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1.
BMC Res Notes ; 17(1): 2, 2024 Jan 02.
Artículo en Inglés | MEDLINE | ID: mdl-38167002

RESUMEN

OBJECTIVES: Candida albicans, a polymorphic yeast, is one of the most common, opportunistic fungal pathogens of humans. Among the different morphological forms, opaque form is one of the least-studied ones. This opaque phenotype is essential for mating and is also reported to be involved in colonizing the gastrointestinal tract. Considering the significance of the clinical and sexual reproduction of C. albicans, we have investigated the morphophysiological modulations in opaque form using a proteomic approach. DATA DESCRIPTION: In the current investigation, we have used Micro-Liquid Chromatography-Mass Spectrometry (LC-MS/MS) analysis to create a protein profile for opaque-specific proteins. Whole-cell proteins from C. albicans (ATCC10231) cells that had been cultured for seven days on synthetic complete dextrose (SCD) medium in both as an opaque (test) and as a white (control) form cells were extracted, digested, and identified using LC-MS/MS. This information is meant to serve the scientific community and represents the proteome profile (SWATH Spectral Libraries) of C. albicans opaque form.


Asunto(s)
Candida albicans , Proteínas Fúngicas , Humanos , Candida albicans/genética , Proteínas Fúngicas/metabolismo , Proteoma/metabolismo , Cromatografía Liquida , Proteómica , Genes del Tipo Sexual de los Hongos , Espectrometría de Masas en Tándem , Fenotipo , Regulación Fúngica de la Expresión Génica
2.
Arch Microbiol ; 203(4): 1439-1450, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33392625

RESUMEN

Chlamydomonas reinhardtii is one of the most characterized green algae. The open-pond cultivation can be challenging due to sensitivity of strain to fluctuating environmental conditions and unavailability of low-cost photoautotrophic media. In this study, the photoautotrophic growth of C. reinhardtii was evaluated in 1-m2 open ponds placed in greenhouse. Sodium bicarbonate (NaHCO3) was evaluated as an alternative buffering agent to tris. The effect of buffer and pH was tested. The growth was studied in the presence of various nitrogen [urea and ammonium bicarbonate (NH4HCO3)] sources. In the study, it was found that 125-ppm NaHCO3 as an optimum concentration. The buffering agent in the media was found to have major impact on growth. Without buffering agent, culture did not grow, and pH drop was observed. The sodium bicarbonate-buffered media reported to have the lowest bacterial contamination (18.3%), highest AFDW per OD (0.39 ± 0.027 g/L) and higher Fv/Fm (0.714 ± 0.016), whereas these values were found to be 62%, 0.19 ± 0.02 g/L and 0.537 ± 0.053 for tris-grown culture, respectively. The pH 7.0-7.5 was determined as an optimum, whereas pH 6.5-7.0 and 8.0-8.5 were found to affect the growth and induce palmelloidy. The OD and AFDW of culture grown in NH4HCO3 were found equivalent to a standard nitrogen source (NH4Cl), whereas culture shown poor growth in urea. Based on these data, NH4HCO3 media recipe and the optimized cultivation parameters were selected for photoautotrophic cultivation of Chlamydomonas in greenhouse open ponds.


Asunto(s)
Procesos Autotróficos , Chlamydomonas reinhardtii/crecimiento & desarrollo , Procesos Fototróficos , Estanques/química , Chlamydomonas reinhardtii/metabolismo , Medios de Cultivo/química , Concentración de Iones de Hidrógeno , Nitrógeno/análisis , Bicarbonato de Sodio/análisis
3.
J Proteomics ; 185: 25-38, 2018 08 15.
Artículo en Inglés | MEDLINE | ID: mdl-29959084

RESUMEN

To understand the impact of fetal bovine serum (FBS) on metabolism and cellular architecture in addition to morphogenesis, we have identified FBS responsive proteome of Candida albicans. FBS induced 34% hyphae and 60% pseudohyphae in C. albicans at 30 °C while 98% hyphae at 37 °C. LC-MS/MS analysis revealed that 285 proteins modulated significantly in response to FBS at 30 °C and 37 °C. Out of which 152 were upregulated and 62 were downregulated at 30 °C while 18 were up and 53 were downregulated at 37 °C. Functional annotation suggests that FBS may inhibit glycolysis and fermentative pathway and enhance oxidative phosphorylation (OxPhos), TCA cycle, amino acid and fatty acid metabolism indicating a use of alternative energy source by C. albicans. OxPhos inhibition assay using sodium azide corroborated the correlation between inhibition of glycolysis and enhanced OxPhos with pseudohyphae formation. C. albicans induced hyphae in response to FBS irrespective of down regulation of Ras1,Asr1/Asr2, indicates the possible involvement of MAPK and cAMP-PKA independent pathway. The Cell wall of cells grown in presence of FBS at 30 °C was rich in mannan, Beta 1,3-glucan and chitin while membranes were rich in ergosterol compared to those grown at 37 °C. SIGNIFICANCE OF THE STUDY: This is the first study suggesting a correlation between OxPhos and morphogenesis especially pseudohyphae formation in C. albicans. Our data also indicate that fetal bovine serum (FBS) induced morphogenesis is multifactorial and may involve MAPK and cAMP-PKA independent pathway. In addition to morphogenesis, our study provides an insight in to the modulation of metabolism and cellular architecture of C. albicans in response to FBS.


Asunto(s)
Candida albicans/crecimiento & desarrollo , Candida albicans/metabolismo , Morfogénesis/fisiología , Fosforilación Oxidativa , Proteoma/metabolismo , Suero/fisiología , Animales , Bovinos , Cromatografía Liquida , Proteínas Fúngicas/análisis , Proteínas Fúngicas/metabolismo , Hifa/crecimiento & desarrollo , Hifa/metabolismo , Espectrometría de Masas en Tándem
4.
Med Mycol ; 56(5): 565-578, 2018 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-29420815

RESUMEN

Anti-Candida potential of limonene was evaluated against planktonic growth, biofilm (adhesion, development and maturation) and morphogenesis of Candida albicans in this study. Limonene is a major constituent of citrus oil and most frequently used terpene in food and beverage industry due to its pleasant fragrance, nontoxic, and is generally recognized as safe (GRAS) flavoring agent as well as treatment option in many gastrointestinal diseases.Limonene exhibited excellent anti-Candida activity and was equally effective against planktonic growth of C. albicans isolates differentially susceptible to FLC (N = 35). Limonene inhibited morphogenesis significantly at low concentration. However, it showed stage dependent activity against biofilm formation, that is, it was more effective against adhesion followed by development and maturation. Limonene also exhibited excellent synergy with FLC against planktonic and biofilm growth. SWATH-MS analysis led to identification of limonene responsive proteins that provided molecular insight of its anti-Candida activity. Proteomic analysis revealed upregulation of proteins involved in cell wall glucan synthesis (Kre6); oxidative stress (Rhr2, Adh7 and Ebp1); DNA damage stress (Mbf1 and Npl3); nucleolar stress (Rpl11, Rpl7, Rpl29, Rpl15) and down regulation of cytoskeleton organization (Crn1, Pin3, Cct8, Rbl2), and so forth, in response to limonene. Limonene mediated down regulation of Tps3 indicates activation of caspase (CaMca1) and induction of apoptosis in C. albicans. These results suggest that limonene inhibits C. albicans growth by cell wall/membrane damage induced oxidative stress that leads to DNA damage resulting into modulation of cell cycle and induction of apoptosis through nucleolar stress and metacaspase dependent pathway.


Asunto(s)
Apoptosis/efectos de los fármacos , Biopelículas/efectos de los fármacos , Candida albicans/efectos de los fármacos , Ciclohexenos/farmacología , Terpenos/farmacología , Antifúngicos/farmacología , Biopelículas/crecimiento & desarrollo , Candida albicans/crecimiento & desarrollo , Candida albicans/metabolismo , Cromatografía Liquida , Farmacorresistencia Fúngica/efectos de los fármacos , Sinergismo Farmacológico , Fluconazol/farmacología , Proteínas Fúngicas/genética , Puntos de Control de la Fase G1 del Ciclo Celular/efectos de los fármacos , Regulación Fúngica de la Expresión Génica/efectos de los fármacos , Limoneno , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana/efectos de los fármacos , Proteómica , Espectrometría de Masas en Tándem
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