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A Goethals

Publications and source records attributed to A Goethals.

8 recordsLinked to original sources

SSCP analysis of the P2 purine transporter TcoAT1 gene of Trypanosoma congolense leads to a simple PCR-RFLP test allowing the rapid identification of diminazene resistant stocks.

Analyses were made on a Trypanosoma congolense contig coding a putative P2-like nucleoside transporter (the contig was named in this study TcoAT1). The sequence includes a start and stop codon and presents a high similarity with the gene TbAT1 of T. brucei (Smallest Sum Probability 2.8e-136). To investigate a possible link between point mutations and diminazene aceturate (DA) resistance in mice, the TcoAT1 putative genes of 26 T. congolense strains, characterised for DA sensitivity in the single dose mouse test, were screened by means of the Single Strand Conformation Polymorphism technique (SSCP). Results showed that the SSCP profiles of 23 out of 26 (88.5%) T. congolense strains were confirmed by the sensitivity test in mice with the commonly accepted criterion for sensitivity to diminazene being a CD80 of 20mg/kg in the mouse test. The remaining T. congolense strains showed a resistant SSCP profile and relapsed in mice after treatment at doses lower than 20mg/kg indicating that the SSCP is more sensitive than the single dose mouse test for the detection of resistance to diminazene. However, none of the strains used in this study showed a sensitive SSCP profile while they were resistant in the single dose mouse test. The sequencing of the TcoAT1 gene of two sensitive, two intermediate and two resistant strains allowed the set up of a PCR-RFLP test for the discrimination between sensitive and resistant strains confirming the SSCP results for the 26 strains of this study.

Animals↗

Human P40/IL-9. Expression in activated CD4+ T cells, genomic organization, and comparison with the mouse gene.

P40 is a cytokine that was originally identified in the mouse as a T cell growth factor, but whose spectrum of potential targets was recently shown to include mast cells as well as megakaryoblastic leukemic cells. Given these multiple activities, it was proposed that the protein be renamed IL-9. The analysis of P40 genomic clones reported here shows that the human and mouse P40 genes consist of 5 exons spread over approximately 4 kb of DNA and organized in a similar fashion in both species. The two genes exhibit a high degree of identity in the coding sequence and in the 5' untranslated regions, which contain, among other consensus motifs, a conserved sequence for the binding of AP-1. Expression of human P40 was studied in PBMC. Treatment of the cells with PMA and a calcium ionophore induced strong expression of a 0.7-kb P40 mRNA. No message was detected in unstimulated cells or in cells stimulated with LPS or Staphylococcus aureus, indicating that P40 expression is not constitutive and suggesting that the gene is not easily activated in B lymphocytes and in monocytes. By contrast, T cell mitogens such as PHA or anti-CD3 antibodies induced a substantial P40 expression that was further enhanced in the presence of PMA. Cell fractionation experiments indicated that, under these conditions, the protein is preferentially induced in CD4+ T cells. The induction of P40 by anti-CD3 antibodies suggests that P40 production is part of the normal T cell response to antigenic stimulation.

Amino Acid Sequence↗

Cloning and expression of a cDNA for the human homolog of mouse T cell and mast cell growth factor P40.

A cDNA encoding the human homolog of mouse T-cell and mast cell growth factor P40 was derived from peripheral blood mononuclear cells (PBMC) stimulated with phytohemagglutinin and phorbol myristate acetate. Sequence analysis of the cDNA predicted a precursor protein of 144 amino acids including a signal peptide of 18 residues, a structure identical with that of mouse P40. The homology between the mouse and human proteins is 55% with a perfect conservation of the 10 cysteine residues present in the mature polypeptide. Expression of the cDNA for human P40 in a baculovirus vector yielded a protein capable of enhancing in vitro survival of human T cell lines.

Amino Acid Sequence↗

Cloning and characterization of a cDNA for a new mouse T cell growth factor (P40).

Recently, we described a murine helper T cell-derived molecule with T cell growth factor activity that is functionally and structurally distinct from IL-2, IL-4, and other known growth factors. This molecule, designated P40, was identified as a glycoprotein capable of supporting antigen-independent growth of certain helper T cell clones. Here, we report the cloning and expression of a cDNA for this new growth factor. The predicted mature protein is a cationic cysteine-rich polypeptide of 14 kD without significant homology to previously sequenced proteins.

Amino Acid Sequence↗

Cellular localization of stable solid liposomes in the liver of rats.

Small solid liposomes made from distearoylphosphatidyl choline and cholesterol (molar ratio 2:1) showed significant stability in plasma, with a half-life of about 24 hr after intravenous injection in rats. The major cellular uptake of intact liposomes was found in the liver and spleen, peaking after 2-4 hr in the liver and after 24 hr in the spleen. Isolation of parenchymal and non-parenchymal cells from rat livers at various intervals after injection of liposomes showed that both cell types adsorbed liposomal membranes and took up the liposomal contents. Our study has shown that most of the liposomal markers found in the liver shortly (less than 40 min) after administration stemmed from the liposomes adsorbed to extracellular binding sites, and that uptake into the cells took place subsequently. In non-parenchymal cells, uptake was rapid and the intracellular level remained rather constant after 40 min and for up to 4 hr. The uptake of liposomes by parenchymal cells was slower, it showed a lag-phase of approx. 1/2 hr and peaked at 2 hr, whereupon the radioactivity in parenchymal cells dropped. The contents of liposomes behaved in a manner similar to the membranes. It is concluded that, in addition to a rapid uptake of liposomes in non-parenchymal liver cells, there is a significant degree of association with parenchymal cells, provided that the liposomes administered are small (less than 100 nm in diameter) and stable.

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