Two isolates representing a new species of Scheffersomyces were isolated from rotting wood samples collected in an Amazonian forest ecosystem in Brazil. Analysis of the sequences of the D1/D2 domains showed that this new species is phylogenetically related to Scheffersomyces NYMU 15730, a species without a formal description, and the two are in an early emerging position with respect to the xylose-fermenting subclade containing Scheffersomyces titanus and Scheffersomyces stipitis. Phylogenomic analyses using 474 orthologous genes placed the new species in an intermediary position between Scheffersomyces species and the larger genus Spathaspora and the Candida albicans/Lodderomyces clade. The novel species, Scheffersomyces stambukii f.a., sp. nov., is proposed to accommodate these isolates. The type strain of Scheffersomyces stambukii sp. nov. is UFMG-CM-Y427 (=CBS 14217). The MycoBank number is MB 824093. In addition, we studied the xylose metabolism of this new species.
During studies of the yeast diversity associated with rotting wood in Brazil and fruits, plants and insects in French Guiana, three strains of a new species were isolated. Analysis of the sequences of the internal transcribed spacer (ITS)-5.8S and D1/D2 domains of the large subunit of the rRNA gene showed that this species belongs to the genus Hyphopichia and its closest relative is Candida homilentoma. These species differ by 44 nucleotide substitutions in D1/D2 sequences. A new species Hyphopichia buzzinii f. a., sp. nov., is proposed to accommodate these isolates. The type strain of Hyphopichia buzzinii sp. nov. is CLIB 1739 (=CBS 14300 = UFMG-CM-Y6121; MycoBank number is MB 815609). In addition, we isolated 11 strains of C. homilentoma from rotting wood, leaf surfaces, and water bodies in Brazil, and these strains when crossed among one another and with the type strain (CBS 6312) of this species, produced hat-shaped ascospores typical of the genus Hyphopichia. We describe the teleomorph of C. homilentoma as a new combination, Hyphopichia homilentoma comb. nov. (type strain CBS 6312; MycoBank number is MB 820009). We also propose to transfer the other six Candida species of the Hyphopichia clade to this genus as new combinations. Hyphopichia homilentoma produced ethanol and xylitol from D-xylose whereas H. buzzinii was only able to convert this pentose to xylitol.
Conclusion:This study provides evidence that Ang-(1-5) is an endogenous AT 2 R agonist with high efficacy towards the AT 2 R. The early signaling phosphorylation pattern resembles those of other protective RAS agonists, such as C21 and Ang-(1-7).
Introduction: Recombinant human ACE2 increases the circulating levels of angiotensin-(1-5) [Ang-(1-5)], a peptide thus far regarded as biologically inactive. Since ACE2 is a central component of the protective RAS, we hypothesized that Ang-(1-5) is a new biologically active peptide within this hormonal system. Objective: To investigate biological activity and signaling mechanisms of Ang-(1-5). Methods: In order to show a biological effect and to test whether Ang-(1-5) signals through the AT 2 -receptor (AT 2 R), nitric oxide (NO) release was measured by DAF-FM fluorescence in AT 2 R transfected (AT 2 R-CHO) or non-transfected (NT-CHO) CHO cells, treated with Ang-(1-5) or C21 (AT 2 R agonist, positive control) (0.1nM to 10μM) for 15 minutes. Vehicle (cell media) treated cells served as negative control. To investigate Ang-(1-5) signaling patterns, human aortic endothelial cells (HAEC) were treated with vehicle or Ang-(1-5) (1μM) for 1, 3, 5 or 20 minutes. Proteins were harvested, digested and labeled with TMTpro-16plex. Phosphopeptide enrichment was carried out by TiO 2 . Samples were subjected to LC-MS/MS analysis and the resulting mass spectra were searched against the human SwissProt database. Results: Ang-(1-5) induced a concentration-dependent increase in NO production in AT 2 -CHO cells (E max : 65.60 ± 14.02%), thus proving its biological activity. Ang-(1-5) had 69% higher efficacy than the established AT 2 R agonist C21 (E max : 38.76 ± 10.24%). Ang-(1-5) seems to signal through the AT 2 R, because effects on NO release were absent in NT-CHO cells. Treatment of HAEC with Ang-(1-5) significantly modified the phosphorylation status of 831 proteins at 1799 residues. The majority of residues (1079) were dephosphorylated while 729 residues were phosphorylated. Changes in protein phosphorylation in response to Ang-(1-5) occurred at all investigated time points, most of them after 20 minutes. Functional bioinformatic analysis revealed a cluster of proteins involved in cell cycle and cell division regulation. Conclusion: This study provides evidence that Ang-(1-5) is an endogenous AT 2 R agonist with high efficacy towards the AT 2 R. The early signaling phosphorylation pattern resembles those of other protective RAS agonists, such as C21 and Ang-(1-7).
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