Comparative effects of hypoxia and hypoxia mimetic cobalt chloride on in vitro adhesion, biofilm formation and susceptibility to amphotericin B of Candida glabrata

Gupta, Payal; Nath, Surendra; Meena, Ramesh Chandra; Kumar, Neha

doi:10.1016/j.mycmed.2014.08.003

Cited by 11 publications

(9 citation statements)

References 45 publications

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“…For quantification of biofilm formed at different time intervals (2, 4, 8, 10, 12, 24, and 48 h), XTT reduction assay was used. 80…”

Section: Methodsmentioning

confidence: 99%

“…After incubation, PBS was replaced with RPMI, and MTP was again incubated at 37 °C. For quantification of biofilm formed at different time intervals (2, 4, 8, 10, 12, 24, and 48 h), XTT reduction assay was used …”

Section: Methodsmentioning

confidence: 99%

“…For quantification of biofilm formed at different time intervals (2, 4, 8, 10, 12, 24, and 48 h), XTT reduction assay was used. 80 For studying the effect of CIN and EUG on C. glabrata and its clinical isolate biofilm, log phase cells were diluted in sterile PBS to a concentration of 1 × 10 7 cells mL −1 , and 100 μL of suspension was added into each well of MTP. 81 Plates were then incubated at 37 °C for 90 min (adhesion phase), and 200 μL of RPMI containing CIN and EUG in the concentration range of 512−2 μg mL −1 was added after washing wells thrice with PBS.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

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Effectiveness of Phytoactive Molecules on Transcriptional Expression, Biofilm Matrix, and Cell Wall Components of Candida glabrata and Its Clinical Isolates

Gupta

Sharma

et al. 2018

ACS Omega

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Toxicity challenges by antifungal arsenals and emergence of multidrug resistance scenario has posed a serious threat to global community. To cope up with this alarming situation, phytoactive molecules are richest, safest, and most effective source of broad spectrum antimicrobial compounds. In the present investigation, six phytoactive molecules [cinnamaldehyde (CIN), epigallocatechin, vanillin, eugenol (EUG), furanone, and epigallocatechin gallate] were studied against Candida glabrata and its clinical isolates. Among these, CIN and EUG which are active components of cinnamon and clove essential oils, respectively, exhibited maximum inhibition against planktonic growth of C. glabrata at a concentration of 64 and 128 μg mL –1 , respectively. These two molecules effectively inhibited and eradicated approximately 80% biofilm of C. glabrata and its clinical isolates from biomaterials. CIN and EUG increased reactive oxygen species generation, cell lysis, and ergosterol content in plasma membrane and reduced virulence attributes (phospholipase and proteinase) as well as catalase activity of C. glabrata cells. Reduction of mitochondrial membrane potential with increased release of cytochrome c from mitochondria to cytosol indicated initiation of early apoptosis in CIN- and EUG-treated C. glabrata cells. Transcriptional analysis showed that multidrug transporter ( CDR1 ) and ergosterol biosynthesis genes were downregulated in the presence of CIN, while getting upregulated in EUG-treated cells. Interestingly, genes such as 1,3-β-glucan synthase ( FKS1 ), GPI-anchored protein ( KRE1 ), and sterol importer ( AUS1 ) were downregulated upon treatment of CIN/EUG. These results provided molecular-level insights about the antifungal mechanism of CIN and EUG against C. glabrata including its resistant clinical isolate. The current data established that CIN and EUG can be potentially formulated in new antifungal strategies.

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“…For quantification of biofilm formed at different time intervals (2, 4, 8, 10, 12, 24, and 48 h), XTT reduction assay was used. 80…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: ■ Materials and Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Effectiveness of Phytoactive Molecules on Transcriptional Expression, Biofilm Matrix, and Cell Wall Components of Candida glabrata and Its Clinical Isolates

Gupta

Sharma

et al. 2018

ACS Omega

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“…Our previous study reported that CoCl 2 could stabilize hypoxia-inducible factor-1 alpha (HIF-1 α ), a key transcription factor for hypoxic condition in HPDLs [ 50 ]. In contrast to some lines of evidence, there were some aspects of detailed different mechanisms between hypoxia and CoCl 2 [ 51 , 52 ]. Thus, the results from the present study may not be directly implied to those of physical hypoxia setting.…”

Section: Discussionmentioning

confidence: 97%

Intermittent Compressive Stress Enhanced Insulin-Like Growth Factor-1 Expression in Human Periodontal Ligament Cells

Pumklin

Manokawinchoke

Bhalang

et al. 2015

International Journal of Cell Biology

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Mechanical force was shown to promote IGF-1 expression in periodontal ligament both in vitro and in vivo. Though the mechanism of this effect has not yet been proved, here we investigated the molecular mechanism of intermittent mechanical stress on IGF-1 expression. In addition, the role of hypoxia on the intermittent compressive stress on IGF-1 expression was also examined. In this study, human periodontal ligament cells (HPDLs) were stimulated with intermittent mechanical stress for 24 hours. IGF-1 expression was examined by real-time polymerase chain reaction. Chemical inhibitors were used to determine molecular mechanisms of these effects. For hypoxic mimic condition, the CoCl2 supplementation was employed. The results showed that intermittent mechanical stress dramatically increased IGF-1 expression at 24 h. The pretreatment with TGF-β receptor I or TGF-β1 antibody could inhibit the intermittent mechanical stress-induced IGF-1 expression. Moreover, the upregulation of TGF-β1 proteins was detected in intermittent mechanical stress treated group. Correspondingly, the IGF-1 expression was upregulated upon being treated with recombinant human TGF-β1. Further, the hypoxic mimic condition attenuated the intermittent mechanical stress and rhTGF-β1-induced IGF-1 expression. In summary, this study suggests intermittent mechanical stress-induced IGF-1 expression in HPDLs through TGF-β1 and this phenomenon could be inhibited in hypoxic mimic condition.

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“…Adaptations to hypoxia are required for the survival and enhanced virulence of pathogenic fungi such as C. albicans, C. glabrata, Cryptococcus neoformans and Aspergillus fumigatus (Grahl et al 2012;Gupta et al 2014Gupta et al , 2015. The function of most of the genes analyzed in this study is still unknown in C. glabrata.…”

Section: Discussionmentioning

confidence: 99%

Functional analysis of selected deletion mutants in Candida glabrata under hypoxia

2017

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Increased drug resistance in Candida glabrata (a model non-albicans Candida) calls for the identification of potential molecular targets for the development of effective drugs. Hypoxia (a state of low oxygen) is an important host factor, which affects the virulence of the pathogen and efficacy of drugs. In the present study, in vitro characterization of 13 null mutants of C. glabrata were done under hypoxic condition (1% O). These mutants have a major role to play in cellular pathways, viability and pathogenesis (cell wall biosynthesis, ergosterol synthesis, calcium-calcineurin, etc.). The in vitro growth, biofilm formation and susceptibility of biofilm to antifungal drugs of these mutants were compared with the control. Hypoxia reduced the susceptibility of planktonic cells to fluconazole. The mutants ecm33Δ, kre1Δ, rox1Δ, and kre2Δ showed maximum reductions in their biofilm activities (>20%). The selected mutants (upc2BΔ, kre2 Δ, ecm7Δ, rox1 Δ, mid1Δ, ecm33Δ, cch1Δ, kre1Δ) showed reduced biofilm activities (>30%) in the presence of 16 μg ml fluconazole under hypoxia. Functional analysis revealed that Kre1, Ecm33, Upc2B, Kre2, Ecm7, Cch1, Mid1 and Rox1 can be explored as a potential drug target for developing novel antifungal drugs.

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Comparative effects of hypoxia and hypoxia mimetic cobalt chloride on in vitro adhesion, biofilm formation and susceptibility to amphotericin B of Candida glabrata

Cited by 11 publications

References 45 publications

Effectiveness of Phytoactive Molecules on Transcriptional Expression, Biofilm Matrix, and Cell Wall Components of Candida glabrata and Its Clinical Isolates

Effectiveness of Phytoactive Molecules on Transcriptional Expression, Biofilm Matrix, and Cell Wall Components of Candida glabrata and Its Clinical Isolates

Intermittent Compressive Stress Enhanced Insulin-Like Growth Factor-1 Expression in Human Periodontal Ligament Cells

Functional analysis of selected deletion mutants in Candida glabrata under hypoxia

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