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Candida molecular typing

Thierry Kammalac Ngouana, Pascal Drakulovski, Donika Krasteva, Rufin Kuipou Toghueo, Charles Kouanfack, Jacques Reynes, Eric Delaporte, Fabrice Fekam Boyom, Michèle Mallié, Sébastien Bertout
The molecular epidemiology and the antifungal susceptibility profiles of Candida albicans are scarce in Cameroon. Authors studied the genetic diversity and the antifungal susceptibility of C. albicans isolates from Yaoundé HIV-infected patients. Clinical isolates were obtained by mycological diagnosis of oropharyngeal swabs, stools, urine, and vaginal swabs from patients. C. albicans isolates were confirmed by the Light cycler real-time PCR of the ITS1 region of the 5.8s ribosomal DNA. The ABC genotypes and the Hwp1 gene amplification were carried out with specific primers...
October 14, 2016: Medical Mycology: Official Publication of the International Society for Human and Animal Mycology
Dhammika H M L P Navarathna, Ruvini U Pathirana, Michail S Lionakis, Kenneth W Nickerson, David D Roberts
Formation of chlamydospores by Candida albicans was an established medical diagnostic test to confirm candidiasis before the molecular era. However, the functional role and pathological relevance of this in vitro morphological transition to pathogenesis in vivo remain unclear. We compared the physical properties of in vitro-induced chlamydospores with those of large C. albicans cells purified by density gradient centrifugation from Candida-infected mouse kidneys. The morphological and physical properties of these cells in kidneys of mice infected intravenously with wild type C...
2016: PloS One
Rita Teixeira-Santos, Elisabete Ricardo, Ricardo J Branco, Maria M Azevedo, Acácio G Rodrigues, Cidália Pina-Vaz
Patients with multiple comorbidities are often administered simultaneously or sequentially antifungals and antibacterial agents, without full knowledge of the consequences of drug interactions. Considering the clinical relevance of liposomal amphotericin B (L-AMB), the association between L-AMB and six antibacterial agents was evaluated against four clinical isolates and one type strain of Candida spp. and two clinical isolates and one type strain of Aspergillus fumigatus. In order to evaluate such combined effects, the minimal inhibitory concentration (MIC) of L-AMB was determined in the presence of 0...
2016: Frontiers in Microbiology
Xiao-Fei Zhang, Guang-Yu Yang, Yong Zhang, Yuan Xie, Stephen G Withers, Yan Feng
The local flexibility of an enzyme's active center plays pivotal roles in catalysis, however, little is known about how the flexibility of these flexible residues affects stability. In this study, we proposed an active center stabilization (ACS) strategy to improve the kinetic thermostability of Candida rugosa lipase1. Based on the B-factor ranking at the region ~10 Å within the catalytic Ser209, 18 residues were selected for site-saturation mutagenesis. Based on three-tier high-throughput screening and ordered recombination mutagenesis, the mutant VarB3 (F344I/F434Y/F133Y/F121Y) was shown to be the most stable, with a 40-fold longer in half-life at 60 °C and a 12...
September 26, 2016: Scientific Reports
Henrique Marconi Sampaio Pinhati, Luiz Augusto Casulari, Ana Carolina Remondi Souza, Ricardo Andreotti Siqueira, Camila Maria Gomes Damasceno, Arnaldo Lopes Colombo
BACKGROUND: Candidemia is an increasing problem in tertiary care hospitals worldwide. Here, we report the first outbreak of candidemia caused by fluconazole-resistant C. parapsilosis (FRCP) strains in Brazil. METHODS: This was a cross-sectional study of clinical and microbiological data of all candidemic episodes diagnosed from July 2011 to February 2012 in a 200-bed tertiary care hospital. Initial yeast identification and susceptibility testing were performed using the VITEK 2 - System...
2016: BMC Infectious Diseases
Sébastien Imbert, Louise Castain, Antoine Pons, Stéphanie Jacob, Isabelle Meyer, Martine Palous, Corinne Vezinet, Olivier Langeron, Christophe Hennequin, Antoine Monsel, Arnaud Fekkar
OBJECTIVES: to give an indication of a fitness cost conferred by FKS mutation-associated echinocandin resistance in Candida glabrata during human infection. METHODS: six C. glabrata clinical strains sequentially isolated from blood and a hepatic abscess in a solid organ transplant recipient were analysed. The patient had received long-term azole and echinocandin therapy for invasive aspergillosis and persistent candidemia. Minimal inhibitory concentrations were determined by the EUCAST broth microdilution method...
July 30, 2016: Clinical Microbiology and Infection
A Martinez, I Cavello, G Garmendia, C Rufo, S Cavalitto, S Vero
Various microbial groups are well known to produce a range of extracellular enzymes and other secondary metabolites. However, the occurrence and importance of investment in such activities have received relatively limited attention in studies of Antarctic soil microbiota. Sixty-one yeasts strains were isolated from King George Island, Antarctica which were characterized physiologically and identified at the molecular level using the D1/D2 region of rDNA. Fifty-eight yeasts (belonging to the genera Cryptococcus, Leucosporidiella, Rhodotorula, Guehomyces, Candida, Metschnikowia and Debaryomyces) were screened for extracellular amylolytic, proteolytic, esterasic, pectinolytic, inulolytic xylanolytic and cellulolytic activities at low and moderate temperatures...
September 2016: Extremophiles: Life Under Extreme Conditions
Damien Balestrino, Mercédès Quintana, Nicolas Charbonnel, Christiane Forestier, Claire Lartigue, Bertrand Souweine
BACKGROUND AND OBJECTIVES: Interdialytic lock solutions should maintain catheter patency and prevent catheter infections. We aimed to determine in which conditions injectable anticoagulant agents (IAAs) combined with ethanol are compatible and to assess the antibiofilm activity of the selected combination and its effects on dialysis catheters (DC). METHODS: The solubility and compatibility of unfractionated heparin (UFH), low molecular weight heparins (LMWHs), heparinoids and fondaparinux (50 to 2,500 U/mL) in 30 to 70% ethanol were determined by visual observation...
2016: PloS One
Matthias Sipiczki, Ratna Mohd Tap
In the taxonomic study of yeasts isolated from flowers in Cagayan de Oro, Mindenao Island, The Philippines, strains were identified representing Kabatiella microsticta, Metschnikowia koreensis and a hitherto undescribed dimorphic species. The sequences of the D1/D2 domains of the large subunit (LSU) 26S rRNA genes, the internal transcribed spacer (ITS) regions and the small subunit (SSU) 18S rRNA genes were identical in the strains of the latter group and differed from the corresponding sequences of the type strain of the closest relative species, Candida duobushaemulonii, by 4% (D1/D2), 7% (ITS) and 1% (SSU)...
July 13, 2016: International Journal of Systematic and Evolutionary Microbiology
Filipe D S Gorza, Romário J da Silva, Tarquin F Trescher, Graciela C Pedro, Maria A Oliveira de Sousa, Paula C S Souto, Josmary R Silva, Nara C de Souza
The development of systems for the controlled release of drugs is important because they allow the control of drug absorption and tissue distribution and also can reduce local toxicity. This study aimed to assemble and characterize two types of release systems, consisting of layer-by-layer films obtained from poly(allylamine) hydrochloride with chlorophyll (PAH/CHL films) or chlorophyll incorporated into dipalmitoylphosphatidylcholine liposomes (PAH/Lip+CHL films). For these systems, the molecular aggregation, growth process, thermally stimulated desorption, wettability, and controlling release of CHL was studied by using UV-vis spectroscopy and wetting contact angle analysis...
September 2016: Photodiagnosis and Photodynamic Therapy
Yanjiao Ding, Yaoyao Li, Zhenyu Li, Juanli Zhang, Chunhua Lu, Haoxin Wang, Yuemao Shen, Liangcheng Du
BACKGROUND: Alteramide B (ATB), isolated from Lysobacter enzymogenes C3, was a new polycyclic tetramate macrolactam (PTM). ATB exhibited potent inhibitory activity against several yeasts, particularly Candida albicans SC5314, but its antifungal mechanism is unknown. METHODS: The structure of ATB was established by extensive spectroscopic analyses, including high-resolution mass spectrometry, 1D- and 2D-NMR, and CD spectra. Flow cytometry, fluorescence microscope, transmission electron microscope, molecular modeling, overexpression and site-directed mutation studies were employed to delineate the anti-Candida molecular mechanism of ATB...
October 2016: Biochimica et Biophysica Acta
Alieh Ameri, Ghadamali Khodarahmi, Farshid Hassanzadeh, Hamid Forootanfar, Gholam-Hosein Hakimelahi
The present study was planned to design some novel aldimine-type Schiff bases bearing 3,4,5-trimethoxyphenyl and 1,2,4-triazole-3-thione/thiol as potential tubulin polymerization inhibitors. The obtained results of the molecular docking study using the tubulin complex (PDB code: 1SA0) showed that compounds H-25 and H-26 were well fitted in the colchicine binding site of tubulin with binding energies of -8.68 and -8.40 kcal/mol, respectively, in comparison to the main ligand (-8.20 kcal/mol). In parallel, molecular simulations were also performed on five other 3,4,5-trimethoxyphenyl-containing ligand targets including hsp90, VEGFR2, and human and microbial (Staphylococcus aureus and Candida albicans) dihydrofolate reductase, among which H-17, H-45, H-27, H-02, and H-19 were the most suitable compounds, respectively...
August 2016: Archiv der Pharmazie
Ho Shin Kim, Doyoung Eom, Yoon-Mo Koo, Yaroslava G Yingling
In order to understand how cations affect the structural changes and enzyme activity of Lipase B from Candida antarctica, we performed all-atom molecular dynamics simulations of CALB in four types of ionic liquids (ILs) with varying sizes of imidazolium cations and correlated these results with the experimentally determined CALB activity. The imidazolium cations under study differ in the alkyl tail length in the following order: [Emim](+) < [Bmim](+) < [Hmim](+) < [Omim](+). We observed that the best enzyme activity and structural stability of CALB are obtained in [Bmim][TfO] and [Hmim][TfO]...
August 10, 2016: Physical Chemistry Chemical Physics: PCCP
Luis A Vale-Silva, Beat Moeckli, Riccardo Torelli, Brunella Posteraro, Maurizio Sanguinetti, Dominique Sanglard
Candida glabrata is the second most common Candida species causing disseminated infection, after C. albicans. C. glabrata is intrinsically less susceptible to the widely used azole antifungal drugs and quickly develops secondary resistance. Resistance typically relies on drug efflux with transporters regulated by the transcription factor Pdr1. Gain-of-function (GOF) mutations in PDR1 lead to a hyperactive state and thus efflux transporter upregulation. Our laboratory has characterized a collection of C. glabrata clinical isolates in which azole resistance was found to correlate with increased virulence in vivo...
March 2016: MSphere
Masaaki Konishi, Manako Fujita, Yu Ishibane, Yuki Shimizu, Yusuke Tsukiyama, Masashi Ishida
Eleven biosurfactant-producing strains were newly isolated from environmental samples using a drop-collapse assay and thin-layer chromatography (TLC). According to the TLC analysis, the separation patterns of the glycolipid spots of nine dominant strains corresponded to that of the sophorolipids produced by a Starmerella bombicola type strain. The retention factor values of the spot patterns of two strains were less than those of the others. Two representative major products were purified, and their molecular structures were determined...
October 2016: Bioscience, Biotechnology, and Biochemistry
Robert Krause, Bettina Halwachs, Gerhard G Thallinger, Ingeborg Klymiuk, Gregor Gorkiewicz, Martin Hoenigl, Jürgen Prattes, Thomas Valentin, Katharina Heidrich, Walter Buzina, Helmut J F Salzer, Jasmin Rabensteiner, Florian Prüller, Reinhard B Raggam, Andreas Meinitzer, Christine Moissl-Eichinger, Christoph Högenauer, Franz Quehenberger, Karl Kashofer, Ines Zollner-Schwetz
Whether the presence of Candida spp. in lower respiratory tract (LRT) secretions is a marker of underlying disease, intensive care unit (ICU) treatment and antibiotic therapy or contributes to poor clinical outcome is unclear. We investigated healthy controls, patients with proposed risk factors for Candida growth in LRT (antibiotic therapy, ICU treatment with and without antibiotic therapy), ICU patients with pneumonia and antibiotic therapy and candidemic patients (for comparison of truly invasive and colonizing Candida spp...
2016: PloS One
E Román, I Correia, A Salazin, C Fradin, T Jouault, D Poulain, F-T Liu, J Pla
The Cek1 MAP kinase (MAPK) mediates vegetative growth and cell wall biogenesis in the fungal pathogen Candida albicans. Alterations in the fungal cell wall caused by a defective Cek1‑mediated signaling pathway leads to increased β‑1,3‑glucan exposure influencing dectin‑1 fungal recognition by immune cells. We show here that cek1 cells also display an increased exposure of α‑1,2 and β‑1,2‑mannosides (α‑M and β‑M), a phenotype shared by strains defective in the activating MAPKK Hst7, suggesting a general defect in cell wall assembly...
July 3, 2016: Virulence
Nidia Alvarez-Rueda, Audrey Fleury, Cédric Logé, Fabrice Pagniez, Estelle Robert, Florent Morio, Patrice Le Pape
Resistance to fluconazole antifungal is an ongoing impediment to a successful treatment of Candida albicans infections. One of the most prevalent mechanisms leading to azole resistance is genetic alterations of the 14α-demethylase, the target of azole antifungals, through point mutations. Site-directed mutagenesis and molecular modeling of 14α-demethylase rationalize biological data about the role of protein substitutions in the azole treatment failure. In this work, we investigated the role of N136Y substitution by site-directed mutagenesis into Pichia pastoris guided by structural analysis...
October 1, 2016: Medical Mycology: Official Publication of the International Society for Human and Animal Mycology
María T Andrés, Maikel Acosta-Zaldívar, José F Fierro
Human lactoferrin (hLf) is a protein of the innate immune system which induces an apoptotic-like process in yeast. Determination of the susceptibility to lactoferrin of several yeast species under different metabolic conditions, respiratory activity, cytoplasmic ATP levels, and external medium acidification mediated by glucose assays suggested plasma membrane Pma1p (P3A-type ATPase) as the hLf molecular target. The inhibition of plasma membrane ATPase activity by hLf and the identification of Pma1p as the hLf-binding membrane protein confirmed the previous physiological evidence...
July 2016: Antimicrobial Agents and Chemotherapy
Zdena Palková, Libuše Váchová
Yeasts, historically considered to be single-cell organisms, are able to activate different differentiation processes. Individual yeast cells can change their life-styles by processes of phenotypic switching such as the switch from yeast-shaped cells to filamentous cells (pseudohyphae or true hyphae) and the transition among opaque, white and gray cell-types. Yeasts can also create organized multicellular structures such as colonies and biofilms, and the latter are often observed as contaminants on surfaces in industry and medical care and are formed during infections of the human body...
September 2016: Seminars in Cell & Developmental Biology
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