New 1,4-anthracenediones bearing fused-heterocycle rings were synthesized and evaluated as cytotoxics, antifungals and antivirals. Some of them showed GI50 at the μM level.
Identification of filamentous fungi by conventional phenotypic methods are time-consuming, and a correct identification at the species level is prone to errors. Therefore, a more accurate and faster time-to-results, and cost-effective technique, is required, such as the Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS). In this study, we describe the development of an in-house spectra library for the identification of filamentous fungi frequently isolated from patients with infections. An in-house spectra library was constructed using 14 reference strains grown in solid medium. Clinical isolates were identified either by the in-house spectra library or the Biotyper commercial library from Bruker Daltonics. Fungal identification was carried following the Biotyper’s established scores: ≤1.699: not reliably identified (NRI); 1.700–1.999: genus-level; ≥2.000: species-level. Clinical isolates were identified, with the in-house library, at species- and genus-level at 88.70% (55) and 3.22% (2), respectively. While 4.80% (3) was NRI and 3.22% (2) was discrepant concerning sequencing. On the contrary, identification up to species and genus-level with the commercial library was 44.44% (16) and 22.22% (8), respectively. NRI and the discrepancy was 30.55% (11) and 2.77% (1), respectively. For the reaming 26 isolates, 16 from Neoscytalidium dimidiatum and 10 from Sporothrix spp., respectively, the absence of spectrum and the specific spectra within the Sporothrix complex in the commercial library resulted in the inability to obtain an identification. In conclusion, the current results advocate the importance that each clinical microbiological laboratory needs to develop an ad hoc library associated with the MALDI-TOF MS fungal identification to overcome the limitations of the available commercial libraries.
Multi-drug resistant species such as Candida auris are a global health threat. This scenario has highlighted the need to search for antifungal alternatives. Essential oils (EOs), or some of their major compounds, could be a source of new antifungal molecules. The aim of this study was to evaluate the in vitro activity of EOs and some terpenes against C. auris and other Candida spp. The eleven EOs evaluated were obtained by hydro-distillation from different Colombian plants and the terpenes were purchased. EO chemical compositions were obtained by gas chromatography/mass spectrometry (GC/MS). Antifungal activity was evaluated following the CLSI standard M27, 4th Edition. Cytotoxicity was tested on the HaCaT cell line and fungal growth kinetics were tested by time–kill assays. Candida spp. showed different susceptibility to antifungals and the activity of EOs and terpenes was strain-dependent. The Lippia origanoides (thymol + p-cymene) chemotype EO, thymol, carvacrol, and limonene were the most active, mainly against drug-resistant strains. The most active EOs and terpenes were also slightly cytotoxic on the HaCaT cells. The findings of this study suggest that some EOs and commercial terpenes can be a source for the development of new anti-Candida products and aid the identification of new antifungal targets or action mechanisms.
Within the Neoscytalidium genus, N. dimidiatum, N. oculus, N. orchidacearum, and N. novaehollandiae have been recognized. Although these species are frequently found in soil, N. dimidiatum has been identified as an etiologic agent of onychomycosis or dermatomycosis, and N. oculus has been identified as an etiologic agent of an ocular lesion. All these species can be cultured in vitro, but their morphological identification by macroscopic and microscopic traits is difficult and imprecise due to their similarity. In this study, 34 isolates of Neoscytalidium spp. from 32 onychomycosis and two dermatomycosis cases in Medellin (Colombia) were identified at the species level using sequencing of the ITS1+5.8S+ITS2 nuclear rDNA region and MALDI-TOF mass spectrometry (MS). Neoscytalidium dimidiatum strain MUM 17.21 was used to construct the reference spectrum in the in-house library to identify the clinical isolates by MALDI-TOF MS. Additionally, N. dimidiatum PPC-216 and PLAB-055 strains were used to validate the in-house constructed reference spectra. Although four groups were observed in the dendrogram obtained from the proteins of each isolate profile, MALDI-TOF MS and sequencing results are in accordance, since all isolates were identified as N. dimidiatum.
Sloanea is a plant genus, native to tropical regions, used in medicinal practices for its anti-inflammatory properties. This study aimed to determine the antioxidant activity, sun protective factor (SPF), and antifungal of extracts obtained from two species of Sloanea and to develop extract-based gels with antioxidants, photoprotective, and anti-Candida albicans effects. Ethanolic extracts from S. medusula and S. calva collected in Chocó, Colombia, were used for antioxidant activity and SPF determination using the DPPH assay and the Mansur equation, respectively. Extracts were characterized using HPLC-MS and used to prepare the gels. The viscosity of the extract-based gels was evaluated using an MCR92 rheometer. In addition, the anti-Candida activity of extracts against five yeasts and anti-C. albicans of gels were evaluated following the Clinical and Laboratory Standards Institute M27, 4th Edition. High DPPH radical scavenging activity (42.4% and 44.7%) and a high SPF value (32.5 and 35.4) were obtained for the extracts of S. medusula and S. calva, respectively. Similarly, extract-based gels showed significant DPPH radical scavenging activity of 54.5% and 53.0% and maximum SPF values of 60 and 57. Extract from S. medusula showed an important antifungal activity against C. albicans (minimal inhibitory concentration (MIC) of 2 µg/mL). In contrast, S. calva extract was active against C. krusei, C. albicans (MIC of 2 µg/mL) and C. tropicalis (MIC of 4 µg/mL). Sloanea medusula gel (0.15%) exhibited an important C. albicans growth inhibition (98%), while with S. calva gel (0.3%) growth inhibition was slightly lower (76%). Polyphenolic and triterpenoid compounds were tentatively identified for S. medusula and S. calva, respectively. Both extracts can be considered promising sources for developing photoprotective gels to treat skin infections caused by C. albicans.