.; OLIVEIRA, N.A.J.; HIGUTI, E.; NONOGAKI, S.; BOCCARDO, E.; BARTOLINI, P.; PERONI, C.N.
Electrotransfer of naked hGH cDNA muscle of lit/scid mouse is more efficient than genomic DNA
2009 - OLIVEIRA, N.A.J.; CECCHI, C.R.; HIGUTI, E.; MORISCOT, A.S.; BARTOLINI, P.; PERONI, C.N.
Sustained hGH expression after electrotransfer of naked DNA into DWARF 'little!' mouse skeletal muscle
2009 - OLIVEIRA, N.A.J.; CECCHI, C.R.; HIGUTI, E.; BARTOLINI, P.; PERONI, C.N.