The present study examines the trend in distribution of Candida species and their antifungal resistance patterns in hospitals across Haryana, a North Indian state with poorly addressed epidemiology of fungal infections. In our collection of 228 Candida isolates, Candida albicans dominated in both high vaginal swab (HVS) and urine samples while Candida glabrata and Candida tropicalis were the second-highest non-albicans Candida species (NAC), respectively. Of note, in blood samples, C. tropicalis and C. albicans were present in equal numbers. All 228 isolates were subjected to antifungal susceptibility tests, whereby 51% of C. albicans recovered from HVS samples displayed fluconazole resistance. To understand its mechanistic basis, expression profiling of efflux pump genes CDR1, CDR2, MDR1 and azole drug target, ERG11 was performed in 20 randomly selected resistant isolates, wherein many isolates elicited higher expression. Further, ERG11 gene sequencing suggested that most of the isolates harbored mutations, which are not reported with azole resistance. However, one isolate, RPCA9 (MIC 64 μg/mL) harbored triple mutation (Y132C, F145L, A114V), wherein Y132 and F145 sites were previously implicated in azole resistance. Interestingly, one isolate, (RPCA61) having MIC > 128 μg/mL harbored a novel mutation, G129R. Of note, HVS isolates RPCA 21, RPCA 22, and RPCA 44 (MICs 64 to > 128 μg/mL) did not show any change in alteration in ERG11 or overexpression of efflux pump genes. Together, this study presents a first report of Candida infections in selected hospitals of Haryana State.
- MeSH
- antifungální látky farmakologie MeSH
- azoly farmakologie MeSH
- Candida albicans účinky léků genetika izolace a purifikace MeSH
- Candida klasifikace účinky léků genetika izolace a purifikace MeSH
- fungální léková rezistence genetika MeSH
- fungální proteiny genetika MeSH
- geny MDR genetika MeSH
- kandidóza epidemiologie mikrobiologie MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- mutace MeSH
- nemocnice MeSH
- regulace genové exprese u hub MeSH
- retrospektivní studie MeSH
- systém (enzymů) cytochromů P-450 genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Indie MeSH
Human pathogens belonging to the Nakaseomyces clade include Candida glabrata sensu stricto, Candida nivariensis and Candida bracarensis. Their highly similar phenotypic characteristics often lead to misidentification by conventional laboratory methods. Therefore, limited information on the true epidemiology of the Candida glabrata species complex is available. Due to life-threatening infections caused by these species, it is crucial to supplement this knowledge. The aim of the study was to estimate the prevalence of C. bracarensis and C. nivariensis in a culture collection of C. glabrata complex isolates. The study covered 353 isolates identified by biochemical methods as C. glabrata, collected from paediatric and adult patients hospitalised at four medical centres in Southern Poland. The multiplex PCR was used to identify the strains. Further species confirmation was performed via sequencing and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) analysis. One isolate was recognised as C. bracarensis (0.28%). To our knowledge, it is the first isolate in Poland. C. glabrata sensu stricto species has been confirmed for all the remaining isolates. No C. nivariensis was found. Our study has shown that the prevalence of C. nivariensis and C. bracarensis strains is infrequent. However, it should be emphasised that the incidence of these strains may differ locally and depend on environmental factors and the population.
- MeSH
- antibakteriální látky farmakologie MeSH
- banky biologického materiálu MeSH
- Candida glabrata genetika MeSH
- Candida účinky léků genetika izolace a purifikace MeSH
- dospělí MeSH
- hospitalizace MeSH
- kandidóza mikrobiologie MeSH
- kultivační média MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- multiplexová polymerázová řetězová reakce MeSH
- předškolní dítě MeSH
- prevalence MeSH
- sekvenční analýza DNA MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- předškolní dítě MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Polsko MeSH
Candida krusei is a pathogenic yeast species that is phylogenetically outside both of the well-studied yeast groups, whole genome duplication and CUG. Like all other yeast species, it needs to accumulate high amounts of potassium cations, which are needed for proliferation and many other cell functions. A search in the sequenced genomes of nine C. krusei strains revealed the existence of two highly conserved genes encoding putative potassium uptake systems. Both of them belong to the TRK family, whose members have been found in all the sequenced genomes of species from the Saccharomycetales subclade. Analysis and comparison of the two C. krusei Trk sequences revealed all the typical features of yeast Trk proteins but also an unusual extension of the CkTrk2 hydrophilic N-terminus. The expression of both putative CkTRK genes in Saccharomyces cerevisiae lacking its own potassium importers showed that only CkTrk1 is able to complement the absence of S. cerevisiae's own transporters and provide cells with a sufficient amount of potassium. Interestingly, a portion of the CkTrk1 molecules were localized to the vacuolar membrane. The presence of CkTrk2 had no evident phenotype, due to the fact that this protein was not correctly targeted to the S. cerevisiae plasma membrane. Thus, CkTrk2 is the first studied yeast Trk protein to date that was not properly recognized and targeted to the plasma membrane upon heterologous expression in S. cerevisiae.
- MeSH
- Candida klasifikace genetika růst a vývoj metabolismus MeSH
- draslík metabolismus MeSH
- fungální proteiny genetika metabolismus MeSH
- fylogeneze MeSH
- genetická variace MeSH
- genom fungální genetika MeSH
- iontový transport MeSH
- proteiny přenášející kationty genetika metabolismus MeSH
- rekombinantní proteiny genetika metabolismus MeSH
- Saccharomyces cerevisiae klasifikace genetika růst a vývoj metabolismus MeSH
- Saccharomycetales klasifikace genetika MeSH
- testy genetické komplementace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Candida dubliniensis was reported as a new species in 1995. This species is often misidentified as Candida albicans. The aims of this work were to determine the occurrence of C. dubliniensis in various clinical materials, to evaluate several ways to identify it and to examine the genetic variability of isolates. Among 7706 isolates originally identified as C. albicans, 237 were identified as C. dubliniensis (3.1%). Most of the C. dubliniensis isolates were obtained from the upper and lower respiratory tract (61.4 and 22.9%). Five phenotypic methods including latex agglutination were used (cultivation on CHROMagar Candida, on Staib agar, at 42 °C and in medium with 6.5% NaCl), but only cultivation on the medium with an increased concentration of NaCl and latex agglutination gave reliable results. Species-specific polymerase chain reaction was used as the confirmation method. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry provided less reliable results. In fact, 78.9% of C. dubliniensis isolates had scores above 1.7. However, the rest of them (21.1%) were also identified as C. dubliniensis even when the scores were lower than 1.7. Divergences among C. dubliniensis strains were evaluated by means of pulsed-field gel electrophoresis. Eighty-six selected C. dubliniensis isolates showed a 69.6% level of similarity. The results of this study expand the knowledge of the incidence, means of identification and genotypic divergence of C. dubliniensis isolates.
- MeSH
- Candida klasifikace genetika izolace a purifikace fyziologie MeSH
- chlorid sodný metabolismus MeSH
- genetická variace MeSH
- genotyp MeSH
- kandidóza mikrobiologie MeSH
- latex fixační testy MeSH
- lidé MeSH
- mikrobiologické techniky metody MeSH
- polymerázová řetězová reakce MeSH
- pulzní gelová elektroforéza MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- teplota MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Yeasts frequently colonize non-sterile sites in the body. The aim of the study was to determine distribution in clinical samples and antifungal susceptibility to five antifungals. From January 2013 through June 2015, 800 isolates were obtained from intensive care unit patients. Candida albicans (58.9%), Candida glabrata (20.4%), Candida krusei (8.6%), and Candida parapsilosis (3.6%) were the leading species. Majority of the C. albicans isolates were susceptible to the fluconazole. Elevated voriconazole minimal inhibitory concentrations (MICs) were observed in isolates exhibiting high fluconazole MICs, most frequently in C. glabrata. Isolates with echinocandins MICs suggesting reduced susceptibility were only sporadic cases with the exception of Trichosporon spp. The amphotericin B MICs were slightly higher for some C. krusei.
- MeSH
- amfotericin B farmakologie MeSH
- antifungální látky farmakologie MeSH
- Candida klasifikace účinky léků genetika izolace a purifikace MeSH
- flukonazol farmakologie MeSH
- fungální léková rezistence MeSH
- jednotky intenzivní péče statistika a číselné údaje MeSH
- kandidóza mikrobiologie MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- vorikonazol farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Patogenní kvasinky rodu Candida představují nejrozšířenější příčinu mykotických onemocnění. Tyto druhy jsou úspěšnými patogeny díky tomu, že jsou schopné se rozmnožovat v nejrůznějších místech jako je například kůže, gastrointestinální trakt, krev nebo vaginální sliz- nice a také díky schopnosti adheze na abiotické povrchy. Jednou z klíčových strategií přežití těchto patogenů je jejich schopnost poliferovat v různých koncentracích oxidu uhličitého (CO2). K zvládnutí takového rozdílu používají kvasinky rodu Candida karbanické anhydrasy (CA) kódované homologem genu NCE103, které katalyzují reverzibilní hydrataci CO2 na bikarbonát. CA jsou postradatelné během infekce krevního řečiště, ale jsou nezbytné pro přežití kvasinek na kůži nebo na abiotických površích. Kvasinkové CA jsou strukturně odlišné od lidských a představují tak vhodný cíl pro vývoj profylaktických látek.
Pathogenic yeasts of the genus Candida represent the most prevalent cause of mycotic diseases worldwide. These species are successful pathogens due to their ability to proliferate under a wide variety of conditions, colonizing host niches as diverse as skin, blood, or vagina and also to adhere to abiotic surfaces. One of the key survival strategies of fungal pathogens is the ability to proliferate in different carbon dioxide (CO2) concentrations. To cope with such difference, Candida possesses carbonic anhydrase (CA), encoded by NCE103 gene, which catalyzes reversible hydratation of CO2 into bicarbonate. This enzyme is dispensable during the bloodstream infection, but it is essential for survival of the fungus on skin or abiotic surfaces. Fungal CAs are structurally unrelated to human CAs, which makes them an ideal target for prophylactic intervention.
- Klíčová slova
- NCE103,
- MeSH
- antifungální látky farmakologie klasifikace MeSH
- Candida enzymologie genetika patogenita MeSH
- fungální léková rezistence účinky léků MeSH
- houby enzymologie MeSH
- kandidóza invazivní patologie terapie MeSH
- kandidóza etiologie farmakoterapie klasifikace patologie MeSH
- karboanhydrasy * genetika chemie klasifikace účinky léků MeSH
- kvasinky * enzymologie klasifikace patogenita MeSH
- oxid uhličitý MeSH
- Publikační typ
- práce podpořená grantem MeSH
To study the mechanisms involved in the maintenance of a linear mitochondrial genome we investigated the biochemical properties of the recombination protein Mgm101 from Candida parapsilosis. We show that CpMgm101 complements defects associated with the Saccharomyces cerevisiae mgm101-1(ts) mutation and that it is present in both the nucleus and mitochondrial nucleoids of C. parapsilosis. Unlike its S. cerevisiae counterpart, CpMgm101 is associated with the entire nucleoid population and is able to bind to a broad range of DNA substrates in a non-sequence specific manner. CpMgm101 is also able to catalyze strand annealing and D-loop formation. CpMgm101 forms a roughly C-shaped trimer in solution according to SAXS. Electron microscopy of a complex of CpMgm101 with a model mitochondrial telomere revealed homogeneous, ring-shaped structures at the telomeric single-stranded overhangs. The DNA-binding properties of CpMgm101, together with its DNA recombination properties, suggest that it can play a number of possible roles in the replication of the mitochondrial genome and the maintenance of its telomeres.
- MeSH
- buněčné jádro genetika metabolismus MeSH
- Candida genetika metabolismus MeSH
- DNA fungální genetika metabolismus MeSH
- DNA vazebné proteiny genetika metabolismus MeSH
- Escherichia coli genetika metabolismus MeSH
- exprese genu MeSH
- genom fungální * MeSH
- genom mitochondriální * MeSH
- homeostáza telomer MeSH
- klonování DNA MeSH
- mitochondriální proteiny genetika metabolismus MeSH
- mitochondrie genetika metabolismus MeSH
- multimerizace proteinu MeSH
- mutace MeSH
- regulace genové exprese u hub * MeSH
- rekombinace genetická MeSH
- rekombinantní proteiny genetika metabolismus MeSH
- Saccharomyces cerevisiae - proteiny genetika metabolismus MeSH
- Saccharomyces cerevisiae genetika metabolismus MeSH
- telomery chemie metabolismus MeSH
- testy genetické komplementace MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Yeast mtDNA is compacted into nucleoprotein structures called mitochondrial nucleoids (mt-nucleoids). The principal mediators of nucleoid formation are mitochondrial high-mobility group (HMG)-box containing (mtHMG) proteins. Although these proteins are some of the fastest evolving components of mt-nucleoids, it is not known whether the divergence of mtHMG proteins on the level of their amino acid sequences is accompanied by diversification of their biochemical properties. In the present study we performed a comparative biochemical analysis of yeast mtHMG proteins from Saccharomyces cerevisiae (ScAbf2p), Yarrowia lipolytica (YlMhb1p) and Candida parapsilosis (CpGcf1p). We found that all three proteins exhibit relatively weak binding to intact dsDNA. In fact, ScAbf2p and YlMhb1p bind quantitatively to this substrate only at very high protein to DNA ratios and CpGcf1p shows only negligible binding to dsDNA. In contrast, the proteins exhibit much higher preference for recombination intermediates such as Holliday junctions (HJ) and replication forks (RF). Therefore, we hypothesize that the roles of the yeast mtHMG proteins in maintenance and compaction of mtDNA in vivo are in large part mediated by their binding to recombination/replication intermediates. We also speculate that the distinct biochemical properties of CpGcf1p may represent one of the prerequisites for frequent evolutionary tinkering with the form of the mitochondrial genome in the CTG-clade of hemiascomycetous yeast species.
- MeSH
- Candida genetika metabolismus MeSH
- mitochondriální proteiny genetika metabolismus MeSH
- molekulární evoluce * MeSH
- proteiny s vysokou pohyblivostí genetika metabolismus MeSH
- Saccharomyces cerevisiae - proteiny genetika metabolismus MeSH
- Saccharomyces cerevisiae genetika metabolismus MeSH
- Yarrowia genetika metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Candida parapsilosis produces secreted aspartic proteinases (Saps), which contribute to the virulence of this opportunistic pathogen. Gene family containing as many as 14 sequences potentially encoding secreted aspartic proteinases was identified in C. parapsilosis genome. Of them, SAPP1 and SAPP2 genes have been extensively characterized, but only now do we report that two SAPP2 homologs sharing 91.5 % identity occur in C. parapsilosis genome. Existence of SAPP2 homologs points to unexpected complexity of the SAPP gene family.
- MeSH
- aspartátové proteasy chemie genetika metabolismus MeSH
- Candida chemie enzymologie genetika MeSH
- fungální proteiny chemie genetika metabolismus MeSH
- genom fungální * MeSH
- molekulární sekvence - údaje MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The virulence of the Candida pathogens is enhanced by the production of secreted aspartic proteases, which therefore represent possible targets for drug design. Here, the crystal structure of the secreted aspartic protease Sapp2p from Candida parapsilosis was determined. Sapp2p was isolated from its natural source and crystallized in complex with pepstatin A, a classical aspartic protease inhibitor. The atomic resolution of 0.83 Å allowed the protonation states of the active-site residues to be inferred. A detailed comparison of the structure of Sapp2p with the structure of Sapp1p, the most abundant C. parapsilosis secreted aspartic protease, was performed. The analysis, which included advanced quantum-chemical interaction-energy calculations, uncovered molecular details that allowed the experimentally observed equipotent inhibition of both isoenzymes by pepstatin A to be rationalized.
- MeSH
- aspartátové proteasy chemie genetika izolace a purifikace metabolismus MeSH
- Candida chemie enzymologie genetika MeSH
- exprese genu MeSH
- fungální proteiny chemie genetika izolace a purifikace metabolismus MeSH
- inhibitory proteas chemie MeSH
- izoenzymy chemie genetika izolace a purifikace metabolismus MeSH
- katalytická doména MeSH
- kinetika MeSH
- krystalografie rentgenová MeSH
- kvantová teorie MeSH
- molekulární modely MeSH
- molekulární sekvence - údaje MeSH
- pepstatiny chemie MeSH
- sekundární struktura proteinů MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- strukturní homologie proteinů MeSH
- substrátová specifita MeSH
- termodynamika MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
