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The glucose analog 2-deoxyglucose (2DG) inhibits the growth of Saccharomyces cerevisiae and human tumor cells, but its modes of action have not been fully elucidated. Yeast cells lacking Snf1 (AMP-activated protein kinase) are hypersensitive to 2DG. Overexpression of either of two low-affinity, high-capacity glucose transporters, Hxt1 and Hxt3, suppresses the 2DG hypersensitivity of snf1Δ cells. The addition of 2DG or the loss of Snf1 reduces HXT1 and HXT3 expression levels and stimulates transporter endocytosis and degradation in the vacuole. 2DG-stimulated trafficking of Hxt1 and Hxt3 requires Rod1/Art4 and Rog3/Art7, two members of the α-arrestin trafficking adaptor family. Mutations in ROD1 and ROG3 that block binding to the ubiquitin ligase Rsp5 eliminate Rod1- and Rog3-mediated trafficking of Hxt1 and Hxt3. Genetic analysis suggests that Snf1 negatively regulates both Rod1 and Rog3, but via different mechanisms. Snf1 activated by 2DG phosphorylates Rod1 but fails to phosphorylate other known targets, such as the transcriptional repressor Mig1. We propose a novel mechanism for 2DG-induced toxicity whereby 2DG stimulates the modification of α-arrestins, which promote glucose transporter internalization and degradation, causing glucose starvation even when cells are in a glucose-rich environment. [ABSTRACT FROM AUTHOR]

Mip1 is the Saccharomyces cerevisiae DNA polymerase γ (Pol γ), which is responsible for the replication of mitochondrial DNA (mtDNA). It belongs to the family A of the DNA polymerases and it is orthologs to human POLGA. In humans, mutations in POLG(1) cause many mitochondrial pathologies, such as progressive external ophthalmoplegia (PEO), Alpers' syndrome, and ataxia-neuropathy syndrome, all of which present instability of mtDNA, which results in impaired mitochondrial function in several tissues with variable degrees of severity. In this review, we summarize the genetic and biochemical knowledge published on yeast mitochondrial DNA polymerase from 1989, when the MIP1 gene was first cloned, up until now. The role of yeast is particularly emphasized in (i) validating the pathological mutations found in human POLG and modeled in MIP1, (ii) determining the molecular defects caused by these mutations and (iii) finding the correlation between mutations/polymorphisms in POLGA and mtDNA toxicity induced by specific drugs. We also describe recent findings regarding the discovery of molecules able to rescue the phenotypic defects caused by pathological mutations in Mip1, and the construction of a model system in which the human Pol γ holoenzyme is expressed in yeast and complements the loss of Mip1. [ABSTRACT FROM AUTHOR]

Copyright of Turkish Bulletin of Hygiene & Experimental Biology / Türk Hijyen ve Deneysel Biyoloji is the property of Refik Saydam National Public Health Agency and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In eukaryotic cells, local chromatin structure and chromatin organization in the nucleus both influence transcriptional regulation. At the local level, the Fun30 chromatin remodeler Fft3 is essential for maintaining proper chromatin structure at centromeres and subtelomeres in fission yeast. Using genome-wide mapping and live cell imaging, we show that this role is linked to controlling nuclear organization of its targets. In fft3∆ cells, subtelomeres lose their association with the LEM domain protein Man1 at the nuclear periphery and move to the interior of the nucleus. Furthermore, genes in these domains are upregulated and active chromatin marks increase. Fft3 is also enriched at retrotransposon-derived long terminal repeat (LTR) elements and at tRNA genes. In cells lacking Fft3, these sites lose their peripheral positioning and show reduced nucleosome occupancy. We propose that Fft3 has a global role in mediating association between specific chromatin domains and the nuclear envelope. [ABSTRACT FROM AUTHOR]

Although numerous regulatory connections between pre-mRNA splicing and chromatin have been demonstrated, the precise mechanisms by which chromatin factors influence spliceosome assembly and/or catalysis remain unclear. To probe the genetic network of pre-mRNA splicing in the fission yeast Schizosaccharomyces pombe, we constructed an epistatic mini-array profile (E-MAP) and discovered many new connections between chromatin and splicing. Notably, the nucleosome remodeler SWI/SNF had strong genetic interactions with components of the U2 snRNP SF3 complex. Overexpression of SF3 components in ΔSWI/SNF cells led to inefficient splicing of many fission yeast introns, predominantly those with non-consensus splice sites. Deletion of SWI/SNF decreased recruitment of the splicing ATPase Prp2, suggesting that SWI/SNF promotes co-transcriptional spliceosome assembly prior to first step catalysis. Importantly, defects in SWI/SNF as well as SF3 overexpression each altered nucleosome occupancy along intron-containing genes, illustrating that the chromatin landscape both affects—and is affected by—co-transcriptional splicing. [ABSTRACT FROM AUTHOR]

Separase/Esp1 is a protease required at the onset of anaphase to cleave cohesin and thereby enable sister chromatid separation. Esp1 also promotes release of the Cdc14 phosphatase from the nucleolus to enable mitotic exit. To uncover other potential roles for separase, we performed two complementary genome-wide genetic interaction screens with a strain carrying the budding yeast esp1-1 separase mutation. We identified 161 genes that when mutated aggravate esp1-1 growth and 44 genes that upon increased dosage are detrimental to esp1-1 viability. In addition to the expected cell cycle and sister chromatid segregation genes that were identified, 24% of the genes identified in the esp1-1 genetic screens have a role in Ty1 element retrotransposition. Retrotransposons, like retroviruses, replicate through reverse transcription of an mRNA intermediate and the resultant cDNA product is integrated into the genome by a conserved transposon or retrovirus encoded integrase protein. We purified Esp1 from yeast and identified an interaction between Esp1 and Ty1 integrase using mass spectrometry that was subsequently confirmed by co-immunoprecipitation analysis. Ty1 transposon mobility and insertion upstream of the SUF16 tRNA gene are both reduced in an esp1-1 strain but increased in cohesin mutant strains. Securin/Pds1, which is required for efficient localization of Esp1 to the nucleus, is also required for efficient Ty1 transposition. We propose that Esp1 serves two roles to mediate Ty1 transposition – one to remove cohesin and the second to target Ty1-IN to chromatin. [ABSTRACT FROM AUTHOR]

Patulin contamination of fruit- and vegetable-based products had become a major challenge for the food industry. Biological methods of patulin control can play an important role due to their safety and high efficiency. In this study, a strain of marine yeast with high patulin degradation ability was screened. The yeast was identified asKodameae ohmeriby the BioLog identification system and partial 26S rRNA gene sequencing. The degradation products of patulin were identified as (E)- and (Z)-ascladiol through HPLC and LC-TOF/MS. High patulin tolerance at 100 μg ml–1and a high degradation rate at 35°C at a pH between 3 and 6 indicates the potential application ofK. ohmerifor patulin detoxification of apple-derived products. [ABSTRACT FROM PUBLISHER]

Complex sphingolipids play important roles in many physiologically important events in yeast S accharomyces cerevisiae. In this study, we screened yeast mutant strains showing a synthetic lethal interaction with loss of mannosylinositol phosphorylceramide ( MIPC) synthesis and found that a specific group of glycosyltransferases involved in the synthesis of mannan-type N-glycans is essential for the growth of cells lacking MIPC synthases ( Sur1 and Csh1). The genetic interaction was also confirmed by repression of MNN2, which encodes alpha-1,2-mannosyltransferase that synthesizes mannan-type N-glycans, by a tetracycline-regulatable system. MNN2-repressed sur1Δ csh1Δ cells exhibited high sensitivity to zymolyase treatment, and caffeine and sodium dodecyl sulfate ( SDS) strongly inhibited the growth of sur1Δ csh1Δ cells, suggesting impairment of cell integrity due to the loss of MIPC synthesis. The phosphorylated form of Slt2, a mitogen-activated protein ( MAP) kinase activated by impaired cell integrity, increased in sur1Δ csh1Δ cells, and this increase was dramatically enhanced by the repression of Mnn2. Moreover, the growth defect of MNN2-repressed sur1Δ csh1Δ cells was enhanced by the deletion of SLT2 or RLM1 encoding a downstream target of Slt2. These results indicated that loss of MIPC synthesis causes impairment of cell integrity, and this effect is enhanced by impaired synthesis of mannan-type N-glycans. [ABSTRACT FROM AUTHOR]

The year-round dynamics of the number and taxonomic composition of yeast communities in the soddy-podzolic soils under invasive thickets of Heracleum sosnowskyi were investigated. The yeast groups that are formed in the soil under the continuous Sosnowsky's hogweed thickets significantly differ from the indigenous yeast communities under the adjacent meadows. In the soils of both biotopes, typical eurybiotic yeast species predominate. In the soil under Heracleum sosnowskyi, the share of the ascomycetes Candida vartiovaarae and Wickerhamomyces anomalus is much lower, and the portion of yeast-like fungi with high hydrolytic activity such as Trichosporon moniliforme and Trichosporon porosum is greater. A possible explanation for this phenomenon is that Sosnowsky's hogweed, unlike most aboriginal meadow grasses, does not hibernate with green leaves that do not gradually die out with the formation of semidecomposed plant residues-the main source of nutrients for the soil-litter microbial complex. In addition, grasses of the lower layer do not develop under Sosnowsky's hogweed due to the strong shading and allelopathic impact preventing the development of typical epiphytic copiotrophic species of yeasts. [ABSTRACT FROM AUTHOR]

The article presents a study examining how the fungus Neurospora crassa breaks down hemicellulose and suggesting that giving years the ability to break down hemicellulose may enhance the efficiency of biofuel production. It describes the protein identified in the study that can transport molecules of xylodextrin into the cells of the fungus, as well as the two enzymes that break down the xylodextrin to make simple sugars.

The process of cocoa fermentation is a very important step for the generation or aromatic compounds, which are attributable to the metabolism of the microorganisms involved. There are some reports about this process and the identification of microorganisms; however, there are no reports identifying the yeasts involved in a Mexican cocoa fermentation process using molecular biology techniques, including restricted fragment length polymorphism (RFLP) and denaturing gradient gel electrophoresis (DGGE). The aim of this study was to identify the main yeast species associated with Mexican cocoa fermentations employing culture-dependent and -independent techniques achieving two samplings with a 1 year time difference at the same site. Isolation of the microorganisms was performed in situ. Molecular identification of yeast isolates was achieved by RFLP analysis and rDNA sequencing. Total DNA from the microorganisms on the cocoa beans was utilized for the DGGE analysis. Bands from the DGGE gels were excised and sequenced. Nineteen isolated yeasts were identified (al specie level), three of which had never before been associated with cocoa fermentations worldwide. The detected predominant yeast varied from one technique to another. Hanseniaspora sp. resulted dominant in DGGE however Saccharomyces cerevisiae was the principal isolated species. In conclusion, the culture-dependent and -independent techniques complement each other showing differences in the main yeasts involved in spontaneous cocoa fermentation, probably due to the physiological states of the viable but non culturable yeasts. Furthermore important differences between the species detected in the two samplings were detected. [ABSTRACT FROM AUTHOR]

Background: The histone variant H2A.Z, which has been reported to have both activating and repressive effects on gene expression, is known to occupy nucleosomes at the 5' ends of protein-coding genes. Results: We now find that H2A.Z is also significantly enriched in gene coding regions and at the 3' ends of genes in budding yeast, where it co-localises with histone marks associated with active promoters. By comparing H2A.Z binding to global gene expression in budding yeast strains engineered so that normally unstable transcripts are abundant, we show that H2A.Z is required for normal levels of antisense transcripts as well as sense ones. High levels of H2A.Z at antisense promoters are associated with decreased antisense transcript levels when H2A.Z is deleted, indicating that H2A.Z has an activating effect on antisense transcripts. Decreases in antisense transcripts affected by H2A.Z are accompanied by increased levels of paired sense transcripts. Conclusions: The effect of H2A.Z on protein coding gene expression is a reflection of its importance for normal levels of both sense and antisense transcripts. [ABSTRACT FROM AUTHOR]

Yeast cells are protected by a cell wall that plays an important role in the exchange of substances with the environment. The cell wall structure is dynamic and can adapt to different physiological states or environmental conditions. For the investigation of morphological changes, selective staining with fluorescent dyes is a valuable tool. Furthermore, cell wall staining is used to facilitate sub-cellular localization experiments with fluorescently-labeled proteins and the detection of yeast cells in non-fungal host tissues. Here, we report staining of Saccharomyces cerevisiae cell wall with Trypan Blue, which emits strong red fluorescence upon binding to chitin and yeast glucan; thereby, it facilitates cell wall analysis by confocal and super-resolution microscopy. The staining pattern of Trypan Blue was similar to that of the widely used UV-excitable, blue fluorescent cell wall stain Calcofluor White. Trypan Blue staining facilitated quantification of cell size and cell wall volume when utilizing the optical sectioning capacity of a confocal microscope. This enabled the quantification of morphological changes during growth under anaerobic conditions and in the presence of chemicals, demonstrating the potential of this approach for morphological investigations or screening assays. [ABSTRACT FROM AUTHOR]

Traditionally, high-value metabolites have been produced through direct extraction from natural biological sources which are inefficient, given the low abundance of these compounds. On the other hand, these high-value metabolites are usually difficult to be synthesized chemically, due to their complex structures. In the last few years, the discovery of genes involved in the synthetic pathways of these metabolites, combined with advances in synthetic biology tools, has allowed the construction of increasing numbers of yeast cell factories for production of these metabolites from renewable biomass. This review summarizes recent advances in synthetic biology in terms of the use of yeasts as microbial hosts for the identification of the pathways involved in the synthesis, as well as for the production of high-value metabolites. [ABSTRACT FROM AUTHOR]

The top-view, two-dimensional spatial patterning of non-uniform growth in a Saccharomyces cerevisiae yeast colony is considered. Experimental images are processed to obtain data sets that provide spatial information on the cell-area that is occupied by the colony. A method is developed that allows for the analysis of the spatial distribution with three metrics. The growth of the colony is quantified in both the radial direction from the centre of the colony and in the angular direction in a prescribed outer region of the colony. It is shown that during the period of 100–200 hours from the start of the growth of the colony there is an increasing amount of non-uniform growth. The statistical framework outlined in this work provides a platform for comparative quantitative assays of strain-specific mechanisms, with potential implementation in inferencing algorithms used for parameter-rate estimation. [ABSTRACT FROM AUTHOR]

The present investigation was aimed at understanding the molecular mechanism of gene amplification. Interplay of fragile sites in promoting gene amplification was also elucidated. The amplification promoting sequences were chosen from the Saccharomyces cerevisiae ARS, 5S rRNA regions of Plantago ovata and P. lagopus, proposed sites of replication pausing at Ste20 gene locus of S. cerevisiae, and the bend DNA sequences within fragile site FRA11A in humans. The gene amplification assays showed that plasmid bearing APS from yeast and human beings led to enhanced protein concentration as compared to the wild type. Both the in silico and in vitro analyses were pointed out at the strong bending potential of these APS. In addition, high mitotic stability and presence of TTTT repeats and SAR amongst these sequences encourage gene amplification. Phylogenetic analysis of S. cerevisiae ARS was also conducted. The combinatorial power of different aspects of APS analyzed in the present investigation was harnessed to reach a consensus about the factors which stimulate gene expression, in presence of these sequences. It was concluded that the mechanism of gene amplification was that AT rich tracts present in fragile sites of yeast serve as binding sites for MAR/SAR and DNA unwinding elements. The DNA protein interactions necessary for ORC activation are facilitated by DNA bending. These specific bindings at ORC promote repeated rounds of DNA replication leading to gene amplification. [ABSTRACT FROM AUTHOR]

Yeasts, which have been a component of the human diet for at least 7,000 years, possess an elaborate cell wall α-mannan. The influence of yeast mannan on the ecology of the human microbiota is unknown. Here we show that yeast α-mannan is a viable food source for the Gram-negative bacterium Bacteroides thetaiotaomicron, a dominant member of the microbiota. Detailed biochemical analysis and targeted gene disruption studies support a model whereby limited cleavage of α-mannan on the surface generates large oligosaccharides that are subsequently depolymerized to mannose by the action of periplasmic enzymes. Co-culturing studies showed that metabolism of yeast mannan by B. thetaiotaomicron presents a 'selfish' model for the catabolism of this difficult to breakdown polysaccharide. Genomic comparison with B. thetaiotaomicron in conjunction with cell culture studies show that a cohort of highly successful members of the microbiota has evolved to consume sterically-restricted yeast glycans, an adaptation that may reflect the incorporation of eukaryotic microorganisms into the human diet. [ABSTRACT FROM AUTHOR]

Calorie-restriction extends lifespan in many multicellular organisms; here substances secreted by calorie-restricted yeast are found to induce longer life in other yeast cells, suggesting that cellular communication is a component of this phenomenon even in a single-celled organism. [ABSTRACT FROM AUTHOR]

Manganese (Mn) toxicity is a major constraint limiting plant growth on acidic soils. Superior Mn tolerance in Stylosanthes spp. has been well documented, but its molecular mechanisms remain largely unknown. In this study, superior Mn tolerance in Stylosanthes guianensis was confirmed, as reflected by a high Mn toxicity threshold. Furthermore, genetic variation of Mn tolerance was evaluated using two S. guianensis genotypes, which revealed that the Fine-stem genotype had higher Mn tolerance than the TPRC2001-1 genotype, as exhibited through less reduction in dry weight under excess Mn, and accompanied by lower internal Mn concentrations. Interestingly, Mn-stimulated increases in malate concentrations and exudation rates were observed only in the Fine-stem genotype. Proteomic analysis of Fine-stem roots revealed that S. guianensis Malate Dehydrogenase1 (SgMDH1) accumulated in response to Mn toxicity. Western-blot and quantitative PCR analyses showed that Mn toxicity resulted in increased SgMDH1 accumulation only in Fine-stem roots, but not in TPRC2001-1. The function of SgMDH1-mediated malate synthesis was verified through in vitro biochemical analysis of SgMDH1 activities against oxaloacetate, as well as in vivo increased malate concentrations in yeast (Saccharomyces cerevisiae), soybean (Glycine max) hairy roots, and Arabidopsis (Arabidopsis thaliana) with SgMDH1 overexpression. Furthermore, SgMDH1 overexpression conferred Mn tolerance in Arabidopsis, which was accompanied by increased malate exudation and reduced plant Mn concentrations, suggesting that secreted malate could alleviate Mn toxicity in plants. Taken together, we conclude that the superior Mn tolerance of S. guianensis is achieved by coordination of internal and external Mn detoxification through malate synthesis and exudation, which is regulated by SgMDH1 at both transcription and protein levels. [ABSTRACT FROM AUTHOR]

Toxic effects of Bonny light crude oil on the growth of three aquatic yeasts namely Yarrowia lipolytica, Candida tropicalis and Debryomyces hansenii were studied based on their biomass and protein production. The species showed different responses to the toxic influences of various crude oil concentrations. The growth response was measured spectrophotometrically using optical density (OD) at 600nm. Yarrowia lipolytica responded positively to different crude oil levels. A general assessment indicated that 2% (v/v) crude oil concentration stimulated maximum growth and protein production of this organism. Lower yields were observed at reduced crude oil levels. Growth decreased gradually among Candida tropicalis and Debryomyces hansenii cultures in comparison to the control. Biomass of Candida tropicalis increased from 0.1 (OD600nm) at 0 h to 0.49 after 20 h at 0.5% crude oil concentration. This level gradually declined to 0.04 after 20 h cultivation at 1.5% crude oil concentration. Maximum decline in optical density of this organism was observed at crude oil concentration of 2.0%. Protein levels for Candida tropicalis decreased from 0.13 mg/mL after 20 h at crude oil concentration of 0.5% to 0.04 mg/mL after 20 h at maximum crude oil concentration of 2%. The biomass of Debryomyces hansenii increased slightly from 0.1(OD600nm) at 0 h to 0.44 after 20 h at 0.5% crude oil level. Further decreases in OD values of this organism occurred progressively as the crude oil concentration was increased. Lowest protein yield was observed at a crude oil concentration of 2% at which the least protein production of 0.05 mg/mL was produced after 20 h. [ABSTRACT FROM AUTHOR]

A tripodal Schiff base tris[2-(5-bromosalicylideneamino)ethyl]amine (HL) was prepared by the reaction of 5-bromosalicylaldehyde with tris(2-aminoethyl)amine. Reaction with the Schiff base with manganese perchlorate in methanol resulted a mononuclear manganese(III) complex ( I). The crystal structures of the Schiff base and the complex have been determined by single crystal X-ray diffraction (CIF files CCDC nos. 1007902 (HL); 1007983 ( I)). The Schiff base coordinates to the Mn atom through all the phenolate O and imino N atoms. The Mn atom of the complex is in octahedral coordination. The antibacterial properties have been tested on some bacteria and yeast. [ABSTRACT FROM AUTHOR]

The mechanisms by which RNA-binding proteins control the translation of subsets of mRNAs are not yet clear. Slf1p and Sro9p are atypical-La motif containing proteins which are members of a superfamily of RNA-binding proteins conserved in eukaryotes. RIP-Seq analysis of these two yeast proteins identified overlapping and distinct sets of mRNA targets, including highly translated mRNAs such as those encoding ribosomal proteins. In paralell, transcriptome analysis of slf1Δ and sro9Δ mutant strains indicated altered gene expression in similar functional classes of mRNAs following loss of each factor. The loss of SLF1 had a greater impact on the transcriptome, and in particular, revealed changes in genes involved in the oxidative stress response. slf1Δ cells are more sensitive to oxidants and RIP-Seq analysis of oxidatively stressed cells enriched Slf1p targets encoding antioxidants and other proteins required for oxidant tolerance. To quantify these effects at the protein level, we used label-free mass spectrometry to compare the proteomes of wild-type and slf1Δ strains following oxidative stress. This analysis identified several proteins which are normally induced in response to hydrogen peroxide, but where this increase is attenuated in the slf1Δ mutant. Importantly, a significant number of the mRNAs encoding these targets were also identified as Slf1p-mRNA targets. We show that Slf1p remains associated with the few translating ribosomes following hydrogen peroxide stress and that Slf1p co-immunoprecipitates ribosomes and members of the eIF4E/eIF4G/Pab1p ‘closed loop’ complex suggesting that Slf1p interacts with actively translated mRNAs following stress. Finally, mutational analysis of SLF1 revealed a novel ribosome interacting domain in Slf1p, independent of its RNA binding La-motif. Together, our results indicate that Slf1p mediates a translational response to oxidative stress via mRNA-specific translational control. [ABSTRACT FROM AUTHOR]

During exocytosis, the evolutionarily conserved exocyst complex tethers Golgi-derived vesicles to the target plasma membrane, a critical function for secretory pathways. Here we show that exo70B1 loss-of-function mutants express activated defense responses upon infection and express enhanced resistance to fungal, oomycete and bacterial pathogens. In a screen for mutants that suppress exo70B1 resistance, we identified nine alleles of TIR-NBS2 (TN2), suggesting that loss-of-function of EXO70B1 leads to activation of this nucleotide binding domain and leucine-rich repeat-containing (NLR)-like disease resistance protein. This NLR-like protein is atypical because it lacks the LRR domain common in typical NLR receptors. In addition, we show that TN2 interacts with EXO70B1 in yeast and in planta. Our study thus provides a link between the exocyst complex and the function of a ‘TIR-NBS only’ immune receptor like protein. Our data are consistent with a speculative model wherein pathogen effectors could evolve to target EXO70B1 to manipulate plant secretion machinery. TN2 could monitor EXO70B1 integrity as part of an immune receptor complex. [ABSTRACT FROM AUTHOR]

Abscission is the final step of cytokinesis that involves the cleavage of the intercellular bridge connecting the two daughter cells. Recent studies have given novel insight into the spatiotemporal regulation and molecular mechanisms controlling abscission in cultured yeast and human cells. The mechanisms of abscission in living metazoan tissues are however not well understood. Here we show that ALIX and the ESCRT-III component Shrub are required for completion of abscission during Drosophila female germline stem cell (fGSC) division. Loss of ALIX or Shrub function in fGSCs leads to delayed abscission and the consequent formation of stem cysts in which chains of daughter cells remain interconnected to the fGSC via midbody rings and fusome. We demonstrate that ALIX and Shrub interact and that they co-localize at midbody rings and midbodies during cytokinetic abscission in fGSCs. Mechanistically, we show that the direct interaction between ALIX and Shrub is required to ensure cytokinesis completion with normal kinetics in fGSCs. We conclude that ALIX and ESCRT-III coordinately control abscission in Drosophila fGSCs and that their complex formation is required for accurate abscission timing in GSCs in vivo. [ABSTRACT FROM AUTHOR]

The monitoring and control of bioprocesses is a challenging task. This applies particularly if the actions to the process have to be carried out in real-time. This work presents a system for on-line monitoring and control of batch yeast propagation under limiting conditions based on a virtual plant operator, which uses the concept of intelligent control algorithms by means of fuzzy logic theory. Process information is provided on-line using a sensor array comprising the measurement of OD, operating temperature, pressure, density, dissolved oxygen, and pH value. In this context practical problems arising through on-line sensing and signal processing are addressed. The preprocessed sensor data are fed to a neural network for on-line biomass estimation. The root mean squared error of prediction is 4 × 106 cells/mL. The proposed system then triggers temperature and aeration by usage of a temperature dependent metabolic growth model and sensor data. The deviation of the predicted biomass from that of the reference trajectory as modeled by the metabolic growth model and its temporal derivative are used as inputs for the fuzzy temperature controller. The inputs used by the fuzzy aeration controller are the deviation of measured extract from that of the reference trajectory, the predicted cell count, and the dissolved oxygen concentration. The fuzzy-based expert system allows to provide the desired yeast cell concentration of 100-120 × 106 cells/mL at a minimum residual extract limit of 6.0 g/100 g at the required point of time. Thus, a dynamic adjustment of the propagation process to the overall production schedule is possible in order to produce the required amount of biomass at the right time. [ABSTRACT FROM AUTHOR]

Background Infancy is a developmental stage with heightened susceptibility to environmental influences on the risk of chronic childhood disease. Few birth cohort studies have detailed measures of fungal diversity data in infants' bedrooms, limiting the potential to measure long-term associations of these complex exposures with development of asthma or allergy. Objective We evaluated the relation of home fungal levels in infancy to repeated measures of wheeze and development of asthma and rhinitis by age 13, and sensitization by age 12 years. Methods In the Epidemiology of Home Allergens and Asthma prospective birth cohort study, we recruited 408 children with family history of allergic disease or asthma. When children were aged 2-3 months, we measured culturable fungi in bedroom air and dust, and in outdoor air. Main outcomes included ascertainment of symptoms/disease onset by questionnaire from birth through age 13. We estimated hazard ratios and, for wheeze and sensitization, odds ratios for an interquartile increase in log-transformed fungal concentrations, adjusting for other outcome predictors and potential confounders. Results Elevated levels of yeasts in bedroom floor dust were associated with reduced: i) wheeze at any age; ii) fungal sensitization; and iii) asthma development by age 13 (hazard ratio ( HR) = 0.86; 95% confidence interval ( CI), [0.75 to 0.98]). Outdoor airborne Cladosporium and dustborne Aspergillus predicted increased rhinitis. Risk of fungal sensitization by age 12, in response to environmental Alternaria and Aspergillus, was elevated in children with a maternal history of fungal sensitization. Conclusions and Clinical Relevance Despite the irritant and allergenic properties of fungi, early-life elevated dust yeast exposures or their components may be protective against allergy and asthma in children at risk for these outcomes. Ascertainment of fungal components associated with immunoprotective effects may have therapeutic relevance for asthma. [ABSTRACT FROM AUTHOR]

Phosphatidate phosphatase-1 (PAP1) enzymes (yeast Pah1p/Smp2p, mammalian lipin1-3) have a key role in lipid homeostasis by controlling the relative proportions of its substrate phosphatidate (PA) and its product diacylglycerol (DAG). Recent investigation shows that mammalian lipin-1 complements phenotypes exhibited by yeast pah1Δ mutant cells, which indicates the functions of PAP1 enzymes are evolutionarily conserved. The observation was confirmed after transformation of human LPIN1 into PAH1-defective yeast, which resulted in human LPIN1-induced accumulation of triacylglycerol (TAG )and lipid droplet formation. In double mutants lacking Tgl3p and Tgl4p, overexpression of PAH1 or LPIN1 induced TAG accumulation and excessive obesity. Furthermore, the obese yeast was used as a model to study the anti-obesity effects of PAP1 activity inhibitors, including propranolol and clenbuterol. The data showed that the inhibitors significantly suppressed TAG accumulation and lipid droplets formation. These findings demonstrate that LPIN1 plays a functional role in lipid synthesis and storage, a role which is highly conserved from human to yeast. Inhibition of TAG synthesis will become an efficacious treatment strategy for obesity and our excessive obesity model will provide a very useful tool for discovery of new anti-obesity drugs in the future. [ABSTRACT FROM AUTHOR]

Copyright of Revista Colombiana de Biotecnología is the property of Universidad Nacional de Colombia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Proteasomes are conserved protease complexes enriched in the nuclei of dividing yeast cells, a major site for protein degradation. If yeast cells do not proliferate and transit to quiescence, metabolic changes result in the dissociation of proteasomes into proteolytic core and regulatory complexes and their sequestration into motile cytosolic proteasome storage granuli. These granuli rapidly clear with the resumption of growth, releasing the stored proteasomes, which relocalize back to the nucleus to promote cell cycle progression. Here, I report on three models of how proteasomes are transported from the cytoplasm into the nucleus of yeast cells. The first model applies for dividing yeast and is based on the canonical pathway using classical nuclear localization sequences of proteasomal subcomplexes and the classical import receptor importin/karyopherin αβ. The second model applies for quiescent yeast cells, which resume growth and use Blm10, a HEAT-like repeat protein structurally related to karyopherin β, for nuclear import of proteasome core particles. In the third model, the fully-assembled proteasome is imported into the nucleus. Our still marginal knowledge about proteasome dynamics will inspire the discussion on how protein degradation by proteasomes may be regulated in different cellular compartments of dividing and quiescent eukaryotic cells. [ABSTRACT FROM AUTHOR]

We evaluated the effects of yeast polysaccharide (YSP) on the immune function of Apostichopus japonicus exposed to a decrease in salinity (3, 5, or 9 ppt/day). Juvenile sea cucumbers were fed artificial feed containing 0.5 % YSP. We measured phagocytic activity and lysozyme, acid phosphatase, and alkaline phosphatase activity 24 h after each decrease in salinity. When salinity was decreased by 3 ppt/day, after 24 h, the immune index were significantly enhanced by supplementation of the diet with YAP ( P < 0.05). There was no obvious change in the phagocytic index of animals fed a YSP supplemented diet and exposed to a decrease of 5 ppt/day, though the other immune indices were significantly elevated relative to the starting control (30 ppt). The immune index was significantly higher in animals fed a YSP supplemented diet and exposed to a decrease of 9 ppt/day than in animals held at 30 ppt. It is concluded that the addition of YSP to the basal diet improved the immunocompetence of A. japonicas during exposure to the salinity stressors. [ABSTRACT FROM AUTHOR]

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In Greece standard tests are performed in the watering and cooling systems of hotels' units either as part of the surveillance scheme or following human infection. The purpose of this study was to establish the minimum inhibitory concentration (MIC) distributions of environmental Legionella isolates for six antimicrobials commonly used for the treatment of Legionella infections, by MIC-test methodology. Water samples were collected from 2004 to 2011 from 124 hotels from the four prefectures of Crete (Greece). Sixty-eight (68) Legionella isolates, comprising L pneumophila serogroups 1, 2, 3, 5, 6, 8,12,13,15, L anisa, L rubrilucens, L maceachernii, L quinlivanii, L. oakridgensis, and L taurinensis, were included in the study. MIC-tests were performed on buffered charcoal yeast extract with α-ketoglutarate, L-cysteine, and ferric pyrophosphate. The MICs were read after 2 days of incubation at 36 ± 1 WC at 2.5% CO2. A large distribution in MICs was recorded for each species and each antibiotic tested. Rifampicin proved to be the most potent antibiotic regardless of the Legionella spp.; tetracycline appeared to have the least activity on our environmental isolates. The MIC-test approach is an easy, although not so cost-effective, way to determine MICs in Legionella spp. These data should be kept in mind especially since these Legionella species may cause human disease. [ABSTRACT FROM AUTHOR]

Biodiversity of yeast in natural fermented dairy and traditional kefir grain from Inner Mongolia was investigated. The purpose was to explore the main microflora and yeast species, laying the foundation for selecting yeast which can ferment low alcohol degree dairy. Yeast strains were isolated by gradient diluting and streaking plates, and then classified preliminarily according to colony morphology and cell morphology, followed by 5.8S rDNA-ITS restriction fragment length polymorphism (RFLP) analysis, 130 selected yeast strains were divided into 10 phenotypes and 6 molecular types. The species identified by 5.8S ITS-RFLP corresponded to Kluyveromyces marxianus, Cryptococcus sp., Saccharomyces cerevisiae, Candida tropicalis, Issatchenkia terricola and Pichia guilliermondii. Among them, K. marxianus, S. cerevisiae and P. guiltiermondii were the dominant species, which had the potential for fermenting low alcohol degree dairy. [ABSTRACT FROM AUTHOR]

The adjustment of feeding behavior in response to hunger and satiety contributes to homeostatic regulation in animals. The fruit fly Drosophila melanogaster feeds on yeasts growing on overripe fruit, providing nutrients required for adult survival, reproduction and larval growth. Here, we present data on how the nutritional value of food affects subsequent yeast consumption in Drosophila adult males. After a period of starvation, flies showed intensive yeast consumption. In comparison, flies stopped feeding after having access to a nutritive cornmeal diet. Interestingly, dietary glucose was equally efficient as the complex cornmeal diet. In contrast, flies fed with sucralose, a non-metabolizable sweetener, behaved as if they were starved. The adipokinetic hormone and insulin-like peptides regulate metabolic processes in insects. We did not find any effect of the adipokinetic hormone pathway on this modulation. Instead, the insulin pathway was involved in these changes. Flies lacking the insulin receptor (InR) did not respond to nutrient deprivation by increasing yeast consumption. Together these results show the importance of insulin in the regulation of yeast consumption in response to starvation in adult D. melanogaster males. [ABSTRACT FROM AUTHOR]

Chemical sanitizers are usually added to dump tank water to minimize cross-contamination during tomato packing. However, the efficacy of sanitizers continues to be questioned. This study assessed the ability of six commonly used sanitizers (40 ppm of peroxyacetic acid, 40 ppm of mixed peracid, 40 ppm of available chlorine alone or acidified to pH 6.0 with citric acid or T-128, and electrolyzed water containing 40 ppm of available chlorine at pH 6.7) to reduce Salmonella on tomatoes, in wash water, and on equipment surfaces using a pilot-scale processing line. Red round tomatoes (11.3 kg) were dip inoculated to contain Salmonella at ~ 6 log CFU/g, air dried for 2 h, treated for 2 min in a 3.3-m-long dump tank and then dried on a roller conveyor, with sanitizer-free water serving as the control. Tomato and water samples were collected at 15-s intervals during washing with additional dump tank, water tank, and roller conveyor surface samples collected after washing. All samples were appropriately neutralized, diluted, and surface plated on Trypticase soy agar containing 0.6% yeast extract, 0.05% ferric ammonium citrate, and 0.03% sodium thiosulfate with or without membrane filtration to enumerate Salmonella. All six sanitizer treatments were more efficacious than the water control (P ≤ 0.05), with chlorine plus citric acid yielding the greatest Salmonella reduction on tomatoes (3.1 log CFU/g). After processing, all sanitizer wash solutions contained significantly lower (P ≤ 0.05) levels of Salmonella than the water control (3.0 log CFU/ml). The four chlorine-based sanitizer treatments yielded significantly lower Salmonella populations (P ≤ 0.05) in the wash solution compared with peroxyacetic acid and mixed peracid. After processing with sanitizers, Salmonella populations decreased to nondetectable levels (< 0.2 log CFU/100 cm²) on the equipment surfaces. [ABSTRACT FROM AUTHOR]

We present a microfluidic device, which enables single cells to be reliably trapped and cultivated while simultaneously being monitored by means of multifrequency electrical impedance spectroscopy (EIS) in the frequency range of 10 kHz-10 MHz. Polystyrene beads were employed to characterize the EIS performance inside the microfluidic device. The results demonstrate that EIS yields a low coefficient of variation in measuring the diameters of captured beads (~0.13 %). Budding yeast, Saccharomyces cerevisiae, was afterwards used as model organism. Single yeast cells were immobilized and measured by means of EIS. The bud growth was monitored through EIS at a temporal resolution of 1 min. The size increment of the bud, which is difficult to determine optically within a short time period, can be clearly detected through EIS signals. The impedance measurements also reflect the changes in position or motion of single yeast cells in the trap. By analyzing the multifrequency EIS data, cell motion could be qualitatively discerned from bud growth. The results demonstrate that single-cell EIS can be used to monitor cell growth, while also detecting potential cell motion in real-time and label-free approach, and that EIS constitutes a sensitive tool for dynamic single-cell analysis. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

The arrangement of chromatin within interphase nuclei seems to be caused by topological constraints and related to gene expression depending on tissue and developmental stage. In yeast and animals it was found that homologous and heterologous chromatin association are required to realize faithful expression and DNA repair. To test whether such associations are present in plants we analyzed Arabidopsis thaliana interphase nuclei by FISH using probes from different chromosomes. We found that chromatin fiber movement and variable associations, although in general relatively seldom, may occur between euchromatin segments along chromosomes, sometimes even over large distances. The combination of euchromatin segments bearing high or low co-expressing genes did not reveal different association frequencies probably due to adjacent genes of deviating expression patterns. Based on previous data and on FISH analyses presented here, we conclude that the global interphase chromatin organization in A. thaliana is relatively stable, due to the location of its 10 centromeres at the nuclear periphery and of the telomeres mainly at the centrally localized nucleolus. Nevertheless, chromatin movement enables a flexible spatial genome arrangement in plant nuclei. [ABSTRACT FROM AUTHOR]

The environmental isolation of opportunistic pathogenic black yeasts, which are responsible for a wide spectrum of human infections, is essential to understanding the ecology of clinical fungi. Extreme outdoor environments polluted with aromatic hydrocarbons support the growth of black yeasts in unlikely places, such as railway sleepers. However, there are limited data concerning the diversity of these fungi growing on polluted railway sleepers. In this investigation, we examined 845 railway sleeper samples, obtained from 11 Turkish cities representing altitudes from 25 to 1,893 m, and inoculated the samples onto mycological media for the isolation of black yeasts. Ninety-four samples (11.1 %) yielded positive results for black yeast, with creosoted oak sleepers having a significantly higher number of isolates than concrete sleepers ( p < 0.05). Identification based on the ribosomal DNA (rDNA) internal transcribed spacer region revealed the highest prevalence of Exophiala phaeomuriformis, followed by Exophiala dermatitidis, Exophiala heteromorpha, Exophiala xenobiotica, and Exophiala crusticola. This study revealed that railway sleepers harboring black yeasts were predominantly (>75 %) populated with thermophilic species. We observed that altitude might have a significant effect on species diversity. Briefly, E. phaeomuriformis exhibited growth over a wide altitude range, from 30 to 1,893 m. In contrast, E. dermatitidis had a remarkable aversion to low altitudes and exhibited maximum growth at 1,285 m. In conclusion, we speculate that one can predict what species will be found on railway sleepers and their probability and that species diversity primarily depends on sleeper type and altitude height. We believe that this study can contribute new insights into the ecology of black yeasts on railway sleepers and the railway factors that influence their diversity. [ABSTRACT FROM AUTHOR]

A total of fifty random samples of yoghurt collected from vendors and wholesale markets in El-Behera Governorate to assess their microbiological quality. The pH was determined and microbiological assessments were conducted in order to ascertain, the Coliforms count, Enterococci count, Staphylococcus aureus count and yeast count and mould count in the examined samples. Results given revealed that the mean values of pH was 4.06 ± 0.068, the mean value of Coliform count, Enterococci count, Staphylococcus aureus count, yeast count and mould counts were 5.6 x 104 ± 3.68 x 10³, 5.8 x 104 ± 5.43 x 10³, 5.5 x 104 ± 3.45 x 10³, 5.6 x 104 ± 4.9 x 10³ and 5.3 x 104 ± 3.33 x 10³ respectively, with respective incidences of 58, 28, 42, 28 and 40%. These findings suggest that yogurt traded in whole sale market in El-Behera Governorate has poor microbiological quality control. This poses danger to public health. Therefore, attention of the appropriate government agencies and manufacturers is needed to ensure that sale of yogurt by vendors is done in most appropriate condition and in a mobile refrigerator to maintain adequate temperature, thereby reduce contamination. [ABSTRACT FROM AUTHOR]

Sporulation, gametogenesis in yeast, consists of meiotic nuclear division and spore morphogenesis. In the fission yeast Schizosaccharomyces pombe, the four haploid nuclei produced after meiosis II are encapsulated by the forespore membrane (FSM), which is newly synthesized from spindle pole bodies (SPBs) in the cytoplasm of the mother cell as spore precursors. Although the coordination between meiosis and FSM assembly is vital for proper sporulation, the underlying mechanism remains unclear. In the present study, we identified a new meiosis-specific protein Npg1, and found that it was involved in the efficient formation of spores and spore viability. The accumulation and organization of the FSM was compromised in npgl-null cells, leading to the error-prone envelopment of nuclei. Npg1 was first seen as internuclear dots and translocated to the SPBs before the FSM assembled. Genetic analysis revealed that Npg1 worked in conjunction with the FSM proteins Spo3 and Meu14. These results suggest a possible signaling link from the nucleus to the meiotic SPBs in order to associate the onset of FSM assembly with meiosis II, which ensures the successful partitioning of gametic nuclei. [ABSTRACT FROM AUTHOR]

The rapid identification of yeast is essential for the optimization of antifungal therapy. The objective of our study was to evaluate two matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) platforms, the bioMérieux VITEK MS (IVD Knowledgebase v.2.0) and Bruker Biotyper (software version 3.1), for the rapid identification of medically relevant yeast. One hundred and seventeen isolates, representing six genera and 18 species, were analyzed using multiple direct smear methods to optimize identification. Sequence analysis was the gold standard for comparison. Isolates were analyzed with VITEK MS using the direct smear method +/− a 25 % formic acid on-plate extraction. For Biotyper, isolates were analyzed using direct smear without formic acid, and with 25 % and 100 % formic acid on-plate extractions. When all methods were included, VITEK MS correctly identified 113 (96.6 %) isolates after 24 h with one misidentification, and Biotyper correctly identified 77 (65.8 %) isolates using a threshold of ≥2.0 with no misidentifications. Using a revised threshold of ≥1.7, Biotyper correctly identified 103 (88.0 %) isolates, with 3 (2.6 %) misidentifications. For both platforms, the number of identifications was significantly increased using a formic acid overlay (VITEK MS, p < 0.01; Biotyper, p < 0.001), and reducing the Biotyper threshold from ≥2.0 to ≥1.7 significantly increased the rate of identification ( p < 0.001). The data in this study demonstrate that the direct smear method with on-plate formic acid extraction can be used for yeast identification on both MS platforms, and more isolates are identified using the VITEK MS system ( p < 0.01). [ABSTRACT FROM AUTHOR]

A universal feature of the response to stress and nutrient limitation is transcriptional upregulation of genes that encode proteins important for survival. Under many such conditions, the overall protein synthesis level is reduced, thereby dampening the stress response at the level of protein expression. For example, during glucose starvation in Saccharomyces cerevisiae (yeast), translation is rapidly repressed, yet the transcription of many stress- and glucose-repressed genes is increased. Here we show, using ribosomal profiling and microscopy, that this transcriptionally upregulated gene set consists of two classes: one class produces messenger RNAs that are translated during glucose starvation and are diffusely localized in the cytoplasm, including many heat-shock protein mRNAs; and the other class produces mRNAs that are not efficiently translated during glucose starvation and are concentrated in foci that co-localize with P bodies and stress granules, a class that is enriched for mRNAs involved in glucose metabolism. Surprisingly, the information specifying the differential localization and protein production of these two classes of mRNA is encoded in the promoter sequence: promoter responsiveness to heat-shock factor 1 (Hsf1) specifies diffuse cytoplasmic localization and higher protein production on glucose starvation. Thus, promoter sequences can influence not only the levels of mRNAs but also the subcellular localization of mRNAs and the efficiency with which they are translated, enabling cells to tailor protein production to the environmental conditions. [ABSTRACT FROM AUTHOR]

The objective of this study was to compare the nutritional composition and microbial quality of both branded and non-branded marmalades. Oranges were purchased from local markets while tangerines were product of in-house farm at Food Science and Product Development Institute (FSPDI), National Agricultural Research Centre (NARC), Islamabad. Juices were extracted and kept in sterilized and sealed glass bottles. Samples were subsequently analyzed for Total Plate Count (TPC), yeast, moulds, pH, sugars and vitamin C contents. Marmalade was prepared from blended juices of orange and tangerine. Branded marmalades i.e., National, Mitchells and Salman's were purchased from local markets and compared with non-branded (orange and tangerine) marmalades. Both marmalades were analysed for microbiological (TPC, yeast and mould), physico-chemical characteristics (pH, Brix, vitamin C and sugars) and organoleptic attributes. TPC count in both samples were nil. Yeast and mould count was nil in branded while it was 600 and 400 cfug-1 in orange and tangerine marmalades, respectively. Sugar contents, pH and Brix ranged from 14.97% to 59.70%, 3.00 to 3.46, 15.0 B to 60.2 B in branded and non-branded marmalades, respectively. Organoleptically marmalade developed at FSPDI was non-significantly different from all the branded marmalades except Salman's. [ABSTRACT FROM AUTHOR]

Yeast strain 25-G was isolated from naturally fermented cereal beverage (boza). By biochemical (API 20 C Aux) and molecular-genetic (partial sequencing of the 26S rRNA gene) methods, a representative of the species Saccharomyces cerevisiae var. diastaticus was identified. The enzymatic profile of the strain was determined by applying a kit system API ZYM (BioMerieux, France). Its proteolytic and amylase activities were examined as well. Saccharomyces cerevisiae var. diastaticus strain 25-G exhibits amylase activity, which makes it suitable for being included in the composition of starter cultures used at the production of fermented cereal foods and beverages. [ABSTRACT FROM AUTHOR]

Background and Objective: Biodeterioration is an irreversible damage that is caused by colonization of microorganisms on the surface of different materials. Among all microorganisms, fungi play an important role in deterioration of materials. Fungi can colonize on stone surfaces and by secreting different enzymes, organic and inorganic acids and pigments, can cause bio-weathering and changing not only the substrate materials but the color of stones. Furthermore, fungal mycelia can penetrate into the internal surfaces of stones and change the interior chemical contents of stones. Pasargadae including Cyrus the Great Tomb is entitled by UNESCO as one of the World Heritage Sites. This study was focused on the identification of fungi that were colonized on the tomb limestone monument. Materials and Methods: Sampling of stone was carried out to identify inhabiting molds and yeasts. biochemical and microscopic methods were used for isolated strains. In addition, the Polymerase Chain Reaction (PCR) and sequencing of the PCR products were done. Finally, phylogenic tree was constructed basde on the sequences of ITs region. Results and Conclusion: The common inhabiting fungi which isolated from the tomb limestone belong to Caldosporium sp., Embellisia sp., Cryptococcus sp., Candida sp., Meyerozyma sp., Arthirinium sp., Ulocladium sp., Fusarium sp., Humicola sp. and Pseudozyma sp., Stereomicroscopic and Scanning Electron Microscope images and XRD, were taken from pieces of stone samples and indicated the severe pattern damages such as pitting, biomineralization, etching and sugaring on the surfaces of stones. [ABSTRACT FROM AUTHOR]

Candida glabrata is one of the most common causes of candidemia, a life-threatening, systemic fungal infection, and is surpassed in frequency only by Candida albicans. Major factors contributing to the success of this opportunistic pathogen include its ability to readily acquire resistance to antifungals and to colonize and adapt to many different niches in the human body. Here we addressed the flexibility and adaptability of C. glabrata during interaction with macrophages with a serial passage approach. Continuous co-incubation of C. glabrata with a murine macrophage cell line for over six months resulted in a striking alteration in fungal morphology: The growth form changed from typical spherical yeasts to pseudohyphae-like structures – a phenotype which was stable over several generations without any selective pressure. Transmission electron microscopy and FACS analyses showed that the filamentous-like morphology was accompanied by changes in cell wall architecture. This altered growth form permitted faster escape from macrophages and increased damage of macrophages. In addition, the evolved strain (Evo) showed transiently increased virulence in a systemic mouse infection model, which correlated with increased organ-specific fungal burden and inflammatory response (TNFα and IL-6) in the brain. Similarly, the Evo mutant significantly increased TNFα production in the brain on day 2, which is mirrored in macrophages confronted with the Evo mutant, but not with the parental wild type. Whole genome sequencing of the Evo strain, genetic analyses, targeted gene disruption and a reverse microevolution experiment revealed a single nucleotide exchange in the chitin synthase-encoding CHS2 gene as the sole basis for this phenotypic alteration. A targeted CHS2 mutant with the same SNP showed similar phenotypes as the Evo strain under all experimental conditions tested. These results indicate that microevolutionary processes in host-simulative conditions can elicit adaptations of C. glabrata to distinct host niches and even lead to hypervirulent strains. [ABSTRACT FROM AUTHOR]

Antifreeze proteins (AFPs) are proteins with affinity towards ice and contribute to the survival of psychrophiles in subzero environment. Limited studies have been conducted on how AFPs from psychrophilic yeasts interact with ice. In this study, we describe the functional properties of an antifreeze protein from a psychrophilic Antarctic yeast, Glaciozyma antarctica. A cDNA encoding the antifreeze protein, AFP4, from G. antarctica PI12 was amplified from the mRNA extracted from cells grown at 4 °C. Sequence characterisation of Afp4 showed high similarity to fungal AFPs from Leucosporidium sp. AY30, LeIBP (93 %). The 786-bp cDNA encodes a 261-amino-acid protein with a theoretical pI of 4.4. Attempts to produce the recombinant Afp4 in Escherichia coli resulted in the formation of inclusion bodies (IB). The IB were subsequently denatured and refolded by dilution. Gel filtration confirmed that the refolded recombinant Afp4 is monomeric with molecular mass of ~25 kDa. Thermal hysteresis (TH) and recrystallisation inhibition assays confirmed the function of Afp4 as an antifreeze protein. In the presence of Afp4, ice crystals were modified into hexagonal shapes with TH values of 0.08 °C and smaller ice grains were observed compared with solutions without AFP. Structural analyses via homology modelling showed that Afp4 folds into β-helices with three distinct faces: a, b and c. Superimposition analyses predicted the b-face as the ice-binding surface of Afp4, whereby the mechanism of interaction is driven by hydrophobic interactions and the flatness of surface. This study may contribute towards an understanding of AFPs from psychrophilic yeasts. [ABSTRACT FROM AUTHOR]

Centrosomes--as well as the related spindle pole bodies (SPBs) of yeast--have been extensively studied from the perspective of their microtubule-organizing roles. Moreover, the biogenesis and duplication of these organelles have been the subject of much attention, and the importance of centrosomes and the centriole--ciliary apparatus for human disease is well recognized. Much less developed is our understanding of another facet of centrosomes and SPBs, namely their possible role as signalling centres. Yet, many signalling components, including kinases and phosphatases, have been associated with centrosomes and spindle poles, giving rise to the hypothesis that these organelles might serve as hubs for the integration and coordination of signalling pathways. In this review, we discuss a number of selected studies that bear on this notion. We cover different processes (cell cycle control, development, DNA damage response) and organisms (yeast, invertebrates and vertebrates), but have made no attempt to be comprehensive. This field is still young and although the concept of centrosomes and SPBs as signalling centres is attractive, it remains primarily a concept--in need of further scrutiny. We hope that this review will stimulate thought and experimentation. [ABSTRACT FROM AUTHOR]

Candida is the shortened name used to describe a class of fungi that includes more than 150 species of yeast. In healthy individuals, Candida exists harmlessly in mucus membranes such as your ears, eyes, gastrointestinal tract, mouth, nose, reproductive organs, sinuses, skin, stool and vagina, etc. It is known as your "beneficial flora" and has a useful purpose in the body. When an imbalance in the normal flora occurs, it causes an overgrowth of Candida albicans. The term is Candidiasis or Thrush. This is a fungal infection (Mycosis) of any of the Candida species, of which Candida albicans is the most common. When this happens, it can create a widespread havoc to our overall health and well-being of our body. [ABSTRACT FROM AUTHOR]

Invasive infections caused by yeasts are associated with high mortality and morbidity, and resistance to antifungal agents is increasing. Candida spp. has emerged as the leading cause of systemic nosocomial fungal infections. The aim of this study was to identify yeast isolates from sterile site specimens to species level, and to determine their susceptibility to fluconazole and voriconazole, at the National Health Laboratory, Service Dr George Mukhari Tertiary Laboratory from March to August 2007. Candida isolates were identified to species level using a germ tube test and/or Api® ID 32C kits. Antifungal susceptibility testing to fluconazole and voriconazole was performed using the disc diffusion method in accordance with the Clinical and Laboratory Standards Institute guidelines. All of the Candida isolates were from the neonatal intensive care unit (NICU), with the exception of two. The distribution of yeast isolates was as follows: C. krusei (41.9%), C. albicans (32.3%), C. inconspicua (5.5%), C. parapsilosis (2%), C. tropicalis (1.5%), C. sake (1.5%), C. lambica (1.5%), and C. valida (0.5%). Cryptococcus neoformans (11%), C. albidus (0.5%), Rhodotorula glutinis (1%), and C. humicola (0.5%) were also isolated. Of the isolated C. albicans, 61% were susceptible to fluconazole. A possible C. krusei outbreak could have occurred in the NICU during the study period. Voriconazole was the most susceptible antifungal agent to various yeast pathogens. The results of this study on azoles susceptibility testing of yeasts show that voriconazole may prove to be a valuable alternative antifungal agent in this tertiary hospital for the treatment of infections caused by yeasts, including Candida spp. [ABSTRACT FROM AUTHOR]