Fungal Genet Biol 48:15–22PubMed Bungihan ME, Tan MA, Kitajima M,

Fungal Genet Biol 48:15–22PubMed Bungihan ME, Tan MA, Kitajima M, Kogure N, Franzblau SG, dela Cruz TEE, Takayama H, Nonato MG (2011) Bioactive metabolites of Diaporthe sp. P133, an endophytic fungus isolated from Pandanus amaryllifolius. J Nat Med 65:606–609PubMed Burns E, Ifrach I, Carmeli S, Pawlik JR, Ilan M (2003) Comparison of antipredatory sedimentary lipids? Org Geochem 30:1–14 Cheng MJ, Wua MD, Yuan GF, Chen YL, Su YS, Hsieh MT, Chen IS (2012) Secondary metabolites and cytotoxic activities from

the endophytic fungus Annulohypoxylon squamulosum. Phytochem Lett 5:219–223 Cichewicz RH (2010) Epigenome manipulation as a pathway AZD6244 manufacturer to new natural product scaffolds and their congeners. Nat Prod Rep 27:11–22PubMed Cichewicz RH, Clifford LJ, Lassen PR, Cao X, Freedman TB, Nafie LA,

JNJ-64619178 ic50 Deschamps JD, Kenyon VA, Flanary JR, Holman TR, Crews P (2005) Stereochemical determination and bioactivity assessment of (S)-(+)-curcuphenol dimers isolated from the marine sponge Didiscus aceratus and synthesized through laccase biocatalysis. Bioorg Med Chem 13:5600–5612PubMed Cohen E, Koch L, Thu KM, Rahamim Y, Aluma Y, Ilan M, Yarden O, Carmeli S (2011) Novel terpenoids of the fungus Aspergillus insuetus isolated from the Mediterranean sponge Psammocinia sp. collected along the coast of Israel. Bioorg Med Chem 19:6587–6593PubMed Córdoba-Pedregosa MC, Villalba JM, Córdoba F, González-Reyes JA (2005) Changes in intracellular and apoplastic peroxidise activity, ascorbate redox status and root elongation induced by EPZ015938 datasheet enhanced ascorbate content in Allium cepa L. J Exp Bot 56:685–694 Criddle RS, Hansen LD, Breidenbach RW, Ward MR, Huffaker RC (1989) Effects of NaCl on metabolic heat evolution rates by barley roots. Plant Physiol 90:53–58PubMed Debbab

A, Aly AH, Edrada-Ebel RA, Wray V, Müller WEG, Totzke F, Zirrgiebel U, Schächtele C, Kubbutat MHG, Lin WH, Mosaddak M, Hakiki A, Proksch P, Ebel R (2009) Bioactive metabolites from endophytic fungus Stemphylium globuliferum isolated from Mentha pulegium. J Nat Prod Vitamin B12 72:626–631PubMed Debbab A, Aly AH, Lin WH, Proksch P (2010) Bioactive compounds from marine bacteria and fungi. Microbiol Biotechnol 3:544–563 Debbab A, Aly AH, Proksch P (2011) Bioactive secondary metabolites from terrestrial endophytes and associated marine derived fungi. Fungal Divers 49:1–12 Debbab A, Aly AH, Edrada-Ebel R, Wray V, Pretsch A, Pescitelli G, Kurtan T, Proksch P (2012) New anthracene derivatives—structure elucidation and antimicrobial activity. Eur J Org Chem 1351–1359. Ding B, Yin Y, Zhang F, Li Z (2011) Recovery and phylogenetic diversity of culturable fungi associated with marine sponges Clathrina luteoculcitella and Holoxea sp. in the South China Sea. Mar Biotechnol 13:713–721PubMed Ding G, Hl W, Chen L, Chen AJ, Lan J, Chen XD, Zhang HW, Chen H, Liu XZ, Zou ZM (2012) Cytochalasans with different amino-acid origin from the plant endophytic fungus Trichoderma gamsii.

On the other hand, deletion of specific CW proteins sensitize yea

On the other hand, deletion of specific CW proteins sensitize yeast to the antibacterial lantibiotic nisin [66]. Further, PAF26 induced severe mycelial growth and cell-shape defects to the fungus P. digitatum [46], changes that are typical for compounds affecting the cell wall. Our assays showed only a limited number of gene deletions related to CW that have an effect on sensitivity to PAF26 or melittin. Even in these examples, the magnitude of the phenotype of the mutants (i.e., changes in sensitivity)

is modest compared to that of mutants related to ribosome biogenesis, arginine metabolism, sphingolipid or HSP related genes (compare Figures 4 and 5). This LY3023414 mouse holds even for genes such as the above mentioned SSD1, which mediates deposition of other CW proteins in S. cerevisiae [56]. The corresponding deletion strain has a damaged CW as confirmed by hyper-sensitivity to SDS

or CFW, but comparatively only a minor increase in susceptibility to AMP as demonstrated in two genetic backgrounds C646 price (BY4741 and RAY3A, see also Additional File 6). A similar phenotype was observed in other strains such as Δecm33. Microscopy and flow cytometry studies in Δssd1 or Δecm33 showed a correlation between a higher sensitivity and an increase of PAF26 uptake of cells (Figure 7), demonstrating that CW components modulate the interaction with peptides. Function redundancy might explain the lack of a dramatic change in the susceptibility in mutants related to CW. Therefore multiple deletions

would be expected to have a higher impact and are being studied in our laboratory. However, our current data do not support this view either, as illustrated with the triple deletion of PIR genes in the RAY3A background (Additional File 6). Even in the case of gene deletions from MAPK Thiazovivin cost signalling cascades involved in CW construction and response to stress [51], we did not find major differences in sensitivity Adenosine triphosphate to peptides under our assay conditions (Additional File 7). Representative examples are STE2 that was highly repressed by both peptides, or SLT2, PBS2 and HOG1, whose deletants are hypersensitive to CW interfering compounds. This result contrasts with previous data in which mutations in the HOG osmoregulatory pathway in the case of the peptide histatin [31] or the RHO1-SLT2 CW growth pathway in a plant defensin Pn-AMP1 [67] result in hypersensitivity. Other CW-related gene deletions did not show significant differences in susceptibility to peptides and even in a limited number of examples (as Δsed1) a slight higher resistance was observed. It has been described that specific gene deletions result in counteracting mechanisms to reinforce CW by enhancing levels of specific CW constituents [64].

3 eV) which could reduce the energy barrier for carrier transport

3 eV) which could reduce the energy barrier for carrier transport and also effectively avoid parasitic absorption. However,

doped Si-NCs embedded in a SiN x matrix (Si-NCs/SiN x ) have not received much attention. In this work, we present initial fabrication and characterization results of Si heterojunction solar cells find more using P-doped Si-NCs/SiN x films as emitters. The P-doped Si-NCs/SiN x films were prepared by electron cyclotron resonance chemical vapor deposition (ECRCVD) followed by high-temperature annealing, and the influence of the chemical composition (N/Si ratio) on their physical properties was investigated. The photovoltaic properties of the fabricated heterojunction devices were also examined as a function of the N/Si composition ratio in the P-doped Si-NCs/SiN x films. Methods Fifty-nanometer-thick, homogeneous Si-rich silicon nitride (SRN) films containing phosphorus were deposited by a homemade ECRCVD system

on single-side polished p-type (100) single crystalline Si (sc-Si) substrates with a thickness of 675 μm and a AZD1480 cost resistivity in the range of 5 to 20 Ω cm. Before placing into the deposition Nutlin-3a concentration chamber, Si substrates were cleaned with acetone and rinsed in deionized water followed by removal of native oxide on Si wafers using a diluted HF dip (5%). The mixed SiH4, N2, Ar, and PH3 gases were then introduced into the deposition chamber at 10 mTorr for film growth. The applied microwave power and the Apoptosis inhibitor substrate temperature were kept on

700 W and 200°C, respectively. In order to study the influence of the Si/N ratio on film properties, both SiH4 and PH3 flow rates were kept constant during film growth, while the gas mix ratio (R c) defined as N2/SiH4 was varied in the range 0.73 ≤ R c ≤ 0.83. The formation of Si-NCs in as-deposited SRN films was carried out by post-growth annealing in a quartz tube furnace at 950°C in N2 ambient. Samples with a 1 cm × 1 cm area were used for subsequent fabrication of heterojunction solar cells. Aluminum films deposited by electron gun evaporation were used as contact electrode layers in the solar cells. The front contact on top of the Si-NCs/SiN x film was defined by a shadow mask during Al deposition, while the rear contact covered the full back area of the cell. After metallization, the samples were heated at 500°C for 3 min to improve the electrical properties of the contacts. For the characterization, the bonding configurations of the Si-NCs/SiN x films were identified by X-ray photoelectron spectroscopy (XPS). Micro-Raman spectroscopy and transmission electron microscopy (TEM) were used to investigate the crystallization behavior in SRN films after post-growth annealing. Fused quartz wafers were used as substrates for Raman studies to avoid the signal contribution from Si substrates during Raman measurements. X-ray diffraction (XRD) was used to evaluate the Si-NC size of annealed samples.

Fluorescent images were analyzed by the GenePixPro software (v 6

Fluorescent images were analyzed by the GenePixPro software (v.6.0) and Acuity (v.4.0) (Axon Instruments). The intensity of fluorescence PCI32765 of each spot was measured and the mean of 4 replicate spots per probe was calculated.

Local background fluorescence was also measured and subtracted from the mean fluorescence. Spots displaying fluorescence greater than mean fluorescence of all spots on the array plus two times standard deviation (SD) were considered as positive. The hybridization was considered successful if spiked and control spots produced positive signals. Presence of more than 5 positive spots from same species was interpreted as positivity of the sample for this pathogen species. The fidelity limit of LSplex was defined as minimal amount of DNA necessary to obtain the hybridization pattern with >95% correspondence to one from the 2 μg genomic DNA. Results We have recently established a prototype medium-density gene-segment DNA microarray for the detection and genetic profiling of pathogens causing bloodstream Elacridar infections [2]. The limit of detection of such medium-density gene-segment DNA microarrays was previously identified and ranged between 10 and 100 ng of DNA [2]. This microarray has been 3-deazaneplanocin A cell line extended for the present study to represent specific gene fragments of more than 20 of the most prominent causative agents of sepsis [15]. As expected the sensitivity

of detection was not influenced by the extension of the microarray. This was confirmed experimentally by hybridizing decreasing amounts of bacterial genomic DNA (Additional file 2). At the nanogram level a striking reduction Cobimetinib chemical structure in the detection power was observed and the number of detected genes was gradually reduced. In order to improve the sensitivity of detection we focused on the development of an amplification protocol by multiplex PCR. Large scale multiplex PCR with 800-primer pairs (LSplex)

The amplification of unidentified pathogen DNA requires that all necessary primer pairs are present in the amplification mix. We have initially addressed the question whether it is possible to amplify genomic DNA of several bacterial species by a PCR containing 800 primer pairs (Additional file 1). However, the complexity of the primer mix did not allow the amplification of any genomic DNA at a final primer concentration of 0.2 μM (data not shown). Nevertheless, reducing the primer concentration in the amplification reaction to 0.02 μM permitted amplification from 100 ng of some DNA templates, although the amplification of most DNA templates was very weak (Fig 1A). It was not possible to further decrease the final concentration of individual primers without a negative effect on the amplification yield (not shown). Furthermore, DNA templates from Gram-negative bacteria could not be amplified using Taq DNA polymerase at any primer concentration (not shown).

Two sets of study data will be evaluated: the primary

Two sets of study data will be evaluated: the primary AZD6244 concentration objective will be

evaluated in the full analysis set (FAS). The FAS is defined as the set of data generated from the included patients who received at least the safety dose. The secondary objectives will be evaluated in both FAS and per-protocol set (PPS). The PPS is defined as the set of data generated from the included patients who complied with the protocol. Monitoring The IDMC will perform a safety review after each series of treatments of three consecutive patients. The IDMC members have no conflict of interest with the sponsor because they are not involved in the study, nor are they receiving funds. The IDMC will work according to standard operating procedures and will receive reports on a regular JNJ-64619178 in vivo basis on all toxicity CTCAE ≥ grade 3 reported for this trial. Recruitment will not be interrupted unless otherwise requested by the chairman of the IDMC. The responsibilities of the IDMC include:

minimize the exposure of patients to an unsafe therapy or dose make recommendations for changes in study processes where appropriate endorse continuation of the study inform the institutional IEC in the case of toxicity CTCAE ≥ grade 3 and/or when the well-being of the subjects is jeopardized Ethical considerations The study will be conducted according to the principles of the Declaration of Helsinki (version 9.10.2004) and in accordance with the Medical Research Involving Human Patients Act (WMO), the requirements of International Conference on Harmonization Bumetanide – Good Clinical Practice. The study protocol has been approved by the IEC and by the institutional Radiation Protection Committee. Discussion The HEPAR trial is a phase I study to evaluate the safety and toxicity profile of 166Ho radioembolization. Secondary endpoints are tumour response, biodistribution assessment, performance status,

quality of life and comparison of the biodistributions of the 99mTc-MAA scout dose and the 166Ho-PLLA-MS safety dose. With regard to the method of administration, viz. through a catheter placed in the hepatic artery, the in-vivo characteristics (no significant release of radionuclide), and the mechanism of action (local irradiation of the tumour), 166Ho-PLLA-MS constitute a device analogous to the 90Y microspheres, which are currently applied clinically. 166Ho-PLLA-MS only differ in the radioisotope and the device matrix that are used. In a toxicity study in pigs on 166Ho-RE, it has been demonstrated that (healthy) pigs can withstand extremely high liver absorbed doses, at least up to 160 Gy [23]. During these animal experiments, only very mild side effects were seen: slight and transitory inappetence and somnolence, which may well have been associated with the anaesthetic and Avapritinib supplier analgesic agents that had been given and not necessarily with the microsphere administration.

Resistance to these and other antibiotics in pathogenicS epiderm

Resistance to these and other antibiotics in pathogenicS. epidermidisisolates has been reported previously [10,19]. The resistance of these strains could be partly due to the increasing use of broad-spectrum antibiotics, which encourage selection of multirresistant strains [11].

Improper antibiotherapy may explain why staphylococcal mastitis frequently becomes a chronic and/or recurrent infection. In this study, the presence ofmecA gene accompanied with resistance to oxacillin (MIC > 2 μg mL-1) was observed in 62% of the strains from mastitis, but only in 33% from the healthy group. ThemecA gene was not detected in four oxacillin-resistant strains. These strains may represent cases of borderline resistance which is characterized by an oxacillin MIC at or just above the susceptibly breakpoint (4 to 8 μg mL-1). In contrast, themecA gene was detected selleck kinase inhibitor in five oxacillin-susceptible strains, a fact that has been previously described [20] and that may be due to gene deletions. Methicillin-resistantStaphylococcus

aureus(MRSA) are being reported with increasing frequency in EPZ5676 the community and they have been called community-acquired (CA)-MRSA, which are associated with skin and soft tissue infection [21] but are also frequently isolated from healthy hosts [22]. Most of themecA+strains used in this study could be ascribed to type IV SCCmec. InS. epidermidis, some studies Farnesyltransferase have reported that SCCmectype IV is generally carried by CA-MRS [23,24] but this type seems to be predominant among clinically relevantS. epidermidisisolates [9]. The fact that theccrB gene was not amplified from fourmecA+strains may be due to the presence of different alleles for this gene [25]. In the last years, a renewed medical and research interest has been focused onS. epidermidissince it has become the most important

cause of nosocomial infections [6]. The complete Lenvatinib research buy Genome analysis of some methicillin-resistantS. aureusandS. epidermidisstrains of human origin have revealed the propensity ofS. aureusto cause fulminant and sometimes life-threatening infections, as opposed to the predisposition ofS. epidermidisfor chronic and recurrent infections [26]. Identification ofS. epidermidisas etiological agents of infection is sometimes hindered by the fact that infections associated with this microorganims are characterized by subtle, non-specific clinical manifestations [5]. Precisely, these characteristics occur in most cases of lactational mastitis. Genome flexibility inS. epidermidismay contribute to the acquisition of some transferable virulence and resistant traits [6,27] and to the evolution of this species from a commensal to a pathogenic microorganism in susceptible hosts [28].

In this chapter Perrier proposes his own scenario on the origin o

In this chapter Perrier proposes his own scenario on the origin of life and shows that the phenomenon of life began

with a unique starting point on a primitive earth very different from today. He gives also some methodological keys to try to experiment in laboratory the first stages leading to life. Finally he points out some difficulties that are still topical nowadays. This paper will show what innovations had been made by Perrier in the field of the emergence of life, and why his suggestions can be regarded as very close to the first scenarios of chemical evolution. Reale, G. and Antiseri, D. (1983). Historia del Pensamiento Filosfico y Cientfico. Herder, Espaa. E-mail: [email protected]​fr Life as a Functional Concept: Functionalism as a Robust Framework for Theories PRIMA-1MET order and Definitions of Multi-realized Living Systems Olin Robus1,3, Nathan Haydon1,3, selleck kinase inhibitor Shawn McGlynn1,2, Gordon Brittan3 1NASA Astrobiology Institute; Astrobiology Biogeocatalysis Research Center; 2Department of Chemistry and Biochemistry; 3Department of History and Philosophy, Montana State University Bozeman, MT 59717 United States Past attempts defining life have been largely unsuccessful, due in part to a

flaw common to all of these attempts. Namely, these attempts are intrinsically handicapped by their formulation within a framework that implicitly assumes life is a “Natural Kind.” This characterization of life as a Natural Kind is Rucaparib ubiquitous, either implicitly or explicitly, in many definitions and theories of life. We argue that the Natural Kind paradigm falsely suggests an ontological category for living systems, and hinders investigations and exploration for non-terrestrial life. Contemporary searches for non-terrestrial living systems should rely upon a theory that can accommodate multiple

realizations of life in diverse contexts. The Natural Kind paradigm unnecessarily restricts the domain of potential realizations to an artificially small range of physical arrangements. We suggest a new conceptual framework for studying living systems, the origins of life, and the resulting theories and definitions of life, generally construed. We propose that understanding life as a functional class, rather than a Natural Kind, offers a robust and fruitful framework for posing and approaching AZD3965 mw scientific and conceptual questions about living systems. It will be shown that functionalism preserves our intuitions about living systems “as-we-know-them”, while providing a strong theoretic framework for encountering and identifying new and novel realizations of living systems in a variety of non-terrestrial physical contexts. Cleland, Carol E., and Christopher F. Chyba. “Defining ‘Life’” Origins of Life and Evolution in the Biosphere 32 (2002): 387–393. Pattee, H. H. “Simulations, Realizations, and Theories of Life.” The Philosophy of Artificial Life. Ed. Margaret A. Boden. New York: Oxford UP, 1996. 379–393. Quine, W.V. O.

The carboxy terminus of CpcA contained a region similar to the ba

The carboxy terminus of CpcA contained a region similar to the basic region of bZIP superfamily of transcription factors with strong sequence similarity to that of the homolog in A. Lonafarnib concentration fumigatus or A. nidulans (Figure 2C). Sapitinib purchase In contrast with the Aspergillus homologs, the leucine zipper region contained three conserved leucine residues characteristic of a leucine zipper L-x(6)-L-x(6)-L-x(6)-L (Figure 2C). As expected for a protein with a transcription factor domain, CpcA was predicted by PSORT II to be localised in the nucleus (69.6% probability) and SignalP did not predict the presence of an N-terminal signal peptide (98.7% probability). In A. nidulans, cpcA transcription is autoregulated via cross pathway

regulatory elements (CPRE) 5′ TGA-(C/G)-TCA-3′ in the cpcA promoter [13]. Point mutations in CPRE lead to low levels of cpcA transcripts and CpcA protein, when amino acids are limited. Such an element matching the consensus was present on the minus strand in the promoter region of L. maculans cpcA (-698 to -703). Figure 2 A) The cpcA locus of Leptosphaeria maculans. The conserved leucine zipper region at the C-terminus of cpcA is dark grey. The open boxes indicate the upstream Open Reading Frames uORF1 and uORF2 in the 5′ leader region. The black dot preceding them represents the putative cross-pathway control element (CPRE) whose sequence is 5′TGACTCA3′. B) Alignment

of the deduced amino acid sequence of uORF2 with counterparts in the leader sequences of Aspergillus Selleck FHPI fumigatus (Af) cpcA (GenBank XP_751584.1) and A. nidulans (An) cpcA (GenBank

AF302935). Black boxes with white text denote amino acids identical in two of the three fungal species. Grey boxes with white check text mark conserved changes. Gaps are introduced to optimize alignment. C) Alignment of the deduced amino acid sequence of the C-terminus conserved leucine zipper region with that of A. fumigatus CpcA and A. nidulans CpcA. The thick black line denotes the bZIP transcription factors basic domain signature (PS00036). Asterisks mark positions where conserved leucine residues characteristic of a leucine zipper (L-x(6)-L-x(6)-L-x(6)-L) should be found. Role of CpcA in sirodesmin PL production in L. maculans Although insertion of the T-DNA downstream of cpcA in mutant GTA7 reduced the transcript size by 127 bp, it did not reduce transcript levels of cpcA compared to those of the wild type (data not shown). Since the efficiency of gene disruption in L. maculans is very low, RNA mediated silencing was exploited to develop an isolate with extremely low levels of cpcA transcripts in order to study the effect of cpcA on sirodesmin PL production. Several putatively-silenced transformants were analysed and one, cpcA-sil, with 10% transcript level of that in wild type, as seen by q RT-PCR analysis, was chosen for further analysis (data not shown).

However, our experimental results contradict the anticipation Th

However, our experimental results contradict the anticipation. The phenomenon can be ascribed to the compensation by the increase of their diameter. Based on our experimental results, the growth time plays an important role in density and selleckchem morphology control of ZnO NWs and thus modifies the optoelectronic

properties for versatile devices. Conclusions In summary, the vertical arrays of well-aligned c-axis orientation ZnO NWs have been synthesized on silicon substrate by VS growth mechanism at a relatively low growth temperature. By varying the growth time, we can adjust the areal density, length, and diameter of ZnO NWs and modify the structural and optoelectronic properties accordingly. PL spectra measured at room temperature exhibit a sharp UV peak and broad green Osimertinib band, click here corresponding to the NBE and defect-related emissions, respectively. When the growth time increased, the average diameter of NWs became larger and thus the surface-to-volume ratio became lower. Therefore, higher surface states of ZnO NWs with smaller diameters can be

responsible for the origin of enhanced green emission. ZnO NWs with strong alignment and uniform distribution can also minimize the reflectance to 5.7% in the visible region. In addition, field emission features revealed that the growth time plays an important role in density- and morphology-controlled ZnO NWs. It is reasonable to expect that the ZnO NWs can be modified to meet the requirements for versatile optoelectronic devices. Acknowledgements This work was supported by the Green Technology Research Center of Chang Gung University and the National Science Council (NSC) of Thalidomide Taiwan under contract numbers NSC100-2815-C-155-013-E, NSC100-2112-M-182-004,

and NSC101-2112-M-182-003-MY3. References 1. Jiang CY, Sun XW, Lo GQ, Kwong DL, Wang JX: Improved dye-sensitized solar cells with a ZnO-nanoflower photoanode. Appl Phys Lett 2007, 90:263501.CrossRef 2. Xu L, Shen H, Li X, Zhu R: Enhanced ultraviolet emission from ZnO thin film covered by TiO2 nanoparticles. Chin Opt Lett 2009, 7:953–955.CrossRef 3. Huang MH, Mao S, Feick H, Yan H, Wu Y, Kind H, Weber E, Russo R, Yang P: Room-temperature ultraviolet nanowire nanolasers. Science 1897, 2001:292. 4. Manoharan MP, Desai AV, Neely G, Haque MA: Synthesis and elastic characterization of zinc oxide nanowires. J Nanomater 2008, 2008:849745.CrossRef 5. Ng HT, Han J, Yamada T, Nguyen P, Chen YP, Meyyappan M: Single crystal nanowire vertical surround-gate field-effect transistor. Nano Lett 2004, 4:1247.CrossRef 6. Heo YW, Tien LC, Kwon Y, Norton DP, Pearton SJ, Kang BS, Ren F: Depletion-mode ZnO nanowire field-effect transistor. Appl Phys Lett 2004, 85:2274.CrossRef 7. Lee CH, Yoo J, Doh YJ, Yi GC: ZnO/Mg 0.2 Zn 0.8 O coaxial nanorod heterostructures for high-performance electronic nanodevice applications. Appl Phys Lett 2009, 94:043504.CrossRef 8. Li QH, Liang YX, Wan Q, Wang TH: Oxygen sensing characteristics of individual ZnO nanowire transistors.

The labeled cRNAs were purified with the RNeasy Mini kit (Qiagen,

The labeled cRNAs were purified with the RNeasy Mini kit (Qiagen, Hilden, Germany) and quantified using NanoDrop ND-1000 UV-VIS spectrophotometer. Aliquots (600 ng) of Cy3-labeled cRNAs were fragmented and hybridized for 17 h at 65°C to each array using the Gene Expression Hybridization see more kit (Agilent Technologies) and according to the manufacturer’s instructions. Microarray imaging and data analysis Slides were washed and processed according to the Agilent 60-mer Oligo Microarray

Processing protocol and scanned on a Agilent microarray scanner G2565BA (Agilent Technologies). Data were extracted from the images with Feature Extraction (FE) software (Agilent Technologies). FE software flags outlier features, and detects and removes spatial gradients and local backgrounds. Data were normalized using a combined rank consistency

filtering with LOWESS intensity normalization. The gene expression values obtained from FE software were imported into GeneSpring 10.0.2 software (Agilent Technologies) for pre-processing and data analysis. For inter-array comparisons, a linear scaling of the data was performed using the 75th percentile signal value of all of non-control probes on the microarray to normalize one-colour signal values. Probe sets with a signal intensity value below the 20th percentile were considered as absent and discarded from subsequent analysis. The expression of each gene was normalized by its median expression across all samples. Genes were included in the final data set if their expression changed by at least twofold between strain H99 FLC-exposed or -not exposed (control sample) in ABT 888 at least two independent experiments, together Phospholipase D1 with a P-value cut-off of < 0.05 (by one-way analysis of variance [ANOVA] corrected). Genes listed in Table 1 were categorized by reported or putative functions by the BROAD Institute database with NCBI blastP http://​www.​ncbi.​nlm.​nih.​gov/​BLAST/​ editing, and also by the Uniprot http://​www.​uniprot.​org/​ and Saccharomyces

genome http://​www.​yeastgenome.​org/​cgi-bin/​blast-sgd.​pl databases. As indicated in Table 1, each S. cerevisiae gene name was assigned by blastP search with the C. neoformans H99 gene sequence (e-value cutoff: e-6) according to Kim et al. [24]. Gene Ontology (GO) term analysis was carried to help categorize a list of genes into functional groups. The whole microarray data have been deposited in National Center for Biotechnology Information’s Gene Expression Omnibus [25] and are accessible through GEO Series accession number GSE24927. Table 1 Changes in the gene expression of C. neoformans H99 cells exposed to FLC BROAD ID (CNAG_*****) C. n. gene name S. c. gene name Description Fold change Ergosterol biosynthesis 04804 SRE1   Sterol regulatory element-binding protein 1 + 4.04 01737   ERG25 C-4 methyl sterol oxidase + 3.95 00854   ERG2 C-8 sterol isomerase + 3.