WorldWideScience

Sample records for cell factory aspergillus

  1. Aspergillus as a multi-purpose cell factory: current status and perspectives.

    Science.gov (United States)

    Meyer, Vera; Wu, Bo; Ram, Arthur F J

    2011-03-01

    Aspergilli have a long history in biotechnology as expression platforms for the production of food ingredients, pharmaceuticals and enzymes. The achievements made during the last years, however, have the potential to revolutionize Aspergillus biotechnology and to assure Aspergillus a dominant place among microbial cell factories. This mini-review will highlight most recent breakthroughs in fundamental and applied Aspergillus research with a focus on new molecular tools, techniques and products. New trends and concepts related to Aspergillus genomics and systems biology will be discussed as well as the challenges that have to be met to integrate omics data with metabolic engineering attempts.

  2. The Cell Factory Aspergillus Enters the Big Data Era: Opportunities and Challenges for Optimising Product Formation.

    Science.gov (United States)

    Meyer, Vera; Fiedler, Markus; Nitsche, Benjamin; King, Rudibert

    2015-01-01

    Living with limits. Getting more from less. Producing commodities and high-value products from renewable resources including waste. What is the driving force and quintessence of bioeconomy outlines the lifestyle and product portfolio of Aspergillus, a saprophytic genus, to which some of the top-performing microbial cell factories belong: Aspergillus niger, Aspergillus oryzae and Aspergillus terreus. What makes them so interesting for exploitation in biotechnology and how can they help us to address key challenges of the twenty-first century? How can these strains become trimmed for better growth on second-generation feedstocks and how can we enlarge their product portfolio by genetic and metabolic engineering to get more from less? On the other hand, what makes it so challenging to deduce biological meaning from the wealth of Aspergillus -omics data? And which hurdles hinder us to model and engineer industrial strains for higher productivity and better rheological performance under industrial cultivation conditions? In this review, we will address these issues by highlighting most recent findings from the Aspergillus research with a focus on fungal growth, physiology, morphology and product formation. Indeed, the last years brought us many surprising insights into model and industrial strains. They clearly told us that similar is not the same: there are different ways to make a hypha, there are more protein secretion routes than anticipated and there are different molecular and physical mechanisms which control polar growth and the development of hyphal networks. We will discuss new conceptual frameworks derived from these insights and the future scientific advances necessary to create value from Aspergillus Big Data.

  3. Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88

    DEFF Research Database (Denmark)

    Pel, Herman J.; de Winde, Johannes H.; Archer, David B.

    2007-01-01

    The filamentous fungus Aspergillus niger is widely exploited by the fermentation industry for the production of enzymes and organic acids, particularly citric acid. We sequenced the 33.9-megabase genome of A. niger CBS 513.88, the ancestor of currently used enzyme production strains. A high level...

  4. 曲霉菌细胞工厂的现状及前景%Aspergillus as a potential cell factory:current status and perspectives

    Institute of Scientific and Technical Information of China (English)

    顾丰颖; 高洁; 何国庆

    2012-01-01

    曲霉菌是一类重要的工业微生物,在传统发酵工业中是多种食品成分、酶制剂及有机酸等的主要生产者。随着技术的发展,曲霉菌的开发利用得到了更广泛的关注,亦逐步被开发成为多表达的细胞工厂、本文从新技术新产品等方面对曲霉作为细胞工厂近年来在基础研究及应用上的进展及突破做一概述。对以曲霉菌为表达平台而进行的基因组学、代谢组学、蛋白组学等组学研究予以综述,并对以曲霉菌为代表的细胞工厂的发展前景及趋势提出新的构思。%The genus Aspergilli is an important industrial workhorse with extensive application in variety of fermentation industrial process, like producing food ingredient, enzymic preparations and organic acid etc.With the development of the technologies, the exploitation of Aspergilli has attracted more attention and gradually been developed to be the multi-purpose of cell factories. The review will highlight most recent breakthroughs in fundamental and applied Aspergillus research with a focus on new molecular tools and products. Genomics, transcriptomics, proteomics and metabolomics of Aspergilli has been summarized. New trends and concepts related to Aspergillus as the cell factory will be discussed.

  5. Engineering the Polyketide Cell Factory

    DEFF Research Database (Denmark)

    Mølgaard, Louise

    sufficient titers. To improve the production of polyketides biological engineering principles have been applied for the development and engineering of microbial polyketide cell factories. The two biological hosts used for heterologous polyketide production were Aspergillus nidulans and Saccharomyces...... phosphopantetheinylase (PPTase). This versatile vector system can easily be used for expression of other polyketides of interest as well as extended to express whole gene clusters. After achieving proof of principle in terms of expression, the polyketide cell factory must be optimized. The optimization can be achieved...... characterization in bioreactors revealed that the yields of 6-MSA on biomass increased albeit not significantly. As a result of this it may be argued that there is still more work to be done in terms of model building in A. nidulans. Utilizing another well-established cell factory S. cerevisiae the capabilities...

  6. Cell Factory Engineering.

    Science.gov (United States)

    Davy, Anne Mathilde; Kildegaard, Helene Faustrup; Andersen, Mikael Rørdam

    2017-03-22

    Rational approaches to modifying cells to make molecules of interest are of substantial economic and scientific interest. Most of these efforts aim at the production of native metabolites, expression of heterologous biosynthetic pathways, or protein expression. Reviews of these topics have largely focused on individual strategies or cell types, but collectively they fall under the broad umbrella of a growing field known as cell factory engineering. Here we condense >130 reviews and key studies in the art into a meta-review of cell factory engineering. We identified 33 generic strategies in the field, all applicable to multiple types of cells and products, and proven successful in multiple major cell types. These apply to three major categories: production of native metabolites and/or bioactives, heterologous expression of biosynthetic pathways, and protein expression. This meta-review provides general strategy guides for the broad range of applications of rational engineering of cell factories.

  7. Colorful Microbial Cell Factories

    DEFF Research Database (Denmark)

    Petersen, Pia Damm

    Yeast cell factories are powerful tools used for the production of high-value natural compounds otherwise not easily available. Many bioactive and industrially important plant secondary metabolites can be produced in yeast by engineering their biosynthetic pathways into yeast cells, as these both...... anthocyanins. Yeast cell factories present a platform to circumvent the problem of low yields of interesting molecular structures in plant tissues, as hand-picking of desired enzyme activities allows for specific biosynthesis of the precise pigment of interest, as well as choosing more stable structures...... for heterologous biosynthesis is possible. In cell factories, great improvements in yields can be achieved through molecular engineering of flux from endogenous yeast precursors, e.g. by elimination of by-product formation, and by genetic optimization of pathway components, such as fine-tuning of expression levels...

  8. Cell Factory Engineering

    DEFF Research Database (Denmark)

    Davy, Anne Mathilde; Kildegaard, Helene Faustrup; Andersen, Mikael Rørdam

    2017-01-01

    Rational approaches to modifying cells to make molecules of interest are of substantial economic and scientific interest. Most of these efforts aim at the production of native metabolites, expression of heterologous biosynthetic pathways, or protein expression. Reviews of these topics have largely...... focused on individual strategies or cell types, but collectively they fall under the broad umbrella of a growing field known as cell factory engineering. Here we condense >130 reviews and key studies in the art into a meta-review of cell factory engineering. We identified 33 generic strategies...... in the field, all applicable to multiple types of cells and products, and proven successful in multiple major cell types. These apply to three major categories: production of native metabolites and/or bioactives, heterologous expression of biosynthetic pathways, and protein expression. This meta...

  9. Improved premises for cell factory development

    DEFF Research Database (Denmark)

    Søgaard, Karina Marie

    is at the core of biotechnology and numerous molecular tools and bacterial strains have been developed over the past four decades for this purpose. Understanding of the genetic code and our ability to manipulate genetic material, paves the way for the microbial cell factory development that enables production...... of protein in a sustainable, costefficient manner. In this thesis I report the joined efforts of my colleagues and myself, to improve the premises for cell factory development by optimizing the cloning strategies, improving the awareness of unforeseen side-effects in complex bacterial expression systems...... have improved the premises for cell factory development in the future....

  10. Cell factories for a sustainable world

    DEFF Research Database (Denmark)

    Kindtler, Jens William

    2016-01-01

    The Novo Nordisk Foundation Center for Biosustainability (CFB) is a university research center under the Technical University of Denmark within the field of design, engineering and construction of superior cell factories for the production of biopharmaceuticals, anti-infective compounds, industrial...

  11. Targeting population heterogeneity for optimal cell factories

    DEFF Research Database (Denmark)

    Heins, Anna-Lena; Carlqvist, Magnus; Helmark, S.

    analysis, and thereby created the possibility to map population heterogeneity. A factorial design with pH, glucose concentration and oxygen level was performed in batch cultivations using the growth reporter strains to evaluate the effect of those environmental factors on heterogeneity level and amount...... of living cells. A highly dynamic behavior with regard to subpopulation distribution during the different growth stages was seen for the batch cultivations. Moreover, it could be demonstrated that the glucose concentration had a clear influence on the heterogeneity. The results from the factorial design...

  12. Cell Factory Stability and Genetic Circuits for Improved Strain Development

    DEFF Research Database (Denmark)

    Rugbjerg, Peter

    Development of new chemical-­‐producing microbial cell factories is an iterative trial-­and-­error process, and to screen candidate cells at high throughput, genetic biosensor systems are appealing. Each biosensor has distinct biological parameters, making modular tuning networks attractive....... However, all synthetic gene systems -­ including the target metabolic pathways themselves -­ represent a possible fitness burden to the cell and thus constitute a threat to strain stability. In this thesis, several studies served to develop genetic systems for optimizing cell factory development...... factories in future....

  13. Kinetic models in industrial biotechnology - Improving cell factory performance.

    Science.gov (United States)

    Almquist, Joachim; Cvijovic, Marija; Hatzimanikatis, Vassily; Nielsen, Jens; Jirstrand, Mats

    2014-07-01

    An increasing number of industrial bioprocesses capitalize on living cells by using them as cell factories that convert sugars into chemicals. These processes range from the production of bulk chemicals in yeasts and bacteria to the synthesis of therapeutic proteins in mammalian cell lines. One of the tools in the continuous search for improved performance of such production systems is the development and application of mathematical models. To be of value for industrial biotechnology, mathematical models should be able to assist in the rational design of cell factory properties or in the production processes in which they are utilized. Kinetic models are particularly suitable towards this end because they are capable of representing the complex biochemistry of cells in a more complete way compared to most other types of models. They can, at least in principle, be used to in detail understand, predict, and evaluate the effects of adding, removing, or modifying molecular components of a cell factory and for supporting the design of the bioreactor or fermentation process. However, several challenges still remain before kinetic modeling will reach the degree of maturity required for routine application in industry. Here we review the current status of kinetic cell factory modeling. Emphasis is on modeling methodology concepts, including model network structure, kinetic rate expressions, parameter estimation, optimization methods, identifiability analysis, model reduction, and model validation, but several applications of kinetic models for the improvement of cell factories are also discussed.

  14. Novel and improved yeast cell factories for biosustainable processes

    DEFF Research Database (Denmark)

    Workman, Mhairi

    2014-01-01

    utilizing traditionally applied cell factories are generally based on a limited range of substrates (mainly glucose). However, a wider diversity in substrate range is highly desirable in developing biorefinery scenarios where feed-stocks containing a number of carbon sources are typically employed....... In addition to plant biomass hydrolysates, glycerol is of interest here, being available in amounts relevant for industrial scale bioprocesses due to increased production of biodiesel. The well characterised cell factory Saccharomyces cerevisiae exhibits a clear preference for glucose as a carbon source...... with relevant applications as cell factories (including Pichia spp. and Yarrowia lipolytica) and other less well characterized strains (e.g. Pachysolen tannophilus). This presentation will address how we evaluate cellular performance with a view to utilizing yeast species in industrial biotechnology...

  15. Characterizing Yarrowia lipolytica as a cell factory for sustainable bioprocesses

    DEFF Research Database (Denmark)

    Lubuta, Patrice; Workman, Mhairi

    With current global focus on renewable and sustainable technologies, there is a strong need to develop innovative solutions in industrial biotechnology. The non-conventional yeast Yarrowia lipolytica has considerable potential as a versatile cell factory, especially when compared to the typically...

  16. Metabolic shifts: a fitness perspective for microbial cell factories

    NARCIS (Netherlands)

    Goel, A.; Wortel, M.T.; Molenaar, D.; Teusink, B.

    2012-01-01

    Performance of industrial microorganisms as cell factories is limited by the capacity to channel nutrients to desired products, of which optimal production usually requires careful manipulation of process conditions, or strain improvement. The focus in process improvement is often on understanding a

  17. A versatile expression vector system for mammalian cell factories

    DEFF Research Database (Denmark)

    Lund, Anne Mathilde; Kildegaard, Helene Faustrup; Hansen, Bjarne Gram

    The development of the field of mammalian cell factories requests fast and high-throughput methods which means high need for simpler and more efficient cloning techniques. This project applies the ligation-free USERTM (uracil-specific excision reagent) cloning technique to construct mammalian...

  18. Synthetic biology of cyanobacterial cell factories

    NARCIS (Netherlands)

    Angermayr, S.A.

    2014-01-01

    In the field of microbial biotechnology rational design approaches are employed for the generation of microbial cells with desired functions, such as the ability to produce precursor molecules for biofuels or bioplastics. In essence, that is the introduction of a (new) biosynthetic pathway into a mi

  19. Chitosan Obtained from Cell Wall of Aspergillus Niger Mycelium

    Institute of Scientific and Technical Information of China (English)

    HUANG Hui-li; LIN Wen-luan; LIN Jian-ming

    2004-01-01

    Chitin from cell walls of Aspergillus Niger mycelium was prepared. A new method for the preparation of high deacetylation degree chitosan was studied in a dilute sodium hydroxide solution at a high pressure. The experimental results indicate that the deacetylation degree of the chitosan can reach 80% under the condition of a 5.00 mol/L sodium hydroxide solution at 0.1 MPa of pressure for 1 h. This method shows the advantages of the applications in the industry production and environment protection.

  20. Establishing new microbial cell factories for sustainable bioprocesses

    DEFF Research Database (Denmark)

    Workman, Mhairi; Holt, Philippe; Liu, Xiaoying

    2012-01-01

    and developed. Systems biology approaches can be applied with these well studied strains, where considerable physiological data has been obtained, genome sequences are known and genetic tools have been developed. With the necessity of considering a wider array of substrates and the need for new types...... of products, it may be interesting to look to less domesticated strains and towards more non-conventional hosts in the development of new bioprocesses. This approach demands thorough physiological characterization as well as establishment of tools for genetic engineering if new cell factories...

  1. Morphogenesis and protein production in Aspergillus niger

    NARCIS (Netherlands)

    Kwon, Min Jin

    2014-01-01

    The research described in this thesis aims to get more fundamental insights in the molecular mechanisms used by Aspergillus niger in relation to control morphology and protein secretion. Knowledge on these two aspects is highly relevant to further optimization of A.niger as a cell factory

  2. BIOSORPTION OF CR (VI BY RESTING CELLS OF ASPERGILLUS SP.

    Directory of Open Access Journals (Sweden)

    M. Sen , M. Ghosh Dastidar

    2007-01-01

    Full Text Available Biosorption of Cr(VI from aqueous solution was studied in a batch bioreactor using the resting cells of filamentous fungal biomass (Aspergillus sp. isolated from industrial wastewaters. The specific Cr(VI removal (mg/g of dried biomass decreased with increase in pH and increased with increase in initial Cr(VI concentration, upto 500 mg/L. By increasing biomass concentration from 2.4 to 5.2 g/L, the specific metal removal remained almost constant. The studies carried out by using the resting cells from various stages of growth indicated maximum Cr(VI removal of 34.8 mg/g using the biomass from the beginning of the stationary phase. The adsorption equilibrium constants Qº (42.9 mg/g and b (0.0091/mg were obtained from the Langmuir adsorption isotherm model.

  3. Synthetic yeast based cell factories for vanillin-glucoside production

    DEFF Research Database (Denmark)

    Strucko, Tomas

    allowing for the introduction of large and complex metabolic pathways need to be added to the existing repertoire. To reduce the number of gene engineering steps required for cell factory construction, a new set of integrative “EasyClone” vectors have been developed in this study. This platform enables...... simultaneous integration of multiple genes with an option of recycling selection markers. Moreover, EasyClone vectors combine the advantage of efficient uracil-excision reaction based cloning that allows integration of one or two genes per plasmid and Cre-LoxP mediated marker recycling system. As a proof...... of concept, it was demonstrated that using EasyClone system it is possible to simultaneously integrate three DNA fragments carrying genes encoding for either yellow, cyan or red fluorescent proteins. In addition, all genetic markers were successfully removed using Cre-mediated recombination without...

  4. Cell biology of the Koji mold Aspergillus oryzae.

    Science.gov (United States)

    Kitamoto, Katsuhiko

    2015-01-01

    Koji mold, Aspergillus oryzae, has been used for the production of sake, miso, and soy sauce for more than one thousand years in Japan. Due to the importance, A. oryzae has been designated as the national micro-organism of Japan (Koku-kin). A. oryzae has been intensively studied in the past century, with most investigations focusing on breeding techniques and developing methods for Koji making for sake brewing. However, the understanding of fundamental biology of A. oryzae remains relatively limited compared with the yeast Saccharomyces cerevisiae. Therefore, we have focused on studying the cell biology including live cell imaging of organelles, protein vesicular trafficking, autophagy, and Woronin body functions using the available genomic information. In this review, I describe essential findings of cell biology of A. oryzae obtained in our study for a quarter of century. Understanding of the basic biology will be critical for not its biotechnological application, but also for an understanding of the fundamental biology of other filamentous fungi.

  5. Decolorization of reactive Brilliant Blue KN-R by immobilized cells of Aspergillus ficuum

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Aspergillus ficuum was immobilized with sodium alginate, and decolorization of Reactive Brilliant Blue KN-R was studied on immobilized and free Aspergillus ficuum. The optimal preparation condition of the strain immobilization was obtained by the orthogonal test, it is sodium alginate 3%, CaCl2 5%, wet mycelia 30 g/L, calcific time 8 h. It was found that the immobilized cells could effectively decolorize Reactive Brilliant Blue KN-R, the optimum temperature and pH were 33℃ and 5.0, respectively. The kinetics study of decolorization of immobilized cells showed that the decolorization of Aspergillus ficuum immobilized conformed to zero-order reaction model. The decolorization efficiency of immobilized cell compared with that of free cell in different physical conditions. Results showed that the decolorization of immobilized cells with mycelia had the best efficiency. The immobilized cells could be reused after the first decolorization.

  6. Expression in Escherichia coli, refolding and crystallization of Aspergillus niger feruloyl esterase A using a serial factorial approach

    NARCIS (Netherlands)

    Benoit, Isabelle; Coutard, Bruno; Oubelaid, Rachid; Asther, Marcel; Bignon, Christophe

    2007-01-01

    Hydrolysis of plant biomass is achieved by the combined action of enzymes secreted by microorganisms and directed against the backbone and the side chains of plant cell wall polysaccharides. Among side chains degrading enzymes, the feruloyl esterase A (FAEA) specifically removes feruloyl residues. T

  7. METABOLIC MODELLING IN THE DEVELOPMENT OF CELL FACTORIES BY SYNTHETIC BIOLOGY

    Directory of Open Access Journals (Sweden)

    Paula Jouhten

    2012-10-01

    Full Text Available Cell factories are commonly microbial organisms utilized for bioconversion of renewable resources to bulk or high value chemicals. Introduction of novel production pathways in chassis strains is the core of the development of cell factories by synthetic biology. Synthetic biology aims to create novel biological functions and systems not found in nature by combining biology with engineering. The workflow of the development of novel cell factories with synthetic biology is ideally linear which will be attainable with the quantitative engineering approach, high-quality predictive models, and libraries of well-characterized parts. Different types of metabolic models, mathematical representations of metabolism and its components, enzymes and metabolites, are useful in particular phases of the synthetic biology workflow. In this minireview, the role of metabolic modelling in synthetic biology will be discussed with a review of current status of compatible methods and models for the in silico design and quantitative evaluation of a cell factory.

  8. Industrial Systems Biology of Saccharomyces cerevisiae Enables Novel Succinic Acid Cell Factory

    DEFF Research Database (Denmark)

    Otero, José Manuel; Cimini, Donatella; Patil, Kiran Raosaheb

    2013-01-01

    Saccharomyces cerevisiae is the most well characterized eukaryote, the preferred microbial cell factory for the largest industrial biotechnology product (bioethanol), and a robust commerically compatible scaffold to be exploitted for diverse chemical production. Succinic acid is a highly sought......-direction of carbon fluxes in S. cerevisiae, and hence show proof of concept that this is a potentially attractive cell factory for over-producing different platform chemicals....

  9. Interaction of Aspergillus with alveolar Type II cells and phagocytes

    NARCIS (Netherlands)

    Escobar Salazar, N.

    2016-01-01

    Aspergillus species are worldwide distributed fungi and abundant in nature. Aspergilli are mainly saprotrophic obtaining nutrients by degrading dead organic material in particular that of plants. Currently, around 837 species have been reported. Due to the broad range of compound secreted by Aspergi

  10. ENHANCED PRODUCTION OF PECTINOLYTIC ENZYMES FROM IMMOBILIZED CELLS OF MIXED ASPERGILLUS SPECIES

    Directory of Open Access Journals (Sweden)

    Shruti Singh

    2012-12-01

    Full Text Available The cells of isolated mixed culture of Aspergillus fumigatus and Aspergillus sydowii were immobilized in calcium alginate beads. Studies were carried out on different parameters like alginate concentration, incubation time and bead inoculum which affects the productivity and stability of the immobilized system. The best enzymatic activities were obtained with 3% alginate concentration, 48h of incubation time and 200 beads/flask of inoculum. Optimization of these factors causes an increase in enzymatic activities and the possibility of semicontinuous cultivation. Immobilized cells could be reused in five successive reaction cycles with a slight decrease in activities.

  11. [Immobilization of mixed bacteria by microcapsulation for hydrogen production--a trial of pseudo "Cell Factory"].

    Science.gov (United States)

    Ma, Qianlan; Lin, Dongqiang; Yao, Shanjing

    2010-10-01

    Sodium cellulose sulfate (NaCS)/Ploy-dimethyl-dially-ammonium-chloride (PDMDAAC) microcapsules were used as a novel pseudo "Cell Factory" to immobilize mixed bacteria for hydrogen production under anaerobic conditions. Compared to free cells, the hydrogen production was increased more than 30% with NaCS/PDMDAAC microcapsules as the pseudo "Cell Factory". The biomass was increased from 1.5 g/L in free cell culture to 3.2 g/L in the pseudo "Cell Factory". This pseudo "Cell Factory" system showed the excellent stability during 15 repeated-batches. The hydrogen yield maintained 1.73-1.81 mol H2/mol glucose. The fermentation cycle was shortened from 48 h to 24 h, resulting in an increase of 198.6% in the hydrogen production rate. There were high percentage of butyric acid and acetic acid in the culture broth, which meant that the pseudo "Cell Factory" established in the present work could be used for the multi-product system.

  12. Aspergillus galactomannan antigen levels in allogeneic haematopoietic stem cell transplant recipients given total parenteral nutrition.

    NARCIS (Netherlands)

    Blijlevens, N.M.A.; Donnelly, J.P.; Meis, J.F.G.M.; Verweij, P.E.; Pauw, B.E. de

    2002-01-01

    False-positive tests for Aspergillus galactomannan have been reported in neutropenic patients. We failed to detect any circulating antigen during the 2 weeks following allogeneic haematopoietic stem cell transplantation of 12 patients who had severe mucositis but were unable to eat.

  13. Biocatalytic Resolution of para-Nitrostyrene Oxide by Resting Cells of Different Aspergillus niger Strains

    Institute of Scientific and Technical Information of China (English)

    金浩; 李祖义; 王清

    2001-01-01

    Biocatalytic resolution of racemic para-nitrostyrene oxide was accomplished by employing the epoxide hydrolases from the whole cells of several Aspergillus niger (A. niger) strains. In the cases investigated, excellent selectivity was achieved with such strains as A, niger 5450, A. niger 5320.

  14. Evaluation of cell factory performance through determination of intracellular metabolites using LC-MS/MS

    DEFF Research Database (Denmark)

    Magdenoska, Olivera; Martinussen, Jan; Nielsen, Kristian Fog

    2012-01-01

    A major objective in biotechnology is the improvement of the efficiency of host microorganisms used as cell factories. Engineering a strain capable of producing high amounts of a desired biochemical is a multi-step process consisting of design, construction, and analysis of the constructed cell...... factory. In order to address the function or disfunction of the engineered cells,systems biology tools are employed by using the multi “omics” approach (genomics, transcriptomics, proteomics, metabolomics and fluxomics). Metabolomics is a tool aimed at a quantitative understanding of metabolism...... analysis, (iii) data analysis and interpretation. The established analytical method covers analysis of sixty metabolites from glycolysis,cofactors, coenzymes and nucleotides. Implementation of this method provides a powerful new tool in future cell factory design and characterization....

  15. Industrial systems biology and its impact on synthetic biology of yeast cell factories.

    Science.gov (United States)

    Fletcher, Eugene; Krivoruchko, Anastasia; Nielsen, Jens

    2016-06-01

    Engineering industrial cell factories to effectively yield a desired product while dealing with industrially relevant stresses is usually the most challenging step in the development of industrial production of chemicals using microbial fermentation processes. Using synthetic biology tools, microbial cell factories such as Saccharomyces cerevisiae can be engineered to express synthetic pathways for the production of fuels, biopharmaceuticals, fragrances, and food flavors. However, directing fluxes through these synthetic pathways towards the desired product can be demanding due to complex regulation or poor gene expression. Systems biology, which applies computational tools and mathematical modeling to understand complex biological networks, can be used to guide synthetic biology design. Here, we present our perspective on how systems biology can impact synthetic biology towards the goal of developing improved yeast cell factories. Biotechnol. Bioeng. 2016;113: 1164-1170. © 2015 Wiley Periodicals, Inc.

  16. A second-generation Bacillus cell factory for rare inositol production.

    Science.gov (United States)

    Tanaka, Kosei; Takanaka, Shinji; Yoshida, Ken-ichi

    2014-01-01

    Some rare inositol stereoisomers are known to exert specific health-promoting effects, including scyllo-inositol (SI), which is a promising therapeutic agent for Alzheimer disease. We recently reported a Bacillus subtilis cell factory that performed the efficient production of SI from the cheapest and most abundant isomer myo-inositol (MI). In the cell factory all "useless" genes involved in MI and SI metabolism were deleted and overexpression of the key enzymes, IolG and IolW, was appended. It converted 10 g/L MI into the same amount of SI in 48 h of cultivation. In this addendum, we discuss further improvement in the cell factory and its possible applications.

  17. A second-generation Bacillus cell factory for rare inositol production

    Science.gov (United States)

    Tanaka, Kosei; Takanaka, Shinji; Yoshida, Ken-ichi

    2014-01-01

    Some rare inositol stereoisomers are known to exert specific health-promoting effects, including scyllo-inositol (SI), which is a promising therapeutic agent for Alzheimer disease. We recently reported a Bacillus subtilis cell factory that performed the efficient production of SI from the cheapest and most abundant isomer myo-inositol (MI). In the cell factory all “useless” genes involved in MI and SI metabolism were deleted and overexpression of the key enzymes, IolG and IolW, was appended. It converted 10 g/L MI into the same amount of SI in 48 h of cultivation. In this addendum, we discuss further improvement in the cell factory and its possible applications. PMID:25482235

  18. Transcription factors as targets for improving Aspergillus niger as cell factory

    DEFF Research Database (Denmark)

    Poulsen, Lars; Bruno, K.S.; Thykær, Jette

    Altering fluxes for overcoming metabolic bottlenecks have traditionally been approached by genetic engineering of a single or few metabolic genes. This strategy struggles to overcome the subjacent regulation thus the outcome has frequently shown to be of limited success. Transcription factors hav...

  19. Droplet size influences division of mammalian cell factories in droplet microfluidic cultivation

    DEFF Research Database (Denmark)

    Periyannan Rajeswari, Prem Kumar; Joensson, Haakan N.; Svahn, Helene Andersson

    2017-01-01

    The potential of using droplet microfluidics for screening mammalian cell factories has been limited by the difficulty in achieving continuous cell division during cultivation in droplets. Here, we report the influence of droplet size on mammalian cell division and viability during cultivation......, making them a more suitable droplet size for 72-h cultivation. This study shows a direct correlation of microfluidic droplet size to the division and viability of mammalian cells. This highlights the importance of selecting suitable droplet size for mammalian cell factory screening assays....... in droplets. Chinese Hamster Ovary (CHO) cells, the most widely used mammalian host cells for biopharmaceuticals production were encapsulated and cultivated in 33, 180 and 320 pL droplets for 3 days. Periodic monitoring of the droplets during incubation showed that the cell divisions in 33 pL droplets stopped...

  20. Production of Useful Terpenoids by Higher-Fungus Cell Factory and Synthetic Biology Approaches.

    Science.gov (United States)

    Xiao, Han; Zhong, Jian-Jiang

    2016-03-01

    Higher fungi with greater than 70000 species are regarded as a rich source of various natural compounds including terpenoids, the production of which represents a wide range of interest in pharmaceutical and healthcare industries. This review summarizes the current knowledge of terpenoids synthesized by higher fungi, and highlights the current state-of-the-art regarding genetic manipulation of higher fungi. As the focus, this article will discuss the most recent approaches enabling native hosts and heterologous microbes to efficiently produce various terpenoids, especially with regard to the construction of 'smart' higher-fungus cell factories. The merits and demerits of heterologous versus native hosts as cell factories will also be debated.

  1. Development of a yeast cell factory for production of aromatic secondary metabolites

    DEFF Research Database (Denmark)

    Rodriguez Prado, Edith Angelica

    secondary metabolites in cell factories. In this research project, we developed a yeast platform strain for the production of p-coumaric acid an intermediate compound for the synthesis of aromatic secondary metabolites. Subsequently, we performed a systems biology analysis of the strain and finally we...... developed an array of yeast strains expressing flavonoid metabolic pathways containing up to ten heterologous genes. The platform strain was capable of producing 1.93 ± 0.26 g L-1 of p-coumaric acid in fed-batch fermentation, which is the highest titer that has been reported for a yeast cell factory so far...

  2. GDP-mannose pyrophosphorylase is essential for cell wall integrity, morphogenesis and viability of Aspergillus fumigatus.

    Science.gov (United States)

    Jiang, Hechun; Ouyang, Haomiao; Zhou, Hui; Jin, Cheng

    2008-09-01

    GDP-mannose pyrophosphorylase (GMPP) catalyses the synthesis of GDP-mannose, which is the precursor for the mannose residues in glycoconjugates, using mannose 1-phosphate and GTP as substrates. Repression of GMPP in yeast leads to phenotypes including cell lysis, defective cell wall, and failure of polarized growth and cell separation. Although several GMPPs have been isolated and characterized in filamentous fungi, the physiological consequences of their actions are not clear. In this study, Afsrb1, which is a homologue of yeast SRB1/PSA1/VIG9, was identified in the Aspergillus fumigatus genome. The Afsrb1 gene was expressed in Escherichia coli, and recombinant AfSrb1 was functionally confirmed as a GMPP. By the replacement of the native Afsrb1 promoter with an inducible Aspergillus nidulans alcA promoter, the conditional inactivation mutant strain YJ-gmpp was constructed. The presence of 3 % glucose completely blocked transcription of P(alcA)-Afsrb1, and was lethal to strain YJ-gmpp. Repression of Afsrb1 expression in strain YJ-gmpp led to phenotypes including hyphal lysis, defective cell wall, impaired polarity maintenance, and branching site selection. Also, rapid germination and reduced conidiation were documented. However, in contrast to yeast, strain YJ-gmpp retained the ability to direct polarity establishment and septation. Our results showed that the Afsrb1 gene is essential for cell wall integrity, morphogenesis and viability of Aspergillus fumigatus.

  3. Development of advanced therapies in Italy: Management models and sustainability in six Italian cell factories.

    Science.gov (United States)

    Gaipa, Giuseppe; Introna, Martino; Golay, Josee; Nolli, Maria Luisa; Vallanti, Giuliana; Parati, Eugenio; Giordano, Rosaria; Romagnoli, Luca; Melazzini, Mario; Biondi, Andrea; Biagi, Ettore

    2016-04-01

    On November 10, 2014, the representatives of all six certified Good Manufacturing Practices (GMP) cell factories operating in the Lombardy Region of Italy convened a 1-day workshop in Milan titled "Management Models for the Development And Sustainability of Cell Factories: Public-Private Partnership?" The speakers and panelists addressed not only the many scientific, technological and cultural challenges faced by Lombardy Cell Factories, but also the potential impact of advanced therapy medicinal products (ATMPs) on public health and the role played by translational research in this process. Future perspectives for research and development (R&D) and manufacturing processes in the field of regenerative medicine were discussed as well. This report summarizes the most important issues raised by the workshop participants with particular emphasis on strengths and limitations of the R&D and manufacturing processes for innovative therapeutics in Lombardy and what can be improved in this context while maintaining GMP standards. The participants highlighted several strategies to translate patient-specific advanced therapeutics into scaled manufacturing products for clinical application. These included (i) the development of a synergistic interaction between public and private institutions, (ii) better integration with Italian regulatory agencies and (iii) the creation of a network among Lombardy cell factories and other Italian and European institutions.

  4. The Uses and Future Prospects of Metabolomics and Targeted Metabolite Profiling in Cell Factory Development

    DEFF Research Database (Denmark)

    Harrison, Scott James; Herrgard, Markus

    2013-01-01

    The development of cell factories for the production of chemicals has traditionally relied on measurements of product metabolite titers to assess the performance of genetically manipulated strains. With the development of improved metabolomics and targeted metabolite profiling methods, these broa......The development of cell factories for the production of chemicals has traditionally relied on measurements of product metabolite titers to assess the performance of genetically manipulated strains. With the development of improved metabolomics and targeted metabolite profiling methods......, these broader measurements of the cellular metabolic state are now becoming part of the toolbox used to characterize cell factories. In this review we briefly summarize the benefits and challenges of global metabolomics and targeted metabolite profiling methods and discuss the application of these methods...... in both pathway discovery and cell factory engineering. We focus particularly on exploring the potential of global metabolomics to complement more traditional targeted methods. We conclude the review by discussing emerging trends in metabolomics and how these developments can aid the engineering of better...

  5. Development of a yeast cell factory for production of aromatic products

    DEFF Research Database (Denmark)

    Rodriguez Prado, Edith Angelica; Kildegaard, Kanchana Rueksomtawin; Li, Mingji

    2014-01-01

    There is much interest in aromatic chemicals in the chemical industry as these can be used for production of dyes, anti-oxidants, nutraceuticals and food ingredients. Yeast is a widely used cell factory and it is particularly well suited for production of aromatic chemicals via complex biosynthetic...

  6. On the Use of Metabolic Control Analysis in the Optimization of Cyanobacterial Biosolar Cell Factories.

    NARCIS (Netherlands)

    Angermayr, S.; Hellingwerf, K.J.

    2013-01-01

    Oxygenic photosynthesis will have a key role in a sustainable future. It is therefore significant that this process can be engineered in organisms such as cyanobacteria to construct cell factories that catalyze the (sun)light-driven conversion of CO2 and water into products like ethanol, butanol, or

  7. Production of fatty acid-derived oleochemicals and biofuels by synthetic yeast cell factories.

    Science.gov (United States)

    Zhou, Yongjin J; Buijs, Nicolaas A; Zhu, Zhiwei; Qin, Jiufu; Siewers, Verena; Nielsen, Jens

    2016-01-01

    Sustainable production of oleochemicals requires establishment of cell factory platform strains. The yeast Saccharomyces cerevisiae is an attractive cell factory as new strains can be rapidly implemented into existing infrastructures such as bioethanol production plants. Here we show high-level production of free fatty acids (FFAs) in a yeast cell factory, and the production of alkanes and fatty alcohols from its descendants. The engineered strain produces up to 10.4 g l(-1) of FFAs, which is the highest reported titre to date. Furthermore, through screening of specific pathway enzymes, endogenous alcohol dehydrogenases and aldehyde reductases, we reconstruct efficient pathways for conversion of fatty acids to alkanes (0.8 mg l(-1)) and fatty alcohols (1.5 g l(-1)), to our knowledge the highest titres reported in S. cerevisiae. This should facilitate the construction of yeast cell factories for production of fatty acids derived products and even aldehyde-derived chemicals of high value.

  8. Materials for stem cell factories of the future

    Science.gov (United States)

    Celiz, Adam D.; Smith, James G. W.; Langer, Robert; Anderson, Daniel G.; Winkler, David A.; Barrett, David A.; Davies, Martyn C.; Young, Lorraine E.; Denning, Chris; Alexander, Morgan R.

    2014-06-01

    Polymeric substrates are being identified that could permit translation of human pluripotent stem cells from laboratory-based research to industrial-scale biomedicine. Well-defined materials are required to allow cell banking and to provide the raw material for reproducible differentiation into lineages for large-scale drug-screening programs and clinical use. Yet more than 1 billion cells for each patient are needed to replace losses during heart attack, multiple sclerosis and diabetes. Producing this number of cells is challenging, and a rethink of the current predominant cell-derived substrates is needed to provide technology that can be scaled to meet the needs of millions of patients a year. In this Review, we consider the role of materials discovery, an emerging area of materials chemistry that is in large part driven by the challenges posed by biologists to materials scientists.

  9. Rapid and high-throughput construction of microbial cell-factories with regulatory noncoding RNAs.

    Science.gov (United States)

    Chaudhary, Amit Kumar; Na, Dokyun; Lee, Eun Yeol

    2015-11-01

    Due to global crises such as pollution and depletion of fossil fuels, sustainable technologies based on microbial cell-factories have been garnering great interest as an alternative to chemical factories. The development of microbial cell-factories is imperative in cutting down the overall manufacturing cost. Thus, diverse metabolic engineering strategies and engineering tools have been established to obtain a preferred genotype and phenotype displaying superior productivity. However, these tools are limited to only a handful of genes with permanent modification of a genome and significant labor costs, and this is one of the bottlenecks associated with biofactory construction. Therefore, a groundbreaking rapid and high-throughput engineering tool is needed for efficient construction of microbial cell-factories. During the last decade, copious small noncoding RNAs (ncRNAs) have been discovered in bacteria. These are involved in substantial regulatory roles like transcriptional and post-transcriptional gene regulation by modulating mRNA elongation, stability, or translational efficiency. Because of their vulnerability, ncRNAs can be used as another layer of conditional control over gene expression without modifying chromosomal sequences, and hence would be a promising high-throughput tool for metabolic engineering. Here, we review successful design principles and applications of ncRNAs for high-throughput metabolic engineering or physiological studies of diverse industrially important microorganisms.

  10. Design, Modeling, and Development of Microbial Cell Factories

    KAUST Repository

    Kodzius, Rimantas

    2014-03-26

    Using Metagenomic analysis, computational modeling, single cell and genome editing technologies, we will express desired microbial genes and their networks in suitable hosts for mass production of energy, food, and fine chemicals.

  11. Acceleration of cell factories engineering using CRISPR-based technologies

    DEFF Research Database (Denmark)

    Ronda, Carlotta

    potentially be standardized in an automatable platform and, in the future be integrated with metabolic modeling tools. In particularly it describes the technologies developed in the three widely used organisms: E. coli, S. cerevisiae and CHO mammalian cells using the recent breakthrough CRISPR/ Cas9 system....... These include CRMAGE, a MAGE improved recombineering platform using CRISPR negative selection, CrEdit, a system for multi-loci marker-free simultaneous gene and pathway integrations and CRISPy a platform to accelerate genome editing in CHO cells....

  12. Development of tools for quantitative intracellular metabolomics of Aspergillus niger chemostat cultures

    NARCIS (Netherlands)

    Lameiras, F.; Heijnen, J.J.; Van Gulik, W.M.

    2015-01-01

    In view of the high citric acid production capacity of Aspergillus niger, it should be well suited as a cell factory for the production of other relevant acids as succinic, fumaric, itaconic and malic. Quantitative metabolomics is an important omics tool in a synthetic biology approach to develop A.

  13. Novel Methods for Development of Cell Factories and Laboratory Simultations

    DEFF Research Database (Denmark)

    Bonde, Mads

    of thousands of genomic sites simultaneously. We demonstrated the feasibility of the method by inserting T7 promoters upstream of 2587 operons in E. coli, and validating the resulting cell library by deep sequencing. The method makes large-scale mutagenesis projects possible at a cost between 10x to more than...... fluorescence-activated cell sorting and deep DNA sequencing. Based on this data, we developed an algorithm (EMOPEC), which enables efficient modulation of the expression level of any chromosomally encoded gene in E. coli, by changing only a few bases in the SD sequence. We tested the algorithm by modulating 6...... ALE-acquired alleles in random combinations into the ancestral strain, and competition between these engineered strains was used to identify causative mutations by selection at 42 °C and subsequent deep sequencing of several selected clones. Interestingly, most of the identified key genes differed...

  14. Pseudomonas putida as a microbial cell factory

    DEFF Research Database (Denmark)

    Wigneswaran, Vinoth

    for sustainable production of chemicals, which can be achieved by microbial cell factories. The work presented in this PhD thesis elucidates the application of Pseudomonas putida as a microbial cell factory for production of the biosurfactant rhamnolipid. The rhamnolipid production was achieved by heterologous....... The applicability of glycerol as a substrate was also investigated. Since glycerol is a poor substrate adaptive evolution was made in order to improve the capabilities of P. putida to proliferate on glycerol. The evolved lineages all had significantly increased growth rate, enhanced cell density and reduced lag...... phase. The genomic alterations were identified by genome sequencing and revealed parallel evolution. Glycerol was also shown to be able to support biofilm growth and as a result of this it can be used as an alternative substrate for producing biochemicals in conventional and biofilm reactors. The use...

  15. Bacillus subtilis: from soil bacterium to super-secreting cell factory

    Directory of Open Access Journals (Sweden)

    van Dijl Jan Maarten

    2013-01-01

    Full Text Available Abstract The biotechnology industry has become a key element in modern societies. Within this industry, the production of recombinant enzymes and biopharmaceutical proteins is of major importance. The global markets for such recombinant proteins are growing rapidly and, accordingly, there is a continuous need for new production platforms that can deliver protein products in greater yields, with higher quality and at lower costs. This calls for the development of next-generation super-secreting cell factories. One of the microbial cell factories that can meet these challenges is the Gram-positive bacterium Bacillus subtilis, an inhabitant of the upper layers of the soil that has the capacity to secrete proteins in the gram per litre range. The engineering of B. subtilis into a next-generation super-secreting cell factory requires combined Systems and Synthetic Biology approaches. In this way, the bacterial protein secretion machinery can be optimized from the single molecule to the network level while, at the same time, taking into account the balanced use of cellular resources. Although highly ambitious, this is an achievable objective due to recent advances in functional genomics and Systems- and Synthetic Biological analyses of B. subtilis cells.

  16. Gliotoxin Isolated from Marine Fungus Aspergillus sp. Induces Apoptosis of Human Cervical Cancer and Chondrosarcoma Cells

    Directory of Open Access Journals (Sweden)

    Van-Tinh Nguyen

    2013-12-01

    Full Text Available Gliotoxin, a secondary metabolite produced by marine fungus Aspergillus sp., possesses various biological activities including anticancer activity. However, the mechanism underlying gliotoxin-induced cytotoxicity on human cervical cancer (Hela and human chondrosarcoma (SW1353 cells remains unclear. In this study, we focused on the effect of gliotoxin induction on apoptosis, the activating expressions of caspase family enzymes in the cells. Apoptotic cell levels were measured through DAPI and Annexin V/Propidium Iodide (PI double staining analysis. The apoptotic protein expression of Bcl-2 and caspase family was detected by Western blot in Hela and SW1353 cells. Our results showed that gliotoxin treatment inhibited cell proliferation and induced significant morphological changes. Gliotoxin induced apoptosis was further confirmed by DNA fragmentation, chromatin condensation and disrupted mitochondrial membrane potential. Gliotoxin-induced activation of caspase-3, caspase-8 and caspase-9, down-regulation of Bcl-2, up-regulation of Bax and cytochromec (cyt c release showed evidence for the gliotoxin activity on apoptosis. These findings suggest that gliotoxin isolated from marine fungus Aspergillus sp. induced apoptosis in Hela and SW1353 cells via the mitochondrial pathway followed by downstream events leading to apoptotic mode of cell death.

  17. CRISPR/Cas9-mediated genome engineering of CHO cell factories: application and perspectives

    DEFF Research Database (Denmark)

    Lee, Jae Seong; Grav, Lise Marie; Lewis, Nathan E.

    2015-01-01

    repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system enables rapid,easy and efficient engineering of mammalian genomes. It has a wide range of applications frommodification of individual genes to genome-wide screening or regulation of genes. Facile genomeediting using CRISPR/Cas9 empowers...... researchers in the CHO community to elucidate the mechanisticbasis behind high level production of proteins and product quality attributes of interest. Inthis review, we describe the basis of CRISPR/Cas9-mediated genome editing and its applicationfor development of next generation CHO cell factories while...... highlighting both future perspectivesand challenges. As one of the main drivers for the CHO systems biology era, genome engineeringwith CRISPR/Cas9 will pave the way for rational design of CHO cell factories....

  18. Industrial systems biology and its impact on synthetic biology of yeast cell factories

    DEFF Research Database (Denmark)

    Fletcher, Eugene; Krivoruchko, Anastasia; Nielsen, Jens

    2016-01-01

    Engineering industrial cell factories to effectively yield a desired product while dealing with industrially relevant stresses is usually the most challenging step in the development of industrial production of chemicals using microbial fermentation processes. Using synthetic biology tools......, microbial cell factories such as Saccharomyces cerevisiae can be engineered to express synthetic pathways for the production of fuels, biopharmaceuticals, fragrances, and food flavors. However, directing fluxes through these synthetic pathways towards the desired product can be demanding due to complex...... regulation or poor gene expression. Systems biology, which applies computational tools and mathematical modeling to understand complex biological networks, can be used to guide synthetic biology design. Here, we present our perspective on how systems biology can impact synthetic biology towards the goal...

  19. Systems biology of yeast: enabling technology for development of cell factories for production of advanced biofuels.

    Science.gov (United States)

    de Jong, Bouke; Siewers, Verena; Nielsen, Jens

    2012-08-01

    Transportation fuels will gradually shift from oil based fuels towards alternative fuel resources like biofuels. Current bioethanol and biodiesel can, however, not cover the increasing demand for biofuels and there is therefore a need for advanced biofuels with superior fuel properties. Novel cell factories will provide a production platform for advanced biofuels. However, deep cellular understanding is required for improvement of current biofuel cell factories. Fast screening and analysis (-omics) methods and metabolome-wide mathematical models are promising techniques. An integrated systems approach of these techniques drives diversity and quantity of several new biofuel compounds. This review will cover the recent technological developments that support improvement of the advanced biofuels 1-butanol, biodiesels and jetfuels.

  20. Production of fatty acid-derived oleochemicals and biofuels by synthetic yeast cell factories

    DEFF Research Database (Denmark)

    Zhou, Yongjin J.; Buijs, Nicolaas A; Zhu, Zhiwei

    2016-01-01

    -level production of free fatty acids (FFAs) in a yeast cell factory, and the production of alkanes and fatty alcohols from its descendants. The engineered strain produces up to 10.4 g l−1 of FFAs, which is the highest reported titre to date. Furthermore, through screening of specific pathway enzymes, endogenous...... alcohol dehydrogenases and aldehyde reductases, we reconstruct efficient pathways for conversion of fatty acids to alkanes (0.8 mg l−1) and fatty alcohols (1.5 g l−1), to our knowledge the highest titres reported in S. cerevisiae. This should facilitate the construction of yeast cell factories...... for production of fatty acids derived products and even aldehyde-derived chemicals of high value....

  1. Disseminated aspergillosis caused by Aspergillus ustus in a patient following allogeneic peripheral stem cell transplantation.

    Science.gov (United States)

    Iwen, P C; Rupp, M E; Bishop, M R; Rinaldi, M G; Sutton, D A; Tarantolo, S; Hinrichs, S H

    1998-12-01

    The first case of disseminated aspergillosis caused by Aspergillus ustus in an allogeneic peripheral stem cell transplant patient is described. The patient, a 46-year-old female with a history of myelodysplastic syndrome, underwent high-dose chemotherapy and total body irradiation prior to transplantation. She was released from the hospital 49 days posttransplant (p.t.) in a stable condition with an absolute neutrophil count (ANC) of 2,700 cells per microl. Multiple antimicrobial agents, including itraconazole (ITR), were prescribed during hospitalization and at the time of discharge. Three days after discharge, the patient was readmitted with hemorrhagic cystitis, persistent thrombocytopenia, and bilateral pulmonary consolidation, although no fever was present. The ANC at the time of readmission was 3,500. Upon detection of a pulmonary nodule (day 67 p.t.), a bronchoalveolar lavage was performed; the lavage fluid was positive for both cytomegalovirus and parainfluenza virus and negative for fungus. The patient was placed on ganciclovir. A biopsy specimen from a leg lesion also noted on day 67 p.t. revealed septate hyphae consistent with Aspergillus species, and a culture subsequently yielded Aspergillus ustus. Confirmation detection of A. ustus was made by demonstration of characteristic reproductive structures with the presence of Hülle cells. On day 67 p.t., ITR was discontinued and liposomal amphotericin B (AMB) was initiated. The patient's condition worsened, and she died 79 days p.t. At the time of autopsy, septate hyphae were present in heart, thyroid, and lung tissues, with lung tissue culture positive for A. ustus. In vitro susceptibility testing indicated probable resistance to AMB but not to ITR. This case supports the need for the development of rapid methods to determine antifungal susceptibility.

  2. Expression and export: recombinant protein production systems for Aspergillus.

    Science.gov (United States)

    Fleissner, André; Dersch, Petra

    2010-07-01

    Several Aspergillus species, in particular Aspergillus niger and Aspergillus oryzae, are widely used as protein production hosts in various biotechnological applications. In order to improve the expression and secretion of recombinant proteins in these filamentous fungi, several novel genetic engineering strategies have been developed in recent years. This review describes state-of-the-art genetic manipulation technologies used for strain improvement, as well as recent advances in designing the most appropriate engineering strategy for a particular protein production process. Furthermore, current developments in identifying bottlenecks in the protein production and secretion pathways are described and novel approaches to overcome these limitations are introduced. An appropriate combination of expression vectors and optimized host strains will provide cell factories customized for each production process and expand the great potential of Aspergilli as biotechnology workhorses to more complex multi-step industrial applications.

  3. Aspergillus enzymes involved in degradation of plant cell wall polysaccharides

    NARCIS (Netherlands)

    Vries, de R.P.; Visser, J.

    2001-01-01

    Degradation of plant cell wall polysaccharides is of major importance in the food and feed, beverage, textile, and paper and pulp industries, as well as in several other industrial production processes. Enzymatic degradation of these polymers has received attention for many years and is becoming a m

  4. Live-cell imaging of Aspergillus nidulans autophagy: RAB1 dependence, Golgi independence and ER involvement

    OpenAIRE

    Pinar, Mario; Pantazopoulou, Areti; Peñalva, Miguel A.

    2013-01-01

    We exploited the amenability of the fungus Aspergillus nidulans to genetics and live-cell microscopy to investigate autophagy. Upon nitrogen starvation, GFP-Atg8-containing pre-autophagosomal puncta give rise to cup-shaped phagophores and circular (0.9-μm diameter) autophagosomes that disappear in the vicinity of the vacuoles after their shape becomes irregular and their GFP-Atg8 fluorescence decays. This ‘autophagosome cycle’ gives rise to characteristic cone-shaped traces in kymographs. Aut...

  5. Aspergillus Cell Wall Melanin Blocks LC3-Associated Phagocytosis to Promote Pathogenicity.

    Science.gov (United States)

    Akoumianaki, Tonia; Kyrmizi, Irene; Valsecchi, Isabel; Gresnigt, Mark S; Samonis, George; Drakos, Elias; Boumpas, Dimitrios; Muszkieta, Laetitia; Prevost, Marie-Christine; Kontoyiannis, Dimitrios P; Chavakis, Triantafyllos; Netea, Mihai G; van de Veerdonk, Frank L; Brakhage, Axel A; El-Benna, Jamel; Beauvais, Anne; Latge, Jean-Paul; Chamilos, Georgios

    2016-01-13

    Concealing pathogen-associated molecular patterns (PAMPs) is a principal strategy used by fungi to avoid immune recognition. Surface exposure of PAMPs during germination can leave the pathogen vulnerable. Accordingly, β-glucan surface exposure during Aspergillus fumigatus germination activates an Atg5-dependent autophagy pathway termed LC3-associated phagocytosis (LAP), which promotes fungal killing. We found that LAP activation also requires the genetic, biochemical or biological (germination) removal of A. fumigatus cell wall melanin. The attenuated virulence of melanin-deficient A. fumigatus is restored in Atg5-deficient macrophages and in mice upon conditional inactivation of Atg5 in hematopoietic cells. Mechanistically, Aspergillus melanin inhibits NADPH oxidase-dependent activation of LAP by excluding the p22phox subunit from the phagosome. Thus, two events that occur concomitantly during germination of airborne fungi, surface exposure of PAMPs and melanin removal, are necessary for LAP activation and fungal killing. LAP blockade is a general property of melanin pigments, a finding with broad physiological implications.

  6. CRISPR/Cas9-mediated genome engineering of CHO cell factories: Application and perspectives.

    Science.gov (United States)

    Lee, Jae Seong; Grav, Lise Marie; Lewis, Nathan E; Faustrup Kildegaard, Helene

    2015-07-01

    Chinese hamster ovary (CHO) cells are the most widely used production host for therapeutic proteins. With the recent emergence of CHO genome sequences, CHO cell line engineering has taken on a new aspect through targeted genome editing. The bacterial clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system enables rapid, easy and efficient engineering of mammalian genomes. It has a wide range of applications from modification of individual genes to genome-wide screening or regulation of genes. Facile genome editing using CRISPR/Cas9 empowers researchers in the CHO community to elucidate the mechanistic basis behind high level production of proteins and product quality attributes of interest. In this review, we describe the basis of CRISPR/Cas9-mediated genome editing and its application for development of next generation CHO cell factories while highlighting both future perspectives and challenges. As one of the main drivers for the CHO systems biology era, genome engineering with CRISPR/Cas9 will pave the way for rational design of CHO cell factories.

  7. Rapid prototyping of microbial cell factories via genome-scale engineering.

    Science.gov (United States)

    Si, Tong; Xiao, Han; Zhao, Huimin

    2015-11-15

    Advances in reading, writing and editing genetic materials have greatly expanded our ability to reprogram biological systems at the resolution of a single nucleotide and on the scale of a whole genome. Such capacity has greatly accelerated the cycles of design, build and test to engineer microbes for efficient synthesis of fuels, chemicals and drugs. In this review, we summarize the emerging technologies that have been applied, or are potentially useful for genome-scale engineering in microbial systems. We will focus on the development of high-throughput methodologies, which may accelerate the prototyping of microbial cell factories.

  8. Production of plant cell wall degrading enzymes by monoculture and co-culture of Aspergillus niger and Aspergillus terreus under SSF of banana peels.

    Science.gov (United States)

    Rehman, Shazia; Aslam, Hina; Ahmad, Aqeel; Khan, Shakeel Ahmed; Sohail, Muhammad

    2014-01-01

    Filamentous fungi are considered to be the most important group of microorganisms for the production of plant cell wall degrading enzymes (CWDE), in solid state fermentations. In this study, two fungal strains Aspergillus niger MS23 and Aspergillus terreus MS105 were screened for plant CWDE such as amylase, pectinase, xylanase and cellulases (β-glucosidase, endoglucanase and filterpaperase) using a novel substrate, Banana Peels (BP) for SSF process. This is the first study, to the best of our knowledge, to use BP as SSF substrate for plant CWDE production by co-culture of fungal strains. The titers of pectinase were significantly improved in co-culture compared to mono-culture. Furthermore, the enzyme preparations obtained from monoculture and co-culture were used to study the hydrolysis of BP along with some crude and purified substrates. It was observed that the enzymatic hydrolysis of different crude and purified substrates accomplished after 26 h of incubation, where pectin was maximally hydrolyzed by the enzyme preparations of mono and co-culture. Along with purified substrates, crude materials were also proved to be efficiently degraded by the cocktail of the CWDE. These results demonstrated that banana peels may be a potential substrate in solid-state fermentation for the production of plant cell wall degrading enzymes to be used for improving various biotechnological and industrial processes.

  9. Production of plant cell wall degrading enzymes by monoculture and co-culture of Aspergillus niger and Aspergillus terreus under SSF of banana peels

    Directory of Open Access Journals (Sweden)

    Shazia Rehman

    2014-12-01

    Full Text Available Filamentous fungi are considered to be the most important group of microorganisms for the production of plant cell wall degrading enzymes (CWDE, in solid state fermentations. In this study, two fungal strains Aspergillus niger MS23 and Aspergillus terreus MS105 were screened for plant CWDE such as amylase, pectinase, xylanase and cellulases (β-glucosidase, endoglucanase and filterpaperase using a novel substrate, Banana Peels (BP for SSF process. This is the first study, to the best of our knowledge, to use BP as SSF substrate for plant CWDE production by co-culture of fungal strains. The titers of pectinase were significantly improved in co-culture compared to mono-culture. Furthermore, the enzyme preparations obtained from monoculture and co-culture were used to study the hydrolysis of BP along with some crude and purified substrates. It was observed that the enzymatic hydrolysis of different crude and purified substrates accomplished after 26 h of incubation, where pectin was maximally hydrolyzed by the enzyme preparations of mono and co-culture. Along with purified substrates, crude materials were also proved to be efficiently degraded by the cocktail of the CWDE. These results demonstrated that banana peels may be a potential substrate in solid-state fermentation for the production of plant cell wall degrading enzymes to be used for improving various biotechnological and industrial processes.

  10. Isolate-dependent growth, virulence, and cell wall composition in the human pathogen Aspergillus fumigatus.

    Directory of Open Access Journals (Sweden)

    Nansalmaa Amarsaikhan

    Full Text Available The ubiquitous fungal pathogen Aspergillus fumigatus is a mediator of allergic sensitization and invasive disease in susceptible individuals. The significant genetic and phenotypic variability between and among clinical and environmental isolates are important considerations in host-pathogen studies of A. fumigatus-mediated disease. We observed decreased radial growth, rate of germination, and ability to establish colony growth in a single environmental isolate of A. fumigatus, Af5517, when compared to other clinical and environmental isolates. Af5517 also exhibited increased hyphal diameter and cell wall β-glucan and chitin content, with chitin most significantly increased. Morbidity, mortality, lung fungal burden, and tissue pathology were decreased in neutropenic Af5517-infected mice when compared to the clinical isolate Af293. Our results support previous findings that suggest a correlation between in vitro growth rates and in vivo virulence, and we propose that changes in cell wall composition may contribute to this phenotype.

  11. Spinal osteomyelitis due to Aspergillus flavus in a child: a rare complication after haematopoietic stem cell transplantation

    Energy Technology Data Exchange (ETDEWEB)

    Beluffi, Giampiero [Fondazione IRCCS Policlinico ' S.Matteo' , Section of Paediatric Radiology, Department of Radiodiagnosis, Pavia PV (Italy); Bernardo, Maria E.; Locatelli, Franco [University of Pavia, Fondazione IRCCS Policlinico ' S.Matteo' , Department of Paediatric Haematology/Oncology, Pavia (Italy); Meloni, Giulia [University of Pavia, Fondazione IRCCS Policlinico ' S.Matteo' , Institute of Radiology, Pavia (Italy); Spinazzola, Angelo [Fondazione IRCCS Policlinico ' S.Matteo' , Section of Paediatric Radiology, Department of Radiodiagnosis, Pavia PV (Italy); Ospedale Maggiore, Crema CR (Italy)

    2008-06-15

    We report the case of a child affected by acute myeloid leukaemia who was treated with allogeneic haematopoietic stem cell transplantation and developed cervicothoracic spinal osteomyelitis due to Aspergillus flavus. The diagnosis was difficult on a clinical basis, but made possible by conventional radiography and MRI. (orig.)

  12. ESCHERICHIA COLI REDOX MUTANTS AS MICROBIAL CELL FACTORIES FOR THE SYNTHESIS OF REDUCED BIOCHEMICALS

    Directory of Open Access Journals (Sweden)

    Jimena A. Ruiz

    2012-10-01

    Full Text Available Bioprocesses conducted under conditions with restricted O2 supply are increasingly exploited for the synthesis of reduced biochemicals using different biocatalysts. The model facultative aerobe Escherichia coli, the microbial cell factory par excellence, has elaborate sensing and signal transduction mechanisms that respond to the availability of electron acceptors and alternative carbon sources in the surrounding environment. In particular, the ArcBA and CreBC two-component signal transduction systems are largely responsible for the metabolic regulation of redox control in response to O2 availability and carbon source utilization, respectively. Significant advances in the understanding of the biochemical, genetic, and physiological duties of these regulatory systems have been achieved in recent years. This situation allowed to rationally-design novel engineering approaches that ensure optimal carbon and energy flows within central metabolism, as well as to manipulate redox homeostasis, in order to optimize the production of industrially-relevant metabolites. In particular, metabolic flux analysis provided new clues to understand the metabolic regulation mediated by the ArcBA and CreBC systems. Genetic manipulation of these regulators proved useful for designing microbial cells factories tailored for the synthesis of reduced biochemicals with added value, such as poly(3-hydroxybutyrate, under conditions with restricted O2 supply. This network-wide strategy is in contrast with traditional metabolic engineering approaches, that entail direct modification of the pathway(s at stake, and opens new avenues for the targeted modulation of central catabolic pathways at the transcriptional level.

  13. Human Invariant Natural Killer T cells possess immune-modulating functions during Aspergillus infection.

    Science.gov (United States)

    Beitzen-Heineke, Antonia; Bouzani, Maria; Schmitt, Anna-Lena; Kurzai, Oliver; Hünniger, Kerstin; Einsele, Hermann; Loeffler, Juergen

    2016-02-01

    Aspergillus fumigatus is the most common cause for invasive fungal infections, a disease associated with high mortality in immune-compromised patients. CD1d-restricted invariant natural killer T (iNKT) cells compose a small subset of T cells known to impact the immune response toward various infectious pathogens. To investigate the role of human iNKT cells during A. fumigatus infection, we studied their activation as determined by CD69 expression and cytokine production in response to distinct fungal morphotypes in the presence of different CD1d(+) antigen presenting cells using flow cytometry and multiplex enzyme-linked immunosorbent assay (ELISA). Among CD1d(+) subpopulations, CD1d(+)CD1c(+) mDCs showed the highest potential to activate iNKT cells on a per cell basis. The presence of A. fumigatus decreased this effect of CD1d(+)CD1c(+) mDCs on iNKT cells and led to reduced secretion of TNF-α, G-CSF and RANTES. Production of other Th1 and Th2 cytokines was not affected by the fungus, suggesting an immune-modulating function for human iNKT cells during A. fumigatus infection.

  14. Targeting population heterogeneity in Saccharomyces cerevisiae batch fermentation for optimal cell factories

    DEFF Research Database (Denmark)

    Heins, Anna-Lena; Lencastre Fernandes, Rita; Lundin, L.

    constructed which enabled us to perform single cell level analysis, and thereby created the possibility to map population heterogeneity. A factorial design with pH, glucose concentration and oxygen level was performed in batch cultivations using the growth reporter strains to evaluate the effect of those...... environmental factors on heterogeneity level and amount of living cells. A highly dynamic behavior with regard to subpopulation distribution during the different growth stages was seen for the batch cultivations. Moreover, it could be demonstrated that the glucose concentration had a clear influence......To achieve an efficient production process, it is essential to optimize both the strain and the cultivation conditions. Traditionally, a microbial population has been considered homogeneous in optimization studies of fermentation processes. However, research has shown that a typical microbial...

  15. Aspergillus nidulans cell wall composition and function change in response to hosting several Aspergillus fumigatus UDP-galactopyranose mutase activity mutants.

    Directory of Open Access Journals (Sweden)

    Md Kausar Alam

    Full Text Available Deletion or repression of Aspergillus nidulans ugmA (AnugmA, involved in galactofuranose biosynthesis, impairs growth and increases sensitivity to Caspofungin, a β-1,3-glucan synthesis antagonist. The A. fumigatus UgmA (AfUgmA crystal structure has been determined. From that study, AfUgmA mutants with altered enzyme activity were transformed into AnugmA▵ to assess their effect on growth and wall composition in A. nidulans. The complemented (AnugmA::wild type AfugmA strain had wild type phenotype, indicating these genes had functional homology. Consistent with in vitro studies, AfUgmA residues R182 and R327 were important for its function in vivo, with even conservative amino (RK substitutions producing AnugmA? phenotype strains. Similarly, the conserved AfUgmA loop III histidine (H63 was important for Galf generation: the H63N strain had a partially rescued phenotype compared to AnugmA▵. Collectively, A. nidulans strains that hosted mutated AfUgmA constructs with low enzyme activity showed increased hyphal surface adhesion as assessed by binding fluorescent latex beads. Consistent with previous qPCR results, immunofluorescence and ELISA indicated that AnugmA▵ and AfugmA-mutated A. nidulans strains had increased α-glucan and decreased β-glucan in their cell walls compared to wild type and AfugmA-complemented strains. Like the AnugmA▵ strain, A. nidulans strains containing mutated AfugmA showed increased sensitivity to antifungal drugs, particularly Caspofungin. Reduced β-glucan content was correlated with increased Caspofungin sensitivity. Aspergillus nidulans wall Galf, α-glucan, and β-glucan content was correlated in A. nidulans hyphal walls, suggesting dynamic coordination between cell wall synthesis and cell wall integrity.

  16. Aspergillus nidulans cell wall composition and function change in response to hosting several Aspergillus fumigatus UDP-galactopyranose mutase activity mutants.

    Science.gov (United States)

    Alam, Md Kausar; van Straaten, Karin E; Sanders, David A R; Kaminskyj, Susan G W

    2014-01-01

    Deletion or repression of Aspergillus nidulans ugmA (AnugmA), involved in galactofuranose biosynthesis, impairs growth and increases sensitivity to Caspofungin, a β-1,3-glucan synthesis antagonist. The A. fumigatus UgmA (AfUgmA) crystal structure has been determined. From that study, AfUgmA mutants with altered enzyme activity were transformed into AnugmA▵ to assess their effect on growth and wall composition in A. nidulans. The complemented (AnugmA::wild type AfugmA) strain had wild type phenotype, indicating these genes had functional homology. Consistent with in vitro studies, AfUgmA residues R182 and R327 were important for its function in vivo, with even conservative amino (RK) substitutions producing AnugmA? phenotype strains. Similarly, the conserved AfUgmA loop III histidine (H63) was important for Galf generation: the H63N strain had a partially rescued phenotype compared to AnugmA▵. Collectively, A. nidulans strains that hosted mutated AfUgmA constructs with low enzyme activity showed increased hyphal surface adhesion as assessed by binding fluorescent latex beads. Consistent with previous qPCR results, immunofluorescence and ELISA indicated that AnugmA▵ and AfugmA-mutated A. nidulans strains had increased α-glucan and decreased β-glucan in their cell walls compared to wild type and AfugmA-complemented strains. Like the AnugmA▵ strain, A. nidulans strains containing mutated AfugmA showed increased sensitivity to antifungal drugs, particularly Caspofungin. Reduced β-glucan content was correlated with increased Caspofungin sensitivity. Aspergillus nidulans wall Galf, α-glucan, and β-glucan content was correlated in A. nidulans hyphal walls, suggesting dynamic coordination between cell wall synthesis and cell wall integrity.

  17. Inducible expression of beta defensins by human respiratory epithelial cells exposed to Aspergillus fumigatus organisms

    Directory of Open Access Journals (Sweden)

    Tichanné-Seltzer Virginie

    2009-02-01

    Full Text Available Abstract Background Aspergillus fumigatus, a saprophytic mould, is responsible for life-threatening, invasive pulmonary diseases in immunocompromised hosts. The role of the airway epithelium involves a complex interaction with the inhaled pathogen. Antimicrobial peptides with direct antifungal and chemotactic activities may boost antifungal immune response. Results The inducible expression of defensins by human bronchial epithelial 16HBE cells and A549 pneumocyte cells exposed to A. fumigatus was investigated. Using RT-PCR and real time PCR, we showed an activation of hBD2 and hBD9 defensin genes: the expression was higher in cells exposed to swollen conidia (SC, compared to resting conidia (RC or hyphal fragments (HF. The kinetics of defensin expression was different for each one, evoking a putative distinct function for each investigated defensin. The decrease of defensin expression in the presence of heat-inactivated serum indicated a possible link between defensins and the proteins of the host complement system. The presence of defensin peptide hBD2 was revealed using immunofluorescence that showed a punctual cytoplasmic and perinuclear staining. Quantification of the cells stained with anti hBD2 antibody demonstrated that SC induced a greater number of cells that synthesized hBD2, compared to RC or HF. Labelling of the cells with anti-hBD-2 antibody showed a positive immunofluorescence signal around RC or SC in contrast to HF. This suggests co-localisation of hBD2 and digested conidia. The HBD2 level was highest in the supernatants of cells exposed to SC, as was determined by sandwich ELISA. Experiments using neutralising anti-interleukine-1β antibody reflect the autocrine mechanism of defensin expression induced by SC. Investigation of defensin expression at transcriptional and post-transcriptional levels demonstrated the requirement of transcription as well as new protein synthesis during A. fumigatus defensin induction. Finally, induced

  18. E-cadherin mediates adhesion and endocytosis of Aspergillus fumigatus blastospores in human epithelial cells

    Institute of Scientific and Technical Information of China (English)

    XU Xiao-yong; SHI Yi; ZHANG Peng-peng; ZHANG Feng; SHEN Yu-ying; SU Xin; ZHAO Bei-lei

    2012-01-01

    Background Aspergillus fumigatus (A.fumigatus) is a ubiquitous saprophytic fungus responsible for the majority of invasive mold infections in patients undergoing chemotherapy,organ transplantation or with persistent neutropenia.This study aimed to determine the role of E-cadherin for adhesion and endocytosis of A.fumigatus blastospores in the human epithelial cell line A549.Methods A.fumigatus blastospores were incubated with the total protein of A549 to investigate the binding of E-cadherin and blastospores followed by an affinity purification procedure.After establishing the adhesion model,the adhesion and endocytosis of A.fumigatus blastospores by A549 cells were evaluated by down-regulating E-cadherin of A549 cells using blocking antibody or small interfering RNA (siRNA).Results E-cadherin was adhered to the surface of A.fumigatus blastospore.Adhesion and endocytosis of the blastospores were reduced by blocking or down-regulating E-cadherin in A549 cells.Conclusions E-cadherin is a receptor for adhesion and endocytosis of A.fumigatus blastospores in epithelial cells.This may open a new approach to treat this fungal infection.

  19. Th and Treg response induced by Aspergillus fumigatus pulsed dendritic cells in vitro

    Institute of Scientific and Technical Information of China (English)

    Wang Runchao; Wan Zhe; Li Ruoyu

    2014-01-01

    Background Dendritic cells (DCs) can recognize the pathogen-associated molecular patterns (PAMP) of Aspergillus fumigatus (A.fumigatus),activating the immune response.During A.fumigatus infection,a Th and Treg response induced in the fungi-pulsed DCs is not yet well understood.Methods In this study,bone marrow-derived dendritic cells (BMDCs) were separated and proliferated from C57BL/6 mice.A.fumigatus pulsed DCs were generated and cultured with CD4+ T cells derived from the spleen of C57BL/6 mice in vitro.CD4+ T cells differentiation after co-culture were analyzed by flow cytometry,ELISA,and real-time PCR analysis.Results The A.fumigatus pulsed DCs exhibited increased Th1 and Treg frequency,Th1-related cytokines (IFN-γ and IL-12),Treg-related cytokines (TGF-β) and T-bet,and Foxp3 mRNA levels compared with the control group.There was no significant difference between A.fumigatus pulsed DCs group and the control group about Th17 and Th2 frequency.Conclusions The inactivated conidia of A.fumigatus were able to activate BMDCs and made them capable of triggering T cell responses in vitro.A.fumigatus loaded DCs was a weak inducer of Th17 and Th2,but induced a strong Th1 and Treg response.

  20. Development of a screening approach for exploring cell factory potential through metabolic flux analysis and physiology

    DEFF Research Database (Denmark)

    Knudsen, Peter Boldsen; Nielsen, Kristian Fog; Thykær, Jette

    2012-01-01

    The recent developments within the field of metabolic engineering have significantly increased the speed by which fungal recombinant strains are being constructed, pushing focus towards physiological characterisation and analysis. This raises demand for a tool for diligent analysis of the recombi...... and work-load connected with screening and selection of potential cell factories with attractive properties, compared with more “traditional” methodologies where metabolic flux analysis is applied at a much later state in the characterisation process.......The recent developments within the field of metabolic engineering have significantly increased the speed by which fungal recombinant strains are being constructed, pushing focus towards physiological characterisation and analysis. This raises demand for a tool for diligent analysis...... on a Hamilton robotic system. This method aimed at characterising physiology at two levels: (1) An approach focusing on the traditional growth related parameters, i.e. growth rate, yield coefficients and extracellular metabolites. (2) 13C-labelling experiments, where metabolic fluxes are quantified...

  1. In Silico Constraint-Based Strain Optimization Methods: the Quest for Optimal Cell Factories.

    Science.gov (United States)

    Maia, Paulo; Rocha, Miguel; Rocha, Isabel

    2016-03-01

    Shifting from chemical to biotechnological processes is one of the cornerstones of 21st century industry. The production of a great range of chemicals via biotechnological means is a key challenge on the way toward a bio-based economy. However, this shift is occurring at a pace slower than initially expected. The development of efficient cell factories that allow for competitive production yields is of paramount importance for this leap to happen. Constraint-based models of metabolism, together with in silico strain design algorithms, promise to reveal insights into the best genetic design strategies, a step further toward achieving that goal. In this work, a thorough analysis of the main in silico constraint-based strain design strategies and algorithms is presented, their application in real-world case studies is analyzed, and a path for the future is discussed.

  2. Thermodynamics-based design of microbial cell factories for anaerobic product formation.

    Science.gov (United States)

    Cueto-Rojas, Hugo F; van Maris, A J A; Wahl, S Aljoscha; Heijnen, J J

    2015-09-01

    The field of metabolic engineering has delivered new microbial cell factories and processes for the production of different compounds including biofuels, (di)carboxylic acids, alcohols, and amino acids. Most of these processes are aerobic, with few exceptions (e.g., alcoholic fermentation), and attention is focused on assembling a high-flux product pathway with a production limit usually set by the oxygen transfer rate. By contrast, anaerobic product synthesis offers significant benefits compared to aerobic systems: higher yields, less heat generation, reduced biomass production, and lower mechanical energy input, which can significantly reduce production costs. Using simple thermodynamic calculations, we demonstrate that many products can theoretically be produced under anaerobic conditions using several conventional and non-conventional substrates.

  3. A novel cell factory for efficient production of ethanol from dairy waste

    DEFF Research Database (Denmark)

    Liu, Jianming; Dantoft, Shruti Harnal; Würtz, Anders

    2016-01-01

    if cost-effective processes are to be established. Various industries, especially within the food sector, often have waste streams rich in carbohydrates and/or other nutrients, and these could serve as alternative feedstocks for such bio-processes. The dairy industry is a good example, where large amounts...... and focus has mostly been on developing bio-processes involving non-food plant biomass feedstocks. The high cost of the enzymes needed to degrade such feedstocks into its constituent sugars as well as problems due to various inhibitors generated in pretreatment are two challenges that have to be addressed...... hydrolysate (CSLH) as the nitrogen source. The outcome was efficient ethanol production with a titer of 41 g/L and a yield of 70 % of the theoretical maximum using a fed-batch strategy. The combination of a low-cost medium from industrial waste streams and an efficient cell factory should make the developed...

  4. A novel metabolite from aspergillus ochraceus JGI 25 showing cytotoxicity to hela cells

    Directory of Open Access Journals (Sweden)

    Varalakshmi K Nadumane

    2013-01-01

    Full Text Available This study aims at the isolation of filamentous fungi, extraction of metabolites, and evaluation of the cytotoxic properties on HeLa cells and normal human lymphocytes. We isolated fungi from the soil by serial dilution method. One of the isolates was chosen and identified as Aspergillus ochraceus Wilhelm (Trichocomaceae by standard techniques. The metabolites were extracted using methanol. Different concentrations of the extract were evaluated for their potential anticancer activity on HeLa cells by 3-(4,5-dimethylthiazol-2yl-2,5-diphenyl tetrazolium bromide assay and the safety of the extract was checked on normal human lymphocytes. The extract was purified by chromatographic techniques like thin-layer chromatography and high-performance liquid chromatography, and subjected to mass spectrometric analysis. The extract showed significant cytotoxic potential on HeLa cells at low concentrations with a half maximal inhibitory concentration value of <50 ΅g/ml. The extract gave 10 fractions by thin layer chromatography, and fraction B had higher toxicity than the rest. This fraction gave a single peak by high-performance liquid chromatography and had a mass-to-charge ratio of 905.65, which did not match any of the earlier known fungal metabolites or metabolites from other strains of A. ochraceus. The metabolite from A. ochraceus is alkaloid in nature, cytotoxic to HeLa cells, and appears to be a novel with anticancer potentials, which could be explored further for characterization of the active component.

  5. Nanoscale biophysical properties of the cell surface galactosaminogalactan from the fungal pathogen Aspergillus fumigatus.

    Science.gov (United States)

    Beaussart, Audrey; El-Kirat-Chatel, Sofiane; Fontaine, Thierry; Latgé, Jean-Paul; Dufrêne, Yves F

    2015-09-28

    Many fungal pathogens produce cell surface polysaccharides that play essential roles in host-pathogen interactions. In Aspergillus fumigatus, the newly discovered polysaccharide galactosaminogalactan (GAG) mediates adherence to a variety of substrates through molecular mechanisms that are poorly understood. Here we use atomic force microscopy to unravel the localization and adhesion of GAG on living fungal cells. Using single-molecule imaging with tips bearing anti-GAG antibodies, we found that GAG is massively exposed on wild-type (WT) germ tubes, consistent with the notion that this glycopolymer is secreted by the mycelium of A. fumigatus, while it is lacking on WT resting conidia and on germ tubes from a mutant (Δuge3) deficient in GAG. Imaging germ tubes with tips bearing anti-β-glucan antibodies shows that exposure of β-glucan is strongly increased in the Δuge3 mutant, indicating that this polysaccharide is masked by GAG during hyphal growth. Single-cell force measurements show that expression of GAG on germ tubes promotes specific adhesion to pneumocytes and non-specific adhesion to hydrophobic substrates. These results provide a molecular foundation for the multifunctional adhesion properties of GAG, thus suggesting it could be used as a potential target in anti-adhesion therapy and immunotherapy. Our methodology represents a powerful approach for characterizing the nanoscale organization and adhesion of cell wall polysaccharides during fungal morphogenesis, thereby contributing to increase our understanding of their role in biofilm formation and immune responses.

  6. Development of Synechocystis sp. PCC 6803 as a Phototrophic Cell Factory

    Directory of Open Access Journals (Sweden)

    Fuzhong Zhang

    2013-08-01

    Full Text Available Cyanobacteria (blue-green algae play profound roles in ecology and biogeochemistry. One model cyanobacterial species is the unicellular cyanobacterium Synechocystis sp. PCC 6803. This species is highly amenable to genetic modification. Its genome has been sequenced and many systems biology and molecular biology tools are available to study this bacterium. Recently, researchers have put significant efforts into understanding and engineering this bacterium to produce chemicals and biofuels from sunlight and CO2. To demonstrate our perspective on the application of this cyanobacterium as a photosynthesis-based chassis, we summarize the recent research on Synechocystis 6803 by focusing on five topics: rate-limiting factors for cell cultivation; molecular tools for genetic modifications; high-throughput system biology for genome wide analysis; metabolic modeling for physiological prediction and rational metabolic engineering; and applications in producing diverse chemicals. We also discuss the particular challenges for systems analysis and engineering applications of this microorganism, including precise characterization of versatile cell metabolism, improvement of product rates and titers, bioprocess scale-up, and product recovery. Although much progress has been achieved in the development of Synechocystis 6803 as a phototrophic cell factory, the biotechnology for “Compounds from Synechocystis” is still significantly lagging behind those for heterotrophic microbes (e.g., Escherichia coli.

  7. Design, Engineering, and Construction of Photosynthetic Microbial Cell Factories for Renewable Solar Fuel Production

    Energy Technology Data Exchange (ETDEWEB)

    Lindblad, Peter; Lindberg, Pia; Stensjoe, Karin (Photochemistry and Molecular Science, Dept. of Chemistry-Aangstroem Laboratory, Uppsala Univ., Uppsala (Sweden)), E-mail: Peter.Lindblad@kemi.uu.se; Oliveira, Paulo (Instituto de Biologia Molecular e Celular, Porto (Portugal)); Heidorn, Thorsten (Bioforsk-Norwegian Inst. for Agricultural and Environmental Research, Aas Oslo, (Norway))

    2012-03-15

    There is an urgent need to develop sustainable solutions to convert solar energy into energy carriers used in the society. In addition to solar cells generating electricity, there are several options to generate solar fuels. This paper outlines and discusses the design and engineering of photosynthetic microbial systems for the generation of renewable solar fuels, with a focus on cyanobacteria. Cyanobacteria are prokaryotic microorganisms with the same type of photosynthesis as higher plants. Native and engineered cyanobacteria have been used by us and others as model systems to examine, demonstrate, and develop photobiological H{sub 2} production. More recently, the production of carbon-containing solar fuels like ethanol, butanol, and isoprene have been demonstrated. We are using a synthetic biology approach to develop efficient photosynthetic microbial cell factories for direct generation of biofuels from solar energy. Present progress and advances in the design, engineering, and construction of such cyanobacterial cells for the generation of a portfolio of solar fuels, e.g., hydrogen, alcohols, and isoprene, are presented and discussed. Possibilities and challenges when introducing and using synthetic biology are highlighted

  8. Mitotic destruction of the cell cycle regulated NIMA protein kinase of Aspergillus nidulans is required for mitotic exit.

    OpenAIRE

    Pu, R T; Osmani, S A

    1995-01-01

    NIMA is a cell cycle regulated protein kinase required, in addition to p34cdc2/cyclin B, for initiation of mitosis in Aspergillus nidulans. Like cyclin B, NIMA accumulates when cells are arrested in G2 and is degraded as cells traverse mitosis. However, it is stable in cells arrested in mitosis. NIMA, and related kinases, have an N-terminal kinase domain and a C-terminal extension. Deletion of the C-terminus does not completely inactivate NIMA kinase activity but does prevent functional compl...

  9. Enantioselective hydrolysis of epichlorohydrin using whole Aspergillus niger ZJB-09173 cells in organic solvents

    Indian Academy of Sciences (India)

    Huo-Xi Jin; Zhong-Ce Hu; Yu-Guo Zheng

    2012-09-01

    The enantioselective hydrolysis of racemic epichlorohydrin for the production of enantiopure ()-epichloro-hydrin using whole cells of Aspergillus niger ZJB-09173 in organic solvents was investigated. Cyclohexane was used as the reaction medium based on the excellent enantioselectivity of epoxide hydrolase from A. niger ZJB-09173 in cyclohexane. However, cyclohexane had a negative effect on the stability of epoxide hydrolase from A. niger ZJB-09173. In the cyclohexane medium, substrate inhibition, rather than product inhibition of catalysis, was observed in the hydrolysis of racemic epichlorohydrin using A. niger ZJB-09173. The racemic epichlorohydrin concentration was markedly increased by continuous feeding of substrate without significant decline of the yield. Ultimately, 18.5% of ()-epichlorohydrin with 98% enantiomeric excess from 153.6 mM of racemic epichlorohydrin was obtained by the dry cells of A. niger ZJB-09173, which was the highest substrate concentration in the production of enantiopure ()-epichlorohydrin by epoxide hydrolases using an organic solvent medium among the known reports.

  10. Surfactant Protein-A inhibits Aspergillus fumigatus-induced allergic T-cell responses

    Directory of Open Access Journals (Sweden)

    Russo Scott J

    2005-08-01

    Full Text Available Abstract Background The pulmonary surfactant protein (SP-A has potent immunomodulatory activities but its role and regulation during allergic airway inflammation is unknown. Methods We studied changes in SP-A expression in the bronchoalveolar lavage (BAL using a murine model of single Aspergillus fumigatus (Af challenge of sensitized animals. Results SP-A protein levels in the BAL fluid showed a rapid, transient decline that reached the lowest values (25% of controls 12 h after intranasal Af provocation of sensitized mice. Decrease of SP-A was associated with influx of inflammatory cells and increase of IL-4 and IL-5 mRNA and protein levels. Since levels of SP-A showed a significant negative correlation with these BAL cytokines (but not with IFN-γ, we hypothesized that SP-A exerts an inhibitory effect on Th2-type immune responses. To study this hypothesis, we used an in vitro Af-rechallenge model. Af-induced lymphocyte proliferation of cells isolated from sensitized mice was inhibited in a dose-dependent manner by addition of purified human SP-A (0.1–10 μg/ml. Flow cytometric studies on Af-stimulated lymphocytes indicated that the numbers of CD4+ (but not CD8+ T cells were significantly increased in the parental population and decreased in the third and fourth generation in the presence of SP-A. Further, addition of SP-A to the tissue culture inhibited Af-induced IL-4 and IL-5 production suggesting that SP-A directly suppressed allergen-stimulated CD4+ T cell function. Conclusion We speculate that a transient lack of this lung collectin following allergen exposure of the airways may significantly contribute to the development of a T-cell dependent allergic immune response.

  11. Recent Progress on Systems and Synthetic Biology Approaches to Engineer Fungi As Microbial Cell Factories.

    Science.gov (United States)

    Amores, Gerardo Ruiz; Guazzaroni, María-Eugenia; Arruda, Letícia Magalhães; Silva-Rocha, Rafael

    2016-04-01

    Filamentous fungi are remarkable organisms naturally specialized in deconstructing plant biomass and this feature has a tremendous potential for biofuel production from renewable sources. The past decades have been marked by a remarkable progress in the genetic engineering of fungi to generate industry-compatible strains needed for some biotech applications. In this sense, progress in this field has been marked by the utilization of high-throughput techniques to gain deep understanding of the molecular machinery controlling the physiology of these organisms, starting thus the Systems Biology era of fungi. Additionally, genetic engineering has been extensively applied to modify wellcharacterized promoters in order to construct new expression systems with enhanced performance under the conditions of interest. In this review, we discuss some aspects related to significant progress in the understating and engineering of fungi for biotechnological applications, with special focus on the construction of synthetic promoters and circuits in organisms relevant for industry. Different engineering approaches are shown, and their potential and limitations for the construction of complex synthetic circuits in these organisms are examined. Finally, we discuss the impact of engineered promoter architecture in the single-cell behavior of the system, an often-neglected relationship with a tremendous impact in the final performance of the process of interest. We expect to provide here some new directions to drive future research directed to the construction of high-performance, engineered fungal strains working as microbial cell factories.

  12. Industrial systems biology of Saccharomyces cerevisiae enables novel succinic acid cell factory.

    Directory of Open Access Journals (Sweden)

    José Manuel Otero

    Full Text Available Saccharomyces cerevisiae is the most well characterized eukaryote, the preferred microbial cell factory for the largest industrial biotechnology product (bioethanol, and a robust commerically compatible scaffold to be exploitted for diverse chemical production. Succinic acid is a highly sought after added-value chemical for which there is no native pre-disposition for production and accmulation in S. cerevisiae. The genome-scale metabolic network reconstruction of S. cerevisiae enabled in silico gene deletion predictions using an evolutionary programming method to couple biomass and succinate production. Glycine and serine, both essential amino acids required for biomass formation, are formed from both glycolytic and TCA cycle intermediates. Succinate formation results from the isocitrate lyase catalyzed conversion of isocitrate, and from the α-keto-glutarate dehydrogenase catalyzed conversion of α-keto-glutarate. Succinate is subsequently depleted by the succinate dehydrogenase complex. The metabolic engineering strategy identified included deletion of the primary succinate consuming reaction, Sdh3p, and interruption of glycolysis derived serine by deletion of 3-phosphoglycerate dehydrogenase, Ser3p/Ser33p. Pursuing these targets, a multi-gene deletion strain was constructed, and directed evolution with selection used to identify a succinate producing mutant. Physiological characterization coupled with integrated data analysis of transcriptome data in the metabolically engineered strain were used to identify 2(nd-round metabolic engineering targets. The resulting strain represents a 30-fold improvement in succinate titer, and a 43-fold improvement in succinate yield on biomass, with only a 2.8-fold decrease in the specific growth rate compared to the reference strain. Intuitive genetic targets for either over-expression or interruption of succinate producing or consuming pathways, respectively, do not lead to increased succinate. Rather, we

  13. Analyze of Impedance for Water Management in Proton Exchange Membrane Fuel Cells Using Factorial Design of (DoE) Methodology

    OpenAIRE

    Khaled Mammar; Abdelkader Chaker

    2014-01-01

    Electrochemical impedance spectroscopy (EIS) is a very powerful tool for exploitation as a rich source of Proton Exchange Membrane Fuel Cell (PEMFC) diagnostic information. A primary goal of this work is to develop a suitable PEMFC impedance model, which can be used to analyze flooding and drying of the fuel cell. For this one a novel optimization method based on Factorial Design methodology is used. It was applied to parametric analysis of electrochemical impedance Thus it is pos...

  14. Phagocytosis of Aspergillus fumigatus conidia by primary nasal epithelial cells in vitro

    Directory of Open Access Journals (Sweden)

    Khoufache Khaled

    2008-06-01

    Full Text Available Abstract Background Invasive aspergillosis, which is mainly caused by the fungus Aspergillus fumigatus, is an increasing problem in immunocompromised patients. Infection occurs by inhalation of airborne conidia, which are first encountered by airway epithelial cells. Internalization of these conidia into the epithelial cells could serve as a portal of entry for this pathogenic fungus. Results We used an in vitro model of primary cultures of human nasal epithelial cells (HNEC at an air-liquid interface. A. fumigatus conidia were compared to Penicillium chrysogenum conidia, a mould that is rarely responsible for invasive disease. Confocal microscopy, transmission electron microscopy, and anti-LAMP1 antibody labeling studies showed that conidia of both species were phagocytosed and trafficked into a late endosomal-lysosomal compartment as early as 4 h post-infection. In double immunolabeling experiments, the mean percentage of A. fumigatus conidia undergoing phagocytosis 4 h post-infection was 21.8 ± 4.5%. Using combined staining with a fluorescence brightener and propidium iodide, the mean rate of phagocytosis was 18.7 ± 9.3% and the killing rate 16.7 ± 7.5% for A. fumigatus after 8 h. The phagocytosis rate did not differ between the two fungal species for a given primary culture. No germination of the conidia was observed until 20 h of observation. Conclusion HNEC can phagocytose fungal conidia but killing of phagocytosed conidia is low, although the spores do not germinate. This phagocytosis does not seem to be specific to A. fumigatus. Other immune cells or mechanisms are required to kill A. fumigatus conidia and to avoid further invasion.

  15. Super Factories

    Indian Academy of Sciences (India)

    D G Hitlin

    2006-11-01

    Heavy-flavor physics, in particular and physics results from the factories, currently provides strong constraints on models of physics beyond the Standard Model. A new generation of colliders, Super Factories, with 50 to 100 times the luminosity of existing colliders, can, in a dialog with LHC and ILC, provide unique clarification of new physics phenomena seen at those machines.

  16. Ground-up circular Higgs Factory ring design and cell length optimization

    Science.gov (United States)

    Talman, Richard

    2017-02-01

    A “ground-up” Higgs Factory design methodology is described. For concreteness, numerical parameter choices are drawn primarily from CEPC, the Circular Electron Positron Collider. The goals are to find: (i) optimal parameters, (ii) improved understanding , (iii) a tentative lattice design. As illustration of the method, six chromaticity-corrected lattices, with cell lengths ranging from 45 m to 280 m, all with identical βy = 2 mm or βy = 10 mm intersection region optics, are designed and their properties compared. For simplicity only a single “toy ring,” circumference (76 km), with one interaction point, and a single beam energy (120 GeV) is considered. For the cell-length optimization a figure of merit FOM (essentially integrated luminosity) is maximized consistent with a dimensionless “fine tuning penalty function” or figure of demerit FOD not being allowed to exceed a conservatively chosen upper limit. The tentative recommendation from this investigation is that the optimal CEPC route is (except for obvious changes) to simply copy LEP: 80 m cell length and two-in-one single-ring operation. The main luminosity-increasing improvements are increased radius and power, top-off-full-energy-injection, noninterleaved sextupoles, more than 100 beam bunch operation, and improved intersection region design. Local chromaticity compensation (with its inevitable intense hard X-rays incident on the detectors) is found to be unnecessary. With these changes luminosity in excess of 1034cm‑2s‑1 is projected to be achievable.

  17. Physiological characteristics of the extreme thermophile Caldicellulosiruptor saccharolyticus: an efficient hydrogen cell factory

    Directory of Open Access Journals (Sweden)

    Zeidan Ahmad A

    2010-11-01

    Full Text Available Abstract Global concerns about climate changes and their association with the use of fossil fuels have accelerated research on biological fuel production. Biological hydrogen production from hemicellulose-containing waste is considered one of the promising avenues. A major economical issue for such a process, however, is the low substrate conversion efficiency. Interestingly, the extreme thermophilic bacterium Caldicellulosiruptor saccharolyticus can produce hydrogen from carbohydrate-rich substrates at yields close to the theoretical maximum of the dark fermentation process (i.e., 4 mol H2/mol hexose. The organism is able to ferment an array of mono-, di- and polysaccharides, and is relatively tolerant to high partial hydrogen pressures, making it a promising candidate for exploitation in a biohydrogen process. The behaviour of this Gram-positive bacterium bears all hallmarks of being adapted to an environment sparse in free sugars, which is further reflected in its low volumetric hydrogen productivity and low osmotolerance. These two properties need to be improved by at least a factor of 10 and 5, respectively, for a cost-effective industrial process. In this review, the physiological characteristics of C. saccharolyticus are analyzed in view of the requirements for an efficient hydrogen cell factory. A special emphasis is put on the tight regulation of hydrogen production in C. saccharolyticus by both redox and energy metabolism. Suggestions for strategies to overcome the current challenges facing the potential use of the organism in hydrogen production are also discussed.

  18. Apoptotic effect of demethoxyfumitremorgin C from marine fungus Aspergillus fumigatus on PC3 human prostate cancer cells.

    Science.gov (United States)

    Kim, Young-Sang; Kim, Se-Kwon; Park, Sun Joo

    2017-03-28

    Demethoxyfumitremorgin C, a secondary metabolite of the marine fungus, Aspergillus fumigatus, had been reported to demonstrate cytotoxic effect on mouse tsFT210 cells. However, no information is available regarding its functional mechanism and the chemo-sensitization effects on different kinds of human cancer cells. We found that treatment of demethoxyfumitremorgin C inhibited the cell viability of PC3 human advanced prostate cancer cells, induced apoptosis as determined by Annexin V/propidium iodide double staining, and decreased mitochondrial membrane potential. Demethoxyfumitremorgin C induced apoptosis was associated with downregulation of anti-apoptotic proteins: Ras, PI3K, Akt, Bcl-xL, and Bcl-2, and upregulation of pro-apoptotic Bax. Demethoxyfumitremorgin C activated caspase-3, -8, and -9, leading to PARP cleavage. Additionally, caspase inhibitors blocked demethoxyfumitremorgin C-induced apoptosis of PC3 cells. These results suggest that demethoxyfumitremorgin C from Aspergillus fumigatus inhibits the proliferation of PC3 human prostate cancer cells via the intrinsic (mitochondrial) and extrinsic pathway, followed by downstream events leading to apoptotic cell death. Demethoxyfumitremorgin C could therefore, serve as a useful agent to treat human advanced prostate cancer.

  19. Localization and function of calmodulin in live-cells of Aspergillus nidulans.

    Science.gov (United States)

    Chen, Shaochun; Song, Yiju; Cao, Jinling; Wang, Gang; Wei, Hua; Xu, Xushi; Lu, Ling

    2010-03-01

    Calmodulin (CaM) is a small, eukaryotic protein that reversibly binds Ca(2+). Study of CaM localization in genetically tractable organisms has yielded many insights into CaM function. Here, we described the dynamic localization of Aspergillus nidulans CaM (AnCaM) in live-cells by using recombination strains with homologous, single cross-over insertions at the target gene which placed the GFP fused copy under the inducible alcA promoter and the RFP-CaM integration under the native cam promoter. We found that the localization of CaM fusion was quite dynamic throughout the hypha and was concentrated to the active growing sites during germination, hyphal growth, cytokinesis and conidiation. The depletion of CaM by alcA promoter repression induced the explicit abnormalities of germlings with the swollen germ tubes. In addition, the position of highly concentrated GFP-CaM in the extreme apex seemed to determine the hyphal orientation. These data collectively suggest that CaM is constantly required for new hyphal growth. In contrast to this constant accumulation at the apex, GFP-CaM was only transiently localized at septum sites during cytokinesis. Notably, depletion of CaM caused the defect of septation with a completely blocked septum formation indicating that the transient CaM accumulation at the septum site is essential for septation. Moreover, the normal localization of CaM at a hyphal tip required the presence of the functional actin cytoskeleton and the motor protein KipA, which is indispensable for positioning Spitzenkörper. This is the first report of CaM localization and function in live-cells by the site-specific homologous integration in filamentous fungi.

  20. Dectin-1 agonist curdlan modulates innate immunity to Aspergillus fumigatus in human corneal epithelial cells

    Institute of Scientific and Technical Information of China (English)

    Cheng-Cheng; Zhu; Gui-Qiu; Zhao; Jing; Lin; Li-Ting; Hu; Qiang; Xu; Xu-Dong; Peng; Xue; Wang; Sheng; Qiu

    2015-01-01

    · AIM: To explore the immunomodulatory effects of curdlan on innate immune responses against Aspergillus fumigatus(A. fumigatus) in cultured human corneal epithelial cells(HCECs), and whether C-type lectin receptor Dectin-1 mediates the immunomodulatory effects of curdlan.·METHODS: The HCECs were stimulated by curdlan in different concentrations(50, 100, 200, 400 μg/m L) for various time. Then HCECs pretreated with or without laminarin(Dectin-1 blocker, 0.3 mg/m L) and curdlan were stimulated by A. fumigatus hyphae. The m RNA and protein production of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6) were determined by real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The protein level of Dectin-1 was measured by Western blot.· RESULTS: Curdlan stimulated m RNA expression of TNF-α and IL-6 in a dose and time dependent manner in HCECs. Curdlan pretreatment before A. fumigatus hyphae stimulation significantly enhanced the expression of TNF-α and IL-6 at m RNA and protein levels compared with A. fumigatus hyphae stimulation group(P <0.05).Both curdlan and A. fumigatus hyphae up-regulated Dectin-1 protein expression in HCECs, and Dectin-1expression was elevated to 1.5- to 2-fold by curdlan pretreatment followed hyphae stimulation. The Dectin-1blocker laminarin suppressed the m RNA expression and protein production of TNF-α and IL-6 induced by curdlan and hyphae(P <0.05).· CONCLUSION: These findings demonstrated that curdlan pretreatment enhanced the inflammatory response induced by A. fumigatus hyphae in HCECs.Dectin-1 is essential for the immunomodulatory effectsof curdlan. Curdlan may have high clinical application values in fungal keratitis treatment.

  1. New transposon tools tailored for metabolic engineering of Gram-negative microbial cell factories

    Directory of Open Access Journals (Sweden)

    Esteban eMartínez-García

    2014-10-01

    Full Text Available Re-programming microorganisms to modify their existing functions and/or to bestow bacteria with entirely new-to-Nature tasks have largely relied so far on specialized molecular biology tools. Such endeavors are not only relevant in the burgeoning metabolic engineering arena, but also instrumental to explore the functioning of complex regulatory networks from a fundamental point of view. À la carte modification of bacterial genomes thus calls for novel tools to make genetic manipulations easier. We propose the use of a series of new broad-host-range mini-Tn5 vectors, termed pBAMDs, for the delivery of gene(s into the chromosome of Gram-negative bacteria and for generating saturated mutagenesis libraries in gene function studies. These delivery vectors endow the user with the possibility of easy cloning and subsequent insertion of functional cargoes with three different antibiotic resistance markers (kanamycin, streptomycin, and gentamicin. After validating the pBAMD vectors in the environmental bacterium Pseudomonas putida KT2440, their use was also illustrated by inserting the entire poly(3-hydroxybutyrate (PHB synthesis pathway from Cupriavidus necator in the chromosome of a phosphotransacetylase mutant of Escherichia coli. PHB is a completely biodegradable polyester with a number of industrial applications that make it attractive as a potential replacement of oil-based plastics. The non-selective nature of chromosomal insertions of the biosynthetic genes was evidenced by a large landscape of PHB synthesis levels in independent clones. One clone was selected and further characterized as a microbial cell factory for PHB accumulation, and it achieved polymer accumulation levels comparable to those of a plasmid-bearing recombinant. Taken together, our results demonstrate that the new mini-Tn5 vectors can be used to confer interesting phenotypes in Gram-negative bacteria that would be very difficult to engineer through direct manipulation of the

  2. NIGERLYSINTM, HEMOLYSIN PRODUCED BY ASPERGILLUS NIGER, CAUSES LETHALITY OF PRIMARY RAT CORTICAL NEURONAL CELLS IN VITRO

    Science.gov (United States)

    Aspergillus niger produced a proteinaceous hemolysin, nigerlysinTM when incubated on sheep's blood agar at both 23° C and 37°C. Nigerlysin was purified from tryptic soy broth culture filtrate. Purified nigerlysin has a molecular weight of approximately 72 kDa, with an...

  3. Secondary metabolite production in Hypericum perforatum L. cell suspensions upon elicitation with fungal mycelia from Aspergillus flavus

    Directory of Open Access Journals (Sweden)

    Gadzovska-Simic Sonja

    2012-01-01

    Full Text Available We investigated the production of phenylpropanoids (phenolic compounds, flavanols, flavonols and anthocyanins and naphtodianthrones (hypericins in elicited Hypericum perforatum L. cell suspensions. To determine whether secondary metabolite production could be enhanced, Hypericum cell suspensions were exposed to mycelia extract from the fungus Aspergillus flavus. Elicited Hypericum cell suspension cultures displayed reduced growth and viability and a modification of secondary metabolites production. Anthocyanins were only stimulated in fungal-elicited cell suspensions. Secondary metabolite production in elicited Hypericum cells revealed an antagonism between the flavonoid/naphtodianthrone and anthocyanin pathways. The data suggest a modification of the channeling of the phenylpropanoid compounds. Together, these results represent useful data for monitoring the channeling in different secondary metabolite pathways during the scaled-up production of naphtodianthrones for medicinal uses.

  4. Analyze of Impedance for Water Management in Proton Exchange Membrane Fuel Cells Using Factorial Design of (DoE Methodology

    Directory of Open Access Journals (Sweden)

    Khaled Mammar

    2014-12-01

    Full Text Available Electrochemical impedance spectroscopy (EIS is a very powerful tool for exploitation as a rich source of Proton Exchange Membrane Fuel Cell (PEMFC diagnostic information. A primary goal of this work is to develop a suitable PEMFC impedance model, which can be used to analyze flooding and drying of the fuel cell. For this one a novel optimization method based on Factorial Design methodology is used. It was applied to parametric analysis of electrochemical impedance Thus it is possible to evaluate the relative importance of each parameter to the simulation accuracy. Furthermore this work presents an analysis of the PEMFC impedance behavior in the case of flooding and drying.

  5. beta-1,3-Glucan-Induced Host Phospholipase D Activation Is Involved in Aspergillus fumigatus Internalization into Type II Human Pneumocyte A549 Cells

    NARCIS (Netherlands)

    Han, Xuelin; Yu, Rentao; Zhen, Dongyu; Tao, Sha; Schmidt, Martina; Han, Li

    2011-01-01

    The internalization of Aspergillus fumigatus into lung epithelial cells is a process that depends on host cell actin dynamics. The host membrane phosphatidylcholine cleavage driven by phospholipase D (PLD) is closely related to cellular actin dynamics. However, little is known about the impact of PL

  6. Smart Factory

    DEFF Research Database (Denmark)

    Bilberg, Arne; Radziwon, Agnieszka; Grube Hansen, David

    2017-01-01

    , and to target their challenges and ensure sustainable growth and business in these enterprises. Therefore the focus of the Smart Factory project was to support the growth and sustainable development of the small and medium sized manufacturing industry in Denmark. The project focused on SMEs and how to improve...... their innovation and competitive advantage by focusing at their competences, strengths and opportunities. The project suggests innovative solutions and business models through collaboration and use of new technologies. In the Smart Factory, SMEs should be able to collaborate on new products, markets and production...... or supply chains in a creative organization. The Smart factory is supposed to be organic, meaning it is able to reconfigure and adjust to new projects and customers and has to be seen and taken as a model for inspiration to manufacturing businesses in general. The project takes into consideration Danish...

  7. Neutrino Factory

    CERN Document Server

    Bogomilov, M; Tsenov, R; Dracos, M; Bonesini, M; Palladino, V; Tortora, L; Mori, Y; Planche, T; Lagrange, J  B; Kuno, Y; Benedetto, E; Efthymiopoulos, I; Garoby, R; Gilardoini, S; Martini, M; Wildner, E; Prior, G; Blondel, A; Karadzhow, Y; Ellis, M; Kyberd, P; Bayes, R; Laing, A; Soler, F  J  P; Alekou, A; Apollonio, M; Aslaninejad, M; Bontoiu, C; Jenner, L  J; Kurup, A; Long, K; Pasternak, J; Zarrebini, A; Poslimski, J; Blackmore, V; Cobb, J; Tunnell, C; Andreopoulos, C; Bennett, J  R  J; Brooks, S; Caretta, O; Davenne, T; Densham, C; Edgecock, T  R; Fitton, M; Kelliher, D; Loveridge, P; McFarland, A; Machida, S; Prior, C; Rees, G; Rogers, C; Rooney, M; Thomason, J; Wilcox, D; Booth, C; Skoro, G; Back, J  J; Harrison, P; Berg, J  S; Fernow, R; Gallardo, J  C; Gupta, R; Kirk, H; Simos, N; Stratakis, D; Souchlas, N; Witte, H; Bross, A; Geer, S; Johnstone, C; Mokhov, N; Neuffer, D; Popovic, M; Strait, J; Striganov, S; Morfín, J  G; Wands, R; Snopok, P; Bogacz, S  A; Morozov, V; Roblin, Y; Cline, D; Ding, X; Bromberg, C; Hart, T; Abrams, R  J; Ankenbrandt, C  M; Beard, K  B; Cummings, M  A  C; Flanagan, G; Johnson, R  P; Roberts, T  J; Yoshikawa, C  Y; Graves, V  B; McDonald, K  T; Coney, L; Hanson, G

    2014-01-01

    The properties of the neutrino provide a unique window on physics beyond that described by the standard model. The study of subleading effects in neutrino oscillations, and the race to discover CP-invariance violation in the lepton sector, has begun with the recent discovery that $\\theta_{13} > 0$. The measured value of $\\theta_{13}$ is large, emphasizing the need for a facility at which the systematic uncertainties can be reduced to the percent level. The neutrino factory, in which intense neutrino beams are produced from the decay of muons, has been shown to outperform all realistic alternatives and to be capable of making measurements of the requisite precision. Its unique discovery potential arises from the fact that only at the neutrino factory is it practical to produce high-energy electron (anti)neutrino beams of the required intensity. This paper presents the conceptual design of the neutrino factory accelerator facility developed by the European Commission Framework Programme 7 EURO$\

  8. Aspergillus niger β-galactosidase production by yeast in a continuous high cell density reactor

    OpenAIRE

    2005-01-01

    The continuous production of extracellular heterologous beta-galactosidase by a recombinant flocculating Saccharomyces cerevisiae, expressing the lacA gene (coding for beta-galactosidase) of Aspergillus niger was investigated. A continuous operation was run in a 6.51 airlift bioreactor with a concentric draft tube using lactose as substrate. Data on the operation with semi-synthetic medium with 50 and 100 g/l initial lactose concentrations are presented. The best result for beta-galactosidase...

  9. Gliotoxin promotes Aspergillus fumigatus internalization into type II human pneumocyte A549 cells by inducing host phospholipase D activation.

    Science.gov (United States)

    Jia, Xiaodong; Chen, Fangyan; Pan, Weihua; Yu, Rentao; Tian, Shuguang; Han, Gaige; Fang, Haiqin; Wang, Shuo; Zhao, Jingya; Li, Xianping; Zheng, Dongyu; Tao, Sha; Liao, Wanqing; Han, Xuelin; Han, Li

    2014-06-01

    The internalization of Aspergillus fumigatus into lung epithelial cells is critical for the infection process in the host. Gliotoxin is the most potent toxin produced by A. fumigatus. However, its role in A. fumigatus internalization into the lung epithelial cells is still largely unknown. In the present study, the deletion of the gliP gene regulating the production of gliotoxin in A. fumigatus suppressed the internalization of conidia into the A549 lung epithelial cells, and this suppression could be rescued by the exogenous addition of gliotoxin. At lower concentrations, gliotoxin enhanced the internalization of the conidia of A. fumigatus into A549 cells; in contrast, it inhibited the phagocytosis of J774 macrophages in a dose-dependent manner. Under a concentration of 100 ng/ml, gliotoxin had no effect on A549 cell viability but attenuated ROS production in a dose-dependent manner. Gliotoxin significantly stimulated the phospholipase D activity in the A549 cells at a concentration of 50 ng/ml. This stimulation was blocked by the pretreatment of host cells with PLD1- but not PLD2-specific inhibitor. Morphological cell changes induced by gliotoxin were observed in the A549 cells accompanying with obvious actin cytoskeleton rearrangement and a moderate alteration of phospholipase D distribution. Our data indicated that gliotoxin might be responsible for modulating the A. fumigatus internalization into epithelial cells through phospholipase D1 activation and actin cytoskeleton rearrangement.

  10. Direct interaction studies between Aspergillus fumigatus and Human immune cells; what have we learned about pathogenicity and host immunity?

    Directory of Open Access Journals (Sweden)

    Charles Oliver Morton

    2012-12-01

    Full Text Available Invasive aspergillosis is a significant threat to health and is a major cause of mortality in immunocompromised individuals. Understanding the interaction between the fungus and the immune system is important in determining how the immunocompetent host remain disease free. Several studies examining the direct interaction between Aspergillus fumigatus and purified innate immune cells have been conducted to measure the responses of both the host cells and the pathogen. It has been revealed that innate immune cells have different modes of action ranging from effective fungal killing by neutrophils to the less aggressive response of dendritic cells. Natural-killer cells do not phagocytose the fungus unlike the other innate immune cells mentioned but appear to mediate their antifungal effect through the release of gamma interferon. Transcriptional analysis of A. fumigatus interacting with these cells has indicated that it can adapt to the harsh microenvironment of the phagosome and produces toxins, ribotoxin and gliotoxin, that can induce cell death in the majority of innate immune cells. These data point towards potential novel antifungal treatments including the use of innate immune cells as antifungal vaccines.

  11. Optimization of primary culture condition for mesenchymal stem cells derived from umbilical cord blood with factorial design.

    Science.gov (United States)

    Fan, Xiubo; Liu, Tianqing; Liu, Yang; Ma, Xuehu; Cui, Zhanfeng

    2009-01-01

    Mesenchymal stem cells (MSCs) can not only support the expansion of hematopoietic stem cells in vitro, but also alleviate complications and accelerate recovery of hematopoiesis during hematopoietic stem cell transplantation. However, it proved challenging to culture MSCs from umbilical cord blood (UCB) with a success rate of 20-30%. Many cell culture parameters contribute to this outcome and hence optimization of culture conditions is critical to increase the probability of success. In this work, fractional factorial design was applied to study the effect of cell inoculated density, combination and dose of cytokines, and presence of serum and stromal cells. The cultured UCB-MSC-like cells were characterized by flow cytometry and their multilineage differentiation potentials were tested. The optimal protocol was identified achieving above 90% successful outcome: 2 x 10(6) cells/mL mononuclear cells inoculated in Iscove's modified Dulbecco's medium supplied with 10% FBS, 15 ng/mL IL-3, and 5 ng/mL Granulocyte-macrophage colony-stimulating factor (GM-CSF). Moreover, the UCB-MSC-like cells expressed MSC surface markers of CD13, CD29, CD105, CD166, and CD44 positively, and CD34, CD45, and human leukocyte antigens-DR (HLA-DR) negatively. Meanwhile, these cells could differentiate into osteoblasts, chondrocytes, and adipocytes similarly to MSCs derived from bone marrow. In conclusion, we have developed an efficient protocol for the primary culture of UCB-MSCs by adding suitable cytokines into the culture system.

  12. Reshuffling of Aspergillus fumigatus cell wall components chitin and β-glucan under the influence of caspofungin or nikkomycin Z alone or in combination

    NARCIS (Netherlands)

    P.E.B. Verwer (Patricia); H.M. van Duijn (Miranda); M. Tavakol (Mehri); I.A.J.M. Bakker-Woudenberg (Irma); W.W.J. van de Sande (Wendy)

    2012-01-01

    textabstractChitin and β-glucan are major cell wall components of Aspergillus spp. We investigated the antifungal activity of chitin synthesis inhibitors nikkomycin Z, polyoxin D, flufenoxuron, lufenuron, and teflubenzuron, alone and combined with the β-glucan synthesis inhibitor caspofungin. Only n

  13. Activation of vitamin D regulates response of human bronchial epithelial cells to Aspergillus fumigatus in an autocrine fashion.

    Science.gov (United States)

    Li, Pei; Wu, Ting; Su, Xin; Shi, Yi

    2015-01-01

    Aspergillus fumigatus (A. fumigatus) is one of the most common fungi to cause diseases in humans. Recent evidence has demonstrated that airway epithelial cells play an important role in combating A. fumigatus through inflammatory responses. Human airway epithelial cells have been proven to synthesize the active vitamin D, which plays a key role in regulating inflammation. The present study was conducted to investigate the impact of A. fumigatus infection on the activation of vitamin D and the role of vitamin D activation in A. fumigatus-elicited antifungal immunity in normal human airway epithelial cells. We found that A. fumigatus swollen conidia (SC) induced the expression of 1α-hydroxylase, the enzyme catalyzing the synthesis of active vitamin D, and vitamin D receptor (VDR) in 16HBE cells and led to increased local generation of active vitamin D. Locally activated vitamin D amplified SC-induced expression of antimicrobial peptides in 16HBE cells but attenuated SC-induced production of cytokines in an autocrine fashion. Furthermore, we identified β-glucan, the major A. fumigatus cell wall component, as the causative agent for upregulation of 1α-hydroxylase and VDR in 16HBE cells. Therefore, activation of vitamin D is inducible and provides a bidirectional regulation of the responses to A. fumigatus in 16HBE cells.

  14. Cell factory applications of the yeast Kluyveromyces marxianus for the biotechnological production of natural flavour and fragrance molecules.

    Science.gov (United States)

    Morrissey, John P; Etschmann, Maria M W; Schrader, Jens; de Billerbeck, Gustavo M

    2015-01-01

    Kluyveromyces marxianus is emerging as a new platform organism for the production of flavour and fragrance (F&F) compounds. This food-grade yeast has advantageous traits, such as thermotolerance and rapid growth, that make it attractive for cell factory applications. The major impediment to its development has been limited fundamental knowledge of its genetics and physiology, but this is rapidly changing. K. marxianus produces a wide array of volatile molecules and contributes to the flavour of a range of different fermented beverages. Advantage is now being taken of this to develop strains for the production of metabolites such as 2-phenylethanol and ethyl acetate. Strains that were selected from initial screens were used to optimize processes for production of these F&F molecules. Most developments have focused on optimizing growth conditions and the fermentation process, including product removal, with future advancement likely to involve development of new strains through the application of evolutionary or rational engineering strategies. This is being facilitated by new genomic and molecular tools. Furthermore, synthetic biology offers a route to introduce new biosynthetic pathways into this yeast for F&F production. Consumer demand for biologically-synthesized molecules for use in foods and other products creates an opportunity to exploit the unique potential of K. marxianus for this cell factory application.

  15. Proteases from Aspergillus fumigatus induce release of proinflammatory cytokines and cell detachment in airway epithelial cell lines

    NARCIS (Netherlands)

    Tomee, JFC; Hiemstra, PS; Kauffman, HF

    1997-01-01

    Aspergillus fumigatus is a pathogen causing; diverse respiratory disorders, Several studies have suggested that fungal proteases may play a role in the pathogenicity of fungi, Since the airways are the most common route for entry of. A, fumigatus, this study focused on the ability of fungal protease

  16. Human tumor cells induce angiogenesis through positive feedback between CD147 and insulin-like growth factor-I.

    Directory of Open Access Journals (Sweden)

    Yanke Chen

    Full Text Available Tumor angiogenesis is a complex process based upon a sequence of interactions between tumor cells and endothelial cells. Previous studies have shown that CD147 was correlated with tumor angiogenesis through increasing tumor cell secretion of vascular endothelial growth factor (VEGF and matrix metalloproteinases (MMPs. In this study, we made a three-dimensional (3D tumor angiogenesis model using a co-culture system of human hepatocellular carcinoma cells SMMC-7721 and humanumbilical vein endothelial cells (HUVECs in vitro. We found that CD147-expressing cancer cells could promote HUVECs to form net-like structures resembling the neo-vasculature, whereas the ability of proliferation, migration and tube formation of HUVECs was significantly decreased in tumor conditioned medium (TCM of SMMC-7721 cells transfected with specific CD147-siRNA. Furthermore, by assaying the change of pro-angiogenic factors in TCM, we found that the inhibition of CD147 expression led to significant decrease of VEGF and insulin-like growth factor-I (IGF-I secretion. Interestingly, we also found that IGF-I up-regulated the expression of CD147 in both tumor cells and HUVECs. These findings suggest that there is a positive feedback between CD147 and IGF-I at the tumor-endothelial interface and CD147 initiates the formation of an angiogenesis niche.

  17. On-line study of growth kinetics of single hyphae of Aspergillus oryzae in a flow-through cell

    DEFF Research Database (Denmark)

    Christiansen, Torben; Spohr, Anders Bendsen; Nielsen, Jens Bredal

    1999-01-01

    Using image analysis the growth kinetics of the single hyphae of the filamentous fungus Aspergillus oryzae has been determined on-line in a flow-through cell at different glucose concentrations in the range from 26 mg L-1 to 20 g L-1. The tip extension rate of the individual hyphae can be described...... with saturation type kinetics with respect to the length of the hyphae. The maximum tip extension rate is constant for all hyphae measured at the same glucose concentration, whereas the saturation constant for the hyphae varies significantly between the hyphae even within the same hyphal element. When apical...... branching occurs, it is observed that the tip extension rate decreases temporarily. The number of branches formed on a hypha is proportional to the length of the hypha that exceeds a certain minimum length required to support the growth of a new branch. The observed kinetics has been used to simulate...

  18. Lobar Collapse and Obliteration of Air Bronchogram Allowing Early Diagnosis of Endobronchial Aspergillus Infection following Hematopoietic Stem Cell Transplantation

    Directory of Open Access Journals (Sweden)

    Elizabeth Thompson

    2014-01-01

    Full Text Available Endobronchial fungal infection (EBFI is notoriously difficult to diagnose early since it may present few systemic features and does not cause characteristic parenchymal lesions on lung CT scanning. We report a 9-year-old girl who suffered extended neutropenia following graft failure after haematopoietic stem cell transplantation (HSCT for severe aplastic anaemia. CT scan prior to retransplantation was normal despite persistent cough but lobar collapse was shown on repeat scan 16 days later. The probable diagnosis of EBFI (later proven on bronchoscopy was only suspected when subsequent chest X-ray (CXR demonstrated lack of an air bronchogram in the partially collapsed lung. Early radiological suspicion resulted in multiagent antifungal therapy followed by delayed lobectomy, and led to this being the first reported case of Aspergillus EBFI not to result in respiratory failure.

  19. Alginate Oligosaccharides Inhibit Fungal Cell Growth and Potentiate the Activity of Antifungals against Candida and Aspergillus spp

    Science.gov (United States)

    Tøndervik, Anne; Sletta, Håvard; Klinkenberg, Geir; Emanuel, Charlotte; Powell, Lydia C.; Pritchard, Manon F.; Khan, Saira; Craine, Kieron M.; Onsøyen, Edvar; Rye, Phil D.; Wright, Chris; Thomas, David W.; Hill, Katja E.

    2014-01-01

    The oligosaccharide OligoG, an alginate derived from seaweed, has been shown to have anti-bacterial and anti-biofilm properties and potentiates the activity of selected antibiotics against multi-drug resistant bacteria. The ability of OligoG to perturb fungal growth and potentiate conventional antifungal agents was evaluated using a range of pathogenic fungal strains. Candida (n = 11) and Aspergillus (n = 3) spp. were tested using germ tube assays, LIVE/DEAD staining, scanning electron microscopy (SEM), atomic force microscopy (AFM) and high-throughput minimum inhibition concentration assays (MICs). In general, the strains tested showed a significant dose-dependent reduction in cell growth at ≥6% OligoG as measured by optical density (OD600; P0.5%) also showed a significant inhibitory effect on hyphal growth in germ tube assays, although strain-dependent variations in efficacy were observed (P<0.05). SEM and AFM both showed that OligoG (≥2%) markedly disrupted fungal biofilm formation, both alone, and in combination with fluconazole. Cell surface roughness was also significantly increased by the combination treatment (P<0.001). High-throughput robotic MIC screening demonstrated the potentiating effects of OligoG (2, 6, 10%) with nystatin, amphotericin B, fluconazole, miconazole, voriconazole or terbinafine with the test strains. Potentiating effects were observed for the Aspergillus strains with all six antifungal agents, with an up to 16-fold (nystatin) reduction in MIC. Similarly, all the Candida spp. showed potentiation with nystatin (up to 16-fold) and fluconazole (up to 8-fold). These findings demonstrate the antifungal properties of OligoG and suggest a potential role in the management of fungal infections and possible reduction of antifungal toxicity. PMID:25409186

  20. Triggering of toll-like receptors 2 and 4 by Aspergillus fumigatus conidia in immortalized human corneal epithelial cells to induce inflammatory cytokines

    Institute of Scientific and Technical Information of China (English)

    ZHAO Jie; WU Xin-yi

    2008-01-01

    Background Cornea epithelial cells play early and crucial roles in the initiation of ocular surface responses to pathogens.Participation of toll-like receptor(TLR)2 and TLR4,which are major forms of fungi receptors,may be involved in Aspergillus fumigatus induced immune responses.The obiective of the present study was to examine whether inactive Aspergillus fumigatus conidia induce NF-κB activation and production of proinflammaory cytokines,and whether the expression of TLR2 and TLR4 were amplified by conidia in cultured immortalized human corneal epithelial cells (THCEs).This may contribute to our knowledge of the mechanism by which the host cornea can successfully defend against invasive fungi.Methods Aspergillus fumigatus conidia were used to challenge THCE cells.THCE cells were harvested after 0.5,1,2or 4 hours incubation.Real-time quantitative PCR was performed to determine the expression of TLR2,TLR4,TNF-α and IL-8.Western blotting was performed to determine the expression of NF-κB.Enzyme-linked immunosorbent assay (EUSA)was performed to determine the expression of TNF-α and IL-8.And the release of TNF-α and IL-8 in the cell supematant were also assessed by ELISA with or without pretreatment with TLR2 and TLR4 neutralizing antibodies.Results Aspergillus fumigatus conidia elicited the expression of TLR2,TLR4,TNF-α and IL-8 mRNA in THCEs.Exposure of THCE cells to Aspergillus fumigatus conidia resulted in NF-κB activation,which increased at 30 minutes (increased from 11.35±2.74 in the controls to 19.12±3.48,P<0.05)and thereafter increased steadily up to 4 hours after challenge(P<0.01).Concomitant with NF-κB acfivation,secretion of TNF-α and IL-8 in conidia-challenged cells was increased in a time-dependent manner.Incubation of THCE cells with TLR2 antibody or TLR4 antibody before conidia challenge resulted in jnhibifion of conidia-induced TNF-α and IL-8 secretion(P<0.05),TLR2 antibody and TLR4 antibody together significantly increased

  1. Aspergillus oryzae lectin induces anaphylactoid oedema and mast cell activation through its interaction with fucose of mast cell-bound non-specific IgE.

    Science.gov (United States)

    Yamaki, K; Yoshino, S

    2011-11-01

    We investigated whether Aspergillus oryzae lectin (AOL), a fucose-specific lectin, induces anaphylactoid reactions and mast cell activation. The injection of AOL into footpads of mice produced a dose-related acute paw oedema. The AOL-induced oedema was attenuated by predose of histamine H1 receptor blocker or pretreatment of the lectin with fucose before injection and was not observed in SCID and mast cell-deficient WBB6F1-W/Wv mice. These results suggested that the AOL-induced anaphylactoid reaction was mediated by histamine released from mast cells. In addition, the activation of mast cells was seemed to be induced by the crosslinking of IgE on the cell surface following the binding of AOL to fucose residues in IgE. Consistent with the in vivo results, AOL induced the degranulation of the rat mast cell line RBL2H3 sensitized with monoclonal IgE. As AOL induced the increase in intracellular Ca(2+) concentration of IgE-sensitized RBL2H3 cells as well as antigen stimulation, AOL could input signals from FcεRI. The degranulation of IgE-sensitized RBL2H3 cells by AOL was diminished by pretreatment of AOL with fucose. Defucosylated IgE did not induce degranulation of RBL2H3 cells in response to AOL stimulation, in spite of its ability to induce degranulation by antigen stimulation as intact IgE. These results indicated that AOL bound to fucose residue of IgE causing antigen-independent IgE-mediated mast cell activation and anaphylactoid reactions in vitro and in vivo, respectively. AOL bound to human IgE as well as to mouse IgE, suggesting the possible implication of AOL in the allergic response to Aspergillus oryzae in humans.

  2. Flow-cytometry and cell sorting: an efficient approach to investigate productivity and cell physiology in mammalian cell factories.

    Science.gov (United States)

    Kumar, Niraj; Borth, Nicole

    2012-03-01

    The performance of cell lines used for the production of biotherapeutic proteins typically depends on the number of cells in culture, their specific growth rate, their viability and the cell specific productivity (qP). Therefore both cell line development and process development are trying to (a) improve cell proliferation to reduce lag-phase and achieve high number of cells; (b) delay cell death to prolong the production phase and improve culture longevity; (c) and finally, increase qP. All of these factors, when combined in an optimised process, concur to increase the final titre and yield of the recombinant protein. As cellular performance is at the centre of any improvement, analysis methods that enable the characterisation of individual cells in their entirety can help in identifying cell types and culture conditions that perform exceptionally well. This observation of cells and their complexity is reflected by the term "cytomics" and flow cytometry is one of the methods used for this purpose. With its ability to analyse the distribution of physiological properties within a population and to isolate rare outliers with exceptional properties, flow cytometry ideally complements other methods used for optimisation, including media design and cell engineering. In the present review we describe approaches that could be used, directly or indirectly, to analyse and sort cellular phenotypes characterised by improved growth behaviour, reduced cell death or high qP and outline their potential use for cell line and process optimisation.

  3. Skeletal unloading causes resistance of osteoprogenitor cells to parathyroid hormone and to insulin-like growth factor-I

    Science.gov (United States)

    Kostenuik, P. J.; Harris, J.; Halloran, B. P.; Turner, R. T.; Morey-Holton, E. R.; Bikle, D. D.

    1999-01-01

    Skeletal unloading decreases bone formation and osteoblast number in vivo and decreases the number and proliferation of bone marrow osteoprogenitor (BMOp) cells in vitro. We tested the ability of parathyroid hormone (PTH) to stimulate BMOp cells in vivo by treating Sprague Dawley rats (n = 32) with intermittent PTH(1-34) (1 h/day at 8 microg/100 g of body weight), or with vehicle via osmotic minipumps during 7 days of normal weight bearing or hind limb unloading. Marrow cells were flushed from the femur and cultured at the same initial density for up to 21 days. PTH treatment of normally loaded rats caused a 2.5-fold increase in the number of BMOp cells, with similar increases in alkaline phosphatase (ALP) activity and mineralization, compared with cultures from vehicle-treated rats. PTH treatment of hind limb unloaded rats failed to stimulate BMOp cell number, ALP activity, or mineralization. Hind limb unloading had no significant effect on PTH receptor mRNA or protein levels in the tibia. Direct in vitro PTH challenge of BMOp cells isolated from normally loaded bone failed to stimulate their proliferation and inhibited their differentiation, suggesting that the in vivo anabolic effect of intermittent PTH on BMOp cells was mediated indirectly by a PTH-induced factor. We hypothesize that this factor is insulin-like growth factor-I (IGF-I), which stimulated the in vitro proliferation and differentiation of BMOp cells isolated from normally loaded bone, but not from unloaded bone. These results suggest that IGF-I mediates the ability of PTH to stimulate BMOp cell proliferation in normally loaded bone, and that BMOp cells in unloaded bone are resistant to the anabolic effect of intermittent PTH therapy due to their resistance to IGF-I.

  4. Molecular tools and protocols for engineering the acid-tolerant yeast Zygosaccharomyces bailii as a potential cell factory.

    Science.gov (United States)

    Branduardi, Paola; Dato, Laura; Porro, Danilo

    2014-01-01

    Microorganisms offer a tremendous potential as cell factories, and they are indeed used by humans for centuries for biotransformations. Among them, yeasts combine the advantage of unicellular state with a eukaryotic organization, and, in the era of biorefineries, their biodiversity can offer solutions to specific process constraints. Zygosaccharomyces bailii, an ascomycetales budding yeast, is widely known for its peculiar tolerance to various stresses, among which are organic acids. Despite the possibility to apply with this yeast some of the molecular tools and protocols routinely used to manipulate Saccharomyces cerevisiae, adjustments and optimizations are necessary. Here, we describe in detail protocols for transformation, for target gene disruption or gene integration, and for designing episomal expression plasmids helpful for developing and further studying the yeast Z. bailii.

  5. Identification of potentially safe promising fungal cell factories for the production of polyketide natural food colorants using chemotaxonomic rationale

    DEFF Research Database (Denmark)

    Mapari, Sameer Shamsuddin; Meyer, Anne S.; Thrane, Ulf

    2009-01-01

    chemical diversity and biodiversity of ascomycetous fungi. This rationale could be very handy for the selection of potentially safe fungal cell factories not only for polyketide pigments but also for the other industrially important polyketides; the molecular and genetic basis for the biosynthesis of which...... provide readily available additional and/or alternative sources of natural colorants that are independent of agro-climatic conditions. With an appropriately selected fungus; using in particular chemotaxonomy as a guide, the fungal natural colorants could be produced in high yields by using the optimized...... cultivation technology. This approach could secure efficient production of pigments avoiding use of genetic manipulation. Results: Polyketide pigment producing ascomycetous fungi were evaluated for their potential as production organisms based on a priori knowledge on species-specific pigment and potential...

  6. Characterization of a Solid Oxide Fuel Cell Gas Turbine Hybrid System Based on a Factorial Design of Experiments Using Hardware Simulation

    Energy Technology Data Exchange (ETDEWEB)

    Restrepo, Bernardo; Banta, Larry E; Tucker, David

    2012-10-01

    A full factorial experimental design and a replicated fractional factorial design were carried out using the Hybrid Performance (HyPer) project facility installed at the National Energy Technology Laboratory (NETL), U.S. Department of Energy to simulate gasifer/fuel cell/turbine hybrid power systems. The HyPer facility uses hardware in the loop (HIL) technology that couples a modified recuperated gas turbine cycle with hardware driven by a solid oxide fuel cell model. A 34 full factorial design (FFD) was selected to study the effects of four factors: cold-air, hot-air, bleed-air bypass valves, and the electric load on different parameters such as cathode and turbine inlet temperatures, pressure and mass flow. The results obtained, compared with former results where the experiments were made using one-factor-at-a-time (OFAT), show that no strong interactions between the factors are present in the different parameters of the system. This work also presents a fractional factorial design (ffd) 34-2 in order to analyze replication of the experiments. In addition, a new envelope is described based on the results of the design of experiments (DoE), compared with OFAT experiments, and analyzed in an off-design integrated fuel cell/gas turbine framework. This paper describes the methodology, strategy, and results of these experiments that bring new knowledge concerning the operating state space for this kind of power generation system.

  7. Human Cortical Neural Stem Cells Expressing Insulin-Like Growth Factor-I: A Novel Cellular Therapy for Alzheimer's Disease.

    Science.gov (United States)

    McGinley, Lisa M; Sims, Erika; Lunn, J Simon; Kashlan, Osama N; Chen, Kevin S; Bruno, Elizabeth S; Pacut, Crystal M; Hazel, Tom; Johe, Karl; Sakowski, Stacey A; Feldman, Eva L

    2016-03-01

    Alzheimer's disease (AD) is the most prevalent age-related neurodegenerative disorder and a leading cause of dementia. Current treatment fails to modify underlying disease pathologies and very little progress has been made to develop effective drug treatments. Cellular therapies impact disease by multiple mechanisms, providing increased efficacy compared with traditional single-target approaches. In amyotrophic lateral sclerosis, we have shown that transplanted spinal neural stem cells (NSCs) integrate into the spinal cord, form synapses with the host, improve inflammation, and reduce disease-associated pathologies. Our current goal is to develop a similar "best in class" cellular therapy for AD. Here, we characterize a novel human cortex-derived NSC line modified to express insulin-like growth factor-I (IGF-I), HK532-IGF-I. Because IGF-I promotes neurogenesis and synaptogenesis in vivo, this enhanced NSC line offers additional environmental enrichment, enhanced neuroprotection, and a multifaceted approach to treating complex AD pathologies. We show that autocrine IGF-I production does not impact the cell secretome or normal cellular functions, including proliferation, migration, or maintenance of progenitor status. However, HK532-IGF-I cells preferentially differentiate into gamma-aminobutyric acid-ergic neurons, a subtype dysregulated in AD; produce increased vascular endothelial growth factor levels; and display an increased neuroprotective capacity in vitro. We also demonstrate that HK532-IGF-I cells survive peri-hippocampal transplantation in a murine AD model and exhibit long-term persistence in targeted brain areas. In conclusion, we believe that harnessing the benefits of cellular and IGF-I therapies together will provide the optimal therapeutic benefit to patients, and our findings support further preclinical development of HK532-IGF-I cells into a disease-modifying intervention for AD.

  8. Bacillus subtilis and Escherichia coli essential genes and minimal cell factories after one decade of genome engineering.

    Science.gov (United States)

    Juhas, Mario; Reuß, Daniel R; Zhu, Bingyao; Commichau, Fabian M

    2014-11-01

    Investigation of essential genes, besides contributing to understanding the fundamental principles of life, has numerous practical applications. Essential genes can be exploited as building blocks of a tightly controlled cell 'chassis'. Bacillus subtilis and Escherichia coli K-12 are both well-characterized model bacteria used as hosts for a plethora of biotechnological applications. Determination of the essential genes that constitute the B. subtilis and E. coli minimal genomes is therefore of the highest importance. Recent advances have led to the modification of the original B. subtilis and E. coli essential gene sets identified 10 years ago. Furthermore, significant progress has been made in the area of genome minimization of both model bacteria. This review provides an update, with particular emphasis on the current essential gene sets and their comparison with the original gene sets identified 10 years ago. Special attention is focused on the genome reduction analyses in B. subtilis and E. coli and the construction of minimal cell factories for industrial applications.

  9. The temporal dynamics of differential gene expression in Aspergillus fumigatus interacting with human immature dendritic cells in vitro.

    LENUS (Irish Health Repository)

    Morton, Charles O

    2011-01-01

    Dendritic cells (DC) are the most important antigen presenting cells and play a pivotal role in host immunity to infectious agents by acting as a bridge between the innate and adaptive immune systems. Monocyte-derived immature DCs (iDC) were infected with viable resting conidia of Aspergillus fumigatus (Af293) for 12 hours at an MOI of 5; cells were sampled every three hours. RNA was extracted from both organisms at each time point and hybridised to microarrays. iDC cell death increased at 6 h in the presence of A. fumigatus which coincided with fungal germ tube emergence; >80% of conidia were associated with iDC. Over the time course A. fumigatus differentially regulated 210 genes, FunCat analysis indicated significant up-regulation of genes involved in fermentation, drug transport, pathogenesis and response to oxidative stress. Genes related to cytotoxicity were differentially regulated but the gliotoxin biosynthesis genes were down regulated over the time course, while Aspf1 was up-regulated at 9 h and 12 h. There was an up-regulation of genes in the subtelomeric regions of the genome as the interaction progressed. The genes up-regulated by iDC in the presence of A. fumigatus indicated that they were producing a pro-inflammatory response which was consistent with previous transcriptome studies of iDC interacting with A. fumigatus germ tubes. This study shows that A. fumigatus adapts to phagocytosis by iDCs by utilising genes that allow it to survive the interaction rather than just up-regulation of specific virulence genes.

  10. The temporal dynamics of differential gene expression in Aspergillus fumigatus interacting with human immature dendritic cells in vitro.

    Directory of Open Access Journals (Sweden)

    Charles O Morton

    Full Text Available Dendritic cells (DC are the most important antigen presenting cells and play a pivotal role in host immunity to infectious agents by acting as a bridge between the innate and adaptive immune systems. Monocyte-derived immature DCs (iDC were infected with viable resting conidia of Aspergillus fumigatus (Af293 for 12 hours at an MOI of 5; cells were sampled every three hours. RNA was extracted from both organisms at each time point and hybridised to microarrays. iDC cell death increased at 6 h in the presence of A. fumigatus which coincided with fungal germ tube emergence; >80% of conidia were associated with iDC. Over the time course A. fumigatus differentially regulated 210 genes, FunCat analysis indicated significant up-regulation of genes involved in fermentation, drug transport, pathogenesis and response to oxidative stress. Genes related to cytotoxicity were differentially regulated but the gliotoxin biosynthesis genes were down regulated over the time course, while Aspf1 was up-regulated at 9 h and 12 h. There was an up-regulation of genes in the subtelomeric regions of the genome as the interaction progressed. The genes up-regulated by iDC in the presence of A. fumigatus indicated that they were producing a pro-inflammatory response which was consistent with previous transcriptome studies of iDC interacting with A. fumigatus germ tubes. This study shows that A. fumigatus adapts to phagocytosis by iDCs by utilising genes that allow it to survive the interaction rather than just up-regulation of specific virulence genes.

  11. The cell factory approach toward biotechnological production of high-value chitosan oligomers and their derivatives: an update.

    Science.gov (United States)

    Naqvi, Shoa; Moerschbacher, Bruno M

    2017-02-01

    Chitin is one of the most abundant renewable resources, and chitosans, the partially deacetylated derivatives of chitin, are among the most promising functional biopolymers, with superior material properties and versatile biological functionalities. Elucidating molecular structure-function relationships and cellular modes of action of chitosans, however, it is challenging due to the micro-heterogeneity and structural complexity of polysaccharides. Lately, it has become apparent that many of the biological activities of chitosan polymers, such as in agricultural plant disease protection or in mediating scar-free wound healing, may be attributed to oligomeric break-down products generated by the action of chitosanolytic hydrolases present in the target tissues, such as human chitotriosidase. Consequently, the focus of current research is shifting toward chitosan oligomers so that the availability of well-defined chitosan oligosaccharides (COS) becomes a bottleneck. Well-known ways of producing COS use physical and/or chemical means for the partial depolymerization of chitosan polymers, typically leading to broad mixtures of COS varying in their degrees of polymerization (DP) and acetylation (DA), and with more or less random patterns of acetylation (PAs). Even after chromatographic separation according to DP and DA, such mixtures are of limited value to elucidate structure-function relationships and modes of action. More recently, enzymatic means using chitinases and/or chitosanases, and sometimes chitin deacetylases, have been proposed as these can be more tightly controlled and yield slightly better defined mixtures of COS. An alternative would be chemical synthesis of COS which in principle would allow for full structural control, but protocols for it are lengthy, costly, and not yet well developed, and yields are low. Synthetic biology now allows to develop today's in vitro bio-refinery approaches into in vivo cell factory approaches for the biotechnological

  12. Identification of potentially safe promising fungal cell factories for the production of polyketide natural food colorants using chemotaxonomic rationale

    Directory of Open Access Journals (Sweden)

    Frisvad Jens C

    2009-04-01

    of chemotaxonomic tools and a priori knowledge of fungal extrolites is a rational approach towards selection of fungal polyketide pigment producers considering the enormous chemical diversity and biodiversity of ascomycetous fungi. This rationale could be very handy for the selection of potentially safe fungal cell factories not only for polyketide pigments but also for the other industrially important polyketides; the molecular and genetic basis for the biosynthesis of which has not yet been examined in detail. In addition, 4 out of the 10 chemotaxonomically selected promising Penicillium strains were shown to produce extracellular pigments in the liquid media using a solid support indicating future cell factory possibilities for polyketide natural food colorants.

  13. Highly efficient biodiesel production by a whole-cell biocatalyst employing a system with high lipase expression in Aspergillus oryzae.

    Science.gov (United States)

    Takaya, Tomohiro; Koda, Risa; Adachi, Daisuke; Nakashima, Kazunori; Wada, Junpei; Bogaki, Takayuki; Ogino, Chiaki; Kondo, Akihiko

    2011-05-01

    In the present study, a system with high lipase expression in Aspergillus oryzae was developed using an improved enolase promoter (P-enoA124) and the 5' untranslated region of a heat-shock protein (Hsp-UTR). P-enoA142 enhanced the transcriptional level of a heterologous lipase gene and Hsp-UTR improved its translational efficiency. Fusarium heterosporum lipase (FHL) was inserted into a pSENSU-FHL expression vector harboring P-enoA142 and Hsp-UTR and was transformed into an A. oryzae NS4 strain. Transformants possessing pSENSU-FHL in single (pSENSU-FHL#1) and double copies (pSENSU-FHL#2) were selected to evaluate the lipase activity of the whole-cell biocatalyst. The two strains, pSENSU-FHL#1 and #2, showed excellent lipase activity in hydrolysis compared with the strain transformed with conventional expression vector pNAN8142-FHL. Furthermore, by using pSENSU-FHL#2, methanolysis could proceed much more effectively without deactivation, which allowed a swift addition of methanol to the reaction mixture, thereby reducing reaction time.

  14. Early expression of mannose-binding lectin 2 during Aspergillus fumigatus infection in human corneal epithelial cells

    Institute of Scientific and Technical Information of China (English)

    Cheng-Ye; Che; Jing-Fen; Zhang; Ji-Eun; Lee; Jing; Lin; Li-Ting; Hu; Nan; Jiang; Qian; Wang; Qiang; Xu; Gui-Qiu; Zhao

    2015-01-01

    AIM: To evaluate the early expression of mannose-binding lectin 2(MBL2) in human corneal epithelial cells(HCECs) infected by Aspergillus fumigatus(AF).METHODS: HCECs cultured in vitro with AF antigens and sampled at 0, 0.5, 1, 2, 4, 6 and 8h. The expression of MBL2 m RNA was evaluated by semiquantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression of MBL2 protein in supernatant fluid was shown by enzyme linked immunosorbent assay(ELISA). MBL2 protein in HCECs was detected by immunocytochemistry at 0 and 24 h.RESULTS: MBL2 m RNA and protein are expressed in normal HCECs. The expression of MBL2 m RNA and protein in supernatant fluid begin to increase after being stimulated with AF antigens. The most significantly peak of MBL2 m RNA is in 2h. The protein of MBL2 in supernatant fluid decrease gradually after 0.5h. The protein in HCECs expression increase after stimulation of24 h.· CONCLUSION: MBL2 receptor expressed in normal HCECs in vitro. The stimulation by AF antigens can increase the early expression of it.

  15. Highly efficient biodiesel production by a whole-cell biocatalyst employing a system with high lipase expression in Aspergillus oryzae

    Energy Technology Data Exchange (ETDEWEB)

    Takaya, Tomohiro; Koda, Risa; Adachi, Daisuke; Ogino, Chiaki; Kondo, Akihiko [Kobe Univ. (Japan). Dept. of Chemical Science and Engineering; Nakashima, Kazunori [Kobe Univ. (Japan). Organization of Advanced Science and Technology; Wada, Junpei; Bogaki, Takayuki [Ozeki Co., Nishinomiya-shi, Hyogo (Japan)

    2011-05-15

    In the present study, a system with high lipase expression in Aspergillus oryzae was developed using an improved enolase promoter (P-enoA124) and the 5{sup '} untranslated region of a heat-shock protein (Hsp-UTR). P-enoA142 enhanced the transcriptional level of a heterologous lipase gene and Hsp-UTR improved its translational efficiency. Fusarium heterosporum lipase (FHL) was inserted into a pSENSU-FHL expression vector harboring P-enoA142 and Hsp-UTR and was transformed into an A. oryzae NS4 strain. Transformants possessing pSENSU-FHL in single (pSENSU-FHL1) and double copies (pSENSU-FHL2) were selected to evaluate the lipase activity of the whole-cell biocatalyst. The two strains, pSENSU-FHL1 and 2, showed excellent lipase activity in hydrolysis compared with the strain transformed with conventional expression vector pNAN8142-FHL. Furthermore, by using pSENSU-FHL2, methanolysis could proceed much more effectively without deactivation, which allowed a swift addition of methanol to the reaction mixture, thereby reducing reaction time. (orig.)

  16. Biodegradation of methyl parathion by whole cells of marine-derived fungi Aspergillus sydowii and Penicillium decaturense.

    Science.gov (United States)

    Alvarenga, Natália; Birolli, Willian G; Seleghim, Mirna H R; Porto, André L M

    2014-12-01

    Seven marine fungi strains (Aspergillus sydowii CBMAI 934, A. sydowii CBMAI 935, A. sydowii CBMAI 1241, Penicillium decaturense CBMAI 1234, Penicillium raistrickii CBMAI 931, P. raistrickii CBMAI 1235, and Trichoderma sp. CBMAI 932) were screened by their growth in the presence of methyl parathion (MP) in a solid culture medium. The strains with best growth were A. sydowii CBMAI 935 and P. decaturense CBMAI 1234. Biodegradation reactions were performed in 10, 20 and 30d in a malt extract liquid medium containing commercial MP and whole cells of A. sydowii CBMAI 935 and P. decaturense CBMAI 1234. In 20d, A. sydowii CBMAI 935 was able to degrade all pesticide, whereas P. decaturense CBMAI 1234 promoted a complete degradation in 30d. A. sydowii CBMAI 935 and P. decaturense CBMAI 1234 could degrade the product of the MP enzymatic hydrolysis, p-nitrophenol, on average of 51 and 40% respectively. Both strains used MP as a sole source of carbon and provided satisfactory results. Metabolites detected in the medium showed that the presumable reaction pathway occurred through the activation of MP to its more toxic form, methyl paraoxon, which was further degraded to p-nitrophenol.

  17. Application of cell factory in prepation of varicella vaccine%细胞工厂在水痘疫苗生产中的应用

    Institute of Scientific and Technical Information of China (English)

    王亮; 杨月莲; 周锋; 谢蕾; 孙世杰; 彭晓; 周翎乔; 马相虎

    2012-01-01

    目的:提高水痘疫苗产量和厂房的利用率.方法:采用细胞工厂培养MRC-5细胞,以Roux瓶培养作对照,比较两种工艺的细胞生产情况、原液收获量等,并进行各项检定.结果:在同样细胞复苏量的条件下,采用细胞工厂比Roux瓶工艺生产的水痘疫苗产量要高,且各项检测指标均合格.结论:利用细胞工厂制备水痘疫苗,不仅提高了疫苗产量,而且节省了培养空间,降低了污染率,可用于水痘疫苗的大规模生产.%Objective:To improve the yield and plant utilization of varicella vaccine. Methods; MRC-5 cells were cultured on cell factory, using those cultured in Roux bottle as control. The growths of cells and yield of harvested virus cultured by the two methods were compared. The harvested virus liquid was concentrated by ultrafihration, then subjected to control tests. Results; In the condition of same quantity of recovery, varicella vaccine prepared by cell factory showed higher yield, compared with vaccine prepared by Roux bottle. The bulk was qualified in control tests. Conclusion:The yield of vaccine prepared by culture of MRC-5 cell on cell factory increased, while the cultural space and contamination decreased, indicating that cell factory might be used for large-scale production of varicella vaccine.

  18. A molecular analysis of L-arabinan degradation in Aspergillus niger and Aspergillus nidulans.

    NARCIS (Netherlands)

    Flipphi, M.J.A.

    1995-01-01

    This thesis describes a molecular study of the genetics ofL-arabinan degradation in Aspergillus niger and Aspergillus nidulans. These saprophytic hyphal fungi produce an extracellular hydrolytic enzyme system to depolymerize the plant cell wall polysaccharideL<

  19. Engineering propionibacteria as versatile cell factories for the production of industrially important chemicals: advances, challenges, and prospects.

    Science.gov (United States)

    Guan, Ningzi; Zhuge, Xin; Li, Jianghua; Shin, Hyun-Dong; Wu, Jing; Shi, Zhongping; Liu, Long

    2015-01-01

    Propionibacteria are actinobacteria consisting of two principal groups: cutaneous and dairy. Cutaneous propionibacteria are considered primary pathogens to humans, whereas dairy propionibacteria are widely used in the food and pharmaceutical industries. Increasing attention has been focused on improving the performance of dairy propionibacteria for the production of industrially important chemicals, and significant advances have been made through strain engineering and process optimization in the production of flavor compounds, nutraceuticals, and antimicrobial compounds. In addition, genome sequencing of several propionibacteria species has been completed, deepening understanding of the metabolic and physiological features of these organisms. However, the metabolic engineering of propionibacteria still faces several challenges owing to the lack of efficient genome manipulation tools and the existence of various types of strong restriction-modification systems. The emergence of systems and synthetic biology provides new opportunities to overcome these bottlenecks. In this review, we first introduce the major species of propionibacteria and their properties and provide an overview of their functions and applications. We then discuss advances in the genome sequencing and metabolic engineering of these bacteria. Finally, we discuss systems and synthetic biology approaches for engineering propionibacteria as efficient and robust cell factories for the production of industrially important chemicals.

  20. The Transcriptional Repressor TupA in Aspergillus niger Is Involved in Controlling Gene Expression Related to Cell Wall Biosynthesis, Development, and Nitrogen Source Availability

    DEFF Research Database (Denmark)

    Schachtschabel, Doreen; Arentshorst, Mark; Nitsche, Benjamin M

    2013-01-01

    The Tup1-Cyc8 (Ssn6) complex is a well characterized and conserved general transcriptional repressor complex in eukaryotic cells. Here, we report the identification of the Tup1 (TupA) homolog in the filamentous fungus Aspergillus niger in a genetic screen for mutants with a constitutive expression...... of the agsA gene. The agsA gene encodes a putative alpha-glucan synthase, which is induced in response to cell wall stress in A. niger. Apart from the constitutive expression of agsA, the selected mutant was also found to produce an unknown pigment at high temperatures. Complementation analysis...

  1. The Penicillium chrysogenum extracellular proteome. Conversion from a food-rotting strain to a versatile cell factory for white biotechnology.

    Science.gov (United States)

    Jami, Mohammad-Saeid; García-Estrada, Carlos; Barreiro, Carlos; Cuadrado, Abel-Alberto; Salehi-Najafabadi, Zahra; Martín, Juan-Francisco

    2010-12-01

    The filamentous fungus Penicillium chrysogenum is well-known by its ability to synthesize β-lactam antibiotics as well as other secondary metabolites. Like other filamentous fungi, this microorganism is an excellent host for secretion of extracellular proteins because of the high capacity of its protein secretion machinery. In this work, we have characterized the extracellular proteome reference map of P. chrysogenum Wisconsin 54-1255 by two-dimensional gel electrophoresis. This method allowed the correct identification of 279 spots by peptide mass fingerprinting and tandem MS. These 279 spots included 328 correctly identified proteins, which corresponded to 131 different proteins and their isoforms. One hundred and two proteins out of 131 were predicted to contain either classical or nonclassical secretion signal peptide sequences, providing evidence of the authentic extracellular location of these proteins. Proteins with higher representation in the extracellular proteome were those involved in plant cell wall degradation (polygalacturonase, pectate lyase, and glucan 1,3-β-glucosidase), utilization of nutrients (extracellular acid phosphatases and 6-hydroxy-d-nicotine oxidase), and stress response (catalase R). This filamentous fungus also secretes enzymes specially relevant for food industry, such as sulfydryl oxidase, dihydroxy-acid dehydratase, or glucoamylase. The identification of several antigens in the extracellular proteome also highlights the importance of this microorganism as one of the main indoor allergens. Comparison of the extracellular proteome among three strains of P. chrysogenum, the wild-type NRRL 1951, the Wis 54-1255 (an improved, moderate penicillin producer), and the AS-P-78 (a penicillin high-producer), provided important insights to consider improved strains of this filamentous fungus as versatile cell-factories of interest, beyond antibiotic production, for other aspects of white biotechnology.

  2. Development of microbial cell factories for bio-refinery through synthetic bioengineering.

    Science.gov (United States)

    Kondo, Akihiko; Ishii, Jun; Hara, Kiyotaka Y; Hasunuma, Tomohisa; Matsuda, Fumio

    2013-01-20

    Synthetic bioengineering is a strategy for developing useful microbial strains with innovative biological functions. Novel functions are designed and synthesized in host microbes with the aid of advanced technologies for computer simulations of cellular processes and the system-wide manipulation of host genomes. Here, we review the current status and future prospects of synthetic bioengineering in the yeast Saccharomyces cerevisiae for bio-refinery processes to produce various commodity chemicals from lignocellulosic biomass. Previous studies to improve assimilation of xylose and production of glutathione and butanol suggest a fixed pattern of problems that need to be solved, and as a crucial step, we now need to identify promising targets for further engineering of yeast metabolism. Metabolic simulation, transcriptomics, and metabolomics are useful emerging technologies for achieving this goal, making it possible to optimize metabolic pathways. Furthermore, novel genes responsible for target production can be found by analyzing large-scale data. Fine-tuning of enzyme activities is essential in the latter stage of strain development, but it requires detailed modeling of yeast metabolic functions. Recombinant technologies and genetic engineering are crucial for implementing metabolic designs into microbes. In addition to conventional gene manipulation techniques, advanced methods, such as multicistronic expression systems, marker-recycle gene deletion, protein engineering, cell surface display, genome editing, and synthesis of very long DNA fragments, will facilitate advances in synthetic bioengineering.

  3. Accurate Dna Assembly And Direct Genome Integration With Optimized Uracil Excision Cloning To Facilitate Engineering Of Escherichia Coli As A Cell Factory

    DEFF Research Database (Denmark)

    Cavaleiro, Mafalda; Kim, Se Hyeuk; Nørholm, Morten

    2015-01-01

    Plants produce a vast diversity of valuable compounds with medical properties, but these are often difficult to purify from the natural source or produce by organic synthesis. An alternative is to transfer the biosynthetic pathways to an efficient production host like the bacterium Escherichia co......-excision-based cloning and combining it with a genome-engineering approach to allow direct integration of whole metabolic pathways into the genome of E. coli, to facilitate the advanced engineering of cell factories....

  4. Optimization of date syrup for enhancement of the production of citric acid using immobilized cells of Aspergillus niger.

    Science.gov (United States)

    Mostafa, Yasser S; Alamri, Saad A

    2012-04-01

    Date syrup as an economical source of carbohydrates and immobilized Aspergillus niger J4, which was entrapped in calcium alginate pellets, were employed for enhancing the production of citric acid. Maximum production was achieved by pre-treating date syrup with 1.5% tricalcium phosphate to remove heavy metals. The production of citric acid using a pretreated medium was 38.87% higher than an untreated one that consumed sugar. The appropriate presence of nitrogen, phosphate and magnesium appeared to be important in order for citric acid to accumulate. The production of citric acid and the consumed sugar was higher when using 0.1% ammonium nitrate as the best source of nitrogen. The production of citric acid increased significantly when 0.1 g/l of KH2PO4 was added to the medium of date syrup. The addition of magnesium sulfate at the rate of 0.20 g/l had a stimulating effect on the production of citric acid. Maximum production of citric acid was obtained when calcium chloride was absent. One of the most important benefits of immobilized cells is their ability and stability to produce citric acid under a repeated batch culture. Over four repeated batches, the production of citric acid production was maintained for 24 days when each cycle continued for 144 h. The results obtained in the repeated batch cultivation using date syrup confirmed that date syrup could be used as a medium for the industrial production of citric acid.

  5. Integrated analysis of the global transcriptional response to α-amylase over-production by Aspergillus oryzae.

    Science.gov (United States)

    Vongsangnak, Wanwipa; Hansen, Kim; Nielsen, Jens

    2011-05-01

    The filamentous fungus Aspergillus oryzae is an important microbial cell factory for industrial production of many enzymes, such as α-amylase. In order to optimize the industrial enzyme production process, there is a need to understand fundamental processes underlying enzyme production, here under how enzyme production links to metabolism through global regulation. Through a genome-scale metabolic network for integrated analysis of transcriptome data and flux calculation, we identified key players (genes, enzymes, proteins, and metabolites) involved in the processes of enzyme synthesis and secretion, nucleotide metabolism, and amino acid metabolism that can be the potential targets for improving industrial enzyme production.

  6. A robust whole-cell biocatalyst that introduces a thermo- and solvent-tolerant lipase into Aspergillus oryzae cells: characterization and application to enzymatic biodiesel production.

    Science.gov (United States)

    Adachi, Daisuke; Koh, FookHee; Hama, Shinji; Ogino, Chiaki; Kondo, Akihiko

    2013-05-10

    To develop a robust whole-cell biocatalyst that works well at moderately high temperature (40-50°C) with organic solvents, a thermostable lipase from Geobacillus thermocatenulatus (BTL2) was introduced into an Aspergillus oryzae whole-cell biocatalyst. The lipase-hydrolytic activity of the immobilized A. oryzae (r-BTL) was highest at 50°C and was maintained even after an incubation of 24-h at 60°C. In addition, r-BTL was highly tolerant to 30% (v/v) organic solvents (dimethyl carbonate, ethanol, methanol, 2-propanol or acetone). The attractive characteristics of r-BTL also worked efficiently on palm oil methanolysis, resulting in a nearly 100% conversion at elevated temperature from 40 to 50°C. Moreover, r-BTL catalyzed methanolysis at a high methanol concentration without a significant loss of lipase activity. In particular, when 2 molar equivalents of methanol were added 2 times, a methyl ester content of more than 90% was achieved; the yield was higher than those of conventional whole-cell biocatalyst and commercial Candida antarctica lipase (Novozym 435). On the basis of the results regarding the excellent lipase characteristics and efficient biodiesel production, the developed whole-cell biocatalyst would be a promising biocatalyst in a broad range of applications including biodiesel production.

  7. Cell wall protein and glycoprotein constituents of Aspergillus fumigatus that bind to polystyrene may be responsible for the cell surface hydrophobicity of the mycelium.

    Science.gov (United States)

    Peñalver, M C; Casanova, M; Martínez, J P; Gil, M L

    1996-07-01

    Cell surface hydrophobicity (CSH) of Aspergillus fumigatus grown both in complex medium (yeast extract/peptone/dextrose; YPD) and minimal (Vogel's N) medium was monitored by assessing attachment of polystyrene microspheres to the cell surface. It was found that mature mycelium was hydrophobic. Treatment of intact mycelium with beta-mercaptoethanol (beta ME) abolished binding of the microspheres to hyphal elements, and coating of the microspheres with beta ME extracts from mycelium inhibited their attachment to intact mycelial cells. A. fumigatus mycelium was tagged in vivo with biotin and treated with beta ME. The beta ME extracts were analysed by SDS-PAGE and Western blotting with both peroxidase-conjugated-ExtrAvidin and concanavalin A (ConA). This procedure allowed identification of cell wall surface proteins and glycoproteins. Rabbit polyclonal antisera were raised against beta ME extracts obtained from cells grown in YPD and Vogel's N media. These antisera defined some major cell-wall-bound antigens. SDS-PAGE and Western blotting analysis of the cell wall material released by beta ME and adsorbed on polystyrene microspheres revealed about 19 protein species with apparent molecular masses ranging from 20 to 70 kDa, and two high-molecular-mass glycoproteins of 115 and 210 kDa. Treatment of cells grown in YPD, but not those grown in Vogel's N medium, with beta ME released a 55 kDa polypeptide able to adsorb to polystyrene microspheres that was detectable with the antisera. The ability to bind to polystyrene particles exhibited by several protein and glycoprotein species released by beta ME treatment suggested that these cell wall moieties possess exposed hydrophobic domains that could be responsible for the CSH of mycelium.

  8. Cytokine responses and regulation of interferon-gamma release by human mononuclear cells to Aspergillus fumigatus and other filamentous fungi.

    NARCIS (Netherlands)

    Warris, A.; Netea, M.G.; Verweij, P.E.; Gaustad, P.; Kullberg, B.J.; Weemaes, C.M.R.; Abrahamsen, T.G.

    2005-01-01

    There is substantial evidence that the production of proinflammatory cytokines is important in host resistance to invasive aspergillosis. Knowledge of the host response towards other filamentous fungi is scarce, as most studies have focused on Aspergillus fumigatus. In addition, interferon-gamma (IF

  9. Aspergillus thyroiditis: first antemortem case diagnosed by fine-needle aspiration culture in a pediatric stem cell transplant patient.

    Science.gov (United States)

    Badawy, S M; Becktell, K D; Muller, W J; Schneiderman, J

    2015-12-01

    Aspergillus thyroiditis (AT) has historically been considered a postmortem diagnosis in immunocompromised patients; most have disseminated disease. This report summarizes the clinical challenge of diagnosing AT. It also highlights the value of the early use of thyroid fine-needle aspiration culture and the need for a high index of suspicion to reach the final diagnosis before disease dissemination.

  10. Biocatalytic potential of laccase-like multicopper oxidases from Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Tamayo-Ramos Juan Antonio

    2012-12-01

    Full Text Available Abstract Background Laccase-like multicopper oxidases have been reported in several Aspergillus species but they remain uncharacterized. The biocatalytic potential of the Aspergillus niger fungal pigment multicopper oxidases McoA and McoB and ascomycete laccase McoG was investigated. Results The laccase-like multicopper oxidases McoA, McoB and McoG from the commonly used cell factory Aspergillus niger were homologously expressed, purified and analyzed for their biocatalytic potential. All three recombinant enzymes were monomers with apparent molecular masses ranging from 80 to 110 kDa. McoA and McoG resulted to be blue, whereas McoB was yellow. The newly obtained oxidases displayed strongly different activities towards aromatic compounds and synthetic dyes. McoB exhibited high catalytic efficiency with N,N-dimethyl-p-phenylenediamine (DMPPDA and 2,2-azino-di(3-ethylbenzthiazoline sulfonic acid (ABTS, and appeared to be a promising biocatalyst. Besides oxidizing a variety of phenolic compounds, McoB catalyzed successfully the decolorization and detoxification of the widely used textile dye malachite green. Conclusions The A. niger McoA, McoB, and McoG enzymes showed clearly different catalytic properties. Yellow McoB showed broad substrate specificity, catalyzing the oxidation of several phenolic compounds commonly present in different industrial effluents. It also harbored high decolorization and detoxification activity with the synthetic dye malachite green, showing to have an interesting potential as a new industrial biocatalyst.

  11. Aspergillus spinal epidural abscess

    Energy Technology Data Exchange (ETDEWEB)

    Byrd, B.F. III (Vanderbilt Univ. School of Medicine, Nashville, TN); Weiner, M.H.; McGee, Z.A.

    1982-12-17

    A spinal epidural abscess developed in a renal transplant recipient; results of a serum radioimmunoassay for Aspergillus antigen were positive. Laminectomy disclosed an abscess of the L4-5 interspace and L-5 vertebral body that contained hyphal forms and from which Aspergillus species was cultured. Serum Aspergillus antigen radioimmunoassay may be a valuable, specific early diagnostic test when systemic aspergillosis is a consideration in an immunosuppressed host.

  12. Increasing cell biomass in Saccharomyces cerevisiae increases recombinant protein yield: the use of a respiratory strain as a microbial cell factory

    Directory of Open Access Journals (Sweden)

    Hedfalk Kristina

    2010-06-01

    Full Text Available Abstract Background Recombinant protein production is universally employed as a solution to obtain the milligram to gram quantities of a given protein required for applications as diverse as structural genomics and biopharmaceutical manufacture. Yeast is a well-established recombinant host cell for these purposes. In this study we wanted to investigate whether our respiratory Saccharomyces cerevisiae strain, TM6*, could be used to enhance the productivity of recombinant proteins over that obtained from corresponding wild type, respiro-fermentative strains when cultured under the same laboratory conditions. Results Here we demonstrate at least a doubling in productivity over wild-type strains for three recombinant membrane proteins and one recombinant soluble protein produced in TM6* cells. In all cases, this was attributed to the improved biomass properties of the strain. The yield profile across the growth curve was also more stable than in a wild-type strain, and was not further improved by lowering culture temperatures. This has the added benefit that improved yields can be attained rapidly at the yeast's optimal growth conditions. Importantly, improved productivity could not be reproduced in wild-type strains by culturing them under glucose fed-batch conditions: despite having achieved very similar biomass yields to those achieved by TM6* cultures, the total volumetric yields were not concomitantly increased. Furthermore, the productivity of TM6* was unaffected by growing cultures in the presence of ethanol. These findings support the unique properties of TM6* as a microbial cell factory. Conclusions The accumulation of biomass in yeast cell factories is not necessarily correlated with a proportional increase in the functional yield of the recombinant protein being produced. The respiratory S. cerevisiae strain reported here is therefore a useful addition to the matrix of production hosts currently available as its improved biomass

  13. Fractional factorial plans

    CERN Document Server

    Dey, Aloke

    2009-01-01

    A one-stop reference to fractional factorials and related orthogonal arrays.Presenting one of the most dynamic areas of statistical research, this book offers a systematic, rigorous, and up-to-date treatment of fractional factorial designs and related combinatorial mathematics. Leading statisticians Aloke Dey and Rahul Mukerjee consolidate vast amounts of material from the professional literature--expertly weaving fractional replication, orthogonal arrays, and optimality aspects. They develop the basic theory of fractional factorials using the calculus of factorial arrangements, thereby providing a unified approach to the study of fractional factorial plans. An indispensable guide for statisticians in research and industry as well as for graduate students, Fractional Factorial Plans features: * Construction procedures of symmetric and asymmetric orthogonal arrays. * Many up-to-date research results on nonexistence. * A chapter on optimal fractional factorials not based on orthogonal arrays. * Trend-free plans...

  14. Epstein-Barr virus-transformed B-cells as efficient antigen presenting cells to propagate Aspergillus-specific cytotoxic T-lymphocytes.

    Science.gov (United States)

    Ramadan, Gamal

    2008-01-01

    To overcome the cytotoxic T-lymphocytes (CTL) expansion limitations imposed by the lack of sufficient dendritic cells (DC) alternative sources of autologous antigen presenting cells (APC) such as Epstein-Barr virus (EBV)-transformed B-lymphoblastoid cell lines (BLCL), which are easy to establish in vitro, have been considered and studied in the present work. Non-adherent peripheral blood mononuclear cells of three healthy donors were repeatedly primed with autologous Aspergillus fumigatus commercial culture-filtrate antigen-pulsed fast monocyte-derived DC (Aspf-CFA-DC) alone, Aspf-CFA-pulsed BLCL (Aspf-CFA-BLCL) alone or Aspf-CFA-BLCL after one, two, or three primings with Aspf-CFA-DC (1DC/BLCL, 2DC/BLCL or 3DCIBLCL; respectively). After 5th priming, lines generated by Aspf-CFA-BLCL only showed strong/weak lytic activity for EBV/Aspf; respectively. Aspf-specific lytic activity in all donors was increased by increasing the number of primings with Aspf-CFA-DC before switching to Aspf-CFA-BLCL (18.20 +/- 1.65% versus 35.67 +/- 1.02% and 40.03 +/- 1.41% in bulk cultures generated by 1DC/BLCL versus 2DC/BLCL and 3DC/BLCL, respectively). Bulk cultures generated by Aspf-CFA-BLCL after at least two primings with Aspf-CFA-DC showed approximately the same Aspf-specific lytic activity, effector cell phenotype, expansion level and percentage expression of IFN-gamma, CD69 and CD107a without any significant differences (p > 0.05) as standard bulk cultures generated by only Aspf-CFA-DC. Thus, this study explored the use of a combined DC/BLCL protocol to establish/propagate Aspf-specific CTL for adoptive immunotherapy to prevent or treat invasive pulmonary aspergillosis.

  15. Optimization of date syrup for enhancement of the production of citric acid using immobilized cells of Aspergillus niger

    OpenAIRE

    Mostafa, Yasser S.; Alamri, Saad A.

    2012-01-01

    Date syrup as an economical source of carbohydrates and immobilized Aspergillus niger J4, which was entrapped in calcium alginate pellets, were employed for enhancing the production of citric acid. Maximum production was achieved by pre-treating date syrup with 1.5% tricalcium phosphate to remove heavy metals. The production of citric acid using a pretreated medium was 38.87% higher than an untreated one that consumed sugar. The appropriate presence of nitrogen, phosphate and magnesium appear...

  16. The stimulatory effect of growth hormone, prolactin, and placental lactogen on beta-cell proliferation is not mediated by insulin-like growth factor-I

    DEFF Research Database (Denmark)

    Billestrup, N; Nielsen, Jens Høiriis

    1991-01-01

    The effects of GH, PRL, and placental lactogen (PL) on the proliferation of pancreatic beta-cells in vitro were studied as well as the possible effect of insulin-like growth factor-I (IGF-I) in mediating this effect. Proliferating beta-cells were identified by staining with a monoclonal antibody...... to that caused by hGH. GH, PRL, and PL had no effect on the proliferation of glucagon- or somatostatin-producing cells. The addition of 100 ng/ml IGF-I to either control or GH-stimulated islet cells did not affect the labeling index. When GH-stimulated proliferation of beta-cells was measured in the presence...

  17. Stimulation by means of dendritic cells followed by Epstein-Barr virus-transformed B cells as antigen-presenting cells is more efficient than dendritic cells alone in inducing Aspergillus f16-specific cytotoxic T cell responses.

    Science.gov (United States)

    Zhu, F; Ramadan, G; Davies, B; Margolis, D A; Keever-Taylor, C A

    2008-02-01

    Adoptive immunotherapy with in vitro expanded antigen-specific cytotoxic T lymphocytes (CTLs) may be an effective approach to prevent, or even treat, Aspergillus (Asp) infections. Such lines can be generated using monocyte-derived dendritic cells (DC) as antigen-presenting cells (APC) but requires a relatively high volume of starting blood. Here we describe a method that generates Asp-specific CTL responses more efficiently using a protocol of antigen presented on DC followed by Epstein-Barr virus (EBV)-transformed B lymphoblastoid cell lines (BLCL) as APC. Peripheral blood mononuclear cells were stimulated weekly (2-5x) with a complete pool of pentadecapeptides (PPC) spanning the coding region of Asp f16 pulsed onto autologous mature DC. Cultures were split and stimulated subsequently with either PPC-DC or autologous PPC-pulsed BLCL (PPC-BLCL). Lines from the DC/BLCL arm demonstrated Asp f16-specific cytotoxicity earlier and to a higher degree than lines generated with PPC-DC alone. The DC/BLCL-primed lines showed a higher frequency of Asp f16-specific interferon (IFN)-gamma producing cells but an identical effector cell phenotype and peptide specificity compared to PPC-DC-only-primed lines. Tumour necrosis factor (TNF)-alpha, but not IL-10, appeared to play a role in the effectiveness of BLCL as APC. These results demonstrate that BLCL serve as highly effective APC for the stimulation of Asp f16-specific T cell responses and that a culture approach using initial priming with PPC-DC followed by PPC-BLCL may be a more effective method to generate Asp f16-specific T cell lines and requires less starting blood than priming with PPC-DC alone.

  18. Metabolomics of Aspergillus fumigatus

    DEFF Research Database (Denmark)

    Frisvad, Jens Christian; Rank, Christian; Nielsen, Kristian Fog

    2009-01-01

    Aspergillus fumigatus is the most important species in Aspergillus causing infective lung diseases. This species has been reported to produce a large number of extrolites, including secondary metabolites, acids, and proteins such as hydrophobins and extracellular enzymes. At least 226 potentially...

  19. Natural killer cell activity, lymphocyte proliferation, and cytokine profile in tumor-bearing mice treated with MAPA, a magnesium aggregated polymer from Aspergillus oryzae.

    Science.gov (United States)

    Justo, G Z; Durán, N; Queiroz, M L S

    2003-08-01

    The present study examined the effects of MAPA, an antitumor aggregated polymer of protein magnesium ammonium phospholinoleate-palmitoleate anhydride, isolated from Aspergillus oryzae, on concanavalin A (Con A)-induced spleen cell proliferation, cytokine production and on natural killer (NK) cell activity in Ehrlich ascites tumor-bearing mice. The Ehrlich ascites tumor (EAT) growth led to diminished mitogen-induced expansion of spleen cell populations and total NK activity. This was accompanied by striking spleen enlargement, with a marked increase in total cell counts. Moreover, a substantial enhancement in IL-10 levels, paralleled by a significant decrease in IL-2 was observed, while production of IL-4 and interferon-gamma (IFN-gamma) was not altered. Treatment of mice with 5 mg/kg MAPA for 7 days promoted spleen cell proliferation, IL-2 production and NK cell activity regardless of tumor outgrowth. In addition, MAPA treatment markedly enhanced IFN-gamma levels and reduced IL-10 production relative to EAT mice. A 35% reduction in splenomegaly with normal number of nucleated cells was also found. Altogether, our results suggest that MAPA directly and/or indirectly modulates immune cell activity, and probably disengages tumor-induced suppression of these responses. Clearly, MAPA has an impact and may delay tumor outgrowth through immunotherapeutic mechanisms.

  20. Characterization of pectate lyase A from Aspergillus niger

    NARCIS (Netherlands)

    Benen, J.A.E.; Parenicova, L.; Kester, H.C.M.; Visser, J.

    2001-01-01

    The Aspergillus niger plyA gene encoding pectate lyase A (EC 4.2.99.3) was cloned from a chromosomal EMBL4 library using the Aspergillus nidulans pectate lyase encoding gene [Dean, R. A., and Timberlake, W. E. (1989) Plant Cell 1, 275-284] as a probe. The plyA gene was overexpressed using a promoter

  1. Fermented Brown Rice and Rice Bran with Aspergillus oryzae (FBRA Prevents Inflammation-Related Carcinogenesis in Mice, through Inhibition of Inflammatory Cell Infiltration

    Directory of Open Access Journals (Sweden)

    Kunishige Onuma

    2015-12-01

    Full Text Available We have established an inflammation-related carcinogenesis model in mouse, in which regressive QR-32 cells subcutaneously co-implanted with a foreign body—gelatin sponge—convert themselves into lethal tumors due to massive infiltration of inflammatory cells into the sponge. Animals were fed with a diet containing 5% or 10% fermented brown rice and rice bran with Aspergillus oryzae (FBRA. In 5% and 10% FBRA diet groups, tumor incidences were lower (35% and 20%, respectively than in the non-treated group (70%. We found that FBRA reduced the number of inflammatory cells infiltrating into the sponge. FBRA administration did not cause myelosuppression, which indicated that the anti-inflammatory effects of FBRA took place at the inflammatory lesion. FBRA did not have antitumor effects on the implanted QRsP-11 tumor cells, which is a tumorigenic cell line established from a tumor arisen after co-implantation of QR-32 cells with sponge. FBRA did not reduce formation of 8-hydroxy-2′-deoxyguanine adducts, a marker of oxidative DNA damage in the inflammatory lesion; however, it reduced expression of inflammation-related genes such as TNF-α, Mac-1, CCL3 and CXCL2. These results suggest that FBRA will be an effective chemopreventive agent against inflammation-related carcinogenesis that acts by inhibiting inflammatory cell infiltration into inflammatory lesions.

  2. PCs in the factory

    CERN Document Server

    2013-01-01

    Please note this is a short discount publication.PCs have become as essential to the factory environment as they are to the office environment. This in-depth report examines how specially adapted PCs and peripherals are being established in Factory Process Control and Reporting. The report covers: * Hardware and Software* Typical Applications* Implementation Issues* Case Studies and Real Applications

  3. Railways Factory in Resita

    Directory of Open Access Journals (Sweden)

    Cornel Iacob-Mare

    2013-09-01

    Full Text Available This paper proposes an exemption Analysis railway factory in Resita impressed having experience and also provide some technical characteristics of freight wagons representative technology, used to transport products made in factories in ReşiŃa.

  4. A WDR Gene Is a Conserved Member of a Chitin Synthase Gene Cluster and Influences the Cell Wall in Aspergillus nidulans

    Directory of Open Access Journals (Sweden)

    Gea Guerriero

    2016-06-01

    Full Text Available WD40 repeat (WDR proteins are pleiotropic molecular hubs. We identify a WDR gene that is a conserved genomic neighbor of a chitin synthase gene in Ascomycetes. The WDR gene is unique to fungi and plants, and was called Fungal Plant WD (FPWD. FPWD is within a cell wall metabolism gene cluster in the Ascomycetes (Pezizomycotina comprising chsD, a Chs activator and a GH17 glucanase. The FPWD, AN1556.2 locus was deleted in Aspergillus nidulans strain SAA.111 by gene replacement and only heterokaryon transformants were obtained. The re-annotation of Aspergilli genomes shows that AN1556.2 consists of two tightly linked separate genes, i.e., the WDR gene and a putative beta-flanking gene of unknown function. The WDR and the beta-flanking genes are conserved genomic neighbors localized within a recently identified metabolic cell wall gene cluster in genomes of Aspergilli. The heterokaryons displayed increased susceptibility to drugs affecting the cell wall, and their phenotypes, observed by optical, confocal, scanning electron and atomic force microscopy, suggest cell wall alterations. Quantitative real-time PCR shows altered expression of some cell wall-related genes. The possible implications on cell wall biosynthesis are discussed.

  5. Genomic islands in the pathogenic filamentous fungus Aspergillus fumigatus.

    Directory of Open Access Journals (Sweden)

    Natalie D Fedorova

    2008-04-01

    Full Text Available We present the genome sequences of a new clinical isolate of the important human pathogen, Aspergillus fumigatus, A1163, and two closely related but rarely pathogenic species, Neosartorya fischeri NRRL181 and Aspergillus clavatus NRRL1. Comparative genomic analysis of A1163 with the recently sequenced A. fumigatus isolate Af293 has identified core, variable and up to 2% unique genes in each genome. While the core genes are 99.8% identical at the nucleotide level, identity for variable genes can be as low 40%. The most divergent loci appear to contain heterokaryon incompatibility (het genes associated with fungal programmed cell death such as developmental regulator rosA. Cross-species comparison has revealed that 8.5%, 13.5% and 12.6%, respectively, of A. fumigatus, N. fischeri and A. clavatus genes are species-specific. These genes are significantly smaller in size than core genes, contain fewer exons and exhibit a subtelomeric bias. Most of them cluster together in 13 chromosomal islands, which are enriched for pseudogenes, transposons and other repetitive elements. At least 20% of A. fumigatus-specific genes appear to be functional and involved in carbohydrate and chitin catabolism, transport, detoxification, secondary metabolism and other functions that may facilitate the adaptation to heterogeneous environments such as soil or a mammalian host. Contrary to what was suggested previously, their origin cannot be attributed to horizontal gene transfer (HGT, but instead is likely to involve duplication, diversification and differential gene loss (DDL. The role of duplication in the origin of lineage-specific genes is further underlined by the discovery of genomic islands that seem to function as designated "gene dumps" and, perhaps, simultaneously, as "gene factories".

  6. Photon collider Higgs factories

    CERN Document Server

    Telnov, V I

    2014-01-01

    The discovery of the Higgs boson (and still nothing else) have triggered appearance of many proposals of Higgs factories for precision measurement of the Higgs properties. Among them there are several projects of photon colliders (PC) without e+e- in addition to PLC based on e+e- linear colliders ILC and CLIC. In this paper, following a brief discussion of Higgs factories physics program I give an overview of photon colliders based on linear colliders ILC and CLIC, and of the recently proposed photon-collider Higgs factories with no e+e- collision option based on recirculation linacs in ring tunnels.

  7. Towards energy transparent factories

    CERN Document Server

    Posselt, Gerrit

    2016-01-01

    This monograph provides a methodological approach for establishing demand-oriented levels of energy transparency of factories. The author presents a systematic indication of energy drivers and cost factors, taking into account the interdependencies between facility and production domains. Particular attention is given to energy flow metering and monitoring. Readers will also be provided with an in-depth description of a planning tool which allows for systematically deriving suitable metering points in complex factory environments. The target audience primarily comprises researchers and experts in the field of factory planning, but the book may also be beneficial for graduate students.

  8. Deletion of glucose oxidase changes the pattern of organic acid production in Aspergillus carbonarius.

    Science.gov (United States)

    Yang, Lei; Lübeck, Mette; Lübeck, Peter S

    2014-01-01

    Aspergillus carbonarius has potential as a cell factory for the production of different organic acids. At pH 5.5, A.carbonarius accumulates high amounts of gluconic acid when it grows on glucose based medium whereas at low pH, it produces citric acid. The conversion of glucose to gluconic acid is carried out by secretion of the enzyme, glucose oxidase. In this work, the gene encoding glucose oxidase was identified and deleted from A. carbonarius with the aim of changing the carbon flux towards other organic acids. The effect of genetic engineering was examined by testing glucose oxidase deficient (Δgox) mutants for the production of different organic acids in a defined production medium. The results obtained showed that the gluconic acid accumulation was completely inhibited and increased amounts of citric acid, oxalic acid and malic acid were observed in the Δgox mutants.

  9. Gene deletion of cytosolic ATP: citrate lyase leads to altered organic acid production in Aspergillus niger

    DEFF Research Database (Denmark)

    Meijer, Susan Lisette; Nielsen, Michael Lynge; Olsson, Lisbeth

    2009-01-01

    With the availability of the genome sequence of the filamentous fungus Aspergillus niger, the use of targeted genetic modifications has become feasible. This, together with the fact that A. niger is well established industrially, makes this fungus an attractive micro-organism for creating a cell...... factory platform for production of chemicals. Using molecular biology techniques, this study focused on metabolic engineering of A. niger to manipulate its organic acid production in the direction of succinic acid. The gene target for complete gene deletion was cytosolic ATP: citrate lyase (acl), which...... the acl gene. Additionally, the total amount of organic acids produced in the deletion strain was significantly increased. Genome-scale stoichiometric metabolic model predictions can be used for identifying gene targets. Deletion of the acl led to increased succinic acid production by A. niger....

  10. Conidial Hydrophobins of Aspergillus fumigatus

    OpenAIRE

    Paris, Sophie; Debeaupuis, Jean-Paul; Crameri, Reto; Carey, Marilyn; Charlès, Franck; Prévost, Marie Christine; Schmitt, Christine; Philippe, Bruno; Latgé, Jean Paul

    2003-01-01

    The surface of Aspergillus fumigatus conidia, the first structure recognized by the host immune system, is covered by rodlets. We report that this outer cell wall layer contains two hydrophobins, RodAp and RodBp, which are found as highly insoluble complexes. The RODA gene was previously characterized, and ΔrodA conidia do not display a rodlet layer (N. Thau, M. Monod, B. Crestani, C. Rolland, G. Tronchin, J. P. Latgé, and S. Paris, Infect. Immun. 62:4380-4388, 1994). The RODB gene was cloned...

  11. Factorial PD-Clustering

    CERN Document Server

    Tortora, Cristina; Summa, Mireille Gettler

    2011-01-01

    Factorial clustering methods have been developed in recent years thanks to the improving of computational power. These methods perform a linear transformation of data and a clustering on transformed data optimizing a common criterion. Factorial PD-clustering is based on Probabilistic Distance clustering (PD-clustering). PD-clustering is an iterative, distribution free, probabilistic, clustering method. Factorial PD-clustering make a linear transformation of original variables into a reduced number of orthogonal ones using a common criterion with PD-Clustering. It is demonstrated that Tucker 3 decomposition allows to obtain this transformation. Factorial PD-clustering makes alternatively a Tucker 3 decomposition and a PD-clustering on transformed data until convergence. This method could significantly improve the algorithm performance and allows to work with large dataset, to improve the stability and the robustness of the method.

  12. Maximum Autocorrelation Factorial Kriging

    DEFF Research Database (Denmark)

    Nielsen, Allan Aasbjerg; Conradsen, Knut; Pedersen, John L.

    2000-01-01

    This paper describes maximum autocorrelation factor (MAF) analysis, maximum autocorrelation factorial kriging, and its application to irregularly sampled stream sediment geochemical data from South Greenland. Kriged MAF images are compared with kriged images of varimax rotated factors from...

  13. Virtual Factory Testbed

    Data.gov (United States)

    Federal Laboratory Consortium — The Virtual Factory Testbed (VFT) is comprised of three physical facilities linked by a standalone network (VFNet). The three facilities are the Smart and Wireless...

  14. Defective phagocyte Aspergillus killing associated with recurrent pulmonary Aspergillus infections.

    Science.gov (United States)

    Fietta, A; Sacchi, F; Mangiarotti, P; Manara, G; Gialdroni Grassi, G

    1984-01-01

    An apparently healthy boy was suffering from recurrent Aspergillus infections. No classical conditions of immunodeficiency were found. Studies on the patient's phagocytic system revealed neutrophils and monocytes to function normally except in Aspergillus killing (microbicidal activity for bacteria and Candida was normal). Aspergillus killing mechanisms may be complex and peculiarly selective, possibly involving both oxygen-dependent and independent mechanisms.

  15. Characterisation of Aspergillus niger prolyl aminopeptidase

    NARCIS (Netherlands)

    Basten, E.J.W.; Moers, A.P.H.A.; Ooyen, van A.J.J.; Schaap, P.J.

    2005-01-01

    We have cloned a gene (papA) that encodes a prolyl aminopeptidase from Aspergillus niger. Homologous genes are present in the genomes of the Eurotiales A. nidulans, A. fumigatus and Talaromyces emersonii, but the gene is not present in the genome of the yeast Saccharomyces cerevisiae. Cell extracts

  16. Dual silencing of insulin-like growth factor-I receptor and epidermal growth factor receptor in colorectal cancer cells is associated with decreased proliferation and enhanced apoptosis.

    Science.gov (United States)

    Kaulfuss, Silke; Burfeind, Peter; Gaedcke, Jochen; Scharf, Jens-Gerd

    2009-04-01

    Overexpression and activation of tyrosine kinase receptors are common features of colorectal cancer. Using the human colorectal cancer cell lines DLD-1 and Caco-2, we evaluated the role of the insulin-like growth factor-I (IGF-I) receptor (IGF-IR) and epidermal growth factor receptor (EGFR) in cellular functions of these cells. We used the small interfering RNA (siRNA) technology to specifically down-regulate IGF-IR and EGFR expression. Knockdown of IGF-IR and EGFR resulted in inhibition of cell proliferation of DLD-1 and Caco-2 cells. An increased rate of apoptosis was associated with siRNA-mediated silencing of IGF-IR and EGFR as assessed by activation of caspase-3/caspase-7. The combined knockdown of both EGFR and IGF-IR decreased cell proliferation and induced cell apoptosis more effectively than did silencing of either receptor alone. Comparable effects on cell proliferation and apoptosis were observed after single and combinational treatment of cells by the IGF-IR tyrosine kinase inhibitor NVP-AEW541 and/or the EGFR tyrosine kinase inhibitor erlotinib. Combined IGF-IR and EGFR silencing by either siRNAs or tyrosine kinase inhibitors diminished the phosphorylation of downstream signaling pathways AKT and extracellular signal-regulated kinase (ERK)-1/2 more effectively than did the single receptor knockdown. Single IGF-IR knockdown inhibited IGF-I-dependent phosphorylation of AKT but had no effect on IGF-I- or EGF-dependent phosphorylation of ERK1/2, indicating a role of EGFR in ligand-dependent ERK1/2 phosphorylation. The present data show that inhibition of the IGF-IR transduction cascade augments the antipoliferative and proapoptotic effects of EGFR inhibition in colorectal cancer cells. A clinical application of combination therapy targeting both EGFR and IGF-IR could be a promising therapeutic strategy.

  17. Aspergillus fumigatus germ tube growth and not conidia ingestion induces expression of inflammatory mediator genes in the human lung epithelial cell line A549.

    Science.gov (United States)

    Bellanger, Anne-Pauline; Millon, Laurence; Khoufache, Khaled; Rivollet, Danièle; Bièche, Ivan; Laurendeau, Ingrid; Vidaud, Michel; Botterel, Françoise; Bretagne, Stéphane

    2009-02-01

    Inhalation of conidia is the main cause of invasive pulmonary aspergillosis (IPA) and the respiratory epithelium is the first line of defence. To explore the triggering factor for the inflammatory response to Aspergillus fumigatus, the species mainly responsible for IPA, this study analysed the differential expression of three inflammatory genes in A549 cells after challenge with live and killed conidia. The influence of steroids, one of the main risk factors for developing IPA, was also investigated. Quantification of mRNAs of the inflammatory mediator genes encoding interleukin (IL)-8, tumour necrosis factor (TNF)-alpha and granulocyte-monocyte colony-stimulating factor (GM-CSF) was carried out using real-time PCR. Ingestion rates were studied for the conidia of A. fumigatus and Penicillium chrysogenum using a fluorescence brightener. Similar results were obtained for both species, with ingestion rates ranging from 35 to 40 %. Exposure of A549 cells to live A. fumigatus conidia only induced a four- to fivefold increase in the mRNA levels of the three genes, starting 8 h after the initial contact. Both inactivation of live A. fumigatus conidia and treatment by dexamethasone (10(-7) M) prevented the overexpression of TNF-alpha, IL-8 and GM-CSF. Fungal growth, rather than conidia ingestion, appears to be the main stimulus for the production of inflammatory mediators by epithelial cells, and this production is inhibited by steroid therapy. These results underline the role that the epithelium plays in the innate response against IPA.

  18. The evaluation of microfungal contamination of dust created during woodworking in furniture factories

    Directory of Open Access Journals (Sweden)

    Tomasz Rogoziński

    2014-12-01

    Full Text Available Background: Microscopic fungi are the biological agent of occupational risk in the woodworking environment. Microbiological and chemical methods were used for determination of their concentration and species composition in dust. Material and Methods: Dust was sampled in 3 factories producing furniture using different materials. The 1st factory (A processes solid wood, the 2nd (B – chipboards and the 3rd factory (C uses both wood and wood composites. The samples were collected in 12 different workstations and locations in each factory. The quantitative content of fungal biomass was determined basing on analysis of ergosterol (ERG. The species composition of fungi was analyzed using the microbiological method basing on culture morphology. Results: The concentration of ergosterol was relatively low and ranged from 0.012 mg/kg to 3.36 mg/kg. The average value of ERG amounted to 1.25 mg/kg in factories A and C and 1.15 mg/kg in factory B. The most frequently isolated fungi in factory A and B were Penicillum and Aspergillus. However, in the factory C, only Trichoderma was isolated. The maximum concentration of fungi in dust collected in factory B was 2377 cfu/g and it is 3 times more than in the dust from factories A and C. Conclusions: Workers of furniture factories may be exposed to airborne fungi associated with the wood dust posing health hazard. The content of these fungi is relatively small (ERG – max: 3.36 mg/kg but the species, especially genera Penicillum and Aspergillus, found in the dust which were reported as having allergic and toxic properties. Med Pr 2014;65(6:705–713

  19. Distinct galactofuranose antigens in the cell wall and culture supernatants as a means to differentiate Fusarium from Aspergillus species.

    Science.gov (United States)

    Wiedemann, Annegret; Kakoschke, Tamara Katharina; Speth, Cornelia; Rambach, Günter; Ensinger, Christian; Jensen, Henrik Elvang; Ebel, Frank

    2016-09-01

    Detection of carbohydrate antigens is an important means for diagnosis of invasive fungal infections. For diagnosis of systemic Aspergillus infections, galactomannan is commonly used, the core antigenic structure of which consists of chains of several galactofuranose moieties. In this study, we provide evidence that Fusarium produces at least two distinct galactofuranose antigens: Smaller amounts of galactomannan and larger quantities of a novel antigen recognized by the monoclonal antibody AB135-8. In A. fumigatus, only minor amounts of the AB135-8 antigen are found in supernatants and in the apical regions of hyphae. A galactofuranose-deficient A. fumigatus mutant lacks the AB135-8 antigen, which strongly suggests that galactofuranose is an essential constituent of this antigen. Using a combination of AB135-8 and a galactomannan-specific antibody, we were able to unambiguously differentiate A. fumigatus and Fusarium hyphae in immunohistology. Moreover, since Fusarium releases the AB135-8 antigen, it appears to be a promising target antigen for a serological detection of Fusarium infections.

  20. Azole-resistant Aspergillus fumigatus isolate with the TR34/L98H mutation in both a fungicide-sprayed field and the lung of a hematopoietic stem cell transplant recipient with invasive aspergillosis.

    Science.gov (United States)

    Rocchi, Steffi; Daguindau, Etienne; Grenouillet, Frédéric; Deconinck, Eric; Bellanger, Anne-Pauline; Garcia-Hermoso, Dea; Bretagne, Stéphane; Reboux, Gabriel; Millon, Laurence

    2014-05-01

    A French farmer developed invasive aspergillosis with azole-resistant Aspergillus fumigatus with the TR34/L98H mutation following a hematopoietic stem cell transplantation. He had worked in fungicide-sprayed fields where a non-genetically related A. fumigatus TR34/L98H isolate was collected. If azole resistance detection increases, voriconazole as first-line therapy might be questioned in agricultural areas.

  1. Mapping N-linked Glycosylation Sites in the Secretome and Whole Cells of Aspergillus niger Using Hydrazide Chemistry and Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Lu; Aryal, Uma K.; Dai, Ziyu; Mason, Alisa C.; Monroe, Matthew E.; Tian, Zhixin; Zhou, Jianying; Su, Dian; Weitz, Karl K.; Liu, Tao; Camp, David G.; Smith, Richard D.; Baker, Scott E.; Qian, Weijun

    2012-01-01

    Protein glycosylation is known to play an essential role in both cellular functions and the secretory pathways; however, little information is available on the dynamics of glycosylated N-linked glycosites of fungi. Herein we present the first extensive mapping of glycosylated N-linked glycosites in industrial strain Aspergillus niger by applying an optimized solid phase enrichment of glycopeptide protocol using hydrazide modified magnetic beads. The enrichment protocol was initially optimized using mouse plasma and A. niger secretome samples, which was then applied to profile N-linked glycosites from both the secretome and whole cell lysates of A. niger. A total of 847 unique N-linked glycosites and 330 N-linked glycoproteins were confidently identified by LC-MS/MS. Based on gene ontology analysis, the identified N-linked glycoproteins in the whole cell lysate were primarily localized in the plasma membrane, endoplasmic reticulum, golgi apparatus, lysosome, and storage vacuoles. The identified N-linked glycoproteins are involved in a wide range of biological processes including gene regulation and signal transduction, protein folding and assembly, protein modification and carbohydrate metabolism. The extensive coverage of glycosylated N-linked glycosites along with identification of partial N-linked glycosylation in those enzymes involving in different biochemical pathways provide useful information for functional studies of N-linked glycosylation and their biotechnological applications in A. niger.

  2. Production of cellulose- and xylan-degrading enzymes by a koji mold, aspergillus oryzae, and their contribution to the maceration of rice endosperm cell wall.

    Science.gov (United States)

    Yamane, Yu-Ichi; Fujita, Jin; Shimizu, Ryu-Ichi; Hiyoshi, Akira; Fukuda, Hisashi; Kizaki, Yasuzo; Wakabayashi, Saburo

    2002-01-01

    The production of cellulose- (CEL), xylan- (XYL), and pectin-degrading enzymes (PEC) by a koji mold, Aspergillus oryzae, was studied, and their contributions to the maceration of the rice endosperm cell wall were investigated with regard to the utilization of available rice in the sake mash. The sake koji mold showed higher CEL and XYL productivities, whereas the miso and soy sauce koji molds showed higher PEC productivity. Statistical analyses indicated that CEL and XYL contribute predominantly and synergistically to the maceration of the rice endosperm cell wall. A. oryzae produced at least three kinds of CEL (Cel-1, 2, 3) and two kinds of XYL (Xyl-1, 2) when cultured in a wheat bran medium. In the solid-state culture, the production of Cel-3 and Xyl-2 was markedly stimulated by decreasing the moisture content of the solid substrate, although the production levels of Cel-1 and Xyl-1 were almost the same. These data suggest that the production of Cel-3 and Xyl-2 is strongly influenced by culture conditions, and that water activity is one of the dominant factors in the regulation of their production.

  3. A pericentrin-related protein homolog in Aspergillus nidulans plays important roles in nucleus positioning and cell polarity by affecting microtubule organization.

    Science.gov (United States)

    Chen, Peiying; Gao, Rongsui; Chen, Shaochun; Pu, Li; Li, Pin; Huang, Ying; Lu, Ling

    2012-12-01

    Pericentrin is a large coiled-coil protein in mammalian centrosomes that serves as a multifunctional scaffold for anchoring numerous proteins. Recent studies have linked numerous human disorders with mutated or elevated levels of pericentrin, suggesting unrecognized contributions of pericentrin-related proteins to the development of these disorders. In this study, we characterized AnPcpA, a putative homolog of pericentrin-related protein in the model filamentous fungus Aspergillus nidulans, and found that it is essential for conidial germination and hyphal development. Compared to the hyphal apex localization pattern of calmodulin (CaM), which has been identified as an interactive partner of the pericentrin homolog, GFP-AnPcpA fluorescence dots are associated mainly with nuclei, while the accumulation of CaM at the hyphal apex depends on the function of AnPcpA. In addition, the depletion of AnPcpA by an inducible alcA promoter repression results in severe growth defects and abnormal nuclear segregation. Most interestingly, in mature hyphal cells, knockdown of pericentrin was able to significantly induce changes in cell shape and cytoskeletal remodeling; it resulted in some enlarged compartments with condensed nuclei and anucleate small compartments as well. Moreover, defects in AnPcpA significantly disrupted the microtubule organization and nucleation, suggesting that AnPcpA may affect nucleus positioning by influencing microtubule organization.

  4. Humoral and Cell-mediated Autoimmune Reactions to Human Acidic Ribosomal P2 Protein in Individuals Sensitized to Aspergillus fumigatus P2 Protein

    Science.gov (United States)

    Mayer, Christina; Appenzeller, Ulrich; Seelbach, Heike; Achatz, Gernot; Oberkofler, Hannes; Breitenbach, Michael; Blaser, Kurt; Crameri, Reto

    1999-01-01

    A panel of cDNAs encoding allergenic proteins was isolated from an Aspergillus fumigatus cDNA library displayed on the surface of filamentous phage. Solid phase–immobilized serum immunoglobulin E (IgE) from A. fumigatus–allergic individuals was used to enrich phage displaying IgE-binding molecules. One of the cDNAs encoded a 11.1-kD protein that was identified as acidic ribosomal phosphoprotein type 2 (P2 protein). The allergen, formally termed rAsp f 8, shares >62% sequence identity and >84% sequence homology to corresponding eukaryotic P2 proteins, including human P2 protein. The sequences encoding human and fungal P2 protein were subcloned, expressed in Escherichia coli as His6-tagged fusion proteins, and purified by Ni2+–chelate affinity chromatography. Both recombinant P2 proteins were recognized by IgE antibodies from allergic individuals sensitized to the A. fumigatus P2 protein and elicited strong type 1–specific skin reactions in these individuals. Moreover, human and fungal P2 proteins induced proliferative responses in peripheral blood mononuclear cells of A. fumigatus– allergic subjects sensitized to the fungal P2 protein. These data provide strong evidence for in vitro and in vivo humoral and cell-mediated autoreactivity to human P2 protein in patients suffering from chronic A. fumigatus allergy. PMID:10224291

  5. A novel unidirectional cross-talk from the insulin-like growth factor-I receptor to leptin receptor in human breast cancer cells.

    Science.gov (United States)

    Ozbay, Tuba; Nahta, Rita

    2008-06-01

    Obesity is a major risk factor for the development and progression of breast cancer. Increased circulating levels of the obesity-associated hormones leptin and insulin-like growth factor-I (IGF-I) and overexpression of the leptin receptor (Ob-R) and IGF-I receptor (IGF-IR) have been detected in a majority of breast cancer cases and during obesity. Due to correlations between increased leptin, Ob-R, IGF-I, and IGF-IR in breast cancer, we hypothesized that molecular interactions may exist between these two signaling pathways. Coimmunoprecipitation and immunoblotting showed that IGF-IR and Ob-R interact in the breast cancer cell lines MDA-MB-231, MCF7, BT474, and SKBR3. Stimulation of cells with IGF-I promoted Ob-R phosphorylation, which was blocked by IGF-IR kinase inhibition. In addition, IGF-I activated downstream signaling molecules in the leptin receptor and IGF-IR pathways. In contrast to IGF-I, leptin did not induce phosphorylation of IGF-IR, indicating that receptor cross-signaling is unidirectional, occurring from IGF-IR to Ob-R. Our results show, for the first time, a novel interaction and cross-talk between the IGF-I and leptin receptors in human breast cancer cells.

  6. Gene expression profiles of human dendritic cells interacting with Aspergillus fumigatus in a bilayer model of the alveolar epithelium/endothelium interface.

    Science.gov (United States)

    Morton, Charles Oliver; Fliesser, Mirjam; Dittrich, Marcus; Mueller, Tobias; Bauer, Ruth; Kneitz, Susanne; Hope, William; Rogers, Thomas Richard; Einsele, Hermann; Loeffler, Juergen

    2014-01-01

    The initial stages of the interaction between the host and Aspergillus fumigatus at the alveolar surface of the human lung are critical in the establishment of aspergillosis. Using an in vitro bilayer model of the alveolus, including both the epithelium (human lung adenocarcinoma epithelial cell line, A549) and endothelium (human pulmonary artery epithelial cells, HPAEC) on transwell membranes, it was possible to closely replicate the in vivo conditions. Two distinct sub-groups of dendritic cells (DC), monocyte-derived DC (moDC) and myeloid DC (mDC), were included in the model to examine immune responses to fungal infection at the alveolar surface. RNA in high quantity and quality was extracted from the cell layers on the transwell membrane to allow gene expression analysis using tailored custom-made microarrays, containing probes for 117 immune-relevant genes. This microarray data indicated minimal induction of immune gene expression in A549 alveolar epithelial cells in response to germ tubes of A. fumigatus. In contrast, the addition of DC to the system greatly increased the number of differentially expressed immune genes. moDC exhibited increased expression of genes including CLEC7A, CD209 and CCL18 in the absence of A. fumigatus compared to mDC. In the presence of A. fumigatus, both DC subgroups exhibited up-regulation of genes identified in previous studies as being associated with the exposure of DC to A. fumigatus and exhibiting chemotactic properties for neutrophils, including CXCL2, CXCL5, CCL20, and IL1B. This model closely approximated the human alveolus allowing for an analysis of the host pathogen interface that complements existing animal models of IA.

  7. Gene expression profiles of human dendritic cells interacting with Aspergillus fumigatus in a bilayer model of the alveolar epithelium/endothelium interface.

    Directory of Open Access Journals (Sweden)

    Charles Oliver Morton

    Full Text Available The initial stages of the interaction between the host and Aspergillus fumigatus at the alveolar surface of the human lung are critical in the establishment of aspergillosis. Using an in vitro bilayer model of the alveolus, including both the epithelium (human lung adenocarcinoma epithelial cell line, A549 and endothelium (human pulmonary artery epithelial cells, HPAEC on transwell membranes, it was possible to closely replicate the in vivo conditions. Two distinct sub-groups of dendritic cells (DC, monocyte-derived DC (moDC and myeloid DC (mDC, were included in the model to examine immune responses to fungal infection at the alveolar surface. RNA in high quantity and quality was extracted from the cell layers on the transwell membrane to allow gene expression analysis using tailored custom-made microarrays, containing probes for 117 immune-relevant genes. This microarray data indicated minimal induction of immune gene expression in A549 alveolar epithelial cells in response to germ tubes of A. fumigatus. In contrast, the addition of DC to the system greatly increased the number of differentially expressed immune genes. moDC exhibited increased expression of genes including CLEC7A, CD209 and CCL18 in the absence of A. fumigatus compared to mDC. In the presence of A. fumigatus, both DC subgroups exhibited up-regulation of genes identified in previous studies as being associated with the exposure of DC to A. fumigatus and exhibiting chemotactic properties for neutrophils, including CXCL2, CXCL5, CCL20, and IL1B. This model closely approximated the human alveolus allowing for an analysis of the host pathogen interface that complements existing animal models of IA.

  8. The Smart Factory

    DEFF Research Database (Denmark)

    Radziwon, Agnieszka; Bilberg, Arne; Bogers, Marcel

    2014-01-01

    Nowadays we live in a world, which a decade ago would only be described in the science fiction literature. More and more things become smart and both scientists and engineers strive for developing not only new and innovative devices, but also homes, factories, or even cities. Despite of continuous...... development, many of those concepts are still being just a vision of the future, which still needs a lot of effort to become true. This paper reviews the usage of adjective smart in respect to technology and with a special emphasis on the smart factory concept placement among contemporary studies. Due...... to a lack of a consensus of common understanding of this term, a unified definition is proposed. The conceptualization will not only refer to various smart factory visions reported in the literature, but also link the crucial characteristics of this emerging manufacturing concept to usual manufacturing...

  9. (+)-Geodin from Aspergillus terreus

    DEFF Research Database (Denmark)

    Rønnest, Mads Holger; Nielsen, Morten Thrane; Leber, Blanka

    2011-01-01

    The fungal metabolite (+)-geodin [systematic name: (2R)-methyl 5,7-dichloro-4-hydroxy-6'-methoxy-6-methyl-3,4'-dioxospiro[benzofuran-2,1'-cyclohexa-2',5'-diene]-2'-carboxylate], C(17)H(12)Cl(2)O(7), was isolated from Aspergillus terreus. The crystal structure contains two independent molecules...

  10. Production of biodiesel from plant oil hydrolysates using an Aspergillus oryzae whole-cell biocatalyst highly expressing Candida antarctica lipase B.

    Science.gov (United States)

    Adachi, Daisuke; Hama, Shinji; Nakashima, Kazunori; Bogaki, Takayuki; Ogino, Chiaki; Kondo, Akihiko

    2013-05-01

    For enzymatic biodiesel production from plant oil hydrolysates, an Aspergillus oryzae whole-cell biocatalyst that expresses Candida antarctica lipase B (r-CALB) with high esterification activity was developed. Each of soybean and palm oils was hydrolyzed using Candida rugosa lipase, and the resultant hydrolysates were subjected to esterification where immobilized r-CALB was used as a catalyst. In esterification, r-CALB afforded a methyl ester content of more than 90% after 6 h with the addition of 1.5 M equivalents of methanol. Favorably, stepwise additions of methanol and a little water were unnecessary for maintaining the lipase stability of r-CALB during esterification. During long-term esterification in a rotator, r-CALB can be recycled for 20 cycles without a significant loss of lipase activity, resulting in a methyl ester content of more than 90% even after the 20th batch. Therefore, the presented reaction system using r-CALB shows promise for biodiesel production from plant oil hydrolysates.

  11. Recognition of Aspergillus fumigatus hyphae by human plasmacytoid dendritic cells is mediated by dectin-2 and results in formation of extracellular traps.

    Directory of Open Access Journals (Sweden)

    Flávio V Loures

    2015-02-01

    Full Text Available Plasmacytoid dendritic cells (pDCs were initially considered as critical for innate immunity to viruses. However, our group has shown that pDCs bind to and inhibit the growth of Aspergillus fumigatus hyphae and that depletion of pDCs renders mice hypersusceptible to experimental aspergillosis. In this study, we examined pDC receptors contributing to hyphal recognition and downstream events in pDCs stimulated by A. fumigatus hyphae. Our data show that Dectin-2, but not Dectin-1, participates in A. fumigatus hyphal recognition, TNF-α and IFN-α release, and antifungal activity. Moreover, Dectin-2 acts in cooperation with the FcRγ chain to trigger signaling responses. In addition, using confocal and electron microscopy we demonstrated that the interaction between pDCs and A. fumigatus induced the formation of pDC extracellular traps (pETs containing DNA and citrullinated histone H3. These structures closely resembled those of neutrophil extracellular traps (NETs. The microarray analysis of the pDC transcriptome upon A. fumigatus infection also demonstrated up-regulated expression of genes associated with apoptosis as well as type I interferon-induced genes. Thus, human pDCs directly recognize A. fumigatus hyphae via Dectin-2; this interaction results in cytokine release and antifungal activity. Moreover, hyphal stimulation of pDCs triggers a distinct pattern of pDC gene expression and leads to pET formation.

  12. High efficiency cell-recycle continuous sodium gluconate production by Aspergillus niger using on-line physiological parameters association analysis to regulate feed rate rationally.

    Science.gov (United States)

    Lu, Fei; Li, Chao; Wang, Zejian; Zhao, Wei; Chu, Ju; Zhuang, Yingping; Zhang, Siliang

    2016-11-01

    In this paper, a system of cell-recycle continuous fermentation for sodium gluconate (SG) production by Aspergillus niger (A. niger) was established. Based on initial continuous fermentation result (100.0h) with constant feed rate, an automatic feedback strategy to regulate feed rate using on-line physiological parameters (OUR and DO) was proposed and applied successfully for the first time in the improved continuous fermentation (240.5h). Due to less auxiliary time, highest SG production rate (31.05±0.29gL(-1)h(-1)) and highest yield (0.984±0.067molmol(-1)), overall SG production capacity (975.8±5.8gh(-1)) in 50-L fermentor of improved continuous fermentation increased more than 300.0% compared to that of batch fermentation. Improvement of mass transfer and dispersed mycelia morphology were the two major reasons responsible for the high SG production rate. This system had been successfully applied to industrial fermentation and SG production was greatly improved.

  13. The Clone Factory

    Science.gov (United States)

    Stoddard, Beryl

    2005-01-01

    Have humans been cloned? Is it possible? Immediate interest is sparked when students are asked these questions. In response to their curiosity, the clone factory activity was developed to help them understand the process of cloning. In this activity, students reenact the cloning process, in a very simplified simulation. After completing the…

  14. Higgs Factory Concepts

    CERN Document Server

    Zimmermann, Frank

    2016-01-01

    Designs for future high-energy circular electron-positron colliders are based on both established and novel concepts. An appropriate design will enable these facilities to serve not only as “Higgs factories”, but also as Z, W and top factories, and, in addition, to become a possible first step to a higher-energy hadron collider.

  15. B-factory detectors

    CERN Document Server

    Marlow, D R

    2002-01-01

    The designs of the recently commissioned BaBar and Belle B-Factory detectors are described. The discussion is organized around the methods and instruments used to detect the so-called gold-plated-mode B sup 0->J/PSI K sub S decays and related modes.

  16. Transcription factories: genetic programming in three dimensions.

    Science.gov (United States)

    Edelman, Lucas Brandon; Fraser, Peter

    2012-04-01

    Among the most intensively studied systems in molecular biology is the eukaryotic transcriptional apparatus, which expresses genes in a regulated manner across hundreds of different cell types. Several studies over the past few years have added weight to the concept that transcription takes place within discrete 'transcription factories' assembled inside the cell nucleus. These studies apply innovative technical approaches to gain insights into the molecular constituents, dynamical behaviour and organizational regulators of transcription factories, providing exciting insights into the spatial dimension of transcriptional control.

  17. 2-hydroxyisocaproic acid is fungicidal for Candida and Aspergillus species.

    Science.gov (United States)

    Sakko, M; Moore, C; Novak-Frazer, L; Rautemaa, V; Sorsa, T; Hietala, P; Järvinen, A; Bowyer, P; Tjäderhane, L; Rautemaa, R

    2014-04-01

    The amino acid derivative 2-hydroxyisocaproic acid (HICA) is a nutritional additive used to increase muscle mass. Low levels can be detected in human plasma as a result of leucine metabolism. It has broad antibacterial activity but its efficacy against pathogenic fungi is not known. The aim was to test the efficacy of HICA against Candida and Aspergillus species. Efficacy of HICA against 19 clinical and reference isolates representing five Candida and three Aspergillus species with variable azole antifungal sensitivity profiles was tested using a microdilution method. The concentrations were 18, 36 and 72 mg ml(-1) . Growth was determined spectrophotometrically for Candida isolates and by visual inspection for Aspergillus isolates, viability was tested by culture and impact on morphology by microscopy. HICA of 72 mg ml(-1) was fungicidal against all Candida and Aspergillus fumigatus and Aspergillus terreus isolates. Lower concentrations were fungistatic. Aspergillus flavus was not inhibited by HICA. HICA inhibited hyphal formation in susceptible Candida albicans and A. fumigatus isolates and affected cell wall integrity. In conclusion, HICA has broad antifungal activity against Candida and Aspergillus at concentrations relevant for topical therapy. As a fungicidal agent with broad-spectrum bactericidal activity, it may be useful in the topical treatment of multispecies superficial infections.

  18. SWITCH: a dynamic CRISPR tool for genome engineering and metabolic pathway control for cell factory construction in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Garcia Vanegas, Katherina; Lehka, Beata Joanna; Mortensen, Uffe Hasbro

    2017-01-01

    to the pathway control state where production was optimized by downregulating an essential gene TSC13, hence, reducing formation of a byproduct. Conclusions We have successfully integrated two CRISPR tools, one for genetic engineering and one for pathway control, into one system and successfully used it for cell...

  19. Lactic acid bacteria as a cell factory for the delivery of functional biomolecules and ingredients in cereal-based beverages: a review.

    Science.gov (United States)

    Waters, Deborah M; Mauch, Alexander; Coffey, Aidan; Arendt, Elke K; Zannini, Emanuele

    2015-01-01

    In this review, we aim to describe the mechanisms by which LAB can fulfil the novel role of efficient cell factory for the production of functional biomolecules and food ingredients to enhance the quality of cereal-based beverages. LAB fermentation is a safe, economical, and traditional method of food preservation foremost, as well as having the additional benefits of flavor, texture, and nutrition amelioration. Additionally, LAB fermentation in known to render cereal-based foods and beverages safe, in a chemical-free, consumer-friendly manner, from an antinutrient and toxigenic perspective. Huge market opportunities and potential exist for food manufacturers who can provide the ideal functional beverage fulfilling consumer needs. Newly developed fermented cereal-based beverages must address markets globally including, high-nutrition markets (developing countries), lifestyle choice consumers (vegetarian, vegan, low-fat, low-salt, low-calorie), food-related non-communicable disease sufferers (cardiovascular disease, diabetes), and green label consumers (Western countries). To fulfil these recommendations, a suitable LAB starter culture and cereal-based raw materials must be developed. These strains would be suitable for the biopreservation of cereal beverages and, ideally, would be highly antifungal, anti-mycotoxigenic, mycotoxin-binding and proteolytic (neutralize toxic peptides and release flavor-contributing amino acids) with an ability to ferment cereals, whilst synthesizing oligosaccharides, thus presenting a major opportunity for the development of safe cereal-based prebiotic functional beverages to compete with and replace the existing dairy versions.

  20. Medical nutrition therapy: use of sourdough lactic acid bacteria as a cell factory for delivering functional biomolecules and food ingredients in gluten free bread.

    Science.gov (United States)

    Arendt, Elke K; Moroni, Alice; Zannini, Emanuele

    2011-08-30

    Celiac disease (CD) is an immune-mediated disease, triggered in genetically susceptible individuals by ingesting gluten from wheat, rye, barley, and other closely related cereal grains. Currently, the estimated prevalence of CD is around 1 % of the population in the western world and medical nutritional therapy (MNT) is the only accepted treatment for celiac disease. To date, the replacement of gluten in bread presents a significant technological challenge for the cereal scientist due to the low baking performance of gluten free products (GF). The increasing demand by the consumer for high quality gluten-free (GF) bread, clean labels and natural products is rising. Sourdough has been used since ancient times for the production of rye and wheat bread, its universal usage can be attributed to the improved quality, nutritional properties and shelf life of sourdough based breads. Consequently, the exploitation of sourdough for the production of GF breads appears tempting. This review will highlight how sourdough LAB can be an efficient cell factory for delivering functional biomolecules and food ingredients to enhance the quality of gluten free bread.

  1. Medical nutrition therapy: use of sourdough lactic acid bacteria as a cell factory for delivering functional biomolecules and food ingredients in gluten free bread

    LENUS (Irish Health Repository)

    2011-08-30

    Abstract Celiac disease (CD) is an immune-mediated disease, triggered in genetically susceptible individuals by ingesting gluten from wheat, rye, barley, and other closely related cereal grains. Currently, the estimated prevalence of CD is around 1 % of the population in the western world and medical nutritional therapy (MNT) is the only accepted treatment for celiac disease. To date, the replacement of gluten in bread presents a significant technological challenge for the cereal scientist due to the low baking performance of gluten free products (GF). The increasing demand by the consumer for high quality gluten-free (GF) bread, clean labels and natural products is rising. Sourdough has been used since ancient times for the production of rye and wheat bread, its universal usage can be attributed to the improved quality, nutritional properties and shelf life of sourdough based breads. Consequently, the exploitation of sourdough for the production of GF breads appears tempting. This review will highlight how sourdough LAB can be an efficient cell factory for delivering functional biomolecules and food ingredients to enhance the quality of gluten free bread.

  2. Medical nutrition therapy: use of sourdough lactic acid bacteria as a cell factory for delivering functional biomolecules and food ingredients in gluten free bread

    Directory of Open Access Journals (Sweden)

    Moroni Alice

    2011-08-01

    Full Text Available Abstract Celiac disease (CD is an immune-mediated disease, triggered in genetically susceptible individuals by ingesting gluten from wheat, rye, barley, and other closely related cereal grains. Currently, the estimated prevalence of CD is around 1 % of the population in the western world and medical nutritional therapy (MNT is the only accepted treatment for celiac disease. To date, the replacement of gluten in bread presents a significant technological challenge for the cereal scientist due to the low baking performance of gluten free products (GF. The increasing demand by the consumer for high quality gluten-free (GF bread, clean labels and natural products is rising. Sourdough has been used since ancient times for the production of rye and wheat bread, its universal usage can be attributed to the improved quality, nutritional properties and shelf life of sourdough based breads. Consequently, the exploitation of sourdough for the production of GF breads appears tempting. This review will highlight how sourdough LAB can be an efficient cell factory for delivering functional biomolecules and food ingredients to enhance the quality of gluten free bread.

  3. Elicitation, an Effective Strategy for the Biotechnological Production of Bioactive High-Added Value Compounds in Plant Cell Factories

    Directory of Open Access Journals (Sweden)

    Karla Ramirez-Estrada

    2016-02-01

    Full Text Available Plant in vitro cultures represent an attractive and cost-effective alternative to classical approaches to plant secondary metabolite (PSM production (the “Plant Cell Factory” concept. Among other advantages, they constitute the only sustainable and eco-friendly system to obtain complex chemical structures biosynthesized by rare or endangered plant species that resist domestication. For successful results, the biotechnological production of PSM requires an optimized system, for which elicitation has proved one of the most effective strategies. In plant cell cultures, an elicitor can be defined as a compound introduced in small concentrations to a living system to promote the biosynthesis of the target metabolite. Traditionally, elicitors have been classified in two types, abiotic or biotic, according to their chemical nature and exogenous or endogenous origin, and notably include yeast extract, methyl jasmonate, salicylic acid, vanadyl sulphate and chitosan. In this review, we summarize the enhancing effects of elicitors on the production of high-added value plant compounds such as taxanes, ginsenosides, aryltetralin lignans and other types of polyphenols, focusing particularly on the use of a new generation of elicitors such as coronatine and cyclodextrins.

  4. Chemodiversity in the genus Aspergillus.

    Science.gov (United States)

    Frisvad, Jens C; Larsen, Thomas O

    2015-10-01

    Isolates of Aspergillus species are able to produce a large number of secondary metabolites. The profiles of biosynthetic families of secondary metabolites are species specific, whereas individual secondary metabolite families can occur in other species, even those phylogenetically and ecologically unrelated to Aspergillus. Furthermore, there is a high degree of chemo-consistency from isolate to isolate in a species even though certain metabolite gene clusters are silenced in some isolates. Genome sequencing projects have shown that the diversity of secondary metabolites is much larger in each species than previously thought. The potential of finding even further new bioactive drug candidates in Aspergillus is evident, despite the fact that many secondary metabolites have already been structure elucidated and chemotaxonomic studies have shown that many new secondary metabolites have yet to be characterized. The genus Aspergillus is cladistically holophyletic but phenotypically polythetic and very diverse and is associated to quite different sexual states. Following the one fungus one name system, the genus Aspergillus is restricted to a holophyletic clade that include the morphologically different genera Aspergillus, Dichotomomyces, Phialosimplex, Polypaecilum and Cristaspora. Secondary metabolites common between the subgenera and sections of Aspergillus are surprisingly few, but many metabolites are common to a majority of species within the sections. We call small molecule extrolites in the same biosynthetic family isoextrolites. However, it appears that secondary metabolites from one Aspergillus section have analogous metabolites in other sections (here also called heteroisoextrolites). In this review, we give a genus-wide overview of secondary metabolite production in Aspergillus species. Extrolites appear to have evolved because of ecological challenges rather than being inherited from ancestral species, at least when comparing the species in the different

  5. The Super Flavor Factory

    Energy Technology Data Exchange (ETDEWEB)

    Bevan, A.J.; /Queen Mary, U. of London

    2007-01-26

    The main physics goals of a high luminosity e{sup +}e{sup -} flavor factory are discussed, including the possibilities to perform detailed studies of the CKM mechanism of quark mixing, and constrain virtual Higgs and Non-Standard Model particle contributions to the dynamics of rare B{sub u,d,s} decays. The large samples of D mesons and {tau} leptons produced at a flavor factory will result in improved sensitivities on D mixing and lepton flavor violation searches, respectively. One can also test fundamental concepts such as lepton universality to much greater precision than existing constraints and improve the precision on tests of CPT from B meson decays. Recent developments in accelerator physics have demonstrated the feasibility to build an accelerator that can achieve luminosities of {Omicron}(10{sup 36} cm{sup -2} s{sup -1}).

  6. Engineering the smart factory

    Science.gov (United States)

    Harrison, Robert; Vera, Daniel; Ahmad, Bilal

    2016-10-01

    The fourth industrial revolution promises to create what has been called the smart factory. The vision is that within such modular structured smart factories, cyber-physical systems monitor physical processes, create a virtual copy of the physical world and make decentralised decisions. This paper provides a view of this initiative from an automation systems perspective. In this context it considers how future automation systems might be effectively configured and supported through their lifecycles and how integration, application modelling, visualisation and reuse of such systems might be best achieved. The paper briefly describes limitations in current engineering methods, and new emerging approaches including the cyber physical systems (CPS) engineering tools being developed by the automation systems group (ASG) at Warwick Manufacturing Group, University of Warwick, UK.

  7. Saccharomyces pastorianus as cell factory to improve production of fructose 1,6-diphosphate using novel fermentation strategies

    Directory of Open Access Journals (Sweden)

    Chiara Schiraldi

    2015-08-01

    Full Text Available Enzymatic phosphorylation of glucose with inorganic phosphate, mediated by permeabilized yeast cells, is one of the methods commonly used to manufacture fructose 1,6-diphosphate, a compound of pharmaceutical interest. This process requires high concentrations of yeast active biomass, that is the catalyst of bioconversion of glucose and inorganic phosphate into fructose 1,6-diphosphate. In this study we firstly describe the high cell density production of a brewer's Saccharomyces strain (Saccharomyces pastorianus DSM 6581, focusing on the optimization of medium composition and exploiting fed-batch strategies and novel technologies based on membrane bioreactors. In fed-batch fermentation an appropriate exponential feed profile was set up to maintain the glucose concentration in the bioreactor below 0.9 g·L-1, thus yielding reproducibly 58 g dry weight biomass per liter in 80 h fermentation, improving eight-fold batch processes output. In addition a higher final biomass density was reached when implementing a microfiltration strategy (70 g dry weight biomass, that led to a productivity of 2.1 gcdw·L-1·h-1, 2.4-fold the fed-batch one. Successively, this biomass was opportunely permeabilized and proved capable of catalyzing the bioconversion of glucose into fructose 1,6-diphosphate. Acting on critical parameters of the bioconversion (substrates molar ratio, catalyst concentration and permeabilization agent, fructose 1,6-diphosphate was produced, after 3 h of process, at 56.3 ± 1 g·L-1 with a yield of 80% of the theoretical value.

  8. Maximum Autocorrelation Factorial Kriging

    OpenAIRE

    Nielsen, Allan Aasbjerg; Conradsen, Knut; Pedersen, John L.; Steenfelt, Agnete

    2000-01-01

    This paper describes maximum autocorrelation factor (MAF) analysis, maximum autocorrelation factorial kriging, and its application to irregularly sampled stream sediment geochemical data from South Greenland. Kriged MAF images are compared with kriged images of varimax rotated factors from an ordinary non-spatial factor analysis, and they are interpreted in a geological context. It is demonstrated that MAF analysis contrary to ordinary non-spatial factor analysis gives an objective discrimina...

  9. Analysis of Factorial Experiments.

    Science.gov (United States)

    1985-09-01

    The pro- bability that y 13 is bad is .37. Choosing Q greater than .37 ends the iteration. Choos - ing Q so that Y 13 is identified as a bad...ances as a Fraction of Their Total. Annals of Eugenics , 11, 47-52. Cochran, W. G. and Cox, G. M. (1957) Experimental Designs. New York: John Wiley...Experiments. London: Oliver and Boyd. Finney, D. J. (1945) The Fractional Replication of Factorial Arrangements. Annals of Eugenics , 12, 291-301

  10. Phi factory detector requirements

    Energy Technology Data Exchange (ETDEWEB)

    Arisaka, K.; Atac, M.; Berg, R.; Buchanan, C.; Calvette, M.; Khazin, B.; Kinoshita, K.; Muller, T.; Ohshima, T.; Olsen, S.; Park, J.; Santoni, C.; Shirai, J.; Solodov, E.; Thompson, J.; Triggiani, G.; Ueno, K.; Yamamoto, H.; Detector and Simulation Working Group

    1991-08-01

    We identify the experimental problems and the conditions required for successful phi-factory operation, and show the range of detector parameters which, in conjunction with different machine designs, may meet these conditions. We started by considering, comparing and criticizing the Italian and Novosibirsk designs. With this discussion as a background, we defined the apparent experimental problems and detector constraints. In this article we summarize our understanding. (orig./HSI).

  11. The transcriptional repressor TupA in Aspergillus niger is involved in controlling gene expression related to cell wall biosynthesis, development, and nitrogen source availability.

    Directory of Open Access Journals (Sweden)

    Doreen Schachtschabel

    Full Text Available The Tup1-Cyc8 (Ssn6 complex is a well characterized and conserved general transcriptional repressor complex in eukaryotic cells. Here, we report the identification of the Tup1 (TupA homolog in the filamentous fungus Aspergillus niger in a genetic screen for mutants with a constitutive expression of the agsA gene. The agsA gene encodes a putative alpha-glucan synthase, which is induced in response to cell wall stress in A. niger. Apart from the constitutive expression of agsA, the selected mutant was also found to produce an unknown pigment at high temperatures. Complementation analysis with a genomic library showed that the tupA gene could complement the phenotypes of the mutant. Screening of a collection of 240 mutants with constitutive expression of agsA identified sixteen additional pigment-secreting mutants, which were all mutated in the tupA gene. The phenotypes of the tupA mutants were very similar to the phenotypes of a tupA deletion strain. Further analysis of the tupA-17 mutant and the ΔtupA mutant revealed that TupA is also required for normal growth and morphogenesis. The production of the pigment at 37°C is nitrogen source-dependent and repressed by ammonium. Genome-wide expression analysis of the tupA mutant during exponential growth revealed derepression of a large group of diverse genes, including genes related to development and cell wall biosynthesis, and also protease-encoding genes that are normally repressed by ammonium. Comparison of the transcriptome of up-regulated genes in the tupA mutant showed limited overlap with the transcriptome of caspofungin-induced cell wall stress-related genes, suggesting that TupA is not a general suppressor of cell wall stress-induced genes. We propose that TupA is an important repressor of genes related to development and nitrogen metabolism.

  12. Deletion of flbA results in increased secretome complexity and reduced secretion heterogeneity in colonies of Aspergillus niger.

    Science.gov (United States)

    Krijgsheld, Pauline; Nitsche, Benjamin M; Post, Harm; Levin, Ana M; Müller, Wally H; Heck, Albert J R; Ram, Arthur F J; Altelaar, A F Maarten; Wösten, Han A B

    2013-04-05

    Aspergillus niger is a cell factory for the production of enzymes. This fungus secretes proteins in the central part and at the periphery of the colony. The sporulating zone of the colony overlapped with the nonsecreting subperipheral zone, indicating that sporulation inhibits protein secretion. Indeed, strain ΔflbA that is affected early in the sporulation program secreted proteins throughout the colony. In contrast, the ΔbrlA strain that initiates but not completes sporulation did not show altered spatial secretion. The secretome of 5 concentric zones of xylose-grown ΔflbA colonies was assessed by quantitative proteomics. In total 138 proteins with a signal sequence for secretion were identified in the medium of ΔflbA colonies. Of these, 18 proteins had never been reported to be part of the secretome of A. niger, while 101 proteins had previously not been identified in the culture medium of xylose-grown wild type colonies. Taken together, inactivation of flbA results in spatial changes in secretion and in a more complex secretome. The latter may be explained by the fact that strain ΔflbA has a thinner cell wall compared to the wild type, enabling efficient release of proteins. These results are of interest to improve A. niger as a cell factory.

  13. Putative PmrA and PmcA are important for normal growth, morphogenesis and cell wall integrity, but not for viability in Aspergillus nidulans.

    Science.gov (United States)

    Jiang, Hechun; Liu, Feifei; Zhang, Shizhu; Lu, Ling

    2014-11-01

    P-type Ca(2+)-transporting ATPases are Ca(2+) pumps, extruding cytosolic Ca(2+) to the extracellular environment or the intracellular Ca(2+) store lumens. In budding yeast, Pmr1 (plasma membrane ATPase related), and Pmc1 (plasma membrane calcium-ATPase) cannot be deleted simultaneously for it to survive in standard medium. Here, we deleted two putative Ca(2+) pumps, designated AnPmrA and AnPmcA, from Aspergillus nidulans, and obtained the mutants ΔanpmrA and ΔanpmcA, respectively. Then, using ΔanpmrA as the starting strain, the promoter of its anpmcA was replaced with the alcA promoter to secure the mutant ΔanpmrAalcApmcA or its anpmcA was deleted completely to produce the mutant ΔanpmrAΔpmcA. Different from the case in Saccharomyces cerevisiae, double deletion of anpmrA and anpmcA was not lethal in A. nidulans. In addition, deletion of anpmrA and/or anpmcA had produced growth defects, although overexpression of AnPmc1 in ΔanpmrAalcApmcA could not restore the growth defects that resulted from the loss of AnPmrA. Moreover, we found AnPmrA was indispensable for maintenance of normal morphogenesis, especially in low-Ca(2+)/Mn(2+) environments. Thus, our findings suggest AnPmrA and AnPmcA might play important roles in growth, morphogenesis and cell wall integrity in A. nidulans in a different way from that in yeasts.

  14. A trispecies Aspergillus microarray: Comparative transcriptomics of three Aspergillus species

    DEFF Research Database (Denmark)

    Andersen, Mikael Rørdam; Vongsangnak, Wanwipa; Panagiotou, Gianni

    2008-01-01

    The full-genome sequencing of the filamentous fungi Aspergillus nidulans, Aspergillus niger, and Aspergillus oryzae has opened possibilities for studying the cellular physiology of these fungi on a systemic level. As a tool to explore this, we are making available an Affymetrix GeneChip developed...... data identified 23 genes to be a conserved response across Aspergillus sp., including the xylose transcriptional activator XlnR. A promoter analysis of the up-regulated genes in all three species indicates the conserved XInR-binding site to be 5'-GGNTAAA-3'. The composition of the conserved gene......-set suggests that xylose acts as a molecule, indicating the presence of complex carbohydrates such as hemicellulose, and triggers an array of degrading enzymes. With this case example, we present a validated tool for transcriptome analysis of three Aspergillus species and a methodology for conducting cross...

  15. Diversity of Aspergillus oryzae genotypes (RFLP) isolated from traditional soy sauce production within Malaysia and Southeast Asia

    Science.gov (United States)

    DNA fingerprinting was performed on 64 strains of Aspergillus oryzae and one strain of A. sojae isolated from soysauce factories within Malaysia and Southeast Asia that use primitive traditional methods in producing 'tamari type' Cantonese soy sauce. PstI digests of total genomic DNA from each isol...

  16. Deciding WQO for factorial languages

    KAUST Repository

    Atminas, Aistis

    2013-04-05

    A language is factorial if it is closed under taking factors (i.e. contiguous subwords). Every factorial language can be described by an antidictionary, i.e. a minimal set of forbidden factors. We show that the problem of deciding whether a factorial language given by a finite antidictionary is well-quasi-ordered under the factor containment relation can be solved in polynomial time. © 2013 Springer-Verlag Berlin Heidelberg.

  17. Aspergillus saccharolyticus sp. nov., a new black Aspergillus species isolated in Denmark

    DEFF Research Database (Denmark)

    Sørensen, Annette; Lübeck, Peter S.; Lübeck, Mette

    2011-01-01

    A novel species, Aspergillus saccharolyticus sp. nov., belonging to the Aspergillus section Nigri group is described. This species was isolated in Denmark from treated hardwood. Its taxonomic status was determined using a polyphasic taxonomic approach including phenotypic (morphology and extrolite...... Aspergillus species that is morphologically similar to Aspergillus japonicus and Aspergillus aculeatus, but has a totally different extrolite profile compared to any known Aspergillus species. The type strain of A. saccharolyticus sp. nov. is CBS 127449T ( = IBT 28509T)....

  18. Effect of gamma radiation on Aspergillus flavus and Aspergillus ochraceus ultrastructure and mycotoxin production

    Science.gov (United States)

    Ribeiro, J.; Cavaglieri, L.; Vital, H.; Cristofolini, A.; Merkis, C.; Astoreca, A.; Orlando, J.; Carú, M.; Dalcero, A.; Rosa, C. A. R.

    2011-05-01

    The aim of this work was to study the effect of gamma radiation (2 kGy) on Aspergillus flavus and Aspergillus ochraceus ultrastructure. Moreover, the influence on aflatoxin B 1 and ochratoxin A production was also observed. Irradiated A. flavus strain showed a dull orangish colony while control strain showed the typical green color. Minor differences were observed on stipes, metulae and conidia size between control and irradiated A. flavus and A. ochraceus strains. Irradiated fungi showed ultrastructural changes on cell wall, plasmalema and cytoplasm levels. The levels of mycotoxins produced by irradiated strains were two times greater than those produced by control strains. Successive transferences of irradiated strains on malt extract agar allowed the fungus to recuperate morphological characteristics. Although minor changes in the fungal morphology were observed, ultrastructural changes at cell wall level and the increase of mycotoxin production ability were observed. Inappropriate storage of irradiated food and feed would allow the development of potentially more toxicogenic fungal propagules.

  19. Micro Factory Systems and Tooling

    DEFF Research Database (Denmark)

    Hansen, Hans Nørgaard

    2003-01-01

    The presentation deals with the micro factory concept in general and with the issues related to manufacturing, handling/assembly and quality control in particular.......The presentation deals with the micro factory concept in general and with the issues related to manufacturing, handling/assembly and quality control in particular....

  20. Replicating DNA by cell factories: roles of central carbon metabolism and transcription in the control of DNA replication in microbes, and implications for understanding this process in human cells.

    Science.gov (United States)

    Barańska, Sylwia; Glinkowska, Monika; Herman-Antosiewicz, Anna; Maciąg-Dorszyńska, Monika; Nowicki, Dariusz; Szalewska-Pałasz, Agnieszka; Węgrzyn, Alicja; Węgrzyn, Grzegorz

    2013-05-29

    Precise regulation of DNA replication is necessary to ensure the inheritance of genetic features by daughter cells after each cell division. Therefore, determining how the regulatory processes operate to control DNA replication is crucial to our understanding and application to biotechnological processes. Contrary to early concepts of DNA replication, it appears that this process is operated by large, stationary nucleoprotein complexes, called replication factories, rather than by single enzymes trafficking along template molecules. Recent discoveries indicated that in bacterial cells two processes, central carbon metabolism (CCM) and transcription, significantly and specifically influence the control of DNA replication of various replicons. The impact of these discoveries on our understanding of the regulation of DNA synthesis is discussed in this review. It appears that CCM may influence DNA replication by either action of specific metabolites or moonlighting activities of some enzymes involved in this metabolic pathway. The role of transcription in the control of DNA replication may arise from either topological changes in nucleic acids which accompany RNA synthesis or direct interactions between replication and transcription machineries. Due to intriguing similarities between some prokaryotic and eukaryotic regulatory systems, possible implications of studies on regulation of microbial DNA replication on understanding such a process occurring in human cells are discussed.

  1. Aspergillus fumigatus in Poultry

    Directory of Open Access Journals (Sweden)

    Pascal Arné

    2011-01-01

    Full Text Available Aspergillus fumigatus remains a major respiratory pathogen in birds. In poultry, infection by A. fumigatus may induce significant economic losses particularly in turkey production. A. fumigatus develops and sporulates easily in poor quality bedding or contaminated feedstuffs in indoor farm environments. Inadequate ventilation and dusty conditions increase the risk of bird exposure to aerosolized spores. Acute cases are seen in young animals following inhalation of spores, causing high morbidity and mortality. The chronic form affects older birds and looks more sporadic. The respiratory tract is the primary site of A. fumigatus development leading to severe respiratory distress and associated granulomatous airsacculitis and pneumonia. Treatments for infected poultry are nonexistent; therefore, prevention is the only way to protect poultry. Development of avian models of aspergillosis may improve our understanding of its pathogenesis, which remains poorly understood.

  2. Aspergillus-Related Lung Disease

    Directory of Open Access Journals (Sweden)

    Alia Al-Alawi

    2005-01-01

    Full Text Available Aspergillus is a ubiquitous dimorphic fungus that causes a variety of human diseases ranging in severity from trivial to life-threatening, depending on the host response. An intact host defence is important to prevent disease, but individuals with pre-existing structural lung disease, atopy, occupational exposure or impaired immunity are susceptible. Three distinctive patterns of aspergillus-related lung disease are recognized: saprophytic infestation of airways, cavities and necrotic tissue; allergic disease including extrinsic allergic alveolitis, asthma, allergic bronchopulmonary aspergillosis, bronchocentric granulomatosis and chronic eosinophilic pneumonia; and airway and tissue invasive disease -- pseudomembranous tracheobronchitis, acute bronchopneumonia, angioinvasive aspergillosis, chronic necrotizing aspergillosis and invasive pleural disease. A broad knowledge of these clinical presentations and a high index of suspicion are required to ensure timely diagnosis and treatment of the potentially lethal manifestations of aspergillus-related pulmonary disease. In the present report, the clinical, radiographic and pathological aspects of the various aspergillus-related lung diseases are briefly reviewed.

  3. Aspergillus antigen skin test (image)

    Science.gov (United States)

    ... After 48 to 72 hours the site of injection is evaluated by a physician. If a positive reaction occurs (the test site is inflamed), the person has been exposed to the aspergillus mold and is at risk for developing aspergillosis.

  4. 76 FR 16297 - Aspergillus flavus

    Science.gov (United States)

    2011-03-23

    ..., lawns, or buildings (residential and other indoor uses). A. Dietary Exposure 1. Food. Current uses of... emergence). Once applied to corn and after exposure to moisture, Aspergillus flavus AF36 germinates,...

  5. The Nearby Supernova Factory

    CERN Document Server

    Wood-Vasey, W M; Lee Byung Cheol; Loken, S; Nugent, P; Perlmutter, S; Siegrist, J L; Wang, L; Antilogus, P; Astier, Pierre; Hardin, D; Pain, R; Copin, Y; Smadja, G; Gangler, E; Castera, A; Adam, G; Bacon, R; Lemonnier, J P; Pecontal, A; Pécontal, E; Kessler, R

    2004-01-01

    The Nearby Supernova Factory (SNfactory) is an ambitious project to find and study in detail approximately 300 nearby Type Ia supernovae (SNe~Ia) at redshifts 0.03

  6. AutoPyFactory: A Scalable Flexible Pilot Factory Implementation

    Science.gov (United States)

    Caballero, J.; Hover, J.; Love, P.; Stewart, G. A.

    2012-12-01

    The ATLAS experiment at the CERN LHC is one of the largest users of grid computing infrastructure, which is a central part of the experiment's computing operations. Considerable efforts have been made to use grid technology in the most efficient and effective way, including the use of a pilot job based workload management framework. In this model the experiment submits ‘pilot’ jobs to sites without payload. When these jobs begin to run they contact a central service to pick-up a real payload to execute. The first generation of pilot factories were usually specific to a single Virtual Organization (VO), and were bound to the particular architecture of that VO's distributed processing. A second generation provides factories which are more flexible, not tied to any particular VO, and provide new and improved features such as monitoring, logging, profiling, etc. In this paper we describe this key part of the ATLAS pilot architecture, a second generation pilot factory, AutoPyFactory. AutoPyFactory has a modular design and is highly configurable. It is able to send different types of pilots to sites and exploit different submission mechanisms and queue characteristics. It is tightly integrated with the PanDA job submission framework, coupling pilot flow to the amount of work the site has to run. It gathers information from many sources in order to correctly configure itself for a site and its decision logic can easily be updated. Integrated into AutoPyFactory is a flexible system for delivering both generic and specific job wrappers which can perform many useful actions before starting to run end-user scientific applications, e.g., validation of the middleware, node profiling and diagnostics, and monitoring. AutoPyFactory also has a robust monitoring system that has been invaluable in establishing a reliable pilot factory service for ATLAS.

  7. Muon colliders and neutrino factories

    CERN Document Server

    Geer, S

    2012-01-01

    Over the last decade there has been significant progress in developing the concepts and technologies needed to produce, capture and accelerate O(1021) muons/year. This development prepares the way for a new type of neutrino source (Neutrino Factory) and a new type of very high energy lepton-antilepton collider (Muon Collider). This article reviews the motivation, design and R&D for Neutrino Factories and Muon Colliders.

  8. Progress in Research on the Methylotrophic Yeast Hansenula polymaorph Cell Factory%多形汉逊酵母作为细胞工厂的应用研究进展

    Institute of Scientific and Technical Information of China (English)

    钱卫东; 施春阳; 王婷

    2012-01-01

    The thermotolerant methylotrophic yeast Hansenula polymarpha has been an important cell factory for the production of pharmaceutical proteins and enzyme preparation, but recently been also gaining interest as a promising host for the production of both bioethanol and efficacious components from medicinal plant. In this paper, some characteristics of H. poly-morpha were outlined,and the progress in research on H. polymorpha as a microbiology cell factory was summarized. Finally, the prospective application of H. polymorpha to biotechnology research was discussed.%多形汉逊酵母以其独特的生物学和遗传学特征已成为一种重要的细胞工厂,被广泛运用于生产药物蛋白、酶制剂、生物能源及药用植物有效成分等.作者概述了多形汉逊酵母的一些基本特性,阐述了其作为微生物细胞工厂的应用研究进展,并对其未来工作的前景进行了展望.

  9. Disruption of the Phospholipase D Gene Attenuates the Virulence of Aspergillus fumigatus

    NARCIS (Netherlands)

    Li, Xianping; Gao, Meihua; Han, Xuelin; Tao, Sha; Zheng, Dongyu; Cheng, Ying; Yu, Rentao; Han, Gaige; Schmidt, Martina; Han, Li

    2012-01-01

    Aspergillus fumigatus is the most prevalent airborne fungal pathogen that induces serious infections in immunocompromised patients. Phospholipases are key enzymes in pathogenic fungi that cleave host phospholipids, resulting in membrane destabilization and host cell penetration. However, knowledge o

  10. Anti-Inflammatory and Cytoprotective Effects of TMC-256C1 from Marine-Derived Fungus Aspergillus sp. SF-6354 via up-Regulation of Heme Oxygenase-1 in Murine Hippocampal and Microglial Cell Lines

    Directory of Open Access Journals (Sweden)

    Dong-Cheol Kim

    2016-04-01

    Full Text Available In the course of searching for bioactive secondary metabolites from marine fungi, TMC-256C1 was isolated from an ethyl acetate extract of the marine-derived fungus Aspergillus sp. SF6354. TMC-256C1 displayed anti-neuroinflammatory effect in BV2 microglial cells induced by lipopolysaccharides (LPS as well as neuroprotective effect against glutamate-stimulated neurotoxicity in mouse hippocampal HT22 cells. TMC-256C1 was shown to develop a cellular resistance to oxidative damage caused by glutamate-induced cytotoxicity and reactive oxygen species (ROS generation in HT22 cells, and suppress the inflammation process in LPS-stimulated BV2 cells. Furthermore, the neuroprotective and anti-neuroinflammatory activities of TMC-256C1 were associated with upregulated expression of heme oxygenase (HO-1 and nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2 in HT22 and BV2 cells. We also found that TMC-256C1 activated p38 mitogen-activated protein kinases (MAPK and phosphatidylinositol 3-kinase (PI3K/Akt signaling pathways in HT22 and BV2 cells. These results demonstrated that TMC-256C1 activates HO-1 protein expression, probably by increasing nuclear Nrf2 levels via the activation of the p38 MAPK and PI3K/Akt pathways.

  11. Pyrophen Produced by Endophytic Fungi Aspergillus sp Isolated from Piper crocatum Ruiz and Pav Exhibits Cytotoxic Activity and Induces S Phase Arrest in T47D Breast Cancer Cells.

    Science.gov (United States)

    Astuti, Puji; Erden, Willy; Wahyono; Wahyuono, Subagus; Hertiani, Triana

    2016-01-01

    Ethyl acetate extracts obtained from culture of endophytic fungi Aspergillus sp isolated from Piper crocatum Ruiz and Pav, have been shown to possess cytotoxic activity against T47D breast cancer cells. Investigations were here conducted to determine bioactive compounds responsible for the activity. Bioassay guided fractionation was employed to obtain active compounds. Structure elucidation was performed based on analysis of LC-MS, 1H-NMR, 13C-NMR, COSY, DEPT, HMQC, HMBC data. Cytotoxity assays were conducted in 96 well plates against T47D and Vero cell lines. Bioassay guided isolation and chemical investigation led to the isolation of pyrophen, a 4-methoxy-6-(1'-acetamido-2'-phenylethyl)-2H-pyran-2-one. Further analysis of its activity against T47D and Vero cells showed an ability to inhibit the growth of T47D cells with IC50 values of 9.2 μg/mL but less cytotoxicity to Vero cells with an IC50 of 109 μg/mL. This compound at a concentration of 400 ng/mL induced S-phase arrest in T47D cells.

  12. The evolutionary imprint of domestication on genome variation and function of the filamentous fungus Aspergillus oryzae.

    Science.gov (United States)

    Gibbons, John G; Salichos, Leonidas; Slot, Jason C; Rinker, David C; McGary, Kriston L; King, Jonas G; Klich, Maren A; Tabb, David L; McDonald, W Hayes; Rokas, Antonis

    2012-08-01

    The domestication of animals, plants, and microbes fundamentally transformed the lifestyle and demography of the human species [1]. Although the genetic and functional underpinnings of animal and plant domestication are well understood, little is known about microbe domestication [2-6]. Here, we systematically examined genome-wide sequence and functional variation between the domesticated fungus Aspergillus oryzae, whose saccharification abilities humans have harnessed for thousands of years to produce sake, soy sauce, and miso from starch-rich grains, and its wild relative A. flavus, a potentially toxigenic plant and animal pathogen [7]. We discovered dramatic changes in the sequence variation and abundance profiles of genes and wholesale primary and secondary metabolic pathways between domesticated and wild relative isolates during growth on rice. Our data suggest that, through selection by humans, an atoxigenic lineage of A. flavus gradually evolved into a "cell factory" for enzymes and metabolites involved in the saccharification process. These results suggest that whereas animal and plant domestication was largely driven by Neolithic "genetic tinkering" of developmental pathways, microbe domestication was driven by extensive remodeling of metabolism.

  13. Improving the Secretory Expression of an α-Galactosidase from Aspergillus niger in Pichia pastoris

    Science.gov (United States)

    Zheng, Xianliang; Fang, Bo; Han, Dongfei; Yang, Wenxia; Qi, Feifei; Chen, Hui; Li, Shengying

    2016-01-01

    α-Galactosidases are broadly used in feed, food, chemical, pulp, and pharmaceutical industries. However, there lacks a satisfactory microbial cell factory that is able to produce α-galactosidases efficiently and cost-effectively to date, which prevents these important enzymes from greater application. In this study, the secretory expression of an Aspergillus niger α-galactosidase (AGA) in Pichia pastoris was systematically investigated. Through codon optimization, signal peptide replacement, comparative selection of host strain, and saturation mutagenesis of the P1’ residue of Kex2 protease cleavage site for efficient signal peptide removal, a mutant P. pastoris KM71H (Muts) strain of AGA-I with the specific P1’ site substitution (Glu to Ile) demonstrated remarkable extracellular α-galactosidase activity of 1299 U/ml upon a 72 h methanol induction in 2.0 L fermenter. The engineered yeast strain AGA-I demonstrated approximately 12-fold higher extracellular activity compared to the initial P. pastoris strain. To the best of our knowledge, this represents the highest yield and productivity of a secreted α-galactosidase in P. pastoris, thus holding great potential for industrial application. PMID:27548309

  14. The Aspergillus Mine - publishing bioinformatics

    DEFF Research Database (Denmark)

    Vesth, Tammi Camilla; Rasmussen, Jane Lind Nybo; Theobald, Sebastian

    so with no computational specialist. Here we present a setup for analysis and publication of genome data of 70 species of Aspergillus fungi. The platform is based on R, Python and uses the RShiny framework to create interactive web‐applications. It allows all participants to create interactive...... analysis which can be shared with the team and in connection with publications. We present analysis for investigation of genetic diversity, secondary and primary metabolism and general data overview. The platform, the Aspergillus Mine, is a collection of analysis tools based on data from collaboration...... with the Joint Genome Institute. The Aspergillus Mine is not intended as a genomic data sharing service but instead focuses on creating an environment where the results of bioinformatic analysis is made available for inspection. The data and code is public upon request and figures can be obtained directly from...

  15. Regulatory processes in Aspergillus niger

    DEFF Research Database (Denmark)

    Poulsen, Lars

    Filamentous fungi are extensively used in the fermentation industry for synthesis of numerous products. One of the most important, is the fungus Aspergillus niger, used industrially for production of organic acids, and homologous as well as heterologous enzymes. This fungus has numerous of advant......Filamentous fungi are extensively used in the fermentation industry for synthesis of numerous products. One of the most important, is the fungus Aspergillus niger, used industrially for production of organic acids, and homologous as well as heterologous enzymes. This fungus has numerous...

  16. Sustained low-dose growth hormone therapy optimizes bioactive insulin-like growth factor-I level and may enhance CD4 T-cell number in HIV infection

    DEFF Research Database (Denmark)

    Andersen, Ove; Hansen, Birgitte Rønde; Troensegaard, William;

    2010-01-01

    High-dose recombinant human growth hormone (rhGH) (2-6 mg/day) regimes may facilitate T-cell restoration in patients infected with human immunodeficiency virus (HIV) on highly active antiretroviral therapy (HAART). However, high-dose rhGH regimens increase insulin-like growth factor-I (IGF......-I) to supra-physiological levels associated with severe side effects. The present study investigated whether lower doses of rhGH may improve T-cell restoration in patients infected with HIV following an expedient response of total and bioactive (i.e., free) IGF-I. A previous 16-week pilot-study included six...... HIV-infected patients on stable HAART to receive rhGH 0.7 mg/day, which increased total (+117%, P

  17. Screening a strain of Aspergillus niger and optimization of fermentation conditions for degradation of aflatoxin B₁.

    Science.gov (United States)

    Zhang, Wei; Xue, Beibei; Li, Mengmeng; Mu, Yang; Chen, Zhihui; Li, Jianping; Shan, Anshan

    2014-11-13

    Aflatoxin B₁, a type of highly toxic mycotoxin produced by some species belonging to the Aspergillus genus, such as Aspergillus flavus and Aspergillus parasiticus, is widely distributed in feed matrices. Here, coumarin was used as the sole carbon source to screen microorganism strains that were isolated from types of feed ingredients. Only one isolate (ND-1) was able to degrade aflatoxin B₁ after screening. ND-1 isolate, identified as a strain of Aspergillus niger using phylogenetic analysis on the basis of 18S rDNA, could remove 26.3% of aflatoxin B₁ after 48 h of fermentation in nutrient broth (NB). Optimization of fermentation conditions for aflatoxin B₁ degradation by selected Aspergillus niger was also performed. These results showed that 58.2% of aflatoxin B₁ was degraded after 24 h of culture under the optimal fermentation conditions. The aflatoxin B₁ degradation activity of Aspergillus niger supernatant was significantly stronger than cells and cell extracts. Furthermore, effects of temperature, heat treatment, pH, and metal ions on aflatoxin B₁ degradation by the supernatant were examined. Results indicated that aflatoxin B₁ degradation of Aspergillus niger is enzymatic and this process occurs in the extracellular environment.

  18. Intelligent and fine regulation of microbial cell factory based on riboswitches%核糖核酸开关用于微生物细胞工厂的智能与精细调控

    Institute of Scientific and Technical Information of China (English)

    赵雨佳; 张根林; 周晓宏; 李春

    2015-01-01

    Construction of the microbial cell factory is one of the developmental directions of current green chemical industry. The microbial cell factory is a kind of recombined microorganism and its metabolic and regulatory pathways have been reconstructed by metabolic engineering and synthetic biology to synthetic new compounds or to improve the yield of target production. The microbial metabolic pathway is regulated by two points: environment and genetic information. The cell maintains its homeostasis by global transcription factors, messenger molecules and feedback inhibition when the circumstance is changed. Meanwhile, the cell is affected by its own genetic circulate through transcription, translation and post-translational modification to regulate the expression of target gene. The riboswitches are RNA elements which change their conformation when bind to specific ligands such as ions, sugar derivatives, amino acids, nucleic acid derivatives and coenzymes to regulate the process of transcription, translation and splicing of mRNA. The riboswitches are natural biosensors and bioeffectors which can be designed as the intelligent molecular tools to fine regulate microbial cell factories. Using riboswitches in the microbial cell factory can extend the application in the field of chemical, pharmaceutical, environmental protection and food production.%利用代谢工程与合成生物技术对细胞内复杂的代谢网络和调控网络进行重构和改造,以建立合成新化合物或提高目标产物产量的微生物细胞工厂是当今绿色化工技术发展的方向之一。微生物代谢途径的调控受环境和遗传的双重影响,细胞通过全局转录因子、信使分子和反馈抑制等方式响应环境变化来维持细胞的内稳态;同时细胞还受自身遗传基因线路的调控,在转录、翻译以及翻译后修饰过程中调控特定基因的表达。核糖核酸开关是一类调控基因线路表达的 RNA 元件,通过与

  19. Treatment of both native and deamidated gluten peptides with an endo-peptidase from Aspergillus niger prevents stimulation of gut-derived gluten-reactive T cells from either children or adults with celiac disease.

    Science.gov (United States)

    Toft-Hansen, Henrik; Rasmussen, Karina S; Staal, Anne; Roggen, Erwin L; Sollid, Ludvig M; Lillevang, Søren T; Barington, Torben; Husby, Steffen

    2014-08-01

    Celiac disease (CD) is characterized by an inappropriate immunological reaction against gluten driven by gluten-specific CD4+ T cells. We screened 25 proteases and tested 10 for their potential to degrade gluten in vitro. Five proteases were further tested for their ability to prevent the proliferative response by a gluten-specific CD4+ T cell clone and seven gluten-reactive T cell lines to protease-digested gluten peptides. A proline-specific endo-peptidase from Aspergillus niger (AnP2) was particularly efficient at diminishing proliferation after stimulation with cleaved antigen, and could completely block the response against both native and deamidated gluten peptides. We found that AnP2 was efficient down to a 1:64 protease:substrate ratio (w:w). When AnP2 was tested in assays using seven gluten-reactive T cell lines from individual CD patients (three adults and four children), the response to gluten was diminished in all cases. Our study indicates a therapeutic benefit of AnP2 to CD patients.

  20. Selection arena in Aspergillus nidulans

    NARCIS (Netherlands)

    Bruggeman, J.; Debets, A.J.M.; Hoekstra, R.F.

    2004-01-01

    The selection arena hypothesis states that overproduction of zygotes-a widespread phenomenon in animals and plants-can be explained as a mechanism of progeny choice. As a similar mechanism, the ascomycetous fungus Aspergillus nidulans may overproduce dikaryotic fruit initials, hereafter called dikar

  1. Aspergillus mediastinitis after cardiac surgery

    Directory of Open Access Journals (Sweden)

    Marie-Josée Caballero

    2016-03-01

    Conclusion: The clinical features of postoperative Aspergillus mediastinitis may be paucisymptomatic, emphasizing the need for a low index of suspicion in cases of culture-negative mediastinitis or in indolent wound infections. In addition to surgical debridement, the central component of antifungal therapy should include amphotericin B or voriconazole.

  2. Long-term insulin-like growth factor-I expression in skeletal muscles attenuates the enhanced in vitro proliferation ability of the resident satellite cells in transgenic mice

    Science.gov (United States)

    Chakravarthy, M. V.; Fiorotto, M. L.; Schwartz, R. J.; Booth, F. W.

    2001-01-01

    Insulin-like growth factor-I (IGF-I) overexpression for 1-month in mouse skeletal muscle increases satellite cell proliferation potential. However, it is unknown whether this beneficial enhancement by IGF-I expression would persist over a longer-term duration in aged mice. This is an important issue to address if a prolonged course of IGF-I is to be used clinically in muscle-wasting conditions where satellite cells may become limiting. Using the IGF-I transgenic (IGF-I Tg) mouse that selectively expresses the IGF-I transgene in striated muscles, we found that 18-months of continuous IGF-I overexpression led to a loss in the enhanced in vitro proliferative capacity of satellite cells from Tg skeletal muscles. Also 18-month-old IGF-I Tg satellite cells lost the enhanced BrdU incorporation, greater pRb and Akt phosphorylations, and decreased p27(Kip1) levels initially observed in cells from 1-month-old IGF-I Tg mice. The levels of those biochemical markers reverted to similar values seen in the 18-months WT littermates. These findings, therefore, suggest that there is no further beneficial effect on enhancing satellite cell proliferation ability with persistent long-term expression of IGF-I in skeletal muscles of these transgenic mice.

  3. Multiple Segment Factorial Vignette Designs

    Science.gov (United States)

    Ganong, Lawrence H.; Coleman, Marilyn

    2006-01-01

    The multiple segment factorial vignette design (MSFV) combines elements of experimental designs and probability sampling with the inductive, exploratory approach of qualitative research. MSFVs allow researchers to investigate topics that may be hard to study because of ethical or logistical concerns. Participants are presented with short stories…

  4. Identification of thermostable β-xylosidase activities produced by Aspergillus brasiliensis and Aspergillus niger

    DEFF Research Database (Denmark)

    Pedersen, Mads; Lauritzen, Henrik Klitgaard; Frisvad, Jens Christian

    2007-01-01

    Twenty Aspergillus strains were evaluated for production of extracellular cellulolytic and xylanolytic activities. Aspergillus brasiliensis, A. niger and A. japonicus produced the highest xylanase activities with the A. brasiliensis and A. niger strains producing thermostable beta...

  5. Identification of thermostable beta-xylosidase activities produced by Aspergillus brasiliensis and Aspergillus niger

    DEFF Research Database (Denmark)

    Pedersen, Mads; Lauritzen, H.K.; Frisvad, Jens Christian

    2007-01-01

    Twenty Aspergillus strains were evaluated for production of extracellular cellulolytic and xylanolytic activities. Aspergillus brasiliensis, A. niger and A. japonicus produced the highest xylanase activities with the A. brasiliensis and A. niger strains producing thermostable beta...

  6. Involvement of the opportunistic pathogen Aspergillus tubingensis in osteomyelitis of the maxillary bone : a case report

    NARCIS (Netherlands)

    Bathoorn, Erik; Salazar, Natalia Escobar; Sepehrkhouy, Shahrzad; Meijer, Martin; de Cock, Hans; Haas, Pieter-Jan

    2013-01-01

    Background: Aspergillus tubingensis is a black Aspergillus belonging to the Aspergillus section Nigri, which includes species that morphologically resemble Aspergillus niger. Recent developments in species determination have resulted in clinical isolates presumed to be Aspergillus niger being reclas

  7. Albumin Enhances Caspofungin Activity against Aspergillus Species by Facilitating Drug Delivery to Germinating Hyphae.

    Science.gov (United States)

    Ioannou, Petros; Andrianaki, Aggeliki; Akoumianaki, Tonia; Kyrmizi, Irene; Albert, Nathaniel; Perlin, David; Samonis, George; Kontoyiannis, Dimitrios P; Chamilos, Georgios

    2015-12-07

    The modest in vitro activity of echinocandins against Aspergillus implies that host-related factors augment the action of these antifungal agents in vivo. We found that, in contrast to the other antifungal agents (voriconazole, amphotericin B) tested, caspofungin exhibited a profound increase in activity against various Aspergillus species under conditions of cell culture growth, as evidenced by a ≥4-fold decrease in minimum effective concentrations (MECs) (P = 0. 0005). Importantly, the enhanced activity of caspofungin against Aspergillus spp. under cell culture conditions was strictly dependent on serum albumin and was not observed with the other two echinocandins, micafungin and anidulafungin. Of interest, fluorescently labeled albumin bound preferentially on the surface of germinating Aspergillus hyphae, and this interaction was further enhanced upon treatment with caspofungin. In addition, supplementation of cell culture medium with albumin resulted in a significant, 5-fold increase in association of fluorescently labeled caspofungin with Aspergillus hyphae (P caspofungin, with albumin acting as a potential carrier molecule to facilitate antifungal drug delivery to Aspergillus hyphae.

  8. Pulmonary infections by the fungus aspergillus

    Directory of Open Access Journals (Sweden)

    Rao P

    1979-01-01

    Full Text Available Five cases of respiratory infection by Aspergillus fumigates are described. Species of aspergillus is ubiquitous in nature. Therefore, repeated demonstration of fungus, serological evidence tend radiological findings are essential for diagnosis. Potassium iodide is a useful drug in aspergillus infection of the lung when other drugs are not available. Injection Emetine hydrochloride is promising as a therapeutic agent in pulmonary aspergillosis, where the lung parenchyma is involved.

  9. Unilateral cutaneous emboli of Aspergillus.

    Science.gov (United States)

    Watsky, K L; Eisen, R N; Bolognia, J L

    1990-09-01

    A 40-year-old white woman with acute nonlymphocytic leukemia, which relapsed despite bone marrow transplantation and various chemotherapeutic regimens, developed fever and neutropenia. Her fever was unresponsive to broad-spectrum antibiotics, and on hospital day 53 she developed purpuric macules with necrotic centers on her left hand and forearm. Frozen sections of lesional skin were stained with Grocott's methenamine-silver and showed hyphae consistent with a species of Aspergillus; culture of the skin biopsy specimen yielded a pure culture of Aspergillus flavus. Localization of the emboli to the left upper extremity was subsequently explained by magnetic resonance imaging scan of the chest demonstrating invasion of the left subclavian artery by a pulmonary aspergilloma.

  10. Photon Factory Activity Report, 1994

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1995-12-31

    Photon Factory Activity Report no.12 deals with our activities in the period from October 1993 through September 1994. We operate two light sources at the Photon Factory; the 2.5-GeV Photon Factory storage ring, which is a dedicated light source, and the 6.5-GeV TRISTAN Accumulation Ring, which is parasitically used as a light source. We keep more than seventy experimental stations at two facilities, and accept experiments primarily according to approval by the Program Advisory Committee. The number of proposals to the Photon Factory has been still growing. Three-hundred eighty two proposals were approved by the PAC in FY1994, which is an increase by thirteen percent compared to the previous year. Remarkable was growth in biology proposals, particularly proposals in protein crystallography. In FY 1994, we accepted approximately 20,000 man-days as general users, and almost ten percent of them were from abroad. We always open the facility to users, not only domestic but also international. Recently we have been concentrating our effort to upgrading of the light sources and reconstruction of the experimental stations to keep the Photon Factory an attractive research facility in the forthcoming years. We have already started a program of reducing the emittance of the 2.5-GeV storage ring, which now operates with an emittance of 110 nm-rad, to 27 nm-rad by modifying the lattice, with the goal of operation at the reduced emittance in the fall of 1997. We also have conceived of a conversion of the TRISTAN Accumulation Ring to a dedicated light source of high energies. The on-going TRISTAN project will terminate by the end of 1995, and the TRISTAN Main Ring will be converted to a new B-Factory. At this moment, the TRISTAN Accumulation Ring will be disused as the injector to the Main Ring, and conversion of the AR to a dedicated light source becomes possible. (J.P.N.)

  11. Atypical Aspergillus parasiticus isolates from pistachio with aflR gene nucleotide insertion identical to Aspergillus sojae

    Science.gov (United States)

    Aflatoxins are the most toxic and carcinogenic secondary metabolites produced primarily by the filamentous fungi Aspergillus flavus and Aspergillus parasiticus. The toxins cause devastating economic losses because of strict regulations on distribution of contaminated products. Aspergillus sojae are...

  12. 27 CFR 40.69 - Factory premises.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 2 2010-04-01 2010-04-01 false Factory premises. 40.69... PROCESSED TOBACCO Qualification Requirements for Manufacturers of Tobacco Products § 40.69 Factory premises. The premises to be used by a manufacturer of tobacco products as his factory may consist of more...

  13. Iatrogenic aspergillus infection of the central nervous system in a pregnant woman

    Directory of Open Access Journals (Sweden)

    Lokuhetty Menaka

    2009-07-01

    Full Text Available A healthy postnatal woman succumbed to fulminant iatrogenic Aspergillus infection of the central nervous system, following accidental inoculation into the subarachnoid space at spinal anesthesia, during an outbreak of Aspergillus meningitis in Sri Lanka. Autopsy revealed extensive Aspergillus meningitis and culture confirmed Aspergillus fumigatus. The thalamic parenchyma in the brain was invaded by fungal hyphae producing necrotizing angitis with thrombosis, thalamic infarcts and fungal abscesses. The directional growth of fungal hyphae from the extra-luminal side of blood vessels towards the lumen favored extension from the brain parenchyma over hematogenous spread. The spinal parenchyma was resistant to fungal invasion in spite of the heavy growth within the spinal meninges and initial inoculation at spinal level. Modulation of the immune response in pregnancy with depression of selective aspects of cell-mediated immunity probably contributed to rapid spread within the subarachnoid space, to involve the brain parenchyma leading to clinical deterioration and death.

  14. Aspergillus section Nidulantes (formerly Emericella): Polyphasic taxonomy, chemistry and biology

    DEFF Research Database (Denmark)

    Chen, A.J.; Frisvad, Jens Christian; Sun, B.D.

    2016-01-01

    Aspergillus section Nidulantes includes species with striking morphological characters, such as biseriate conidiophores with brown-pigmented stipes, and if present, the production of ascomata embedded in masses of Hülle cells with often reddish brown ascospores. The majority of species in this se......Aspergillus section Nidulantes includes species with striking morphological characters, such as biseriate conidiophores with brown-pigmented stipes, and if present, the production of ascomata embedded in masses of Hülle cells with often reddish brown ascospores. The majority of species...... in this section have a sexual state, which were named Emericella in the dual name nomenclature system. In the present study, strains belonging to subgenus Nidulantes were subjected to multilocus molecular phylogenetic analyses using internal transcribed spacer region (ITS), partial β-tubulin (BenA), calmodulin...

  15. RIKEN RI Beam Factory project

    Energy Technology Data Exchange (ETDEWEB)

    Yano, Yasushige; Goto, Akira; Katayama, Takeshi [Institute of Physical and Chemical Research, Wako, Saitama (Japan)

    1997-03-01

    The RARF proposes `RIKEN RI Beam Factory` as a next facility-expanding project. The factory makes it the primary aim to provide RI (Radioactive Isotope) beams covering over the whole atomic-mass range with the world-highest intensity in a wide energy range up to several hundreds MeV/nucleon. These RI beams are generated by the fragmentation of high-intensity heavy-ion beams. For the efficient production heavy-ion energies will be boosted up to over 100 MeV/nucleon even for very heavy ions by a K2500-MeV superconducting ring cyclotron serving as a post accelerator of the existing K540-MeV ring cyclotron. A new type of experimental installation called `MUSES` (Multi-USe Experimental Storage rings) will be constructed as well. With MUSES, various types of unique colliding experiments will become possible. (author)

  16. Exotic Hadrons from B Factories

    Science.gov (United States)

    Fulsom, Bryan

    2017-01-01

    The first generation of B-Factories, BaBar and Belle, operated over the previous decade and produced many world-leading measurements related to flavor physics. One of the most important discoveries was that of an apparent four-quark particle, named X(3872). It was the first of a growing X, Y, Z alphabet of exotic hadrons, now numbering more than a dozen, found by the e + e - collider experiments. These multi-quark states represent an unusual departure from the standard description that hadronic matter consists of only two or three quarks. These discoveries have led to the emergence of a new category of physics within heavy meson spectroscopy. This talk will review some of these key experimental results, and highlight the potential of the next generation B-Factory, Belle II, as it begins operation in the coming year.

  17. Handbook factory planning and design

    CERN Document Server

    Wiendahl, Hans-Peter; Nyhuis, Peter

    2015-01-01

    This handbook introduces a methodical approach and pragmatic concept for the planning and design of changeable factories that act in strategic alliances to supply the ever-changing needs of the global market. In the first part, the change drivers of manufacturing enterprises and the resulting new challenges are considered in detail with focus on an appropriate change potential. The second part concerns the design of the production facilities and systems on the factory levels work place, section, building and site under functional, organisational, architectural and strategic aspects keeping in mind the environmental, health and safety aspects including corporate social responsibility. The third part is dedicated to the planning and design method that is based on a synergetic interaction of process and space. The accompanying project management of the planning and construction phase and the facility management for the effective utilization of the built premises close the book. -        Concise overview o...

  18. Modulation of antimicrobial metabolites production by the fungus Aspergillus parasiticus

    Directory of Open Access Journals (Sweden)

    Adriana A.P. Bracarense

    2014-01-01

    Full Text Available Biosynthesis of active secondary metabolites by fungi occurs as a specific response to the different growing environments. Changes in this environment alter the chemical and biological profiles leading to metabolites diversification and consequently to novel pharmacological applications. In this work, it was studied the influence of three parameters (fermentation length, medium composition and aeration in the biosyntheses of antimicrobial metabolites by the fungus Aspergillus parasiticus in 10 distinct fermentation periods. Metabolism modulation in two culturing media, CYA and YES was evaluated by a 2² full factorial planning (ANOVA and on a 2³ factorial planning, role of aeration, medium composition and carbohydrate concentration were also evaluated. In overall, 120 different extracts were prepared, their HPLC profiles were obtained and the antimicrobial activity against A. flavus, C. albicans, E. coli and S. aureus of all extracts was evaluated by microdilution bioassay. Yield of kojic acid, a fine chemical produced by the fungus A. parasiticus was determined in all extracts. Statistical analyses pointed thirteen conditions able to modulate the production of bioactive metabolites by A. parasiticus. Effect of carbon source in metabolites diversification was significant as shown by the changes in the HPLC profiles of the extracts. Most of the extracts presented inhibition rates higher than that of kojic acid as for the extract obtained after 6 days of fermentation in YES medium under stirring. Kojic acid was not the only metabolite responsible for the activity since some highly active extracts showed to possess low amounts of this compound, as determined by HPLC.

  19. Effect of gamma radiation on Aspergillus flavus and Aspergillus ochraceus ultrastructure and mycotoxin production

    Energy Technology Data Exchange (ETDEWEB)

    Ribeiro, J. [Departamento de Microbiologia e Inmunologia Veterinaria, Universidad Federal Rural de Rio de Janeiro (UFRRJ) (Brazil); Cavaglieri, L., E-mail: lcavaglieri@arnet.com.a [Departamento de Microbiologia e Inmunologia, Universidad Nacional de Rio Cuarto (UNRC), Rio Cuarto, Cordoba (Argentina); Member of Consejo Nacional de Investigaciones Cientificas y Tecnologicas (CIC-CONICET) (Argentina); Vital, H. [Centro Tecnologico do Exercito (CTEx), Secao de Defesa Nuclear, Rio de Janeiro (Brazil); Cristofolini, A.; Merkis, C. [Departamento de Microscopia Electronica, Universidad Nacional de Rio Cuarto. Ruta 36 km 601 (5800) Rio Cuarto (Argentina); Astoreca, A. [Departamento de Microbiologia e Inmunologia, Universidad Nacional de Rio Cuarto (UNRC), Rio Cuarto, Cordoba (Argentina); Member of Consejo Nacional de Investigaciones Cientificas y Tecnologicas (CIC-CONICET) (Argentina); Orlando, J.; Caru, M. [Departamento de Ciencias Ecologicas, Facultad de Ciencias, Universidad de Chile, Santiago (Chile); Dalcero, A. [Departamento de Microbiologia e Inmunologia, Universidad Nacional de Rio Cuarto (UNRC), Rio Cuarto, Cordoba (Argentina); Member of Consejo Nacional de Investigaciones Cientificas y Tecnologicas (CIC-CONICET) (Argentina); Rosa, C.A.R. [Departamento de Microbiologia e Inmunologia Veterinaria, Universidad Federal Rural de Rio de Janeiro (UFRRJ) (Brazil); Member of Consejo Nacional de Pesquisas (CNPq) (Brazil)

    2011-05-15

    The aim of this work was to study the effect of gamma radiation (2 kGy) on Aspergillus flavus and Aspergillus ochraceus ultrastructure. Moreover, the influence on aflatoxin B{sub 1} and ochratoxin A production was also observed. Irradiated A. flavus strain showed a dull orangish colony while control strain showed the typical green color. Minor differences were observed on stipes, metulae and conidia size between control and irradiated A. flavus and A. ochraceus strains. Irradiated fungi showed ultrastructural changes on cell wall, plasmalema and cytoplasm levels. The levels of mycotoxins produced by irradiated strains were two times greater than those produced by control strains. Successive transferences of irradiated strains on malt extract agar allowed the fungus to recuperate morphological characteristics. Although minor changes in the fungal morphology were observed, ultrastructural changes at cell wall level and the increase of mycotoxin production ability were observed. Inappropriate storage of irradiated food and feed would allow the development of potentially more toxicogenic fungal propagules.

  20. Results from the B Factories

    Energy Technology Data Exchange (ETDEWEB)

    Bevan, A.; /Queen Mary, U. of London

    2009-01-08

    These proceedings are based on lectures given at the Helmholtz International Summer School Heavy Quark Physics at the Bogoliubov Laboratory of Theoretical Physics, Dubna, Russia, during August 2008. I review the current status of CP violation in B meson decays from the B factories. These results can be used, along with measurements of the sides of the Unitarity Triangle, to test the CKM mechanism. In addition I discuss experimental studies of B decays to final states with 'spin-one' particles.

  1. Photon Factory activity report, 1995

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1996-12-31

    The Photon Factory at the National Laboratory for High Energy Physics is a national facility for scientific research utilizing synchrotron radiation. Although the Photon Factory operator the linear injector, two light sources including the 2.5 GeV storage ring and the 6.5 GeV TRISTAN accumulation Ring as well as a major fraction of their beamlines and experimental station. This report is covered the period from October 1994 to September 1995. The total number of proposals by this PAC was 399 in 1995. Facility development projects currently in progress include the following, TRISTAN Super Light Facility (TSLF) project, VUV-FEL project, KEKB project and Slow-positron Source. This report contents outline of the Photon Factory, introduction, scientific disciplines, electronic properties of condensed matters, atomic and molecular science, X-ray imaging, radiobiology using synchrotron radiation, structural properties of condensed matters, structural properties of solid surfaces and adsorbates, structure and function of proteins, theoretical researches, experimental facilities, beamlines, new instrumentation, AR Upgrade, collaborations, projects, user`s short reports, list of published papers 1994/95. (S.Y.)

  2. 带状疱疹疫苗生产中2BS细胞细胞工厂培养工艺的建立%Development of culture procedure for 2BS cell factory during production of herpes zoster vaccine

    Institute of Scientific and Technical Information of China (English)

    梁雪; 刘延威; 张宇; 石磊; 杨晓叶; 李生军; 沈红杰

    2014-01-01

    Objective To develop a culture procedure for 2BS cell factory during production of herpes zoster vaccine.Methods The 2BS cells were resuscitated (passage 28) and subcultured to passages 35 and 36,then to the ten layer cell factory at mass ratios of 1 ∶ 4 and 1 ∶ 2 respectively.The cells on each layer were added with various volumes (200,300 and 400 ml) of medium,observed for morphology by microscopy and counted.The cell factory was inoculated with varicellazoster virus at a MOI of 0.008 and prepared into live attenuated vaccine.The bulk and final product of vaccine were determined for titers.The final product was subjected to stability test at 37 ℃.The residual antibiotics and bovine albumin contents were determined by ELISA,while the endotoxin content by gel method,and the moisture content by volume titration method.Results Dense 2BS cell monolayer with high quality was prepared by subculture at a mass ratio of 1 ∶ 2 and addition of 300 ml of medium,of which the CPE reached more than 80% at time of harvest.The cells with CPE were round,most of which were in typical shape.The titers of bulk and final product of vaccine prepared with cells subcultured at a mass ratio of 1 ∶ 4 were significantly lower than those at 1 ∶ 2.However,the titer in stability test at 37 ℃ of final product of vaccine prepared with the cells subcultured at a mass ratio of 1 ∶ 4 failed to met the requirements of not less than 4.6 lgPFU/ml after storage for one week.Other indexes met the relevant requirements.Conclusion A culture procedure for 2BS cell factory during production of herpes zoster vaccine was developed,by which dense 2BS cells with even and high quality were prepared.%目的 建立带状疱疹疫苗生产中2BS细胞在细胞工厂中的培养工艺.方法 将2BS细胞复苏(28代)后,连续传代至35、36代,再分别按1∶4、1∶2比例传至10层细胞工厂中,采用不同的加液量(200、300、400 ml/层)进行培养,显微镜下观察细胞形态,并

  3. The role of thiol species in the hypertolerance of Aspergillus sp. P37 to arsenic.

    Science.gov (United States)

    Cánovas, David; Vooijs, Riet; Schat, Henk; de Lorenzo, Víctor

    2004-12-01

    Aspergillus sp. P37 is an arsenate-hypertolerant fungus isolated from a river in Spain with a long history of contamination with metals. This strain is able to grow in the presence of 0.2 M arsenate, i.e. 20-fold higher than the reference strain, Aspergillus nidulans TS1. Although Aspergillus sp. P37 reduces As(V) to As(III), which is slowly pumped out of the cell, the measured efflux of oxyanions is insufficient to explain the high tolerance levels of this strain. To gain an insight into this paradox, the accumulation of acid-soluble thiol species in Aspergillus sp. P37 when exposed to arsenic was compared with that of the arsenic-sensitive A. nidulans TS1 strain. Increasing levels of arsenic in the medium did not diminish the intracellular pool of reduced glutathione in Aspergillus sp. P37, in sharp contrast with the decline of glutathione in A. nidulans under the same conditions. Furthermore, concentrations of arsenic that were inhibitory for the sensitive A. nidulans strain (e.g. 50 mM and above) provoked a massive formation of vacuoles filled with thiol species. Because the major fraction of the cellular arsenic was present as the glutathione conjugate As(GS)3, it is plausible that the arsenic-hypertolerant phenotype of Aspergillus sp. P37 is in part due to an enhanced capacity to maintain a large intracellular glutathione pool under conditions of arsenic exposure and to sequester As(GS)3 in vacuoles. High pressure liquid chromatography analysis of cell extracts revealed that the contact of Aspergillus sp. P37 (but not A. nidulans) with high arsenic concentrations (> or =150 mM) induced the production of small quantities of a distinct thiol species indistinguishable from plant phytochelatin-2. Yet, we argue that phytochelatins do not explain arsenic resistance in Aspergillus, and we advocate the role of As(GS)3 complexes in arsenic detoxification.

  4. Enhanced succinic acid production in Aspergillus saccharolyticus by heterologous expression of fumarate reductase from Trypanosoma brucei.

    Science.gov (United States)

    Yang, Lei; Lübeck, Mette; Ahring, Birgitte K; Lübeck, Peter S

    2016-02-01

    Aspergillus saccharolyticus exhibits great potential as a cell factory for industrial production of dicarboxylic acids. In the analysis of the organic acid profile, A. saccharolyticus was cultivated in an acid production medium using two different pH conditions. The specific activities of the enzymes, pyruvate carboxylase (PYC), malate dehydrogenase (MDH), and fumarase (FUM), involved in the reductive tricarboxylic acid (rTCA) branch, were examined and compared in cells harvested from the acid production medium and a complete medium. The results showed that ambient pH had a significant impact on the pattern and the amount of organic acids produced by A. saccharolyticus. The wild-type strain produced higher amount of malic acid and succinic acid in the pH buffered condition (pH 6.5) compared with the pH non-buffered condition. The enzyme assays showed that the rTCA branch was active in the acid production medium as well as the complete medium, but the measured enzyme activities were different depending on the media. Furthermore, a soluble NADH-dependent fumarate reductase gene (frd) from Trypanosoma brucei was inserted and expressed in A. saccharolyticus. The expression of the frd gene led to an enhanced production of succinic acid in frd transformants compared with the wild-type in both pH buffered and pH non-buffered conditions with highest amount produced in the pH buffered condition (16.2 ± 0.5 g/L). This study demonstrates the feasibility of increasing succinic acid production through the cytosolic reductive pathway by genetic engineering in A. saccharolyticus.

  5. Ecophysiological characterization of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger isolated from grapes in Spanish vineyards.

    Science.gov (United States)

    García-Cela, E; Crespo-Sempere, A; Ramos, A J; Sanchis, V; Marin, S

    2014-03-03

    The aim of this study was to evaluate the diversity of black aspergilli isolated from berries from different agroclimatic regions of Spain. Growth characterization (in terms of temperature and water activity requirements) of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger was carried out on synthetic grape medium. A. tubingensis and A. niger showed higher maximum temperatures for growth (>45 °C versus 40-42 °C), and lower minimum aw requirements (0.83 aw versus 0.87 aw) than A. carbonarius. No differences in growth boundaries due to their geographical origin were found within A. niger aggregate isolates. Conversely, A. carbonarius isolates from the hotter and drier region grew and produced OTA at lower aw than other isolates. However, little genetic diversity in A. carbonarius was observed for the microsatellites tested and the same sequence of β-tubulin gene was observed; therefore intraspecific variability did not correlate with the geographical origin of the isolates or with their ability to produce OTA. Climatic change prediction points to drier and hotter climatic scenarios where A. tubingensis and A. niger could be even more prevalent over A. carbonarius, since they are better adapted to extreme high temperature and drier conditions.

  6. Insulin-like growth factor-I receptor (IGF-IR) targeting with monoclonal antibody cixutumumab (IMC-A12) inhibits IGF-I action in endometrial cancer cells.

    Science.gov (United States)

    Attias-Geva, Zohar; Bentov, Itay; Ludwig, Dale L; Fishman, Ami; Bruchim, Ilan; Werner, Haim

    2011-07-01

    Specific insulin-like growth factor-I receptor (IGF-IR) targeting emerged in recent years as a promising therapeutic strategy in cancer. Endometrial cancer is the most common gynaecological cancer in the Western world. The aim of this study was to evaluate the potential of cixutumumab (IMC-A12, ImClone Systems), a fully human monoclonal antibody against the IGF-IR, to inhibit IGF-I-mediated biological actions and cell signalling events in four endometrial carcinoma-derived cell lines (ECC-1, Ishikawa, USPC-1 and USPC-2). Our results demonstrate that cixutumumab was able to block the IGF-I-induced autophosphorylation of the IGF-IR. In addition, the PI3K and MAPK downstream signalling pathways were also inactivated by cixutumumab in part of the cell lines. Prolonged (24h and 48h) exposures to cixutumumab reduced IGF-IR expression. Furthermore, confocal microscopy of GFP-tagged receptors shows that cixutumumab treatment led to IGF-IR redistribution from the cell membrane to the cytoplasm. Antiapoptotic effects were evaluated by cleavage of caspase 3 and PARP, and mitogenicity and transformation by proliferation and cell cycle assays. Results obtained showed that cixutumumab abrogated the IGF-I-stimulated increase in proliferation rate, and increased caspase-3 and PARP cleavage, two markers of apoptosis. Of importance, cixutumumab had no effect neither on insulin receptor (IR) expression nor on IGF-I activation of IR. In summary, in a cellular model of endometrial cancer cixutumumab was able to inhibit the IGF-I-induced activation of intracellular cascades, apoptosis and proliferation.

  7. The preliminary study on the effects of growth hormone and insulin-like growth factor-I on κ-casein synthesis in bovine mammary epithelial cells in vitro.

    Science.gov (United States)

    Wang, M Z; Ji, Y; Wang, C; Chen, L M; Wang, H R; Loor, J J

    2016-04-01

    The effects of growth hormone (GH) and insulin-like growth factor-I (IGF-I) on protein synthesis and gene expression of κ-casein in bovine mammary epithelial cell in vitro were studied. The treatments were designed as follows: the growth medium without serum was set as the control group, while the treatments were medium supplemented with GH (100 ng/ml), IGF-I (100 ng/ml), and GH (100 ng/ml) + IGF-I (100 ng/ml). The quantity of κ-casein protein was measured by ELISA, and the κ-casein gene (CSN3) expression was examined by real-time quantitative PCR (RT-qPCR). Compared with the control group, all the experimental groups had greater (p  0.05). Furthermore, no synergistic effect of GH and IGF-I was observed for both the κ-casein concentration and CSN3 expression. It is therefore concluded that GH or IGF-I can independently promote the expression of CSN3 in bovine mammary epithelial cells in vitro.

  8. Infectious keratitis caused by Aspergillus tubingensis

    NARCIS (Netherlands)

    Kredics, L.; Varga, J.; Kocsube, S.; Rajaraman, R.; Raghavan, A.; Doczi, I.; Bhaskar, M.; Nemeth, T.M.; Antal, Z.; Venkatapathy, N.; Vagvolgyi, C.; Samson, R.A.; Chockaiya, M.; Palanisamy, M.

    2009-01-01

    PURPOSE: To report 2 cases of keratomycosis caused by Aspergillus tubingensis. METHODS: The therapeutic courses were recorded for 2 male patients, 52 and 78 years old, with fungal keratitis caused by black Aspergillus strains. Morphological examination of the isolates was carried out on malt extract

  9. Sporulation inhibited secretion in Aspergillus niger

    NARCIS (Netherlands)

    Krijgsheld, P.

    2013-01-01

    Aspergillus niger is abundantly found in nature. It degrades dead material of plants and animals but can also be a pathogen of these organisms. Aspergillus niger is also important for mankind because it is one of the main organisms used for the industrial production of enzymes. These enzymes are rel

  10. Microbially induced and microbially catalysed precipitation: two different carbonate factories

    Science.gov (United States)

    Meister, Patrick

    2016-04-01

    The landmark paper by Schlager (2003) has revealed three types of benthic carbonate production referred to as "carbonate factories", operative at different locations at different times in Earth history. The tropical or T-factory comprises the classical platforms and fringing reefs and is dominated by carbonate precipitation by autotrophic calcifying metazoans ("biotically controlled" precipitation). The cool or C-factory is also biotically controlled but via heterotrophic, calcifying metazoans in cold and deep waters at the continental margins. A further type is the mud-mound or M-factory, where carbonate precipitation is supported by microorganisms but not controlled by a specific enzymatic pathway ("biotically induced" precipitation). How exactly the microbes influence precipitation is still poorly understood. Based on recent experimental and field studies, the microbial influence on modern mud mound and microbialite growth includes two fundamentally different processes: (1) Metabolic activity of microbes may increase the saturation state with respect to a particular mineral phase, thereby indirectly driving the precipitation of the mineral phase: microbially induced precipitation. (2) In a situation, where a solution is already supersaturated but precipitation of the mineral is inhibited by a kinetic barrier, microbes may act as a catalyser, i.e. they lower the kinetic barrier: microbially catalysed precipitation. Such a catalytic effect can occur e.g. via secreted polymeric substances or specific chemical groups on the cell surface, at which the minerals nucleate or which facilitate mechanistically the bonding of new ions to the mineral surface. Based on these latest developments in microbialite formation, I propose to extend the scheme of benthic carbonate factories of Schlager et al. (2003) by introducing an additional branch distinguishing microbially induced from microbially catalysed precipitation. Although both mechanisms could be operative in a M-factory

  11. New taxa in Aspergillus section Usti

    DEFF Research Database (Denmark)

    Samson, R. A.; Varga, J.; Meijer, M.

    2011-01-01

    Based on phylogenetic analysis of sequence data, Aspergillus section Usti includes 21 species, inducing two teleomorphic species Aspergillus heterothallicus (=Emericella heterothallica) and Fennellia monodii. Aspergillus germanicus sp. nov. was isolated from indoor air in Germany. This species has...... identical ITS sequences with A. insuetus CBS 119.27, but is clearly distinct from that species based on beta-tubulin and calmodulin sequence data. This species is unable to grow at 37 degrees C, similarly to A. keveii and A. insuetus. Aspergillus carlsbadensis sp. nov. was isolated from the Carlsbad Caverns...... National Park in New Mexico. This taxon is related to, but distinct from a dade including A. calidoustus, A. pseudodeflectus, A. insuetus and A. keveii on all trees. This species is also unable to grow at 37 degrees C, and acid production was not observed on CREA. Aspergillus californicus sp. nov...

  12. Vaccination approaches against opportunistic fungal infections caused by Aspergillus fumigatus.

    Science.gov (United States)

    Reichard, Utz; Herrmann, Sahra; Asif, Abdul R

    2014-01-01

    Although innate immunity primarily combats systemic infections of opportunistic fungi such as Aspergillus and Candida spp., acquired and protective immunoreactions were observed long ago in animal trials following sublethal systemic infections caused by viable fungi or after challenging animals with inactivated fungal cells. Based on these observations, fungal antigens should exist which mediate such protective immunoreactions and have in part already been identified. In this context, this review focuses primarily on the various approaches that have been used to identify protection-mediating Aspergillus-antigens and their rationale. Emphasis is placed on screening methods that have exploited genetic or proteomic approaches on the basis of the corresponding fungal genome projects. Thereby, a survey and description is given of the antigens so far known to be capable of inducing immune responses that protect animals against acquiring lethal systemic aspergillosis.

  13. Muon Colliders and Neutrino Factories

    Energy Technology Data Exchange (ETDEWEB)

    Kaplan, Daniel M. [IIT, Chicago

    2015-05-29

    Muon colliders and neutrino factories are attractive options for future facilities aimed at achieving the highest lepton-antilepton collision energies and precision measurements of Higgs boson and neutrino mixing matrix parameters. The facility performance and cost depend on how well a beam of muons can be cooled. Recent progress in muon cooling design studies and prototype tests nourishes the hope that such facilities could be built starting in the coming decade. The status of the key technologies and their various demonstration experiments is summarized. Prospects "post-P5" are also discussed.

  14. Muon Colliders and Neutrino Factories

    CERN Document Server

    Kaplan, Daniel M

    2014-01-01

    Muon colliders and neutrino factories are attractive options for future facilities aimed at achieving the highest lepton-antilepton collision energies and precision measurements of Higgs boson and neutrino mixing matrix parameters. The facility performance and cost depend on how well a beam of muons can be cooled. Recent progress in muon cooling design studies and prototype tests nourishes the hope that such facilities could be built starting in the coming decade. The status of the key technologies and their various demonstration experiments is summarized. Prospects "post-P5" are also discussed.

  15. Aspergillus Osteomyelitis of the Skull.

    Science.gov (United States)

    Nicholson, Simon; King, Richard; Chumas, Paul; Russell, John; Liddington, Mark

    2016-07-01

    Osteomyelitis of the craniofacial skeleton is rare, with fungal pathogens least commonly implicated. The authors present 2 patients of osteomyelitis of the skull caused by Aspergillus spp. and discuss the diagnosis, clinicopathological course, and management strategies.Late recurrence seen in this type of infection warrants long-term follow-up and a high index of suspicion for the clinical signs associated with recurrence.Such patients would benefit from their surgical debridement being planned and managed via a specialist craniofacial unit, so as to utilize the most aesthetically sensitive approach and the experience of specialists from several surgical disciplines.

  16. Glucoamylase I of Black Aspergillus

    OpenAIRE

    Medda, Sukumar; Saha, Badal Chandra; Ueda, Seinosuke

    1982-01-01

    Glucoamylase I of black Aspergillus was purified by (NH_4)_2SO_4 precipitation, ethanol fractionation, ion exchange chromatography on DEAE-cellulose, preparative isoelectric focusing and Sephadex G-100 gel filtration. The enzyme thus purified was found to contain no α-amylase and appeared to be homogeneous in polyacrylamide gel electrophoresis. The isoelectric point of glucoamylase I was at pH 3.4. The optimum conditions for its action on boiled soluble starch were at 60℃ and pH 4.5. The enzy...

  17. Two novel species of Aspergillus section Nigri from indoor air

    Science.gov (United States)

    Aspergillus collinsii, Aspergillus floridensis, and Aspergillus trinidadensis are described as novel uniseriate species of Aspergillus section Nigri isolated from air samples. To describe the species we used phenotypes from 7-d Czapek yeast extract agar culture (CYA) and malt extract agar culture (M...

  18. Ultrastructural Characterization of Zika Virus Replication Factories

    Directory of Open Access Journals (Sweden)

    Mirko Cortese

    2017-02-01

    Full Text Available A global concern has emerged with the pandemic spread of Zika virus (ZIKV infections that can cause severe neurological symptoms in adults and newborns. ZIKV is a positive-strand RNA virus replicating in virus-induced membranous replication factories (RFs. Here we used various imaging techniques to investigate the ultrastructural details of ZIKV RFs and their relationship with host cell organelles. Analyses of human hepatic cells and neural progenitor cells infected with ZIKV revealed endoplasmic reticulum (ER membrane invaginations containing pore-like openings toward the cytosol, reminiscent to RFs in Dengue virus-infected cells. Both the MR766 African strain and the H/PF/2013 Asian strain, the latter linked to neurological diseases, induce RFs of similar architecture. Importantly, ZIKV infection causes a drastic reorganization of microtubules and intermediate filaments forming cage-like structures surrounding the viral RF. Consistently, ZIKV replication is suppressed by cytoskeleton-targeting drugs. Thus, ZIKV RFs are tightly linked to rearrangements of the host cell cytoskeleton.

  19. 1,25(OH)2D3 and VDR Signaling Pathways Regulate the Inhibition of Dectin-1 Caused by Cyclosporine A in Response to Aspergillus Fumigatus in Human Corneal Epithelial Cells

    Science.gov (United States)

    Xia, Yiping; Zhao, Guiqiu; Lin, Jing; Li, Cui; Cong, Lin; Jiang, Nan; Xu, Qiang; Wang, Qian

    2016-01-01

    Background The objective of this study is to observe whether cyclosporine A (CsA) inhibits the expression of dectin-1 in human corneal epithelial cells infected with Aspergillus fumigatus (A. fumigatus) and to investigate the molecular mechanisms of the inhibition. Methods Immortalized human corneal epithelial cells (HCECs) were pretreated with 1,25(OH)2D3 and VDR inhibitor for 1 h, and then they were pretreated with CsA for 12h. After these pretreatments, the HCECs were stimulated with A. fumigatus and curdlan respectively, and the expression of dectin-1 and proinflammatory cytokines (IL-1β and TNF-α) were detected by RT-PCR, western blot and ELISA. Results Dectin-1 mRNA and dectin-1 protein expression increased when HCECs were stimulated with A. fumigatus or curdlan, and CsA inhibited the dectin-1 expression both in mRNA and protein levels specifically. Dectin-1 and proinflammatory cytokine expression levels were higher when HCECs were pretreated with VDR inhibitor and CsA compared to pretreatment with CsA alone, while dectin-1 and proinflammatory cytokine levels were lower when HCECs were pretreated with 1,25(OH)2D3 and CsA compared to pretreatment with CsA alone. Conclusions These data provide evidence that CsA can inhibit the expression of dectin-1 and proinflammatory cytokines through dectin-1 when HCECs are stimulated by A. fumigatus or curdlan. The active form of vitamin D, 1,25(OH)2D3, and VDR signaling pathway regulate the inhibition of CsA. The inhibition is enhanced by 1,25(OH)2D3, and the VDR inhibitor suppresses the inhibition. PMID:27755569

  20. Screening a Strain of Aspergillus niger and Optimization of Fermentation Conditions for Degradation of Aflatoxin B1

    Directory of Open Access Journals (Sweden)

    Wei Zhang

    2014-11-01

    Full Text Available Aflatoxin B1, a type of highly toxic mycotoxin produced by some species belonging to the Aspergillus genus, such as Aspergillus flavus and Aspergillus parasiticus, is widely distributed in feed matrices. Here, coumarin was used as the sole carbon source to screen microorganism strains that were isolated from types of feed ingredients. Only one isolate (ND-1 was able to degrade aflatoxin B1 after screening. ND-1 isolate, identified as a strain of Aspergillus niger using phylogenetic analysis on the basis of 18S rDNA, could remove 26.3% of aflatoxin B1 after 48 h of fermentation in nutrient broth (NB. Optimization of fermentation conditions for aflatoxin B1 degradation by selected Aspergillus niger was also performed. These results showed that 58.2% of aflatoxin B1 was degraded after 24 h of culture under the optimal fermentation conditions. The aflatoxin B1 degradation activity of Aspergillus niger supernatant was significantly stronger than cells and cell extracts. Furthermore, effects of temperature, heat treatment, pH, and metal ions on aflatoxin B1 degradation by the supernatant were examined. Results indicated that aflatoxin B1 degradation of Aspergillus niger is enzymatic and this process occurs in the extracellular environment.

  1. Aspergillus niger contains the cryptic phylogenetic species A. awamori

    DEFF Research Database (Denmark)

    Perrone, Giancarlo; Stea, Gaetano; Epifani, Filomena

    2011-01-01

    Aspergillus section Nigri is an important group of species for food and medical mycology, and biotechnology. The Aspergillus niger ‘aggregate’ represents its most complicated taxonomic subgroup containing eight morphologically indistinguishable taxa: A. niger, Aspergillus tubingensis, Aspergillus...... acidus, Aspergillus brasiliensis, Aspergillus costaricaensis, Aspergillus lacticoffeatus, Aspergillus piperis, and Aspergillus vadensis. Aspergillus awamori, first described by Nakazawa, has been compared taxonomically with other black aspergilli and recently it has been treated as a synonym of A. niger....... Phylogenetic analyses of sequences generated from portions of three genes coding for the proteins β-tubulin (benA), calmodulin (CaM), and the translation elongation factor-1 alpha (TEF-1α) of a population of A. niger strains isolated from grapes in Europe revealed the presence of a cryptic phylogenetic species...

  2. Ribonuclease Production by Aspergillus species

    Directory of Open Access Journals (Sweden)

    Gomes Eleni

    1998-01-01

    Full Text Available Ribonuclease production by Aspergillus flavipes, A. sulphureus and A. fischeri in semi-synthetic medium, after 24-144 hours at 30ºC under shaking, was studied. After cultivation, the medium was separated from micelia by filtration and the resultant solution was used as enzymatic extract. The highest amount of biomass and RNase was obtained after 96 hours of cultivation. The enzymes produced by three species presented similar characteristics, with optimum temperature at 55ºC and two peaks of activity at pH 4.5 and 7.0. A. flavipes RNases were more sensitive to temperature: 50% of the initial activity was lost after 1 hour at 70ºC. After this heat treatment, RNase of A. sulphureus lost 30% of this activity and that of A. fischeri only 16%. The nucleotides released by enzimatic hydrolysis of RNA were separated by ion exchange chromatography in a AG-1X8-formiate column and identified by paper chromatography. This procedure indicated that the raw enzymatic extract of Aspergillus flavipes is able to hydrolyze RNA, releasing 3'-nucleotides monophosphate at pH 4.5 and 3' and 5'-nucleotides monophosphate at pH 7.0 and 8.5. This result suggests that this strain produces two different types of RNase, one acidic and other alcaline, with different specificities.

  3. Inhibition of secretion by asexual reproduction in Aspergillus niger

    NARCIS (Netherlands)

    Wang, Fengfeng

    2014-01-01

    The aim of this Thesis was to study mechanism involved in zonal secretion in colonies of A. niger with emphasis on sporulation inhibited secretion. These mechanisms may be targets to improve A. nigeras a cell factory. I focused in particular on A. niger homologues of fluG and veA that are known spor

  4. Secondary metabolite profiles and antifungal drug susceptibility of Aspergillus fumigatus and closely related species, Aspergillus lentulus, Aspergillus udagawae, and Aspergillus viridinutans.

    Science.gov (United States)

    Tamiya, Hiroyuki; Ochiai, Eri; Kikuchi, Kazuyo; Yahiro, Maki; Toyotome, Takahito; Watanabe, Akira; Yaguchi, Takashi; Kamei, Katsuhiko

    2015-05-01

    The incidence of Aspergillus infection has been increasing in the past few years. Also, new Aspergillus fumigatus-related species, namely Aspergillus lentulus, Aspergillus udagawae, and Aspergillus viridinutans, were shown to infect humans. These fungi exhibit marked morphological similarities to A. fumigatus, albeit with different clinical courses and antifungal drug susceptibilities. The present study used liquid chromatography/time-of-flight mass spectrometry to identify the secondary metabolites secreted as virulence factors by these Aspergillus species and compared their antifungal susceptibility. The metabolite profiles varied widely among A. fumigatus, A. lentulus, A. udagawae, and A. viridinutans, producing 27, 13, 8, and 11 substances, respectively. Among the mycotoxins, fumifungin, fumiquinazoline A/B and D, fumitremorgin B, gliotoxin, sphingofungins, pseurotins, and verruculogen were only found in A. fumigatus, whereas auranthine was only found in A. lentulus. The amount of gliotoxin, one of the most abundant mycotoxins in A. fumigatus, was negligible in these related species. In addition, they had decreased susceptibility to antifungal agents such as itraconazole and voriconazole, even though metabolites that were shared in the isolates showing higher minimum inhibitory concentrations than epidemiological cutoff values were not detected. These strikingly different secondary metabolite profiles may lead to the development of more discriminative identification protocols for such closely related Aspergillus species as well as improved treatment outcomes.

  5. Aspergillus uvarum sp. nov., an uniseriate black Aspergillus species isolated from grapes in Europe

    DEFF Research Database (Denmark)

    Perrone, Giancarlo; Varga, János; Susca, Antonia

    2008-01-01

    A novel species, Aspergillus uvarum sp. nov., is described within Aspergillus section Nigri. This species can be distinguished from other black aspergilli based on internal transcribed spacers (ITS), beta-tubulin and calmodulin gene sequences, by AFLP analysis and by extrolite profiles. Aspergillus...... uvarum sp. nov. isolates produced secalonic acid, common to other Aspergillus japonicus-related taxa, and geodin, erdin and dihydrogeodin, which are not produced by any other black aspergilli. None of the isolates were found to produce ochratoxin A. The novel species is most closely related to two...... atypical strains of Aspergillus aculeatus, CBS 114.80 and CBS 620.78, and was isolated from grape berries in Portugal, Italy, France, Israel, Greece and Spain. The type strain of Aspergillus uvarum sp. nov. is IMI 388523(T)=CBS 127591(T)= ITEM 4834(T)= IBT26606(T)....

  6. Genetic diversity of Aspergillus species isolated from onychomycosis and Aspergillus hongkongensis sp. nov., with implications to antifungal susceptibility testing.

    Science.gov (United States)

    Tsang, Chi-Ching; Hui, Teresa W S; Lee, Kim-Chung; Chen, Jonathan H K; Ngan, Antonio H Y; Tam, Emily W T; Chan, Jasper F W; Wu, Andrea L; Cheung, Mei; Tse, Brian P H; Wu, Alan K L; Lai, Christopher K C; Tsang, Dominic N C; Que, Tak-Lun; Lam, Ching-Wan; Yuen, Kwok-Yung; Lau, Susanna K P; Woo, Patrick C Y

    2016-02-01

    Thirteen Aspergillus isolates recovered from nails of 13 patients (fingernails, n=2; toenails, n=11) with onychomycosis were characterized. Twelve strains were identified by multilocus sequencing as Aspergillus spp. (Aspergillus sydowii [n=4], Aspergillus welwitschiae [n=3], Aspergillus terreus [n=2], Aspergillus flavus [n=1], Aspergillus tubingensis [n=1], and Aspergillus unguis [n=1]). Isolates of A. terreus, A. flavus, and A. unguis were also identifiable by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The 13th isolate (HKU49(T)) possessed unique morphological characteristics different from other Aspergillus spp. Molecular characterization also unambiguously showed that HKU49(T) was distinct from other Aspergillus spp. We propose the novel species Aspergillus hongkongensis to describe this previously unknown fungus. Antifungal susceptibility testing showed most Aspergillus isolates had low MICs against itraconazole and voriconazole, but all Aspergillus isolates had high MICs against fluconazole. A diverse spectrum of Aspergillus species is associated with onychomycosis. Itraconazole and voriconazole are probably better drug options for Aspergillus onychomycosis.

  7. FleA Expression in Aspergillus fumigatus Is Recognized by Fucosylated Structures on Mucins and Macrophages to Prevent Lung Infection.

    Science.gov (United States)

    Kerr, Sheena C; Fischer, Gregory J; Sinha, Meenal; McCabe, Orla; Palmer, Jonathan M; Choera, Tsokyi; Lim, Fang Yun; Wimmerova, Michaela; Carrington, Stephen D; Yuan, Shaopeng; Lowell, Clifford A; Oscarson, Stefan; Keller, Nancy P; Fahy, John V

    2016-04-01

    The immune mechanisms that recognize inhaled Aspergillus fumigatus conidia to promote their elimination from the lungs are incompletely understood. FleA is a lectin expressed by Aspergillus fumigatus that has twelve binding sites for fucosylated structures that are abundant in the glycan coats of multiple plant and animal proteins. The role of FleA is unknown: it could bind fucose in decomposed plant matter to allow Aspergillus fumigatus to thrive in soil, or it may be a virulence factor that binds fucose in lung glycoproteins to cause Aspergillus fumigatus pneumonia. Our studies show that FleA protein and Aspergillus fumigatus conidia bind avidly to purified lung mucin glycoproteins in a fucose-dependent manner. In addition, FleA binds strongly to macrophage cell surface proteins, and macrophages bind and phagocytose fleA-deficient (∆fleA) conidia much less efficiently than wild type (WT) conidia. Furthermore, a potent fucopyranoside glycomimetic inhibitor of FleA inhibits binding and phagocytosis of WT conidia by macrophages, confirming the specific role of fucose binding in macrophage recognition of WT conidia. Finally, mice infected with ΔfleA conidia had more severe pneumonia and invasive aspergillosis than mice infected with WT conidia. These findings demonstrate that FleA is not a virulence factor for Aspergillus fumigatus. Instead, host recognition of FleA is a critical step in mechanisms of mucin binding, mucociliary clearance, and macrophage killing that prevent Aspergillus fumigatus pneumonia.

  8. Bioproduction, characterization, anticancer and antioxidant activities of extracellular melanin pigment produced by newly isolated microbial cell factories Streptomyces glaucescens NEAE-H

    Science.gov (United States)

    El-Naggar, Noura El-Ahmady; El-Ewasy, Sara M.

    2017-01-01

    In this present study, a newly isolated strain, Streptomyces sp. NEAE-H, capable of producing high amount of black extracellular melanin pigment on peptone-yeast extract iron agar and identified as Streptomyces glaucescens NEAE-H. Plackett–Burman statistical design was conducted for initial screening of 17 independent (assigned) variables for their significances on melanin pigment production by Streptomyces glaucescens NEAE-H. The most significant factors affecting melanin production are incubation period, protease-peptone and ferric ammonium citrate. The levels of these significant variables and their interaction effects were optimized by using face-centered central composite design. The maximum melanin production (31.650 μg/0.1 ml) and tyrosinase activity (6089.10 U/ml) were achieved in the central point runs under the conditions of incubation period (6 days), protease-peptone (5 g/L) and ferric ammonium citrate (0.5 g/L). Melanin pigment was recovered by acid-treatment. Higher absorption of the purified melanin pigment was observed in the UV region at 250 nm. It appeared to have defined small spheres by scanning electron microscopy imaging. The maximum melanin yield was 350 mg dry wt/L of production medium. In vitro anticancer activity of melanin pigment was assayed against skin cancer cell line using MTT assay. The IC50 value was 16.34 ± 1.31 μg/ml for melanin and 8.8 ± 0.5 μg/ml for standard 5-fluorouracil. PMID:28195138

  9. N-cadherin as a receptor for adhesion and endocytosis of Aspergillus fumigatus by human umbilical vein endothelial cells%N-钙黏蛋白在烟曲霉黏附及侵袭内皮细胞中的作用

    Institute of Scientific and Technical Information of China (English)

    徐小勇; 施毅; 张鹏鹏; 申玉英; 张峰; 宋勇

    2010-01-01

    Objective To study the receptor for adhesion and endocytosis of Aspergillus fumigatus hyphae by human umbilical vein endothelial cells (HUVEC). Methods Aspergillus fumigatus hyphae were incubated with the total protein of HUVEC for investigating the binding of N-cadherin and the fungus. After the model of adhesion and endocytosis of Aspergillus fumigatus by HUVEC was established, the capacity of adhesion and endocytosis was evaluated with the presence of the antibody to N-cadherin. Results Ncadherin sticked to the surface of Aspergillus fumigatus. Adhesion and endocytosis were inhibited with the presence of the antibody to N-cadherin. Conclusion N-cadherin is a receptor for adhesion and endocytosis of Aspergillus fumigatus by HUVEC.%目的 探讨N-钙黏蛋白在内皮细胞黏附吞噬烟曲霉过程中的作用.方法 观察提取人脐静脉内皮细胞蛋白与烟曲霉的结合过程,了解N-钙黏蛋白是否可与烟曲霉结合,建立内皮细胞黏附及吞噬烟曲霉的体外模型,通过单克隆抗体阻断上皮细胞膜受体N-钙黏蛋白,再次观察脐静脉内皮细胞黏附及吞噬烟曲霉情况.结果 脐静脉内皮细胞膜蛋白N-钙黏蛋白可与烟曲霉结合,抗体阻断N-钙黏蛋白后,脐静脉内皮细胞黏附和吞噬烟曲霉能力明显下降.结论 N-钙黏蛋白是脐静脉内皮细胞黏附吞噬烟曲霉孢子的相关受体.

  10. Constitutive expression of fluorescent protein by Aspergillus var. niger and Aspergillus carbonarius to monitor fungal colonization in maize plants.

    Science.gov (United States)

    Palencia, Edwin Rene; Glenn, Anthony Elbie; Hinton, Dorothy Mae; Bacon, Charles Wilson

    2013-09-01

    Aspergillus niger and Aspergillus carbonarius are two species in the Aspergillus section Nigri (black-spored aspergilli) frequently associated with peanut (Arachis hypogea), maize (Zea mays), and other plants as pathogens. These infections are symptomless and as such are major concerns since some black aspergilli produce important mycotoxins, ochratoxins A, and the fumonisins. To facilitate the study of the black aspergilli-maize interactions with maize during the early stages of infections, we developed a method that used the enhanced yellow fluorescent protein (eYFP) and the monomeric red fluorescent protein (mRFP1) to transform A. niger and A. carbonarius, respectively. The results were constitutive expressions of the fluorescent genes that were stable in the cytoplasms of hyphae and conidia under natural environmental conditions. The hyphal in planta distribution in 21-day-old seedlings of maize were similar wild type and transformants of A. niger and A. carbonarius. The in planta studies indicated that both wild type and transformants internally colonized leaf, stem and root tissues of maize seedlings, without any visible disease symptoms. Yellow and red fluorescent strains were capable of invading epidermal cells of maize roots intercellularly within the first 3 days after inoculation, but intracellular hyphal growth was more evident after 7 days of inoculation. We also tested the capacity of fluorescent transformants to produce ochratoxin A and the results with A. carbonarius showed that this transgenic strain produced similar concentrations of this secondary metabolite. This is the first report on the in planta expression of fluorescent proteins that should be useful to study the internal plant colonization patterns of two ochratoxigenic species in the Aspergillus section Nigri.

  11. Gentamicin removal in submerged fermentation using the novel fungal strain Aspergillus terreus FZC3

    Science.gov (United States)

    Liu, Yuanwang; Chang, Huiqing; Li, Zhaojun; Zhang, Cheng; Feng, Yao; Cheng, Dengmiao

    2016-10-01

    Social concern and awareness of the potential risk posed by environmental residues of antibiotics such as gentamicin in the development of antibiotic resistance genes have increased. The present study used laboratory-scale experiments to develop methods for gentamicin removal from the environment. A fungus, strain FZC3, which could remove gentamicin in submerged fermentation, was isolated from solid waste and sewage water from a gentamicin production factory. The fungus was identified as Aspergillus terreus by sequencing the PCR-amplified ITS fragments of its rRNA-coding genes and by its morphology. The gentamicin removal efficiency exceeded 95% by day 7 under optimized culture conditions. The results showed that both biosorption and biodegradation were involved. We speculated that Aspergillus terreus FZC3 absorbed gentamicin and subsequently degraded it. We also found that Aspergillus terreus FZC3 survived and maintained a high bioremediation efficiency over a wide pH range, indicating its potential for future use in the large-scale bioremediation of gentamicin.

  12. Metabolites from marine fungus Aspergillus sp.

    Digital Repository Service at National Institute of Oceanography (India)

    Wahidullah, S.; Rajmanickam, R.; DeSouza, L.

    Chemical examination of a methanolic extract of the marine fungus, Aspergillus sp., isolated from marine grass environment, yielded a steroid, ergosterol peroxide (1), and a mixture of known glyceride esters (2,3) of unsaturated fatty acids...

  13. New species in Aspergillus section Terrei

    DEFF Research Database (Denmark)

    Samson, R. A.; Peterson, S. W.; Frisvad, Jens Christian

    2011-01-01

    Section Terrei of Aspergillus was studied using a polyphasic approach including sequence analysis of parts of the beta-tubulin and calmodulin genes and the ITS region, macro- and micromorphological analyses and examination of extrolite profiles to describe three new species in this section. Based...... on phylogenetic analysis of calmodulin and beta-tubulin sequences seven lineages were observed among isolates that have previously been treated as A. terreus and its subspecies by Raper & Fennell (1965) and others. Aspergillus alabamensis, A. terreus var. floccosus, A. terreus var. africanus, A. terreus var....... floccosus, A. terreus var. africanus, A. terreus var. aureus, while Aspergillus hortai is recognised at species level. Aspergillus terreus NRRL 4017 is described as the new species A. pseudoterreus. Also included in section Terrei are some species formerly placed in sections Flavipedes and Versicolores. A...

  14. Phylogeny and subgeneric taxonomy of Aspergillus

    DEFF Research Database (Denmark)

    Peterson, S.W.; Varga, Janos; Frisvad, Jens Christian

    2008-01-01

    The phylogeny of the genus Aspergillus and its teleomorphs is discussed based on multilocus sequence data. DNA sequence analysis was used to formulate a nucleotide sequence framework of the genus and to analyze character changes in relationship to the phylogeny hypothesized from the DNA sequence...... analysis. The sequence data used to delimitate subgeneric taxa included partial calmodulin, rDNA and RNA polymerase gene sequences. In our phylogenic structure of Aspergillus extrolite data of the various Aspergillus taxa collected from ex-type cultures and numerous other isolates are also discussed. A new...... subgeneric classification is proposed which includes 8 subgenera and 22 sections within the Aspergillus genus. Characteristics of these taxa are shortly discussed in this chapter....

  15. Lipase production by recombinant strains of Aspergillus niger expressing a lipase-encoding gene from Thermomyces lanuginosus

    DEFF Research Database (Denmark)

    Prathumpai, Wai; Flitter, S.J.; Mcintyre, Mhairi

    2004-01-01

    Two recombinant strains of Aspergillus niger (NW 297-14 and NW297-24) producing a heterologous lipase from Thermomyces lanuginosus were constructed. The heterologous lipase was expressed using the TAKA amylase promoter from Aspergillus oryzae. The production kinetics of the two strains on different...... shows that it is possible to obtain high productivities of heterologous fungal enzymes in A. niger. However, SDS-PAGE analysis showed that most of the produced lipase was bound to the cell wall....

  16. A Tau-Charm Factory at CEBAF

    Energy Technology Data Exchange (ETDEWEB)

    Seth, K.K. [Northwestern Univ., Evanston, IL (United States)

    1994-04-01

    It is proposed that a Tau Charm Factory represents a natural extension of CEBAF into higher energy domains. The exciting nature of the physics of charm quarks and tau leptons is briefly reviewed and it is suggested that the concept of a linac-ring collider as a Tau Charm Factory at CEBAF should be seriously studied.

  17. Three new species of Aspergillus section Flavi isolated from almonds and maize in Portugal

    Science.gov (United States)

    Three new aflatoxin-producing species belonging to Aspergillus section Flavi are described, Aspergillus mottae, Aspergillus sergii and Aspergillus transmontanensis. These species were isolated from Portuguese almonds and maize. An investigation examining morphology, extrolites and molecular data was...

  18. 27 CFR 40.114 - Extension or curtailment of factory.

    Science.gov (United States)

    2010-04-01

    ... of factory. 40.114 Section 40.114 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND... Products Changes in Location of Factory § 40.114 Extension or curtailment of factory. Where a tobacco products factory is to be changed to an extent which will make inaccurate the description of the factory...

  19. Charged Particle Optics in Circular Higgs Factory

    Energy Technology Data Exchange (ETDEWEB)

    Cai, Yunhai [SLAC National Accelerator Lab., Menlo Park, CA (United States)

    2015-02-26

    Similar to a super B-factory, a circular Higgs factory will require strong focusing systems near the interaction points and a low-emittance lattice in arcs to achieve a factory luminosity. At electron beam energy of 120 GeV, beamstrahlung effects during the collision pose an additional challenge to the collider design. In particular, a large momentum acceptance at 2 percent level is necessary to retain an adequate beam lifetime. This turns out to be the most challenging aspect in the design of circular Higgs factory. In this paper, an example will be provided to illustrate the beam dynamics in circular Higgs factory, emphasizing on the chromatic optics. Basic optical modules and advanced analysis will be presented. Most important, we will show that 2% momentum aperture is achievable

  20. Factorial invariance in multilevel confirmatory factor analysis.

    Science.gov (United States)

    Ryu, Ehri

    2014-02-01

    This paper presents a procedure to test factorial invariance in multilevel confirmatory factor analysis. When the group membership is at level 2, multilevel factorial invariance can be tested by a simple extension of the standard procedure. However level-1 group membership raises problems which cannot be appropriately handled by the standard procedure, because the dependency between members of different level-1 groups is not appropriately taken into account. The procedure presented in this article provides a solution to this problem. This paper also shows Muthén's maximum likelihood (MUML) estimation for testing multilevel factorial invariance across level-1 groups as a viable alternative to maximum likelihood estimation. Testing multilevel factorial invariance across level-2 groups and testing multilevel factorial invariance across level-1 groups are illustrated using empirical examples. SAS macro and Mplus syntax are provided.

  1. Post-genomic insights into the plant polysaccharide degradation potential of Aspergillus nidulans and comparison to Aspergillus niger and Aspergillus oryzae

    DEFF Research Database (Denmark)

    Coutinho, Pedro M.; Andersen, Mikael Rørdam; Kolenova, Katarina

    2009-01-01

    The plant polysaccharide degradative potential of Aspergillus nidulans was analysed in detail and compared to that of Aspergillus niger and Aspergillus oryzae using a combination of bioinformatics, physiology and transcriptomics. Manual verification indicated that 28.4% of the A. nidulans ORFs an...

  2. Modelling Aspergillus flavus growth and aflatoxins production in pistachio nuts.

    Science.gov (United States)

    Marín, Sonia; Ramos, Antonio J; Sanchis, V

    2012-12-01

    Aflatoxins (AFs) are the main contaminants in pistachio nuts. AFs production in pistachio has been attributed to Aspergillus flavus. The aim of this study was to apply existing models to predict growth and AFs production by an A. flavus isolated from pistachios as a function of moisture content and storage temperature of pistachios in order to test their usefulness and complementarities. A full factorial design was used: the moisture content levels assayed were 10, 15, 20, 25 and 30% and incubation temperatures were 10, 15, 20, 25, 30, 37 and 42 °C. Both kinetic and probability models were built to predict growth of the strain under the assayed conditions. Among the assayed models, cardinal ones gave a good quality fit for radial growth rate data. Moreover, the progressive approach, which was developed based on a reduced number of experimental points led to an improved prediction in the validation step. This is quite significant as may allow for improved experimental designs, less costly than full factorial ones. Probability model proved to be concordant in 91% of the calibration set observations. Even though the validation set included conditions around the growth/no-growth interface, there was a 100% agreement in the predictions from the data set (n = 16, cut off = 0.5) after 60 days. Similarly, the probability for AF presence was rightly predicted in 89% of the cases. According to our results EC maximum aflatoxin levels would be surpassed in a period as short as 1 month if pistachio nuts reach 20 °C, unless %mc is ≤10%.

  3. Characterization of specific immune responses to different Aspergillus antigens during the course of invasive Aspergillosis in hematologic patients.

    Science.gov (United States)

    Potenza, Leonardo; Vallerini, Daniela; Barozzi, Patrizia; Riva, Giovanni; Forghieri, Fabio; Beauvais, Anne; Beau, Remi; Candoni, Anna; Maertens, Johan; Rossi, Giulio; Morselli, Monica; Zanetti, Eleonora; Quadrelli, Chiara; Codeluppi, Mauro; Guaraldi, Giovanni; Pagano, Livio; Caira, Morena; Del Giovane, Cinzia; Maccaferri, Monica; Stefani, Alessandro; Morandi, Uliano; Tazzioli, Giovanni; Girardis, Massimo; Delia, Mario; Specchia, Giorgina; Longo, Giuseppe; Marasca, Roberto; Narni, Franco; Merli, Francesco; Imovilli, Annalisa; Apolone, Giovanni; Carvalho, Agostinho; Comoli, Patrizia; Romani, Luigina; Latgè, Jean Paul; Luppi, Mario

    2013-01-01

    Several studies in mouse model of invasive aspergillosis (IA) and in healthy donors have shown that different Aspergillus antigens may stimulate different adaptive immune responses. However, the occurrence of Aspergillus-specific T cells have not yet been reported in patients with the disease. In patients with IA, we have investigated during the infection: a) whether and how specific T-cell responses to different Aspergillus antigens occur and develop; b) which antigens elicit the highest frequencies of protective immune responses and, c) whether such protective T cells could be expanded ex-vivo. Forty hematologic patients have been studied, including 22 patients with IA and 18 controls. Specific T cells producing IL-10, IFN-γ, IL-4 and IL-17A have been characterized through enzyme linked immunospot and cytokine secretion assays on 88 peripheral blood (PB) samples, by using the following recombinant antigens: GEL1p, CRF1p, PEP1p, SOD1p, α1-3glucan, β1-3glucan, galactomannan. Specific T cells were expanded through short term culture. Aspergillus-specific T cells producing non-protective interleukin-10 (IL-10) and protective interferon-gamma (IFN-γ) have been detected to all the antigens only in IA patients. Lower numbers of specific T cells producing IL-4 and IL-17A have also been shown. Protective T cells targeted predominantly Aspergillus cell wall antigens, tended to increase during the IA course and to be associated with a better clinical outcome. Aspergillus-specific T cells could be successfully generated from the PB of 8 out of 8 patients with IA and included cytotoxic subsets able to lyse Aspergillus hyphae. Aspergillus specific T-cell responses contribute to the clearance of the pathogen in immunosuppressed patients with IA and Aspergillus cell wall antigens are those mainly targeted by protective immune responses. Cytotoxic specific T cells can be expanded from immunosuppressed patients even during the infection by using the above mentioned antigens

  4. Effects of dietary supplementation of Aspergillus originated prebiotic (Fermacto on performance and small intestinal morphology of broiler chickens fed diluted diets

    Directory of Open Access Journals (Sweden)

    Zarbakht Ansari Pirsaraei

    2010-02-01

    Full Text Available Fermacto, that it is commercially a fermentation product of Aspergillus orizae, is proven to increase digestion efficiency of the gut. In this experiment, 450 one day old Cobb 500 broiler chicks (mixed-sex were used in a completely randomized design with 3×2 factorial arrangement. There were three levels of Fermacto (0, 0.15 and 0.3% for 42 days and two levels of diet dilution using rice hulls (0 and 40% at 10-21 days of age. The treatments consisted of 3 replicates with 25 chicks per each. Administration of Fermacto did not influence feed intake or feed conversion ratio, but the daily weight gain improved. Diluted diets significantly increased body weights and decreased feed intake; however, this manipulation didn’t affect feed conversion ratios. There was no interaction between prebiotic supplementation and experimental diets dilution for performance traits. Diet dilution significantly decreased abdominal fat pad, villi height and increased epithelial thickness and goblet cell numbers. Dietary supplementation using Fermacto significantly decreased the abdominal fat pad and goblet cell numbers but increased the small intestinal villus heights. There were some conflicting observations for different segments of the small intestine. The 0.3% Fermacto inclusion in experimental diets caused more beneficial effects. Contrary to previous reports on more efficiency of Fermacto on low protein diets, this trend is not seen for the whole diluted diets.

  5. Heterologous expression of the Aspergillus nidulans alcR-alcA system in Aspergillus niger

    NARCIS (Netherlands)

    Nikolaev, I.; Mathieu, M.; Vondervoort, van de P.J.I.; Visser, J.; Felenbok, B.

    2002-01-01

    The inducible and strongly expressed alcA gene encoding alcohol dehydrogenase I from Aspergillus nidulans was transferred together with the activator gene alcR, in the industrial fungus Aspergillus niger. This latter organism does not possess an inducible alc system but has an endogenously constitut

  6. The structure and expression of the pyruvatekinase gene of Aspergillus nidulans and Aspergillus niger.

    NARCIS (Netherlands)

    Graaff, de L.H.

    1989-01-01

    Dit proefschrift "Het pyruvaat kinase gen en zijn expressie in Aspergillus nidulans en Aspergillus niger " beschrijft het onderzoek naar de structuur en expressie van het pyruvaat kinase gen in deze schimmels.De doelstelling van dit onderzoek was de opheldering van de genstructuur van het pyruvaat k

  7. Aspergillus tubingensis and Aspergillus niger as the dominant black Aspergillus, use of simple PCR-RFLP for preliminary differentiation.

    Science.gov (United States)

    Mirhendi, H; Zarei, F; Motamedi, M; Nouripour-Sisakht, S

    2016-03-01

    This work aimed to identify the species distribution of common clinical and environmental isolates of black Aspergilli based on simple restriction fragment length polymorphism (RFLP) analysis of the β-tubulin gene. A total of 149 clinical and environmental strains of black Aspergilli were collected and subjected to preliminary morphological examination. Total genomic DNAs were extracted, and PCR was performed to amplify part of the β-tubulin gene. At first, 52 randomly selected samples were species-delineated by sequence analysis. In order to distinguish the most common species, PCR amplicons of 117 black Aspergillus strains were identified by simple PCR-RFLP analysis using the enzyme TasI. Among 52 sequenced isolates, 28 were Aspergillus tubingensis, 21 Aspergillus niger, and the three remaining isolates included Aspergillus uvarum, Aspergillus awamori, and Aspergillus acidus. All 100 environmental and 17 BAL samples subjected to TasI-RFLP analysis of the β-tubulin gene, fell into two groups, consisting of about 59% (n=69) A. tubingensis and 41% (n=48) A. niger. Therefore, the method successfully and rapidly distinguished A. tubingensis and A. niger as the most common species among the clinical and environmental isolates. Although tardy, the Ehrlich test was also able to differentiate A. tubingensis and A. niger according to the yellow color reaction specific to A. niger. A. tubingensis and A. niger are the most common black Aspergillus in both clinical and environmental isolates in Iran. PCR-RFLP using TasI digestion of β-tubulin DNA enables rapid screening for these common species.

  8. Yeast cell factories on the horizon

    DEFF Research Database (Denmark)

    Nielsen, Jens

    2015-01-01

    For thousands of years, yeast has been used for making beer, bread, and wine. In modern times, it has become a commercial workhorse for producing fuels, chemicals, and pharmaceuticals such as insulin, human serum albumin, and vaccines against hepatitis virus and human papillomavirus. Yeast has al...

  9. Neutrino factories: realization and physics potential

    Energy Technology Data Exchange (ETDEWEB)

    Geer, S.; /Fermilab; Zisman, M.S.; /LBL, Berkeley

    2006-12-01

    Neutrino Factories offer an exciting option for the long-term neutrino physics program. This new type of neutrino facility will provide beams with unique properties. Low systematic uncertainties at a Neutrino Factory, together with a unique and precisely known neutrino flavor content, will enable neutrino oscillation measurements to be made with unprecedented sensitivity and precision. Over recent years, the resulting neutrino factory physics potential has been discussed extensively in the literature. In addition, over the last six years the R&D necessary to realize a Neutrino Factory has been progressing, and has developed into a significant international activity. It is expected that, within about five more years, the initial phase of this R&D program will be complete and, if the community chooses to build this new type of neutrino source within the following decade, neutrino factory technology will be ready for the final R&D phase prior to construction. In this paper (1) an overview is given of the technical ingredients needed for a Neutrino Factory, (2) beam properties are described, (3) the resulting neutrino oscillation physics potential is summarized, (4) a more detailed description is given for one representative Neutrino Factory design, and (5) the ongoing R&D program is summarized, and future plans briefly described.

  10. Changing Charlie and the Chocolate Factory

    OpenAIRE

    2014-01-01

    In this essay the various underlying themes in the two film adaptations of Roald Dahl’s book Charlie and the Chocolate Factory (1964) are examined. These film adaptations, Charlie and the Chocolate Factory (2005) directed by Tim Burton and Willy Wonka and the Chocolate Factory (1971) directed by Mel Stuart are discussed, in order to refute critical views such as found in a review by Bernard Beck. Beck states that Tim Burton’s film is darker and closer to Dahl’s original tale, whilst Mel Stuar...

  11. Crystallization and Preliminary Crystallographic Characterization of Endo-polygalacturonase II from Aspergillus niger

    NARCIS (Netherlands)

    Schröter, K.-H.; Arkema, A.; Kester, H.C.M.; Visser, J.; Dijkstra, B.W.

    1994-01-01

    The endo-polygalacturonase II from Aspergillus niger has been crystallized from an ammonium sulfate solution by the hanging drop method. The crystals belong to the monoclinic space group P2(1), with cell dimensions a = 60.6 Angstrom, b = 152.6 Angstrom, c = 74.0 Angstrom and beta = 91.2 degrees with

  12. Genotoxicity of the cyclo-oxygenase-inhibitor sulindac sulfide in the filamentous fungus Aspergillus nidulans Genotoxicidade de sulfeto de sulindaco em Aspergillus nidulans

    OpenAIRE

    Claudinéia Conationi da Silva Franco; Claudia Tiemi Miyamoto; Juliane Rocha de Sant'Anna; Marialba Avezum Alves Castro-Prado

    2007-01-01

    Sulindac sulfide is a non-steroidal anti-inflammatory drug (NSAID) with chemopreventive effect on human cancer cells. Due to the involvement of the somatic recombination in the carcinogenic process, sulindac sulfide's recombinogenic potential was evaluated by the Homozygotization Index (HI) in the filamentous fungus Aspergillus nidulans. The drug's recombinogenic potential was evaluated by its capacity to induce homozygosis of recessive genes from heterozygous diploid cells. Sulindac sulfide ...

  13. Extrapulmonary Aspergillus infection in patients with CARD9 deficiency

    Science.gov (United States)

    Gazendam, Roel P.; Freeman, Alexandra F.; Hsu, Amy P.; Collar, Amanda L.; Sugui, Janyce A.; Drummond, Rebecca A.; Rongkavilit, Chokechai; Hoffman, Kevin; Henderson, Carolyn; Clark, Lily; Mezger, Markus; Swamydas, Muthulekha; Engeholm, Maik; Schüle, Rebecca; Neumayer, Bettina; Mikelis, Constantinos M.; Pittaluga, Stefania; Prasad, Vinod K.; Singh, Anurag; Milner, Joshua D.; Williams, Kelli W.; Lim, Jean K.; Kwon-Chung, Kyung J.; Holland, Steven M.; Hartl, Dominik; Kuijpers, Taco W.

    2016-01-01

    Invasive pulmonary aspergillosis is a life-threatening mycosis that only affects patients with immunosuppression, chemotherapy-induced neutropenia, transplantation, or congenital immunodeficiency. We studied the clinical, genetic, histological, and immunological features of 2 unrelated patients without known immunodeficiency who developed extrapulmonary invasive aspergillosis at the ages of 8 and 18. One patient died at age 12 with progressive intra-abdominal aspergillosis. The other patient had presented with intra-abdominal candidiasis at age 9, and developed central nervous system aspergillosis at age 18 and intra-abdominal aspergillosis at age 25. Neither patient developed Aspergillus infection of the lungs. One patient had homozygous M1I CARD9 (caspase recruitment domain family member 9) mutation, while the other had homozygous Q295X CARD9 mutation; both patients lacked CARD9 protein expression. The patients had normal monocyte and Th17 cell numbers in peripheral blood, but their mononuclear cells exhibited impaired production of proinflammatory cytokines upon fungus-specific stimulation. Neutrophil phagocytosis, killing, and oxidative burst against Aspergillus fumigatus were intact, but neither patient accumulated neutrophils in infected tissue despite normal neutrophil numbers in peripheral blood. The neutrophil tissue accumulation defect was not caused by defective neutrophil-intrinsic chemotaxis, indicating that production of neutrophil chemoattractants in extrapulmonary tissue is impaired in CARD9 deficiency. Taken together, our results show that CARD9 deficiency is the first known inherited or acquired condition that predisposes to extrapulmonary Aspergillus infection with sparing of the lungs, associated with impaired neutrophil recruitment to the site of infection. PMID:27777981

  14. HiPER Tritium factory elements

    Science.gov (United States)

    Guillaume, Didier

    2011-06-01

    HiPER will include a Tritium target factory. This presentation is an overview. We start from process ideas to go to first sketch passing through safety principles. We will follow the Tritium management process. We need first a gas factory producing the right gas mixture from hydrogen, Deuterium and Tritium storage. Then we could pass through the target factory. It is based on our LMJ single shot experiment and some new development like the injector. Then comes pellet burst and vapour recovery. The Tritium factory has to include the waste recovery, recycling process with gas purification before storage. At least, a nuclear plant is not a classical building. Tritium is also very special... All the design ideas have to be adapted. Many facilities are necessary, some with redundancy. We all have to well known these constraints. Tritium budget will be a major contributor for a material point of view as for a financial one.

  15. Verbal and Nonverbal Communication of Factory Workers

    Science.gov (United States)

    Tway, Patricia

    1976-01-01

    Examines the verbal and nonverbal behavior patterns associated with two speech styles, one formal and the other informal, among factory workers. Available from: Mouton Publishers, Box 482, the Hague, Netherlands. (AM)

  16. Visual evoked potentials in rubber factory workers.

    Science.gov (United States)

    Tandon, O P; Kumar, V

    1997-01-01

    Pattern reversal visual evoked potentials (pVEP) were studied in 39 male rubber factory workers in the age range of 18-55 years and 20 control subjects (aged 18-46 years) not exposed to the rubber factory environment. Results revealed that 20 (51%) rubber factory workers had abnormal latencies of wave P1 (dominant component of pVEP) as per accepted criteria of 99% tolerance limit set for the control group (i.e. any value above mean +3 SD of control was considered abnormal). The section-wise per cent distribution of abnormalities was vulcanization (83%), tubing (75%), calendering (60%), loading (38%) and mixing (14%). This study provides electrophysiological evidence that rubber factory environments affect the conduction processes in optical pathways from their origin in the retina to striate cortex. However, this study has its limitations in not identifying the specific chemical(s) causing these changes in VEP.

  17. Identification of Colour Reconnection using Factorial Correlator

    Institute of Scientific and Technical Information of China (English)

    FU Jing-Hua; LIU Lian-Shou

    2000-01-01

    A new signal is proposed for the colour reconnection in the hadronic decay of W+ W- in e+e- collisions. Using Pythia Monte Carlo it is shown that factorial correlators for W+ and W- without colour reconnection are almost identical to unity, while those for the cases with colour reconnection fall down approximately linearly in the log log plot. This signal, being based on the factorial correlator, is more sensitive than the ones using only averaged quantities.

  18. The physics of the B factories

    CERN Document Server

    Bevan, A J; Mannel, Th; Prell, S; Yabsley, B D; Abe, K; Aihara, H; Anulli, F; Arnaud, N; Aushev, T; Beneke, M; Beringer, J; Bianchi, F; Bigi, I I; Bona, M; Brambilla, N; Brodzicka, J; Chang, P; Charles, M J; Cheng, C H; Cheng, H -Y; Chistov, R; Colangelo, P; Coleman, J P; Drutskoy, A; Druzhinin, V P; Eidelman, S; Eigen, G; Eisner, A M; Faccini, R; Flood, K T; Gambino, P; Gaz, A; Gradl, W; Hayashii, H; Higuchi, T; Hulsbergen, W D; Hurth, T; Iijima, T; Itoh, R; Jackson, P D; Kass, R; Kolomensky, Yu G; Kou, E; Križan, P; Kronfeld, A; Kumano, S; Kwon, Y J; Latham, T E; Leith, D W G S; Lüth, V; Martinez-Vidal, F; Meadows, B T; Mussa, R; Nakao, M; Nishida, S; Ocariz, J; Olsen, S L; Pakhlov, P; Pakhlova, G; Palano, A; Pich, A; Playfer, S; Poluektov, A; Porter, F C; Robertson, S H; Roney, J M; Roodman, A; Sakai, Y; Schwanda, C; Schwartz, A J; Seidl, R; Sekula, S J; Steinhauser, M; Sumisawa, K; Swanson, E S; Tackmann, F; Trabelsi, K; Uehara, S; Uno, S; van der Water, R; Vasseur, G; Verkerke, W; Waldi, R; Wang, M Z; Wilson, F F; Zupan, J; Zupanc, A; Adachi, I; Albert, J; Banerjee, Sw; Ben-Haim, E; Biassoni, P; Cahn, R N; Cartaro, C; Chauveau, J; Chen, C; Chiang, C C; Cowan, R; Dalseno, J; Davier, M; Davies, C; Dingfelder, J C; Echenard, B; Epifanov, D; Fulsom, B G; Gabareen, A M; Gary, J W; Godang, R; Graham, M T; Hafner, A; Hamilton, B; Hartmann, T; Hayasaka, K; Hearty, C; Iwasaki, Y; Khodjamirian, A; Kusaka, A; Kuzmin, A; Lafferty, G D; Lazzaro, A; Li, J; Lindemann, D; Long, O; Lusiani, A; Marchiori, G; Martinelli, M; Miyabayashi, K; Mizuk, R; Mohanty, G B; Muller, D R; Nakazawa, H; Ongmongkolkul, P; Pacetti, S; Palombo, F; Pedlar, T K; Piilonen, L E; Pilloni, A; Poireau, V; Prothmann, K; Pulliam, T; Rama, M; Ratcliff, B N; Roudeau, P; Schrenk, S; Schroeder, T; Schubert, K R; Shen, C P; Shwartz, B; Soffer, A; Solodov, E P; Somov, A; Starič, M; Stracka, S; Telnov, A V; Todyshev, K Yu; Tsuboyama, T; Uglov, T; Vinokurova, A; Walsh, J J; Watanabe, Y; Won, E; Wormser, G; Wright, D H; Ye, S; Zhang, C C; Abachi, S; Abashian, A; Abe, N; Abe, R; Abe, T; Abrams, G S; Adam, I; Adamczyk, K; Adametz, A; Adye, T; Agarwal, A; Ahmed, H; Ahmed, M; Ahmed, S; Ahn, B S; Ahn, H S; Aitchison, I J R; Akai, K; Akar, S; Akatsu, M; Akemoto, M; Akhmetshin, R; Akre, R; Alam, M S; Albert, J N; Aleksan, R; Alexander, J P; Alimonti, G; Allen, M T; Allison, J; Allmendinger, T; Alsmiller, J R G; Altenburg, D; Alwyn, K E; An, Q; Anderson, J; Andreassen, R; Andreotti, D; Andreotti, M; Andress, J C; Angelini, C; Anipko, D; Anjomshoaa, A; Anthony, P L; Antillon, E A; Antonioli, E; Aoki, K; Arguin, J F; A, K; Arisaka, K; Asai, K; Asai, M; Asano, Y; Asgeirsson, D J; Asner, D M; Aso, T; Aspinwall, M L; Aston, D; Atmacan, H; Aubert, B; Aulchenko, V; Ayad, R; Azemoon, T; Aziz, T; Azzolini, V; Azzopardi, D E; Baak, M A; Back, J J; Bagnasco, S; Bahinipati, S; Bailey, D S; Bailey, S; Bailly, P; van Bakel, N; Bakich, A M; Bala, A; Balagura, V; Baldini-Ferroli, R; Ban, Y; Banas, E; Band, H R; Banerjee, S; Baracchini, E; Barate, R; Barberio, E; Barbero, M; Bard, D J; Barillari, T; Barlow, N R; Barlow, R J; Barrett, M; Bartel, W; Bartelt, J; Bartoldus, R; Batignani, G; Battaglia, M; Bauer, J M; Bay, A; Beaulieu, M; Bechtle, P; Beck, T W; Becker, J; Becla, J; Bedny, I; Behari, S; Behera, P K; Behn, E; Behr, L; Beigbeder, C; Beiline, D; Bell, R; Bellini, F; Bellis, M; Bellodi, G; Belous, K; Benayoun, M; Benelli, G; Benitez, J F; Benkebil, M; Berger, N; Bernabeu, J; Bernard, D; Bernet, R; Bernlochner, F U; Berryhill, J W; Bertsche, K; Besson, P; Best, D S; Bettarini, S; Bettoni, D; Bhardwaj, V; Bhimji, W; Bhuyan, B; Biagini, M E; Biasini, M; van Bibber, K; Biesiada, J; Bingham, I; Bionta, R M; Bischofberger, M; Bitenc, U; Bizjak, I; Blanc, F; Blaylock, G; Blinov, V E; Bloom, E; Bloom, P C; Blount, N L; Blouw, J; Bly, M; Blyth, S; Boeheim, C T; Bomben, M; Bondar, A; Bondioli, M; Bonneaud, G R; Bonvicini, G; Booke, M; Booth, J; Borean, C; Borgland, A W; Borsato, E; Bosi, F; Bosisio, L; Botov, A A; Bougher, J; Bouldin, K; Bourgeois, P; Boutigny, D; Bowerman, D A; Boyarski, A M; Boyce, R F; Boyd, J T; Bozek, A; Bozzi, C; Bračko, M; Brandenburg, G; Brandt, T; Brau, B; Brau, J; Breon, A B; Breton, D; Brew, C; Briand, H; Bright-Thomas, P G; Brigljević, V; Britton, D I; Brochard, F; Broomer, B; Brose, J; Browder, T E; Brown, C L; Brown, C M; Brown, D N; Browne, M; Bruinsma, M; Brunet, S; Bucci, F; Buchanan, C; Buchmueller, O L; Bünger, C; Bugg, W; Bukin, A D; Bula, R; Bulten, H; Burchat, P R; Burgess, W; Burke, J P; Button-Shafer, J; Buzykaev, A R; Buzzo, A; Cai, Y; Calabrese, R; Calcaterra, A; Calderini, G; Camanzi, B; Campagna, E; Campagnari, C; Capra, R; Carassiti, V; Carpinelli, M; Carroll, M; Casarosa, G; Casey, B C K; Cason, N M; Castelli, G; Cavallo, N; Cavoto, G; Cecchi, A; Cenci, R; Cerizza, G; Cervelli, A; Ceseracciu, A; Chai, X; Chaisanguanthum, K S; Chang, M C; Chang, Y H; Chang, Y W; Chao, D S; Chao, M; Chao, Y; Charles, E; Chavez, C A; Cheaib, R; Chekelian, V; Chen, A; Chen, E; Chen, G P; Chen, H F; Chen, J -H; Chen, J C; Chen, K F; Chen, P; Chen, S; Chen, W T; Chen, X; Chen, X R; Chen, Y Q; Cheng, B; Cheon, B G; Chevalier, N; Chia, Y M; Chidzik, S; Chilikin, K; Chistiakova, M V; Cizeron, R; Cho, I S; Cho, K; Chobanova, V; Choi, H H F; Choi, K S; Choi, S K; Choi, Y; Choi, Y K; Christ, S; Chu, P H; Chun, S; Chuvikov, A; Cibinetto, G; Cinabro, D; Clark, A R; Clark, P J; Clarke, C K; Claus, R; Claxton, B; Clifton, Z C; Cochran, J; Cohen-Tanugi, J; Cohn, H; Colberg, T; Cole, S; Colecchia, F; Condurache, C; Contri, R; Convert, P; Convery, M R; Cooke, P; Copty, N; Cormack, C M; Corso, F Dal; Corwin, L A; Cossutti, F; Cote, D; Ramusino, A Cotta; Cottingham, W N; Couderc, F; Coupal, D P; Covarelli, R; Cowan, G; Craddock, W W; Crane, G; Crawley, H B; Cremaldi, L; Crescente, A; Cristinziani, M; Crnkovic, J; Crosetti, G; Cuhadar-Donszelmann, T; Cunha, A; Curry, S; D'Orazio, A; Dû, S; Dahlinger, G; Dahmes, B; Dallapiccola, C; Danielson, N; Danilov, M; Das, A; Dash, M; Dasu, S; Datta, M; Daudo, F; Dauncey, P D; David, P; Davis, C L; Day, C T; De Mori, F; De Domenico, G; De Groot, N; De la Vaissière, C; de la Vaissière, Ch; de Lesquen, A; De Nardo, G; de Sangro, R; De Silva, A; DeBarger, S; Decker, F J; Sanchez, P del Amo; Del Buono, L; Del Gamba, V; del Re, D; Della Ricca, G; Denig, A G; Derkach, D; Derrington, I M; DeStaebler, H; Destree, J; Devmal, S; Dey, B; Di Girolamo, B; Di Marco, E; Dickopp, M; Dima, M O; Dittrich, S; Dittongo, S; Dixon, P; Dneprovsky, L; Dohou, F; Doi, Y; Doležal, Z; Doll, D A; Donald, M; Dong, L; Dong, L Y; Dorfan, J; Dorigo, A; Dorsten, M P; Dowd, R; Dowdell, J; Drásal, Z; Dragic, J; Drummond, B W; Dubitzky, R S; Dubois-Felsmann, G P; Dubrovin, M S; Duh, Y C; Duh, Y T; Dujmic, D; Dungel, W; Dunwoodie, W; Dutta, D; Dvoretskii, A; Dyce, N; Ebert, M; Eckhart, E A; Ecklund, S; Eckmann, R; Eckstein, P; Edgar, C L; Edwards, A J; Egede, U; Eichenbaum, A M; Elmer, P; Emery, S; Enari, Y; Enomoto, R; Erdos, E; Erickson, R; Ernst, J A; Erwin, R J; Escalier, M; Eschenburg, V; Eschrich, I; Esen, S; Esteve, L; Evangelisti, F; Everton, C W; Eyges, V; Fabby, C; Fabozzi, F; Fahey, S; Falbo, M; Fan, S; Fang, F; Fanin, C; Farbin, A; Farhat, H; Fast, J E; Feindt, M; Fella, A; Feltresi, E; Ferber, T; Fernholz, R E; Ferrag, S; Ferrarotto, F; Ferroni, F; Field, R C; Filippi, A; Finocchiaro, G; Fioravanti, E; da Costa, J Firmino; Fischer, P -A; Fisher, A; Fisher, P H; Flacco, C J; Flack, R L; Flaecher, H U; Flanagan, J; Flanigan, J M; Ford, K E; Ford, W T; Forster, I J; Forti, A C; Forti, F; Fortin, D; Foster, B; Foulkes, S D; Fouque, G; Fox, J; Franchini, P; Sevilla, M Franco; Franek, B; Frank, E D; Fransham, K B; Fratina, S; Fratini, K; Frey, A; Frey, R; Friedl, M; Fritsch, M; Fry, J R; Fujii, H; Fujikawa, M; Fujita, Y; Fujiyama, Y; Fukunaga, C; Fukushima, M; Fullwood, J; Funahashi, Y; Funakoshi, Y; Furano, F; Furman, M; Furukawa, K; Futterschneider, H; Gabathuler, E; Gabriel, T A; Gabyshev, N; Gaede, F; Gagliardi, N; Gaidot, A; Gaillard, J -M; Gaillard, J R; Galagedera, S; Galeazzi, F; Gallo, F; Gamba, D; Gamet, R; Gan, K K; Gandini, P; Ganguly, S; Ganzhur, S F; Gao, Y Y; Gaponenko, I; Garmash, A; Tico, J Garra; Garzia, I; Gaspero, M; Gastaldi, F; Gatto, C; Gaur, V; Geddes, N I; Geld, T L; Genat, J -F; George, K A; George, M; George, S; Georgette, Z; Gershon, T J; Gill, M S; Gillard, R; Gilman, J D; Giordano, F; Giorgi, M A; Giraud, P -F; Gladney, L; Glanzman, T; Glattauer, R; Go, A; Goetzen, K; Goh, Y M; Gokhroo, G; Goldenzweig, P; Golubev, V B; Gopal, G P; Gordon, A; Gorišek, A; Goriletsky, V I; Gorodeisky, R; Gosset, L; Gotow, K; Gowdy, S J; Graffin, P; Grancagnolo, S; Grauges, E; Graziani, G; Green, M G; Greene, M G; Grenier, G J; Grenier, P; Griessinger, K; Grillo, A A; Grinyov, B V; Gritsan, A V; Grosdidier, G; Perdekamp, M Grosse; Grosso, P; Grothe, M; Groysman, Y; Grünberg, O; Guido, E; Guler, H; Gunawardane, N J W; Guo, Q H; Guo, R S; Guo, Z J; Guttman, N; Ha, H; Ha, H C; Haas, T; Haba, J; Hachtel, J; Hadavand, H K; Hadig, T; Hagner, C; Haire, M; Haitani, F; Haji, T; Haller, G; Halyo, V; Hamano, K; Hamasaki, H; de Monchenault, G Hamel; Hamilton, J; Hamilton, R; Hamon, O; Han, B Y; Han, Y L; Hanada, H; Hanagaki, K; Handa, F; Hanson, J E; Hanushevsky, A; Hara, K; Hara, T; Harada, Y; Harrison, P F; Harrison, T J; Harrop, B; Hart, A J; Hart, P A; Hartfiel, B L; Harton, J L; Haruyama, T; Hasan, A; Hasegawa, Y; Hast, C; Hastings, N C; Hasuko, K; Hauke, A; Hawkes, C M; Hayashi, K; Hazumi, M; Hee, C; Heenan, E M; Heffernan, D; Held, T; Henderson, R; Henderson, S W; Hertzbach, S S; Hervé, S; Heß, M; Heusch, C A; Hicheur, A; Higashi, Y; Higasino, Y; Higuchi, I; Hikita, S; Hill, E J; Himel, T; Hinz, L; Hirai, T; Hirano, H; Hirschauer, J F; Hitlin, D G; Hitomi, N; Hodgkinson, M C; Höcker, A; Hoi, C T; Hojo, T; Hokuue, T; Hollar, J J; Hong, T M; Honscheid, K; Hooberman, B; Hopkins, D A; Horii, Y; Hoshi, Y; Hoshina, K; Hou, S; Hou, W S; Hryn'ova, T; Hsiung, Y B; Hsu, C L; Hsu, S C; Hu, H; Hu, T; Huang, H C; Huang, T J; Huang, Y C; Huard, Z; Huffer, M E; Hufnagel, D; Hung, T; Hutchcroft, D E; Hyun, H J; Ichizawa, S; Igaki, T; Igarashi, A; Igarashi, S; Igarashi, Y; Igonkina, O; Ikado, K; Ikeda, H; Ikeda, K; Ilic, J; Inami, K; Innes, W R; Inoue, Y; Ishikawa, A; Ishino, H; Itagaki, K; Itami, S; Itoh, K; Ivanchenko, V N; Iverson, R; Iwabuchi, M; Iwai, G; Iwai, M; Iwaida, S; Iwamoto, M; Iwasaki, H; Iwasaki, M; Iwashita, T; Izen, J M; Jackson, D J; Jackson, F; Jackson, G; Jackson, P S; Jacobsen, R G; Jacoby, C; Jaegle, I; Jain, V; Jalocha, P; Jang, H K; Jasper, H; Jawahery, A; Jayatilleke, S; Jen, C M; Jensen, F; Jessop, C P; Ji, X B; John, M J J; Johnson, D R; Johnson, J R; Jolly, S; Jones, M; Joo, K K; Joshi, N; Joshi, N J; Judd, D; Julius, T; Kadel, R W; Kadyk, J A; Kagan, H; Kagan, R; Kah, D H; Kaiser, S; Kaji, H; Kajiwara, S; Kakuno, H; Kameshima, T; Kaminski, J; Kamitani, T; Kaneko, J; Kang, J H; Kang, J S; Kani, T; Kapusta, P; Karbach, T M; Karolak, M; Karyotakis, Y; Kasami, K; Katano, G; Kataoka, S U; Katayama, N; Kato, E; Kato, Y; Kawai, H; Kawai, M; Kawamura, N; Kawasaki, T; Kay, J; Kay, M; Kelly, M P; Kelsey, M H; Kent, N; Kerth, L T; Khan, A; Khan, H R; Kharakh, D; Kibayashi, A; Kichimi, H; Kiesling, C; Kikuchi, M; Kikutani, E; Kim, B H; Kim, C H; Kim, D W; Kim, H; Kim, H J; Kim, H O; Kim, H W; Kim, J B; Kim, J H; Kim, K T; Kim, M J; Kim, P; Kim, S K; Kim, S M; Kim, T H; Kim, Y I; Kim, Y J; King, G J; Kinoshita, K; Kirk, A; Kirkby, D; Kitayama, I; Klemetti, M; Klose, V; Klucar, J; Knecht, N S; Knoepfel, K J; Knowles, D J; Ko, B R; Kobayashi, N; Kobayashi, S; Kobayashi, T; Kobel, M J; Koblitz, S; Koch, H; Kocian, M L; Kodyš, P; Koeneke, K; Kofler, R; Koike, S; Koishi, S; Koiso, H; Kolb, J A; Kolya, S D; Kondo, Y; Konishi, H; Koppenburg, P; Koptchev, V B; Kordich, T M B; Korol, A A; Korotushenko, K; Korpar, S; Kouzes, R T; Kovalskyi, D; Kowalewski, R; Kozakai, Y; Kozanecki, W; Kral, J F; Krasnykh, A; Krause, R; Kravchenko, E A; Krebs, J; Kreisel, A; Kreps, M; Krishnamurthy, M; Kroeger, R; Kroeger, W; Krokovny, P; Kronenbitter, B; Kroseberg, J; Kubo, T; Kuhr, T; Kukartsev, G; Kulasiri, R; Kulikov, A; Kumar, R; Kumar, S; Kumita, T; Kuniya, T; Kunze, M; Kuo, C C; Kuo, T -L; Kurashiro, H; Kurihara, E; Kurita, N; Kuroki, Y; Kurup, A; Kutter, P E; Kuznetsova, N; Kvasnička, P; Kyberd, P; Kyeong, S H; Lacker, H M; Lae, C K; Lamanna, E; Lamsa, J; Lanceri, L; Landi, L; Lang, M I; Lange, D J; Lange, J S; Langenegger, U; Langer, M; Lankford, A J; Lanni, F; Laplace, S; Latour, E; Lau, Y P; Lavin, D R; Layter, J; Lebbolo, H; LeClerc, C; Leddig, T; Leder, G; Diberder, F Le; Lee, C L; Lee, J; Lee, J S; Lee, M C; Lee, M H; Lee, M J; Lee, S -J; Lee, S E; Lee, S H; Lee, Y J; Lees, J P; Legendre, M; Leitgab, M; Leitner, R; Leonardi, E; Leonidopoulos, C; Lepeltier, V; Leruste, Ph; Lesiak, T; Levi, M E; Levy, S L; Lewandowski, B; Lewczuk, M J; Lewis, P; Li, H; Li, H B; Li, S; Li, X; Li, Y; Gioi, L Li; Libby, J; Lidbury, J; Lillard, V; Lim, C L; Limosani, A; Lin, C S; Lin, J Y; Lin, S W; Lin, Y S; Lindquist, B; Lindsay, C; Lista, L; Liu, C; Liu, F; Liu, H; Liu, H M; Liu, J; Liu, R; Liu, T; Liu, Y; Liu, Z Q; Liventsev, D; Vetere, M Lo; Locke, C B; Lockman, W S; Di Lodovico, F; Lombardo, V; London, G W; Pegna, D Lopes; Lopez, L; Lopez-March, N; Lory, J; LoSecco, J M; Lou, X C; Louvot, R; Lu, A; Lu, C; Lu, M; Lu, R S; Lueck, T; Luitz, S; Lukin, P; Lund, P; Luppi, E; Lutz, A M; Lutz, O; Lynch, G; Lynch, H L; Lyon, A J; Lyubinsky, V R; MacFarlane, D B; Mackay, C; MacNaughton, J; Macri, M M; Madani, S; Mader, W F; Majewski, S A; Majumder, G; Makida, Y; Malaescu, B; Malaguti, R; Malclès, J; Mallik, U; Maly, E; Mamada, H; Manabe, A; Mancinelli, G; Mandelkern, M; Mandl, F; Manfredi, P F; Mangeol, D J J; Manoni, E; Mao, Z P; Margoni, M; Marker, C E; Markey, G; Marks, J; Marlow, D; Marques, V; Marsiske, H; Martellotti, S; Martin, E C; Martin, J P; Martin, L; Martinez, A J; Marzolla, M; Mass, A; Masuzawa, M; Mathieu, A; Matricon, P; Matsubara, T; Matsuda, T; Matsumoto, H; Matsumoto, S; Matsumoto, T; Matsuo, H; Mattison, T S; Matvienko, D; Matyja, A; Mayer, B; Mazur, M A; Mazzoni, M A; McCulloch, M; McDonald, J; McFall, J D; McGrath, P; McKemey, A K; McKenna, J A; Mclachlin, S E; McMahon, S; McMahon, T R; McOnie, S; Medvedeva, T; Melen, R; Mellado, B; Menges, W; Menke, S; Merchant, A M; Merkel, J; Messner, R; Metcalfe, S; Metzler, S; Meyer, N T; Meyer, T I; Meyer, W T; Michael, A K; Michelon, G; Michizono, S; Micout, P; Miftakov, V; Mihalyi, A; Mikami, Y; Milanes, D A; Milek, M; Mimashi, T; Minamora, J S; Mindas, C; Minutoli, S; Mir, L M; Mishra, K; Mitaroff, W; Miyake, H; Miyashita, T S; Miyata, H; Miyazaki, Y; Moffitt, L C; Mohapatra, A; Mohapatra, A K; Mohapatra, D; Moll, A; Moloney, G R; Mols, J P; Mommsen, R K; Monge, M R; Monorchio, D; Moore, T B; Moorhead, G F; de Freitas, P Mora; Morandin, M; Morgan, N; Morgan, S E; Morganti, M; Morganti, S; Mori, S; Mori, T; Morii, M; Morris, J P; Morsani, F; Morton, G W; Moss, L J; Mouly, J P; Mount, R; Mueller, J; Müller-Pfefferkorn, R; Mugge, M; Muheim, F; Muir, A; Mullin, E; Munerato, M; Murakami, A; Murakami, T; Muramatsu, N; Musico, P; Nagai, I; Nagamine, T; Nagasaka, Y; Nagashima, Y; Nagayama, S; Nagel, M; Naisbit, M T; Nakadaira, T; Nakahama, Y; Nakajima, M; Nakajima, T; Nakamura, I; Nakamura, T; Nakamura, T T; Nakano, E; Nakayama, H; Nam, J W; Narita, S; Narsky, I; Nash, J A; Natkaniec, Z; Nauenberg, U; Nayak, M; Neal, H; Nedelkovska, E; Negrini, M; Neichi, K; Nelson, D; Nelson, S; Neri, N; Nesom, G; Neubauer, S; Newman-Coburn, D; Ng, C; Nguyen, X; Nicholson, H; Niebuhr, C; Nief, J Y; Niiyama, M; Nikolich, M B; Nisar, N K; Nishimura, K; Nishio, Y; Nitoh, O; Nogowski, R; Noguchi, S; Nomura, T; Nordby, M; Nosochkov, Y; Novokhatski, A; Nozaki, S; Nozaki, T; Nugent, I M; O'Grady, C P; O'Neale, S W; O'Neill, F G; Oberhof, B; Oddone, P J; Ofte, I; Ogawa, A; Ogawa, K; Ogawa, S; Ogawa, Y; Ohkubo, R; Ohmi, K; Ohnishi, Y; Ohno, F; Ohshima, T; Ohshima, Y; Ohuchi, N; Oide, K; Oishi, N; Okabe, T; Okazaki, N; Okazaki, T; Okuno, S; Olaiya, E O; Olivas, A; Olley, P; Olsen, J; Ono, S; Onorato, G; Onuchin, A P; Onuki, Y; Ooba, T; Orimoto, T J; Oshima, T; Osipenkov, I L; Ostrowicz, W; Oswald, C; Otto, S; Oyang, J; Oyanguren, A; Ozaki, H; Ozcan, V E; Paar, H P; Padoan, C; Paick, K; Palka, H; Pan, B; Pan, Y; Vazquez, W Panduro; Panetta, J; Panova, A I; Panvini, R S; Panzenböck, E; Paoloni, E; Paolucci, P; Pappagallo, M; Paramesvaran, S; Park, C S; Park, C W; Park, H; Park, H K; Park, K S; Park, W; Parry, R J; Parslow, N; Passaggio, S; Pastore, F C; Patel, P M; Patrignani, C; Patteri, P; Pavel, T; Pavlovich, J; Payne, D J; Peak, L S; Peimer, D R; Pelizaeus, M; Pellegrini, R; Pelliccioni, M; Peng, C C; Peng, J C; Peng, K C; Peng, T; Penichot, Y; Pennazzi, S; Pennington, M R; Penny, R C; Penzkofer, A; Perazzo, A; Perez, A; Perl, M; Pernicka, M; Perroud, J -P; Peruzzi, I M; Pestotnik, R; Peters, K; Peters, M; Petersen, B A; Petersen, T C; Petigura, E; Petrak, S; Petrella, A; Petrič, M; Petzold, A; Pia, M G; Piatenko, T; Piccolo, D; Piccolo, M; Piemontese, L; Piemontese, M; Pierini, M; Pierson, S; Pioppi, M; Piredda, G; Pivk, M; Plaszczynski, S; Polci, F; Pompili, A; Poropat, P; Posocco, M; Potter, C T; Potter, R J L; Prasad, V; Prebys, E; Prencipe, E; Prendki, J; Prepost, R; Prest, M; Prim, M; Pripstein, M; Prudent, X; Pruvot, S; Puccio, E M T; Purohit, M V; Qi, N D; Quinn, H; Raaf, J; Rabberman, R; Raffaelli, F; Ragghianti, G; Rahatlou, S; Rahimi, A M; Rahmat, R; Rakitin, A Y; Randle-Conde, A; Rankin, P; Rashevskaya, I; Ratkovsky, S; Raven, G; Re, V; Reep, M; Regensburger, J J; Reidy, J; Reif, R; Reisert, B; Renard, C; Renga, F; Ricciardi, S; Richman, J D; Ritchie, J L; Ritter, M; Rivetta, C; Rizzo, G; Roat, C; Robbe, P; Roberts, D A; Robertson, A I; Robutti, E; Rodier, S; Rodriguez, D M; Rodriguez, J L; Rodriguez, R; Roe, N A; Röhrken, M; Roethel, W; Rolquin, J; Romanov, L; Romosan, A; Ronan, M T; Rong, G; Ronga, F J; Roos, L; Root, N; Rosen, M; Rosenberg, E I; Rossi, A; Rostomyan, A; Rotondo, M; Roussot, E; Roy, J; Rozanska, M; Rozen, Y; Rubin, A E; Ruddick, W O; Ruland, A M; Rybicki, K; Ryd, A; Ryu, S; Ryuko, J; Sabik, S; Sacco, R; Saeed, M A; Tehrani, F Safai; Sagawa, H; Sahoo, H; Sahu, S; Saigo, M; Saito, T; Saitoh, S; Sakai, K; Sakamoto, H; Sakaue, H; Saleem, M; Salnikov, A A; Salvati, E; Salvatore, F; Samuel, A; Sanders, D A; Sanders, P; Sandilya, S; Sandrelli, F; Sands, W; Sands, W R; Sanpei, M; Santel, D; Santelj, L; Santoro, V; Santroni, A; Sanuki, T; Sarangi, T R; Saremi, S; Sarti, A; Sasaki, T; Sasao, N; Satapathy, M; Sato, Nobuhiko; Sato, Noriaki; Sato, Y; Satoyama, N; Satpathy, A; Savinov, V; Savvas, N; Saxton, O H; Sayeed, K; Schaffner, S F; Schalk, T; Schenk, S; Schieck, J R; Schietinger, T; Schilling, C J; Schindler, R H; Schmid, S; Schmitz, R E; Schmuecker, H; Schneider, O; Schnell, G; Schönmeier, P; Schofield, K C; Schott, G; Schröder, H; Schram, M; Schubert, J; Schümann, J; Schultz, J; Schumm, B A; Schune, M H; Schwanke, U; Schwarz, H; Schwiening, J; Schwierz, R; Schwitters, R F; Sciacca, C; Sciolla, G; Scott, I J; Seeman, J; Seiden, A; Seitz, R; Seki, T; Sekiya, A I; Semenov, S; Semmler, D; Sen, S; Senyo, K; Seon, O; Serbo, V V; Serednyakov, S I; Serfass, B; Serra, M; Serrano, J; Settai, Y; Seuster, R; Sevior, M E; Shakhova, K V; Shang, L; Shapkin, M; Sharma, V; Shebalin, V; Shelkov, V G; Shen, B C; Shen, D Z; Shen, Y T; Sherwood, D J; Shibata, T; Shibata, T A; Shibuya, H; Shidara, T; Shimada, K; Shimoyama, M; Shinomiya, S; Shiu, J G; Shorthouse, H W; Shpilinskaya, L I; Sibidanov, A; Sicard, E; Sidorov, A; Sidorov, V; Siegle, V; Sigamani, M; Simani, M C; Simard, M; Simi, G; Simon, F; Simonetto, F; Sinev, N B; Singh, H; Singh, J B; Sinha, R; Sitt, S; Skovpen, Yu I; Sloane, R J; Smerkol, P; Smith, A J S; Smith, D; Smith, D S; Smith, J G; Smol, A; Snoek, H L; Snyder, A; So, R Y; Sobie, R J; Soderstrom, E; Soha, A; Sohn, Y S; Sokoloff, M D; Sokolov, A; Solagna, P; Solovieva, E; Soni, N; Sonnek, P; Sordini, V; Spaan, B; Spanier, S M; Spencer, E; Speziali, V; Spitznagel, M; Spradlin, P; Staengle, H; Stamen, R; Stanek, M; Stanič, S; Stark, J; Steder, M; Steininger, H; Steinke, M; Stelzer, J; Stevanato, E; Stocchi, A; Stock, R; Stoeck, H; Stoker, D P; Stroili, R; Strom, D; Strother, P; Strube, J; Stugu, B; Stypula, J; Su, D; Suda, R; Sugahara, R; Sugi, A; Sugimura, T; Sugiyama, A; Suitoh, S; Sullivan, M K; Sumihama, M; Sumiyoshi, T; Summers, D J; Sun, L; Sun, S; Sundermann, J E; Sung, H F; Susaki, Y; Sutcliffe, P; Suzuki, A; Suzuki, J; Suzuki, J I; Suzuki, K; Suzuki, S; Suzuki, S Y; Swain, J E; Swain, S K; T'Jampens, S; Tabata, M; Tackmann, K; Tajima, H; Tajima, O; Takahashi, K; Takahashi, S; Takahashi, T; Takasaki, F; Takayama, T; Takita, M; Tamai, K; Tamponi, U; Tamura, N; Tan, N; Tan, P; Tanabe, K; Tanabe, T; Tanaka, H A; Tanaka, J; Tanaka, M; Tanaka, S; Tanaka, Y; Tanida, K; Taniguchi, N; Taras, P; Tasneem, N; Tatishvili, G; Tatomi, T; Tawada, M; Taylor, F; Taylor, G N; Taylor, G P; Telnov, V I; Teodorescu, L; Ter-Antonyan, R; Teramoto, Y; Teytelman, D; Thérin, G; Thiebaux, Ch; Thiessen, D; Thomas, E W; Thompson, J M; Thorne, F; Tian, X C; Tibbetts, M; Tikhomirov, I; Tinslay, J S; Tiozzo, G; Tisserand, V; Tocut, V; Toki, W H; Tomassini, E W; Tomoto, M; Tomura, T; Torassa, E; Torrence, E; Tosi, S; Touramanis, C; Toussaint, J C; Tovey, S N; Trapani, P P; Treadwell, E; Triggiani, G; Trincaz-Duvoid, S; Trischuk, W; Troost, D; Trunov, A; Tsai, K L; Tsai, Y T; Tsujita, Y; Tsukada, K; Tsukamoto, T; Tuggle, J M; Tumanov, A; Tung, Y W; Turnbull, L; Turner, J; Turri, M; Uchida, K; Uchida, M; Uchida, Y; Ueki, M; Ueno, K; Ujiie, N; Ulmer, K A; Unno, Y; Urquijo, P; Ushiroda, Y; Usov, Y; Usseglio, M; Usuki, Y; Uwer, U; Va'vra, J; Vahsen, S E; Vaitsas, G; Valassi, A; Vallazza, E; Vallereau, A; Vanhoefer, P; van Hoek, W C; Van Hulse, C; van Winkle, D; Varner, G; Varnes, E W; Varvell, K E; Vasileiadis, G; Velikzhanin, Y S; Verderi, M; Versillé, S; Vervink, K; Viaud, B; Vidal, P B; Villa, S; Villanueva-Perez, P; Vinograd, E L; Vitale, L; Vitug, G M; Voß, C; Voci, C; Voena, C; Volk, A; von Wimmersperg-Toeller, J H; Vorobyev, V; Vossen, A; Vuagnin, G; Vuosalo, C O; Wacker, K; Wagner, A P; Wagner, D L; Wagner, G; Wagner, M N; Wagner, S R; Wagoner, D E; Walker, D; Walkowiak, W; Wallom, D; Wang, C C; Wang, C H; Wang, J; Wang, J G; Wang, K; Wang, L; Wang, L L; Wang, P; Wang, T J; Wang, W F; Wang, X L; Wang, Y F; Wappler, F R; Watanabe, M; Watson, A T; Watson, J E; Watson, N K; Watt, M; Weatherall, J H; Weaver, M; Weber, T; Wedd, R; Wei, J T; Weidemann, A W; W, A J R; Wenzel, W A; West, C A; West, C G; West, T J; White, R M; Wicht, J; Widhalm, L; Wiechczynski, J; Wienands, U; Wilden, L; Wilder, M; Williams, D C; Williams, G; Williams, J C; Williams, K M; Williams, M I; Willocq, S Y; Wilson, J R; Wilson, M G; Wilson, R J; Winklmeier, F; Winter, M A; Wisniewski, W J; Wittgen, M; Wittlin, J; Wittmer, W; Wixted, R; Woch, A; Wogsland, B J; Wong, Q K; Wray, B C; Wren, A C; Wright, D M; Wu, C H; Wu, J; Wu, S L; Wulsin, H W; Xella, S M; Xie, Q L; Xie, Y; Xu, Z Z; Yèche, Ch; Yamada, Y; Yamaga, M; Yamaguchi, A; Yamaguchi, H; Yamaki, T; Yamamoto, H; Yamamoto, N; Yamamoto, R K; Yamamoto, S; Yamanaka, T; Yamaoka, H; Yamaoka, J; Yamaoka, Y; Yamashita, Y; Yamauchi, M; Yan, D S; Yan, Y; Yanai, H; Yanaka, S; Yang, H; Yang, R; Yang, S; Yarritu, A K; Yashchenko, S; Yashima, J; Yasin, Z; Yasu, Y; Ye, S W; Yeh, P; Yi, J I; Yi, K; Yi, M; Yin, Z W; Ying, J; Yocky, G; Yokoyama, K; Yokoyama, M; Yokoyama, T; Yoshida, K; Yoshida, M; Yoshimura, Y; Young, C C; Yu, C X; Yu, Z; Yuan, C Z; Yuan, Y; Yumiceva, F X; Yusa, Y; Yushkov, A N; Yuta, H; Zacek, V; Zain, S B; Zallo, A; Zambito, S; Zander, D; Zang, S L; Zanin, D; Zaslavsky, B G; Zeng, Q L; Zghiche, A; Zhang, B; Zhang, J; Zhang, L; Zhang, L M; Zhang, S Q; Zhang, Z P; Zhao, H W; Zhao, M; Zhao, Z G; Zheng, Y; Zheng, Y H; Zheng, Z P; Zhilich, V; Zhou, P; Zhu, R Y; Zhu, Y S; Zhu, Z M; Zhulanov, V; Ziegler, T; Ziegler, V; Zioulas, G; Zisman, M; Zito, M; Zürcher, D; Zwahlen, N; Zyukova, O; Živko, T; Žontar, D; Bevan, Adrian; Golob, Bostjan; Mannel, Thomas; Prell, Soeren; Yabsley, Bruce

    2014-01-01

    This work is on the Physics of the B Factories. Part A of this book contains a brief description of the SLAC and KEK B Factories as well as their detectors, BaBar and Belle, and data taking related issues. Part B discusses tools and methods used by the experiments in order to obtain results. The results themselves can be found in Part C.

  19. Aspergillus luchuensis, an industrially important black Aspergillus in East Asia.

    Directory of Open Access Journals (Sweden)

    Seung-Beom Hong

    Full Text Available Aspergilli known as black- and white-koji molds which are used for awamori, shochu, makgeolli and other food and beverage fermentations, are reported in the literature as A. luchuensis, A. awamori, A. kawachii, or A. acidus. In order to elucidate the taxonomic position of these species, available ex-type cultures were compared based on morphology and molecular characters. A. luchuensis, A. kawachii and A. acidus showed the same banding patterns in RAPD, and the three species had the same rDNA-ITS, β-tubulin and calmodulin sequences and these differed from those of the closely related A. niger and A. tubingensis. Morphologically, the three species are not significantly different from each other or from A. niger and A. tubingensis. It is concluded that A. luchuensis, A. kawachii and A. acidus are the same species, and A. luchuensis is selected as the correct name based on priority. Strains of A. awamori which are stored in National Research Institute of Brewing in Japan, represent A. niger (n = 14 and A. luchuensis (n = 6. The neotype of A. awamori (CBS 557.65 =  NRRL 4948 does not originate from awamori fermentation and it is shown to be identical with the unknown taxon Aspergillus welwitschiae. Extrolite analysis of strains of A. luchuensis showed that they do not produce mycotoxins and therefore can be considered safe for food and beverage fermentations. A. luchuensis is also frequently isolated from meju and nuruk in Korea and Puerh tea in China and the species is probably common in the fermentation environment of East Asia. A re-description of A. luchuensis is provided because the incomplete data in the original literature.

  20. New promoters to improve heterologous protein production in Aspergillus vadensis

    NARCIS (Netherlands)

    Culleton, H.; Bouzid, O.; McKie, V.; de Vries, Ronald P.

    2014-01-01

    Aspergillus is a widely used host organism for the industrial production of homologous and heterologous proteins. Although Aspergillus niger is most commonly used, a close relative of this species, Aspergillus vadensis, has been suggested as a suitable and more favourable alternative due in part, to

  1. 21 CFR 866.3040 - Aspergillus spp. serological reagents.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Aspergillus spp. serological reagents. 866.3040... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3040 Aspergillus spp. serological reagents. (a) Identification. Aspergillus spp. serological reagents are devices...

  2. [Aspergillus fumigatus endocarditis in a patient with a biventricular pacemaker].

    Science.gov (United States)

    Cuesta, José M; Fariñas, María C; Rodilla, Irene G; Salesa, Ricardo; de Berrazueta, José R

    2005-05-01

    Aspergillus fumigatus endocarditis is one of the rarest and severest complications in cardiological patients. We describe a patient with an intracardial pacemaker who was diagnosed as having Aspergillus fumigatus endocarditis. Postmortem examination showed a large, Aspergillus-infected thrombus encased in the right ventricle, pulmonary trunk and main pulmonary branches.

  3. Aspergillus vertebral osteomyelitis in immunocompetent patients

    Directory of Open Access Journals (Sweden)

    Somika Sethi

    2012-01-01

    Full Text Available Fungal infections are one of the important cause of morbidity and mortality in immunocompromised patients. Aspergillus vertebral osteomyelitis is extremely rare. We report two cases of aspergillus vertebral osteomyelitis in immunocompetent men in the absence of an underlying disorder. The clinical and radiological findings were suggestive of Pott′s spine. The absolute CD4, CD8 counts and their ratio were normal. The HIV status was negative in both patients. Both patients underwent surgical decompression. The histopathology of tissue obtained were suggestive of aspergillus osteomyelitis. One patient had antifungal treatment for 3 months and was doing well at 1 year followup, whereas other patient did not turnup after 2 months.

  4. A rare case of bilateral aspergillus endophthalmitis

    Directory of Open Access Journals (Sweden)

    Saurabh Gupta

    2015-12-01

    Full Text Available Aspergillus endophthalmitis is a devastating inflammatory condition of the intraocular cavities that may result in irreparable loss of vision and rapid destruction of the eye. Almost all cases in the literature have shown an identified source causing aspergillus endophthalmitis as a result of direct extension of disease. We present a rare case of bilateral aspergillus endophthalmitis. A 72-year-old woman with a history of diabetes mellitus, congenital Hirschsprung disease, and recent culture-positive candida pyelonephritis with hydronephrosis status post-surgical stent placement presented with difficulty opening her eyes. She complained of decreased vision (20/200 with pain and redness in both eyes – right worse then left. Examination demonstrated multiple white fungal balls in both retinas consistent with bilateral fungal endophthalmitis. Bilateral vitreous taps for cultures and staining were performed. Patient was given intravitreal injections of amphotericin B, vancomycin, ceftazidime, and started on oral fluconazole. Patient was scheduled for vitrectomy to decrease organism burden and to remove loculated areas of infection that would not respond to systemic antifungal agents. Four weeks after initial presentation, the fungal cultures revealed mold growth consistent with aspergillus. Patient was subsequently started on voriconazole and fluconazole was discontinued due to poor efficacy against aspergillus. Further workup was conducted to evaluate for the source of infection and seeding. Transthoracic cardiogram was unremarkable for any vegetation or valvular abnormalities. MRI of the orbits and sinuses did not reveal any mass lesions or bony destruction. CT of the chest was unremarkable for infection. Aspergillus endophthalmitis may occur because of one of these several mechanisms: hematogenous dissemination, direct inoculation by trauma, and contamination during surgery. Our patient's cause of bilateral endophthalmitis was through an

  5. Evaluation of antifungal activity of essential oils against potentially mycotoxigenic Aspergillus flavus and Aspergillus parasiticus

    Directory of Open Access Journals (Sweden)

    Fernanda C. da Silva

    2012-10-01

    Full Text Available The antifungal activity of essential oils of fennel (Foeniculum vulgare Mill., Apiaceae, ginger (Zingiber officinale Roscoe, Zingiberaceae, mint (Mentha piperita L., Lamiaceae and thyme (Thymus vulgaris L., Lamiaceae was evaluated against mycotoxin producers Aspergillus flavus and A. parasiticus. High Resolution Gas Chromatography was applied to analyze chemical constituents of essential oils. The effect of different concentrations of essential oils was determined by solid medium diffusion assay. Mycelial growth and sporulation were determined for each essential oil at the concentrations established by solid medium diffusion assay. At the fifth, seventh and ninth days the mycelial diameter (Ø mm and spore production were also determined. FUN-1 staining was performed to assess cell viability after broth macrodilution assay. Trans-anethole, zingiberene, menthol and thymol are the major component of essential oils of fennel, ginger, mint and thyme, respectively. The effective concentrations for fennel, ginger, mint and thyme were 50, 80, 50 and 50% (oil/DMSO; v/v, respectively. The four essential oils analysed in this study showed antifungal effect. Additionally, FUN-1 staining showed to be a suitable method to evaluate cell viability of potential mycotoxigenic fungi A. flavus and A. parasiticus after treatment with essential oils.

  6. Aspergillus species cystitis in a cat.

    Science.gov (United States)

    Adamama-Moraitou, K K; Paitaki, C G; Rallis, T S; Tontis, D

    2001-03-01

    A Persian male cat with a history of lower urinary tract disease was presented because of polydipsia, polyuria, constipation and nasal discharge. Ten weeks before admission, the cat had been treated for lower urinary tract disease by catheterisation and flushing of the bladder. The animal was thin, dehydrated, anaemic and azotaemic. Urine culture revealed Aspergillus species cystitis. Antibodies against Aspergillus nidulans were identified in serum. Fluconazole was administered orally (7.5 mg/kg, q 12 h) for 10 consecutive weeks. The azotaemia was resolved, the kidney concentrating ability was recovered and the cat has remained healthy without similar problems.

  7. Aspergillus bertholletius sp. nov. from Brazil Nuts

    DEFF Research Database (Denmark)

    Taniwaki, Marta H.; Pitt, John I.; Iamanaka, Beatriz T.

    2012-01-01

    During a study on the mycobiota of brazil nuts (Bertholletia excelsa) in Brazil, a new Aspergillus species, A. bertholletius, was found, and is described here. A polyphasic approach was applied using morphological characters, extrolite data as well as partial beta-tubulin, calmodulin and ITS...... acid and ustilaginoidin C. Phylogenetic analysis using partial beta-tubulin and camodulin gene sequences showed that A. bertholletius represents a new phylogenetic clade in Aspergillus section Flavi. The type strain of A. bertholletius is CCT 7615 (=ITAL 270/06 = IBT 29228)....

  8. 27 CFR 40.254 - Receipt into factory.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 2 2010-04-01 2010-04-01 false Receipt into factory. 40... Operations § 40.254 Receipt into factory. A manufacturer of tobacco products may receive in bond into his factory tobacco products and may also receive into his factory tobacco products on which the tax has...

  9. Functional analysis of the Aspergillus nidulans kinome.

    Science.gov (United States)

    De Souza, Colin P; Hashmi, Shahr B; Osmani, Aysha H; Andrews, Peter; Ringelberg, Carol S; Dunlap, Jay C; Osmani, Stephen A

    2013-01-01

    The filamentous fungi are an ecologically important group of organisms which also have important industrial applications but devastating effects as pathogens and agents of food spoilage. Protein kinases have been implicated in the regulation of virtually all biological processes but how they regulate filamentous fungal specific processes is not understood. The filamentous fungus Aspergillus nidulans has long been utilized as a powerful molecular genetic system and recent technical advances have made systematic approaches to study large gene sets possible. To enhance A. nidulans functional genomics we have created gene deletion constructs for 9851 genes representing 93.3% of the encoding genome. To illustrate the utility of these constructs, and advance the understanding of fungal kinases, we have systematically generated deletion strains for 128 A. nidulans kinases including expanded groups of 15 histidine kinases, 7 SRPK (serine-arginine protein kinases) kinases and an interesting group of 11 filamentous fungal specific kinases. We defined the terminal phenotype of 23 of the 25 essential kinases by heterokaryon rescue and identified phenotypes for 43 of the 103 non-essential kinases. Uncovered phenotypes ranged from almost no growth for a small number of essential kinases implicated in processes such as ribosomal biosynthesis, to conditional defects in response to cellular stresses. The data provide experimental evidence that previously uncharacterized kinases function in the septation initiation network, the cell wall integrity and the morphogenesis Orb6 kinase signaling pathways, as well as in pathways regulating vesicular trafficking, sexual development and secondary metabolism. Finally, we identify ChkC as a third effector kinase functioning in the cellular response to genotoxic stress. The identification of many previously unknown functions for kinases through the functional analysis of the A. nidulans kinome illustrates the utility of the A. nidulans gene

  10. Functional analysis of the Aspergillus nidulans kinome.

    Directory of Open Access Journals (Sweden)

    Colin P De Souza

    Full Text Available The filamentous fungi are an ecologically important group of organisms which also have important industrial applications but devastating effects as pathogens and agents of food spoilage. Protein kinases have been implicated in the regulation of virtually all biological processes but how they regulate filamentous fungal specific processes is not understood. The filamentous fungus Aspergillus nidulans has long been utilized as a powerful molecular genetic system and recent technical advances have made systematic approaches to study large gene sets possible. To enhance A. nidulans functional genomics we have created gene deletion constructs for 9851 genes representing 93.3% of the encoding genome. To illustrate the utility of these constructs, and advance the understanding of fungal kinases, we have systematically generated deletion strains for 128 A. nidulans kinases including expanded groups of 15 histidine kinases, 7 SRPK (serine-arginine protein kinases kinases and an interesting group of 11 filamentous fungal specific kinases. We defined the terminal phenotype of 23 of the 25 essential kinases by heterokaryon rescue and identified phenotypes for 43 of the 103 non-essential kinases. Uncovered phenotypes ranged from almost no growth for a small number of essential kinases implicated in processes such as ribosomal biosynthesis, to conditional defects in response to cellular stresses. The data provide experimental evidence that previously uncharacterized kinases function in the septation initiation network, the cell wall integrity and the morphogenesis Orb6 kinase signaling pathways, as well as in pathways regulating vesicular trafficking, sexual development and secondary metabolism. Finally, we identify ChkC as a third effector kinase functioning in the cellular response to genotoxic stress. The identification of many previously unknown functions for kinases through the functional analysis of the A. nidulans kinome illustrates the utility of the

  11. Factorials of real negative and imaginary numbers - A new perspective.

    Science.gov (United States)

    Thukral, Ashwani K

    2014-01-01

    Presently, factorials of real negative numbers and imaginary numbers, except for zero and negative integers are interpolated using the Euler's gamma function. In the present paper, the concept of factorials has been generalised as applicable to real and imaginary numbers, and multifactorials. New functions based on Euler's factorial function have been proposed for the factorials of real negative and imaginary numbers. As per the present concept, the factorials of real negative numbers, are complex numbers. The factorials of real negative integers have their imaginary part equal to zero, thus are real numbers. Similarly, the factorials of imaginary numbers are complex numbers. The moduli of the complex factorials of real negative numbers, and imaginary numbers are equal to their respective real positive number factorials. Fractional factorials and multifactorials have been defined in a new perspective. The proposed concept has also been extended to Euler's gamma function for real negative numbers and imaginary numbers, and beta function.

  12. Aspergillus brasiliensis sp. nov., a biseriate black Aspergillus species with world-wide distribution

    DEFF Research Database (Denmark)

    Varga, János; Kocsubé, Sándor; Tóth, Beáta

    2007-01-01

    A novel species, Aspergillus brasiliensis sp. nov., is described within Aspergillus section Nigri. This species can be distinguished from other black aspergilli based on intergenic transcribed region, beta-tubulin and calmodulin gene sequences, by amplified fragment length polymorphism analysis...... and by extrolite profiles. A. brasiliensis isolates produced naphtho-gamma-pyrones, tensidol A and B and pyrophen in common with Aspergiflus niger and Aspergillus tubingensis, but also several unique compounds, justifying their treatment as representing a separate species. None of the isolates were found...... to produce ochratoxin A, kotanins, funalenone or pyranonigrins. The novel species was most closely related to A. niger, and was isolated from soil from Brazil, Australia, USA and The Netherlands, and from grape berries from Portugal. The type strain of Aspergillus brasiliensis sp. nov. is CBS 101740(T) (=IM...

  13. Density and molecular epidemiology of Aspergillus in air and relationship to outbreaks of Aspergillus infection

    NARCIS (Netherlands)

    A.C.A.P. Leenders (Alexander); A.F. van Belkum (Alex); M.D. Behrendt (Myra); A. Luijendijk (Ad); H.A. Verbrugh (Henri)

    1999-01-01

    textabstractAfter five patients were diagnosed with nosocomial invasive aspergillosis caused by Aspergillus fumigatus and A. flavus, a 14-month surveillance program for pathogenic and nonpathogenic fungal conidia in the air within and outside the University Hospital in

  14. A Saccharomyces cerevisiae Wine Strain Inhibits Growth and Decreases Ochratoxin A Biosynthesis by Aspergillus carbonarius and Aspergillus ochraceus

    Directory of Open Access Journals (Sweden)

    Marilena Budroni

    2012-12-01

    Full Text Available The aim of this study was to select wine yeast strains as biocontrol agents against fungal contaminants responsible for the accumulation of ochratoxin A (OTA in grape and wine and to dissect the mechanism of OTA detoxification by a Saccharomyces cerevisiae strain (DISAABA1182, which had previously been reported to reduce OTA in a synthetic must. All of the yeast strains tested displayed an ability to inhibit the growth of Aspergillus carbonarius both in vivo and in vitro and addition of culture filtrates from the tested isolates led to complete inhibition of OTA production. S. cerevisiae DISAABA1182 was selected and further tested for its capacity to inhibit OTA production and pks (polyketide synthase transcription in A. carbonarius and Aspergillus ochraceus in vitro. In order to dissect the mechanism of OTA detoxification, each of these two fungi was co-cultured with living yeast cells exposed to yeast crude or to autoclaved supernatant: S. cerevisiae DISAABA1182 was found to inhibit mycelial growth and OTA production in both Aspergilli when co-cultured in the OTA-inducing YES medium. Moreover, a decrease in pks transcription was observed in the presence of living cells of S. cerevisiae DISAABA1182 or its supernatant, while no effects were observed on transcription of either of the constitutively expressed calmodulin and β-tubulin genes. This suggests that transcriptional regulation of OTA biosynthetic genes takes place during the interaction between DISAABA1182 and OTA-producing Aspergilli.

  15. The opposite roles of agdA and glaA on citric acid production in Aspergillus niger.

    Science.gov (United States)

    Wang, Lu; Cao, Zhanglei; Hou, Li; Yin, Liuhua; Wang, Dawei; Gao, Qiang; Wu, Zhenqiang; Wang, Depei

    2016-07-01

    Citric acid is produced by an industrial-scale process of fermentation using Aspergillus niger as a microbial cell factory. However, citric acid production was hindered by the non-fermentable isomaltose and insufficient saccharification ability in A. niger when liquefied corn starch was used as a raw material. In this study, A. niger TNA 101ΔagdA was constructed by deletion of the α-glucosidase-encoding agdA gene in A. niger CGMCC 10142 genome using Agrobacterium tumefaciens-mediated transformation. The transformants A. niger OG 1, OG 17, and OG 31 then underwent overexpression of glucoamylase in A. niger TNA 101ΔagdA. The results showed that the α-glucosidase activity of TNA 101ΔagdA was decreased by 62.5 % compared with CGMCC 10142, and isomaltose was almost undetectable in the fermentation broth. The glucoamylase activity of the transformants OG 1 and OG 17 increased by 34.5 and 16.89 % compared with that of TNA 101ΔagdA, respectively. In addition, for the recombinants TNA 101ΔagdA, OG 1 and OG 17, there were no apparent defects in the growth development. Consequently, in comparison with CGMCC 10142, TNA 101ΔagdA and OG 1 decreased the residual reducing sugar by 52.95 and 88.24 %, respectively, and correspondingly increased citric acid production at the end of fermentation by 8.68 and 16.87 %. Citric acid production was further improved by decreasing the non-fermentable residual sugar and increasing utilization rate of corn starch material in A. niger. Besides, the successive saccharification and citric acid fermentation processes were successfully integrated into one step.

  16. Insulin-like growth factor-I extends in vitro replicative life span of skeletal muscle satellite cells by enhancing G1/S cell cycle progression via the activation of phosphatidylinositol 3'-kinase/Akt signaling pathway

    Science.gov (United States)

    Chakravarthy, M. V.; Abraha, T. W.; Schwartz, R. J.; Fiorotto, M. L.; Booth, F. W.

    2000-01-01

    Interest is growing in methods to extend replicative life span of non-immortalized stem cells. Using the insulin-like growth factor I (IGF-I) transgenic mouse in which the IGF-I transgene is expressed during skeletal muscle development and maturation prior to isolation and during culture of satellite cells (the myogenic stem cells of mature skeletal muscle fibers) as a model system, we elucidated the underlying molecular mechanisms of IGF-I-mediated enhancement of proliferative potential of these cells. Satellite cells from IGF-I transgenic muscles achieved at least five additional population doublings above the maximum that was attained by wild type satellite cells. This IGF-I-induced increase in proliferative potential was mediated via activation of the phosphatidylinositol 3'-kinase/Akt pathway, independent of mitogen-activated protein kinase activity, facilitating G(1)/S cell cycle progression via a down-regulation of p27(Kip1). Adenovirally mediated ectopic overexpression of p27(Kip1) in exponentially growing IGF-I transgenic satellite cells reversed the increase in cyclin E-cdk2 kinase activity, pRb phosphorylation, and cyclin A protein abundance, thereby implicating an important role for p27(Kip1) in promoting satellite cell senescence. These observations provide a more complete dissection of molecular events by which increased local expression of a growth factor in mature skeletal muscle fibers extends replicative life span of primary stem cells than previously known.

  17. Two novel aflatoxin-producing Aspergillus species from Argentinean peanuts

    DEFF Research Database (Denmark)

    Pildain, M.B.; Frisvad, Jens Christian; Vaamonde, G.

    2008-01-01

    Two novel species from Aspergillus section Flavi from different species of Arachis (peanuts) in Argentina are described as Aspergillus arachidicola sp. nov. and Aspergillus minisclerotigenes sp. nov. Their novel taxonomic status was determined using a polyphasic taxonomic approach with phenotypic...... (morphology and extrolite profiles) and molecular (beta-tubulin and calmodulin gene sequences) characters. A. minisclerotigenes resembles Aspergillus flavus and Aspergillus parvisclerotigenus in producing aflatoxins B-1 and B-2, cyclopiazonic acid, kojic acid and aspergillic acid, but in addition it produces...... and parasiticolide, and some strains produce aspergillic acid. The type strain of A. arachidicola is CBS 117610(T) =IBT 25020(T) and that of A. minisclerotigenes is CBS 117635(T) =IBT 27196(T). The Mycobank accession numbers for Aspergillus minisclerotigenes sp. nov. and Aspergillus arachidicola sp. nov...

  18. Factorials of real negative and imaginary numbers - A new perspective

    OpenAIRE

    Thukral, Ashwani K

    2014-01-01

    Presently, factorials of real negative numbers and imaginary numbers, except for zero and negative integers are interpolated using the Euler’s gamma function. In the present paper, the concept of factorials has been generalised as applicable to real and imaginary numbers, and multifactorials. New functions based on Euler’s factorial function have been proposed for the factorials of real negative and imaginary numbers. As per the present concept, the factorials of real negative numbers, are co...

  19. Korean Ginseng Berry Fermented by Mycotoxin Non-producing Aspergillus niger and Aspergillus oryzae: Ginsenoside Analyses and Anti-proliferative Activities.

    Science.gov (United States)

    Li, Zhipeng; Ahn, Hyung Jin; Kim, Nam Yeon; Lee, Yu Na; Ji, Geun Eog

    2016-01-01

    To transform ginsenosides, Korean ginseng berry (KGB) was fermented by mycotoxin non-producing Aspergillus niger and Aspergillus oryzae. Changes of ginsenoside profile and anti-proliferative activities were observed. Results showed that A. niger tended to efficiently transform protopanaxadiol (PPD) type ginsenosides such as Rb1, Rb2, Rd to compound K while A. oryzae tended to efficiently transform protopanaxatriol (PPT) type ginsenoside Re to Rh1 via Rg1. Butanol extracts of fermented KGB showed high cytotoxicity on human adenocarcinoma HT-29 cell line and hepatocellular carcinoma HepG2 cell line while that of unfermented KGB showed little. The minimum effective concentration of niger-fermented KGB was less than 2.5 µg/mL while that of oryzae-fermented KGB was about 5 µg/mL. As A. niger is more inclined to transform PPD type ginsenosides, niger-fermented KGB showed stronger anti-proliferative activity than oryzae-fermented KGB.

  20. Taxonomic Characterization and Secondary Metabolite Profiling of Aspergillus Section Aspergillus Contaminating Feeds and Feedstuffs

    Science.gov (United States)

    Greco, Mariana; Kemppainen, Minna; Pose, Graciela; Pardo, Alejandro

    2015-01-01

    Xerophilic fungal species of the genus Aspergillus are economically highly relevant due to their ability to grow on low water activity substrates causing spoilage of stored goods and animal feeds. These fungi can synthesize a variety of secondary metabolites, many of which show animal toxicity, creating a health risk for food production animals and to humans as final consumers, respectively. Animal feeds used for rabbit, chinchilla and rainbow trout production in Argentina were analysed for the presence of xerophilic Aspergillus section Aspergillus species. High isolation frequencies (>60%) were detected in all the studied rabbit and chinchilla feeds, while the rainbow trout feeds showed lower fungal charge (25%). These section Aspergillus contaminations comprised predominantly five taxa. Twenty isolates were subjected to taxonomic characterization using both ascospore SEM micromorphology and two independent DNA loci sequencing. The secondary metabolite profiles of the isolates were determined qualitatively by HPLC-MS. All the isolates produced neoechinulin A, 17 isolates were positive for cladosporin and echinulin, and 18 were positive for neoechinulin B. Physcion and preechinulin were detected in a minor proportion of the isolates. This is the first report describing the detailed species composition and the secondary metabolite profiles of Aspergillus section Aspergillus contaminating animal feeds. PMID:26364643

  1. Taxonomic Characterization and Secondary Metabolite Profiling of Aspergillus Section Aspergillus Contaminating Feeds and Feedstuffs.

    Science.gov (United States)

    Greco, Mariana; Kemppainen, Minna; Pose, Graciela; Pardo, Alejandro

    2015-09-02

    Xerophilic fungal species of the genus Aspergillus are economically highly relevant due to their ability to grow on low water activity substrates causing spoilage of stored goods and animal feeds. These fungi can synthesize a variety of secondary metabolites, many of which show animal toxicity, creating a health risk for food production animals and to humans as final consumers, respectively. Animal feeds used for rabbit, chinchilla and rainbow trout production in Argentina were analysed for the presence of xerophilic Aspergillus section Aspergillus species. High isolation frequencies (>60%) were detected in all the studied rabbit and chinchilla feeds, while the rainbow trout feeds showed lower fungal charge (25%). These section Aspergillus contaminations comprised predominantly five taxa. Twenty isolates were subjected to taxonomic characterization using both ascospore SEM micromorphology and two independent DNA loci sequencing. The secondary metabolite profiles of the isolates were determined qualitatively by HPLC-MS. All the isolates produced neoechinulin A, 17 isolates were positive for cladosporin and echinulin, and 18 were positive for neoechinulin B. Physcion and preechinulin were detected in a minor proportion of the isolates. This is the first report describing the detailed species composition and the secondary metabolite profiles of Aspergillus section Aspergillus contaminating animal feeds.

  2. Taxonomic Characterization and Secondary Metabolite Profiling of Aspergillus Section Aspergillus Contaminating Feeds and Feedstuffs

    Directory of Open Access Journals (Sweden)

    Mariana Greco

    2015-09-01

    Full Text Available Xerophilic fungal species of the genus Aspergillus are economically highly relevant due to their ability to grow on low water activity substrates causing spoilage of stored goods and animal feeds. These fungi can synthesize a variety of secondary metabolites, many of which show animal toxicity, creating a health risk for food production animals and to humans as final consumers, respectively. Animal feeds used for rabbit, chinchilla and rainbow trout production in Argentina were analysed for the presence of xerophilic Aspergillus section Aspergillus species. High isolation frequencies (>60% were detected in all the studied rabbit and chinchilla feeds, while the rainbow trout feeds showed lower fungal charge (25%. These section Aspergillus contaminations comprised predominantly five taxa. Twenty isolates were subjected to taxonomic characterization using both ascospore SEM micromorphology and two independent DNA loci sequencing. The secondary metabolite profiles of the isolates were determined qualitatively by HPLC-MS. All the isolates produced neoechinulin A, 17 isolates were positive for cladosporin and echinulin, and 18 were positive for neoechinulin B. Physcion and preechinulin were detected in a minor proportion of the isolates. This is the first report describing the detailed species composition and the secondary metabolite profiles of Aspergillus section Aspergillus contaminating animal feeds.

  3. Mycotic keratitis due to Aspergillus nomius

    NARCIS (Netherlands)

    Manikandan, P.; Varga, J.; Kocsube, S.; Samson, R.A.; Anita, R.; Revathi, R.; Doczi, I.; Nemeth, T.M.; Narendran, V.; Vagvolgyi, C.; Manoharan, C.; Kredics, L.

    2009-01-01

    We report the first known case of fungal keratitis caused by Aspergillus nomius. Ocular injury was known as a predisposing factor. The patient was treated with natamycin and econazole eye drops, itraconazole eye ointment, and oral ketoconazole. A therapeutic penetrating keratoplasty was performed 16

  4. Aspergillus PCR: one step closer to standardization.

    NARCIS (Netherlands)

    White, P.L.; Bretagne, S.; Klingspor, L.; Melchers, W.J.G.; McCulloch, E.; Schulz, B.; Finnstrom, N.; Mengoli, C.; Barnes, R.A.; Donnelly, J.P.; Loeffler, J.

    2010-01-01

    PCR has been used as an aid in the diagnosis of invasive aspergillosis for almost 2 decades. A lack of standardization has limited both its acceptance as a diagnostic tool and multicenter clinical evaluations, preventing its inclusion in disease-defining criteria. In 2006, the European Aspergillus P

  5. Polyphasic taxonomy of Aspergillus section Cervini

    DEFF Research Database (Denmark)

    Chen, A.J.; Varga, J.; Frisvad, Jens Christian

    2016-01-01

    Species belonging to Aspergillus section Cervini are characterised by radiate or short columnar, fawn coloured, uniseriate conidial heads. The morphology of the taxa in this section is very similar and isolates assigned to these species are frequently misidentified. In this study, a polyphasic...

  6. Organic acid production by Aspergillus niger

    DEFF Research Database (Denmark)

    Jongh, Wian de

    2006-01-01

    . Specielt Aspergillus niger er interessant i forbindelse med produktion af organiske syrer, idet denne organisme tolerer lavt pH, kan give høje produktudbytter, og kan give høje produktiviteter som allerede illustreret i anvendelsen af denne organisme i produktionen af citronsyre. Disse faktorer gør A...

  7. Genetics of Polyketide Metabolism in Aspergillus nidulans

    DEFF Research Database (Denmark)

    Klejnstrup, Marie L.; Frandsen, Rasmus John Normand; Holm, Dorte Koefoed

    2012-01-01

    are publicly available, greatly facilitating the establishment of links between genes and metabolites. This review covers the attempts being made to trigger the activation of polyketide metabolism in the fungal model organism Aspergillus nidulans. Moreover, it will provide an overview of the pathways where ten...

  8. Interaction between maize seed and Aspergillus flavus

    Science.gov (United States)

    Aspergillus flavus is an opportunistic fungal pathogen that colonizes maize seeds and contaminates them with aflatoxin. The fungus is localized in the endosperm and aleurone. To investigate the plant microbe interaction, we conducted histological and molecular studies to characterize the internal co...

  9. Rapid Application Development Using Software Factories

    CERN Document Server

    Stojanovski, Toni

    2012-01-01

    Software development is still based on manufactory production, and most of the programming code is still hand-crafted. Software development is very far away from the ultimate goal of industrialization in software production, something which has been achieved long time ago in the other industries. The lack of software industrialization creates an inability to cope with fast and frequent changes in user requirements, and causes cost and time inefficiencies during their implementation. Analogous to what other industries had done long time ago, industrialization of software development has been proposed using the concept of software factories. We have accepted this vision about software factories, and developed our own software factory which produces three-layered ASP.NET web applications. In this paper we report about our experience with using this approach in the process of software development, and present comparative results on performances and deliverables in both traditional development and development usin...

  10. Low-energy neutrino factory design

    Energy Technology Data Exchange (ETDEWEB)

    Ankenbrandt, C.; /Fermilab /MUONS Inc., Batavia; Bogacz, S.A.; /Jefferson Lab; Bross, A.; Geer, S.; Johnstone, C.; Neuffer, D.; Popovic, M.; /Fermilab

    2009-07-01

    The design of a low-energy (4 GeV) neutrino factory (NF) is described, along with its expected performance. The neutrino factory uses a high-energy proton beam to produce charged pions. The {pi}{sup {+-}} decay to produce muons ({mu}{sup {+-}}), which are collected, accelerated, and stored in a ring with long straight sections. Muons decaying in the straight sections produce neutrino beams. The scheme is based on previous designs for higher energy neutrino factories, but has an improved bunching and phase rotation system, and new acceleration, storage ring, and detector schemes tailored to the needs of the lower energy facility. Our simulations suggest that the NF scheme we describe can produce neutrino beams generated by {approx} 1.4 x 10{sup 21} {mu}{sup +} per year decaying in a long straight section of the storage ring, and a similar number of {mu}{sup -} decays.

  11. Ergonomic analysis jobs in recovered factories.

    Science.gov (United States)

    Cuenca, Gabriela; Zotta, Gastón

    2012-01-01

    With the advent of the deep economic crisis in Argentina on 2001, the recovery of companies through to the creation of the Cooperatives Working Self-Management or Factories Recovered by its workers was constituted as one of the ways in which the salaried disobeyed the increasing unemployment. When the companies turn into recovered factories they tend to leave of side practices that have been seen like imposed by the previous organization and not understanding them as a primary condition for the execution of his tasks. Safety and ergonomics are two disciplines that are no longer considered relevant to the daily work. Therefore this investigation aims to revalue, undergo semantic to give back to a place in every organization analyzed. This research developed a self-diagnostic tool for working conditions, and the environment, present in the recovered factories.

  12. Ageratum conyzoides essential oil as aflatoxin suppressor of Aspergillus flavus.

    Science.gov (United States)

    Nogueira, Juliana H C; Gonçalez, Edlayne; Galleti, Silvia R; Facanali, Roseane; Marques, Márcia O M; Felício, Joana D

    2010-01-31

    Aflatoxin B(1) (AFB(1)) is a highly toxic and carcinogenic metabolite produced by Aspergillus species on food and agricultural commodities. Inhibitory effects of essential oil of Ageratum conyzoides, on the mycelial growth and aflatoxin B(1) production by Aspergillus flavus were studied. Cultures were incubated in yeast extract-sucrose (YES) broth for days at 25 degrees C at the following different concentrations of the essential oil (from 0.0 to 30mug/mL). The essential oil inhibited fungal growth to different extents depending on the concentration, and completely inhibited aflatoxin production at concentrations above 0.10microg/mL. The analysis of the oil by GC/MS showed that its main components are precocene II (46.35%), precocene I (42.78%), cumarine (5.01%) and Trans-caryophyllene (3.02%). Comparison by transmission electron microscopy of the fungal cells, control and those incubated with different concentrations of essential oil, showed ultra-structural changes which were concentration dependent of the essential oil of A. conyzoides. Such ultra-structural changes were more evident in the endomembrane system, affecting mainly the mitochondria. Degradation was also observed in both surrounding fibrils. The ability to inhibit aflatoxin production as a new biological activity of A.conyzoides L. indicates that it may be considered as a useful tool for a better understanding of the complex pathway of aflatoxin biosynthesis.

  13. Cadmium biosorption by Aspergillus niger; Biossorcao de cadmio pelo Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Silva, E.P.; Barros Junior, L.M.; Duarte, M.M.L.; Macedo, G.R. [Universidade Federal do Rio Grande do Norte, Natal, RN (Brazil)]. E-mail: edmilson@eq.ufrn.br

    2003-07-01

    Biosorption is a property of certain types of inactive, dead, microbial biomass to bind and concentrate heavy metals from even very dilute aqueous solutions. Biomass exhibits this property, acting just as a chemical substance, as an ion exchanger of biological origin. It is particularly the cell wall structure of certain algae, fungi and bacteria which was found responsible for this phenomenon. Some of the biomass types come as a waste by-product of large-scale industrial fermentations (the mold Rhizopus or the bacterium Bacillus subtilis). Other metal-binding biomass types, certain abundant seaweeds (particularly brown algae e.g. Sargassum, Ecklonia), can be readily collected from the oceans. These biomass types, serving as a basis for metal biosorption processes, can accumulate in excess of 25% of their dry weight in deposited heavy metals: Pb, Cd, U, Cu, Zn, even Cr and others. Sorption experiments using the Aspergillus niger fungus for cadmium removal were carried out to study the factors influencing and optimizing the biosorption of this metal. The effects of pH, time, biomass concentration, and initial concentration of the heavy metal on the rate of metallic biosorption were examined. (author)

  14. Influence of tallow and Aspergillus oryzae fermentation extract in dairy cattle rations.

    Science.gov (United States)

    Bertrand, J A; Grimes, L W

    1997-06-01

    Objectives were to determine the effects of adding 3 g/d of Aspergillus oryzae fermentation extract to diets with or without 5.6% added tallow. Twenty-eight Holstein cows (mean = 98 d of lactation) were assigned to a randomized block experiment in a 2 x 2 factorial arrangement of treatments. Treatments were the basal diet 1) without tallow or extract, 2) with extract but no tallow, 3) with tallow but no extract, and 4) with tallow and extract. Milk production, dry matter intake, 3.5% fat corrected milk, digestibility of neutral detergent fiber in the total tract were depressed for cows fed tallow. Addition of fermentation extract did not stimulate fiber digestion or milk production of cows fed diets with or without fat. Addition of extract did not overcome depression of fiber digestibility by cows fed tallow.

  15. Remembering the Avonmouth mustard gas factory

    OpenAIRE

    2014-01-01

    Introduction \\ud \\ud The women of the Great War had undertaken a predominantly domestic role until that point in time. This changed during these testing times, as women took the place of the men in the factories. There was an urgent need to produce large quantities of ammunition and the women made up the majority of the workforce. In the factories of Avonmouth and Chittening hundreds of women worked in perilous conditions to produce shells, but these shells weren’t just filled with explosives...

  16. Aspergillus niger contains the cryptic phylogenetic species A. awamori.

    Science.gov (United States)

    Perrone, Giancarlo; Stea, Gaetano; Epifani, Filomena; Varga, János; Frisvad, Jens C; Samson, Robert A

    2011-11-01

    Aspergillus section Nigri is an important group of species for food and medical mycology, and biotechnology. The Aspergillus niger 'aggregate' represents its most complicated taxonomic subgroup containing eight morphologically indistinguishable taxa: A. niger, Aspergillus tubingensis, Aspergillus acidus, Aspergillus brasiliensis, Aspergillus costaricaensis, Aspergillus lacticoffeatus, Aspergillus piperis, and Aspergillus vadensis. Aspergillus awamori, first described by Nakazawa, has been compared taxonomically with other black aspergilli and recently it has been treated as a synonym of A. niger. Phylogenetic analyses of sequences generated from portions of three genes coding for the proteins β-tubulin (benA), calmodulin (CaM), and the translation elongation factor-1 alpha (TEF-1α) of a population of A. niger strains isolated from grapes in Europe revealed the presence of a cryptic phylogenetic species within this population, A. awamori. Morphological, physiological, ecological and chemical data overlap occurred between A. niger and the cryptic A. awamori, however the splitting of these two species was also supported by AFLP analysis of the full genome. Isolates in both phylospecies can produce the mycotoxins ochratoxin A and fumonisin B₂, and they also share the production of pyranonigrin A, tensidol B, funalenone, malformins, and naphtho-γ-pyrones. In addition, sequence analysis of four putative A. awamori strains from Japan, used in the koji industrial fermentation, revealed that none of these strains belong to the A. awamori phylospecies.

  17. New taxa in Aspergillus section Usti.

    Science.gov (United States)

    Samson, R A; Varga, J; Meijer, M; Frisvad, J C

    2011-06-30

    Based on phylogenetic analysis of sequence data, Aspergillus section Usti includes 21 species, inclucing two teleomorphic species Aspergillus heterothallicus (= Emericella heterothallica) and Fennellia monodii. Aspergillus germanicus sp. nov. was isolated from indoor air in Germany. This species has identical ITS sequences with A. insuetusCBS 119.27, but is clearly distinct from that species based on β-tubulin and calmodulin sequence data. This species is unable to grow at 37 °C, similarly to A. keveii and A. insuetus. Aspergillus carlsbadensis sp. nov. was isolated from the Carlsbad Caverns National Park in New Mexico. This taxon is related to, but distinct from a clade including A. calidoustus, A. pseudodeflectus, A. insuetus and A. keveii on all trees. This species is also unable to grow at 37 °C, and acid production was not observed on CREA. Aspergillus californicus sp. nov. is proposed for an isolate from chamise chaparral (Adenostoma fasciculatum) in California. It is related to a clade including A. subsessilis and A. kassunensis on all trees. This species grew well at 37 °C, and acid production was not observed on CREA. The strain CBS 504.65 from soil in Turkey showed to be clearly distinct from the A. deflectus ex-type strain, indicating that this isolate represents a distinct species in this section. We propose the name A. turkensis sp. nov. for this taxon. This species grew, although rather restrictedly at 37 °C, and acid production was not observed on CREA. Isolates from stored maize, South Africa, as a culture contaminant of Bipolaris sorokiniana from indoor air in Finland proved to be related to, but different from A. ustus and A. puniceus. The taxon is proposed as the new species A. pseudoustus. Although supported only by low bootstrap values, F. monodii was found to belong to section Usti based on phylogenetic analysis of either loci BLAST searches to the GenBank database also resulted in closest hits from section Usti. This species obviously

  18. Software Development Factories, the Project Management Perspective

    OpenAIRE

    Felician ALECU

    2012-01-01

    In a software factory, the applications are developed in the same way Henri Ford started manufacturing cars. In such a way, a customized solution could be obtained within a reasonable budget and within the foreseen timeframe. This development method offers a flexible solution that can be readapted rapidly and automatically to the changing business needs, so the Project Management becomes a real challenge.

  19. The Energy Factory; EnergieFabriek

    Energy Technology Data Exchange (ETDEWEB)

    Van den Boomen, M.; Van den Dungen, G.J.; Elias, T.; Jansen, M. [Universiteit van Amsterdam UvA, Amsterdam (Netherlands)

    2009-05-15

    The Energy Factory is a collaboration of 26 Dutch local water boards in which options for energy saving and energy production are examined. According to the authors, the initiative of the Energy Factory will lead to a reframing of the role of the water boards. Moreover, they explain how the PPP concept (People, Planet, Profit) may act as platform for negotiations between actors who are involved in the Energy Factory. In addition, the PPP concept is used to demonstrate that the Energy Factory will lead to larger social involvement, social entrepreneurship and growing profits [Dutch] De Energiefabriek is een samenwerkingsverband van 26 waterschappen in Nederland waarin wordt gezocht naar mogelijkheden om energie te besparen en zelf energie te produceren. Volgens de auteurs van deze notitie leidt het initiatief van de Energiefabriek tot een reframing van de rol van waterschappen. Daarnaast leggen ze uit hoe het PPP-concept (People, Profit, Planet) kan fungeren als platform voor onderhandelingen tussen de actoren die betrokken zijn bij de Energiefabriek. Verder wordt met het PPP-concept aangetoond dat de Energiefabriek leidt tot ruimere maatschappelijke betrokkenheid, maatschappelijk ondernemen en winstvergroting.

  20. Business plan Feed Factory Zambezi Valley, Mozambique

    NARCIS (Netherlands)

    Vernooij, A.G.; Wilschut, S.

    2015-01-01

    This business plan focuses on the establishment of a poultry feed factory, one of the essential elements of a sustainable and profitable poultry meat value chain. There is a growing demand for poultry meat in the Tete Province, and currently a large part of the consumed broilers come from other part

  1. Guided and Interactive Factory Tours for Schools

    NARCIS (Netherlands)

    Kaibel, Andreas; Auwaerter, Andreas; Kravcik, Milos

    2006-01-01

    Please, cite this paper as: Kaibel, A., Auwaerter, A., & Kravcik, M. (2006). Guided and Interactive Factory Tours for Schools. Proceedings of the First European Conference on Technology Enhanced Learning. October 1st-4th, Crete, Greece: Springer. Retrieved October 18th, 2006, from http://dspace.lear

  2. The Idea Factory: An Interactive Intergroup Exercise

    Science.gov (United States)

    Rosh, Lisa; Leach, Evan

    2011-01-01

    This article outlines the Idea Factory exercise, an interactive exercise designed to help participants examine group, individual, and organizational factors that affect intergroup conflict. Specific emphasis is placed on exploring the relationship between intra- and intergroup dynamics and identifying managerial practices that foster effective…

  3. An Epiphany in a Toilet Factory

    Science.gov (United States)

    Cobb, Catlin

    2009-01-01

    In this article, the author shares her experiences of the universality of art making and artistic experience of being a dancer and teaching artist. She describes her performance at Kohler, a worldwide leader in plumbing products in Spartanburg, South Carolina, where she had an epiphany dancing in a toilet factory--a sudden, intuitive moment of…

  4. Jean Desmet’s Dream Factory

    Directory of Open Access Journals (Sweden)

    Eline Grignard

    2015-07-01

    Full Text Available Exhibition review of "Jean Desmet’s Dream Factory. The Adventurous Years of Film (1907-1916" held in Eye Film Instituut in Amsterdam. Paying hommage to film operator and collector Jean Desmet, the exhibition presents a wide range of the film collection as well as a number of rare archival materials.

  5. Roles of factorial noise in inducing bimodal gene expression

    Science.gov (United States)

    Liu, Peijiang; Yuan, Zhanjiang; Huang, Lifang; Zhou, Tianshou

    2015-06-01

    Some gene regulatory systems can exhibit bimodal distributions of mRNA or protein although the deterministic counterparts are monostable. This noise-induced bimodality is an interesting phenomenon and has important biological implications, but it is unclear how different sources of expression noise (each source creates so-called factorial noise that is defined as a component of the total noise) contribute separately to this stochastic bimodality. Here we consider a minimal model of gene regulation, which is monostable in the deterministic case. Although simple, this system contains factorial noise of two main kinds: promoter noise due to switching between gene states and transcriptional (or translational) noise due to synthesis and degradation of mRNA (or protein). To better trace the roles of factorial noise in inducing bimodality, we also analyze two limit models, continuous and adiabatic approximations, apart from the exact model. We show that in the case of slow gene switching, the continuous model where only promoter noise is considered can exhibit bimodality; in the case of fast switching, the adiabatic model where only transcriptional or translational noise is considered can also exhibit bimodality but the exact model cannot; and in other cases, both promoter noise and transcriptional or translational noise can cooperatively induce bimodality. Since slow gene switching and large protein copy numbers are characteristics of eukaryotic cells, whereas fast gene switching and small protein copy numbers are characteristics of prokaryotic cells, we infer that eukaryotic stochastic bimodality is induced mainly by promoter noise, whereas prokaryotic stochastic bimodality is induced primarily by transcriptional or translational noise.

  6. 21 CFR 173.120 - Carbohydrase and cellulase derived from Aspergillus niger.

    Science.gov (United States)

    2010-04-01

    ... cellulase derived from Aspergillus niger. Carbohydrase and cellulase enzyme preparation derived from Aspergillus niger may be safely used in food in accordance with the following prescribed conditions: (a) Aspergillus niger is classified as follows: Class, Deuteromycetes; order, Moniliales; family,...

  7. Two novel species of Aspergillus section Nigri from Thai coffee beans

    DEFF Research Database (Denmark)

    Noonim, Paramee; Mahakarnchanakul, Warapa; Varga, Janos

    2008-01-01

    Two novel species of Aspergillus section Nigri from Thai coffee beans are described as Aspergillus aculeatinus sp. nov. and Aspergillus sclerotiicarbonarius sp. nov. Their taxonomic status was determined using a polyphasic taxonomic approach with phenotypic (morphology and extrolite profiles...

  8. Phylogeny, identification and nomenclature of the genus Aspergillus

    DEFF Research Database (Denmark)

    Samson, R.A.; Visagie, C.M.; Houbraken, J.;

    2014-01-01

    Aspergillus comprises a diverse group of species based on morphological, physiological and phylogenetic characters, which significantly impact biotechnology, food production, indoor environments and human health. Aspergillus was traditionally associated with nine teleomorph genera, but phylogenetic...... data suggest that together with genera such as Polypaecilum, Phialosimplex, Dichotomomyces and Cristaspora, Aspergillus forms a monophyletic clade closely related to Penicillium. Changes in the International Code of Nomenclature for algae, fungi and plants resulted in the move to one name per species......, meaning that a decision had to be made whether to keep Aspergillus as one big genus or to split it into several smaller genera. The International Commission of Penicillium and Aspergillus decided to keep Aspergillus instead of using smaller genera. In this paper, we present the arguments for this decision...

  9. New and revisited species in Aspergillus section Nigri

    DEFF Research Database (Denmark)

    Varga, J.; Frisvad, Jens Christian; Kocsube, S.

    2011-01-01

    Four new species, Aspergillus eucalypticola, A. neoniger, A. fijiensis and A. indolo genus are described and illustrated. Aspergillus eucalypticola was isolated from Eucalyptus leaf from Australia, and is related to A. tubingensis and A. costaricaensis, but could clearly be distinguished from them...... based on either beta-tubulin or calmodulin sequence data. Aspergillus eucalypticola produced pyranonigrin A, funalenone, aurasperone B and other naphtho-gamma-pyrones. Aspergillus neoniger is also a biseriate species isolated from desert sand in Namibia, and mangrove water in Venezuela, which produces...... aurasperone B and pyranonigrin A. Aspergillus fijiensis is a uniseriate species related to A. aculeatinus, and was isolated from soil in Fiji, and from Lactuca saliva in Indonesia. This species is able to grow at 37 degrees C, and produces asperparalines and okaramins. Aspergillus indologenus was isolated...

  10. Aflatoxigenic Aspergillus flavus and Aspergillus parasiticus strains in Hungarian maize fields.

    Science.gov (United States)

    Sebők, Flóra; Dobolyi, Csaba; Zágoni, Dóra; Risa, Anita; Krifaton, Csilla; Hartman, Mátyás; Cserháti, Mátyás; Szoboszlay, Sándor; Kriszt, Balázs

    2016-12-01

    Due to the climate change, aflatoxigenic Aspergillus species and strains have appeared in several European countries, contaminating different agricultural commodities with aflatoxin. Our aim was to screen the presence of aflatoxigenic fungi in maize fields throughout the seven geographic regions of Hungary. Fungi belonging to Aspergillus section Flavi were isolated in the ratio of 26.9% and 42.3% from soil and maize samples in 2013, and these ratios decreased to 16.1% and 34.7% in 2014. Based on morphological characteristics and the sequence analysis of the partial calmodulin gene, all isolates proved to be Aspergillus flavus, except four strains, which were identified as Aspergillus parasiticus. About half of the A. flavus strains and all the A. parasiticus strains were able to synthesize aflatoxins. Aflatoxigenic Aspergillus strains were isolated from all the seven regions of Hungary. A. parasiticus strains were found in the soil of the regions Southern Great Plain and Southern Transdanubia and in a maize sample of the region Western Transdanubia. In spite of the fact that aflatoxins have rarely been detected in feeds and foods in Hungary, aflatoxigenic A. flavus and A. parasiticus strains are present in the maize culture throughout Hungary posing a potential threat to food safety.

  11. Optimization of Cassava Waste from Bioethanol Post-Production through Bioactivity Process Consortium of Saccharomyces cerevisiae, Trichoderma viride and Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Yani Suryani

    2013-03-01

    Full Text Available The result of laboratory analysis on bioethanol waste shows nutritional and anti-nutritional content (HCN. Based on it, in order to prevent environment damage then performed a bioconversion of bioethanol waste for base material of sheep forage by fermented previously using consortium of Saccharomyces cerevisiae, Trichoderma viride, and Aspergillus niger to eliminate the waste anti-nutritional content and increase nutritional content. This research used Completely Randomized Design (CRD factorial pattern with two factors and three replications. The first factor was inoculum dose (D and the second factor was variation of tested microorganisms (M. The variables measured were HCN content and nutrient content of fermented products, that were water content, protein content and crude fiber content by proximate analysis. Furthermore, the data was statistically analyzed by analysis of variance (ANOVA, and was there significant difference analyzed further by Duncan's multiple range test of 5% level. Nutritional compositions of bioethanol waste from cassava that had been fermented by consortium of Saccharomyces cerevisiae, Trichoderma viride, and Aspergillus niger experienced changes compared to before fermentation. The results showed an increase in protein level, while fiber content, water content, and HCN content were decrease. Bioethanol waste from cassava fermented by Aspergillus niger and Saccharomyces cerevisiae (k3 with 2% inoculum dose had the highest protein content increase from 11.79% to 25.41% and had the lowest crude fiber content decrease from 16.4% to 12.84 %. The highest HCN decrease obtained from the fermentation of consortium Aspergillus niger and Trichoderma viride.

  12. Immunological Aspects of Candida and Aspergillus Systemic Fungal Infections

    Directory of Open Access Journals (Sweden)

    Christoph Mueller-Loebnitz

    2013-01-01

    Full Text Available Patients with allogeneic stem cell transplantation (SCT have a high risk of invasive fungal infections (IFIs even after neutrophil regeneration. Immunological aspects might play a very important role in the IFI development in these patients. Some data are available supporting the identification of high-risk patients with IFI for example patients receiving stem cells from TLR4 haplotype S4 positive donors. Key defense mechanisms against IFI include the activation of neutrophils, the phagocytosis of germinating conidia by dendritic cells, and the fight of the cells of the innate immunity such as monocytes and natural killer cells against germlings and hyphae. Furthermore, immunosuppressive drugs interact with immune effector cells influencing the specific fungal immune defense and antimycotic drugs might interact with immune response. Based on the current knowledge on immunological mechanism in Aspergillus fumigatus, the first approaches of an immunotherapy using human T cells are in development. This might be an option for the future of aspergillosis patients having a poor prognosis with conventional treatment.

  13. In vitro interactions of antifungal agents and tacrolimus against Aspergillus biofilms.

    Science.gov (United States)

    Gao, Lujuan; Sun, Yi

    2015-11-01

    Aspergillus biofilms were prepared from Aspergillus fumigatus, Aspergillus flavus, and Aspergillus terreus via a 96-well plate-based method, and the combined antifungal activity of tacrolimus with azoles or amphotericin B against Aspergillus biofilms was investigated via a broth microdilution checkerboard technique system. Our results suggest that combinations of tacrolimus with voriconazole or amphotericin B have synergistic inhibitory activity against Aspergillus biofilms. However, combinations of tacrolimus with itraconazole or posaconazole exhibit no synergistic or antagonistic effects.

  14. Aspergillus deflectus infection in four dogs.

    Science.gov (United States)

    Jang, S S; Dorr, T E; Biberstein, E L; Wong, A

    1986-04-01

    Four cases of disseminated aspergillosis caused by Aspergillus deflectus in German Shepherds are presented. Three of the cases, which involved multiple organs, terminated in euthanasia. One case, with bony involvement of the limbs and skull, lived. The unique morphological characteristic of the conidial head resembling a briar pipe led to the identification of A. deflectus. To the authors' knowledge these are the first reported cases of infections caused by A. deflectus in man or animal.

  15. Production of Tetraquark State Tcc at B-Factories

    Science.gov (United States)

    Reyima, Rashidin

    2013-12-01

    We study production of the tetraquark state Tcc via virtual photon at the B-factories in the QCD factorization framework. We predict the cross section of tetraquark state production in the leading order at the B-factories.

  16. Aspergillus species: An emerging pathogen in onychomycosis among diabetics

    Directory of Open Access Journals (Sweden)

    T M Wijesuriya

    2015-01-01

    Full Text Available Introduction: Approximately, 33% patients with diabetes are afflicted with onychomycosis. In the past, nondermatophyte molds have been regarded as opportunistic pathogens; recently, Aspergillus species are considered as emerging pathogens of toenail infections. In Sri Lanka, the prevalence of Aspergillus species in onychomycosis among diabetics is not well documented. Objective: To determine the proportion of Aspergillus onychomycosis, risk factors and knowledge among diabetics. Materials and Methods: This was descriptive cross-sectional study. Three hundred diabetic patients were included. Clinical examinations of patients' toenails were performed by a clinical microbiologist. Laboratory identification was done, and pathogens were identified to the species level by morpho-physiological methods. All inferential statistics were tested at P age group. In men, Aspergillus onycomycosis was seen in 82%. Among patients who had Aspergillus nail infection, 114 (63% had diabetes for a period of > years. Among patients who were engaged in agricultural activities, 77% were confirmed to have infected nails due to Aspergillus species. Conclusion: Aspergillus niger was the most common pathogen isolated from toenail infection. Aspergillus species should be considered as an important pathogen in toenail onychomycosis in diabetic patients. Risk factors associated with Aspergillus onychomycosis were age, gender, duration of diabetes, length of exposure to fungi, and occupation.

  17. Aspergillus thyroiditis in a renal transplant recipient mimicking subacute thyroiditis.

    Science.gov (United States)

    Solak, Y; Atalay, H; Nar, A; Ozbek, O; Turkmen, K; Erekul, S; Turk, S

    2011-04-01

    Fungal pathogens are increasingly encountered after renal transplantation. Aspergillus causes significant morbidity and mortality in transplant patients. Fungal thyroiditis is a rare occurrence owing to unique features of the thyroid gland. Most cases are caused by Aspergillus species and have been described in immunocompromised patients. Presentation may be identical with that of subacute thyroiditis, in which hyperthyroidism features and painful thyroid are the prominent findings. Diagnosis can be ascertained by fine-needle aspiration of thyroid showing branching hyphae of Aspergillus. We describe a renal transplant patient who developed Aspergillus thyroiditis as part of a disseminated infection successfully treated with voriconazole.

  18. Protease-dependent activation of epithelial cells by fungal allergens leads to morphologic changes and cytokine production

    NARCIS (Netherlands)

    Kauffman, HF; Tomee, JFC; van de Riet, MA; Timmerman, AJB; Borger, P

    2000-01-01

    Background: Proteases in extracts of Aspergillus fumigatus cause epithelial cell desquamation and release of proinflammatory cytokines. Objective: We sought to assess protease activity in Alternaria alternata, Cladosporium herbarum, and Aspergillus fumigatus extracts and study the ability of these e

  19. Purification and properties of beta-galactosidase from Aspergillus nidulans.

    Science.gov (United States)

    Díaz, M; Pedregosa, A M; de Lucas, J R; Torralba, S; Monistrol, I F; Laborda, F

    1996-12-01

    Beta-Galactosidase from mycelial extract of Aspergillus nidulans has been purified by substrate affinity chromatography and used to obtain anti-beta-galactosidase polyclonal antibodies. A. nidulans growing in lactose as carbon source synthesizes one active form of beta-galactosidase which seems to be a multimeric enzyme of 450 kDa composed of monomers with 120 and 97 kDa. Although the enzyme was not released to the culture medium, some enzymatic activity was detected in a cell-wall extract, thus suggesting that it can be an extracellular enzyme. Beta-Galactosidase of A. nidulans is a very unstable enzyme with an optimum pH value of 7.5 and an optimum temperature of 30 degrees C. It was only active against beta-galactoside substrates like lactose and p-nitrophenyl-beta-D-galactoside (PNPG).

  20. Flocculation behavior and mechanism of bioflocculant produced by Aspergillus flavus.

    Science.gov (United States)

    Aljuboori, Ahmad H Rajab; Idris, Azni; Al-joubory, Hamid Hussain Rijab; Uemura, Yoshimitsu; Ibn Abubakar, B S U

    2015-03-01

    In this study, the flocculation behavior and mechanism of a cation-independent bioflocculant IH-7 produced by Aspergillus flavus were investigated. Results showed 91.6% was the lowest flocculating rate recorded by IH-7 (0.5 mg L(-1)) at pH range 4-8. Moreover, IH-7 showed better flocculation performance than polyaluminum chloride (PAC) at a wide range of flocculant concentration (0.06-25 mg L(-1)), temperature (5-45 °C) and salinity (10-60% w/w). The current study found that cation addition did not significantly enhance the flocculating rate and IH-7 is a positively charged bioflocculant. These findings suggest that charge neutralization is the main flocculation mechanism of IH-7 bioflocculant. IH-7 was significantly used to flocculate different types of suspended solids such as activated carbons, kaolin clays, soil solids and yeast cells.

  1. A new diketopiperazine heterodimer from an endophytic fungus Aspergillus niger.

    Science.gov (United States)

    Li, Xiao-Bin; Li, Yue-Lan; Zhou, Jin-Chuan; Yuan, Hui-Qing; Wang, Xiao-Ning; Lou, Hong-Xiang

    2015-01-01

    One new diketopiperazine heterodimer, asperazine A (1), and eight known compounds, asperazine (2), cyclo(d-Phe-l-Trp) (3), cyclo(l-Trp-l-Trp) (4), 4-(hydroxymethyl)-5,6-dihydro-pyran-2-one (5), walterolactone A (6), and campyrones A-C (7-9), were isolated from an endophytic fungus Aspergillus niger. Their structures were determined unequivocally on the basis of extensive spectroscopic data analysis. This is the first report of the presence of compound 3 as a natural product. Cytotoxicity test against human cancer cell lines PC3, A2780, K562, MBA-MD-231, and NCI-H1688 revealed that compounds 1 and 2 had weak activities.

  2. Biosolubilization of poorly soluble rock phosphates by Aspergillus tubingensis and Aspergillus niger.

    Science.gov (United States)

    Reddy, M Sudhakara; Kumar, Surender; Babita, K; Reddy, M S

    2002-09-01

    Three isolates of Aspergillus tubingensis and two isolates of Aspergillus niger isolated from rhizospheric soils were tested on solubilization of different rock phosphates. All the isolates of Aspergillus were capable of solubilizing all the natural rock phosphates. A. tubingensis (AT1) showed maximum percent solubilization in all the rock phosphates tested in this study when compared to other isolates. This isolate also showed highest phosphorus (P) solubilization when grown in the presence of 2% of rock phosphate. A. tubingensis (AT1) seems to be more efficient in solubilization of rock phosphates compared to other isolates reported elsewhere. This is the first report of rock phosphate solubilization by A. tubingensis and might provide an efficient large scale biosolubilization of rock phosphates intended for P fertilizer.

  3. Comparative Reannotation of 21 Aspergillus Genomes

    Energy Technology Data Exchange (ETDEWEB)

    Salamov, Asaf; Riley, Robert; Kuo, Alan; Grigoriev, Igor

    2013-03-08

    We used comparative gene modeling to reannotate 21 Aspergillus genomes. Initial automatic annotation of individual genomes may contain some errors of different nature, e.g. missing genes, incorrect exon-intron structures, 'chimeras', which fuse 2 or more real genes or alternatively splitting some real genes into 2 or more models. The main premise behind the comparative modeling approach is that for closely related genomes most orthologous families have the same conserved gene structure. The algorithm maps all gene models predicted in each individual Aspergillus genome to the other genomes and, for each locus, selects from potentially many competing models, the one which most closely resembles the orthologous genes from other genomes. This procedure is iterated until no further change in gene models is observed. For Aspergillus genomes we predicted in total 4503 new gene models ( ~;;2percent per genome), supported by comparative analysis, additionally correcting ~;;18percent of old gene models. This resulted in a total of 4065 more genes with annotated PFAM domains (~;;3percent increase per genome). Analysis of a few genomes with EST/transcriptomics data shows that the new annotation sets also have a higher number of EST-supported splice sites at exon-intron boundaries.

  4. Flow-based approach for holistic factory engineering and design

    OpenAIRE

    Constantinescu, C.; Westkämper, E.

    2010-01-01

    The engineering of future factories requires digital tools along life cycle phases from investment planning to ramp-up. Manufacturers need scientific-based integrated highly dynamic data management systems for the participative and integrated factory planning. The paper presents a new approach for the continuously integrated product design, factory and process planning, through a service-oriented architecture for the implementation of digital factory tools. A first prototype of the digital fa...

  5. An Interactive Simulation Tool for Production Planning in Bacon Factories

    DEFF Research Database (Denmark)

    Nielsen, Jens Frederik Dalsgaard; Nielsen, Kirsten Mølgaard

    1994-01-01

    The paper describes an interactive simulation tool for production planning in bacon factories. The main aim of the tool is to make it possible to combine the production plans of all parts of the factory......The paper describes an interactive simulation tool for production planning in bacon factories. The main aim of the tool is to make it possible to combine the production plans of all parts of the factory...

  6. Aspergillus nidulans galactofuranose biosynthesis affects antifungal drug sensitivity.

    Science.gov (United States)

    Alam, Md Kausar; El-Ganiny, Amira M; Afroz, Sharmin; Sanders, David A R; Liu, Juxin; Kaminskyj, Susan G W

    2012-12-01

    The cell wall is essential for fungal survival in natural environments. Many fungal wall carbohydrates are absent from humans, so they are a promising source of antifungal drug targets. Galactofuranose (Galf) is a sugar that decorates certain carbohydrates and lipids. It comprises about 5% of the Aspergillus fumigatus cell wall, and may play a role in systemic aspergillosis. We are studying Aspergillus wall formation in the tractable model system, A. nidulans. Previously we showed single-gene deletions of three sequential A. nidulans Galf biosynthesis proteins each caused similar hyphal morphogenesis defects and 500-fold reduced colony growth and sporulation. Here, we generated ugeA, ugmA and ugtA strains controlled by the alcA(p) or niiA(p) regulatable promoters. For repression and expression, alcA(p)-regulated strains were grown on complete medium with glucose or threonine, whereas niiA(p)-regulated strains were grown on minimal medium with ammonium or nitrate. Expression was assessed by qPCR and colony phenotype. The alcA(p) and niiA(p) strains produced similar effects: colonies resembling wild type for gene expression, and resembling deletion strains for gene repression. Galf immunolocalization using the L10 monoclonal antibody showed that ugmA deletion and repression phenotypes correlated with loss of hyphal wall Galf. None of the gene manipulations affected itraconazole sensitivity, as expected. Deletion of any of ugmA, ugeA, ugtA, their repression by alcA(p) or niiA(p), OR, ugmA overexpression by alcA(p), increased sensitivity to Caspofungin. Strains with alcA(p)-mediated overexpression of ugeA and ugtA had lower caspofungin sensitivity. Galf appears to play an important role in A. nidulans growth and vigor.

  7. Internet factories: Creating application-specific networks on-demand

    NARCIS (Netherlands)

    Strijkers, R.J.; Makkes, M.X.; Laat, C. de; Meijer, R.J.

    2014-01-01

    We introduce the concept of Internet factories. Internet factories structure the task of creating and managing application-specific overlay networks using infrastructure-as-a-service clouds. We describe the Internet factory architecture and report on a proof of concept with three examples that progr

  8. 27 CFR 40.47 - Other businesses within factory.

    Science.gov (United States)

    2010-04-01

    ... factory. 40.47 Section 40.47 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE... AND TUBES, AND PROCESSED TOBACCO Administrative Provisions § 40.47 Other businesses within factory. (a) General. The appropriate TTB officer may authorize such other businesses within the factory of...

  9. 46 CFR 164.009-23 - Factory inspection.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 6 2010-10-01 2010-10-01 false Factory inspection. 164.009-23 Section 164.009-23...: SPECIFICATIONS AND APPROVAL MATERIALS Noncombustible Materials for Merchant Vessels § 164.009-23 Factory... regular schedule. However, the Commander of the Coast Guard District in which a factory is located...

  10. 46 CFR 162.050-13 - Factory production and inspection.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 6 2010-10-01 2010-10-01 false Factory production and inspection. 162.050-13 Section....050-13 Factory production and inspection. (a) Equipment manufactured under Coast Guard approval must... any time to visit a factory where the equipment is manufactured to conduct an inspection of...

  11. 27 CFR 40.72 - Use of factory premises.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 2 2010-04-01 2010-04-01 false Use of factory premises... factory premises. (a) General. Unless otherwise authorized by the appropriate TTB officer as provided in § 40.47, the premises used by a manufacturer of tobacco products for his factory shall be...

  12. 27 CFR 45.33 - Return of shipment to factory.

    Science.gov (United States)

    2010-04-01

    ... factory. 45.33 Section 45.33 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE... factory. Tobacco products, and cigarette papers and tubes which have been removed, under this part, may be returned to the factory without internal revenue supervision. (72 Stat. 1418, as amended; 26 U.S.C. 5704)...

  13. Combined molecular and biochemical approach identifies Aspergillus japonicus and Aspergillus aculeatus as two species

    DEFF Research Database (Denmark)

    Parenicova, L.; Skouboe, P.; Frisvad, Jens Christian

    2001-01-01

    We examined nine Aspergillus japonicus isolates and 10 Aspergillus aculeatus isolates by using molecular and biochemical markers, including DNA sequences of the ITS1-5.8S rRNA gene-ITS2 region, restriction fragment length polymorphisms (RFLP), and secondary-metabolite profiles. The DNA sequence o...... probably represents a third taxon. This study provides unambiguous criteria for establishing the taxonomic positions of isolates of black aspergilli, which are important in relation to industrial use and legal protection of these organisms....

  14. Antibiotic Extraction as a Recent Biocontrol Method for Aspergillus Niger andAspergillus Flavus Fungi in Ancient Egyptian mural paintings

    Science.gov (United States)

    Hemdan, R. Elmitwalli; Fatma, Helmi M.; Rizk, Mohammed A.; Hagrassy, Abeer F.

    Biodeterioration of mural paintings by Aspergillus niger and Aspergillus flavus Fungi has been proved in different mural paintings in Egypt nowadays. Several researches have studied the effect of fungi on mural paintings, the mechanism of interaction and methods of control. But none of these researches gives us the solution without causing a side effect. In this paper, for the first time, a recent treatment by antibiotic "6 penthyl α pyrone phenol" was applied as a successful technique for elimination of Aspergillus niger and Aspergillus flavus. On the other hand, it is favorable for cleaning Surfaces of Murals executed by tembera technique from the fungi metabolism which caused a black pigments on surfaces.

  15. Simultaneous amyloglucosidase and exo-polygalacturonase production by Aspergillus niger using solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Jorge Alberto Vieira Costa

    2007-09-01

    Full Text Available Amyloglucosidase (AMG and exo-polygalacturonase (exo-PG were simultaneously produced by two different strains of Aspergillus niger in solid-state fermentation (SSF using defatted rice-bran as substrate. The effect of Aspergillus niger strain (t0005/007-2 and/or CCT 3312, inoculum type (spore suspension or fermented bran and addition of inducers (pectin and/or starch to the culture media was studied using a 3² x 2¹ factorial experimental design. The production of AMG and exo-PG was significantly affected by fungal strain and inoculum type but inducers had no effect. The maximum yields obtained were 1310 U/g dm for AMG using a spore suspension of A. niger CCT 3312 and 50.2 U/g dm for exo-PG production, using A. niger t0005/007-2 and fermented bran as inoculum. The yields obtained represented acceptable values in comparison with data available in the literature and indicated that defatted rice-bran was a good nutrient source.As enzimas amiloglicosidase (AMG e exo-poligalacturonase (exo-PG foram produzidas simultaneamente por duas cepas de Aspergillus niger, através de fermentação em estado sólido usando farelo de arroz desengordurado como substrato. Foram avaliados os efeitos da cepa de Aspergillus niger, tipo de inóculo e adição de indutores no meio de cultura, utilizando-se um planejamento experimental fracionário 3² x 2¹. O máximo rendimento obtido foi 1310 U/g ms para a produção de AMG e 50,2 U/g ms para a exo-PG. Comparando-se estes resultados com dados da literatura pode-se dizer que os rendimentos obtidos foram aceitáveis e indicam que o farelo de arroz desengordurado é uma boa fonte de nutrientes. A produção de AMG e exo-PG foi significativamente afetada pelas variáveis cepa de A. niger e tipo de inóculo, enquanto a variável indutor não apresentou influência significativa na produção destas enzimas.

  16. Assessment of noise in furniture factories

    Directory of Open Access Journals (Sweden)

    Alexandre Petusk Filipe

    2014-12-01

    Full Text Available Work safety is of great importance in all industrial activities. The Norm NR15 of the Brazilian law determines that the work environment be tailored to employees to minimize biological, ergonomic, physical, chemical risks and accidents. The aim of this study was to evaluate the exposure of intermittent noise in a daily dose to workers in furniture factories. Measurements were made with a sound level meter and dosimeter in a workday of eight hours in 14 furniture factories located in the South of Minas Gerais. Noise values ranged from 66.0 to 117.4 dB(A. At a dose of 8 working hours it was observed that the values exceed the tolerance limit of a unit, being from 2.76 to 30.52 for minimum and maximum units, respectively. Both noises and daily doses were superior than the values set by the NR-15-Annex 01 (BRASIL, 2008.

  17. The Physics of the B Factories

    Energy Technology Data Exchange (ETDEWEB)

    Bevan, A.J. [Queen Mary Univ., London (United Kingdom); Golob, B. [Ljubljana Univ. (Slovenia). Faculty of Mathematics and Physics; J. Stefan Institute, Ljubljana (Slovenia); Mannel, T. [Siegen Univ. (Germany). Theoretische Physik 1; Collaboration: BABAR Collaboration; Belle Collaboration; and others

    2014-11-15

    The inspiration for this book came from Fran@cois le Diberder. During his term as spokesperson for BABAR he laid down a vision for the two B Factory detector collaborations, BABAR and Belle, to work together on a book that would describe the methodologies used and physics results obtained by those experiments. A key ideal emphasized from the outset was that this book should be written from a pedagogical perspective; it should be of interest to the student and expert alike. This vision was presented during a BABAR collaboration meeting on the island of Elba in May 2008 and a follow up Belle collaboration meeting at KEK, with visiting colleagues from the BABAR collaboration, and was embraced by the community. A number of workshops involving people from the theoretical community as well as the two collaborations were held on four continents over the following years. The resulting book, ''The Physics of the B Factories'', is a testament to the way that this concept captured the zeitgeist on both sides of the Pacific Ocean. This book is divided into three parts, the first of which provides a brief description of the B Factories, including a short (though not exhaustive) historical perspective, as well as descriptions of the detectors, ancillary data acquisition systems and data (re)processing systems that were built by the two detector collaborations in the late 1990's. The second part of the book discusses tools and methods that are frequently used when analyzing the data collected. These range from details of low level reconstruction algorithms and abstract summaries of statistical methods to high level prescriptions used when evaluating systematic uncertainties on measurements of observables. The third part of the book is devoted to physics results. This includes sufficient theoretical discussion in order for the reader to understand the context of the work being described. We are indebted to our colleagues from the theoretical community who

  18. Persistence versus escape: Aspergillus terreus and Aspergillus fumigatus employ different strategies during interactions with macrophages.

    Directory of Open Access Journals (Sweden)

    Silvia Slesiona

    Full Text Available Invasive bronchopulmonary aspergillosis (IBPA is a life-threatening disease in immunocompromised patients. Although Aspergillus terreus is frequently found in the environment, A. fumigatus is by far the main cause of IBPA. However, once A. terreus establishes infection in the host, disease is as fatal as A. fumigatus infections. Thus, we hypothesized that the initial steps of disease establishment might be fundamentally different between these two species. Since alveolar macrophages represent one of the first phagocytes facing inhaled conidia, we compared the interaction of A. terreus and A. fumigatus conidia with alveolar macrophages. A. terreus conidia were phagocytosed more rapidly than A. fumigatus conidia, possibly due to higher exposure of β-1,3-glucan and galactomannan on the surface. In agreement, blocking of dectin-1 and mannose receptors significantly reduced phagocytosis of A. terreus, but had only a moderate effect on phagocytosis of A. fumigatus. Once phagocytosed, and in contrast to A. fumigatus, A. terreus did not inhibit acidification of phagolysosomes, but remained viable without signs of germination both in vitro and in immunocompetent mice. The inability of A. terreus to germinate and pierce macrophages resulted in significantly lower cytotoxicity compared to A. fumigatus. Blocking phagolysosome acidification by the v-ATPase inhibitor bafilomycin increased A. terreus germination rates and cytotoxicity. Recombinant expression of the A. nidulans wA naphthopyrone synthase, a homologue of A. fumigatus PksP, inhibited phagolysosome acidification and resulted in increased germination, macrophage damage and virulence in corticosteroid-treated mice. In summary, we show that A. terreus and A. fumigatus have evolved significantly different strategies to survive the attack of host immune cells. While A. fumigatus prevents phagocytosis and phagolysosome acidification and escapes from macrophages by germination, A. terreus is rapidly

  19. LAMPF: the meson factory. A LASL monograph

    Energy Technology Data Exchange (ETDEWEB)

    Allred, J.C.

    1977-08-01

    A general and simplified introduction to the entire concept of LAMPF is given in terms of its experimental capabilities. Parts of the current experimental program are used as illustrative examples. Topics discussed include: (1) the evolution of the meson factory; (2) accelerator construction; (3) strong focusing; (4) accelerator innovations at LAMPF; (5) photons and pions; (6) muons as nuclear probes; (7) nuclear chemistry; (8) radiobiology and medical applications; (9) radioisotope production; (10) materials testing; and (11) LAMPF management and users group. (PMA)

  20. Anaerobic digestion of cassava starch factory effluent.

    Science.gov (United States)

    Manilal, V B; Narayanan, C S; Balagopalan, C

    1990-06-01

    Biomethanation of cassava starch factory effluent in a batch digester produced 130 l biogas/kg dry matter with an average melthane content of 59%. About 63% COD was removed during 60 days. In semicontinuous digesters, gas production was 3251/kg dry matter with a retention time of 33,3 days giving a COD reduction of 50%. Size of starter inoculum was important for good biogasification of the effluent.

  1. 68Ga-triacetylfusarinine C and 68Ga-ferrioxamine E for Aspergillus infection imaging: uptake specificity in various microorganisms

    NARCIS (Netherlands)

    Petrik, M.; Haas, H. de; Laverman, P.; Schrettl, M.; Franssen, G.M.; Blatzer, M.; Decristoforo, C.

    2014-01-01

    (68)Ga-triacetylfusarinine C ((68)Ga-TAFC) and (68)Ga-ferrioxamine E ((68)Ga-FOXE) showed excellent targeting properties in Aspergillus fumigatus rat infection model. Here, we report on the comparison of specificity towards different microorganisms and human lung cancer cells (H1299).The in vitro up

  2. Crystallization and preliminary crystallographic analysis of endo-1,4-beta-xylanase I from Aspergillus niger

    NARCIS (Netherlands)

    Krengel, U; Rozeboom, HJ; Kalk, KH; Dijkstra, BW

    1996-01-01

    A family G xylanase from Aspergillus niger has been crystallized using the vapor-diffusion method. Several crystal forms could be obtained using various sodium salts as precipitants. Three of the crystal forms belong to space groups P2(1), P2(1)2(1)2(1) and P4(3) and have cell parameters of approxim

  3. Factorial complexity and Morally Debatable Behaviors

    Directory of Open Access Journals (Sweden)

    Grimaldo Muchotrigo, Mirian P.

    2011-12-01

    Full Text Available Currently, from the scientific and professional practice point of view, comes out the necessity to know more about moral permissiveness, as it seems to be an increase in “moral relativism”.. Because it, it this is important to have tools to collect valid and reliable information about moral in social situations defined as social and personal behavior issue. This paper presents a technical note of The Morally Debatable Behaviors Scale (MDBS from Harding & Phillips (1986, which was developed in USA and mainly focused on young people and adults. This technical note makes direct reference to a recent Latin American study (Merino & Grimaldo, 2010; this article focuses on the internal structure and the problems associated with evidences of factorial complexity among items of the MDBS. This means that the interpretation of scores is not factorially simple and could not be achieved by a conceptual distinction between the latent constructs applied to the study sample.. The results in the previous study of the factorial complexity leads the researcher to decide whether an instrument for measuring this aspect may contain a reasonable amount of complexity that is consistent with what is in reality, or look for the unidimensional and simple structure.

  4. An asymmetric B factory based on PEP

    Energy Technology Data Exchange (ETDEWEB)

    1991-02-01

    In this report we describe a design for a high-luminosity Asymmetric B Factory to be built in the PEP tunnel on the SLAC site. This proposal, a collaborative effort SLAC, LBL, and LLNL, is the culmination of more than two years of effort aimed at the design and construction of an asymmetric e{sup +}e{sup {minus}} collider capable of achieving a luminosity of L = 3 {times} 10{sup 33} cm{sup {minus}2} s{sup {minus}1}. The configuration adopted utilizes two storage rings, and electron ring operating at 9 GeV and a positron ring at 3.1 GeV, each with a circumference of 2200 m. The high-energy ring is an upgrade of the PEP storage ring at SLAC; all PEP magnets and most power supplies will be reused. The upgrade consists primarily of replacing the PEP vacuum chamber and RF system with newly designed versions optimized for the high-current environment of the B Factory. The low-energy ring will be newly constructed and will be situated atop the high-energy ring in the PEP tunnel. Utilities already installed in the PEP tunnel are largely sufficient to operate the two B Factory storage rings.

  5. Neutrinos: from the Workshop to the Factory

    CERN Multimedia

    2001-01-01

    Over the next 5 years much work will be done to reach a theoretical and practical description of a neutrino factory. How could this project turn out to be an interesting future option for CERN? Neutrino beams travelling from CERN to the Canary Islands? And to the Svalbard archipelago in Norway? Or even to the Pyhaesalmi Mine in Finland? Why neutrinos? And why so far? The answers provide one of CERN's next challenging options: the construction of a high-energy muon storage ring to provide neutrino beams. This project, nicknamed 'neutrino factory', now figures in CERN's middle term plan as a recognized and supported research and development project. International collaborations, with other European laboratories and also with America and Japan, are now being set up. Long baseline locations for neutrino oscillations studies at a CERN based neutrino factory. Early in its history, LEP established that there exist just three kinds of light neutrinos, those associated with the electron, muon, and tau leptons. For a...

  6. Mycotoxin production and predictive modelling kinetics on the growth of Aspergillus flavus and Aspergillus parasiticus isolates in whole black peppercorns (Piper nigrum L).

    Science.gov (United States)

    Yogendrarajah, Pratheeba; Vermeulen, An; Jacxsens, Liesbeth; Mavromichali, Evangelia; De Saeger, Sarah; De Meulenaer, Bruno; Devlieghere, Frank

    2016-07-02

    The growth and mycotoxin production of three Aspergillus flavus isolates and an Aspergillus parasiticus isolate were studied in whole black peppercorns (Piper nigrum L.) using a full factorial design with seven water activity (aw) (0.826-0.984) levels and three temperatures (22, 30 and 37°C). Growth rates and lag phases were estimated using linear regression. Diverse secondary models were assessed for their ability to describe the radial growth rate as a function of individual and combined effect of aw and temperature. Optimum radial growth rate ranged from 0.75±0.04 to 2.65±0.02mm/day for A. flavus and 1.77±0.10 to 2.50±0.10mm/day for A. parasiticus based on the Rosso cardinal estimations. Despite the growth failure of some isolates at marginal conditions, all the studied models showed good performance to predict the growth rates. Validation of the models was performed on independently derived data. The bias factors (0.73-1.03), accuracy factors (0.97-1.36) and root mean square error (0.050-0.278) show that the examined models are conservative predictors of the colony growth rate of both fungal species in black peppers. The Rosso cardinal model can be recommended to describe the individual aw effect while the extended Gibson model was the best model for describing the combined effect of aw and temperature on the growth rate of both fungal species in peppercorns. Temperature optimum ranged from 30 to 33°C, while aw optimum was 0.87-0.92 as estimated by multi-factorial cardinal model for both species. The estimated minimum temperature and aw for A. flavus and A. parasiticus for growth were 11-16°C and 0.73-0.76, respectively, hence, achieving these conditions should be considered during storage to prevent the growth of these mycotoxigenic fungal species in black peppercorns. Following the growth study, production of mycotoxins (aflatoxins B1, B2, G1, G2, sterigmatocystin and O-methyl sterigmatocystin (OMST)) was quantified using LC-MS/MS. Very small

  7. Fulminant antiphospholipid antibody syndrome complicated by Aspergillus tracheobronchitis.

    Science.gov (United States)

    Yegneswaran Prakash, Peralam; Pandit, Vinay; Rao, Sugandhi P

    2012-01-01

    Aspergillus fumigatus is a filamentous mold that causes infections in patients who are inmmunocompromised. We report a case of Aspergillus tracheobronchitis in fulminant systemic lupus erythematosus case. Diagnosis with more invasive diagnostic procedures & aggressive antifungal therapy is indicated at early stage.

  8. Evaluation of Aspergillus PCR Protocols for Testing Serum Specimens

    NARCIS (Netherlands)

    White, P.L.; Mengoli, C.; Bretagne, S.; Cuenca-Estrella, M.; Finnstrom, N.; Klingspor, L.; Melchers, W.J.G.; McCulloch, E.; Barnes, R.A.; Donnelly, J.P.; Loeffler, J.

    2011-01-01

    A panel of human serum samples spiked with various amounts of Aspergillus fumigatus genomic DNA was distributed to 23 centers within the European Aspergillus PCR Initiative to determine analytical performance of PCR. Information regarding specific methodological components and PCR performance was re

  9. Nationwide Surveillance of Azole Resistance in Aspergillus Diseases.

    Science.gov (United States)

    Vermeulen, Edith; Maertens, Johan; De Bel, Annelies; Nulens, Eric; Boelens, Jerina; Surmont, Ignace; Mertens, Anna; Boel, An; Lagrou, Katrien

    2015-08-01

    Aspergillus disease affects a broad patient population, from patients with asthma to immunocompromised patients. Azole resistance has been increasingly reported in both clinical and environmental Aspergillus strains. The prevalence and clinical impact of azole resistance in different patient populations are currently unclear. This 1-year prospective multicenter cohort study aimed to provide detailed epidemiological data on Aspergillus resistance among patients with Aspergillus disease in Belgium. Isolates were prospectively collected in 18 hospitals (April 2011 to April 2012) for susceptibility testing. Clinical and treatment data were collected with a questionnaire. The outcome was evaluated to 1 year after a patient's inclusion. A total of 220 Aspergillus isolates from 182 patients were included. The underlying conditions included invasive aspergillosis (n = 122 patients), allergic bronchopulmonary aspergillosis (APBA) (n = 39 patients), chronic pulmonary aspergillosis (n = 10 patients), Aspergillus bronchitis (n = 7 patients), and aspergilloma (n = 5 patients). The overall azole resistance prevalence was 5.5% (95% confidence interval [CI] 2.8 to 10.2%) and was 7.0% (4/57; 95% CI, 2.3 to 17.2%) in patients with APBA, bronchitis, aspergilloma, or chronic aspergillosis and 4.6% in patients with invasive aspergillosis (5/108; 95% CI, 1.7 to 10.7%). The 6-week survival in invasive aspergillosis was 52.5%, while susceptibility testing revealed azole resistance in only 2/58 of the deceased patients. The clinical impact of Aspergillus fumigatus resistance was limited in our patient population with Aspergillus diseases.

  10. Aspergillus fumigatus conidial melanin modulates host cytokine response

    NARCIS (Netherlands)

    L.Y.A. Chai (Louis); M.G. Netea (Mihai); J. Sugui (Janyce); A.G. Vonk (Alieke); W.W.J. van de Sande (Wendy); A. Warris (Adilia); K.J. Kwon-Chung (Kyung); B. Jan Kullberg (Bart)

    2010-01-01

    textabstractMelanin biopigments have been linked to fungal virulence. Aspergillus fumigatus conidia are melanised and are weakly immunogenic. We show that melanin pigments on the surface of resting Aspergillus fumigatus conidia may serve to mask pathogen-associated molecular patterns (PAMPs)-induced

  11. Aspergillus fumigatus conidial melanin modulates host cytokine response.

    NARCIS (Netherlands)

    Chai, L.; Netea, M.G.; Sugui, J.; Vonk, A.G.; Sande, W.W. van de; Warris, A.; Kwon-Chung, K.J.; Kullberg, B.J.

    2010-01-01

    Melanin biopigments have been linked to fungal virulence. Aspergillus fumigatus conidia are melanised and are weakly immunogenic. We show that melanin pigments on the surface of resting Aspergillus fumigatus conidia may serve to mask pathogen-associated molecular patterns (PAMPs)-induced cytokine re

  12. Fatal coinfection with Legionella pneumophila serogroup 8 and Aspergillus fumigatus.

    Science.gov (United States)

    Guillouzouic, Aurélie; Bemer, Pascale; Gay-Andrieu, Françoise; Bretonnière, Cédric; Lepelletier, Didier; Mahé, Pierre-Joachim; Villers, Daniel; Jarraud, Sophie; Reynaud, Alain; Corvec, Stéphane

    2008-02-01

    Legionella pneumophila is an important cause of community-acquired and nosocomial pneumonia. We report on a patient who simultaneously developed L. pneumophila serogroup 8 pneumonia and Aspergillus fumigatus lung abscesses. Despite appropriate treatments, Aspergillus disease progressed with metastasis. Coinfections caused by L. pneumophila and A. fumigatus remain exceptional. In apparently immunocompetent patients, corticosteroid therapy is a key risk factor for aspergillosis.

  13. Discovery of novel secondary metabolites in Aspergillus aculeatus

    DEFF Research Database (Denmark)

    Petersen, Lene Maj; Holm, Dorte Koefoed; Gotfredsen, Charlotte Held

    2012-01-01

    of the industrially important black Aspergillus Aspergillus aculeatus by UHPLC-DAD-HRMS has identified several SMs already known from this organism. However, several compounds could not be unambiguously dereplicated wherefore some have been selected, purified and structure elucidated by 1D and 2D NMR spectroscopy...

  14. Aspergillus flavus myositis in a patient after liver transplantation.

    NARCIS (Netherlands)

    Li, D.M.; Xiu, D.R.; Li, R.Y.; Samson, R.A.; de Hoog, G.S.; Wang, D.L.

    2008-01-01

    We describe the first case of Aspergillus myositis caused by Aspergillus flavus in a liver transplant patient. The patient was a 43-year-old man who underwent liver transplantation because of end-stage hepatic cirrhosis. He experienced pain in his left calf two months after the operation. Nodules wi

  15. Arabinase induction and carbon catabolite repression in Aspergillus niger and Aspergillus nidulans.

    NARCIS (Netherlands)

    Veen, van der P.

    1995-01-01

    The first aim of this thesis was to get a better understanding of the properties and the induction features of arabinan degrading enzymes and enzymes involved in the intracellular L-arabinose catabolic pathway in Aspergillus niger. The second aim was to understand the which role carbon catabolite re

  16. A comparison of Agrobacterium-mediated transformation and protoplast-mediated transformation with CRISPR-Cas9 and bipartite gene targeting substrates, as effective gene targeting tools for Aspergillus carbonarius.

    Science.gov (United States)

    Weyda, István; Yang, Lei; Vang, Jesper; Ahring, Birgitte K; Lübeck, Mette; Lübeck, Peter S

    2017-04-01

    In recent years, versatile genetic tools have been developed and applied to a number of filamentous fungi of industrial importance. However, the existing techniques have limitations when it comes to achieve the desired genetic modifications, especially for efficient gene targeting. In this study, we used Aspergillus carbonarius as a host strain due to its potential as a cell factory, and compared three gene targeting techniques by disrupting the ayg1 gene involved in the biosynthesis of conidial pigment in A. carbonarius. The absence of the ayg1 gene leads to phenotypic change in conidia color, which facilitated the analysis on the gene targeting frequency. The examined transformation techniques included Agrobacterium-mediated transformation (AMT) and protoplast-mediated transformation (PMT). Furthermore, the PMT for the disruption of the ayg1 gene was carried out with bipartite gene targeting fragments and the recently adapted CRISPR-Cas9 system. All three techniques were successful in generating Δayg1 mutants, but showed different efficiencies. The most efficient method for gene targeting was AMT, but further it was shown to be dependent on the choice of Agrobacterium strain. However, there are different advantages and disadvantages of all three gene targeting methods which are discussed, in order to facilitate future approaches for fungal strain improvements.

  17. DECOLORIZATION OF BLUE 13 WITH ASPERGILLUS NIVEUS AND FUSARIUM MONILIFORME

    Directory of Open Access Journals (Sweden)

    Hülya KARACA

    2012-02-01

    Full Text Available Depending on the changing world trends has been increasing interest in colored products. In the textile industry, to give the blue color to the textiles the heavily used Ambifiks Blue H5R (Blue 13 dye. This dye has been given from plants in the environment of the receiving water to results from the toxic effects. For reduction of this toxic effect has been isolated Aspergillus niveus and Fusarium moniliforme from porsuk Stream were invastigated because of decolorizasyon capabilities with different physiological conditions (pH, temprature, agitate condition. For A. niveus results have been obtained up to nearly %95 at pH 3, 35°C and 120-150 rpm. For F. moniliforme results have been obtained up to nearly %95 at pH 7, 20°C ve 120 rpm. For A. niveus decolorisation results were close to dead cells and live cells.Dead cells decolorisation were about %90 for F. moniliforme and live cells decolorisation were about %75.

  18. In-Factory Learning - Qualification For The Factory Of The Future

    Science.gov (United States)

    Quint, Fabian; Mura, Katharina; Gorecky, Dominic

    2015-07-01

    The Industry 4.0 vision anticipates that internet technologies will find their way into future factories replacing traditional components by dynamic and intelligent cyber-physical systems (CPS) that combine the physical objects with their digital representation. Reducing the gap between the real and digital world makes the factory environment more flexible, more adaptive, but also more complex for the human workers. Future workers require interdisciplinary competencies from engineering, information technology, and computer science in order to understand and manage the diverse interrelations between physical objects and their digital counterpart. This paper proposes a mixed-reality based learning environment, which combines physical objects and visualisation of digital content via Augmented Reality. It uses reality-based interaction in order to make the dynamic interrelations between real and digital factory visible and tangible. We argue that our learning system does not work as a stand-alone solution, but should fit into existing academic and advanced training curricula.

  19. Confirmatory factorial analysis of TEOSQp / Análise factorial confirmatória do TEOSQp

    Directory of Open Access Journals (Sweden)

    Helder M. Fernandes

    2010-01-01

    Full Text Available The present research assessed the reliability, factorial validity and measurement invariance (by gender of the Portuguese version of TEOSQ –Task and Ego Orientation in Sport Questionnaire (Fonseca & Biddle, 2001. Data were collected from 1010 physical education students with a mean age of 15.42 ( SD=1.91. Factorial and invariance measurements were tested via confirmatory factorial analysis. Results supported internal consistency for the two proposed subscales (task and ego. Based on modification indices and theoretical justification the TEOSQ p was reduced to 12 items with better goodness-of-fit indices for the oblique model. The results of gender invariance did not provide full empirical support to the multi-group equivalence assumption, being suggested that TEOSQ p does not measure in the same way goal orientations for boys (orthogonal model and girls (oblique model. In light of these results, conceptual, empirical and practical issues were discussed.

  20. Reduction of aflatoxin production by Aspergillus flavus and Aspergillus parasiticus in interaction with Streptomyces.

    Science.gov (United States)

    Verheecke, C; Liboz, T; Anson, P; Diaz, R; Mathieu, F

    2015-05-01

    The aim of this study is to investigate aflatoxin gene expression during Streptomyces-Aspergillus interaction. Aflatoxins are carcinogenic compounds produced mainly by Aspergillus flavus and Aspergillus parasiticus. A previous study has shown that Streptomyces-A. flavus interaction can reduce aflatoxin content in vitro. Here, we first validated this same effect in the interaction with A. parasiticus. Moreover, we showed that growth reduction and aflatoxin content were correlated in A. parasiticus but not in A. flavus. Secondly, we investigated the mechanisms of action by reverse-transcriptase quantitative PCR. As microbial interaction can lead to variations in expression of household genes, the most stable [act1, βtub (and cox5 for A. parasiticus)] were chosen using geNorm software. To shed light on the mechanisms involved, we studied during the interaction the expression of five genes (aflD, aflM, aflP, aflR and aflS). Overall, the results of aflatoxin gene expression showed that Streptomyces repressed gene expression to a greater level in A. parasiticus than in A. flavus. Expression of aflR and aflS was generally repressed in both Aspergillus species. Expression of aflM was repressed and was correlated with aflatoxin B1 content. The results suggest that aflM expression could be a potential aflatoxin indicator in Streptomyces species interactions. Therefore, we demonstrate that Streptomyces can reduce aflatoxin production by both Aspergillus species and that this effect can be correlated with the repression of aflM expression.

  1. Antifungal activity of Oleoresins used in meat industry on some toxigenic Aspergillus spp.

    Directory of Open Access Journals (Sweden)

    Šošo Vladislava M.

    2013-01-01

    Full Text Available Different spice oleoresins are widely used in meat industry. They contribute to the specific aroma and flavor of the end products, but they have also been reported to have strong antimicrobial activity. These properties open a plenty of possibilities to be used for defining the specific sensory profile of the product but also as natural food preservatives. This paper focuses on the antifungal activity of four oleoresins against different foodborne toxigenic Aspergillus species. Oleoresins used in the experiments were cayenne pepper, black pepper, garlic and rosemary oleoresins, and they were tested against following Aspergillus species: A. clavatus, A. flavus, A. fumigatus, A. niger, A. ochraceus and A. versicolor. Antifungal activity was tested using microtitre-plate-based assay incorporating resazurin as an indicator of cell growth and broth microdilution-method. [Projekat Ministarstva nauke Republike Srbije, br. III46009

  2. Secretome data from Trichoderma reesei and Aspergillus niger cultivated in submerged and sequential fermentation methods

    Directory of Open Access Journals (Sweden)

    Camila Florencio

    2016-09-01

    Full Text Available The cultivation procedure and the fungal strain applied for enzyme production may influence levels and profile of the proteins produced. The proteomic analysis data presented here provide critical information to compare proteins secreted by Trichoderma reesei and Aspergillus niger when cultivated through submerged and sequential fermentation processes, using steam-explosion sugarcane bagasse as inducer for enzyme production. The proteins were organized according to the families described in CAZy database as cellulases, hemicellulases, proteases/peptidases, cell-wall-protein, lipases, others (catalase, esterase, etc., glycoside hydrolases families, predicted and hypothetical proteins. Further detailed analysis of this data is provided in “Secretome analysis of Trichoderma reesei and Aspergillus niger cultivated by submerged and sequential fermentation process: enzyme production for sugarcane bagasse hydrolysis” C. Florencio, F.M. Cunha, A.C Badino, C.S. Farinas, E. Ximenes, M.R. Ladisch (2016 [1].

  3. Prizes awarded in fiscal 1999 by the Minister for International Trade and Industry on factories having applied excellent energy management. Energy conservation by installing fuel cell power generation facilities utilizing methane gas generated from waste water treatment plants / Improvements toward a clean room and energy saving air conditioning system; 1999 nendo energy kanri yuryo kojo tsusho sangyo daijin hyosho jusho. 1999 nendo shigen energy sho chokan hyosho jusho

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2000-04-01

    In order to achieve energy conservation in a waste water treatment plant in a brewery factories, an anaerobic treatment facility was introduced, and a fuel cell power generation facility effectively utilizing methane gas generated from the anaerobic waste water treatment plant was installed. This has resulted in large reduction in the operating number of blowers in the waste water treatment facility of activated sludge system. In addition, electric power, steam, and hot water generated from the fuel cells are effectively utilized as the factory utility. In energy conservation in an optical communication device manufacturing factory, the fan filter unit system was adopted, having been changed from the circulation air conditioner, a conventional type air conditioning system. The present system is a circulation system integrating the fan with the filter, making it possible to circulate air in the room to perform control of temperature, humidity, and dust in a clean room. Thus, the system has eliminated the circulating air conditioner, and reduced the air conditioner capacity by 42% and power consumption by 58.6% compared with those in the conventional circulation system. (NEDO)

  4. ADOPTING SELECTED HYDROGEN BONDING AND IONIC INTERACTIONS FROM ASPERGILLUS FUMIGATUS PHYTASE STRUCTURE IMPROVES THE THERMOSTABILITY OF ASPERGILLUS NIGER PHYA PHYTASE

    Science.gov (United States)

    Although it has been widely used as a feed supplement to reduce manure phosphorus pollution of swine and poultry, Aspergillus niger PhyA phytase is unable to withstand heat inactivation during feed pelleting. Crystal structure comparisons with its close homolog, the thermostable Aspergillus fumigatu...

  5. Aspergillus triggers phenazine production in Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Jensen, Britt Guillaume; Jelsbak, Lars; Søndergaard, Ib

    Aspergillus species. Methods: A suspension of fungal spores was streaked onto WATM agar plates. After 24 hours incubation at 37 °C, a P. aeruginosa overnight culture was streaked out perpendicular to the fungal streak. The plates were incubated at 37 °C for five days, examined and plugs were extracted...... for HPLC-DAD and HPLC-DAD-MS analysis. Results: P. aeruginosa PAO1 suppressed growth of A. fumigatus, A. niger, A. flavus, A. oryzae, A. terreus and Emericella nidulans. HPLC and HPLC-DAD-MS results showed an increase in phenazine-1-carboxylic acid and phenazine-1-carboxamide production by P. aeruginosa...

  6. Suppression of Aspergillus by Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Jensen, Britt Guillaume; Jelsbak, Lars; Søndergaard, Ib

    culture plates. After 24 hours incubation at 37 °C, a P. aeruginosa overnight culture diluted to 108 CFU/ml was streaked out perpendicular to the fungal streak. The plates were incubated at 37 °C for 5 days, examined and plugs were extracted for HPLC and LC-DAD-MS analysis. Results: P. aeruginosa PAO1...... suppressed growth of A. fumigatus, A. niger, A. flavus, A. oryzae, A. terreus and E. nidulans. HPLC and LC-DAD-MS results showed an increase in phenazine-1-carboxylic acid and phenazine-1-carboxamide production by P. aeruginosa in the contact area of Aspergillus. Different quinolones were also identified...

  7. Object Classification at the Nearby Supernova Factory

    Energy Technology Data Exchange (ETDEWEB)

    Aragon, Cecilia R.; Bailey, Stephen; Aragon, Cecilia R.; Romano, Raquel; Thomas, Rollin C.; Weaver, B. A.; Wong, D.

    2007-12-21

    We present the results of applying new object classification techniques to the supernova search of the Nearby Supernova Factory. In comparison to simple threshold cuts, more sophisticated methods such as boosted decision trees, random forests, and support vector machines provide dramatically better object discrimination: we reduced the number of nonsupernova candidates by a factor of 10 while increasing our supernova identification efficiency. Methods such as these will be crucial for maintaining a reasonable false positive rate in the automated transient alert pipelines of upcoming large optical surveys.

  8. High-gravity brewing utilizing factorial design

    OpenAIRE

    Almeida,R. B.; J. B. Almeida e Silva; Lima,U. A.; Assis,A. N.

    2000-01-01

    A number of factors can influence the behavior of yeast during fermentation. Some of these factors (initial wort concentration, initial pH and percentage of corn syrup in the composition of the wort) were studied in order to determine their influence on the productivity of fermentation. Fermentations were carried out at 25ºC utilizing a 2³ factorial design of these factors. The results showed that the percentage of corn syrup had no influence on process productivity, whereas initial pH and es...

  9. Tau/Charm Factory Accelerator Report

    CERN Document Server

    Biagini, M E; Boscolo, M; Chiarucci, A; Cimino, R; Clozza, A; Drago, A; Guiducci, S; Ligi, C; Mazzitelli, G; Ricci, R; Sanelli, C; Serio, M; Stella, A; Tomassini, S; Bini, S; Cioeta, F; Cittadino, D; D'Agostino, M; Del Franco, M; Piane, A Delle; Di Pasquale, E; Frascadore, G; Gazzana, S; Gargana, R; Incremona, S; Michelotti, A; Sabbatini, L; Schillaci, G; Sedita, M; Raimondi, P; Petronzio, R; Paoloni, E; Liuzzo, S M; Carmignani, N; Pivi, M

    2013-01-01

    The present Report concerns the current status of the Italian Tau/Charm accelerator project and in particular discusses the issues related to the lattice design, to the accelerators systems and to the associated conventional facilities. The project aims at realizing a variable energy Flavor Factory between 1 and 4.6 GeV in the center of mass, and succeeds to the SuperB project from which it inherits most of the solutions proposed in this document. The work comes from a cooperation involving the INFN Frascati National Laboratories accelerator experts, the young newcomers, mostly engineers, of the Cabibbo Lab consortium and key collaborators from external laboratories.

  10. Flavichalasines A–M, cytochalasan alkaloids from Aspergillus flavipes

    Science.gov (United States)

    Wei, Guangzheng; Tan, Dongdong; Chen, Chunmei; Tong, Qingyi; Li, Xiao-Nian; Huang, Jinfeng; Liu, Junjun; Xue, Yongbo; Wang, Jianping; Luo, Zengwei; Zhu, Hucheng; Zhang, Yonghui

    2017-01-01

    Two new tetracyclic cytochalasans, flavichalasines A and B (1 and 2), three new pentacyclic cytochalasans, flavichalasines C–E (3–5), and eight new tricyclic cytochalasans, flavichalasines F–M (6–13), together with eight known analogues (14–21), were isolated from the solid culture of Aspergillus flavipes. Structures of these new compounds were elucidated on the basis of extensive spectroscopic analyses including 1D, 2D NMR and HRESIMS data. Their absolute configurations were determined by comparison of their experimental ECD with either computed ECD or experimental ECD spectrum of known compound. The structure and absolute configuration of 2 were further determined by X-ray crystallographic diffraction. Flavichalasine A (1) represents the first example of cytochalasan with a terminal double bond at the macrocyclic ring and flavichalasine E (5) is the only cytochalasan with an α-oriented oxygen-bridge in D ring. These new compounds were evaluated for their cytotoxic activities against seven human cancer cell lines, of which, 6 and 14 displayed moderate inhibitory activities against tested cell lines. In addition, compounds 6 and 14 induced apoptosis of HL60 cells by activation of caspase-3 and degradation of PARP. PMID:28205583

  11. Scleral buckle infection with aspergillus flavus

    Directory of Open Access Journals (Sweden)

    Bouhaimed Manal

    2008-01-01

    Full Text Available Purpose: To present a case of scleral buckle infection with Aspergillus flavus in a tertiary eye center in Saudi Arabia. Methods: A retrospective case report of a 28-year-old Saudi male who presented with a six-month history of conjunctival injection and discharge from the left eye which had undergone uncomplicated conventional retinal detachment surgery, at the King Khaled Eye Specialist Hospital in Riyadh, Saudi Arabia, in the form of cryopexy, subretinal fluid drainage and scleral buckle (grooved segmental sponge and circumferential band with sleeve for a macula on retinal detachment four years earlier. A diagnosis of infected extruded scleral buckle was made and the buckle was removed. Results: The infected scleral buckle was removed under local anesthesia with administration of sub-conjunctival irrigation of 50 mg solution of Vancomycin, and sub-conjunctival injection of 25mg of Vancomycin. Post operative microbiological studies revealed infection with silver staining of moderate Aspergillus flavus hyphae. Visual acuity of the left eye improved from 20/200 before surgery to 20/60 in the two years follow-up visit. Conclusion: This case report indicates the importance of considering infection with multiple organisms - including fungal ones - in cases of scleral buckle infections in our population.

  12. Aspergillus terreus recovered from a corneal scraping.

    Science.gov (United States)

    Campbell, Suzanne

    2014-01-01

    A 52 year old, healthy male presented to his optometrist complaining of redness and irritation in the right eye. A foreign body was removed from the eye. The patient was started on ophthalmic solutions of vigamox and systane. At 48 hours, the patient reported increased redness, limited vision, and yellow discharge from the eye. The patient was referred to an ophthalmologist for further evaluation. Physical assessment revealed a superlative central infiltrate (extreme, centrally located injury that had permeated the cornea), diffuse corneal haze, and edema with a 3- to 4+ conjunctival injection and a 1 millimeter hypopyon (an effusion of pus into the anterior chamber of the eye). Corneal scrapings were collected for aerobic and anaerobic bacterial and fungal cultures. The patient was then prescribed. vancomycin, tobramycin, and natamycin ophthalmic eyedrops. On day three, fungal culture results indicated possible fungal forms seen. On day 12, results from the fungal culture of the corneal scraping revealed the causative agent to be Aspergillus terreus. Voriconazole eyedrops were added to the treatment regimen and continued for 10 weeks. The physician order for a fungal culture as well as laboratory data providing the final identification of Aspergillus terreus and laboratory comments indicating an elevated minimum inhibitory concentration (MIC) (> 2 microg/mL) to amphotericin B is associated with treatment failure positively impacted the patient outcome. After completion of the treatment regimen, a photo-therapeutic keratectomy (PTK) was performed in an attempt to remove the dense corneal scarring caused by the fungal infection.

  13. Endopolygalacturonase and pectinesterase from Aspergillus Niger

    Directory of Open Access Journals (Sweden)

    Adriana Lozano Química

    2011-12-01

    Full Text Available Endo-polymethylgalacturonase (endo-PMG EC 3.2.1.41 was produced by submerged fermentation using a local Aspergillus niger strain's Aspergillus niger spores (1xl05 spores/ml medium as inoculum. The only carbon source present in the fermentation medium was commercial citrus pectin. The enzyme was partially purified by precipitation with 20% ammonium sulphate, followed by chromatography in Sephadex G-75 and DEAE Sephadex A-50 columns. Pectinesterase yield was low in those fermentation conditions used. Fermentation crude extract presented two pH values (endo-PMG optimum pH 4.5 and 6.3. The enzymes were stable at pH values between 2.0 and 10.0. Optimum temperature was in the 40°C-45°C range. Enzymes became totally inactive at 70°C. It was also found that sodium ion concentrations greater than 0.1M inhibited endo-PMG. 

  14. An inventory of the Aspergillus niger secretome by combining in silico predictions with shotgun proteomics data

    NARCIS (Netherlands)

    Braaksma, M.; Martens-Uzunova, E.S.; Punt, P.J.; Schaap, P.J.

    2010-01-01

    Background: The ecological niche occupied by a fungal species, its pathogenicity and its usefulness as a microbial cell factory to a large degree depends on its secretome. Protein secretion usually requires the presence of a N-terminal signal peptide (SP) and by scanning for this feature using avail

  15. An inventory of the Aspergillus niger secretome by combining in silico predictions with shotgun proteomics data

    NARCIS (Netherlands)

    Braaksma, M.; Martens-Uzunova, E.S.; Punt, P.J.; Schaap, P.J.

    2010-01-01

    BACKGROUND: The ecological niche occupied by a fungal species, its pathogenicity and its usefulness as a microbial cell factory to a large degree depends on its secretome. Protein secretion usually requires the presence of a N-terminal signal peptide (SP) and by scanning for this feature using avail

  16. The Skateboard Factory: Curriculum by Design--Oasis Skateboard Factory Q&A with Craig Morrison

    Science.gov (United States)

    Pearson, George

    2012-01-01

    Since its opening three years ago, Oasis Skateboard Factory (OSF), founded by teacher Craig Morrison, has attracted considerable media exposure and received a Ken Spencer Award from the CEA for its innovative program. OSF is one of three programs offered by Oasis Alternative Secondary School, one of 22 alternative secondary schools of the Toronto…

  17. Chronological aging in conidia of pathogenic Aspergillus: Comparison between species.

    Science.gov (United States)

    Oliveira, Manuela; Pereira, Clara; Bessa, Cláudia; Araujo, Ricardo; Saraiva, Lucília

    2015-11-01

    Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus and Aspergillus niger are common airborne fungi, and the most frequent causative agents of human fungal infections. However, the resistance and lifetime persistence of these fungi in the atmosphere, and the mechanism of aging of Aspergillus conidia are unknown.With this work, we intended to study the processes underlying conidial aging of these four relevant and pathogenic Aspergillus species. Chronological aging was therefore evaluated in A. fumigatus, A. flavus, A. terreus and A. niger conidia exposed to environmental and human body temperatures. The results showed that the aging process in Aspergillus conidia involves apoptosis,with metacaspase activation, DNA fragmentation, and reactive oxygen species production, associated with secondary necrosis. Distinct results were observed for the selected pathogenic species. At environmental conditions, A. niger was the species with the highest resistance to aging, indicating a higher adaption to environmental conditions, whereas A. flavus followed by A. terreus were the most sensitive species. At higher temperatures (37 °C), A. fumigatus presented the longest lifespan, in accordance with its good adaptation to the human body temperature. Altogether,with this work new insights regarding conidia aging are provided, which may be useful when designing treatments for aspergillosis.

  18. Optimization of a furniture factory layout

    Directory of Open Access Journals (Sweden)

    Tadej Kanduč

    2015-03-01

    Full Text Available This paper deals with the problem of optimizing a factory floor layout in a Slovenian furniture factory. First, the current state of the manufacturing system is analyzed by constructing a discrete event simulation (DES model that reflects the manufacturing processes. The company produces over 10,000 different products, and their manufacturing processes include approximately 30,000 subprocesses. Therefore, manually constructing a model to include every subprocess is not feasible. To overcome this problem, a method for automated model construction was developed to construct a DES model based on a selection of manufacturing orders and relevant subprocesses. The obtained simulation model provided insight into the manufacturing processes and enable easy modification of model parameters for optimizing the manufacturing processes. Finally, the optimization problem was solved: the total distance the products had to traverse between machines was minimized by devising an optimal machine layout. With the introduction of certain simplifications, the problem was best described as a quadratic assignment problem. A novel heuristic method based on force-directed graph drawing algorithms was developed. Optimizing the floor layout resulted in a significant reduction of total travel distance for the products.

  19. Muon Acceleration Concepts for Future Neutrino Factory

    Energy Technology Data Exchange (ETDEWEB)

    Bogacz, Slawomir Alex [Thomas Jefferson National Accelerator Facility (TJNAF), Newport News, VA (United States)

    2016-05-01

    Here, we summarize current state of concept for muon acceleration aimed at future Neutrino Factory. The main thrust of these studies was to reduce the overall cost while maintaining performance through exploring interplay between complexity of the cooling systems and the acceptance of the accelerator complex. To ensure adequate survival of the short-lived muons, acceleration must occur at high average gradient. The need for large transverse and longitudinal acceptances drives the design of the acceleration system to initially low RF frequency, e.g. 325 MHz, and then increased to 650 MHz, as the transverse size shrinks with increasing energy. High-gradient normal conducting RF cavities at these frequencies require extremely high peak-power RF sources. Hence superconducting RF (SRF) cavities are chosen. Here, we considered two cost effective schemes for accelerating muon beams for a stagable Neutrino Factory: Exploration of the so-called 'dual-use' linac concept, where the same linac structure is used for acceleration of both H- and muons and alternatively, the SRF efficient design based on multi-pass (4.5) 'dogbone' RLA, extendable to multi-pass FFAG-like arcs.

  20. Carbonate factories: A conundrum in sedimentary geology

    Science.gov (United States)

    Pomar, L.; Hallock, P.

    2008-03-01

    by the end-Permian extinctions, excess photosynthesis by phytoplankton and microbial assemblages in surface waters, induced by moderately high CO 2 and temperature during the Early Mesozoic, supported proliferation of non-tissular metazoans (e.g., sponges) and heterotrophic bacteria at the sea floor. Metabolic activity by those microbes, especially sulfate reduction, resulted in abundant biologically-induced geochemical carbonate precipitation on and within the sea floor. For example, with the opening of Tethyan seaways during the Triassic, massive sponge/microbe boundstones (the benthic automicrite factory) formed steep, massive and thick progradational slopes and, locally, mud-mounds. As tectonic processes created shallow epicontinental seas, photosynthesis drove lime-mud precipitation in the illuminated zone of the water column. The resulting neritic lime-mud component of the shallow-water carbonate factory became predominant during the Jurassic, paralleling the increase in atmospheric pCO 2, while the decreasing importance of the benthic automicrite factory parallels the diversification of calcifying metazoans, phytoplankton and zooplankton. With atmospheric pCO 2 declining through the Cretaceous, the potential habitats for neritic lime-mud precipitation declined. At the same time, peak oceanic Ca 2+ concentrations promoted biotically-controlled calcification by the skeletal factory. With changes produced by extinctions and turnovers at the Cretaceous-Tertiary boundary, adaptations to decreasing Ca 2+ and pCO 2, coupled with increasing global temperature gradients (i.e., high-latitude and deep-water cooling), and strategies that efficiently linked photosynthesis and calcification, promoted successive changes of the dominant skeletal factory through the Cenozoic: larger benthic foraminifers (protist-protist symbiosis) during the Paleogene, red algae during the Miocene and modern coral reefs (metazoan-protist symbiosis) since Late Miocene.

  1. Integrating PCLIPS into ULowell's Lincoln Logs: Factory of the future

    Science.gov (United States)

    Mcgee, Brenda J.; Miller, Mark D.; Krolak, Patrick; Barr, Stanley J.

    1990-01-01

    We are attempting to show how independent but cooperating expert systems, executing within a parallel production system (PCLIPS), can operate and control a completely automated, fault tolerant prototype of a factory of the future (The Lincoln Logs Factory of the Future). The factory consists of a CAD system for designing the Lincoln Log Houses, two workcells, and a materials handling system. A workcell consists of two robots, part feeders, and a frame mounted vision system.

  2. Transformation of Litchi Pericarp-Derived Condensed Tannin with Aspergillus awamori

    Directory of Open Access Journals (Sweden)

    Sen Lin

    2016-07-01

    Full Text Available Condensed tannin is a ubiquitous polyphenol in plants that possesses substantial antioxidant capacity. In this study, we have investigated the polyphenol extraction recovery and 2,2-diphenyl-1-picrylhydrazyl (DPPH scavenging activity of the extracted polyphenol after litchi pericarp is treated with Aspergillus awamori, Aspergillus sojae or Aspergillus oryzae. We have further explored the activity of A. awamori in the formation of condensed tannin. The treatment of A. awamori appeared to produce the highest antioxidant activity of polyphenol from litchi pericarp. Further studies suggested that the treatment of A. awamori releases the non-extractable condensed tannin from cell walls of litchi pericarp. The total extractable tannin in the litchi pericarp residue after a six-time extraction with 60% ethanol increased from 199.92 ± 14.47–318.38 ± 7.59 μg/g dry weight (DW after the treatment of A. awamori. The ESI-TOF-MS and HPLC-MS2 analyses further revealed that treatment of A. awamori degraded B-type condensed tannin (condensed flavan-3-ol via C4–C8 linkage, but exhibited a limited capacity to degrade the condensed tannin containing A-type linkage subunits (C4–C8 coupled C2–O–C7 linkage. These results suggest that the treatment of A. awamori can significantly improve the production of condensed tannin from litchi pericarp.

  3. [New natural products from the marine-derived Aspergillus fungi-A review].

    Science.gov (United States)

    Zhao, Chengying; Liu, Haishan; Zhu, Weiming

    2016-03-04

    Marine-derived fungi were the main source of marine microbial natural products (NPs) due to their complex genetic background, chemodiversity and high yield of NPs. According to our previous survey for marine microbial NPs from 2010 to 2013, Aspergillus fungi have received the most of attention among all the marine-derived fungi, which accounted for 31% NPs of the marine fungal origins. This paper reviewed the sources, chemical structures and bioactivites of all the 512 new marine NPs of Aspergillus fungal origins from 1992 to 2014. These marine NPs have diverse chemical structures including polyketides, fatty acids, sterols and terpenoids, alkaloids, peptides, and so on, 36% of which displayed bioactivities such as cytotoxicity, antimicrobial activity, antioxidant and insecticidal activity. Nitrogen compounds are the major secondary metabolites accounting for 52% NPs from the marine-derived Aspergillus fungi. Nitrogen compounds are also the class with the highest ratio of bioactive compounds, 40% of which are bioactive. Plinabulin, a dehydrodiketopiperazine derivative of halimide had been ended its phase II trial and has received its phase III study from the third quarter of 2015 for the treatment of advanced, metastatic non-small cell lung cancer.

  4. Aspergillus in the lung: diverse and coincident forms

    Energy Technology Data Exchange (ETDEWEB)

    Buckingham, Susan J.; Hansell, David M. [Department of Radiology, Royal Brompton Hospital, Sydney Street, London SW3 6NP (United Kingdom)

    2003-08-01

    Pulmonary disease caused by the fungus Aspergillus has traditionally been regarded as belonging to one of the following, apparently distinct, entities: saprophytic aspergilloma; allergic bronchopulmonary aspergillosis (ABPA); and invasive aspergillosis (IPA); which may be further categorised as angioinvasive, acute or chronic airway invasive [1]. It is not always obvious that there is overlap between these entities, and that in any given patient more than one Aspergillus-related pathological process can co-exist [2]. The aim of this article is to review the clinical and imaging features of the main categories of Aspergillus-related pulmonary disease and, in particular, to highlight the overlap between them. (orig.)

  5. Antifungal susceptibility profile of cryptic species of Aspergillus.

    Science.gov (United States)

    Alastruey-Izquierdo, Ana; Alcazar-Fuoli, Laura; Cuenca-Estrella, Manuel

    2014-12-01

    The use of molecular tools has led to the description of new cryptic species among different Aspergillus species complexes. Their frequency in the clinical setting has been reported to be between 10 and 15%. The susceptibility to azoles and amphotericin B of many of these species is low, and some of them, such as Aspergillus calidoustus or Aspergillus lentulus, are considered multi-resistant. The changing epidemiology, the frequency of cryptic species, and the different susceptibility profiles make antifungal susceptibility testing an important tool to identify the optimal antifungal agent to treat the infections caused by these species.

  6. PENGARUH RHIZOPUS ORYZAE DAN ASPERGILLUS ORYZAE TERHADAP KUALITAS KECAP

    Directory of Open Access Journals (Sweden)

    Dewi Sabita Slamet

    2012-11-01

    Full Text Available Telah diteliti pengganti fermentasi mikroorganisme Aspergillus oryzae Rhyzopus oryzae dan campuran Aspergillus dan Rhyzopus oryzae, dengan perendaman dalam larutan garam 20% dalam waktu yang berbeda terhadap kualitas kecap.Lamanya perendaman dalam larutan garam 20% yang berbeda menghasilkan kadar protein kecap yang berbeda. Aspergillus oryzae lebih baik dalam menghasilkan enzima protease dari pada Rhyzopus oryzae.Uji organoleptik menunjukkan perbedaan tidak bermakna dalam hal rasa maupun aroma antar kecap yang dibuat dengan strain jamur yang berlainan serta waktu perendaman yang berbeda. Untuk membuat kecap, sebaiknya dilakukan perendaman dalam larutan garam 20% selama 14 hari.

  7. Fumonisins in Aspergillus niger: Industrial and food aspects

    DEFF Research Database (Denmark)

    Frisvad, Jens Christian; Nielsen, Kristian Fog; Mogensen, Jesper

    Introduction: Fumonisins are toxic seconday metabolites from Fusarium verticillioides and other Fusaria, from Tolypocladium and Aspergillus niger 1,2. Being a generalist Aspergillus niger is the workhorse in a very large number of industrial applications, and is also a common contaminant in foods...... produced fumonisins in pure culture, so we tested whether they could produce fumonisins on citric acid production media in shake flasks, and they could indeed produce small amounts of fumonisins. Conclusions: Most strains of Aspergillus niger can produce fumonisins. In order to have entirely safe...

  8. Effect of Vitronectin Bound to Insulin-Like Growth Factor-I and Insulin-Like Growth Factor Binding Protein-3 on Porcine Enamel Organ-Derived Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Yoshinori Shinohara

    2012-01-01

    Full Text Available The aim of this paper was to determine whether the interaction between IGF, IGFBP, and VN modulates the functions of porcine EOE cells. Enamel organs from 6-month-old porcine third molars were dissociated into single epithelial cells and subcultured on culture dishes pretreated with VN, IGF-I, and IGFBP-3 (IGF-IGFBP-VN complex. The subcultured EOE cells retained their capacity for ameloblast-related gene expression, as shown by semiquantitative reverse transcription-polymerase chain reaction. Amelogenin expression was detected in the subcultured EOE cells by immunostaining. The subcultured EOE cells were then seeded onto collagen sponge scaffolds in combination with fresh dental mesenchymal cells and transplanted into athymic rats. After 4 weeks, enamel-dentin-like complex structures were present in the implanted constructs. These results show that EOE cells cultured on IGF-IGFBP-VN complex differentiated into ameloblasts-like cells that were able to secrete amelogenin proteins and form enamel-like tissues in vivo. Functional assays demonstrated that the IGF/IGFBP/VN complex significantly enhanced porcine EOE cell proliferation and tissue forming capacity for enamel. This is the first study to demonstrate a functional role of the IGF-IGFBP-VN complex in EOE cells. This application of the subculturing technique provides a foundation for further tooth-tissue engineering and for improving our understanding of ameloblast biology.

  9. Effect of vitronectin bound to insulin-like growth factor-I and insulin-like growth factor binding protein-3 on porcine enamel organ-derived epithelial cells.

    Science.gov (United States)

    Shinohara, Yoshinori; Tsuchiya, Shuhei; Hatae, Kazuo; Honda, Masaki J

    2012-01-01

    The aim of this paper was to determine whether the interaction between IGF, IGFBP, and VN modulates the functions of porcine EOE cells. Enamel organs from 6-month-old porcine third molars were dissociated into single epithelial cells and subcultured on culture dishes pretreated with VN, IGF-I, and IGFBP-3 (IGF-IGFBP-VN complex). The subcultured EOE cells retained their capacity for ameloblast-related gene expression, as shown by semiquantitative reverse transcription-polymerase chain reaction. Amelogenin expression was detected in the subcultured EOE cells by immunostaining. The subcultured EOE cells were then seeded onto collagen sponge scaffolds in combination with fresh dental mesenchymal cells and transplanted into athymic rats. After 4 weeks, enamel-dentin-like complex structures were present in the implanted constructs. These results show that EOE cells cultured on IGF-IGFBP-VN complex differentiated into ameloblasts-like cells that were able to secrete amelogenin proteins and form enamel-like tissues in vivo. Functional assays demonstrated that the IGF/IGFBP/VN complex significantly enhanced porcine EOE cell proliferation and tissue forming capacity for enamel. This is the first study to demonstrate a functional role of the IGF-IGFBP-VN complex in EOE cells. This application of the subculturing technique provides a foundation for further tooth-tissue engineering and for improving our understanding of ameloblast biology.

  10. Heterologous expression of the Aspergillus nidulans alcR-alcA system in Aspergillus niger.

    Science.gov (United States)

    Nikolaev, I; Mathieu, M; van de Vondervoort, P; Visser, J; Felenbok, B

    2002-10-01

    The inducible and strongly expressed alcA gene encoding alcohol dehydrogenase I from Aspergillus nidulans was transferred together with the activator gene alcR, in the industrial fungus Aspergillus niger. This latter organism does not possess an inducible alc system but has an endogenously constitutive lowly expressed alcohol dehydrogenase activity. The overall induced expression of the alcA gene was of the same order in both fungi, as monitored by alcA transcription, alcohol dehydrogenase activity and heterologous expression of the reporter enzyme, beta-glucuronidase. However, important differences in the pattern of alcA regulation were observed between the two fungi. A high basal level of alcA transcription was observed in A. niger resulting in a lower ratio of alcA inducibility. This may be due to higher levels of the physiological inducer of the alc regulon, acetaldehyde, from general metabolism in A. niger which differs from that of A. nidulans.

  11. Identification and toxigenic potential of the industrially important fungi, Aspergillus oryzae and Aspergillus sojae

    DEFF Research Database (Denmark)

    Jørgensen, Thomas R

    2007-01-01

    Mold strains belonging to the species Aspergillus oryzae and Aspergillus sojae are highly valued as koji molds in the traditional preparation of fermented foods, such as miso, sake, and shoyu, and as protein production hosts in modern industrial processes. A. oryzae and A. sojae are relatives....... sojae strains. Separation of A. oryzae and A. sojae from A. flavus and A. parasiticus, respectively, is inconsistent, and both morphologic and molecular evidence support conspecificity. The high degree of identity is reflected by the divergent identification of reference cultures maintained in culture...... collections. As close relatives of aflatoxin-producing wild molds, koji molds possess an aflatoxin gene homolog cluster. Some strains identified as A. oryzae and A. sojae have been implicated in aflatoxin production. Identification of a strain as A. oryzae or A. sojae is no guarantee of its inability...

  12. Review of the Neutrino Factory Muon Front End

    CERN Document Server

    Rogers, C

    2011-01-01

    Three major facilities have been proposed for the precision study of neutrino oscillation parameters, the Neutrino Factory, the Betabeam and the Superbeam. Of these the Neutrino Factory offers high precision measurement of oscillations parameters. The Neutrino Factory generates neutrinos by firing protons onto a target in order to produce pions. The pions decay to muons which are captured before being accelerated to 25 GeV and stored in racetrack-shaped rings where they decay to neutrinos. In this note the pion decay channel, longitudinal drift, adiabatic buncher, phase rotation and ionisation cooling system that make up the Neutrino Factory muon front end are reviewed.

  13. Methylthio-Aspochalasins from a Marine-Derived Fungus Aspergillus sp.

    Directory of Open Access Journals (Sweden)

    Ying Liu

    2014-09-01

    Full Text Available Two novel aspochalasins, 20-β-methylthio-aspochalsin Q (named as aspochalasin V, (1 and aspochalasin W (2, were isolated from culture broth of Aspergillus sp., which was found in the gut of a marine isopod Ligia oceanica. The structures were determined on the basis of NMR and mass spectral data analysis. This is the first report about methylthio-substituted aspochalasin derivatives. Cytotoxicity against the prostate cancer PC3 cell line and HCT116 cell line was assayed using the MTT method. Apochalasin V showed moderate activity at IC50 values of 30.4 and 39.2 μM, respectively.

  14. Essential roles for calcium and calmodulin in G2/M progression in Aspergillus nidulans

    OpenAIRE

    1993-01-01

    nimT encodes a protein in Aspergillus nidulans that is required for tyrosine dephosphorylation of p34cdc2 and has a strong homology to cdc25-type proteins. Conditional mutation of nimT (nimT23 mutation) arrests cells in G2 at the restrictive temperature. After release of the temperature-sensitive nimT23 block, p34cdc2 undergoes tyrosine dephosphorylation and we showed that as cells entered mitosis, a rapid increase in calmodulin was observed. The increase in calmodulin and progression into mi...

  15. Terpenoids from the Marine-Derived Fungus Aspergillus fumigatus YK-7.

    Science.gov (United States)

    Wang, Yu; Li, Da-Hong; Li, Zhan-Lin; Sun, Yan-Jun; Hua, Hui-Ming; Liu, Tao; Bai, Jiao

    2015-12-28

    Two new β-bergamotane sesquiterpenoids, E-β-trans-5,8,11-trihydroxybergamot-9-ene (1) and β-trans-2β,5,15-trihydroxybergamot-10-ene (2), were isolated from the marine-derived fungus Aspergillus fumigatus YK-7, along with three known terpenoids 3-5. Their structures were determined by spectroscopic methods (1D and 2D NMR, HR-ESI-MS). Antiproliferative effects on human leukemic monocyte lymphoma U937 and human prostate cancer PC-3 cell lines were measured in vitro. Compound 4 exhibited potent activity against the U937 cell line with an IC50 value of 4.2 μM.

  16. Comparative genomics of citric-acid producing Aspergillus niger ATCC 1015 versus enzyme-producing CBS 513.88

    Energy Technology Data Exchange (ETDEWEB)

    Grigoriev, Igor V.; Baker, Scott E.; Andersen, Mikael R.; Salazar, Margarita P.; Schaap, Peter J.; Vondervoot, Peter J.I. van de; Culley, David; Thykaer, Jette; Frisvad, Jens C.; Nielsen, Kristen F.; Albang, Richard; Albermann, Kaj; Berka, Randy M.; Braus, Gerhard H.; Braus-Stromeyer, Susanna A.; Corrochano, Luis M.; Dai, Ziyu; Dijck, Piet W.M. van; Hofmann, Gerald; Lasure, Linda L.; Magnusson, Jon K.; Meijer, Susan L.; Nielsen, Jakob B.; Nielsen, Michael L.; Ooyen, Albert J.J. van; Panther, Kathyrn S.; Pel, Herman J.; Poulsen, Lars; Samson, Rob A.; Stam, Hen; Tsang, Adrian; Brink, Johannes M. van den; Atkins, Alex; Aerts, Andrea; Shapiro, Harris; Pangilinan, Jasmyn; Salamov, Asaf; Lou, Yigong; Lindquist, Erika; Lucas, Susan; Grimwood, Jane; Kubicek, Christian P.; Martinez, Diego; Peij, Noel N.M.E. van; Roubos, Johannes A.; Nielsen, Jens

    2011-04-28

    The filamentous fungus Aspergillus niger exhibits great diversity in its phenotype. It is found globally, both as marine and terrestrial strains, produces both organic acids and hydrolytic enzymes in high amounts, and some isolates exhibit pathogenicity. Although the genome of an industrial enzyme-producing A. niger strain (CBS 513.88) has already been sequenced, the versatility and diversity of this species compels additional exploration. We therefore undertook whole genome sequencing of the acidogenic A. niger wild type strain (ATCC 1015), and produced a genome sequence of very high quality. Only 15 gaps are present in the sequence and half the telomeric regions have been elucidated. Moreover, sequence information from ATCC 1015 was utilized to improve the genome sequence of CBS 513.88. Chromosome-level comparisons uncovered several genome rearrangements, deletions, a clear case of strain-specific horizontal gene transfer, and identification of 0.8 megabase of novel sequence. Single nucleotide polymorphisms per kilobase (SNPs/kb) between the two strains were found to be exceptionally high (average: 7.8, maximum: 160 SNPs/kb). High variation within the species was confirmed with exo-metabolite profiling and phylogenetics. Detailed lists of alleles were generated, and genotypic differences were observed to accumulate in metabolic pathways essential to acid production and protein synthesis. A transcriptome analysis revealed up-regulation of the electron transport chain, specifically the alternative oxidative pathway in ATCC 1015, while CBS 513.88 showed significant up-regulation of genes relevant to glucoamylase A production, such as tRNA-synthases and protein transporters. Our results and datasets from this integrative systems biology analysis resulted in a snapshot of fungal evolution and will support further optimization of cell factories based on filamentous fungi.[Supplemental materials (10 figures, three text documents and 16 tables) have been made available

  17. Effect of Vitronectin Bound to Insulin-Like Growth Factor-I and Insulin-Like Growth Factor Binding Protein-3 on Porcine Enamel Organ-Derived Epithelial Cells

    OpenAIRE

    Yoshinori Shinohara; Shuhei Tsuchiya; Kazuo Hatae; Honda, Masaki J

    2012-01-01

    The aim of this paper was to determine whether the interaction between IGF, IGFBP, and VN modulates the functions of porcine EOE cells. Enamel organs from 6-month-old porcine third molars were dissociated into single epithelial cells and subcultured on culture dishes pretreated with VN, IGF-I, and IGFBP-3 (IGF-IGFBP-VN complex). The subcultured EOE cells retained their capacity for ameloblast-related gene expression, as shown by semiquantitative reverse transcription-polymerase chain reaction...

  18. Aspergillus pragensis sp. nov. discovered during molecular reidentification of clinical isolates belonging to Aspergillus section Candidi.

    Science.gov (United States)

    Hubka, Vit; Lyskova, Pavlina; Frisvad, Jens C; Peterson, Stephen W; Skorepova, Magdalena; Kolarik, Miroslav

    2014-08-01

    The identity of nine clinical isolates recovered from Czech patients and presumptively identified as Aspergillus sp. section Candidi based on colony morphology was revised using sequences of β-tubulin, calmodulin gene sequence, and internal transcribed spacer rDNA. Six isolates were from suspected and proven onychomycosis, one from otitis externa, and two associated with probable invasive aspergillosis. The results showed that one Aspergillus candidus isolate was the cause of otitis externa, and both isolates obtained from sputa of patients with probable invasive aspergillosis were reidentified as A. carneus (sect. Terrei) and A. flavus (sect. Flavi). Three isolates from nail scrapings were identified as A. tritici, a verified agent of nondermatophyte onychomycosis. One isolate from toenail was determined to be A. candidus and the two isolates belonged to a hitherto undescribed species, Aspergillus pragensis sp. nov. This species is well supported by phylogenetic analysis based on β-tubulin and calmodulin gene and is distinguishable from other members of sect. Candidi by red-brown reverse on malt extract agar, slow growth on Czapek-Dox agar and inability to grow at 37°C. A secondary metabolite analysis was also provided with comparison of metabolite spectrum to other species. Section Candidi now encompasses five species for which a dichotomous key based on colony characteristics is provided. All clinical isolates were tested for susceptibilities to selected antifungal agents using the Etest and disc diffusion method. Overall sect. Candidi members are highly susceptible to common antifungals.

  19. A novel fungal fruiting structure formed by Aspergillus niger and Aspergillus carbonarius in grape berries.

    Science.gov (United States)

    Pisani, Cristina; Nguyen, Trang Thoaivan; Gubler, Walter Douglas

    2015-09-01

    Sour rot, is a pre-harvest disease that affects many grape varieties. Sour rot symptoms include initial berry cracking and breakdown of berry tissue. This is a disease complex with many filamentous fungi and bacteria involved, but is usually initiated by Aspergillus niger or Aspergillus carbonarius. Usually, by the time one sees the rot there are many other organisms involved and it is difficult to attribute the disease to one species. In this study two species of Aspergillus were shown to produce a previously unknown fruiting structure in infected berries. The nodulous morphology, bearing conidia, suggests them to be an 'everted polymorphic stroma'. This structure forms freely inside the berry pulp and assumes multiple shapes and sizes, sometimes sclerotium-like in form. It is composed of a mass of vegetative hyphae with or without tissue of the host containing spores or fruiting bodies bearing spores. Artificially inoculated berries placed in soil in winter showed the possible overwintering function of the fruiting body. Inoculated berry clusters on standing vines produced fruiting structures within 21 d post inoculation when wounds were made at veraison or after (July-September). Histological studies confirmed that the fruiting structure was indeed fungal tissue.

  20. Azole drug import into the pathogenic fungus Aspergillus fumigatus.

    Science.gov (United States)

    Esquivel, Brooke D; Smith, Adam R; Zavrel, Martin; White, Theodore C

    2015-01-01

    The fungal pathogen Aspergillus fumigatus causes serious illness and often death when it invades tissues, especially in immunocompromised individuals. The azole class of drugs is the most commonly prescribed treatment for many fungal infections and acts on the ergosterol biosynthesis pathway. One common mechanism of acquired azole drug resistance in fungi is the prevention of drug accumulation to toxic levels in the cell. While drug efflux is a well-known resistance strategy, reduced azole import would be another strategy to maintain low intracellular azole levels. Recently, azole uptake in Candida albicans and other yeasts was analyzed using [(3)H]fluconazole. Defective drug import was suggested to be a potential mechanism of drug resistance in several pathogenic fungi, including Cryptococcus neoformans, Candida krusei, and Saccharomyces cerevisiae. We have adapted and developed an assay to measure azole accumulation in A. fumigatus using radioactively labeled azole drugs, based on previous work done with C. albicans. We used this assay to study the differences in azole uptake in A. fumigatus isolates under a variety of drug treatment conditions, with different morphologies and with a select mutant strain with deficiencies in the sterol uptake and biosynthesis pathway. We conclude that azole drugs are specifically selected and imported into the fungal cell by a pH- and ATP-independent facilitated diffusion mechanism, not by passive diffusion. This method of drug transport is likely to be conserved across most fungal species.

  1. Pulmonary Immune Responses to Aspergillus Fumigatus in Rats

    Institute of Scientific and Technical Information of China (English)

    MIRKOV Ivana; EL-MUZGHI Amal Atia Mhfuod; DJOKIC Jelena; NINKOV Marina; POPOV ALEKSANDROV Aleksandra; GLAMOCLIJA Jasmina; KATARANOVSKI Milena

    2014-01-01

    Objective To evaluate immunologic mechanisms underlying Aspergillus fumigatus pulmonary infections in immunocompetent Dark Agouti (DA) and Albino Oxford (AO) rats recognized as being susceptible to some inflammatory diseases in different manners. Methods Lung fungal burden (quantitative colony forming units, CFU, assay), leukocyte infiltration (histology, cell composition) and their function (phagocytosis, oxidative activity, CD11b adhesion molecule expression) and cytokine interferon-γ(IFN-γ) and interleukin-17 and-4 (IL-17 and IL-4) lung content were evaluated following infection (intratracheally, 1x107 conidia). Results Slower reduction of fungal burden was observed in AO rats in comparison with that in DA rats, which was coincided with less intense histologically evident lung cell infiltration and leukocyte recovery as well as lower level of most of the their activities including intracellular myeloperoxidase activity, the capacity of nitroblue tetrazolium salt reduction and CD11b adhesion molecule expression (except for phagocytosis of conidia) in these rats. Differential patterns of changes in proinflammatory cytokine levels (unchanged levels of IFN-γand transient increase of IL-17 in AO rats vs continuous increase of both cytokines in DA rats) and unchanged levels of IL-4 were observed. Conclusion Genetically-based differences in the pattern of antifungal lung leukocyte activities and cytokine milieu, associated with differential efficiency of fungal elimination might be useful in the future use of rat models in studies of pulmonary aspergillosis.

  2. Hide, keep quiet, and keep low: properties that make Aspergillus fumigatus a successful lung pathogen

    Directory of Open Access Journals (Sweden)

    Natalia eEscobar

    2016-04-01

    Full Text Available Representatives of the genus Aspergillus are opportunistic fungal pathogens. Their conidia can reach the alveoli by inhalation and can give rise to infections in immunocompromised individuals. Aspergillus fumigatus is the causal agent of invasive aspergillosis in nearly 90 % of the cases. It is not yet well established what makes this fungus more pathogenic than other aspergilli such as Aspergillus niger. Here, we show that A. fumigatus and A. niger conidia adhere with similar efficiency to lung epithelial A549 cells but A. fumigatus conidia internalized 17% more efficiently. Conidia of both aspergilli were taken up in phagolysosomes 8 h after the challenge. These organelles only acidified in the case of A. niger, which is probably due to the type of melanin coating of the conidia. Viability of both types of conidia was not affected after uptake in the phagolysosomes. Germination of A. fumigatus and A. niger conidia in the presence of epithelial cells was delayed when compared to conidia in the medium. However, germination of A. niger conidia was still higher than that of A. fumigatus 10 h after exposure to A549 cells. Remarkably, A. fumigatus hyphae grew mainly parallel to the epithelium, while growth direction of A. niger hyphae was predominantly perpendicular to the plane of the cells. Neutrophils reduced germination and hyphal growth of A. niger, but not of A fumigatus, in presence of epithelial cells. Taken together, efficient internalization, delayed germination, and hyphal growth parallel to the epithelium gives a new insight into what could be the causes for the success of A. fumigatus compared to A. niger as an opportunistic pathogen in the lung.

  3. The Aspergillus Genome Database, a curated comparative genomics resource for gene, protein and sequence information for the Aspergillus research community.

    Science.gov (United States)

    Arnaud, Martha B; Chibucos, Marcus C; Costanzo, Maria C; Crabtree, Jonathan; Inglis, Diane O; Lotia, Adil; Orvis, Joshua; Shah, Prachi; Skrzypek, Marek S; Binkley, Gail; Miyasato, Stuart R; Wortman, Jennifer R; Sherlock, Gavin

    2010-01-01

    The Aspergillus Genome Database (AspGD) is an online genomics resource for researchers studying the genetics and molecular biology of the Aspergilli. AspGD combines high-quality manual curation of the experimental scientific literature examining the genetics and molecular biology of Aspergilli, cutting-edge comparative genomics approaches to iteratively refine and improve structural gene annotations across multiple Aspergillus species, and web-based research tools for accessing and exploring the data. All of these data are freely available at http://www.aspgd.org. We welcome feedback from users and the research community at aspergillus-curator@genome.stanford.edu.

  4. Characterization and biological treatment of colored textile wastewaters from the typical Tunisian hat Chechia dyeing using newly isolated Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Hajer Barouni

    2016-09-01

    Full Text Available This study aimed to characterize and investigate, for the first time, the treatment of real colored wastewaters from the artisanal dyeing of the typical Tunisian hat Chechia, using a newly isolated fungal strain. This textile effluent was a mixture called Mix of colored wastewaters from the three main types of Chechia. The major pollutant of the Mix was the toxic Azo dye Amaranth Acid or Acid Red 27. The fungal strain that made the cleanup was discovered in a Chechia dyeing wastewater’s container and identified by ITS rDNA gene sequencing. This isolated Aspergillus niger showed interesting performances on the demonstration of Chechia wastewater’s biodegradation in batch cultures. In order to understand the effect of agitation, Mix dilution and inoculum size on decolourisation and pollution removal, a full factorial experimental design 23 was set up. At the optimal conditions which were 20% inoculum size, 25% Chechia Mix dilution and an agitation of 100 rpm, Aspergillus niger was able to remove color as high as 70.18±2.84% at an initial dye concentration of 1346.6±0.01 mg/L, and to reduce COD to 74.17±14.52% at an initial COD of 4157±422 mg/L. FT-IR spectra analysis confirmed the decolourisation by biodegradation and transformation of the dyes. The treatment by the isolated Aspergillus niger could be successfully applied as a sustainable method to solve one of handicraft dyeing plants environmental management issues.

  5. Systems Factorial Technology Explained to Humans

    Directory of Open Access Journals (Sweden)

    Harding, Bradley

    2016-01-01

    Full Text Available The study of mental processes is at the forefront of research in cognitive psychology. However, the ability to identify the architectures responsible for specific behaviors is often quite difficult. To alleviate this difficulty, recent progress in mathematical psychology has brought forth Systems Factorial Technology (SFT; Townsend & Nozawa, 1995. Encompassing a series of analyses, SFT can diagnose and discriminate between five types of information processing architectures that possibly underlie a mental process. Despite the fact that SFT has led to new discoveries in cognitive psychology, the methodology itself remains far from intuitive to newcomers. This article therefore seeks to provide readers with a simple tutorial and a rudimentary introduction to SFT. This tutorial aims to encourage newcomers to read more about SFT and also to add it to their repertoire of analyses.

  6. Future e+ e- Flavor Factories: accelerator challenges

    CERN Document Server

    Biagini, M E

    2008-01-01

    Operation of the B-Factories (PEP-II and KEKB) has been very successful, both having exceeded their design peak and integrated luminosity and provided a huge amount of good data to the experiments. Proposal for upgrades, in order to achieve about two order of magnitude larger luminosity, are in progress in Japan, with Super-KEKB, and in Europe, with SuperB. Very high beam intensity, very short bunch length and low Interaction Point beta-functions are the key points of the Japanese design, very challenging for the hardware components (RF, vacuum). On the other hand SuperB exploits a new collision scheme, namely large Piwinski angle and "crab waist", which will allow to reach a luminosity two order of magnitude larger without increasing beam currents and decreasing bunch lengths. In this talk the present status of the two projects will be reviewed.

  7. Producción escrita: Estructura factorial

    Directory of Open Access Journals (Sweden)

    María Esperanza HERRERA GARCÍA

    2009-11-01

    Full Text Available Este estudio se planteó con la idea de comprobar de forma empírica si los indicadores lingüísticos obtenidos en investigaciones realizadas sobre comprensión lectora aparecían también como significativos en el análisis de la producción escrita. Si bien la investigación psicológica ha demostrado en numerosas ocasiones la relación existente entre comprensión y producción en el sentido de una maduración anterior de la primera con respecto a la segunda, podemos considerar a priori la existencia de indicadores que nos permitan medir ambos aspectos. Para ello buscamos la estructura factorial que presentan los indicadores lingüísticos que vamos a considerar y comprobaremos si presenta los factores encontrados en comprensión lectora.

  8. Moscow Meson Factory DTL RF System Upgrade

    CERN Document Server

    Esin, S K; Kvasha, A I; Serov, V L

    2004-01-01

    The last paper devoted to description of the first part (DTL) RF system of Moscow Meson Factory upgrade was published in the Proceedings of PAC95 Conference in Dallas. Since then some new works directed at improvement of reliability and efficiency of the RF system were carried out. Among them there are a new powerful pulse triode “Katran” installed in the output RF power amplifiers (PA) of three channels, modifications of the anode modulator control circuit and crow-bar system, a new additional RF channel for RF supply of RFQ and some alterations in placing of the anode modulator equipment decreasing a level of interference’s at crow-bar circuits. Some new checked at MMF RF channels ideas concerning of PA tuning are of interest for people working in this sphere of activity.

  9. AutoPyFactory and the Cloud

    CERN Document Server

    Caballero, J; The ATLAS collaboration; Love, P

    2013-01-01

    AutoPyFactory (APF) is a next-generation pilot submission framework that has been used as part of the ATLAS workload management system (PANDA) for two years. APF is reliable, scalable, and offers easy and flexible configuration. Using a plugin-based architecture, APF polls for information from configured information and batch systems (including grid sites), decides how many additional pilot jobs are needed, and submits them. With the advent of cloud computing, providing resources goes beyond submitting pilots to grid sites. Now, the resources on which the pilot will run also need to be managed. Handling both pilot submission and controlling the virtual machine life cycle (creation, retirement, and termination) from the same framework allows robust and efficient management of the process. In this paper we describe the design and implementation of these virtual machine management capabilities of APF. Expanding on our plugin-based approach, we allow cascades of virtual resources associated with a job queue. A si...

  10. High-gravity brewing utilizing factorial design

    Directory of Open Access Journals (Sweden)

    R. B. Almeida

    2000-06-01

    Full Text Available A number of factors can influence the behavior of yeast during fermentation. Some of these factors (initial wort concentration, initial pH and percentage of corn syrup in the composition of the wort were studied in order to determine their influence on the productivity of fermentation. Fermentations were carried out at 25ºC utilizing a 2³ factorial design of these factors. The results showed that the percentage of corn syrup had no influence on process productivity, whereas initial pH and especially initial wort concentration did. It can be concluded that using pH and initial wort concentration values higher than those utilized in this work (5.5 and 20ºP, respectively will result in a higher productivity.

  11. Production of amylases by Aspergillus tamarii

    Directory of Open Access Journals (Sweden)

    Moreira Fabiana Guillen

    1999-01-01

    Full Text Available A strain of Aspergillus tamarii, a filamentous fungus isolated from soil, was able to produce both a-amylase and glucoamylase activities in mineral media supplemented with 1% (w/v starch or maltose as the carbon source. Static cultivation led to significantly higher yields than those obtained using shaking culture. The production of amylases was tolerant to a wide range of initial culture pH values (from 4 to 10 and temperature (from 25 to 42oC. Two amylases, one a-amylase and one glucoamylase, were separated by ion exchange chromatography. Both partially purified enzymes had optimal activities at pH values between 4.5 and 6.0 and were stable under acid conditions (pH 4.0-7.0. The enzymes exhibited optimal activities at temperatures between 50o and 60o C and were stable for more than ten hours at 55oC.

  12. Environmental fungicides and triazole resistance in Aspergillus.

    Science.gov (United States)

    Bowyer, Paul; Denning, David W

    2014-02-01

    Fungal diseases are problematic in both human health and agriculture. Treatment options are limited and resistance may emerge. The relatively recent recognition of triazole resistance in Aspergillus fumigatus has prompted questioning of the origin of resistance. While multiple mechanisms are described in clinical isolates from triazole-treated patients, some de novo resistance is also recognised, especially attributable to TR34 /L98H. Such strains probably arose in the environment, and, indeed, multiple studies have now demonstrated TR(34) /L98H triazole resistance strains of A. fumigatus from soil. Docking and other in vitro studies are consistent with environmental resistance induction through exposure to certain triazole fungicides, notably difenoconazole, propiconazole, epoxiconazole, bromuconazole and tebuconazole. This article addresses the potential implications of this issue for both human health and food security.

  13. Phenylquinolinones with antitumor activity from the Indian Ocean-derived fungus Aspergillus versicolor Y31-2

    Science.gov (United States)

    Li, Peihai; Fan, Yaqin; Chen, Hao; Chao, Yaxi; Du, Ning; Chen, Junhui

    2016-09-01

    Two phenylquinolinones, including one new compound ( 1) and a previously isolated compound ( 2), were isolated from the ethyl acetate extracts of the fungus Aspergillus versicolor Y31-2, which was obtained from seawater samples collected from the Indian Ocean. The structures of these compounds were established by spectroscopic analyses. 4-(3-Hydroxyphenyl)-3-methoxyquinolin-2(1H)-one ( 1) exhibited moderate cytotoxicity against MCF-7 (human breast carcinoma cell line) and SMMC-7721 (human liver cancer cell line) cells with IC50 values of 16.6 and 18.2 μmol/L, respectively. To the best of our knowledge, this study represents the first reported account of the isolation of compounds 1 and 2 as the secondary metabolites of the seawater derived fungus Aspergillus versicolor from the Indian Ocean.

  14. Three new species of Aspergillus from Amazonian forest soil (Ecuador).

    Science.gov (United States)

    Mares, Donatella; Andreotti, Elisa; Maldonado, Maria Elena; Pedrini, Paola; Colalongo, Chiara; Romagnoli, Carlo

    2008-09-01

    From an undisturbed natural forest soil in Ecuador, three fungal strains of the genus Aspergillus were isolated. Based on molecular and morphological features they are described as three new species, named A. quitensis, A. amazonicus, and A. ecuadorensis.

  15. Septic arthritis due to tubercular and Aspergillus co-infection.

    Science.gov (United States)

    Kumar, Mukesh; Thilak, Jai; Zahoor, Adnan; Jyothi, Arun

    2016-01-01

    Aspergillus septic arthritis is a rare and serious medical and surgical problem. It occurs mainly in immunocompromised patients. Aspergillus fumigatus is the most common causative organism followed by Aspergillus flavus. The most common site affected is knee followed by shoulder, ankle, wrist, hip and sacroiliac joint. Debridement and voriconazole are primary treatment of articular aspergilosis. To the best of our knowledge, there are no reported cases of co-infection of tuberculosis (TB) and Aspergillus infecting joints. We report a case of co-infection of TB and A. flavus of hip and knee of a 60-year-old male, with type 2 diabetes mellitus. He was treated with debridement, intravenous voriconazole, and antitubercular drugs.

  16. Reconstruction of the central carbon metabolism of Aspergillus niger

    DEFF Research Database (Denmark)

    David, Helga; Åkesson, Mats Fredrik; Nielsen, Jens

    2003-01-01

    The topology of central carbon metabolism of Aspergillus niger was identified and the metabolic network reconstructed, by integrating genomic, biochemical and physiological information available for this microorganism and other related fungi. The reconstructed network may serve as a valuable...

  17. An amendment of Aspergillus section Candidi based on chemotaxonomical evidence

    DEFF Research Database (Denmark)

    Rahbaek, L.; Frisvad, Jens Christian; Christophersen, C.

    2000-01-01

    A novel 2,2 '-epoxy-terphenyllin, candidusin C, in addition to the well known secondary metabolites terphenyllin, 3-hydroxyterpenyllin and chlorflavonin, has been isolated from the chemically unexplored fungus Aspergillus campestris. The latter three are known secondary metabolites from Aspergillus...... candidus and therefore a large number of Aspergilli were screened for production of these compounds to see whether they could be regarded as chemotaxonomical indicators of section membership in the monotypic Aspergillus section Candidi. The results indicated that A. campestris and A. taichungensis should...... be placed in Candidi and this was further confirmed by morphological and physiological similarities. Three species outside the section Candidi produced candidusin related secondary metabolites: Aspergillus arenarius, A. ellipticus and Penicillium raistrickii. Chlorflavonin, however, was only found...

  18. Biodiversity of Aspergillus species in some important agricultural products

    DEFF Research Database (Denmark)

    Perrone, Giancarlo; Susca, A.,; Cozzi, G.

    2007-01-01

    The genus Aspergillus is one of the most important filamentous fungal genera. Aspergillus species are used in the fermentation industry, but they are also responsible of various plant and food secondary rot, with the consequence of possible accumulation of mycotoxins. The aflatoxin producing A....... flavus and A. parasiticus, and ochratoxinogenic A. niger, A. ochraceus and A. carbonarius species are frequently encountered in agricultural products. Studies on the biodiversity of toxigenic Aspergillus species is useful to clarify molecular, ecological and biochemical characteristics of the different...... occurring in agricultural fields. Altogether nine different black Aspergillus species can be found on grapes which are often difficult to identify with classical methods. The polyphasic approach used in our studies led to the identification of three new species occurring on grapes: A. brasiliensis, A...

  19. Septic arthritis due to tubercular and Aspergillus co-infection

    Directory of Open Access Journals (Sweden)

    Mukesh Kumar

    2016-01-01

    Full Text Available Aspergillus septic arthritis is a rare and serious medical and surgical problem. It occurs mainly in immunocompromised patients. Aspergillus fumigatus is the most common causative organism followed by Aspergillus flavus. The most common site affected is knee followed by shoulder, ankle, wrist, hip and sacroiliac joint. Debridement and voriconazole are primary treatment of articular aspergilosis. To the best of our knowledge, there are no reported cases of co-infection of tuberculosis (TB and Aspergillus infecting joints. We report a case of co-infection of TB and A. flavus of hip and knee of a 60-year-old male, with type 2 diabetes mellitus. He was treated with debridement, intravenous voriconazole, and antitubercular drugs.

  20. Extracellular siderophores of rapidly growing Aspergillus nidulans and Penicillium chrysogenum

    OpenAIRE

    Charlang, G; Horowitz, R M; Lowy, P H; Ng, B.; Poling, S M; Horowitz, N. H.

    1982-01-01

    The highly active extracellular siderophores previously detected in young cultures of Aspergillus nidulans and Penicillium chrysogenum have been identified as the cyclic ester fusigen (fusarinine C), and its open-chain form, fusigen B (fusarinine B).

  1. Aerobiological analysis in a salami factory: a possible case of extrinsic allergic alveolitis by Penicillium camembertii.

    Science.gov (United States)

    Marchisio, V F; Sulotto, F; Botta, G C; Chiesa, A; Airaudi, D; Anastasi, A

    1999-08-01

    A 39-year-old man was hospitalized with a history of fatigue, dyspnoea and low grade fever which seemed to be related to his working environment. The patient was employed in a salami factory, working near the area where the salami are seasoned with fungal inocula. Chest X-ray showed diffuse initial changes of reticulonodular pattern that disappeared after a brief course of steroids therapy. Precipitating antibodies to Penicillium notatum and Aspergillus fumigatus were found both in plasma and bronchoalveolar lavage fluid. This, together with the finding of a lymphocytic alveolitis with CD4+ depletion and CD8+ increase, suggested the possibility of extrinsic allergic alveolitis of fungal aetiology. Qualitative and quantitative monitoring with an impinger of both the working and outside environment for aerial fungal concentration demonstrated a very high level of contamination (up to 1.14x10(9) fungal propagules m-3 of air) and an inside/outside ratio from 21 to about 2000. Penicillium camembertii was the most common species found in all the indoor sites (60-100% of the fungal load). The patient's BALF and serum both displayed precipitating antibodies to P. camembertii from the powder used for the inoculum and the air samples. These results together with the patient's working history gave some evidence of relationship between the indoor P. camembertii concentration and the patient's symptoms.

  2. Susceptibility breakpoints for amphotericin B and Aspergillus species in an in vitro pharmacokinetic-pharmacodynamic model simulating free-drug concentrations in human serum

    NARCIS (Netherlands)

    Elefanti, A.; Mouton, J.W.; Verweij, P.E.; Zerva, L.; Meletiadis, J.

    2014-01-01

    Although conventional amphotericin B was for many years the drug of choice and remains an important agent against invasive aspergillosis, reliable susceptibility breakpoints are lacking. Three clinical Aspergillus isolates (Aspergillus fumigatus, Aspergillus flavus, and Aspergillus terreus) were tes

  3. Polyphasic taxonomy of Aspergillus fumigatus and related species

    DEFF Research Database (Denmark)

    Hong, S.B.; Go, S.J.; Shin, H.D.

    2005-01-01

    The variability within Aspergillus fumigalus Fresenius and related species was examined using macro-, micro-morphology, growth temperature regimes and extrolite patterns. In addition, DNA analyses including partial beta-tubulin, calmodulin and actin gene sequences were used. Detailed examination....... fumigatus and the other Aspergillus section Fumigati species, including the teleomorph Neosartorya, are proposed as two new species. A. fumigatiaffinis spec. nov. produces the extrolites auranthine, cycloechinulin, helvolic acid, neosartorin, palitantin, pyripyropens, tryptoquivalins and tryptoquivalons...

  4. GridFactory - Distributed computing on ephemeral resources

    DEFF Research Database (Denmark)

    Orellana, Frederik; Niinimaki, Marko

    2011-01-01

    on demand, across organizational boundaries. Other features include firewall-friendliness, integrated software provisioning and virtual machine management. We describe the design and architecture of Grid Factory and four applications that utilize it. Moreover, we compare the performance of Grid Factory...

  5. Factory Gate Pricing : An Analysis of the Dutch Retail Distribution

    NARCIS (Netherlands)

    Le Blanc, H.M.; Cruijssen, F.C.A.M.; Fleuren, H.A.; de Koster, M.B.M.

    2004-01-01

    Factory Gate Pricing (FGP) is a relatively new phenomenon in retail distribution.Under FGP, products are no longer delivered at the retailer distribution center, but collected by the retailer at the factory gates of the suppliers.Owing to both the asymmetry in the distribution networks (the supplier

  6. Factory Gate Pricing: An Analysis of the Dutch Retail Distribution

    NARCIS (Netherlands)

    H.M. le Blanc; F. Cruijssen (Frans); H.A. Fleuren; M.B.M. de Koster (René)

    2004-01-01

    textabstractFactory Gate Pricing (FGP) is a relatively new phenomenon in retail distribution. Under FGP, products are no longer delivered at the retailer distribution center, but collected by the retailer at the factory gates of the suppliers. Owing to both the asymmetry in the distribution networks

  7. China’s Largest Copper Foil Factory Opened

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    <正>On October 28,civil engineering and steel structure projects by Nanya No.3 Copper Foil Factory,the largest copper foil factory on the Chinese mainland by Shanghai Baoye Group Corp.,Ltd.,opened on Changjiang Nan Road in Kunshan’s

  8. Students Tackle Academics in Practical Context at Skateboard Factory

    Science.gov (United States)

    Pearson, George

    2012-01-01

    High school student Nick Robertson came to Oasis Skateboard Factory (OSF) from a suburban Toronto district. He was surfing the web when he spotted a reference to Oasis Skateboard Factory, an alternative program in the Toronto District School Board. He says his first reaction was "Skateboards in school? It didn't seem possible." He…

  9. Connection Among Some Optimal Criteria for Symmetrical Fractional Factorial Designs

    Institute of Scientific and Technical Information of China (English)

    Hong Qin; Ming-yao Ai; Jian-hui Ning

    2005-01-01

    A fundamental and practical question for fractional factorial designs is the issue of optimal factor assignment. Recently, some new criteria, such as generalized minimum aberration, WV-criterion, NB-criterion and uniformity criterion are proposed for comparing and selecting fractions. In this paper, we indicate that these criteria agree quite well for symmetrical fraction factorial designs.

  10. 27 CFR 40.70 - Separation of and access to factory.

    Science.gov (United States)

    2010-04-01

    ... to factory. 40.70 Section 40.70 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND... § 40.70 Separation of and access to factory. Where the factory consists of a portion of a building, or where portions of buildings are part of the factory, the factory shall be completely separated by...

  11. Hypersensitivity testing for Aspergillus fumigatus IgE is significantly more sensitive than testing for Aspergillus niger IgE.

    Science.gov (United States)

    Selvaggi, Thomas A; Walco, Jeremy P; Parikh, Sujal; Walco, Gary A

    2012-02-01

    We sought to determine if sufficient redundancy exists between specific IgE testing for Aspergillus fumigatus and Aspergillus niger to eliminate one of the assays in determining Aspergillus hypersensitivity. We reviewed regional laboratory results comparing A fumigatus-specific IgE with A niger-specific IgE using the Pharmacia UniCAP system (Pharmacia, Kalamazoo, MI). By using the Fisher exact test as an index of concordance among paired results, we showed a significant difference between 109 paired samples for the presence of specific IgE to A fumigatus and A niger (P niger; no specimen was positive for A niger and negative for A fumigatus. We conclude that A fumigatus-specific IgE is sufficient to detect Aspergillus hypersensitivity. The assay for A niger-specific IgE is redundant, less sensitive, and unnecessary if the assay for specific IgE for A fumigatus is performed.

  12. The Three-dimensional Digital Factory for Shipbuilding Technology Research

    Directory of Open Access Journals (Sweden)

    Xu Wei

    2016-01-01

    Full Text Available The three-dimensional digital factory technology research is the hotspot in shipbuilding recently. The three-dimensional digital factory technology not only focus on design the components of the product, but also discuss on the simulation and analyses of the production process.Based on the three-dimensional model, the basic data layer, application control layer and the presentation layer of hierarchical structure are established in the three-dimensional digital factory of shipbuilding in this paper. And the key technologies of three-dimensional digital factory of shipbuilding are analysed. Finally, a case study is applied and the results show that the three-dimensional digital factory will play an important role in the future.

  13. Green Tea Epigallocatechin Gallate Exhibits Anticancer Effect in Human Pancreatic Carcinoma Cells via the Inhibition of Both Focal Adhesion Kinase and Insulin-Like Growth Factor-I Receptor

    Directory of Open Access Journals (Sweden)

    Hoang Anh Vu

    2010-01-01

    Full Text Available The exact molecular mechanism by which epigallocatechin gallate (EGCG suppresses human pancreatic cancer cell proliferation is unclear. We show here that EGCG-treated pancreatic cancer cells AsPC-1 and BxPC-3 decrease cell adhesion ability on micro-pattern dots, accompanied by dephosphorylations of both focal adhesion kinase (FAK and insulin-like growth factor-1 receptor (IGF-1R whereas retained the activations of mitogen-activated protein kinase and mammalian target of rapamycin. The growth of AsPC-1 and BxPC-3 cells can be significantly suppressed by EGCG treatment alone in a dose-dependent manner. At a dose of 100 μM which completely abolishes activations of FAK and IGF-1R, EGCG suppresses more than 50% of cell proliferation without evidence of apoptosis analyzed by PARP cleavage. Finally, the MEK1/2 inhibitor U0126 enhances growth-suppressive effect of EGCG. Our data suggests that blocking FAK and IGF-1R by EGCG could prove valuable for targeted therapy, which can be used in combination with other therapies, for pancreatic cancer.

  14. From Neanderthal to nanobiotech: from plant potions to pharming with plant factories.

    Science.gov (United States)

    Sourrouille, Christophe; Marshall, Brian; Liénard, David; Faye, Loïc

    2009-01-01

    Plants were the main source for human drugs until the beginning of the nineteenth century when plant-derived pharmaceuticals were partly supplanted by drugs produced by the industrial methods of chemical synthesis. During the last decades of the twentieth century, genetic engineering has offered an alternative to chemical synthesis, using bacteria, yeasts and animal cells as factories for the production of therapeutic proteins. After a temporary decrease in interest, plants are rapidly moving back into human pharmacopoeia, with the recent development of plant-based recombinant protein production systems offering a safe and extremely cost-effective alternative to microbial and mammalian cell cultures. In this short review, we will illustrate that current improvements in plant expression systems are making them suitable as alternative factories for the production of either simple or highly complex therapeutic proteins.

  15. Initial intracellular proteome profile of Aspergillus niger biofilms

    Directory of Open Access Journals (Sweden)

    Gretty K. Villena

    2011-07-01

    Full Text Available An initial profiling of the intracellular proteome of Aspergillus niger ATCC 10864 biofilm cultures developed on polyester cloth was carried out by using 2D-PAGE and MS-TOF analysis and it was compared to the proteome of conventionally grown free-living submerged cultures. A number of 2D-PAGE protein spots from both types of cultures were subjected to MS-TOF analysis and data interrogation of the NCBI nr database available for this species. Proteomic maps showed different expression patterns in both culture systems with differentially expressed proteins in each case. In biofilm cultures, 19% and 32% of the selected protein spots were over- expressed and differentially expressed, respectively. On the contrary, in free-living cultures, 44% and 7% of the selected protein spots were over-expressed and differentially expressed, respectively. Although preliminary, results presented in this paper show that there are significant differences between the proteomes of A. niger biofilm and free-living mycelia. It seems that cell adhesion is the most important stimulus responsible for biofilm development which is the basis of Surface Adhesion Fermentation.

  16. Validation of a new Aspergillus real-time PCR assay for direct detection of Aspergillus and azole resistance of Aspergillus fumigatus on bronchoalveolar lavage fluid.

    Science.gov (United States)

    Chong, Ga-Lai M; van de Sande, Wendy W J; Dingemans, Gijs J H; Gaajetaan, Giel R; Vonk, Alieke G; Hayette, Marie-Pierre; van Tegelen, Dennis W E; Simons, Guus F M; Rijnders, Bart J A

    2015-03-01

    Azole resistance in Aspergillus fumigatus is increasingly reported. Here, we describe the validation of the AsperGenius, a new multiplex real-time PCR assay consisting of two multiplex real-time PCRs, one that identifies the clinically relevant Aspergillus species, and one that detects the TR34, L98H, T289A, and Y121F mutations in CYP51A and differentiates susceptible from resistant A. fumigatus strains. The diagnostic performance of the AsperGenius assay was tested on 37 bronchoalveolar lavage (BAL) fluid samples from hematology patients and 40 BAL fluid samples from intensive care unit (ICU) patients using a BAL fluid galactomannan level of ≥1.0 or positive culture as the gold standard for detecting the presence of Aspergillus. In the hematology and ICU groups combined, there were 22 BAL fluid samples from patients with invasive aspergillosis (IA) (2 proven, 9 probable, and 11 nonclassifiable). Nineteen of the 22 BAL fluid samples were positive, according to the gold standard. The optimal cycle threshold value for the presence of Aspergillus was Aspergillus species and 14 A. fumigatus samples). This resulted in a sensitivity, specificity, and positive and negative predictive values of 88.9%, 89.3%, 72.7%, and 96.2%, respectively, for the hematology group and 80.0%, 93.3%, 80.0%, and 93.3%, respectively, in the ICU group. The CYP51A real-time PCR confirmed 12 wild-type and 2 resistant strains (1 TR34-L98H and 1 TR46-Y121F-T289A mutant). Voriconazole therapy failed for both patients. The AsperGenius multiplex real-time PCR assay allows for sensitive and fast detection of Aspergillus species directly from BAL fluid samples. More importantly, this assay detects and differentiates wild-type from resistant strains, even if BAL fluid cultures remain negative.

  17. Production of Polygalacturonases by Aspergillus section Nigri Strains in a Fixed Bed Reactor

    Directory of Open Access Journals (Sweden)

    Marília Maciel

    2013-01-01

    Full Text Available Polygalacturonases (PG are pectinolytic enzymes that have technological, functional and biological applications in food processing, fruit ripening and plant-fungus interactions, respectively. In the present, a microtitre plate methodology was used for rapid screening of 61 isolates of fungi from Aspergillus section Nigri to assess production of endo- and exo-PG. Studies of scale-up were carried out in a fixed bed reactor operated under different parameters using the best producer strain immobilised in orange peels. Four experiments were conducted under the following conditions: the immobilised cells without aeration; immobilised cells with aeration; immobilised cells with aeration and added pectin; and free cells with aeration. The fermentation was performed for 168 h with removal of sample every 24 h. Aspergillus niger strain URM 5162 showed the highest PG production. The results obtained indicated that the maximum endo- and exo-PG activities (1.18 U·mL−1 and 4.11 U·mL−1, respectively were obtained when the reactor was operating without aeration. The microtitre plate method is a simple way to screen fungal isolates for PG activity detection. The fixed bed reactor with orange peel support and using A. niger URM 5162 is a promising process for PG production at the industrial level.

  18. Production of polygalacturonases by Aspergillus section Nigri strains in a fixed bed reactor.

    Science.gov (United States)

    Maciel, Marília; Ottoni, Cristiane; Santos, Cledir; Lima, Nelson; Moreira, Keila; Souza-Motta, Cristina

    2013-01-28

    Polygalacturonases (PG) are pectinolytic enzymes that have technological, functional and biological applications in food processing, fruit ripening and plant-fungus interactions, respectively. In the present, a microtitre plate methodology was used for rapid screening of 61 isolates of fungi from Aspergillus section Nigri to assess production of endo- and exo-PG. Studies of scale-up were carried out in a fixed bed reactor operated under different parameters using the best producer strain immobilised in orange peels. Four experiments were conducted under the following conditions: the immobilised cells without aeration; immobilised cells with aeration; immobilised cells with aeration and added pectin; and free cells with aeration. The fermentation was performed for 168 h with removal of sample every 24 h. Aspergillus niger strain URM 5162 showed the highest PG production. The results obtained indicated that the maximum endo- and exo-PG activities (1.18 U · mL-1 and 4.11 U · mL-1, respectively) were obtained when the reactor was operating without aeration. The microtitre plate method is a simple way to screen fungal isolates for PG activity detection. The fixed bed reactor with orange peel support and using A. niger URM 5162 is a promising process for PG production at the industrial level.

  19. A review on Lactococcus lactis: from food to factory.

    Science.gov (United States)

    Song, Adelene Ai-Lian; In, Lionel L A; Lim, Swee Hua Erin; Rahim, Raha Abdul

    2017-04-04

    Lactococcus lactis has progressed a long way since its discovery and initial use in dairy product fermentation, to its present biotechnological applications in genetic engineering for the production of various recombinant proteins and metabolites that transcends the heterologous species barrier. Key desirable features of this gram-positive lactic acid non-colonizing gut bacteria include its generally recognized as safe (GRAS) status, probiotic properties, the absence of inclusion bodies and endotoxins, surface display and extracellular secretion technology, and a diverse selection of cloning and inducible expression vectors. This have made L. lactis a desirable and promising host on par with other well established model bacterial or yeast systems such as Escherichia coli, Salmonella cerevisiae and Bacillus subtilis. In this article, we review recent technological advancements, challenges, future prospects and current diversified examples on the use of L. lactis as a microbial cell factory. Additionally, we will also highlight latest medical-based applications involving whole-cell L. lactis as a live delivery vector for the administration of therapeutics against both communicable and non-communicable diseases.

  20. Does fungicide application in vineyards induce resistance to medical azoles in Aspergillus species?

    Science.gov (United States)

    Lago, Magali; Aguiar, Ana; Natário, André; Fernandes, Carla; Faria, Miguel; Pinto, Eugénia

    2014-09-01

    This study assessed if the use of sterol demethylase inhibitor fungicides in vineyard production can induce resistance to azoles in Aspergillus strains and if it can induce selection of resistant species. We also tried to identify the Aspergillus species most prevalent in the vineyards. Two vineyards from northern Portugal were selected from "Vinhos Verdes" and "Douro" regions. The vineyards were divided into plots that were treated or not with penconazole (PEN). In each vineyard, air, soil, and plant samples were collected at three different times. The strains of Aspergillus spp. were isolated and identified by morphological and molecular techniques. We identified 46 Aspergillus section Nigri, eight Aspergillus fumigatus, seven Aspergillus lentulus, four Aspergillus wentii, two Aspergillus flavus, two Aspergillus terreus, one Aspergillus calidoustus, one Aspergillus westerdijkiae, one Aspergillus tamarii, and one Eurotium amstelodami. Aspergillus strains were evaluated for their susceptibility to medical azoles used in human therapy (itraconazole, posaconazole, and voriconazole) and to agricultural azoles (PEN) used in the prevention and treatment of plant diseases. The isolates showed moderate susceptibility to voriconazole. We did not observe any decrease of susceptibility to the medical azoles tested throughout the testing period in any of the treated plots, although some of the resistant species were isolated from there.