Our PSC-AgNP preparation makes for a promising antifungal agent that can downregulate isocitrate lyase. © 2017 Society of Chemical Industry.
The virulence of Candida albicans is dependent upon fitness attributes as well as virulence factors. These attributes include robust stress responses and metabolic flexibility. The assimilation of carbon sources is important for growth and essential for the establishment of infections by C. albicans. Previous studies showed that the C. albicans ICL1 genes, which encode the glyoxylate cycle enzymes isocitratelyase are required for growth on non-fermentable carbon sources such as lactate and oleic acid and were repressed by 2% glucose. In contrast to S. cerevsiae, the enzyme CaIcl1 was not destabilised by glucose, resulting with its metabolite remaining at high levels. Further glucose addition has caused CaIcl1 to lose its signal and mechanisms that trigger destabilization in response to glucose. Another purpose of this study was to test the stability of the Icl1 enzyme in response to the dietary sugars, fructose, and galactose. In the present study, the ICL1 mRNAs expression was quantified using Quantitative Real Time PCR, whereby the stability of protein was measured and quantified using Western blot and phosphoimager, and the replacing and cloning of ICL1 ORF by gene recombination and ubiquitin binding was conducted via co-immuno-precipitation. Following an analogous experimental approach, the analysis was repeated using S. cerevisiaeas a control. Both galactose and fructose were found to trigger the degradation of the ICL1 transcript in C. albicans. The Icl1 enzyme was stable following galactose addition but was degraded in response to fructose. C. albicans Icl1 (CaIcl1) was also subjected to fructose-accelerated degradation when expressed in S. cerevisiae, indicating that, although it lacks a ubiquitination site, CaIcl1 is sensitive to fructose-accelerated protein degradation. The addition of an ubiquitination site to CaIcl1 resulted in this enzyme becoming sensitive to galactose-accelerated degradation and increases its rate of degradation in the presence of fructose. It can be concluded that ubiquitin-independent pathways of fructose-accelerated enzyme degradation exist in C. albicans.
Background: A major characteristic of Candida biofilm cells that differentiates them from free-floating cells is their high tolerance to antifungal drugs. This high resistance is attributed to particular biofilm properties, including the accumulation of extrapolymeric substances, morphogenetic switching, and metabolic flexibility. Objectives: This study evaluated the roles of metabolic processes (in particular the glyoxylate cycle) on biofilm formation, antifungal drug resistance, morphology, and cell wall components. Methods: Growth, adhesion, biofilm formation, and cell wall carbohydrate composition were quantified for isogenic Candida albicans ICL1/ICL1, ICL1/icl1, and icl1/icl1 strains. The morphology and topography of these strains were compared by light microscopy and scanning electron microscopy. FKS1 (glucan synthase), ERG11 (14-α-demethylase), and CDR2 (efflux pump) mRNA levels were quantified using qRT-PCR. Results: The ICL1/icl1 and icl1/icl1 strains formed similar biofilms and exhibited analogous drug-tolerance levels to the control
MAP, an open access journal extracts from Aquilaria crassna stem bark against panel of cancer cell lines; (ii) to study the antiangiogenic effects of Aquilaria crassna most active extract. Materials and Methods Chemicals and reagents All cell cultures and their reagents were purchased from Gibco (USA). Phosphate buffered saline, trypsin, HIFBS, PS, fibrinogen, aprotinin, thrombin, suramin, aprotinin, 6-aminocaproic acid, L-glutamine, thrombin, and gentamicin were purchased from Sigma (Germany). MTT (3-(4,5-dimethylthiazol-2-yl)-2,5diphenyl tetrazolium bromide) were procured from Sigma-Aldrich (USA). Dimethyl sulfoxide (DMSO) was purchased from Fluka (USA). Cell lines and culture conditions The HUVEC cell line (catalogue number, C2517A), human colorectal carcinoma cell line HCT-116 (catalogue number CCL-247), human hormone sensitive invasive breast cancer cell line MCF-7 (catalogue number CRL-1469), Human pancreatic carcinoma cell line (PANC-1) catalogue number (HTB-14) and human prostate cancer cell line PC-3 (catalog number CRL-1435) were purchased from ScienCell (USA). HUVECs were maintained in ECM (ScienCell) supplemented with endothelial cell growth supplements (ECGS), 5% HIFBS, and 1% PS. HCT-116 cells and PANC-1 were maintained in RPMI, whereas MCF-7 and PC-3 cells were maintained in DMEM and F12K medium, respectively. The media were supplemented with 5% HIFBS and 1% PS. Cells were cultured in a humidified incubator at 37°C supplied by 5% CO 2. Cell culture work was conducted in sterile conditions using a Class II biosafety cabinet (ESCO, USA).
Background: Candida glabrata is a ubiquitous organism and this species is one of the normal floras that commonly found in oral cavity, gastrointestinal and vaginal tract. However, they can be pathogenic and caused serious infection that more likely to occur in immunocompromised patients. Widespread and increased usage of immunosuppressive therapies with the additional of broad-spectrum antibiotic treatment can also lead to the significant increment of Candida infections. Candida species can cause nosocomial infection that associated with alarming high mortality rate worldwide. C. glabrata is one of the Candida species that raise an attention of a researcher due to their high resistance to certain drugs and antifungal as well as host immune system.Objective: To investigate certain aspects of the C. glabrata that contributes to their resistance to drugs and as host in immune system.Results: Microorganisms including Candida have an ability to evolve in order to sustain their survival and it is creating a new microbial community that are more resistant. C. glabrata possessed several strategies for their resistance including production of biofilm, genomic plasticity as well as chromatin alteration. On the other hand, their resistance may cause by the mutation in certain gene. Besides, they also use a variety of mechanisms that enable their adaptation works successfully and their survival as host in immune system. Conclusion:A significant number of researches have been conducted to elucidate resistant mechanism of C. glabrata thus, much information has been gained on how their resistant mechanism works and has enhanced our knowledge on the virulence and resistance mechanisms associated with Candida species especially C. glabrata. Further studies and investigation in this area are crucial particularly on the identification and development of new targets for novel therapeutics against these recently emerged pathogens.
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