Background. Azole resistance is an emerging problem in Aspergillus fumigatus and complicates the management of patients with Aspergillus-related diseases. Selection azole may occur through exposure to fungicides environment. In Netherlands a surveillance network was used investigate epidemiology selection A. fumigatus. Methods. Clinical isolates were screened for 8 university hospitals using agar dilution plates. Patient information collected online questionnaire azole-resistant analyzed...

RIG-I and MDA5 are cytosolic RNA sensors that play a critical role in innate antiviral responses. Major advances have been made identifying ligands, but our knowledge of the ligands for remains restricted to data from transfection experiments mostly using poly(I:C), synthetic dsRNA mimic. Here, we dissected IFN-α/β-stimulatory activity different viral species produced during picornavirus infection, both by infected cells which specific steps replication were inhibited. Our results show...

The aim of this study was to determine the applicability polymerase chain reaction (PCR) for routine diagnostic use and detection persistent enteroviral infections. To end, general primers were selected in highly conserved part 5'-noncoding region genome. They tested on 66 different enterovirus serotypes. A specific fragment amplified from 60 An amplification product not observed coxsackievirus types A11, A17, A24 echovirus 16, 22, 23. Enteroviral RNA detected by PCR routinely collected...

The family Picornaviridae contains well-known human pathogens (e.g., poliovirus, coxsackievirus, rhinovirus, and parechovirus). In addition, this a number of viruses that infect animals, including members the genus Cardiovirus such as Encephalomyocarditis virus (EMCV) Theiler's murine encephalomyelits (TMEV). latter are important cause myocarditis, type 1 diabetes chronic inflammation in brains, mimicking multiple sclerosis. Recently, new picornavirus was isolated from humans, named Saffold...

This study investigated the dynamics of Aspergillus fumigatus azole-resistant phenotypes in two compost heaps with contrasting azole exposures: free and exposed. After heat shock, to which sexual but not asexual spores are highly resistant, azole-free yielded 98% (49/50) wild-type 2% (1/50) isolates, whereas azole-containing 9% (4/45) 91% (41/45) resistant isolates. From latter compost, 80% (36/45) isolates contained TR46/Y121F/T289A genotype, (1/45) harbored TR46/Y121F/M172I/T289A/G448S had...

Viral infection of mammalian cells triggers the synthesis and secretion type I interferons (i.e. IFN-α/β), which induce transcription genes that cause to adopt an antiviral state. Many viruses have adapted mechanisms evade IFN-α/β-mediated responses. The leader protein mengovirus, a picornavirus, has been implicated as IFN-α/β antagonist. Here, we show inhibits both presence zinc finger motif in its N-terminus phosphorylation threonine-47 are required for this function. Transcription relies...

To determine the MIC distributions of itraconazole, voriconazole and posaconazole non-azole drugs for wild-type cyp51A, as well TR34/L98H TR46/Y121F/T289A cyp51A mutants Aspergillus fumigatus. We retrieved sequence data 952 clinical A. fumigatus strains isolated in or referred to our reference laboratory, during January 2010 December 2013 period. All MICs were determined using EUCAST methodology interpreted breakpoints. Three-hundred sixty-four (38%) resistant azoles. Of these, 225 contained...

ABSTRACT In a retrospective multicenter study, 102 formalin-fixed paraffin-embedded (FFPE) tissue specimens with histopathology results were tested. Two 4- to 5-μm FFPE sections from each specimen digested proteinase K, followed by automated nucleic acid extraction. Multiple real-time quantitative PCR (qPCR) assays targeting the internal transcribed spacer 2 (ITS2) region of ribosomal DNA, using fluorescently labeled primers, was performed identify clinically important genera and species...

In order to survive, Aspergillus fumigatus must adapt specific niche environments. Adaptation the human host includes modifications facilitating persistent colonisation and development of azole resistance. The aim this study is advance understanding genetic physiological adaptation A. in patients during infection treatment. Thirteen strains were isolated from a single chronic granulomatous disease patient suffering recurrent invasive aspergillosis over period 2 years. All had identical...

Olorofim (F901318) is a new antifungal currently under clinical development that shows both in vitro and vivo activity against number of filamentous fungi including Aspergillus fumigatus. In this study, we screened A. fumigatus isolates for intrinsic olorofim-resistant evaluated the ability to acquire an phenotype. No resistance was found 975 isolates. However, with increased olorofim MICs (> 8 mg/L) could be selected using high conidia exposure laboratory conditions. Assessment frequency...

A resistant and hypervirulent dermatophyte from India has been described as a taxonomic novelty, Trichophyton indotineae, species of the mentagrophytes complex. Rapid detection correct identification closely similar dermatophytes with different predilections are essential for efficient clinical management. We evaluated efficacy rapid diagnostic methods environmental strains in T. The included Real-time-PCR, DermaGenius, LAMP, MALDI-ToF MS, using rDNA ITS sequences standard. results show that...

ICU-acquired muscle weakness commonly occurs in patients with septic shock and is associated poor outcome. Although atrophy known to be involved, it unclear whether ligands plasma from these are responsible for initiating degradation of proteins. The aim the present study was investigate if induces skeletal examine time course plasma-induced during ICU stay.Plasma derived within 24 hours after hospital admission (n = 21) healthy controls 12). From nine additionally at two, five seven days...

Enterovirus protein 2B has been shown to increase plasma membrane permeability. We have identified a conserved putative amphipathic alpha-helix with narrow hydrophilic face and an arrangement of cationic residues that is typical for the so-called lytic polypeptides. To examine functional structural roles this alpha-helix, we constructed nine coxsackie B3 virus mutants by site-directed mutagenesis infectious cDNA clone. Six contained substitutions charged in alpha-helix. Three insertions...

Coxsackie B virus protein 2B contains near its C terminus a hydrophobic domain with an amino acid composition that is characteristic for transmembrane regions. A molecular genetic approach was followed to define the role of this in reproduction and study structural requirements domain. Nine substitution mutations were introduced infectious cDNA clone coxsackie B3 virus. The effects studied vivo by transfection Buffalo green monkey cells copy RNA transcripts. results reported here suggest...

Antifungal susceptibility testing is an essential tool for guiding therapy, although EUCAST and CLSI reference methods are often available only in specialized centers. We studied the performance of agar-based screening method detection azole resistance Aspergillus fumigatus cultures. The VIPcheck consists four wells containing voriconazole, itraconazole, posaconazole, or a growth control. Ninety-six A. isolates were used. Thirty-three harbored known mechanism: TR34/L98H (11 isolates),...

In the last few years, increasing numbers of viruses infecting fungi have been identified. this study, we used an in silico approach for analysis deep RNA sequencing data order to discover and characterize putative genomic ssRNA or dsRNA mycovirus sequences Aspergillus fumigatus. reads A. fumigatus strains were mapped against Af293 reference genome. Unmapped collected de novo assembly. Contigs analyzed by Blastx comparison with a protein database. Assembled viral genomes as template...

In our studies on the biological function of mengovirus leader protein, we identified a casein kinase II (CK-2) phosphorylation site in protein. Here report that protein can be phosphorylated by CK-2 vitro. Expression recombinant which consensus sequence around threonine 47 was disturbed resulted mutant could no longer phosphorylated. The modified site-directed mutagenesis and subsequently introduced into cDNA clone to investigate effect virus replication host cell response. Modifications...

Aspergillus fumigatus is a saprophytic fungus that extensively produces conidia. These microscopic asexually reproductive structures are small enough to reach the lungs. Germination of conidia followed by hyphal growth inside human lungs key step in establishment infection immunocompromised patients. RNA-Seq was used analyze transcriptome dormant and germinating A. Construction gene co-expression network revealed four clusters (modules) correlated with phase (dormant, isotropic growth,...