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Sample records for cell factory fishing

  1. 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.

  2. 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...

  3. 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...

  4. 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...

  5. 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....

  6. 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...

  7. 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...

  8. Fish germ cells

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Fish, like many other animals, have two major cell lineages, namely the germline and soma. The germ-soma separation is one of the earliest events of embryonic development. Germ cells can be specifically labeled and isolated for culture and transplan-tation, providing tools for reproduction of endangered species in close relatives, such as surrogate production of trout in salmon. Haploid cell cultures, such as medaka haploid embryonic stem cells have recently been obtained, which are capable of mimicking sperm to produce fertile offspring, upon nuclear being directly transferred into normal eggs. Such fish originated from a mosaic oocyte that had a haploid meiotic nucleus and a transplanted haploid mitotic cell culture nucleus. The first semi-cloned fish is Holly. Here we review the current status and future directions of understanding and manipulating fish germ cells in basic research and reproductive technology.

  9. 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....

  10. Fish stem cell cultures.

    Science.gov (United States)

    Hong, Ni; Li, Zhendong; Hong, Yunhan

    2011-04-13

    Stem cells have the potential for self-renewal and differentiation. First stem cell cultures were derived 30 years ago from early developing mouse embryos. These are pluripotent embryonic stem (ES) cells. Efforts towards ES cell derivation have been attempted in other mammalian and non-mammalian species. Work with stem cell culture in fish started 20 years ago. Laboratory fish species, in particular zebrafish and medaka, have been the focus of research towards stem cell cultures. Medaka is the second organism that generated ES cells and the first that gave rise to a spermatogonial stem cell line capable of test-tube sperm production. Most recently, the first haploid stem cells capable of producing whole animals have also been generated from medaka. ES-like cells have been reported also in zebrafish and several marine species. Attempts for germline transmission of ES cell cultures and gene targeting have been reported in zebrafish. Recent years have witnessed the progress in markers and procedures for ES cell characterization. These include the identification of fish homologs/paralogs of mammalian pluripotency genes and parameters for optimal chimera formation. In addition, fish germ cell cultures and transplantation have attracted considerable interest for germline transmission and surrogate production. Haploid ES cell nuclear transfer has proven in medaka the feasibility of semi-cloning as a novel assisted reproductive technology. In this special issue on "Fish Stem Cells and Nuclear Transfer", we will focus our review on medaka to illustrate the current status and perspective of fish stem cells in research and application. We will also mention semi-cloning as a new development to conventional nuclear transfer.

  11. Fish Stem Cell Cultures

    Directory of Open Access Journals (Sweden)

    Ni Hong, Zhendong Li, Yunhan Hong

    2011-01-01

    Full Text Available Stem cells have the potential for self-renewal and differentiation. First stem cell cultures were derived 30 years ago from early developing mouse embryos. These are pluripotent embryonic stem (ES cells. Efforts towards ES cell derivation have been attempted in other mammalian and non-mammalian species. Work with stem cell culture in fish started 20 years ago. Laboratory fish species, in particular zebrafish and medaka, have been the focus of research towards stem cell cultures. Medaka is the second organism that generated ES cells and the first that gave rise to a spermatogonial stem cell line capable of test-tube sperm production. Most recently, the first haploid stem cells capable of producing whole animals have also been generated from medaka. ES-like cells have been reported also in zebrafish and several marine species. Attempts for germline transmission of ES cell cultures and gene targeting have been reported in zebrafish. Recent years have witnessed the progress in markers and procedures for ES cell characterization. These include the identification of fish homologs/paralogs of mammalian pluripotency genes and parameters for optimal chimera formation. In addition, fish germ cell cultures and transplantation have attracted considerable interest for germline transmission and surrogate production. Haploid ES cell nuclear transfer has proven in medaka the feasibility of semi-cloning as a novel assisted reproductive technology. In this special issue on “Fish Stem Cells and Nuclear Transfer”, we will focus our review on medaka to illustrate the current status and perspective of fish stem cells in research and application. We will also mention semi-cloning as a new development to conventional nuclear transfer.

  12. Antibiotic resistance of Staphylococcus aureus strains isolated from fish processing factory workers

    OpenAIRE

    1988-01-01

    One hundred and twenty two strains of Staphylococcus aureus isolated from throats and palms of 39 workers from 6 fish processing factories situated in and around Cochin were tested for their sensitivity to nine commonly used antibiotics-ampicillin, chloramphenicol, erythromycin, kanamycin, neomycin, penicillin, polymyxin-B, streptomycin and tetracycline. Highest percentage of resistance was observed towards ampicillin followed by penicillin i.e. 64.75% and 59.84%. Resistance towards other ant...

  13. Fish as major carbonate mud producers and missing components of the tropical carbonate factory

    Science.gov (United States)

    Perry, Chris T.; Salter, Michael A.; Harborne, Alastair R.; Crowley, Stephen F.; Jelks, Howard L.; Wilson, Rod W.

    2011-01-01

    Carbonate mud is a major constituent of recent marine carbonate sediments and of ancient limestones, which contain unique records of changes in ocean chemistry and climate shifts in the geological past. However, the origin of carbonate mud is controversial and often problematic to resolve. Here we show that tropical marine fish produce and excrete various forms of precipitated (nonskeletal) calcium carbonate from their guts ("low" and "high" Mg-calcite and aragonite), but that very fine-grained (mostly 4 mole % MgCO3) are their dominant excretory product. Crystallites from fish are morphologically diverse and species-specific, but all are unique relative to previously known biogenic and abiotic sources of carbonate within open marine systems. Using site specific fish biomass and carbonate excretion rate data we estimate that fish produce ~6.1 x 106 kg CaCO3/year across the Bahamian archipelago, all as mud-grade (the factories function both today and in the past.

  14. 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.

  15. Fish as major carbonate mud producers and missing components of the tropical carbonate factory.

    Science.gov (United States)

    Perry, Chris T; Salter, Michael A; Harborne, Alastair R; Crowley, Stephen F; Jelks, Howard L; Wilson, Rod W

    2011-03-01

    Carbonate mud is a major constituent of recent marine carbonate sediments and of ancient limestones, which contain unique records of changes in ocean chemistry and climate shifts in the geological past. However, the origin of carbonate mud is controversial and often problematic to resolve. Here we show that tropical marine fish produce and excrete various forms of precipitated (nonskeletal) calcium carbonate from their guts ("low" and "high" Mg-calcite and aragonite), but that very fine-grained (mostly 4 mole % MgCO(3)) are their dominant excretory product. Crystallites from fish are morphologically diverse and species-specific, but all are unique relative to previously known biogenic and abiotic sources of carbonate within open marine systems. Using site specific fish biomass and carbonate excretion rate data we estimate that fish produce ∼6.1 × 10(6) kg CaCO(3)/year across the Bahamian archipelago, all as mud-grade (the fish to total carbonate mud production average ∼14% overall, and exceed 70% in specific habitats. Critically, we also document the widespread presence of these distinctive fish-derived carbonates in the finest sediment fractions from all habitat types in the Bahamas, demonstrating that these carbonates have direct relevance to contemporary carbonate sediment budgets. Fish thus represent a hitherto unrecognized but significant source of fine-grained carbonate sediment, the discovery of which has direct application to the conceptual ideas of how marine carbonate factories function both today and in the past.

  16. 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...

  17. Fish canning wastewater treatment by activated sludge: Application of factorial design optimization

    Directory of Open Access Journals (Sweden)

    Raquel O. Cristóvão

    2015-06-01

    Full Text Available The optimization of hydraulic retention time (HRT and initial organic matter concentration for dissolved organic carbon (DOC abatement of wastewater from a fish canning industry of northern Portugal by activated sludge was investigated using response surface methodology (RSM. The two parameters were chosen since it was found that the treatment efficiency is mainly influenced by them. The experimental data on DOC removal were fitted into a quadratic polynomial model using factorial design and RSM. The optimum process conditions were determined by analyzing the response surface of a three-dimensional plot and by solving the regression model equation. The obtained results showed a HRT of 6.4 h and an initial DOC of 406.2 mg/L as the best treatment conditions. Under these conditions, the maximum predicted DOC removal was 88.0%, confirming the feasibility and the reliability of fish canning wastewater treatment by activated sludge for organic content removal.

  18. 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...

  19. 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

  20. 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...

  1. 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

  2. From "Duck Factory" to "Fish Factory": Climate induced changes in vertebrate communities of prairie pothole wetlands and small lakes

    Science.gov (United States)

    McLean, Kyle I.; Mushet, David M.; Stockwell, Craig A.

    2016-01-01

    The Prairie Pothole Region’s myriad wetlands and small lakes contribute to its stature as the “duck factory” of North America. The fishless nature of the region’s aquatic habitats, a result of frequent drying, freezing, and high salinity, influences its importance to waterfowl. Recent precipitation increases have resulted in higher water levels and wetland/lake freshening. In 2012–13, we sampled chemical characteristics and vertebrates (fish and salamanders) of 162 Prairie Pothole wetlands and small lakes. We used non-metric multidimensional scaling, principal component analysis, and bootstrapping techniques to reveal relationships. We found fish present in a majority of sites (84 %). Fish responses to water chemistry varied by species. Fathead minnows (Pimephales promelas) and brook sticklebacks (Culaea inconstans) occurred across the broadest range of conditions. Yellow perch (Perca flavescens) occurred in a smaller, chemically defined, subset. Iowa darters (Etheostoma exile) were restricted to the narrowest range of conditions. Tiger salamanders (Ambystoma mavortium) rarely occurred in lakes with fish. We also compared our chemical data to similar data collected in 1966–1976 to explore factors contributing to the expansion of fish into previously fishless sites. Our work contributes to a better understanding of relationships between aquatic biota and climate-induced changes in this ecologically important area.

  3. 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...

  4. 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...

  5. The long shadow of our chemical past - High DDT concentrations in fish near a former agrochemicals factory in England.

    Science.gov (United States)

    Jürgens, Monika D; Crosse, John; Hamilton, Patrick B; Johnson, Andrew C; Jones, Kevin C

    2016-11-01

    A total of 81 roach (Rutilus rutilus) collected from 13 southern English river sites between 2007 and 2012, were analysed for organochlorine pesticides, PCBs, PBDEs and some metals. Unexpectedly high concentrations of the banned insecticide DDT and its degradation products DDE and DDD (∑DDTs) were found in the 10 fish from the river Lee (or Lea) which averaged 88 ± 70 (standard deviation) μg/kg ww, almost 20 times higher than the average for the remaining sites (4.8 ± 3.1 μg/kg). All fish from that site exceeded the Canadian Tissue Residue Guideline (environmental quality standard) of 14 μg/kg ∑DDTs. Concentrations of the insecticides chlordane and lindane as well as copper, which is often used as a fungicide, were also elevated in fish from the Lee, though not as much as those of DDTs. A likely explanation for these observations was found in a nearby former pesticide factory, which had stopped production about three decades earlier. An extensive review of recent literature data on DDT in wild European fish found that, while levels are now generally low, there were several other hotspots with ∑DDTs levels that may still be of concern.

  6. 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.

  7. 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....

  8. Influences of the environment on the endocrine and paracrine fish growth hormone-insulin-like growth factor-I system.

    Science.gov (United States)

    Reinecke, M

    2010-04-01

    Insulin-like growth factor-I (IGF-I) is a key component of the complex system that regulates differentiation, development, growth and reproduction of fishes. The IGF-I gene is mainly expressed in the liver that represents the principal source of endocrine IGF-I but also in numerous other organs where the hormone most probably acts in an autocrine-paracrine manner. The primary stimulus for synthesis and release of IGF-I is growth hormone (GH) from the anterior pituitary. Thus, in analogy to mammals, it is usual to speak of a fish 'GH-IGF-I axis'. The GH-IGF-I system is affected by changes in the environment and probably represents a target of endocrine disrupting compounds (EDC) that impair many physiological processes in fishes. Thus, the review deals with the influences of changes in different environmental factors, such as food availability, temperature, photoperiod, season, salinity and EDCs, on GH gene expression in pituitary, IGF-I gene expression in liver and extrahepatic sites and the physiological effects resulting from the evoked alterations in endocrine and local IGF-I. Environmental influences certainly interact with each other but for convenience of the reader they will be dealt with in separate sections. Current trends in GH-IGF-I research are analysed and future focuses are suggested at the end of the sections.

  9. [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.

  10. 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....

  11. 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.

  12. 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.

  13. 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

  14. 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...

  15. 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.

  16. 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...

  17. Fish T cells: recent advances through genomics

    Science.gov (United States)

    Laing, Kerry J.; Hansen, John D.

    2011-01-01

    This brief review is intended to provide a concise overview of the current literature concerning T cells, advances in identifying distinct T cell functional subsets, and in distinguishing effector cells from memory cells. We compare and contrast a wealth of recent progress made in T cell immunology of teleost, elasmobranch, and agnathan fish, to knowledge derived from mammalian T cell studies. From genome studies, fish clearly have most components associated with T cell function and we can speculate on the presence of putative T cell subsets, and the ability to detect their differentiation to form memory cells. Some recombinant proteins for T cell associated cytokines and antibodies for T cell surface receptors have been generated that will facilitate studying the functional roles of teleost T cells during immune responses. Although there is still a long way to go, major advances have occurred in recent years for investigating T cell responses, thus phenotypic and functional characterization is on the near horizon.

  18. 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.

  19. 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...

  20. 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...

  1. 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

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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....

  7. 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...

  8. 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...

  9. 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.

  10. 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....

  11. 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...

  12. 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.

  13. 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....

  14. Fish Stem Cells: Classification, Resources, Characteristics and Application Areas

    Directory of Open Access Journals (Sweden)

    Şehriban ÇEK

    2016-08-01

    Full Text Available Stem cells are a class of undifferentiated cells, have the potential for self-renewal that can differ to the specialized cells. First studies on stem cells in fish started with zebra fish in 1992. In this review, classification, resources, vital importance, characteristics and application areas of fish stem cell were clarified.

  15. Medaka fish stem cells and their applications

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Stem cells are present in developing embryos and adult tissues of multicellular organisms. Owing to their unique features, stem cells provide excellent opportunities for experimental analyses of basic developmental processes such as pluripotency control and cell fate decision and for regenerative medicine by stem cell-based therapy. Stem cell cultures have been best studied in 3 vertebrate organisms. These are the mouse, human and a small laboratory fish called medaka. Specifically, medaka has given rise to the first embryonic stem (ES) cells besides the mouse, the first adult testis-derived male stem cells spermatogonia capable of test-tube sperm production, and most recently, even haploid ES cells capable of producing Holly, a semi-cloned fertile female medaka from a mosaic oocyte created by microinjecting a haploid ES cell nucleus directly into a normal oocyte. These breakthroughs make medaka a favoring vertebrate model for stem cell research, the topic of this review.

  16. Sensitive-cell-based fish chromatophore biosensor

    Science.gov (United States)

    Plant, Thomas K.; Chaplen, Frank W.; Jovanovic, Goran; Kolodziej, Wojtek; Trempy, Janine E.; Willard, Corwin; Liburdy, James A.; Pence, Deborah V.; Paul, Brian K.

    2004-07-01

    A sensitive biosensor (cytosensor) has been developed based on color changes in the toxin-sensitive colored living cells of fish. These chromatophores are highly sensitive to the presence of many known and unknown toxins produced by microbial pathogens and undergo visible color changes in a dose-dependent manner. The chromatophores are immobilized and maintained in a viable state while potential pathogens multiply and fish cell-microbe interactions are monitored. Low power LED lighting is used to illuminate the chromatophores which are magnified using standard optical lenses and imaged onto a CCD array. Reaction to toxins is detected by observing changes is the total area of color in the cells. These fish chromatophores are quite sensitive to cholera toxin, Staphococcus alpha toxin, and Bordatella pertussis toxin. Numerous other toxic chemical and biological agents besides bacterial toxins also cause readily detectable color effects in chromatophores. The ability of the chromatophore cell-based biosensor to distinguish between different bacterial pathogens was examined. Toxin producing strains of Salmonella enteritis, Vibrio parahaemolyticus, and Bacillus cereus induced movement of pigmented organelles in the chromatophore cells and this movement was measured by changes in the optical density over time. Each bacterial pathogen elicited this measurable response in a distinctive and signature fashion. These results suggest a chromatophore cell-based biosensor assay may be applicable for the detection and identification of virulence activities associated with certain air-, food-, and water-borne bacterial pathogens.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. High average daily intake of PCDD/Fs and serum levels in residents living near a deserted factory producing pentachlorophenol (PCP) in Taiwan: influence of contaminated fish consumption

    Energy Technology Data Exchange (ETDEWEB)

    Lee Ching-Chang; Lin Wu-Ting; Liao Po-Chi; Su Huey-Jen [Dept. of Environmental and Occupational Health/Research Center of Environmental Trace Toxic substances, Medical Coll., National Cheng Kung Univ., Tainan (Taiwan); Chen Hsiu-Lin [Inst. of Basic Medical Sciences, Medical Coll., National Cheng Kung Univ., Tainan (Taiwan)

    2004-09-15

    Many reports have suggested that PCDD/Fs (polychlorinated dibenzo-p-dioxins and dibenzofurans) contribute to immune deficiency, liver damage, human carcinogenesis, and neuromotor maturation in children. Therefore, beginning in 1999, the Taiwan Environmental Protection Agency (EPA) conducted a survey to determine serum levels of PCDD/Fs in the general populations living around 19 incinerators in Taiwan. Relatively high average serum PCDD/F levels were unexpectedly found in Tainan city, a less industrialized area in southwestern Taiwan, than in other urban areas. We therefore reviewed the usage history of the land and found that a factory situated between Hsien-Gong Li and Lu-Erh Li, two administrative units of Tainan city, had been manufacturing pentachlorophenol (PCP) between 1967 and 1982. PCDD/Fs are formed as byproducts in the PCP manufacturing process. Exposure to PCP and its derivatives via the food chain is the most significant intake route of PCDD/Fs in consumers in the European Union (EU). In Japan, in addition to combustion processes, PCP and chlornitrofen (CNP) have also been identified as the major sources of PCDD/Fs in Tokyo Bay7. A preliminary investigation showed that the soil in the PCP factory and sediments in the sea reservoir (13 hectares) near the deserted factory were seriously contaminated with PCDD/Fs (260-184,000 and 20-6220 pg I-TEQ/g, respectively), levels higher than those in other countries. Therefore, the aim of this study was to compare the PCDD/F levels of fish meat in the sea reservoir and the serum in inhabitants living in the vicinity of the closed PCP plant and other nearby areas. The data from human and other biota samples might clarify the transmission pathway of the PCDD/F contaminants from the PCP factory to local residents, provide information about the exposure status of those living in the vicinity of the deserted PCP factory, and also lead to useful suggestions for controlling PCDD/F accumulation in those living near such

  2. 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...

  3. High average daily intake of PCDD/Fs and serum levels in residents living near a deserted factory producing pentachlorophenol (PCP) in Taiwan: Influence of contaminated fish consumption

    Energy Technology Data Exchange (ETDEWEB)

    Lee, C.C. [Department of Environmental and Occupational Health, Medical College, National Cheng Kung University, Tainan, Taiwan (China); Research Center of Environmental Trace Toxic Substances, Medical College, National Cheng Kung University, Tainan, Taiwan (China); Lin, W.T. [Department of Environmental and Occupational Health, Medical College, National Cheng Kung University, Tainan, Taiwan (China); Liao, P.C. [Department of Environmental and Occupational Health, Medical College, National Cheng Kung University, Tainan, Taiwan (China); Research Center of Environmental Trace Toxic Substances, Medical College, National Cheng Kung University, Tainan, Taiwan (China); Su, H.J. [Department of Environmental and Occupational Health, Medical College, National Cheng Kung University, Tainan, Taiwan (China); Research Center of Environmental Trace Toxic Substances, Medical College, National Cheng Kung University, Tainan, Taiwan (China); Chen, H.L. [Department of Industrial Safety and Health, Hung Kuang University, Taichung, 34 Chung Chie Rd. Sha Lu, Taichung 433, Taiwan (China)]. E-mail: hsiulin@sunrise.hk.edu.tw

    2006-05-15

    An abandoned pentachlorophenol plant and nearby area in southern Taiwan was heavily contaminated by dioxins, impurities formed in the PCP production process. The investigation showed that the average serum PCDD/Fs of residents living nearby area (62.5 pg WHO-TEQ/g lipid) was higher than those living in the non-polluted area (22.5 and 18.2 pg WHO-TEQ/g lipid) (P < 0.05). In biota samples, average PCDD/F of milkfish in sea reservoir (28.3 pg WHO-TEQ/g) was higher than those in the nearby fish farm (0.15 pg WHO-TEQ/g), and Tilapia and shrimp showed the similar trend. The average daily PCDD/Fs intake of 38% participants was higher than 4 pg WHO-TEQ/kg/day suggested by the world health organization. Serum PCDD/F was positively associated with average daily intake (ADI) after adjustment for age, sex, BMI, and smoking status. In addition, a prospective cohort study is suggested to determine the long-term health effects on the people living near factory. - Inhabitants living near a deserted PCP factory are exposed to high PCDD/F levels.

  4. 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...

  5. 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.

  6. 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.

  7. 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...

  8. [Zebra fish cell movements during gastrulation].

    Science.gov (United States)

    Zhang, Ting; Mo, Xian-Ming

    2013-04-01

    During zebrafish gastrulation, large cellular rearrangements create the formation of the three germ layers, ectoderm, mesoderm, and endoderm. This process includes three types of conserved morphogenetic movement: epiboly, involution, and convergent extension. Specially, the anterior movement of prechordal plate progenitors is essential for the location and differentiation of mesendoderm progenitors, and the pechordal plate progenitors'coherent migration is thought to be a good model to study the mechanism of cell movement in vivo. Gastrulation migration is known to be controlled by many signaling pathways such as Wnt/planar cell polarity signaling; however, the underlying molecular mechanism for cellular behavior remains unknown. At present, it is generally agree that cell adhesion and cytoskeletal rearrangement are critical factors during zebrafish gastrulation cell migration. In addition, the role of extraembryonic tissue (yolk syncytial layer) during gastrulation is concerned increasingly. Here, we described the essential factors for controlling cellular behaviors and highlighted the major issues and questions that require further investigation during zebra fish gastrular cell migration in order to provide a complete map containing all the factors for regulating gastrulation cell migration and their interactions on a cellular level.

  9. 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.

  10. 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

  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. 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.

  15. 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.

  16. 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.

  17. 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

  18. 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.

  19. 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.

  20. 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...

  1. 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$\

  2. Regulation of stem cells in the zebra fish hematopoietic system.

    Science.gov (United States)

    Huang, H-T; Zon, L I

    2008-01-01

    Hematopoietic stem cells (HSCs) have been used extensively as a model for stem cell biology. Stem cells share the ability to self-renew and differentiate into multiple cell types, making them ideal candidates for tissue regeneration or replacement therapies. Current applications of stem cell technology are limited by our knowledge of the molecular mechanisms that control their proliferation and differentiation, and various model organisms have been used to fill these gaps. This chapter focuses on the contributions of the zebra fish model to our understanding of stem cell regulation within the hematopoietic system. Studies in zebra fish have been valuable for identifying new genetic and signaling factors that affect HSC formation and development with important implications for humans, and new advances in the zebra fish toolbox will allow other aspects of HSC behavior to be investigated as well, including migration, homing, and engraftment.

  3. Relationship Between Hair Cell Loss and Hearing Loss in Fishes.

    Science.gov (United States)

    Smith, Michael E

    2016-01-01

    Exposure to intense sound or ototoxic chemicals can damage the auditory hair cells of vertebrates, resulting in hearing loss. Although the relationship between such hair cell damage and auditory function is fairly established for terrestrial vertebrates, there are limited data available to understand this relationship in fishes. Although investigators have measured either the morphological damage of the inner ear or the functional deficits in the hearing of fishes, very few have directly measured both in an attempt to find a relationship between the two. Those studies that have examined both auditory hair cell damage in the inner ear and the resulting hearing loss in fishes are reviewed here. In general, there is a significant linear relationship between the number of hair cells lost and the severity of hearing threshold shifts, although this varies between species and different hair cell-damaging stimuli. After trauma to the fish ear, auditory hair cells are able to regenerate to control level densities. With this regeneration also comes a restoration of hearing. Thus there is also a significant relationship between hair cell recovery and hearing recovery in fishes.

  4. 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.

  5. 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.

  6. Sensory hair cell death and regeneration in fishes

    Directory of Open Access Journals (Sweden)

    Jerry D. Monroe

    2015-04-01

    Full Text Available Sensory hair cells are specialized mechanotransductive receptors required for hearing and vestibular function. Loss of hair cells in humans and other mammals is permanent and causes reduced hearing and balance. In the early 1980’s, it was shown that hair cells continue to be added to the inner ear sensory epithelia in cartilaginous and bony fishes. Soon thereafter, hair cell regeneration was documented in the chick cochlea following acoustic trauma. Since then, research using chick and other avian models has led to great insights into hair cell death and regeneration. However, with the rise of the zebrafish as a model organism for studying disease and developmental processes, there has been an increased interest in studying sensory hair cell death and regeneration in its lateral line and inner ears. Advances derived from studies in zebrafish and other fish species include understanding the effect of ototoxins on hair cells and finding otoprotectants to mitigate ototoxin damage, the role of cellular proliferation versus direct transdifferentiation during hair cell regeneration, and elucidating cellular pathways involved in the regeneration process. This review will summarize research on hair cell death and regeneration using fish models, indicate the potential strengths and weaknesses of these models, and discuss several emerging areas of future studies.

  7. 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.

  8. Enteroendocrine cells of the cyprinid fish, Barbus conchonius

    NARCIS (Netherlands)

    Rombout, J.H.W.M.

    1980-01-01

    Information on the endocrine regulation of digestion in fish is scarce especially on stomachless cyprinids. In the present study (chapters I, III) 3 distinct enteroendocrine cell types will be described for the intestinal epithelium of Barbus conchonius. With the light microscope, enteroendocrine ce

  9. 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.

  10. 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.

  11. 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.

  12. 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...

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. Fish peripheral blood mononuclear cells preparation for future monitoring applications.

    Science.gov (United States)

    Pierrard, Marie-Aline; Roland, Kathleen; Kestemont, Patrick; Dieu, Marc; Raes, Martine; Silvestre, Frédéric

    2012-07-15

    Fish species possess many specific characteristics that support their use in ecotoxicology. Widely used in clinical research, peripheral blood mononuclear cells (PBMCs) can reasonably be exploited as relevant target cells in the assessment of environmental chemical toxicity. The current article focuses on the methods necessary to isolate, characterize, and culture fish PBMCs. These procedures were successfully applied on an endangered species, the European eel (Anguilla anguilla L.), and on an economically important and worldwide exported species, the Asian catfish (Pangasianodon hypophthalmus S.). Proteomic approaches can be useful to screen xenobiotic exposure at the protein expression level, giving the opportunity to develop early warning signals thanks to molecular signatures of toxicity. To date, a major limitation of proteomic analyses is that most protein expression profiles often reveal the same predominant and frequently differentially expressed families of proteins regardless of the experimental stressing conditions. The current study describes a methodology to get a postnuclear fraction of high quality isolated from fish PBMCs in order to perform subsequent subproteomic analyses. Applied on samples from eel, the subproteomic analysis (two-dimensional differential in-gel electrophoresis) allowed the identification by liquid chromatography-tandem mass spectrometry and searches in the full NCBInr (National Center for Biotechnology Information nonredundant) database of 66 proteins representing 36 different proteins validated through Peptide and Protein Prophet of Scaffold software.

  19. 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.

  20. Establishment and characterization of a new marine fish cell line from ovary of barfin flounder ( Verasper moseri)

    Science.gov (United States)

    Xu, Xiaohui; Fan, Tingjun; Jiang, Guojian; Yang, Xiuxia

    2015-12-01

    A novel continuous ovary cell line from barfin flounder ( Verasper moseri) (BFO cell line) was established with its primitive application in transgenic expression demonstrated in this study. Primarily cultured cells grew well at 22°C in Dulbecco's modified Eagle medium/F12 medium (DMEM/F12, 1:1; pH 7.2) supplemented with 20% fetal bovine serum (FBS), carboxymethyl chitooligosaccharide, basic fibroblast growth factor (bFGF) and insulin-like growth factor-I (IGF-I). The primary BFO cells in fibroblastic morphology proliferated into a confluent monolayer about 2 weeks later, and were able to be subcultured. Impacts of medium and temperature on the growth of the cells were examined. The optimum growth was found in DMEM/F12 with 20% FBS and at 22°C. The BFO cells can be continuously subcultured to Passage 120 steadily with a population doubling time of 32.7 h at Passage 60. Chromosome analysis revealed that 72% of BFO cells at Passage 60 maintained the normal diploid chromosome number (46) with a normal karyotype of 2st+44t. The results of gene transformation indicated that green fluorescence protein (GFP) positively expressed in these cells after being transformed with pcDNA3.1-GFP. Therefore, a continuous and transformable BFO cell line was successfully established, which may serve as a useful tool for cytotechnological manipulation and transgenic modification of this fish.

  1. Fishing

    Institute of Scientific and Technical Information of China (English)

    姜群山

    2002-01-01

    @@ Last Saturday my cousin (表兄) came to my home. We were very happy to see each other. We decided that the next day we went to fish. We got up very early that day. When we left home,the moon could still be seen in the sky.

  2. 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.

  3. Studies on the Developmental Potentiality of Cultured Cell Nuclei of Fish

    Directory of Open Access Journals (Sweden)

    Hongxi Chen, Yonglan Yi, Minrong Chen, Xingqi Yang

    2010-01-01

    Full Text Available By means of the serial nuclear transplantation technique, the authors obtained a nuclear transplant fish from subcultured cell originated from the blastula cells of the crucian carp (Carassius auratus Linnaeus. This nuclear transplant fish survived for three years, but its sexual glands were undifferentiated. The authors have also obtained a sexually mature adult fish from short-term cultured kidney cell nucleus of an adult crucian carp. Results of the experiment implied that the subcultured cell nuclei of fish blastula cells and the specialized somatic cell nuclei of adult fish still retained their developmental totipotency, and thus, it indicated that there is a possibility of fish somatic cell breeding through the use of nuclear transplantation.

  4. 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.

  5. 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.

  6. 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....

  7. Germ cell transplantation as a potential biotechnological approach to fish reproduction.

    Science.gov (United States)

    Lacerda, S M S N; Costa, G M J; Campos-Junior, P H A; Segatelli, T M; Yazawa, R; Takeuchi, Y; Morita, T; Yoshizaki, G; França, L R

    2013-02-01

    Although the use of germ cell transplantation has been relatively well established in mammals, the technique has only been adapted for use in fish after entering the 2000s. During the last decade, several different approaches have been developed for germ cell transplantation in fish using recipients of various ages and life stages, such as blastula-stage embryos, newly hatched larvae and sexually mature specimens. As germ cells can develop into live organisms through maturation and fertilization processes, germ cell transplantation in fish has opened up new avenues of research in reproductive biotechnology and aquaculture. For instance, the use of xenotransplantation in fish has lead to advances in the conservation of endangered species and the production of commercially valuable fish using surrogated recipients. Further, this could also facilitate the engineering of transgenic fish. However, as is the case with mammals, knowledge regarding the basic biology and physiology of germline stem cells in fish remains incomplete, imposing a considerable limitation on the application of germ cell transplantation in fish. Furthering our understanding of germline stem cells would contribute significantly to advances regarding germ cell transplantation in fish.

  8. Silver nanoparticles disrupt regulation of steroidogenesis in fish ovarian cells.

    Science.gov (United States)

    Degger, Natalie; Tse, Anna C K; Wu, Rudolf S S

    2015-12-01

    Despite the influx of silver nanoparticles (nAg) into the marine environment, their effects on fish reproduction remain completely unexplored. Using ovarian primary cells from marine medaka (Oryzias melastigma), in vitro studies were carried out to evaluate the effects of two differently coated nAg particles (Oleic Acid, (OA) nAg and Polyvinylpyrrolidone, (PVP) nAg) on fish ovarian tissues, using AgNO3 as a positive control. Cytotoxicity was evaluated by MTT assay and expression of key genes regulating steroidogenesis (StAR, CYP 19a, CYP 11a, 3βHSD and 20βHSD) were determined by Q-RT-PCR. EC50 values for PVP nAg, OA nAg and AgNO3 were 7.25μgL(-1), 924.4μgL(-1), and 42.0μgL(-1) respectively, showing that toxicity of silver was greatly enhanced in the PVP coated nano-form. Down regulation of CYP 19a was observed in both nAg and AgNO3 treatments, while down regulation of 3βHSD was only found in the OA nAg and AgNO3 treatments. For the first time, our results demonstrated that nAg can affect specific genes regulating steroidogenesis, implicating nAg as a potential endocrine disruptor.

  9. 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

  10. 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...

  11. DNA breakage detection-fluorescence in situ hybridization (DBD-FISH in buccal cells

    Directory of Open Access Journals (Sweden)

    E. I. Cortés-Gutiérrez

    2012-12-01

    Full Text Available DNA breakage detection-fluorescence in situ hybridization (DBD-FISH is a recently developed technique that allows cell-by-cell detection and quantification of DNA breakage in the whole genome or within specific DNA sequences. The present investigation was conducted to adapt the methodology of DBD-FISH to the visualization and evaluation of DNA damage in buccal epithelial cells. DBD-FISH revealed that DNA damage increased significantly according to H2O2 concentration (r2=0.91. In conclusion, the DBD-FISH technique is easy to apply in buccal cells and provides prompt results that are easy to interpret. Future studies are needed to investigate the potential applicability of a buccal cell DBD-FISH model to human biomonitoring and nutritional work.

  12. DNA breakage detection-fluorescence in situ hybridization (DBD-FISH) in buccal cells.

    Science.gov (United States)

    Cortés-Gutiérrez, E I; Dávila-Rodríguez, M I; Fernández, J L; López-Fernández, C; Gosálvez, J

    2012-12-28

    DNA breakage detection-fluorescence in situ hybridization (DBD-FISH) is a recently developed technique that allows cell-by-cell detection and quantification of DNA breakage in the whole genome or within specific DNA sequences. The present investigation was conducted to adapt the methodology of DBD-FISH to the visualization and evaluation of DNA damage in buccal epithelial cells. DBD-FISH revealed that DNA damage increased significantly according to H2O2 concentration (r2=0.91). In conclusion, the DBD-FISH technique is easy to apply in buccal cells and provides prompt results that are easy to interpret. Future studies are needed to investigate the potential applicability of a buccal cell DBD-FISH model to human biomonitoring and nutritional work.

  13. 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...

  14. Shedding light on proteins, nucleic acids, cells, humans and fish

    Science.gov (United States)

    Setlow, Richard B.

    2002-01-01

    I was trained as a physicist in graduate school. Hence, when I decided to go into the field of biophysics, it was natural that I concentrated on the effects of light on relatively simple biological systems, such as proteins. The wavelengths absorbed by the amino acid subunits of proteins are in the ultraviolet (UV). The wavelengths that affect the biological activities, the action spectra, also are in the UV, but are not necessarily parallel to the absorption spectra. Understanding these differences led me to investigate the action spectra for affecting nucleic acids, and the effects of UV on viruses and cells. The latter studies led me to the discovery of the important molecular nature of the damages affecting DNA (cyclobutane pyrimidine dimers) and to the discovery of nucleotide excision repair. Individuals with the genetic disease xeroderma pigmentosum (XP) are extraordinarily sensitive to sunlight-induced skin cancer. The finding, by James Cleaver, that their skin cells were defective in DNA repair strongly suggested that DNA damage was a key step in carcinogenesis. Such information was important for estimating the wavelengths in sunlight responsible for human skin cancer and for predicting the effects of ozone depletion on the incidence of non-melanoma skin cancer. It took experiments with backcross hybrid fish to call attention to the probable role of the longer UV wavelengths not absorbed by DNA in the induction of melanoma. These reflections trace the biophysicist's path from molecules to melanoma.

  15. Production of reproductively sterile fish: A mini-review of germ cell elimination technologies.

    Science.gov (United States)

    Wong, Ten-Tsao; Zohar, Yonathan

    2015-09-15

    As seafood consumption shifts from fisheries harvests to artificially propagated aquatic species, the increase of aquaculture activities poses a biological threat to our environment. Selectively bred, non-native and (eventually) genetically engineered farmed fish may escape from aquaculture operations, propagate and/or interbreed with wild stocks and subsequently alter the genetic makeup of populations in the environment. Thus, an effective strategy for bio-containment of farmed fish is critically needed. Farming reproductively sterile fish is the most environmentally sustainable approach to ensure complete bio-containment in large-scale aquaculture operations. Chromosome set manipulations to produce sterile fish, including polyploidy and hybridization, are currently the most common practices in the aquaculture industry. However, they do not always result in 100% sterility of the treated fish. Moreover, triploid fish typically do not perform as well as the non-manipulated diploids under commercial culture conditions. In the last half decade, several genetic engineering methods have been developed to produce sterile fish. In this review, we will address the latest technologies that use transgenic approaches to eliminate germ cells, resulting in the production of sterile fish. These latest advances also led us to the development of egg/embryo immersion methodologies to deliver and screen compounds that can be used to eliminate primordial germ cells and produce sterile fish. This emerging non-transgenic strategy for the production of reproductively sterile fish in aquaculture will also be discussed.

  16. The astonishing diversity of Ig classes and B cell repertoires in teleost fish

    Directory of Open Access Journals (Sweden)

    Simon eFillatreau

    2013-02-01

    Full Text Available With lympoid tissue anatomy different than mammals, and diverse adaptations to all aquatic environments, fish constitute a fascinating group of vertebrate to study the biology of B cell repertoires in a comparative perspective. Fish B lymphocytes express immunoglobulin (Ig on their surface and secrete antigen-specific antibodies in response to immune challenges. Three antibody classes have been identified in fish, namely IgM, IgD and IgT, while IgG, IgA and IgE are absent. IgM and IgD have been found in all fish species analyzed, and thus seem to be primordial antibody classes. IgM and IgD are normally co-expressed from the same mRNA through alternative splicing, as in mammals. Tetrameric IgM is the main antibody class found in serum. Some species of fish also have IgT, which seems to exist only in fish and is specialized in mucosal immunity. IgM/IgD and IgT are expressed by two different sub-populations of B cells. The tools available to investigate B cell responses at the cellular level in fish are limited, but the progress of fish genomics has started to unravel a rich diversity of IgH and IgL locus organization, which might be related to the succcession of genome remodellings that occured during fish evolution. Moreover, the development of deep sequencing techniques has allowed the investigation of the global features of the expressed fish B cell repertoires in zebrafish and rainbow trout, in steady state or after infection. This review provides a description of the organization of fish Ig loci, with a particular emphasis on their heterogeneity between species, and presents recent data on the structure of the expressed Ig repertoire in healthy and infected fish.

  17. Mos1 transposon-based transformation of fish cell lines using baculoviral vectors

    Energy Technology Data Exchange (ETDEWEB)

    Yokoo, Masako [Laboratory of Applied Molecular Entomology, Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589 (Japan); Fujita, Ryosuke [Laboratory of Applied Molecular Entomology, Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589 (Japan); Innate Immunity Laboratory, Graduate School of Life Science and Creative Research Institution, Hokkaido University, Sapporo 001-0021 (Japan); Nakajima, Yumiko [Functional Genomics Group, COMB, Tropical Biosphere Research Center, University of the Ryukyus, Okinawa 903-0213 (Japan); Yoshimizu, Mamoru; Kasai, Hisae [Faculty of Fisheries Sciences, Hokkaido University, Hakodate 041-8611 (Japan); Asano, Shin-ichiro [Laboratory of Applied Molecular Entomology, Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589 (Japan); Bando, Hisanori, E-mail: hban@abs.agr.hokudai.ac.jp [Laboratory of Applied Molecular Entomology, Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589 (Japan)

    2013-09-13

    Highlights: •The baculovirus vector infiltrates the cells of economic important fishes. •Drosophila Mos1 transposase expressed in fish cells maintains its ability to localize to the nucleus. •The baculoviral vector carrying Mos1 is a useful tool to stably transform fish cells. -- Abstract: Drosophila Mos1 belongs to the mariner family of transposons, which are one of the most ubiquitous transposons among eukaryotes. We first determined nuclear transportation of the Drosophila Mos1-EGFP fusion protein in fish cell lines because it is required for a function of transposons. We next constructed recombinant baculoviral vectors harboring the Drosophila Mos1 transposon or marker genes located between Mos1 inverted repeats. The infectivity of the recombinant virus to fish cells was assessed by monitoring the expression of a fluorescent protein encoded in the viral genome. We detected transgene expression in CHSE-214, HINAE, and EPC cells, but not in GF or RTG-2 cells. In the co-infection assay of the Mos1-expressing virus and reporter gene-expressing virus, we successfully transformed CHSE-214 and HINAE cells. These results suggest that the combination of a baculovirus and Mos1 transposable element may be a tool for transgenesis in fish cells.

  18. 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

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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...

  4. 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...

  5. FISH studies of chromosome abnormalities in germ cells and its relevance in reproductive counseling

    Institute of Scientific and Technical Information of China (English)

    Zaida Sarrate; Joan Blanco; Ester Anton; Susana Egozcue; Josep Egozcue; Francesca Vidal

    2005-01-01

    Chromosome abnormalities are one of the major causes of human infertility. In infertile males, abnormal karyotypes are more frequent than in the general population. Furthermore, meiotic disorders affecting the germ cell-line have been observed in men with normal somatic karyotypes consulting for infertility. In both cases, the production of unbalanced spermatozoa has been demonstrated. Basically addressed to establish reproductive risks, fluorescence in situ hybridization (FISH) on decondensed sperm heads has become the most frequently used method to evaluate the chromosomal constitution of spermatozoa in carriers of numerical sex chromosome abnormalities, carriers of structural chromosome reorganizations and infertile males with normal karyotype. The aim of this review is to present updated figures of the information obtained through sperm FISH studies with an emphasis on its clinical significance. Furthermore, the incorporation of novel FISH-based techniques (Multiplex-FISH; Multi-FISH) in male infertility studies is also discussed.

  6. 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.

  7. 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.

  8. Single-cell transcriptome analysis of fish immune cells provides insight into the evolution of vertebrate immune cell types

    Science.gov (United States)

    Ferreira, Lauren; Macaulay, Iain C.; Stubbington, Michael J.T.

    2017-01-01

    The immune system of vertebrate species consists of many different cell types that have distinct functional roles and are subject to different evolutionary pressures. Here, we first analyzed conservation of genes specific for all major immune cell types in human and mouse. Our results revealed higher gene turnover and faster evolution of trans-membrane proteins in NK cells compared with other immune cell types, and especially T cells, but similar conservation of nuclear and cytoplasmic protein coding genes. To validate these findings in a distant vertebrate species, we used single-cell RNA sequencing of lck:GFP cells in zebrafish and obtained the first transcriptome of specific immune cell types in a nonmammalian species. Unsupervised clustering and single-cell TCR locus reconstruction identified three cell populations, T cells, a novel type of NK-like cells, and a smaller population of myeloid-like cells. Differential expression analysis uncovered new immune-cell–specific genes, including novel immunoglobulin-like receptors, and neofunctionalization of recently duplicated paralogs. Evolutionary analyses confirmed the higher gene turnover of trans-membrane proteins in NK cells compared with T cells in fish species, suggesting that this is a general property of immune cell types across all vertebrates. PMID:28087841

  9. FISH 'N' Chips : a single cell genomic analyzer for the human microbiome.

    Energy Technology Data Exchange (ETDEWEB)

    Light, Yooli Kim; Perroud, Thomas D.; Hugenholtz, Philip (Joint Genome Institute, Walnut Creek, CA); Meagher, Robert J.; Singh, Anup K.; Malamud, Daniel (New York University, New York, NY); Saxena, Deepak (New York University, New York, NY); Liu, Peng

    2010-09-01

    Uncultivable microorganisms likely play significant roles in the ecology within the human body, with subtle but important implications for human health. Focusing on the oral microbiome, we are developing a processor for targeted isolation of individual microbial cells, facilitating whole-genome analysis without the need for isolation of pure cultures. The processor consists of three microfluidic modules: identification based on 16S rRNA fluorescence in situ hybridization (FISH), fluorescence-based sorting, and encapsulation of individual selected cells into small droplets for whole genome amplification. We present here a technique for performing microscale FISH and flow cytometry, as a prelude to single cell sorting.

  10. Causes and Consequences of Sensory Hair Cell Damage and Recovery in Fishes.

    Science.gov (United States)

    Smith, Michael E; Monroe, J David

    2016-01-01

    Sensory hair cells are the mechanotransductive receptors that detect gravity, sound, and vibration in all vertebrates. Damage to these sensitive receptors often results in deficits in vestibular function and hearing. There are currently two main reasons for studying the process of hair cell loss in fishes. First, fishes, like other non-mammalian vertebrates, have the ability to regenerate hair cells that have been damaged or lost via exposure to ototoxic chemicals or acoustic overstimulation. Thus, they are used as a biomedical model to understand the process of hair cell death and regeneration and find therapeutics that treat or prevent human hearing loss. Secondly, scientists and governmental natural resource managers are concerned about the potential effects of intense anthropogenic sounds on aquatic organisms, including fishes. Dr. Arthur N. Popper and his students, postdocs and research associates have performed pioneering experiments in both of these lines of fish hearing research. This review will discuss the current knowledge regarding the causes and consequences of both lateral line and inner ear hair cell damage in teleost fishes.

  11. "Fish-in-net", a novel method for cell immobilization of Zymomonas mobilis.

    Directory of Open Access Journals (Sweden)

    Xuedun Niu

    Full Text Available BACKGROUND: Inorganic mesoporous materials exhibit good biocompatibility and hydrothermal stability for cell immobilization. However, it is difficult to encapsulate living cells under mild conditions, and new strategies for cell immobilization are needed. We designed a "fish-in-net" approach for encapsulation of enzymes in ordered mesoporous silica under mild conditions. The main objective of this study is to demonstrate the potential of this approach in immobilization of living cells. METHODOLOGY/PRINCIPAL FINDINGS: Zymomonas mobilis cells were encapsulated in mesoporous silica-based materials under mild conditions by using a "fish-in-net" approach. During the encapsulation process, polyethyleneglycol was used as an additive to improve the immobilization efficiency. After encapsulation, the pore size, morphology and other features were characterized by various methods, including scanning electron microscopy, nitrogen adsorption-desorption analysis, transmission electron microscopy, fourier transform infrared spectroscopy, and elemental analysis. Furthermore, the capacity of ethanol production by immobilized Zymomonas mobilis and free Zymomonas mobilis was compared. CONCLUSIONS/SIGNIFICANCE: In this study, Zymomonas mobilis cells were successfully encapsulated in mesoporous silica-based materials under mild conditions by the "fish-in-net" approach. Encapsulated cells could perform normal metabolism and exhibited excellent reusability. The results presented here illustrate the enormous potential of the "fish-in-net" approach for immobilization of living cells.

  12. Voltage-gated sodium channels in cerebellar Purkinje cells of mormyrid fish

    NARCIS (Netherlands)

    M.M. de Ruiter (Martijn); C.I. de Zeeuw (Chris); C.R.W. Hansel (Christian)

    2006-01-01

    textabstractCerebellar Purkinje cells of mormyrid fish differ in some morphological as well as physiological parameters from their counterparts in mammals. Morphologically, Purkinje cells of mormyrids have larger dendrites that are characterized by a lower degree of branching in the molecular layer.

  13. 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.

  14. 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...

  15. Effect of the pesticide Deltamethrin on the Mauthner cells of Lake Balaton fish.

    Science.gov (United States)

    Csillik, B; Fazakas, J; Nemcsók, J; Knyihár-Csillik, E

    2000-06-01

    Deltamethrin, a synthetic pesticide [(S)alpha-cyano-3-phenoxybenzyl-(1R)-cis-3-(2.2-dibromovinyl)-2,2-dim ethylcyclopropane-carboxylate] used for extermination of mosquitoes on the shores of lake Balaton, has been found to induce severe impairments of the nervous system of several Lake Balaton fish, such as carp (Cyprinus carpio), goldfish (Carassius auratus gibelis Bloch), eel (Anguilla anguilla) and wels (Silurus glanis). It has been shown that Deltamethrin, in a concentration of 1 microgram/liter in the aquarium water, inhibits acetylcholinesterase enzyme activity of the giant Mauthner's nerve cells as well as of the axon terminals synapsing with these cells. Even more importantly, however, Deltamethrin in a concentration of 10 micrograms per liter, induces blockade of the expression of choline acetyltransferase in the bulbous axon terminals synapsing with the lateral dendrites of the Mauthner cells. Since, under normal conditions, the function of the Mauthner cells is to co-ordinate the C-start reaction, by which fish rapidly leave sites of nociceptive stimulation, it stands for reason to assume that Deltamethrin intoxicated fish may be prone to become victims of various factors which endanger survival of the individual. During the last decade, waves of fish deaths were observed in Lake Balaton, which is the largest fresh-water lake in Europe. Fish death coincided with airborne mosquito-killing campaigns. Results of the enzyme- and immunohistochemical studies described in this paper, together with the deleterious effects of Deltamethrin to the enteric nervous system of fish which has been reported earlier (Lang et al., 1997) suggest that fish death might be caused by the indiscriminate use of Deltamethrin airborne spray in the mosquito-extermination campaigns.

  16. APPLICATION OF TWO-COLOR INTERPHASE FISH USING SEX PROBE IN ALLOGENEIC STEM CELL TRANSPLANTATION

    Institute of Scientific and Technical Information of China (English)

    曾慧兰; 李建勇; 朱康儿; 薛永权; 李杨秋; 刘晓力; 过宇

    2002-01-01

    Objective: To evaluate the significance of two-color interphase fluorescence in situ hybridization (FISH) using X and Y centromere probe in the engraftment estimation and minimal residual disease (MRD) monitoring after allogeneic stem cell transplantation (alloSCT). Methods: Samples from 12 cases patients in different periods after alloSCT were detected by interphase FISH. Results: All of the 12 patients were proved to obtain engraftment 22(35 days after alloSCT. While traditional karyotype showed as 100%XX or 100%XY invariably, FISH showed different percentages of donor original sex chromosome. Conclusion: Two-color interphase FISH is a more sensitive and simple test for engraftment evaluation and MRD monitoring post SCT, though, it can not entirely replace traditional karyotype analysis and gene detection by RT-PCR.

  17. Identification and Characterization of MicroRNAs in Snakehead Fish Cell Line upon Snakehead Fish Vesiculovirus Infection.

    Science.gov (United States)

    Liu, Xiaodan; Tu, Jiagang; Yuan, Junfa; Liu, Xueqin; Zhao, Lijuan; Dawar, Farman Ullah; Khattak, Muhammad Nasir Khan; Hegazy, Abeer M; Chen, Nan; Vakharia, Vikram N; Lin, Li

    2016-01-26

    MicroRNAs (miRNAs) play important roles in mediating multiple biological processes in eukaryotes and are being increasingly studied to evaluate their roles associated with cellular changes following viral infection. Snakehead fish Vesiculovirus (SHVV) has caused mass mortality in snakehead fish during the past few years. To identify specific miRNAs involved in SHVV infection, we performed microRNA deep sequencing on a snakehead fish cell line (SSN-1) with or without SHVV infection. A total of 205 known miRNAs were identified when they were aligned with the known zebrafish miRNAs, and nine novel miRNAs were identified using MiRDeep2 software. Eighteen and 143 of the 205 known miRNAs were differentially expressed at three and 24 h post-infection (poi), respectively. From the differentially-expressed miRNAs, five were randomly selected to validate their expression profiles using quantitative reverse transcription polymerase chain reaction (qRT-PCR), and their expression profiles were consistent with the microRNA sequencing results. In addition, the target gene prediction of the SHVV genome was performed for the differentially-expressed host miRNAs, and a total of 10 and 58 differentially-expressed miRNAs were predicted to bind to the SHVV genome at three and 24 h poi, respectively. The effects of three selected miRNAs (miR-130-5p, miR-214 and miR-216b) on SHVV multiplication were evaluated using their mimics and inhibitors via qRT-PCR and Western blotting. The results showed that all three miRNAs were able to inhibit the multiplication of SHVV; whereas the mechanisms underlying the SHVV multiplication inhibited by the specific miRNAs need to be further characterized in the future.

  18. Identification and Characterization of MicroRNAs in Snakehead Fish Cell Line upon Snakehead Fish Vesiculovirus Infection

    Directory of Open Access Journals (Sweden)

    Xiaodan Liu

    2016-01-01

    Full Text Available MicroRNAs (miRNAs play important roles in mediating multiple biological processes in eukaryotes and are being increasingly studied to evaluate their roles associated with cellular changes following viral infection. Snakehead fish Vesiculovirus (SHVV has caused mass mortality in snakehead fish during the past few years. To identify specific miRNAs involved in SHVV infection, we performed microRNA deep sequencing on a snakehead fish cell line (SSN-1 with or without SHVV infection. A total of 205 known miRNAs were identified when they were aligned with the known zebrafish miRNAs, and nine novel miRNAs were identified using MiRDeep2 software. Eighteen and 143 of the 205 known miRNAs were differentially expressed at three and 24 h post-infection (poi, respectively. From the differentially-expressed miRNAs, five were randomly selected to validate their expression profiles using quantitative reverse transcription polymerase chain reaction (qRT-PCR, and their expression profiles were consistent with the microRNA sequencing results. In addition, the target gene prediction of the SHVV genome was performed for the differentially-expressed host miRNAs, and a total of 10 and 58 differentially-expressed miRNAs were predicted to bind to the SHVV genome at three and 24 h poi, respectively. The effects of three selected miRNAs (miR-130-5p, miR-214 and miR-216b on SHVV multiplication were evaluated using their mimics and inhibitors via qRT-PCR and Western blotting. The results showed that all three miRNAs were able to inhibit the multiplication of SHVV; whereas the mechanisms underlying the SHVV multiplication inhibited by the specific miRNAs need to be further characterized in the future.

  19. Susceptibility testing of fish cell lines for virus isolation

    DEFF Research Database (Denmark)

    Ariel, Ellen; Skall, Helle Frank; Olesen, Niels Jørgen

    2009-01-01

    compare susceptibility between cell lines and between lineages within a laboratory and between laboratories (Inter-laboratory Proficiency Test). The objective being that the most sensitive cell line and lineages are routinely selected for diagnostic purposes.In comparing cell lines, we simulated "non......-cell-culture-adapted" virus by propagating the virus in heterologous cell lines to the one tested. A stock of test virus was produced and stored at - 80 °C and tests were conducted biannually. This procedure becomes complicated when several cell lines are in use and does not account for variation among lineages. In comparing...... cell lineages, we increased the number of isolates of each virus, propagated stocks in a given cell line and tested all lineages of that line in use in the laboratory. Testing of relative cell line susceptibility between laboratories is carried out annually via the Inter-laboratory Proficiency Test...

  20. Genetic resistance to rhabdovirus infection in teleost fish is paralleled to the derived cell resistance status.

    Directory of Open Access Journals (Sweden)

    Eloi R Verrier

    Full Text Available Genetic factors of resistance and predisposition to viral diseases explain a significant part of the clinical variability observed within host populations. Predisposition to viral diseases has been associated to MHC haplotypes and T cell immunity, but a growing repertoire of innate/intrinsic factors are implicated in the genetic determinism of the host susceptibility to viruses. In a long-term study of the genetics of host resistance to fish rhabdoviruses, we produced a collection of double-haploid rainbow trout clones showing a wide range of susceptibility to Viral Hemorrhagic Septicemia Virus (VHSV waterborne infection. The susceptibility of fibroblastic cell lines derived from these clonal fish was fully consistent with the susceptibility of the parental fish clones. The mechanisms determining the host resistance therefore did not associate with specific host immunity, but rather with innate or intrinsic factors. One cell line was resistant to rhabdovirus infection due to the combination of an early interferon IFN induction--that was not observed in the susceptible cells--and of yet unknown factors that hamper the first steps of the viral cycle. The implication of IFN was well consistent with the wide range of resistance of this genetic background to VSHV and IHNV, to the birnavirus IPNV and the orthomyxovirus ISAV. Another cell line was even more refractory to the VHSV infection through different antiviral mechanisms. This collection of clonal fish and isogenic cell lines provides an interesting model to analyze the relative contribution of antiviral pathways to the resistance to different viruses.

  1. Susceptibility of Chinese Perch Brain (CPB) Cell and Mandarin Fish to Red-Spotted Grouper Nervous Necrosis Virus (RGNNV) Infection.

    Science.gov (United States)

    Tu, Jiagang; Chen, Wenjie; Fu, Xiaozhe; Lin, Qiang; Chang, Ouqin; Zhao, Lijuan; Lan, Jiangfeng; Li, Ningqiu; Lin, Li

    2016-05-19

    Nervous necrosis virus (NNV) is the causative agent of viral encephalopathy and retinopathy (VER), a neurological disease responsible for high mortality of fish species worldwide. Taking advantage of our established Chinese perch brain (CPB) cell line derived from brain tissues of Mandarin fish (Siniperca chuatsi), the susceptibility of CPB cell to Red-Spotted Grouper nervous necrosis virus (RGNNV) was evaluated. The results showed that RGNNV replicated well in CPB cells, resulting in cellular apoptosis. Moreover, the susceptibility of Mandarin fish to RGNNV was also evaluated. Abnormal swimming was observed in RGNNV-infected Mandarin fish. In addition, the cellular vacuolation and viral particles were also observed in brain tissues of RGNNV-infected Mandarin fish by Hematoxylin-eosin staining or electronic microscopy. The established RGNNV susceptible brain cell line from freshwater fish will pave a new way for the study of the pathogenicity and replication of NNV in the future.

  2. Susceptibility of Chinese Perch Brain (CPB Cell and Mandarin Fish to Red-Spotted Grouper Nervous Necrosis Virus (RGNNV Infection

    Directory of Open Access Journals (Sweden)

    Jiagang Tu

    2016-05-01

    Full Text Available Nervous necrosis virus (NNV is the causative agent of viral encephalopathy and retinopathy (VER, a neurological disease responsible for high mortality of fish species worldwide. Taking advantage of our established Chinese perch brain (CPB cell line derived from brain tissues of Mandarin fish (Siniperca chuatsi, the susceptibility of CPB cell to Red-Spotted Grouper nervous necrosis virus (RGNNV was evaluated. The results showed that RGNNV replicated well in CPB cells, resulting in cellular apoptosis. Moreover, the susceptibility of Mandarin fish to RGNNV was also evaluated. Abnormal swimming was observed in RGNNV-infected Mandarin fish. In addition, the cellular vacuolation and viral particles were also observed in brain tissues of RGNNV-infected Mandarin fish by Hematoxylin-eosin staining or electronic microscopy. The established RGNNV susceptible brain cell line from freshwater fish will pave a new way for the study of the pathogenicity and replication of NNV in the future.

  3. 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…

  4. 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.

  5. 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.

  6. 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.

  7. Impairment of mineralization by metavanadate and decavanadate solutions in a fish bone-derived cell line.

    Science.gov (United States)

    Tiago, Daniel M; Laizé, Vincent; Cancela, M Leonor; Aureliano, Manuel

    2008-06-01

    Vanadium, a trace metal known to accumulate in bone and to mimic insulin, has been shown to regulate mammalian bone formation using in vitro and in vivo systems. In the present work, short- and long-term effects of metavanadate (containing monomeric, dimeric, tetrameric and pentameric vanadate species) and decavanadate (containing decameric vanadate species) solutions on the mineralization of a fish bone-derived cell line (VSa13) were studied and compared to that of insulin. After 2 h of incubation with vanadate (10 microM in monomeric vanadate), metavanadate exhibited higher accumulation rates than decavanadate (6.85 +/- 0.40 versus 3.95 +/- 0.10 microg V/g of protein, respectively) in fish VSa13 cells and was also shown to be less toxic when applied for short periods. In longer treatments with both metavanadate and decavanadate solutions, similar effects were promoted: stimulation of cell proliferation and strong impairment (75%) of extracellular matrix (ECM) mineralization. The effect of both vanadate solutions (5 microM in monomeric vanadate), on ECM mineralization was increased in the presence of insulin (10 nM). It is concluded that chronic treatment with both vanadate solutions stimulated fish VSa13 cells proliferation and prevented ECM mineralization. Newly developed VSa13 fish cells appeared to be appropriate in the characterization of vanadate effects on vertebrate bone formation, representing a good alternative to mammalian systems.

  8. 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...

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. Inhibition of Reporter Genes by Small Interfering RNAs in Cell Culture and Living Fish

    DEFF Research Database (Denmark)

    Larashati, Sekar; Schyth, Brian Dall; Lorenzen, Niels

    2011-01-01

    as it provides low background compared to other reporter genes such as green fluorescence protein (GFP). In cell culture, the luciferase can be used as reporter gene to see the effect of gene silencing. In the living fish, the bioluminescence signal detected is influenced by the melanin pigment. Timing between...

  14. 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.

  15. 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}).

  16. 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.

  17. 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.

  18. 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...

  19. 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

  20. 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).

  1. Gene transfer and genome-wide insertional mutagenesis by retroviral transduction in fish stem cells.

    Directory of Open Access Journals (Sweden)

    Qizhi Liu

    Full Text Available Retrovirus (RV is efficient for gene transfer and integration in dividing cells of diverse organisms. RV provides a powerful tool for insertional mutagenesis (IM to identify and functionally analyze genes essential for normal and pathological processes. Here we report RV-mediated gene transfer and genome-wide IM in fish stem cells from medaka and zebrafish. Three RVs were produced for fish cell transduction: rvLegfp and rvLcherry produce green fluorescent protein (GFP and mCherry fluorescent protein respectively under control of human cytomegalovirus immediate early promoter upon any chromosomal integration, whereas rvGTgfp contains a splicing acceptor and expresses GFP only upon gene trapping (GT via intronic in-frame integration and spliced to endogenous active genes. We show that rvLegfp and rvLcherry produce a transduction efficiency of 11~23% in medaka and zebrafish stem cell lines, which is as 30~67% efficient as the positive control in NIH/3T3. Upon co-infection with rvGTgfp and rvLcherry, GFP-positive cells were much fewer than Cherry-positive cells, consistent with rareness of productive gene trapping events versus random integration. Importantly, rvGTgfp infection in the medaka haploid embryonic stem (ES cell line HX1 generated GTgfp insertion on all 24 chromosomes of the haploid genome. Similar to the mammalian haploid cells, these insertion events were presented predominantly in intergenic regions and introns but rarely in exons. RV-transduced HX1 retained the ES cell properties such as stable growth, embryoid body formation and pluripotency gene expression. Therefore, RV is proficient for gene transfer and IM in fish stem cells. Our results open new avenue for genome-wide IM in medaka haploid ES cells in culture.

  2. Dnd Is a Critical Specifier of Primordial Germ Cells in the Medaka Fish

    Directory of Open Access Journals (Sweden)

    Ni Hong

    2016-03-01

    Full Text Available Primordial germ cell (PGC specification occurs early in development. PGC specifiers have been identified in Drosophila, mouse, and human but remained elusive in most animals. Here we identify the RNA-binding protein Dnd as a critical PGC specifier in the medaka fish (Oryzias latipes. Dnd depletion specifically abolished PGCs, and its overexpression boosted PGCs. We established a single-cell culture procedure enabling lineage tracing in vitro. We show that individual blastomeres from cleavage embryos at the 32- and 64-cell stages are capable of PGC production in culture. Importantly, Dnd overexpression increases PGCs via increasing PGC precursors. Strikingly, dnd RNA forms prominent particles that segregate asymmetrically. Dnd concentrates in germ plasm and stabilizes germ plasm RNA. Therefore, Dnd is a critical specifier of fish PGCs and utilizes particle partition as a previously unidentified mechanism for asymmetric segregation. These findings offer insights into PGC specification and manipulation in medaka as a lower vertebrate model.

  3. Müller glia: Stem cells for generation and regeneration of retinal neurons in teleost fish.

    Science.gov (United States)

    Lenkowski, Jenny R; Raymond, Pamela A

    2014-05-01

    Adult zebrafish generate new neurons in the brain and retina throughout life. Growth-related neurogenesis allows a vigorous regenerative response to damage, and fish can regenerate retinal neurons, including photoreceptors, and restore functional vision following photic, chemical, or mechanical destruction of the retina. Müller glial cells in fish function as radial-glial-like neural stem cells. During adult growth, Müller glial nuclei undergo sporadic, asymmetric, self-renewing mitotic divisions in the inner nuclear layer to generate a rod progenitor that migrates along the radial fiber of the Müller glia into the outer nuclear layer, proliferates, and differentiates exclusively into rod photoreceptors. When retinal neurons are destroyed, Müller glia in the immediate vicinity of the damage partially and transiently dedifferentiate, re-express retinal progenitor and stem cell markers, re-enter the cell cycle, undergo interkinetic nuclear migration (characteristic of neuroepithelial cells), and divide once in an asymmetric, self-renewing division to generate a retinal progenitor. This daughter cell proliferates rapidly to form a compact neurogenic cluster surrounding the Müller glia; these multipotent retinal progenitors then migrate along the radial fiber to the appropriate lamina to replace missing retinal neurons. Some aspects of the injury-response in fish Müller glia resemble gliosis as observed in mammals, and mammalian Müller glia exhibit some neurogenic properties, indicative of a latent ability to regenerate retinal neurons. Understanding the specific properties of fish Müller glia that facilitate their robust capacity to generate retinal neurons will inform and inspire new clinical approaches for treating blindness and visual loss with regenerative medicine.

  4. Snakehead-fish cell line, SSN-1 (Ophicephalus striatus) as a model for cadmium genotoxicity testing.

    Science.gov (United States)

    Aramphongphan, A; Laovitthayanggoon, S; Himakoun, L

    2009-08-01

    The purpose of this study is to evaluate the potential of snakehead-fish cell line (SSN-1 cells) derived from a striped snakehead (Ophicephalus striatus) as a model in the genotoxic assessment of cadmium (Cd). The first approach employed was to determine the contaminated Cd levels in commercial snakehead fish by the Graphite Atomic Absorption Spectrophotometer. In the second approach, the sensitivity of SSN-1 cells to cytotoxicity and genotoxicity of Cd was assessed by Trypan blue and micronucleus assays following 24, 48, and 72 h of incubation period. Exposure of SSN-1 cells to four increasing Cd concentrations ranging from 0.005 to 5 ppm for 72 h did not affect their survival as compared to the control cells. The Cd uptake by SSN-1 cells showed a concentration-dependent increase in intracellular Cd levels. Three non-cytotoxic Cd concentrations (0.05, 0.5, and 5 ppm) showed a concentration-dependent genotoxic effect, compared to relevant control cells. Both micronucleus frequencies and cadmium uptake levels by SSN-1 cells depended on exposure concentrations. These results showed that the SSN-1 cells are suitable for use as a model for in vitro Cd genotoxicity testing.

  5. Optimization of Betanodavirus culture and enumeration in striped snakehead fish cells.

    Science.gov (United States)

    Hick, Paul; Tweedie, Alison; Whittington, Richard J

    2011-05-01

    An optimized culture method for detection of infection of fish with the Red spotted grouper nervous necrosis virus (RGNNV) genotype of betanodavirus in striped snakehead (SSN-1, Channa striatus) cells is described. Inoculation of fish tissue homogenates at the same time or within 4 hr of seeding the SSN-1 cells was as sensitive as the method recommended by the World Organization for Animal Health, where homogenates were adsorbed onto an established cell monolayer. Such modification halved the time required and the costs of consumables, and reduced the potential for error when processing large numbers of samples. Positive culture results were obtained from 88.3% of 392 fish tissue homogenates in which RGNNV was detected using a quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay; 99.7% of 943 tissue homogenates, which were qRT-PCR negative, were cell culture negative. Cytopathic effect (CPE) was characterized by large intracytoplasmic vacuoles in 0.1-60% of cells. Detachment of affected cells from the culture surface resulting in progressive disruption of the monolayer occurred in 46.4% of primary cultures and 96.0% of subcultures of positive samples. Identification of CPE that did not disrupt the cell monolayer increased estimates of the 50% tissue culture infective dose (TCID(50)) by 1.07-2.79 logs (95% confidence interval). The predicted mean TCID(50)/ml was 3.3 logs higher when cells were inoculated less than 36 hr after subculture at less than 80% confluence compared to cells inoculated at greater than 80% confluence and more than 36 hr after subculture (P < 0.05).

  6. A light- and electron microscopic study of primordial germ cells in the zebra fish (Danio rerio).

    Science.gov (United States)

    Deniz Koç, Nazan; Yüce, Rikap

    2012-01-01

    In sexually reproducing organisms, primordial germ cells (PGCs) give rise to the cells of the germ line, the gametes. In many animals, PGCs are set apart from somatic cells early during embryogenesis. This study explores the origin of primordial germ cells (PGCs) of the zebra fish and examines their morphology during early development (1st day-15th day). PGCs were selectively stained by the alkaline phosphatase histochemical reaction and viewed by light and electron microscopy from the time they are first detectable in the yolk sac endoderm. PGCs occurred in the subendodermal space on the syncytial periblast; differing from the surrounding endodermal cells. Later the PGCs moved to between the blastoderm and yolk sac and transferred to the dorsal mesentery where they formed gonadal anlage with mesoderm cells. PGCs were easily distinguished from somatic cells by their morphology and low electron density of their nuclei. Under light microscopy, PCGs were rounded with a distinct cytoplasmic membrane.

  7. 鱼类的胚胎干细胞%Embryonic stem cells in fish

    Institute of Scientific and Technical Information of China (English)

    洪云汉; 桂建芳; 陈松林; 邓娇蓉; ManfredSchARTL

    2003-01-01

    胚胎干细胞(ES)是未分化的细胞培养物,来自动物的早期胚胎.它们能成为稳定的细胞系和长期冻存.在适当的条件下,ES细胞能分化成各种细胞类型,包括生殖细胞.这样,ES细胞就提供了一个有效的纽带,将动物基因组的体外和体内遗传操作连系起来.ES细胞的魅力就由其在产生和分析基因敲除老鼠中显现出来.目前, ES细胞技术仅见之老鼠, 因其它脊椎动物的ES细胞的培养和建系难获成功.在鱼类,人们已做了大量的尝试.我们以青NFDA2(Oryzias latipes)作为建立鱼类ES细胞技术的模式,通过建立并应用无滋养层细胞的培养条件,获得了来自中期囊胚的ES细胞系.青NFDA2的ES细胞和老鼠的ES细胞有很多共同特征,如二倍体核型、分化潜力和形成嵌合体.因此,在鱼类建立和应用ES细胞技术是可能的.青NFDA2ES细胞的培养条件已成功地应用到其它鱼类如斑马鱼甚至海水鱼.本文旨在以青NFDA2为模式,综述获得和应用模式鱼和经济鱼ES细胞的主要进展和前景.%Embryonic stem (ES) cells are undifferentiated cell cultures derived from early developing embryos of animals. They can be propagated into stable cell lines and cryopreserved. Under appropriate conditions, ES cells can differentiate into all cell types including germ cells. Therefore, ES cells provide a unique tool linking in vitro and in vivo genetic manipulations of animal genomes. ES cells have shown their value in mice in the production and analysis of knockout animals. Because ES cells are elusive in various other vertebrate species, this ES cell technology has so far been limited to mice. Numerous attempts have been made to develop ES cell technology in fish. We used the medaka (Oryzias latipes) as a fish model for the development of ES cell technology. We have established feeder cell-free culture conditions and obtained several ES cell lines from midblastula embryos (MBE

  8. DNA vaccination in fish promotes an early chemokine-related recruitment of B cells to the muscle

    DEFF Research Database (Denmark)

    Castro, R.; Martínez-Alonso, S.; Fischer, U.

    2013-01-01

    cells that infiltrate the muscle at the site of DNA delivery in vaccinated fish and the chemokines that may be involved in their infiltration. It was observed that B lymphocytes, both IgM+ and IgT+, represent a major infiltrating cell type in fish vaccinated with a viral hemorrhagic septicemia virus...... (VHSV) DNA vaccine, whereas in control fish injected with an oil adjuvant mainly granulocytes were attracted. While IgM+ cells were the major B cell population at early time points post vaccination, IgT+ cells represented the predominant cell type later on. Among twelve chemokine genes studied...... might explain the recruitment of immune cells to the site of DNA injection. Our results suggest that B cells are involved in the initial phase of the immune response to intramuscular DNA vaccination against VHSV. This appears to be a major difference to what we know from mammalian models where T cells...

  9. gamma-Glutamyl transpeptidase as a possible marker of Sertoli cells in fish testes for studies of xenoestrogens.

    Science.gov (United States)

    Christiansen, T; Kinnberg, K; Bjerregaard, P; Korsgaard, B

    2000-01-01

    Estrogenic chemicals are known to have marked effects on the reproductive system of male fish. Finding useful markers of reproductive effects are therefore of great importance and interest. gamma-Glutamyl transpeptidase (gamma-GTP) is a possible marker of Sertoli cells in testes of fish. In the present study we have examined the relationship between the activity of gamma-GTP and the histological structure of the Sertoli cells in testes of five fish species (guppy, Poecilia reticulata; platyfish, Xiphophorus maculatus; eelpout, Zoarces viviparus; rainbow trout, Oncorhynchus mykiss; flounder, Platichthys flesus). In general we found that the more distinct the Sertoli cells the higher the activity of gamma-GTP.

  10. Enhanced propagation of fish nodaviruses in BF-2 cells persistently infected with snakehead retrovirus (SnRV).

    Science.gov (United States)

    Nishizawa, Toyohiko; Kokawa, Yuki; Wakayama, Takako; Kinoshita, Shinichi; Yoshimizu, Mamoru

    2008-03-01

    Fish nodaviruses are causative agents of viral nervous necrosis causing high mortality in cultured marine fishes around the world. The first successful isolation of fish nodavirus was made with SSN-1 cells, which are persistently infected with snakehead retrovirus (SnRV). In the present study, a BF-2 cell line persistently infected with SnRV (PI-BF-2) was established to evaluate the influence of SnRV on the production of fish nodavirus. The PI-BF-2 cells were slightly more slender than BF-2 cells, but no difference was observed in propagation rate between both cell lines. No difference was observed in production of SnRV between PI-BF-2 and SSN-1 cell lines. Although both PI-BF-2 and BF-2 cell lines showed no cytopathic effect (CPE) after inoculation of striped jack nervous necrosis virus (SJNNV) and redspotted grouper nervous necrosis virus (RGNNV), these fish nodaviruses could be amplified in BF-2 cells, and moreover, production of fish nodaviruses in the PI-BF-2 cell line was more than 40 times higher than in BF-2 cells. Thus, it was concluded that BF-2 cell permissiveness to fish nodaviruses was enhanced by persistent infection with SnRV. Furthermore, homologous cDNA to genomic RNA of SJNNV was detected from both PI-BF-2 and SSN-1 cell lines persistently infected with SnRV. The amount of nodavirus cDNA in SJNNV-inoculated PI-BF-2 cells was clearly lower than that in SJNNV-inoculated SSN-1 cells.

  11. New tools to study RNA interference to fish viruses: Fish cell lines permanently expressing siRNAs targeting the viral polymerase of viral hemorrhagic septicemia virus

    DEFF Research Database (Denmark)

    Ruiz, S.; Schyth, Brian Dall; Encinas, P.

    2009-01-01

    Previous studies have indicated that low transfection efficiency can be a major problem when gene inhibition by the use of small interfering RNAs (siRNAs) is attempted in fish cells. This may especially be true when targeting genes of viruses which are fast replicating and which can still infect ...

  12. 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.

  13. Measured and modeled toxicokinetics in cultured fish cells and application to in vitro-in vivo toxicity extrapolation.

    Science.gov (United States)

    Stadnicka-Michalak, Julita; Tanneberger, Katrin; Schirmer, Kristin; Ashauer, Roman

    2014-01-01

    Effect concentrations in the toxicity assessment of chemicals with fish and fish cells are generally based on external exposure concentrations. External concentrations as dose metrics, may, however, hamper interpretation and extrapolation of toxicological effects because it is the internal concentration that gives rise to the biological effective dose. Thus, we need to understand the relationship between the external and internal concentrations of chemicals. The objectives of this study were to: (i) elucidate the time-course of the concentration of chemicals with a wide range of physicochemical properties in the compartments of an in vitro test system, (ii) derive a predictive model for toxicokinetics in the in vitro test system, (iii) test the hypothesis that internal effect concentrations in fish (in vivo) and fish cell lines (in vitro) correlate, and (iv) develop a quantitative in vitro to in vivo toxicity extrapolation method for fish acute toxicity. To achieve these goals, time-dependent amounts of organic chemicals were measured in medium, cells (RTgill-W1) and the plastic of exposure wells. Then, the relation between uptake, elimination rate constants, and log KOW was investigated for cells in order to develop a toxicokinetic model. This model was used to predict internal effect concentrations in cells, which were compared with internal effect concentrations in fish gills predicted by a Physiologically Based Toxicokinetic model. Our model could predict concentrations of non-volatile organic chemicals with log KOW between 0.5 and 7 in cells. The correlation of the log ratio of internal effect concentrations in fish gills and the fish gill cell line with the log KOW was significant (r>0.85, p = 0.0008, F-test). This ratio can be predicted from the log KOW of the chemical (77% of variance explained), comprising a promising model to predict lethal effects on fish based on in vitro data.

  14. Discrimination of bacteriophage infected cells using locked nucleic acid fluorescent in situ hybridization (LNA-FISH).

    Science.gov (United States)

    Vilas Boas, Diana; Almeida, Carina; Sillankorva, Sanna; Nicolau, Ana; Azeredo, Joana; Azevedo, Nuno F

    2016-01-01

    Bacteriophage-host interaction studies in biofilm structures are still challenging due to the technical limitations of traditional methods. The aim of this study was to provide a direct fluorescence in situ hybridization (FISH) method based on locked nucleic acid (LNA) probes, which targets the phage replication phase, allowing the study of population dynamics during infection. Bacteriophages specific for two biofilm-forming bacteria, Pseudomonas aeruginosa and Acinetobacter, were selected. Four LNA probes were designed and optimized for phage-specific detection and for bacterial counterstaining. To validate the method, LNA-FISH counts were compared with the traditional plaque forming unit (PFU) technique. To visualize the progression of phage infection within a biofilm, colony-biofilms were formed and infected with bacteriophages. A good correlation (r = 0.707) was observed between LNA-FISH and PFU techniques. In biofilm structures, LNA-FISH provided a good discrimination of the infected cells and also allowed the assessment of the spatial distribution of infected and non-infected populations.

  15. Permissivity of fish cell lines to three Chlamydia-related bacteria: Waddlia chondrophila, Estrella lausannensis and Parachlamydia acanthamoebae.

    Science.gov (United States)

    Kebbi-Beghdadi, Carole; Batista, Claudia; Greub, Gilbert

    2011-12-01

    Epitheliocystis is an infectious disease affecting gills and skin of various freshwater and marine fishes, associated with high mortality and reduced growth of survivors. Candidatus Piscichlamydia salmonis and Clavochlamydia salmonicola have recently been identified as aetiological agents of epitheliocystis in Atlantic Salmon. In addition, several other members of the Chlamydiales order have been identified in other fish species. To clarify the pathogenicity of Chlamydia-like organisms towards fishes, we investigated the permissivity of two fish cell lines, EPC-175 (Fathead Minnow) and RTG-2 (rainbow trout) to three Chlamydia-related bacteria: Waddlia chondrophila, Parachlamydia acanthamoebae and Estrella lausannensis. Quantitative PCR and immunofluorescence demonstrated that W. chondrophila and, to a lesser extent, E. lausannensis were able to replicate in the two cell lines tested. Waddlia chondrophila multiplied rapidly in its host cell and a strong cytopathic effect was observed. During E. lausannensis infection, we observed a limited replication of the bacteria not followed by host cell lysis. Very limited replication of P. acanthamoebae was observed in both cell lines tested. Given its high infectivity and cytopathic effect towards fish cell lines, W. chondrophila represents the most interesting Chlamydia-related bacteria to be used to develop an in vivo model of epitheliocystis disease in fishes.

  16. Immunomodulatory Effectiveness of Fish Oil and omega-3 Fatty Acids in Human Non-melanoma Skin Carcinoma Cells.

    Science.gov (United States)

    Rehman, Khurram; Mohd Amin, Mohd Cairul Iqbal; Yuen, Ng Pei; Zulfakar, Mohd Hanif

    2016-01-01

    Fish oil is composed of various fatty acids among which omega-3 fatty acids are considered as most beneficial. The effects of fish oil on the activity of a topical anticancer drug, imiquimod, and the immunomodulatory activity of omega-3 fatty acids was investigated in human basal and squamous cell carcinoma cell lines. Imiquimod-fish oil mixture exhibited higher carcinoma cell growth inhibition and immunomodulatory activity than imiquimod alone, especially against squamous cell carcinoma cells. Omega-3 fatty acids exhibited growth inhibition of both basal cell and squamous cell carcinoma cell lines and modulated the immune response. Omega-3 fatty acids of fish oil serve as inducers of interleukin-10, an anti-inflammatory cytokine, and as suppressors of interleukin-6 and tumor necrosis factor-alpha, which not only depress tumor growth but also adequately control the inflammatory side effects of imiquimod. Thus, imiquimod administration with fish oil could be beneficial for inhibition of non-melanoma skin carcinoma cells but further in vivo studies are needed to understand their role in skin cancer.

  17. CHARACTERIZATION OF LACTATE DEHYDROGENASE ISOZYME PATTERN AND MORPHOLOGY OF THREE MARINE FISH CELL LINES

    Institute of Scientific and Technical Information of China (English)

    郭华荣; 张士璀; 李红岩; 童裳亮; 相建海

    2002-01-01

    Three continuous marine fish cell lines of FG (i. e., Hounder Gill) from flounder (Paralichthys olivaceus) gill, SPH (i. e. , Sea Perch Heart) from sea perch (Lateolabrax japonicus) heart and RSBF (i.e., Red Sea Bream Fin) from red sea bream (Pagrosomus major) fin, were characterized by lactate dehydrogenase (LDH) isozyme and morphological analysis. The LDH isozyme patterns of these three cell lines and their corresponding tissues of origin were investigated and compared. The results showed: (1) No difference was found in the LDH isozyme patterns of FG and flounder gill tissue. However, the LDH isozyme patterns of SPH and RSBF were significantly different from their corresponding tissues of origin; (2) LDH isozyme patterns of FG, SPH and RSBF were markedly different from each other and could serve as genetic markers for species identification and detection of cross contamination. Morphological change analysis of these three cell lines in comparison to their original tissues indicated that FG cells still appeared epithelioid without morphological transformation. However, morphological changes were found in SPH and RSBF compared to their original tissues. Therefore, the cellular morphology was still plastic in the relatively stable culture conditions, and it was possible that change of LDH patterns wasrelated to morphological changes of fish cells in vitro.

  18. CHARACTERIZATION OF LACTATE DEHYDROGENASE ISOZYME PATTERN AND MORPHOLOGY OF THREE MARINE FISH CELL LINES

    Institute of Scientific and Technical Information of China (English)

    郭华荣; 张士璀; 李红岩; 童裳亮; 相建海

    2002-01-01

    Three continuous marine fish cell lines of FG (i.e. , Flounder Gill) from flounder (Paralichthys olivaceus) gill, SPH (i.e., Sea Perch Heart) fro m sea perch (Lateolabrax japonicus) heart and RSBF (i.e., Red Sea Bream Fin) from red se a bream (Pagrosomus major) fin, were characterized by lactate dehydrogenase (LDH) is ozyme and morphological analysis. The LDH isozyme patterns of these three cell lines and their corresponding tissues of origin were investigated and compared. The results sho wed: (1) No difference was found in the LDH isozyme patterns of FG and flounder gill tissue. However, the LDH isozyme patterns of SPH and RSBF were significantly different from their cor responding tissues of origin; (2) LDH isozyme patterns of FG, SPH and RSBF were markedly di fferent from each other and could serve as genetic markers for species identification and de tection of cross contamination. Morphological change analysis of these three cell lines in compa rison to their original tissues indicated that FG cells still appeared epithelioid without mor phological transformation. However, morphological changes were found in SPH and RSBF compa red to their original tissues. Therefore, the cellular morphology was still plastic in the relatively stable culture conditions, and it was possible that change of LDH patterns was related to morphological changes of fish cells in vitro.

  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. Autophagy induced by snakehead fish vesiculovirus inhibited its replication in SSN-1 cell line.

    Science.gov (United States)

    Wang, Yao; Chen, Nan; Hegazy, Abeer M; Liu, Xiaodan; Wu, Zhixin; Liu, Xueqin; Zhao, Lijuan; Qin, Qiwei; Lan, Jiangfeng; Lin, Li

    2016-08-01

    Autophagy plays an important role in host protection against pathogen infection through activating innate and adaptive immunity. In the present study, we observed that the infection of snakehead fish vesiculovirus (SHVV) could induce apparent autophagy in striped snakehead fish cell line (SSN-1), including clear double-membrane vesicles, fluorescent punctate pattern of microtubule-associated protein 1 light chain 3B (SSN-LC3B) and the conversion of SSN-LC3B-Ⅰ to SSN-LC3B-Ⅱ. Furthermore, we verified that autophagy inhibited the replication of SHVV by assessing mRNA and protein level of nucleoprotein as well as virus titer in the supernatant. These results will shed a new light on the prevention of the infection of SHVV.

  2. Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization (FISH)

    DEFF Research Database (Denmark)

    Ronander, Elena; Bengtsson, Dominique C; Joergensen, Louise;

    2012-01-01

    and the consequence of differential binding on the clinical outcome of P. falciparum infections. Recently, the mutually exclusive transcription paradigm has been called into doubt by transcription assays based on individual P. falciparum transcript identification in single infected erythrocytic cells using RNA...... fluorescent in situ hybridization (FISH) analysis of var gene transcription by the parasite in individual nuclei of P. falciparum IE(1). Here, we present a detailed protocol for carrying out the RNA-FISH methodology for analysis of var gene transcription in single-nuclei of P. falciparum infected human...... erythrocytes. The method is based on the use of digoxigenin- and biotin- labeled antisense RNA probes using the TSA Plus Fluorescence Palette System(2) (Perkin Elmer), microscopic analyses and freshly selected P. falciparum IE. The in situ hybridization method can be used to monitor transcription...

  3. Diploidized eggs reprogram adult somatic cell nuclei to pluripotency in nuclear transfer in medaka fish (Oryzias latipes).

    Science.gov (United States)

    Bubenshchikova, Ekaterina; Kaftanovskaya, Elena; Motosugi, Nami; Fujimoto, Takafumi; Arai, Katsutoshi; Kinoshita, Masato; Hashimoto, Hisashi; Ozato, Kenjiro; Wakamatsu, Yuko

    2007-12-01

    Reprogramming of adult somatic cell nuclei to pluripotency has been unsuccessful in non-mammalian animals, primarily because of chromosomal aberrations in nuclear transplants, which are considered to be caused by asynchrony between the cell cycles of the recipient egg and donor nucleus. In order to normalize the chromosomal status, we used diploidized eggs by retention of second polar body release, instead of enucleated eggs, as recipients in nuclear transfer of primary culture cells from the caudal fin of adult green fluorescent protein gene (GFP) transgenic medaka fish (Oryzias latipes). We found that 2.7% of the reconstructed embryos grew into adults that expressed GFP in various tissues in the same pattern as in the donor fish. Moreover, these fish were diploid, fertile and capable of passing the marker gene to the next generation in Mendelian fashion. We hesitate to call these fish 'clones' because we used non-enucleated eggs as recipients; in effect, they may be chimeras consisting of cells derived from diploid recipient nuclei and donor nuclei. In either case, fish adult somatic cell nuclei were reprogrammed to pluripotency and differentiated into a variety of cell types including germ cells via the use of diploidized recipient eggs.

  4. 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

  5. Be Healthy as a Fish Educational Program at the International Institute of Molecular and Cell Biology in Warsaw, Poland.

    Science.gov (United States)

    Goś, Daria; Szymańska, Ewelina; Białek-Wyrzykowska, Urszula; Wiweger, Małgorzata; Kuźnicki, Jacek

    2016-08-01

    The purpose of the Be Healthy as a Fish educational program that is organized by the International Institute of Molecular and Cell Biology (IIMCB) in Warsaw, Poland, is to educate children about the ways in which zebrafish can be used as a model organism to help scientists understand the way the human body works. We introduce Be Healthy as a Fish workshops to children in fourth to sixth grades of primary school (9-11 years old), together with two kinds of materials under the same title: a book and a movie. We focus on the field of biology in a way that complements the children's classroom curriculum and encourages them to broaden their interests in biology in the future. The Be Healthy as a Fish educational program was inaugurated in 2014 at the Warsaw Science Festival. As of October 31, 2015, 526 primary school students participated in 27 workshops. Approximately 2000 people have received the book and nearly 1700 people have watched the movie. Be Healthy as a Fish: Origin of the Title There is a popular saying in Poland that someone is "healthy as a fish" meaning that one enjoys good health. Does this imply that fish are really that healthy? Obviously, some fish may not be healthy. Just like other animals and humans, they can and do get sick. However, this common and deceptive impression of "healthy fish" results from the fact that people hardly ever have an opportunity to observe a fish that is sick. Why does our educational program have such a possibly misleading title that may not always be true? We took advantage of this provocative title and commonly known expression and assigned to it a completely new meaning: fish can get sick, but they are important for human health. Notably, this catchy sentence intrinsically combines two keywords-health and fish-which, in our opinion, makes it a good title for a successful educational program.

  6. Negligible cytotoxicity induced by different titanium dioxide nanoparticles in fish cell lines.

    Science.gov (United States)

    Bermejo-Nogales, Azucena; Connolly, Mona; Rosenkranz, Philipp; Fernández-Cruz, María-Luisa; Navas, José M

    2017-04-01

    Titanium dioxide nanoparticles (TiO2-NPs) have a wide number of applications in cosmetic, solar and paint industries due to their photocatalyst and ultraviolet blocking properties. The continuous increase in the production of TiO2-NPs enhances the risk for this manufactured nanomaterial to enter water bodies through treated effluents or agricultural amendments. TiO2-NPs have shown very low toxicity in a number of aquatic organisms. However, there are no conclusive data about their deleterious effects and on their possible mechanisms of toxic action. At this level, in vitro cell culture systems are a useful tool to gain insight about processes underlying the toxicity of a wide variety of substances, including nanomaterials. Differences in the physiology of different taxa make advisable the use of cells coming from the taxon of interest, but collecting data from a variety of cellular types allows a better understanding of the studied processes. Taking all this into account, the aim of the present study was to assess the toxicity of three types of TiO2-NP, rutile hydrophobic (NM-103), rutile hydrophilic (NM-104) and rutile-anatase (NM-105), obtained from the EU Joint Research Centre (JRC) repository, using various fish cell lines (RTG-2, PLHC-1, RTH-149, RTL-W1) and rainbow trout primary hepatocytes. For comparative purposes, the effect of different dispersion protocols, end-point assays and extended exposure time was studied in a fish cell line (RTG-2) and in the rat hepatoma cell line (H4IIE). TiO2-NPs dispersions showed a variable degree of aggregation in cell culture media. Disruption of mitochondrial metabolic activity, plasma membrane integrity and lysosome function was not detected in any cell line after exposure to TiO2-NPs at any time and concentration ranges tested. These results are indicative of a low toxicity of the TiO2-NPs tested and show the usefulness of fish cells maintained in vitro as high throughput screening methods that can facilitate further

  7. Visualization and quantitative analysis of extrachromosomal telomere-repeat DNA in individual human cells by Halo-FISH.

    Science.gov (United States)

    Komosa, Martin; Root, Heather; Meyn, M Stephen

    2015-02-27

    Current methods for characterizing extrachromosomal nuclear DNA in mammalian cells do not permit single-cell analysis, are often semi-quantitative and frequently biased toward the detection of circular species. To overcome these limitations, we developed Halo-FISH to visualize and quantitatively analyze extrachromosomal DNA in single cells. We demonstrate Halo-FISH by using it to analyze extrachromosomal telomere-repeat (ECTR) in human cells that use the Alternative Lengthening of Telomeres (ALT) pathway(s) to maintain telomere lengths. We find that GM847 and VA13 ALT cells average ∼80 detectable G/C-strand ECTR DNA molecules/nucleus, while U2OS ALT cells average ∼18 molecules/nucleus. In comparison, human primary and telomerase-positive cells contain 300), range widely in length (200 kb) and are composed of primarily G- or C-strand telomere-repeat DNA. Halo-FISH enables, for the first time, the simultaneous analysis of ECTR DNA and chromosomal telomeres in a single cell. We find that ECTR DNA comprises ∼15% of telomere-repeat DNA in GM847 and VA13 cells, but FISH can facilitate the study of a wide variety of extrachromosomal DNA in mammalian cells.

  8. Toxicity and cytopathogenic properties toward human melanoma cells of activated cancer therapeutics in zebra fish.

    Science.gov (United States)

    Lewis, Thomas J

    2010-03-01

    There is an increasing body of data showing that activated cancer therapy--the synergistic effect of "preloaded" molecules and a tuned energy source to produce cytopathogenic moieties--is a promising new modality for cancer treatment. The key activated therapies are photodynamic therapy (PDT), which involves the synergy between light and photosensitizer molecules, and ultrasound activated therapy (USAT; also referred to as sonodynamic therapy), which involves the synergy between ultrasound and sonosensitizer molecules. PDT is a well-known activated therapy with roots dating back to 1900. However, minimal data exist on USAT. One reason is the lack of suitable sonosensitizers for clinical USAT use. The authors present both LC(50) toxicity and cancer cell cytotoxicity studies on 2 dual activation agents. These compounds function as both sonosensitizers and photosensitizers, and are referred to as SonneLux agents, designated SF1 and SF2. The sensitizers are derived from chlorophyll and are metal centered porphyrins known to specifically accumulate in hyperproliferating tissue. LC(50) studies on both SF1 and SF2 as determined in zebra fish reveal that both are essentially nontoxic to zebra fish. In the worst case, 5% zebra fish death is noted at the maximum soluble concentration of the sensitizer. In the cytotoxicity studies, melanoma cell line WM-266-4, derived from a metastatic site of a malignant melanoma, was tested against SF1 and SF2. Both sensitizer systems showed marked efficacy in the destruction of the implanted melanoma cells. They show great promise for clinical use in the future.

  9. Computational Graph Model for 3D Cells Tracking in Zebra Fish Datasets

    Science.gov (United States)

    Zhang, Lelin; Xiong, Hongkai; Zhao, Yang; Zhang, Kai; Zhou, Xiaobo

    2007-11-01

    This paper leads to a novel technique for tracking and identification of zebra-fish cells in 3D image sequences, extending graph-based multi-objects tracking algorithm to 3D applications. As raised in previous work of 2D graph-based method, separated cells are modeled as vertices that connected by edges. Then the tracking work is simplified to that of vertices matching between graphs generated from consecutive frames. Graph-based tracking is composed of three steps: graph generation, initial source vertices selection and graph saturation. To satisfy demands in this work separated cell records are segmented from original datasets using 3D level-set algorithms. Besides, advancements are achieved in each of the step including graph regulations, multi restrictions on source vertices and enhanced flow quantifications. Those strategies make a good compensation for graph-based multi-objects tracking method in 2D space. Experiments are carried out in 3D datasets sampled from zebra fish, results of which shows that this enhanced method could be potentially applied to tracking of objects with diverse features.

  10. Experimental improvements in combining CARD-FISH and flow cytometry for bacterial cell quantification.

    Science.gov (United States)

    Manti, Anita; Boi, Paola; Amalfitano, Stefano; Puddu, Alberto; Papa, Stefano

    2011-12-01

    Flow cytometry and Fluorescence In Situ Hybridization are common methods of identifying and quantifying bacterial cells. The combination of cytometric rapidity and multi-parametric accuracy with the phylogenetic specificity of oligonucleotide FISH probes has been regarded as a powerful and emerging tool in aquatic microbiology. In the present work, tests were carried out on E. coli pure culture and marine bacteria using an in-solution hybridization protocol revealing high efficiency hybridization signal for the first one and a lower for the second one. Other experiments were conducted on natural samples following the established CARD-FISH protocol on filter performed in a closed system, with the aim of improving cell detachment and detection. The hybridized cells were then subsequently re-suspended from the membrane filters by means of an optimized detachment procedure. The cytometric enumeration of hybridized marine bacteria reached 85.7%±18.1% of total events. The quality of the cytograms suggests that the procedures described may be applicable to the cytometric quantification of phylogenetic groups within natural microbial communities.

  11. COMPARATIVE PRODUCTION OF SINGLE CELL PROTEIN FROM FISH PROTEIN ISOLATE WASTAGE AND ULTRA FILTERED CHEESE WHEY

    Directory of Open Access Journals (Sweden)

    Soroush Haghighi-Manesh

    2013-02-01

    Full Text Available Fish protein isolate wastage and ultra filtered cheese whey were used as substrates for fermentation by Kluyveromyces marxianus to produce single cell protein, under batch and aerobic condition in which pH and temperature were adjusted to 4.5 and 35°C. The produced biomass was analyzed for protein content in different periods of time during fermentation. About 82% and 75% of total protein was produced in the first 18 h of 96 h fermentation of ultra filtered cheese whey and protein isolate wastage respectively, which can be an indication of the exponential phase of the yeast growth. The results of biomass yield measurements during 96 h process also confirm this finding. Moreover, since ultra filtered cheese whey was higher in single cell protein yield, solubility, water holding capacity, water absorption and power of biological and chemical oxygen demand reduction, and also was lower in foam overrun and stability than fish protein isolate wastage, it was selected as the suitable substrate for single cell protein production.

  12. Immunocytochemical identification and localization of APUD cells in the gut of seven stomachless teleost fishes

    Institute of Scientific and Technical Information of China (English)

    Qian Sheng Pan; Zhi Ping Fang; Ya Xin Zhao

    2000-01-01

    AIM To study the cell types, localization,distribution density and morphology of APUD cells in the intestinal mucosa of stomachless teleost fishes.METHOD By using the peroxidaseantiperoxidase complex ( PAP )immunocytochemical staining technique the identification, localization and morphology of immunoreactive (IR) endocrine cells seattered in the intestinal mucosa of grass carp ( Cyenopharyngodon idellus ), black carp (Mylopharyngodon piceus ) and common carp (Cyprinus carpio ) were investigated with 20 kinds of antisera prepared against mammalian peptide hormones of APUD cells, and likewise by using avidin-biotin-peroxidase complex (ABC)method those of silver carp ( Hypophthalmichthys molitrix ), bighead (Aristichthys nobilis ), silver crucian carp (Carassius gibelio ) and bluntnose black bream ( Megalobrama amblyocephala ) were also studied with 5 different antisera. The replacement of the first antiserum by phosphate buffered saline (PBS) was employed as a control. IR endocrine cells were counted with a square-mesh ocular micrometer from 10 fields selected randomly in every section of each part of the intestine specimen. The average number of IR endocrine cells per mm2 was counted to quantify their distribution density.RESULT Gastrin (GAS)-, Gastric inhibitory peptide (GIP)-, glucagon (GLU)-, glucagon-like immunoreactants ( GLI )-, bovine pancreatic polypeptide (BPP)-, leucine-enkephalin (ENK)-and substance P (SP)-IR endocrine cells were found in the gut of grass carp, black carp and common carp, and somatostatin ( SOM )-IR endocrine cells were only seen in common carp.GAS-, GIP- and GLU-IR endocrine cells were found in the intestinal mucosa of silver carp,bighead, silver crucian carp and bluntnose black bream. Most of IR endocrine cells had the higher distribution density in the foregut and midgut,and were longer in shape. They had a long apical cytoplasmic process extended to the gut lumen and a basal process extended to adjacent cells or basement membrane and

  13. 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.

  14. Fish liver and gill cells as cytogenotoxic indicators in assessment of water quality.

    Science.gov (United States)

    Kaur, Rajbir; Dua, Anish

    2016-09-01

    The present study was aimed to evaluate the effect of municipal wastewater of Tung Dhab drain to freshwater fish Labeo rohita through genotoxicity assays. The idea is to explore and standardize procedures for assessment of toxic impact of pollutant at the cellular level in aquatic species. Hence, chronic, non-renewal toxicity tests were performed with sublethal concentrations (17.7, 26.6 and 35.4 %) of wastewater for exposure durations of 15, 30 and 60 days. Recovery experiments were also performed for duration of 60 days. Higher micronuclei (MN) and binuclei (BN) frequencies were recorded in fluorescently stained liver cells as compared to frequencies obtained for gill cells and erythrocytes. The induced frequencies of MN, BN and nuclear abnormalities (NA) were found in the following order: MN (liver cells) > BN (liver cells) > BN (gill cells) > MN (gill cells) > NA (erythrocytes) > MN (erythrocytes). The results pointed towards the mutagenic nature of water quality of Tung Dhab drain and recommend the incorporation of the liver and gill cells as biomarkers of cytotoxicity over erythrocytes in chronic experiments during water monitoring programs.

  15. Reduction of splenic immunosuppressive cells and enhancement of anti-tumor immunity by synergy of fish oil and selenium yeast.

    Directory of Open Access Journals (Sweden)

    Hang Wang

    Full Text Available Growing evidence has shown that regulatory T cells (Tregs and myeloid-derived suppressor cells (MDSCs abnormally increase in cancer cachectic patients. Suppressions of Tregs and MDSCs may enhance anti-tumor immunity for cancer patients. Fish oil and selenium have been known to have many biological activities such as anti-inflammation and anti-oxidation. Whether fish oil and/or selenium have an additional effect on population of immunosuppressive cells in tumor-bearing hosts remained elusive and controversial. To gain insights into their roles on anti-tumor immunity, we studied the fish oil- and/or selenium-mediated tumor suppression and immunity on lung carcinoma, whereof cachexia develops. Advancement of cachexia in a murine lung cancer model manifested with such indicative symptoms as weight loss, chronic inflammation and disturbed immune functionality. The elevation of Tregs and MDSCs in spleens of tumor-bearing mice was positively correlated with tumor burdens. Consumption of either fish oil or selenium had little or no effect on the levels of Tregs and MDSCs. However, consumption of both fish oil and selenium together presented a synergistic effect--the population of Tregs and MDSCs decreased as opposed to increase of anti-tumor immunity when both fish oil and selenium were supplemented simultaneously, whereby losses of body weight and muscle/fat mass were alleviated significantly.

  16. 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.

  17. 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.

  18. Correlation-induced network oscillations of pyramidal cells in the weakly electric fish - theory and experiment

    Science.gov (United States)

    Lindner, Benjamin; Doiron, Brent; Longtin, Andre; Maler, Leonard; Bastian, Joseph

    2004-03-01

    The spiking activity of pyramidal cells in the weakly electric fish is studied. It is experimentally shown that the oscillatory spiking activity of these cells increases with the spatial correlations of external stochastic input. A model network of integrate-and-fire (IF) neurons with delayed inhibitory feedback reproduces this effect. Moreover, a novel analytical approach for stochastic neuron models with weak feedback is presented that leads to a simple expression for the power spectrum of the spike train of a single neuron. The analytical results agree well with simulation results of the leaky IF neurons; they also show the same qualitative features as the experimental spectra and are helpful in understanding the deeper origin of the correlation-induced oscillations.

  19. Several synthetic progestins disrupt the glial cell specific-brain aromatase expression in developing zebra fish.

    Science.gov (United States)

    Cano-Nicolau, Joel; Garoche, Clémentine; Hinfray, Nathalie; Pellegrini, Elisabeth; Boujrad, Noureddine; Pakdel, Farzad; Kah, Olivier; Brion, François

    2016-08-15

    The effects of some progestins on fish reproduction have been recently reported revealing the hazard of this class of steroidal pharmaceuticals. However, their effects at the central nervous system level have been poorly studied until now. Notwithstanding, progesterone, although still widely considered primarily a sex hormone, is an important agent affecting many central nervous system functions. Herein, we investigated the effects of a large set of synthetic ligands of the nuclear progesterone receptor on the glial-specific expression of the zebrafish brain aromatase (cyp19a1b) using zebrafish mechanism-based assays. Progesterone and 24 progestins were first screened on transgenic cyp19a1b-GFP zebrafish embryos. We showed that progesterone, dydrogesterone, drospirenone and all the progesterone-derived progestins had no effect on GFP expression. Conversely, all progestins derived from 19-nortesterone induced GFP in a concentration-dependent manner with EC50 ranging from the low nM range to hundreds nM. The 19-nortestosterone derived progestins levonorgestrel (LNG) and norethindrone (NET) were further tested in a radial glial cell context using U251-MG cells co-transfected with zebrafish ER subtypes (zfERα, zfERβ1 or zfERβ2) and cyp19a1b promoter linked to luciferase. Progesterone had no effect on luciferase activity while NET and LNG induced luciferase activity that was blocked by ICI 182,780. Zebrafish-ERs competition assays showed that NET and LNG were unable to bind to ERs, suggesting that the effects of these compounds on cyp19a1b require metabolic activation prior to elicit estrogenic activity. Overall, we demonstrate that 19-nortestosterone derived progestins elicit estrogenic activity by inducing cyp19a1b expression in radial glial cells. Given the crucial role of radial glial cells and neuro-estrogens in early development of brain, the consequences of exposure of fish to these compounds require further investigation.

  20. 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

  1. Interactive effects of metals as measured in cytotoxicity assays with established fish cell lines

    Energy Technology Data Exchange (ETDEWEB)

    Segner, H.; Schuurmann, G. [Centre for Environmental Research, Leipzig (Germany). Dept. of Chemical Ecotoxicology

    1995-12-31

    The environmental toxicity of chemicals is often judged on the basis of toxicity tests with single compounds. One major drawback of this approach is the fact, that mixture effects occurring in aquatic ecosystems with a multitude of different chemicals are not accounted for. The present work explores the use of cytotoxicity assays with established fish cell lines as a rapid and economic approach to derive basic data on joint toxicity effects of heavy metals. For the assessment of mixture toxicity, concentration addition is taken as the reference model of no interaction, and both isobolographic analysis and calculation of mixture toxicity indices are used to analyze the effect profile of various equitoxic compound mixtures. Cytotoxic endpoints used include neutral red uptake inhibition assay as a measure of cell viability, proliferation measurements to estimate toxic effects on cell growth, and analysis of glutathion contents to estimate metabolic stress effects. The single toxicity of the metals silver, mercury, cadmium, copper, zinc, lead and nickel towards the cell lines RI from rainbow trout liver and RTG-2 from rainbow trout gonads was found to depend on the chemical softness parameter of the cations. The joint effect profile will be discussed in terms of the single effects and softness domain of the heavy metals.

  2. 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

  3. Genotoxicity of tetrodotoxin from puffer fish tested in root meristem cells of Allium cepa L.

    Science.gov (United States)

    Khora, S S; Panda, K K; Panda, B B

    1997-07-01

    Tetrodotoxin (TTX) extracted and purified from puffer fish Arothron nigropunctatus was tested for genotoxicity employing the root meristem cells of Allium cepa as the assay system. The genotoxicity endpoints investigated were mitotic index (MI), meta-anaphases with spindle aberrations, interphases with micronuclei (MNC) and sister chromatid exchanges (SCE) in metaphase chromosomes. The results demonstrated that TTX inhibited mitosis at concentrations of > or = 30 microM as evident by the fall of MI, but failed to induce MNC at significant levels at any of the concentrations tested (10-100 microM). TTX was thus proved to be neither clastogenic nor aneugenic in the present study. It was, however, noteworthy that TTX at far lower concentrations, 0.1-5.0 microM, significantly enhanced the frequencies of SCE which indicated possible interference of the toxin in DNA replication and repair.

  4. 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.

  5. Identification of antiviralrelevant genes in the cultured fish cells induced by UV-inactivated virus

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    UV-inactivated grass carp hemorrhage virus (GCHV) can induce high titer of interferon in cultured CAB (crucian carp (Carassius auratus L.) blastulae) cells, and thus defend host cells against the virus invasion. The mechanism is proposed that an antiviral state should be established in the host cells by activating expression of a set of antivi-ral-relevant genes. In this study, suppressive subtractive hy-bridization is applied to constructing a subtracted cDNA library with mRNAs isolated from UV-inactivated GCHV infected and mock-infected CAB cells. 272 differential cDNA fragments are identified by both PCR and dot blot from the subtractive cDNA library. Sequencing analysis reveals 69 genes, including 46 known gene homologues, and 23 unknown putative genes. The known genes include the genes involved in interferon signaling pathways, such as Stat1 and Jak1, the antiviral genes, such as Mx and Viperin, and a set of interferon-stimulated genes observed in mammalian cells. Most of the unknown putative genes contain AU-rich ele-ment in their sequences. Differential expressions of these genes are further confirmed by virtual Northern blot and RT-PCR. The data imply that UV-inactivated GCHV is not only able to induce production of interferon in the infected CAB cells, but also leads to the expression of a series of antiviral-relevant genes or immune-relevant genes, and therefore reveals that the signaling pathway of interferon system and antiviral mechanism in fish are similar to those in mammals.

  6. Colon cancer cell chemosensitisation by fish oil emulsion involves apoptotic mitochondria pathway.

    Science.gov (United States)

    Granci, Virginie; Cai, Fang; Lecumberri, Elena; Clerc, Aurélie; Dupertuis, Yves M; Pichard, Claude

    2013-04-14

    Adjuvant use of safe compounds with anti-tumour properties has been proposed to improve cancer chemotherapy outcome. We aimed to investigate the effects of fish oil emulsion (FOE) rich in n-3 PUFA with the standard chemotherapeutic agents 5-fluorouracil (5-FU), oxaliplatin (OX) or irinotecan (IRI) on two human colorectal adenocarcinoma cells with different genetic backgrounds. The HT-29 (Bax+/+) and LS174T (Bax-/-) cells were co-treated for 24-72 h with 1 μm-5-FU, 1 μm-OX or 10 μm-IRI and/or FOE dilution corresponding to 24 μm-EPA and 20·5 μm-DHA. Soyabean oil emulsion (SOE) was used as isoenergetic and isolipid control. Cell viability, apoptosis and nuclear morphological changes were evaluated by cytotoxic colorimetric assay, flow cytometry analysis with annexin V and 4',6'-diamidino-2-phenylindole staining, respectively. A cationic fluorescent probe was used to evaluate mitochondrial dysfunction, and protein expression involved in mitochondrial apoptosis was determined by Western blot. In contrast to SOE, co-treatment with FOE enhanced significantly the pro-apoptotic and cytotoxic effects of 5-FU, OX or IRI in HT-29 but not in LS174T cells (two-way ANOVA, P <0.01). These results were confirmed by the formation of apoptotic bodies in HT-29 cells. A significant increase in mitochondrial membrane depolarisation was observed after the combination of 5-FU or IRI with FOE in HT-29 but not in LS174T cells (P <0.05). Co-administration of FOE with the standard agents, 5-FU, OX and IRI, could be a good alternative to increase the efficacy of chemotherapeutic protocols through a Bax-dependent mitochondrial pathway.

  7. Identification of Barramundi (Lates calcarifer) DC-SCRIPT, a Specific Molecular Marker for Dendritic Cells in Fish.

    Science.gov (United States)

    Zoccola, Emmanuelle; Delamare-Deboutteville, Jérôme; Barnes, Andrew C

    2015-01-01

    Antigen presentation is a critical step bridging innate immune recognition and specific immune memory. In mammals, the process is orchestrated by dendritic cells (DCs) in the lymphatic system, which initiate clonal proliferation of antigen-specific lymphocytes. However, fish lack a classical lymphatic system and there are currently no cellular markers for DCs in fish, thus antigen-presentation in fish is poorly understood. Recently, antigen-presenting cells similar in structure and function to mammalian DCs were identified in various fish, including rainbow trout (Oncorhynchus mykiss) and zebrafish (Danio rerio). The present study aimed to identify a potential molecular marker for DCs in fish and therefore targeted DC-SCRIPT, a well-conserved zinc finger protein that is preferentially expressed in all sub-types of human DCs. Putative dendritic cells were obtained in culture by maturation of spleen and pronephros-derived monocytes. DC-SCRIPT was identified in barramundi by homology using RACE PCR and genome walking. Specific expression of DC-SCRIPT was detected in barramundi cells by Stellaris mRNA FISH, in combination with MHCII expression when exposed to bacterial derived peptidoglycan, suggesting the presence of DCs in L. calcarifer. Moreover, morphological identification was achieved by light microscopy of cytospins prepared from these cultures. The cultured cells were morphologically similar to mammalian and trout DCs. Migration assays determined that these cells have the ability to move towards pathogens and pathogen associated molecular patterns, with a preference for peptidoglycans over lipopolysaccharides. The cells were also strongly phagocytic, engulfing bacteria and rapidly breaking them down. Barramundi DCs induced significant proliferation of responder populations of T-lymphocytes, supporting their role as antigen presenting cells. DC-SCRIPT expression in head kidney was higher 6 and 24 h following intraperitoneal challenge with peptidoglycan and

  8. Role of dietary fish oil on nitric oxide synthase activity and oxidative status in mice red blood cells.

    Science.gov (United States)

    Martins, Marcela A; Moss, Monique B; Mendes, Iara K S; Águila, Márcia B; Mandarim-de-Lacerda, Carlos Alberto; Brunini, Tatiana M C; Mendes-Ribeiro, Antônio Cláudio

    2014-12-01

    The consumption of n-3 polyunsaturated fatty acids (PUFAs) derived from fish oil concomitant with a reduced intake of saturated fats is associated with cardiovascular benefits, which may result from the participation of nitric oxide (NO). In contrast, PUFAs are vulnerable to peroxidation, which could affect the oxidative stability of the cell and reduce NO bioavailability. Therefore, we investigated the effects of high fat diets with increasing amounts of fish oil (0-40% of energy) in place of lard on the l-arginine-NO pathway, the arginase pathway and oxidative status in mice red blood cells (RBC). We found that l-arginine transport, as well as NO synthase (NOS) expression and activity, was enhanced by the highest doses of fish oil (30 and 40%). In contrast, diets rich in lard led to NOS expression and activity impairment. Arginase expression was not significantly affected by any of the dietary regimens. No significant difference in protein and lipid oxidative markers was observed among any of the fish-oil fed mice; only lard feeding induced protein damage in addition to a decreased superoxide dismutase activity. These data suggest that a substantial dose of fish oil, but not low doses, activates the RBC l-arginine-NO pathway without resulting in oxidative damage.

  9. Meat and fish consumption and the risk of renal cell carcinoma in the European prospective investigation into cancer and nutrition

    NARCIS (Netherlands)

    Rohrmann, Sabine; Linseisen, Jakob; Overvad, Kim; Wurtz, Anne Mette Lund; Roswall, Nina; Tjonneland, Anne; Boutron-Ruault, Marie-Christine; Racine, Antoine; Bastide, Nadia; Palli, Domenico; Agnoli, Claudia; Panico, Salvatore; Tumino, Rosario; Sacerdote, Carlotta; Weikert, Steffen; Steffen, Annika; Kuehn, Tilman; Li, Kuanrong; Khaw, Kay-Tee; Wareham, Nicholas J.; Bradbury, Kathryn E.; Peppa, Eleni; Trichopoulou, Antonia; Trichopoulos, Dimitrios; Bueno-de-Mesquita, H. Bas; Peeters, Petra H. M.; Hjartaker, Anette; Skeie, Guri; Weiderpass, Elisabete; Jakszyn, Paula; Dorronsoro, Miren; Barricarte, Aurelio; Santiuste de Pablos, Carmen; Molina-Montes, Esther; Alonso de la Torre, Ramon; Ericson, Ulrika; Sonestedt, Emily; Johansson, Mattias; Ljungberg, Borje; Freisling, Heinz; Romieu, Isabelle; Cross, Amanda J.; Vergnaud, Anne-Claire; Riboli, Elio; Boeing, Heiner

    2015-01-01

    Renal cell cancer (RCC) incidence varies worldwide with a higher incidence in developed countries and lifestyle is likely to contribute to the development of this disease. We examined whether meat and fish consumption were related to the risk of RCC in the European Prospective Investigation into Can

  10. Germ cell transplantation using sexually competent fish: an approach for rapid propagation of endangered and valuable germlines.

    Directory of Open Access Journals (Sweden)

    Sullip K Majhi

    Full Text Available The transplantation of germ cells into adult recipient gonads is a tool with wide applications in animal breeding and conservation of valuable and/or endangered species; it also provides a means for basic studies involving germ cell (GC proliferation and differentiation. Here we describe the establishment of a working model for xenogeneic germ cell transplantation (GCT in sexually competent fish. Spermatogonial cells isolated from juveniles of one species, the pejerrey Odontesthes bonariensis (Atherinopsidae, were surgically transplanted into the gonads of sexually mature Patagonian pejerrey O. hatcheri, which have been partially depleted of endogenous GCs by a combination of Busulfan (40 mg/kg and high water temperature (25 degrees C treatments. The observation of the donor cells' behavior showed that transplanted spermatogonial cells were able to recolonize the recipients' gonads and resume spermatogenesis within 6 months from the GCT. The presence of donor-derived gametes was confirmed by PCR in 20% of the surrogate O. hatcheri fathers at 6 months and crosses with O. bonariensis mothers produced hybrids and pure O. bonariensis, with donor-derived germline transmission rates of 1.2-13.3%. These findings indicate that transplantation of spermatogonial cells into sexually competent fish can shorten considerably the production time of donor-derived gametes and offspring and could play a vital role in germline conservation and propagation of valued and/or endangered fish species.

  11. 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…

  12. In vitro characterization of 6-Coumarin loaded solid lipid nanoparticles and their uptake by immunocompetent fish cells.

    Science.gov (United States)

    Trapani, Adriana; Mandracchia, Delia; Di Franco, Cinzia; Cordero, Héctor; Morcillo, Patricia; Comparelli, Roberto; Cuesta, Alberto; Esteban, Maria Angeles

    2015-03-01

    The primary aim of the present work was to evaluate the in vitro uptake of 6-Coumarin (6COUM) loaded solid lipid nanoparticles (SLN) by two gilthead seabream (Sparus aurata L.) cell types: an established cell line (SAF-1 cells) and the primary cultures of head-kidney (HK)-the main haemopoietic organ in fish, equivalent to mammalian bone marrow-leucocytes. For this purpose, after the physicochemical characterization of SLN, the uptake by those immunocompetent fish cells was evaluated using flow cytometry and confocal microscopy. Concomitantly, the uptake of 6-COUM loaded SLN was compared with that achieved with 6-COUM loaded pectin microparticles (MPs), which were selected as a competitor of the delivery carriers. After SLN and MP physicochemical characterization, the results demonstrated that SAF-1 cells were able to internalize high percentages of 6-COUM SLNs when incubated for 4, 8 and 24h, with the highest SLN concentration tested (10 μg/ml). The ability of HK leucocytes to internalize SLN was also found to vary depending on both incubation time and SLN concentration. The highest values of HK leucocytes internalizing SLN particles (around 16%) were detected at the maximum SLN concentration (20 μg/ml) at incubation times of 4 or 8h. Conversely, HK leucocytes were unable to internalize MPs at any tested concentration and incubation time. A possible mechanism explaining the uptake into cells is proposed. The present work constitutes the first approximation to consider SLN as nanocarriers for delivering biologically active substances to fish.

  13. Overexpression a novel zebra fish spermatogenesis-associated gene 17 (SPATA17) induces apoptosis in GC-1 cells.

    Science.gov (United States)

    Nie, Dongsong; Liu, Y; Xiang, Y

    2011-08-01

    The spermatogenesis-associated 17 gene (SPATA17, previously named MSRG-11) was reported to be a candidate spermatocyte apoptosis-related gene which may play a critical role in human spermatogenesis, especially in meiosis. Analysis of SPATA17 expression and regulation in zebra fish may provide insight into the understanding of the complicated process of gonadogenesis and its potential function in spermatocyte cell apoptosis. In this study, we cloned and characterized the SPATA17 gene from zebra fish which consists of nine exons separated by eight introns. The consensus open reading frame (1258 bp) encodes a polypeptide of 357 amino acids which shares 44% identity with the human SPATA17 gene. Bioinformatic analysis reveals that SPATA17 protein contains three short calmodulin-binding motifs (IQ motif) and is considered to play a critical role in interactions with CaM proteins. Multi-tissue RT-PCR and Northern blot results demonstrated that the zebra fish SPATA17 gene was expressed strongly in testis and a slight amount of expression in ovary. Flow cytometry analysis and genomic DNA ladders result showed that the expression of SPATA17 protein in the GC-1 cell line could accelerate cell apoptosis. Analysis of the SPATA17 sequence and its spatial expression pattern indicate that this gene is highly conserved and may play an important role in the process of zebra fish gonadogenesis.

  14. Salmonid alphavirus replicon is functional in fish, mammalian and insect cells and in vivo in shrimps (Litopenaeus vannamei).

    Science.gov (United States)

    Olsen, Christel M; Pemula, Anand Kumar; Braaen, Stine; Sankaran, Krishnan; Rimstad, Espen

    2013-11-19

    The Salmonid alphavirus (SAV) is the etiological agent of pancreas disease in Atlantic salmon (Salmo salar) and Sleeping disease in rainbow trout (Oncorhynchus mykiss). SAV differs from alphaviruses infecting terrestrial animals in that it infects salmonid fish at low temperatures and does not use an arthropod vector for transmission. In this study we have shown that a SAVbased replicon could express proteins when driven by the subgenomic promoter in vitro in cells from fish, mammals and insects, as well as in vivo in shrimps (Litopanaeus vannamei). The SAV-replicon was found to be functional at temperatures ranging from 4 to 37°C. Protein expression was slow and moderate compared to that reported from terrestrial alphavirus replicons or from vectors where protein expression was under control of the immediate early CMV-promoter. No cytopathic effect was visually observable in cells transfected with SAV-replicon vectors. Double stranded RNA was present for several days after transfection of the SAV-replicon in fish cell lines and its presence was indicated also in shrimp. The combination of prolonged dsRNA production, low toxicity, and wide temperature range for expression, may potentially be advantageous for the use of the SAV replicon to induce immune responses in aquaculture of fish and shrimp.

  15. Cytotoxicity of cuprous oxide nanoparticles to fish blood cells: hemolysis and internalization

    Energy Technology Data Exchange (ETDEWEB)

    Chen Liqiang, E-mail: chenlq@ynu.edu.cn; Kang Bin [Yunnan University, Asian International Rivers Center, Yunnan Key Laboratory of International Rivers and Trans-boundary Eco-security (China); Ling Jian [Yunnan University, College of Chemistry and Chemical Engineering (China)

    2013-03-15

    Cuprous oxide nanoparticles (Cu{sub 2}O NPs) possess unique physical and chemical properties which are employed in a broad variety of applications. However, little is known about the adverse effects of Cu{sub 2}O NPs on organisms. In the current study, in vitro cytotoxicity of Cu{sub 2}O NPs (ca. 60 nm in diameter) to the blood cells of freshwater fish Carassius auratus was evaluated. A concentration-dependent hemolytic activity of Cu{sub 2}O NPs to red blood cells (RBCs) and the phagocytosis of Cu{sub 2}O NPs by leukocytes were revealed. The results showed that dosages of Cu{sub 2}O NPs greater than 40 {mu}g/mL were toxic to blood cells, and could cause serious membrane damage to RBCs. The EC{sub 50} value of Cu{sub 2}O NPs as obtained from RBCs and whole blood exposure was 26 and 63 {mu}g/mL, respectively. The generation of reactive oxygen species and the direct interaction between Cu{sub 2}O NPs and the cell membrane were suggested as the possible mechanism for cytotoxicity, and the intrinsic hemolytic active of Cu{sub 2}O NPs was the main contributor to the toxicity rather than solubilized copper ions. The adsorption of plasma proteins on the surfaces of Cu{sub 2}O NPs led to their aggregation in whole blood, and aggregate formation can significantly alleviate the hemolytic effect and subsequently mediate the phagocytosis of Cu{sub 2}O NPs by leukocytes.

  16. Glucocorticoid-mediated potentiation of cytochrome P4501A induction in a fish hepatoma cell line

    Energy Technology Data Exchange (ETDEWEB)

    Celander, M.; Hahn, M.E.; Stegeman, J.J. [Woods Hole Oceanographic Institution, MA (United States)

    1995-12-31

    Induction of cytochrome P4501A (CYP1A) is widely used as a biomarker of exposure to polycyclic aromatic hydrocarbons (PAH) and planar halogenated aromatic hydrocarbons (PHAH). Increasingly, potency of CYP1A inducers in fish is being determined in cells in culture including the Poeciliopsis lucida hepatocellular carcinoma cell line (PLHC-1). The authors used this cell line to investigate the effects of various glucocorticoid receptor (GR) agonists on that response. CYP1A in PLHC-1 cultures was highly responsive to treatment with PAH- or PHAH-type inducers including B-naphthoflavone (BNF), 3,3{prime},4,4{prime}-tetrachlorobiphenyl (TCB) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). TCDD was found to be three orders of magnitude more potent than BNF or TCB as an inducer of the CYP1A activity ethoxyresorufin-O-deethylase (EROD). The apparent efficacy for the induction by BNF was 50% and by TCB 10% of that obtained with TCDD. Induction of CYP1A was potentiated when these aryl hydrocarbon receptor (AHR) agonists were co-administered with GR agonists, such as dexamethasone D(EX), cortisol or prednisone. The magnitude of the potentiation of CYP1A induction varied with the different GR- and AHR agonists tested, but also with doses of AHR agonists and duration of exposure. Thus, highest degree of potentiation of CYP1A (up to 20-fold) in PLHC-1 cell cultures was obtained with a submaximal dose of TCDD in combination with DEX. Furthermore, the authors observed a delay in obtaining a maximal degree of potentiation. Thus, a peak potentiation of CYP1A induction was observed after 48 h in cultures treated with 1 {micro}M BNF + 10 {micro}M DEX.

  17. A Multifaceted FISH Approach to Study Endogenous RNAs and DNAs within Native Nuclear and Cell Structure

    Science.gov (United States)

    Byron, Meg; Hall, Lisa L.; Lawrence, Jeanne B.

    2013-01-01

    Fluorescence in situ hybridization (FISH) is not a singular technique, but a battery of powerful and versatile tools to examine the distribution of endogenous genes and RNAs in precise context with each other and in relation to specific proteins or cell structures. This unit offers the details of highly sensitive and successful protocols that were initially developed largely in our lab and honed over a number of years. Our emphasis is on analysis of nuclear RNAs and DNA to address specific biological questions about nuclear structure, pre-mRNA metabolism or the role of non-coding RNAs, although cytoplasmic RNA detection is also provided and generally discussed. Multi-faceted molecular cytological approaches bring precise resolution and sensitive multi-color detection to illuminate the organization and functional roles of endogenous genes and their RNAs within the native structure of fixed cells. Solutions to several common technical pitfalls are discussed, as are cautions regarding the judicious use of digital imaging and the rigors of analyzing and interpreting complex molecular cytological results. PMID:23315927

  18. 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.)

  19. Towards the standardisation of the neuroblastoma (neuro-2a) cell-based assay for ciguatoxin-like toxicity detection in fish: application to fish caught in the Canary Islands.

    Science.gov (United States)

    Caillaud, A; Eixarch, H; de la Iglesia, P; Rodriguez, M; Dominguez, L; Andree, K B; Diogène, J

    2012-01-01

    The ouabain/veratridine-dependent neuroblastoma (neuro-2a) cell-based assay (CBA) was applied for the determination of the presence of ciguatoxin (CTX)-like compounds in ciguatera-suspected fish samples caught in the Canary Islands. In order to avoid matrix interferences the maximal concentration of wet weight fish tissue exposed to the neuro-2a cells was set at 20 mg tissue equivalent (TE) ml(-1) according to the sample preparation procedure applied. In the present study, the limit of quantification (LOQ) of CTX1B equivalents in fish extract was set at the limit of detection (LOD), being defined as the concentration of CTX1B equivalents inhibiting 20% cell viability (IC(20)). The LOQ was estimated as 0.0096 ng CTX1B eq.g TE(-1) with 23-31% variability between experiments. These values were deemed sufficient even though quantification given at the IC(50) (the concentration of CTX1B equivalents inhibiting 50% cell viability) is more accurate with a variability of 17-19% between experiments. Among the 13 fish samples tested, four fish samples were toxic to the neuro-2a cells with estimations of the content in CTX1B g(-1) of TE ranging from 0.058 (± 0.012) to 6.23 (± 0.713) ng CTX1B eq.g TE(-1). The high sensitivity and specificity of the assay for CTX1B confirmed its suitability as a screening tool of CTX-like compounds in fish extracts at levels that may cause ciguatera fish poisoning. Species identification of fish samples by DNA sequence analysis was conducted in order to confirm tentatively the identity of ciguatera risk species and it revealed some evidence of inadvertent misidentification. Results presented in this study are a contribution to the standardisation of the neuro-2a CBA and to the risk analysis for ciguatera in the Canary Islands.

  20. The impact of mitochondrial and thermal stress on the bioenergetics and reserve respiratory capacity of fish cell lines.

    Science.gov (United States)

    Beck, Benjamin H; Fuller, S Adam

    2012-12-01

    Various stressors affect the health of wild and cultured fish and can cause metabolic disturbances that first manifest at the cellular level. Here, we sought to further our understanding of cellular metabolism in fish by examining the metabolic responses of cell lines derived from channel catfish Ictalurus puntatus (CCO), white bass Morone chrysops (WBE), and fathead minnow Pimephales promelas (EPC) to both mitochondrial and thermal stressors. Using extracellular flux (EF) technology, we simultaneously measured the oxygen consumption rate (OCR; a measure of mitochondrial function) and extracellular acidification rate (ECAR; a surrogate of glycolysis) in each cell type. We performed a mitochondrial function protocol whereby compounds modulating different components of mitochondrial respiration were sequentially exposed to cells. This provided us with basal and maximal OCR, OCR linked to ATP production, OCR from ion movement across the mitochondrial inner membrane, the reserve capacity, and OCR independent of the electron transport chain. After heat shock, EPC and CCO significantly decreased OCR and all three cell lines modestly increased ECAR. After heat shock, the reserve capacity, the mitochondrial energetic reserve used to cope with stress and increased bioenergetic demand, was unaffected in EPC and CCO and completely abrogated in WBE. These findings provide proof-of-concept experimental data that further highlight the utility of fish cell lines as tools for modeling bioenergetics.

  1. 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...

  2. Cloning, Expression and Characterization of Zebra Fish Ferroportin in Hek 293T Cell Line

    Directory of Open Access Journals (Sweden)

    A Memarnejadian

    2012-01-01

    Full Text Available Background: Ferroportin (Fpn, a regulator of iron homeostasis is a conserved membrane protein that exports iron across the enterocytes, macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival of microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immunity and pathogenesis of micoorganisms.Methods: To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP-N1. The final resulted plasmid (pEGFP-ZFpn was used for expression of Fpn-EGFP protein in Hek 293T cells.Results: The expression was confirmed by appearance of fluorescence in Hek 293 T cells. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model, NetOGlyc 3.1 and NetNGlyc 3.1 servers. The obtained Fpn from indian zebrafish also contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 O-glycosylated amino acids.Conclusion: The recombinant Fpn from Indian zebra fish was successfully expressed in Hek 293 cell line. Although the discrepancy in two amino acids was observed in our produced Fpn and resulted in an additional O-glycosylation site, but had no effect on the topology of the protein compared to other Fpn described by other researchers. Therefore this construct can be used in future iron studies.

  3. 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)

  4. 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.

  5. 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...

  6. 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.

  7. Inhibition of Reporter Genes by Small Interfering RNAs in Cell Culture and Living Fish

    DEFF Research Database (Denmark)

    Larashati, Sekar; Schyth, Brian Dall; Lorenzen, Niels

    of reporter gene expression was found between individual fish but it seems that in glass catfish, siRNAs are able to reduce reporter gene expression in the muscle showing that it is possible to use siRNA as technology to target genes locally in living fish. In parallel experiments, which will not be reported...

  8. 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.)

  9. 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

  10. Inhibitory effect and cell damage on bacterial flora of fish caused by chitosan, nisin and sodium lactate.

    Science.gov (United States)

    Schelegueda, Laura Inés; Zalazar, Aldana Lourdes; Gliemmo, María Fernanda; Campos, Carmen Adriana

    2016-02-01

    The effect of the combined use of chitosan, nisin and sodium lactate on the growth of Listeria innocua, Shewanella putrefaciens and psychrophilic bacteria isolated from fish was investigated in broth by means of minimum inhibitory concentrations (MIC). Furthermore, the sites of cell-injury caused by mentioned antimicrobials and their combinations on L. innocua and S. putrefaciens were studied. MIC of antimicrobial mixtures were evaluated by Berembaum design and check board method. Antimicrobials' sites of injury were investigated by the evaluation of cell constituents' release, cell surface hydrophobicity and differential scanning calorimetry. Results depended on antimicrobial used; several combinations inhibited the growth of L. innocua and S. putrefaciens and all combinations inhibited psychrophilic bacteria. Besides, some mixtures showed synergistic effects. All the mixtures affected ribosomes and DNA of the studied bacteria. Regarding cellular envelope, antimicrobials acted according to the structural characteristics of target microorganisms. Cell damage was higher when antimicrobials were combined, which could explain the observed synergistic effects. This study demonstrates and justifies the synergistic action of chitosan, nisin and sodium lactate on the inhibition of microorganisms related to fish spoilage and remarks the promissory use of the synergic combination of antimicrobials for fish preservation.

  11. Elucidating fish oil-induced milk fat depression in dairy sheep: Milk somatic cell transcriptome analysis

    Science.gov (United States)

    Suárez-Vega, Aroa; Toral, Pablo G.; Gutiérrez-Gil, Beatriz; Hervás, Gonzalo; Arranz, Juan José; Frutos, Pilar

    2017-01-01

    In this study, RNA sequencing was used to obtain a comprehensive profile of the transcriptomic changes occurring in the mammary gland of lactating sheep suffering from fish oil-induced milk fat depression (FO-MFD). The milk somatic cell transcriptome analysis of four control and four FO-MFD ewes generated an average of 42 million paired-end reads per sample. In both conditions, less than 220 genes constitute approximately 89% of the total counts. These genes, which are considered as core genes, were mainly involved in cytoplasmic ribosomal proteins and electron transport chain pathways. In total, 117 genes were upregulated, and 96 genes were downregulated in FO-MFD samples. Functional analysis of the latter indicated a downregulation of genes involved in the SREBP signaling pathway (e.g., ACACA, ACSL, and ACSS) and Gene Ontology terms related to lipid metabolism and lipid biosynthetic processes. Integrated interpretation of upregulated genes indicated enrichment in genes encoding plasma membrane proteins and proteins regulating protein kinase activity. Overall, our results indicate that FO-MFD is associated with the downregulation of key genes involved in the mammary lipogenesis process. In addition, the results also suggest that this syndrome may be related to upregulation of other genes implicated in signal transduction and codification of transcription factors. PMID:28378756

  12. Proteomic analysis of blood cells in fish exposed to chemotherapeutics: evidence for long term effects.

    Science.gov (United States)

    Pierrard, Marie-Aline; Kestemont, Patrick; Phuong, Nguyen Thanh; Tran, Minh Phu; Delaive, Edouard; Thezenas, Marie-Laëtitia; Dieu, Marc; Raes, Martine; Silvestre, Frédéric

    2012-04-18

    Proteomics technology are increasingly used in ecotoxicological studies to characterize and monitor biomarkers of exposure. The present study aims at identifying long term effects of malachite green (MG) exposure on the proteome of peripheral blood mononuclear cells (PBMC) from the Asian catfish, Pangasianodon hypophthalmus. A common (0.1 ppm) concentration for therapeutic treatment was applied twice with a 72 h interval. PBMC were collected directly at the end of the second bath of MG (T1) and after 1 month of decontamination (T2). Analytical 2D-DIGE gels were run and a total of 2551±364 spots were matched. Among them, MG induced significant changes in abundance of 116 spots with no recovery after one month of decontamination. Using LC-MS/MS and considering single identification per spot, we could identify 25 different proteins. Additionally, MG residues were measured in muscle and in blood indicating that leuco-MG has almost totally disappeared after one month of decontamination. This work highlights long term effects of MG treatment on the PBMC proteome from fish intended for human consumption.

  13. Evolutionary and functional relationships of B cells from fish and mammals: insights into their novel roles in phagocytosis and presentation of particulate antigen.

    Science.gov (United States)

    Sunyer, J Oriol

    2012-06-01

    The evolutionary origins of Ig-producing B cells appear to be linked to the emergence of fish in this planet. There are three major classes of living fish species, which from most primitive to modern they are referred to as agnathan (e.g., lampreys), Chondrichthyes (e.g., sharks), and teleost fish (e.g., rainbow trout). Agnathans do not have immunoglobulin- producing B cells, however these fish contain a subset of lymphocytes-like cells producing type B variable lymphocyte receptors (VLRBs) that appear to act as functional analogs of immunoglobulins. Chondrichthyes fish represent the most primitive living species containing bona-fide immunoglobulin-producing B cells. Their B cells are known to secrete three types of antibodies, IgM, IgW and IgNAR. Teleost fish are also called bony fish since they represent the most ancient living species containing true bones. Teleost B cells produce three different immunoglobulin isotypes, IgM, IgD and the recently described IgT. While teleost IgM is the principal player in systemic immunity, IgT appears to be a teleost immunoglobulin class specialized in mucosal immune responses. Thus far, three major B cell lineages have been described in teleost, those expressing either IgT or IgD, and the most common lineage which co-expresses IgD and IgM. A few years ago, the study of teleost fish B cells revealed for the first time in vertebrates the existence of B cell subsets with phagocytic and intracellular bactericidal capacities. This finding represented a paradigm shift as professional phagocytosis was believed to be exclusively performed by some cells of the myeloid lineage (i.e., macrophages, monocytes, neutrophils). This phagocytic capacity was also found in amphibians and reptiles, suggesting that this innate capacity was evolutionarily conserved in certain B cell subsets of vertebrates. Recently, the existence of subsets of B cells with phagocytic and bactericidal abilities have also been confirmed in mammals. Moreover, it has

  14. Business plan Fish feed factory Tete Zambezi Valley, Mozambique

    NARCIS (Netherlands)

    Meer, van der Magnus; Boon, Hans; Brouwer, Herman

    2015-01-01

    This business plan has been prepared for local entrepreneurs who would like to expand their business portfolio or to start a value chain business in the aquaculture sector in the region of Tete, Mozambique.

  15. Ex vivo transfection of trout pronephros leukocytes, a model for cell culture screening of fish DNA vaccine candidates.

    Science.gov (United States)

    Ortega-Villaizan, M; Martinez-Lopez, A; Garcia-Valtanen, P; Chico, V; Perez, L; Coll, J M; Estepa, A

    2012-09-07

    DNA vaccination opened a new era in controlling and preventing viral diseases since DNA vaccines have shown to be very efficacious where some conventional vaccines have failed, as it occurs in the case of the vaccines against fish novirhabdoviruses. However, there is a big lack of in vitro model assays with immune-related cells for preliminary screening of in vivo DNA vaccine candidates. In an attempt to solve this problem, rainbow trout pronephros cells in early primary culture were transfected with two plasmid DNA constructions, one encoding the green fluorescent protein (GFP) and another encoding the viral haemorrhagic septicaemia virus (VHSV) glycoprotein G (G(VHSV)) - the only viral antigen which has conferred in vivo protection. After assessing the presence of GFP- and G(VHSV)-expressing cells, at transcription and protein levels, the immune response in transfected pronephros cells was evaluated. At 24h post-transfection, G(VHSV) up-regulated migm and tcr transcripts expression, suggesting activation of B and T cells, as well, a high up-regulation of tnfα gene was observed. Seventy-two hours post-transfection, we detected the up-regulation of mx and tnfα genes transcripts and Mx protein which correlated with the induction of an anti-VHSV state. All together we have gathered evidence for successful transfection of pronephros cells with pAE6G, which correlates with in vivo protection results, and is less time-consuming and more rapid than in vivo assays. Therefore, this outcome opens the possibility to use pronephros cells in early primary culture for preliminary screening fish DNA vaccines as well as to further investigate the function that these cells perform in fish immune response orchestration after DNA immunisation.

  16. 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.

  17. 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.

  18. Fish Allergy

    Science.gov (United States)

    ... Old Feeding Your 1- to 2-Year-Old Fish Allergy KidsHealth > For Parents > Fish Allergy Print A ... From Home en español Alergia al pescado About Fish Allergy A fish allergy is not exactly the ...

  19. 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

  20. Assessing proteinase K resistance of fish prion proteins in a scrapie-infected mouse neuroblastoma cell line.

    Science.gov (United States)

    Salta, Evgenia; Kanata, Eirini; Ouzounis, Christos A; Gilch, Sabine; Schätzl, Hermann; Sklaviadis, Theodoros

    2014-11-13

    The key event in prion pathogenesis is the structural conversion of the normal cellular protein, PrP(C), into an aberrant and partially proteinase K resistant isoform, PrP(Sc). Since the minimum requirement for a prion disease phenotype is the expression of endogenous PrP in the host, species carrying orthologue prion genes, such as fish, could in theory support prion pathogenesis. Our previous work has demonstrated the development of abnormal protein deposition in sea bream brain, following oral challenge of the fish with natural prion infectious material. In this study, we used a prion-infected mouse neuroblastoma cell line for the expression of three different mature fish PrP proteins and the evaluation of the resistance of the exogenously expressed proteins to proteinase K treatment (PK), as an indicator of a possible prion conversion. No evidence of resistance to PK was detected for any of the studied recombinant proteins. Although not indicative of an absolute inability of the fish PrPs to structurally convert to pathogenic isoforms, the absence of PK-resistance may be due to supramolecular and conformational differences between the mammalian and piscine PrPs.

  1. Feasibility of using fluorescence in situ hybridization (FISH) to detect early gene changes in sputum cells from uranium miners

    Energy Technology Data Exchange (ETDEWEB)

    Neft, R.E.; Rogers, J.L.; Belinsky, S.A. [and others

    1995-12-01

    Epidemiological studies have shown that combined exposure to radon progeny and tobacco smoke produce a greater than additive or synergistic increase in lung cancer risk. Lung cancer results from multiple genetic changes over a long period of time. An early change that occurs in lung cancer is trisomy 7 which is found in 50% of non-small cell lung cancer and in the far margins of resected lung tumors. The 80% mortality associated with lung cancer is in part related to the high proportion of patients who present with an advanced, unresectable tumor. Therefore, early detection of patients at risk for tumor development is critical to improve treatment of this disease. Currently, it is difficult to detect lung cancer early while it is still amendable by surgery. Saccomanno, G. has shown that premalignant cytologic changes in sputum cells collected from uranium miners can be detected by a skilled, highly trained cytopathologist. A more objective alternative for identifying premalignant cells in sputum may be to determine whether an early genetic change such as trisomy 7 is present in these cells. Fluorescence in situ hybridization (FISH) can be used to identify cells with trisomy 7. The results of this investigation indicate that FISH may prove to be an accurate, efficient method to test at-risk individuals for genetic alterations in bronchial epithelial cells from sputum.

  2. Studies of the Ecophysiology of Single Cells in Microbial Communities by (Quantitative) Microautoradiography and Fluorescence In Situ Hybridization (MAR-FISH)

    DEFF Research Database (Denmark)

    Nierychlo, Marta; Nielsen, Jeppe Lund; Nielsen, Per Halkjær

    2015-01-01

    Microautoradiography (MAR) in combination with fluorescence in situ hybridization (FISH) is a powerful method of obtaining information about the ecophysiology of probe-defined single cells in mixed microbial communities. The incorporation of radiolabelled substrates can be quantified by automated...

  3. 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.

  4. 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.

  5. Seasonal morphological and biochemical changes of Dahlgren cells implies a potential role of the caudal neurosecretory system (CNSS) in the reproduction cycle of teleostean fish.

    Science.gov (United States)

    Chen, Heng; Mu, Rui

    2008-03-01

    The purpose of this mini-review is to summarize recent research on the seasonal morphological and biochemical changes of Dahlgren cells in the caudal neurosecretory system (CNSS) of the freshwater teleosts carp Carassius auratus. The quantitative proof for these seasonal changes in the morphology and biochemistry of Dahlgren cells reflects the relationship between the CNSS and the reproduction cycle of fish and implies that the CNSS is probably involved in the reproduction process of fish.

  6. 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.

  7. Validation of interphase fluorescence in situ hybridization (iFISH for multiple myeloma using CD138 positive cells

    Directory of Open Access Journals (Sweden)

    Renata Kiyomi Kishimoto

    2016-06-01

    Full Text Available ABSTRACT BACKGROUND: Multiple myeloma is a plasma cell neoplasm with acquired genetic abnormalities of clinical and prognostic importance. Multiple myeloma differs from other hematologic malignancies due to a high fraction of low proliferating malignant plasma cells and the paucity of plasma cells in bone marrow aspiration samples, making cytogenetic analysis a challenge. An abnormal karyotype is found in only one-third of patients with multiple myeloma and interphase fluorescence in situ hybridization is the most useful test for studying the chromosomal abnormalities present in almost 90% of cases. However, it is necessary to study the genetic abnormalities in plasma cells after their identification or selection by morphology, immunophenotyping or sorting. Other challenges are the selection of the most informative FISH panel and determining cut-off levels for FISH probes. This study reports the validation of interphase fluorescence in situ hybridization using CD138 positive cells, according to proposed guidelines published by the European Myeloma Network (EMN in 2012. METHOD: Bone marrow samples from patients with multiple myeloma were used to standardize a panel of five probes [1q amplification, 13q14 deletion, 17p deletion, t(4;14, and t(14;16] in CD138+ cells purified by magnetic cell sorting. RESULTS: This test was validated with a low turnaround time and good reproducibility. Five of six samples showed genetic abnormalities. Monosomy/deletion 13 plus t(4;14 were found in two cases. CONCLUSION: This technique together with magnetic cell sorting is effective and can be used in the routine laboratory practice. In addition, magnetic cell sorting provides a pure plasma cell population that allows other molecular and genomic studies.

  8. Detection of denitrification genes by in situ rolling circle amplification - fluorescence in situ hybridization (in situ RCA-FISH) to link metabolic potential with identity inside bacterial cells

    DEFF Research Database (Denmark)

    Hoshino, Tatsuhiko; Schramm, Andreas

    2010-01-01

    A target-primed in situ rolling circle amplification (in situ RCA) protocol was developed for detection of single-copy genes inside bacterial cells and optimized with Pseudomonas stutzeri, targeting nitrite and nitrous oxide reductase genes (nirS and nosZ). Two padlock probes were designed per gene...... as Candidatus Accumulibacter phosphatis by combining in situ RCA-FISH with 16S rRNA-targeted FISH. While not suitable for quantification because of its low detection frequency, in situ RCA-FISH will allow to link metabolic potential with 16S rRNA (gene)-based identification of single microbial cells....

  9. Influence of cholesterol and fish oil dietary intake on nitric oxide-induced apoptosis in vascular smooth muscle cells.

    Science.gov (United States)

    Perales, Sonia; Alejandre, Ma José; Palomino-Morales, Rogelio; Torres, Carolina; Linares, Ana

    2010-04-01

    Apoptosis of vascular smooth muscle cells (SMC) is critically involved in the progression of atherosclerosis. We previously reported that dietary cholesterol intake induces changes in SMC at molecular and gene expression levels. The objectives of the present study were to investigate the differential response to nitric oxide of vascular SMC obtained from chicks after cholesterol and fish oil dietary intake and to examine effects on the main pro-apoptotic and anti-apoptotic genes. Dietary cholesterol intake reduced the Bcl-2/Bax (anti-apoptotic/pro-apoptotic) protein ratio in SMC, making them more susceptible to apoptosis. When cholesterol was withdrawn and replaced with a fish oil-enriched diet, the Bcl-xl/Bax protein ratio significantly increased, reversing the changes induced by cholesterol. The decrease in c-myc gene expression after apoptotic stimuli and the increase in Bcl-xl/Bax ratio indicate that fish oil has a protective role against apoptosis in SMC. Nitroprussiate-like nitric oxide donors exerted an intensive action on vascular SMC cultures. However, SMC-C (isolated from animals fed with control diet) and SMC-Ch (isolated from animals fed with cholesterol-enriched diet) responded differently to nitric oxide, especially in their bcl-2 and bcl-xl gene expression. SMC isolated from animals fed with cholesterol-enriched and then fish oil-enriched diet (SMC-Ch-FO cultures) showed an intermediate apoptosis level (Bcl-2/Bax ratio) between SMC-C and SMC-Ch, induction of c-myc expression and elevated p53 expression. These findings indicate that fish oil protects SMC against apoptosis.

  10. Development of iPS (induced pluripotent stem cells) using natural product from extract of fish oocyte to provide stem cell for regenerative therapy

    Science.gov (United States)

    Meilany, Sofy; Firdausiyah, Qonitha S.; Naroeni, Aroem

    2017-02-01

    In this study, we developed a method to induce pluripotency of adult cells (fibroblast) into stem cells using a natural product, extract of fish oocyte, by comparing the extract concentration, 1 mg/ml and 2 mg/ml. The analyses were done by measuring the Nanog gene expression in cells using qPCR and detecting fibroblast marker anti H2-KK. The results revealed existence of a colony of stem cells in the cell that was induced with 2mg/ml concentration of oocytes. Nanoggene expression was analyzed by qPCR and the results showed expression of Nanog gene compared to the control. Analysis of result of fibroblast using Tali Cytometer and anti H2KK antibody showed loss of expression of Anti H2KK meaning there was transformation from fibroblast type cell to pluripotent cell type.

  11. Genotoxicity, potential cytotoxicity and cell uptake of titanium dioxide nanoparticles in the marine fish Trachinotus carolinus (Linnaeus, 1766)

    Energy Technology Data Exchange (ETDEWEB)

    Vignardi, Caroline P., E-mail: carolpatvig@usp.br [Department of Biological Oceanography, Oceanographic Institute, University of São Paulo, Praça do Oceanogáfico 191, Cidade Universitária, Butantã, São Paulo, SP 05508900 (Brazil); Hasue, Fabio M., E-mail: humbigutis@gmail.com [Department of Biological Oceanography, Oceanographic Institute, University of São Paulo, Praça do Oceanogáfico 191, Cidade Universitária, Butantã, São Paulo, SP 05508900 (Brazil); Sartório, Priscila V., E-mail: pri.sartorio@gmail.com [Department of Biological Oceanography, Oceanographic Institute, University of São Paulo, Praça do Oceanogáfico 191, Cidade Universitária, Butantã, São Paulo, SP 05508900 (Brazil); Cardoso, Caroline M., E-mail: camargonato@gmail.com [Department of Biological Oceanography, Oceanographic Institute, University of São Paulo, Praça do Oceanogáfico 191, Cidade Universitária, Butantã, São Paulo, SP 05508900 (Brazil); Machado, Alex S.D., E-mail: mamiferomarinho@gmail.com [Faculty of Veterinary Medicine, Integrated College North of Minas Osmane Barbosa Avenue, 11111, JK, Montes Claros, MG 39404006 (Brazil); and others

    2015-01-15

    Highlights: • TiO{sub 2}–NP cytogenotoxicity and cell uptake in marine fish was studied. • TiO{sub 2}–NP suspension was in primary particle, agglomerated and aggregated form. • TiO{sub 2}–NP genotoxicity was time/dose dependent and may induce cell uptake. • Methodology proved to be efficient for evaluating the toxic effect of TiO{sub 2}–NP. - Abstract: Nanoparticles have physicochemical characteristics that make them useful in areas such as science, technology, medicine and in products of everyday use. Recently the manufacture and variety of these products has grown rapidly, raising concerns about their impact on human health and the environment. Adverse effects of exposure to nanoparticles have been reported for both terrestrial and aquatic organisms, but the toxic effects of the substances on marine organisms remain poorly understood. The main aim of this study was to evaluate the genotoxicity of TiO{sub 2}–NP in the marine fish Trachinotus carolinus, through cytogenotoxic methods. The fish received two different doses of 1.5 μg and 3.0 μg–TiO{sub 2}–NP g{sup −1} by intraperitoneal injection. Blood samples were collected to analyze erythrocyte viability using the Trypan Blue exclusion test, comet assay (pH > 13), micronucleus (MN) and other erythrocyte nuclear abnormalities (ENA) 24, 48 and 72 h after injection. The possible cell uptake of TiO{sub 2}–NP in fish injected with the higher dose was investigated after 72 h using transmission electron microscopy (TEM). The results showed that TiO{sub 2}–NP is genotoxic and potentially cytotoxic for this species, causing DNA damage, inducing the formation of MN and other ENA, and decreasing erythrocyte viability. TEM examination revealed that cell uptake of TiO{sub 2}–NP was mainly in the kidney, liver, gills and to a lesser degree in muscle. To the extent of the authors’ knowledge, this is the first in vivo study of genotoxicity and other effects of TiO{sub 2}–NP in a marine fish.

  12. Randomized controlled trial examining the effects of fish oil and multivitamin supplementation on the incorporation of n-3 and n-6 fatty acids into red blood cells.

    Science.gov (United States)

    Pipingas, Andrew; Cockerell, Robyn; Grima, Natalie; Sinclair, Andrew; Stough, Con; Scholey, Andrew; Myers, Stephen; Croft, Kevin; Sali, Avni; Pase, Matthew P

    2014-05-14

    The present randomized, placebo-controlled, double-blind, parallel-groups clinical trial examined the effects of fish oil and multivitamin supplementation on the incorporation of n-3 and n-6 fatty acids into red blood cells. Healthy adult humans (n = 160) were randomized to receive 6 g of fish oil, 6 g of fish oil plus a multivitamin, 3 g of fish oil plus a multivitamin or a placebo daily for 16 weeks. Treatment with 6 g of fish oil, with or without a daily multivitamin, led to higher eicosapentaenoic acid (EPA) composition at endpoint. Docosahexaenoic acid (DHA) composition was unchanged following treatment. The long chain LC n-3 PUFA index was only higher, compared to placebo, in the group receiving the combination of 6 g of fish oil and the multivitamin. Analysis by gender revealed that all treatments increased EPA incorporation in females while, in males, EPA was only significantly increased by the 6 g fish oil multivitamin combination. There was considerable individual variability in the red blood cell incorporation of EPA and DHA at endpoint. Gender contributed to a large proportion of this variability with females generally showing higher LC n-3 PUFA composition at endpoint. In conclusion, the incorporation of LC n-3 PUFA into red blood cells was influenced by dosage, the concurrent intake of vitamin/minerals and gender.

  13. Randomized Controlled Trial Examining the Effects of Fish Oil and Multivitamin Supplementation on the Incorporation of n-3 and n-6 Fatty Acids into Red Blood Cells

    Directory of Open Access Journals (Sweden)

    Andrew Pipingas

    2014-05-01

    Full Text Available The present randomized, placebo-controlled, double-blind, parallel-groups clinical trial examined the effects of fish oil and multivitamin supplementation on the incorporation of n-3 and n-6 fatty acids into red blood cells. Healthy adult humans (n = 160 were randomized to receive 6 g of fish oil, 6 g of fish oil plus a multivitamin, 3 g of fish oil plus a multivitamin or a placebo daily for 16 weeks. Treatment with 6 g of fish oil, with or without a daily multivitamin, led to higher eicosapentaenoic acid (EPA composition at endpoint. Docosahexaenoic acid (DHA composition was unchanged following treatment. The long chain LC n-3 PUFA index was only higher, compared to placebo, in the group receiving the combination of 6 g of fish oil and the multivitamin. Analysis by gender revealed that all treatments increased EPA incorporation in females while, in males, EPA was only significantly increased by the 6 g fish oil multivitamin combination. There was considerable individual variability in the red blood cell incorporation of EPA and DHA at endpoint. Gender contributed to a large proportion of this variability with females generally showing higher LC n-3 PUFA composition at endpoint. In conclusion, the incorporation of LC n-3 PUFA into red blood cells was influenced by dosage, the concurrent intake of vitamin/minerals and gender.

  14. Ecotoxicological evaluation of sodium fluoroacetate on aquatic organisms and investigation of the effects on two fish cell lines.

    Science.gov (United States)

    Zurita, Jorge L; Jos, Angeles; Cameán, Ana M; Salguero, Manuel; López-Artíguez, Miguel; Repetto, Guillermo

    2007-02-01

    Sodium monofluoroacetate (compound 1080) is one of the most potent pesticides. It is also a metabolite of many other fluorinated compounds, including anticancer agents, narcotic analgesics, pesticides or industrial chemicals. Other sources of water contamination are the atmospheric degradation of hydrofluorocarbons and hydrochlorofluorocarbons. However, there is little information available about the adverse effects of sodium fluoroacetate in aquatic organisms. Firstly, the bacterium Vibrio fischeri (decomposer), the alga Chlorella vulgaris (1st producer) and the cladoceran Daphnia magna (1st consumer) were used for the ecotoxicological evaluation of SMFA. The most sensitive models were C. vulgaris and D. magna, with a NOAEL of 0.1 and an EC50 of 0.5 mM at 72 h, respectively. According to the results after the acute exposure and due to its high biodegradation rate and low bioaccumulation potential, sodium fluoroacetate is most unlikely to produce deleterious effects to aquatic organisms. Secondly, two fish cell lines were employed to investigate the effects and mechanisms of toxicity in tissues from 2nd consumers. The hepatoma fish cell line PLHC-1 was more sensitive to SMFA than the fibroblast-like fish cell line RTG-2, being the uptake of neutral red the most sensitive bioindicator. Lysosomal function, succinate dehydrogenase and acetylcholinesterase activities were inhibited, glucose-6-phosphate dehydrogenase activity was particularly stimulated, and metallothionein and ethoxyresorufin-O-deethylase levels were not modified. Intense hydropic degeneration, macrovesicular steatosis and death mainly by necrosis but also by apoptosis were observed. Moreover, sulphydryl groups and oxidative stress could be involved in PLHC-1 cell death induced by SMFA more than changes in calcium homeostasis.

  15. Yeast as factory and factotum.

    Science.gov (United States)

    Dixon, B

    2000-02-01

    After centuries of vigorous activity in making fine wines, beers and breads, Saccharomyces cerevisiae is now acquiring a rich new portfolio of skills, bestowed by genetic manipulation. As shown in a recent shop-window of research supported by the European Commission, yeasts will soon be benefiting industries as diverse as fish farming, pharmaceuticals and laundering.

  16. 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

  17. Meat and fish consumption and the risk of renal cell carcinoma in the European prospective investigation into cancer and nutrition.

    Science.gov (United States)

    Rohrmann, Sabine; Linseisen, Jakob; Overvad, Kim; Lund Würtz, Anne Mette; Roswall, Nina; Tjonneland, Anne; Boutron-Ruault, Marie-Christine; Racine, Antoine; Bastide, Nadia; Palli, Domenico; Agnoli, Claudia; Panico, Salvatore; Tumino, Rosario; Sacerdote, Carlotta; Weikert, Steffen; Steffen, Annika; Kühn, Tilman; Li, Kuanrong; Khaw, Kay-Tee; Wareham, Nicholas J; Bradbury, Kathryn E; Peppa, Eleni; Trichopoulou, Antonia; Trichopoulos, Dimitrios; Bueno-de-Mesquita, H Bas; Peeters, Petra H M; Hjartåker, Anette; Skeie, Guri; Weiderpass, Elisabete; Jakszyn, Paula; Dorronsoro, Miren; Barricarte, Aurelio; Santiuste de Pablos, Carmen; Molina-Montes, Esther; de la Torre, Ramón Alonso; Ericson, Ulrika; Sonestedt, Emily; Johansson, Mattias; Ljungberg, Börje; Freisling, Heinz; Romieu, Isabelle; Cross, Amanda J; Vergnaud, Anne-Claire; Riboli, Elio; Boeing, Heiner

    2015-03-01

    Renal cell cancer (RCC) incidence varies worldwide with a higher incidence in developed countries and lifestyle is likely to contribute to the development of this disease. We examined whether meat and fish consumption were related to the risk of RCC in the European Prospective Investigation into Cancer and Nutrition (EPIC). The analysis included 493,179 EPIC participants, recruited between 1992 and 2000. Until December 2008, 691 RCC cases have been identified. Meat and fish consumption was assessed at baseline using country-specific dietary assessment instruments; 24-hour recalls were applied in an 8% subsample for calibration purposes. Cox proportional hazards regression was used to calculate multivariable-adjusted hazard ratios (HR) and 95% confidence intervals (CI). Women with a high consumption of red meat (HR = 1.36, 95% CI 1.14-1.62; calibrated, per 50 g/day) and processed meat (HR = 1.78, 95% CI 1.05-3.03; calibrated, per 50 g/day) had a higher risk of RCC, while no association existed in men. For processed meat, the association with RCC incidence was prominent in premenopausal women and was lacking in postmenopausal women (p interaction = 0.02). Neither poultry nor fish consumption were statistically significantly associated with the risk of RCC. The results show a distinct association of red and processed meat consumption with incident RCC in women but not in men. A biological explanation for these findings remains unclear.

  18. 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.

  19. Stiffness changes of the cupula associated with the mechanics of hair cells in the fish lateral line.

    Science.gov (United States)

    van Netten, S M; Khanna, S M

    1994-02-15

    Cupular vibration in the lateral-line canal of fish was measured in response to motion of the fluid in the canal by laser-heterodyne interferometry. The results show that the mechanical output/input ratio of the cupula depends on the stimulus amplitude; the cupula thus behaves nonlinearly. The nonlinearity is due to the hair bundles, since it disappears when the cupula is uncoupled from the underlying hair cells. A model of cupular dynamics in which the behavior of the gating springs of the hair cells is incorporated predicts nonlinear responses that are similar to the measurements, suggesting that the nonlinear behavior of the cupula may be attributed to the opening and closing of the transduction channels of the hair cells.

  20. In vitro cytotoxicity assessment of imidazolium ionic liquids: biological effects in fish Channel Catfish Ovary (CCO) cell line.

    Science.gov (United States)

    Radošević, Kristina; Cvjetko, Marina; Kopjar, Nevenka; Novak, Rudjer; Dumić, Jerka; Srček, Višnja Gaurina

    2013-06-01

    Increasing interest in the application of ionic liquids as green replacement for volatile organic solvents emphasized the need for the evaluation of their toxic effects at different biological systems in order to reduce the risk for human health and environment. To our knowledge, effects of imidazolium ionic liquids on cellular level of fish cell lines have not been studied yet. The cytotoxicity of imidazolium ionic liquids containing different anions and alkyl chain lengths as the substituent at the cation ring towards the fish CCO cell line was determined by WST-1 proliferation assay. Morphological alterations were examined by fluorescent microscopy using acridine orange/ethidium bromide staining and flow cytometry analysis was also performed. The results showed concentration-dependent cytotoxicity of ionic liquids in CCO cells, related to the type of anion and alkyl chain length, while EC50 values showed moderate to high cytotoxicity of tested imidazolium ionic liquids. Distinct morphological changes observed under fluorescence microscope and data obtained by flow cytometry suggest that the toxicity of imidazolium ionic liquids with longer alkyl chains could be related to necrosis. Results presented in here may be helpful for filling existing gaps of knowledge about ionic liquids toxicity and their impact on aquatic environment.

  1. Fish Allergy

    Science.gov (United States)

    ... in a clear and consistent manner, so that consumers with food allergies and their caregivers can be informed as ... the menu, cross-contact with fish is possible. Ethnic ... fish. Avoid foods like fish sticks and anchovies. Some individuals with ...

  2. Fish Hearing.

    Science.gov (United States)

    Blaxter, J. H. S.

    1980-01-01

    Provides related information about hearing in fish, including the sensory stimulus of sound in the underwater environment, mechanoreceptors in fish, pressure perception and the swimbladder, specializations in sound conduction peculiar to certain fish families. Includes numerous figures. (CS)

  3. 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...

  4. The effects of fish oil and high or low linoleic acid intake on fatty acid composition of human peripheral blood mononuclear cells

    DEFF Research Database (Denmark)

    Damsgaard, C.T.; Frøkiær, Hanne; Lauritzen, L.

    2008-01-01

    -supplementation on tissue fatty acid composition. Healthy young men (it 64) were randomized to capsules with FO or olive oil (control) (4-4 (2-0-5-6) ml/d) and to either sunflower oil and margarine (S/B) or rapeseed oil and a butter spread (R/K) to provide a high or a low 18: 2n-6 intake. Diet was measured by 4-d weighed......Dietary intake of 18: 2n-6 and 18: 3n-3 may affect endogenous production and incorporation of n-3 long-chain PUFA (LCPUFA) from fish oils (170). This double-blinded controlled 2 x 2-factorial 8-week intervention investigates the effects of high and low 18: 2n-6 intake in combination with FO...

  5. The effects of fish oil and high or low linoleic acid intake on fatty acid composition of human peripheral blood mononuclear cells

    DEFF Research Database (Denmark)

    Damsgaard, Camilla Trab; Frøkiær, Hanne; Lauritzen, Lotte

    2008-01-01

    -supplementation on tissue fatty acid composition. Healthy young men (n 64) were randomized to capsules with FO or olive oil (control) (44 (20-56) ml/d) and to either sunflower oil and margarine (S/B) or rapeseed oil and a butter spread (R/K) to provide a high or a low 18: 2n-6 intake. Diet was measured by 4-d weighed......Dietary intake of 18: 2n-6 and 18: 3n-3 may affect endogenous production and incorporation of n-3 long-chain PUFA (LCPUFA) from fish oils (FO). This double-blinded controlled 2 £ 2-factorial 8-week intervention investigates the effects of high and low 18: 2n-6 intake in combination with FO...

  6. Graphene nanoplatelets spontaneously translocate into the cytosol and physically interact with cellular organelles in the fish cell line PLHC-1

    Energy Technology Data Exchange (ETDEWEB)

    Lammel, Tobias; Navas, José M., E-mail: jmnavas@inia.es

    2014-05-01

    Highlights: • We assessed the cytotoxicity and uptake of graphene nanomaterials in PLHC-1 cells. • GO and CXYG nanoplatelets caused physical injury of the plasma membrane. • GO and CXYG accumulated in the cytosol and interacted with cellular organelles. • PLHC-1 cells exposed to GO/CXYG demonstrated high ROS levels but low cytotoxicity. • ROS formation was related with GO/CXYG-induced structural damage of mitochondria. - Abstract: Graphene and graphene derivatives constitute a novel class of carbon-based nanomaterials being increasingly produced and used in technical and consumer applications. Release of graphene nanoplatelets during the life cycle of these applications may result in human and environmental exposure calling for assessment of their potential to cause harm to humans and wildlife. This study aimed to assess the toxicity of graphene oxide (GO) and carboxyl graphene (CXYG) nanoplatelets to non-mammalian species using the fish cell line PLHC-1 as in vitro model. The cytotoxicity of GO and CXYG was assessed using different assays measuring alterations in plasma membrane integrity, metabolic activity, and lysosomal and mitochondrial function. The induction of oxidative stress was assessed by measuring intracellular reactive oxygen species (ROS) levels. Interaction with the plasma membrane and internalization of nanoplatelets were investigated by electron microscopy. Graphene nanoplatelets spontaneously penetrated through the plasma membrane and accumulated in the cytosol, where they further interacted with mitochondrial and nuclear membranes. PLHC-1 cells demonstrated significantly reduced mitochondrial membrane potential (MMP) and increased ROS levels at 16 μg/ml GO and CXYG (72 h), but barely any decrease in cell viability. The observation of intracellular graphene accumulations not enclosed by membranes suggests that GO and CXYG internalization in fish hepatoma cells occurs through an endocytosis-independent mechanism.

  7. 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

  8. 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.

  9. A FISH-based method for assessment of HER-2 amplification status in breast cancer circulating tumor cells following CellSearch isolation

    Directory of Open Access Journals (Sweden)

    Frithiof H

    2016-11-01

    Full Text Available Henrik Frithiof,1 Kristina Aaltonen,1 Lisa Rydén2,3 1Division of Oncology and Pathology, 2Division of Surgery, Department of Clinical Sciences Lund, Lund University, Lund, 3Department of Surgery, Skåne University Hospital, Malmö, Sweden Introduction: Amplification of the HER-2/neu (HER-2 proto-oncogene occurs in 10%–15% of primary breast cancer, leading to an activated HER-2 receptor, augmenting growth of cancer cells. Tumor classification is determined in primary tumor tissue and metastatic biopsies. However, malignant cells tend to alter their phenotype during disease progression. Circulating tumor cell (CTC analysis may serve as an alternative to repeated biopsies. The Food and Drug Administration-approved CellSearch system allows determination of the HER-2 protein, but not of the HER-2 gene. The aim of this study was to optimize a fluorescence in situ hybridization (FISH-based method to quantitatively determine HER-2 amplification in breast cancer CTCs following CellSearch-based isolation and verify the method in patient samples. Methods: Using healthy donor blood spiked with human epidermal growth factor receptor 2 (HER-2-positive breast cancer cell lines, SKBr-3 and BT-474, and a corresponding negative control (the HER-2-negative MCF-7 cell line, an in vitro CTC model system was designed. Following isolation in the CellSearch system, CTC samples were further enriched and fixed on microscope slides. Immunocytochemical staining with cytokeratin and 4',6-diamidino-2'-phenylindole dihydrochloride identified CTCs under a fluorescence microscope. A FISH-based procedure was optimized by applying the HER2 IQFISH pharmDx assay for assessment of HER-2 amplification status in breast cancer CTCs. Results: A method for defining the presence of HER-2 amplification in single breast cancer CTCs after CellSearch isolation was established using cell lines as positive and negative controls. The method was validated in blood from breast cancer patients

  10. The effects of dietary fish oil on cell populations, cytokines, chemokines and chemokine receptors in healthy mice and mice with endotoxin-induced peritonitis

    OpenAIRE

    Hildur Hrönn Arnardóttir 1981

    2011-01-01

    Fish oil, rich in n-3 polyunsaturated fatty acids, has immunomodulatory properties and may have beneficial effects in several immune related disorders, including sepsis. Chemokines and chemokine receptors play a key role in the recruitment of specific populations of immune cells to the sites of infection or inflammation. The results from the Ph.D. project show that dietary fish oil decreased the proportion of classical monocytes(expressing the chemokine receptor CCR2) in blood from healthy mi...

  11. Poised transcription factories prime silent uPA gene prior to activation.

    Directory of Open Access Journals (Sweden)

    Carmelo Ferrai

    2010-01-01

    Full Text Available The position of genes in the interphase nucleus and their association with functional landmarks correlate with active and/or silent states of expression. Gene activation can induce chromatin looping from chromosome territories (CTs and is thought to require de novo association with transcription factories. We identify two types of factory: "poised transcription factories," containing RNA polymerase II phosphorylated on Ser5, but not Ser2, residues, which differ from "active factories" associated with phosphorylation on both residues. Using the urokinase-type plasminogen activator (uPA gene as a model system, we find that this inducible gene is predominantly associated with poised (S5p(+S2p(- factories prior to activation and localized at the CT interior. Shortly after induction, the uPA locus is found associated with active (S5p(+S2p(+ factories and loops out from its CT. However, the levels of gene association with poised or active transcription factories, before and after activation, are independent of locus positioning relative to its CT. RNA-FISH analyses show that, after activation, the uPA gene is transcribed with the same frequency at each CT position. Unexpectedly, prior to activation, the uPA loci internal to the CT are seldom transcriptionally active, while the smaller number of uPA loci found outside their CT are transcribed as frequently as after induction. The association of inducible genes with poised transcription factories prior to activation is likely to contribute to the rapid and robust induction of gene expression in response to external stimuli, whereas gene positioning at the CT interior may be important to reinforce silencing mechanisms prior to induction.

  12. Fish Immunoglobulins

    Science.gov (United States)

    Mashoof, Sara; Criscitiello, Michael F.

    2016-01-01

    The B cell receptor and secreted antibody are at the nexus of humoral adaptive immunity. In this review, we summarize what is known of the immunoglobulin genes of jawed cartilaginous and bony fishes. We focus on what has been learned from genomic or cDNA sequence data, but where appropriate draw upon protein, immunization, affinity and structural studies. Work from major aquatic model organisms and less studied comparative species are both included to define what is the rule for an immunoglobulin isotype or taxonomic group and what exemplifies an exception. PMID:27879632

  13. Several synthetic progestins disrupt the glial cell specific-brain aromatase expression in developing zebra fish

    OpenAIRE

    Cano-Nicolau, Joel; Garoche, Clementine; Hinfray, Nathalie; Pellegrini, Elisabeth; Boujrad, Noureddine; Pakdel, Farzad; Kah, Olivier; Brion, François

    2016-01-01

    International audience; The effects of some progestins on fish reproduction have been recently reported revealing the hazard of this class of steroidal pharmaceuticals. However, their effects at the central nervous system level have been poorly studied until now. Notwithstanding, progesterone, although still widely considered primarily a sex hormone, is an important agent affecting many central nervous system functions. Herein, we investigated the effects of a large set of synthetic ligands o...

  14. Amiloride causes changes in the mechanical properties of hair cell bundles in the fish lateral line similar to those induced by dihydrostreptomycin

    NARCIS (Netherlands)

    Wiersinga-Post, JEC; van Netten, SM

    1998-01-01

    Amiloride is a known blocker of the mechano-electrical transduction current in sensory hair cells. Measurements of cupular motion in the lateral line organ of fish now show that amiloride concurrently changes the micromechanical properties of the hair cell bundles. The effects of amiloride on the me

  15. 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...

  16. 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.

  17. Development of an Efficient Genome Editing Method by CRISPR/Cas9 in a Fish Cell Line.

    Science.gov (United States)

    Dehler, Carola E; Boudinot, Pierre; Martin, Samuel A M; Collet, Bertrand

    2016-08-01

    CRISPR/Cas9 system has been used widely in animals and plants to direct mutagenesis. To date, no such method exists for fish somatic cell lines. We describe an efficient procedure for genome editing in the Chinook salmon Oncorhynchus tshawytscha CHSE. This cell line was genetically modified to firstly overexpress a monomeric form of EGFP (cell line CHSE-E Geneticin resistant) and additionally to overexpress nCas9n, a nuclear version of Cas9 (cell line CHSE-EC, Hygromycin and Geneticin resistant). A pre-validated sgRNA was produced in vitro and used to transfect CHSE-EC cells. The EGFP gene was disrupted in 34.6 % of cells, as estimated by FACS and microscopy. The targeted locus was characterised by PCR amplification, cloning and sequencing of PCR products; inactivation of the EGFP gene by deletions in the expected site was validated in 25 % of clones. This method opens perspectives for functional genomic studies compatible with high-throughput screening.

  18. 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...

  19. Effect of Marine Omega 3 Fatty Acids on Methylmercury-Induced Toxicity in Fish and Mammalian Cells In Vitro

    Directory of Open Access Journals (Sweden)

    O. J. Nøstbakken

    2012-01-01

    Full Text Available Methylmercury (MeHg is a ubiquitous environmental contaminant which bioaccumulates in marine biota. Fish constitute an important part of a balanced human diet contributing with health beneficial nutrients but may also contain contaminants such as MeHg. Interactions between the marine n-3 fatty acids eicosapentaenoic acid (20:5n-3, EPA and docosahexaenoic acid (22:6n-3, DHA with MeHg-induced toxicity were investigated. Different toxic and metabolic responses were studied in Atlantic salmon kidney (ASK cell line and the mammalian kidney-derived HEK293 cell line. Both cell lines were preincubated with DHA or EPA prior to MeHg-exposure, and cell toxicity was assessed differently in the cell lines by MeHg-uptake in cells (ASK and HEK293, proliferation (HEK293 and ASK, apoptosis (ASK, oxidation of the red-ox probe roGFP (HEK293, and regulation of selected toxicological and metabolic transcriptional markers (ASK. DHA was observed to decrease the uptake of MeHg in HEK293, but not in ASK cells. DHA also increased, while EPA decreased, MeHg-induced apoptosis in ASK. MeHg exposure induced changes in selected metabolic and known MeHg biomarkers in ASK cells. Both DHA and MeHg, but not EPA, oxidized roGFP in HEK293 cells. In conclusion, marine n-3 fatty acids may ameliorate MeHg toxicity, either by decreasing apoptosis (EPA or by reducing MeHg uptake (DHA. However, DHA can also augment MeHg toxicity by increasing oxidative stress and apoptosis when combined with MeHg.

  20. In vivo regulation of colonic cell proliferation, differentiation, apoptosis, and P27Kip1 by dietary fish oil and butyrate in rats.

    Science.gov (United States)

    Hong, Mee Young; Turner, Nancy D; Murphy, Mary E; Carroll, Raymond J; Chapkin, Robert S; Lupton, Joanne R

    2015-11-01

    We have shown that dietary fish oil is protective against experimentally induced colon cancer, and the protective effect is enhanced by coadministration of pectin. However, the underlying mechanisms have not been fully elucidated. We hypothesized that fish oil with butyrate, a pectin fermentation product, protects against colon cancer initiation by decreasing cell proliferation and increasing differentiation and apoptosis through a p27(Kip1)-mediated mechanism. Rats were provided diets of corn or fish oil, with/without butyrate, and terminated 12, 24, or 48 hours after azoxymethane (AOM) injection. Proliferation (Ki-67), differentiation (Dolichos Biflorus Agglutinin), apoptosis (TUNEL), and p27(Kip1) (cell-cycle mediator) were measured in the same cell within crypts in order to examine the coordination of cell cycle as a function of diet. DNA damage (N(7)-methylguanine) was determined by quantitative IHC analysis. Dietary fish oil decreased DNA damage by 19% (P = 0.001) and proliferation by 50% (P = 0.003) and increased differentiation by 56% (P = 0.039) compared with corn oil. When combined with butyrate, fish oil enhanced apoptosis 24 hours after AOM injection compared with a corn oil/butyrate diet (P = 0.039). There was an inverse relationship between crypt height and apoptosis in the fish oil/butyrate group (r = -0.53, P = 0.040). The corn oil/butyrate group showed a positive correlation between p27(Kip1) expression and proliferation (r = 0.61, P = 0.035). These results indicate the in vivo effect of butyrate on apoptosis and proliferation is dependent on dietary lipid source. These results demonstrate the presence of an early coordinated colonocyte response by which fish oil and butyrate protects against colon tumorigenesis.

  1. Comparative in vitro study of cholinium-based ionic liquids and deep eutectic solvents toward fish cell line.

    Science.gov (United States)

    Radošević, Kristina; Železnjak, Jelena; Cvjetko Bubalo, Marina; Radojčić Redovniković, Ivana; Slivac, Igor; Gaurina Srček, Višnja

    2016-09-01

    With the advent of ionic liquids, much was expected concerning their applicability as an alternative to organic solvents in the chemical technology and biotechnology fields. However, the most studied and commonly used ionic liquids based on imidazolium and pyridinium were found not to be as environmentally friendly as it was first expected. Therefore, a new generation of alternative solvents named natural ionic liquids and deep eutectic solvents, composed of natural and/or renewable compounds, have come into focus in recent years. Since the number of newly synthesized chemicals increases yearly, simple and reliable methods for their ecotoxicological assessment are necessary. Permanent fish cell lines can serve as a test system for the evaluation of a chemical's cytotoxicity. This paper presents research results on the cytotoxic effects on Channel Catfish Ovary (CCO) cell line induced by fifteen cholinium-based ionic liquids and deep eutectic solvents. Based on the decrease in cell viability, the most obvious toxic effect on CCO cells was caused by ionic liquid choline oxalate, while other solvents tested exhibited low cytotoxicity. Therefore, we can conclude that cholinium-based ionic liquids and deep eutectic solvents are comparatively less toxic to CCO cells than conventional ionic liquids.

  2. Copper toxicity in gills of the teleost fish, Oreochromis niloticus: Effects in apoptosis induction and cell proliferation

    Energy Technology Data Exchange (ETDEWEB)

    Monteiro, Sandra Mariza, E-mail: smonteir@utad.pt [Department of Biology and Environment-CITAB, University of Tras-os-Montes and Alto Douro, Apartado 1013, 5001-801 Vila Real (Portugal); Santos, Nuno M.S. dos [Institute for Molecular and Cell Biology, University of Porto (Portugal); Calejo, Margarida [Lab Cell Biology - ICBAS, University of Porto (Portugal); Fontainhas-Fernandes, Antonio [Department of Biology and Environment-CITAB, University of Tras-os-Montes and Alto Douro, Apartado 1013, 5001-801 Vila Real (Portugal); Sousa, Mario [Lab Cell Biology - ICBAS, University of Porto (Portugal); Centre for Reproductive Genetics Alberto Barros, Porto (Portugal)

    2009-09-14

    Recent in vitro studies have demonstrated that copper may induce apoptosis triggering the activation of caspase-3, a central effector of apoptotic cell death. However, the precise mechanism of copper-induced apoptosis is still unclear, even less so in Oreochromis niloticus where no caspase genes have been reported so far. This study aimed to assess the in vivo role of copper in apoptosis induction on O. niloticus gill, simultaneously contributing to elucidate the mechanism of copper-induced apoptosis. Caspase-3 gene was partially sequenced and, after in vivo exposures to 40 and 400 {mu}g L{sup -1} of copper, its mRNA expression was evaluated by real-time PCR. Apoptosis was also evaluated by TUNEL assay and cell proliferation identified using an antibody against proliferating cell nuclear antigen (PCNA). The copper concentrations used did not induce the upregulation of caspase-3 gene in O. niloticus gill. In addition, in the gills of fish exposed to copper there was no increase in the estimated relative volume of apoptotic cells, indicating that neither the caspase-3-dependent or caspase-independent apoptotic pathways were induced. On the other hand, the increase in the volumetric density of epithelial proliferating cells suggests a concentration-dependent repair response.

  3. Hollow fiber cell fishing with high performance liquid chromatography for screening bioactive compounds from traditional Chinese medicines.

    Science.gov (United States)

    Xue, Xue; Li, Lihua; Chen, Xuan; Hu, Shuang; Bai, Xiaohong

    2013-03-08

    A novel hollow fiber cell fishing method with high performance liquid chromatography was proposed and used to screen, isolate, and analyze bioactive compounds from Traditional Chinese Medicines (TCMs). The active compounds that interact with the living cells acceptor inside the hollow fiber lumen were screened and isolated from the TCM extracts in phosphate buffer solution (pH 7.4). Subsequently, the active compounds bound to the cells were desorbed with methanol, and were analyzed using HPLC. HFCF with HPLC was introduced for the screening and analysis of lignans in Schisandra chinensis (Turcz) Baill and coumarins in Fructus Cnidii and Fructus Psoraleae. The surface properties of the hollow fibers filled with living cells were characterized. The nonspecific binding between the active centers of the hollow fibers and the bioactive compounds were investigated. The cell survival rates were determined before and after the screening. The repeatability of the method was tested. Some structures of the lignans and coumarins screened from TCMs were identified by the comparison to the retention times of the reference substances. HFCF-HPLC is a simple, fast, effective, and reliable method for the screening and analysis of bioactive compounds, and it can be extended to screen other bioactive compounds from TCMs.

  4. Toxic effects of mining effluents on fish gills in a subarctic lake system in NW Russia.

    Science.gov (United States)

    Tkatcheva, Victoria; Hyvärinen, Heikki; Kukkonen, Jussi; Ryzhkov, Leonid P; Holopainen, Ismo J

    2004-03-01

    The mining company Karelian Pellet in northwestern Russia extracts iron ore and processes it locally into pellets. The production operations affect the environment in the form of air pollution and wastewater emissions to lakes downstream from the factory. The toxic effects of the mining effluents on gills of perch (Perca fluviatilis) and roach (Rutilus rutilus) were studied. In lakes close to the factory, heavy metal concentrations in gills were not high, even though the metal content in sediment was elevated. In fish gills the relative proportion of phosphatidylcholine was elevated and cholesterol reduced, while the histological structure of the gills was changed. The number of mucus cells, as well as the sizes and the lengths of open areas in the chloride cells, had increased in spring and summer. The hypertrophy of chloride cells is possibly caused by the increased ambient concentrations of K+ and Li+. Changes in gill cholesterol and phospholipid proportions increase the fluidity of membranes and possibly strengthen their protective qualities, counterbalancing the adverse changes in chloride cell structure. The bioavailability and toxic effects of metals on fish are reduced by the hardness and high pH of water discharged by the mining plant.

  5. 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.

  6. 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)

  7. 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.

  8. Rh protein expression in branchial neuroepithelial cells, and the role of ammonia in ventilatory control in fish.

    Science.gov (United States)

    Zhang, Li; Michele Nawata, C; De Boeck, Gudrun; Wood, Chris M

    2015-08-01

    Bill Milsom has made seminal contributions to our understanding of ventilatory control in a wide range of vertebrates. Teleosts are particularly interesting, because they produce a 3rd, potentially toxic respiratory gas (ammonia) in large amounts. Fish are well known to hyperventilate under high environmental ammonia (HEA), but only recently has the potential role of ammonia in normal ventilatory control been investigated. It is now clear that ammonia can act directly as a ventilatory stimulant in trout, independent of its effects on acid-base balance. Even in ureotelic dogfish sharks, acute elevations in ammonia cause increases in ventilation. Peripherally, the detection of elevated ammonia resides in gill arches I and II in trout, and in vitro, neuroepithelial cells (NECs) from these arches are sensitive to ammonia, responding with elevations in intracellular Ca(2+) ([Ca(2+)]i). Centrally, hyperventilatory responses to ammonia correlate more closely with concentrations of ammonia in the brain than in plasma or CSF. After chronic HEA exposure, ventilatory responsiveness to ammonia is lost, associated with both an attenuation of the [Ca(2+)]i response in NECs, and the absence of elevation in brain ammonia concentration. Chronic exposure to HEA also causes increases in the mRNA expression of several Rh proteins (ammonia-conductive channels) in both brain and gills. "Single cell" PCR techniques have been used to isolate the individual responses of NECs versus other gill cell types. We suggest several circumstances (post-feeding, post-exercise) where the role of ammonia as a ventilatory stimulant may have adaptive benefits for O2 uptake in fish.

  9. Fish allergy and fish allergens

    DEFF Research Database (Denmark)

    Kuehn, A; Hilger, Christiane; Ollert, Markus

    2016-01-01

    Fish is one of the main elicitors for food allergies. For a long time, the clinical picture of fish allergy was reduced to the following features. First, fish-allergic patients suffer from a high IgE cross-reactivity among fishes so that they have to avoid all species. Second, clinically relevant...... review gives an overview on the clinical characteristics of fish allergy and the molecular properties of relevant fish allergens. The advancement of the IgE-based diagnosis using a panel of well-defined fish allergens from different species is in the focus of the discussion. © 2016 Dustri-Verlag Dr. Karl...

  10. 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.

  11. 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; 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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.

  12. 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...

  13. Diversity, molecular characterization and expression of T cell receptor γ in a teleost fish, the sea bass (Dicentrarchus labrax, L.

    Directory of Open Access Journals (Sweden)

    Francesco Buonocore

    Full Text Available Two lineages of T cells, expressing either the αβ T cell receptor (TR or the γδ TR, exist in Gnathostomes. The latter type of T cells account for 1-10 % of T cells in blood and up to 30 % in the small intestine. They may recognize unconventional antigens (phosphorylated microbial metabolites, lipid antigens without the need of major histocompatibility class I (MH1 or class II (MH2 presentation. In this work we have described cloning and structural characterization of TR -chain (TRG from the teleost Dicentrarchus labrax. Further, by means of quantitative PCR analysis, we analyzed TRG expression levels both in poly I:C stimulated leukocytes in vitro, and following infection with betanodavirus in vivo. Two full length cDNAs relative to TRG, with the highest peptide and nucleotide identity with Japanese flounder, were identified. A multiple alignment analysis showed the conservation of peptides fundamental for TRG biological functions, and of the FGXG motif in the FR4 region, typical of most TR and immunoglobulin light chains. A 3D structure consisting of two domains mainly folded as beta strands with a sandwich architecture for each domain was also reported. TRG CDR3 of 8-18 AA in length and diversity in the TRG rearrangements expressed in thymus and intestine for a given V/C combination were evidenced by junction length spectratyping. TRG mRNA expression levels were high in basal conditions both in thymus and intestine, while in kidney and gut leukocytes they were up-regulated after in vitro stimulation by poly I:C. Finally, in juveniles the TRG expression levels were up-regulated in the head kidney and down-regulated in intestine after in vivo infection with betanodavirus. Overall, in this study the involvement of TRG-bearing T cells during viral stimulation was described for the first time, leading to new insights for the identification of T cell subsets in fish.

  14. Antarctic Fishes.

    Science.gov (United States)

    Eastman, Joseph T.; DeVries, Arthur L.

    1986-01-01

    Explains the adaptations to Antarctic waters that Notothenioidei, a group of advanced bony fishes, have exhibited. Discusses the fishes' mechanisms of production of antifreeze properties and their capacities for neutral buoyancy in water. (ML)

  15. 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.

  16. NHE-1 sequence and expression in toad, snake and fish red blood cells

    DEFF Research Database (Denmark)

    Thomsen, Steffen Nyegaard; Wang, Tobias; Kristensen, Torsten

    Red blood cells (RBC) from reptiles appear not to express regulatory volume increase (RVI) upon shrinkage (Kristensen et al., 2008). In other vertebrates, the RVI response is primarily mediated by activation of the Na+/H+ exchanger (NHE-1) and we, therefore decided to investigate whether red cell...

  17. First identification of regulatory B cell subsets expressing IL-10 in teleost fish

    Science.gov (United States)

    IL-10 is an immunoregulatory cytokine with a potent anti-inflammatory activity, thus inhibiting the production of proinflammatory cytokines as well as processes of antigen presentation. IL-10 is produced by variety of cells, including antigen presentation cells (i.e., monocytes, macrophages and den...

  18. A sensitive fluorescent sensor for the detection of endogenous hydroxyl radicals in living cells and bacteria and direct imaging with respect to its ecotoxicity in living zebra fish.

    Science.gov (United States)

    Liu, Fei; Du, Juan; Song, Da; Xu, Meiying; Sun, Guoping

    2016-03-28

    We have synthesized a novel fluorescent probe, , which shown a high potential for imaging of endogenous ˙OH in living cells and various types of bacteria. In addition, it is an excellent sensor for in vivo imaging of ˙OH generated following treatment with TiO2NPs in zebra fish.

  19. Role of immunohistochemistry and fluorescence in-situ hybridization (FISH in the diagnosis of spindle and round cell tumors of the kidney

    Directory of Open Access Journals (Sweden)

    M. Abbas

    2015-09-01

    Conclusion: In summary we advise an immunohistochemical panel for round/spindle cell tumors of the kidney and for unclear cases we advise to add (FISH to get the correct diagnosis, as they are completely different regarding surgical approach and post-operative adjuvant therapy.

  20. The goitrogen 6-n-propyl-2-thiouracil (PTU) given during testis development increases Sertoli and germ cell numbers per cyst in fish: the tilapia (Oreochromis niloticus) model.

    Science.gov (United States)

    Matta, Sérgio L P; Vilela, Daniel A R; Godinho, Hugo P; França, Luiz R

    2002-03-01

    The main objectives of the present study were to investigate the effects of 6-n-propyl-2-thiouracil (PTU) on Sertoli cell proliferation, germ cell number, and testis size in Nile tilapias (Oreochromis niloticus). In this regard, young fish (approximately 1 g BW and approximately 3.5 cm total in length) were treated for a period of 40 d with different concentrations (100 and 150 ppm) of PTU. The animals were killed and analyzed on d 1, 30, 40, 98, and 208 after the beginning of the treatment. On d 30 and 40 the spermatogenic process was delayed in fish treated with PTU compared with the control group. Also at these periods, treated tilapia had decreased (P PTU-treated fish and were similar (P > 0.05) to those of the controls. However, testis weight and gonadosomatic index (testis mass/body weight) were approximately 100% higher (P PTU treatment. Compared with controls, at 208 d all parameters analyzed presented the same trend as that observed at 98 d. In general, at 98 d the different PTU concentrations used during the treatment period induced similar effects. However, at 208 d the mean values observed for several parameters were significantly higher (P 0.05) in the three groups of fish investigated. Remarkably, the results found in tilapia were similar to those found for rats treated with PTU. This suggests strongly that the mechanisms of control of Sertoli cell and Leydig cell proliferation seem to be preserved during vertebrate evolution.

  1. SUPPLEMENTATION OF PATIENTS WITH HOMOZYGOUS SICKLE-CELL DISEASE WITH ZINC, ALPHA-TOCOPHEROL, VITAMIN-C, SOYBEAN OIL, AND FISH OIL

    NARCIS (Netherlands)

    MUSKIET, FAJ; MUSKIET, FD; MEIBORG, G; SCHERMER, JG

    1991-01-01

    Thirteen patients (aged 0.7-17.9 y) with homozygous sickle cell disease were supplemented with alpha-tocopherol, vitamin C, zinc, and soybean oil (suppl 1; for 8 mo) and alpha-tocopherol, vitamin C, and fish oil (suppl 2; for 7 mo). Urinary zinc (suppl 1), plasma vitamin C, plasma cholesterol ester

  2. 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.

  3. 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...

  4. Fish Dishes.

    Science.gov (United States)

    Derby, Marie

    2003-01-01

    Describes an art project that was inspired by Greek pottery, specifically dishes shaped as fish. Explains that fourth-grade students drew a fish shape that was later used to create their clay version of the fish. Discusses how the students examined the pottery to make decisions about color and design. (CMK)

  5. Dietary fish oil reverse epididymal tissue adiposity, cell hypertrophy and insulin resistance in dyslipemic sucrose fed rat model small star, filled.

    Science.gov (United States)

    Soria, Ana; Chicco, Adriana; Eugenia D'Alessandro, María; Rossi, Andrea; Lombardo, Yolanda B.

    2002-04-01

    The present work was designed to assess the possible benefits of (7% w/w) dietary fish oil in reversing the morphological and metabolic changes present in the adipose tissue of rats fed an SRD for a long time. With this purpose, in the epididymal fat tissue, we investigated the effect of dietary fish oil upon: i) the number, size and distribution of cells, ii) the basal and stimulated lipolysis, iii) the lipoprotein lipase (LPL) and the glucose 6-phosphate dehydrogenase activities, and iv) the antilipolytic action of insulin. The study was conducted on rats fed an SRD during 120 days with fish oil being isocaloric substituted for corn oil for 90-120 days in half the animals. Permanent hypertriglyceridemia, insulin resistance and abnormal glucose homeostasis were present in the rats before the source of fat in the diet was replaced. The major new findings of this study are the following: i) Dietary fish oil markedly reduced the fat pads mass, the hypertrophy of fat cells and improved the altered cell size distribution. ii) The presence of fish oil in the diet corrected the inhibitory effect of high sucrose diet upon the antilipolytic action of insulin, reduced the "in vitro" enhanced basal lipolysis and normalized isoproterenol-stimulated lipolysis. Fat pads lipoprotein lipase activity decreased reaching values similar to those observed in age-matched controls fed a control diet (CD). These effects were not accompanied by any change in rat body weight. All these data suggest that the dyslipemic rats fed a moderate amount of dietary fish oil constitute a useful animal model to study diet-regulated insulin action.

  6. BIOTECHNOLOGY OF THE FISH AQUACULTURE

    Directory of Open Access Journals (Sweden)

    L. P. Buchatsky

    2013-12-01

    Full Text Available The latest progress in biotechnology on fish aquaculture and different modern methods of investigations for increasing of fish productivity in aquaculture are analyzed. Except for the applied aspect, the use of modern biotechnological methods of investigations opens new possibilities for fundamental researches of sex-determining mechanisms, polyploidy, distant hybridization, and developmental biology of bony fishes. Review contains examples of utilizing modern biotechnology methods to obtain transgenic fishes with accelerated growth and for designing surrogate fishes. Methods for receiving unisexual shoals of salmon and sturgeon female fishes with the view of obtaining a large quantity of caviar, as well as receiving sterile (triploid fishes are analyzed. Great attention is given to androgenesis, particularly to disperm one, in connection with the problem of conserving rare and vanishing fish species using only sperm genetic material. Examples how distant hybrids may be obtained with the use of disperm androgenesis and alkylated DNA are given. Methods of obtaining fish primordium germ cells, recent developments in cultivation of fish stem cells and their use in biotechnology, as well as ones of transplantation of oogonium and spermatogonium to obtain surrogate fishes. The examples of successful experiments on spermatogonial xenotransplantation and characteristic of antifreezing fish proteins and also the prospect of their practical usage are given.

  7. Inorganic mercury accumulation in brain following waterborne exposure elicits a deficit on the number of brain cells and impairs swimming behavior in fish (white seabream-Diplodus sargus).

    Science.gov (United States)

    Pereira, Patrícia; Puga, Sónia; Cardoso, Vera; Pinto-Ribeiro, Filipa; Raimundo, Joana; Barata, Marisa; Pousão-Ferreira, Pedro; Pacheco, Mário; Almeida, Armando

    2016-01-01

    The current study contributes to fill the knowledge gap on the neurotoxicity of inorganic mercury (iHg) in fish through the implementation of a combined evaluation of brain morphometric alterations (volume and total number of neurons plus glial cells in specific regions of the brain) and swimming behavior (endpoints related with the motor activity and mood/anxiety-like status). White seabream (Diplodus sargus) was exposed to realistic levels of iHg in water (2μgL(-1)) during 7 (E7) and 14 days (E14). After that, fish were allowed to recover for 28 days (PE28) in order to evaluate brain regeneration and reversibility of behavioral syndromes. A significant reduction in the number of cells in hypothalamus, optic tectum and cerebellum was found at E7, accompanied by relevant changes on swimming behavior. Moreover, the decrease in the number of neurons and glia in the molecular layer of the cerebellum was followed by a contraction of its volume. This is the first time that a deficit on the number of cells is reported in fish brain after iHg exposure. Interestingly, a recovery of hypothalamus and cerebellum occurred at E14, as evidenced by the identical number of cells found in exposed and control fish, and volume of cerebellum, which might be associated with an adaptive phenomenon. After 28 days post-exposure, the optic tectum continued to show a decrease in the number of cells, pointing out a higher vulnerability of this region. These morphometric alterations coincided with numerous changes on swimming behavior, related both with fish motor function and mood/anxiety-like status. Overall, current data pointed out the iHg potential to induce brain morphometric alterations, emphasizing a long-lasting neurobehavioral hazard.

  8. 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...

  9. 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.

  10. 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.

  11. Toxicological assessment of indium nitrate on aquatic organisms and investigation of the effects on the PLHC-1 fish cell line

    Energy Technology Data Exchange (ETDEWEB)

    Zurita, Jorge L. [National Institute of Toxicology and Forensic Sciences, Av. Dr Fedriani s/n, 41009, Seville (Spain); Jos, Angeles [Area of Toxicology, University of Seville, Prof. Garcia Gonzalez 2, 41012, Seville (Spain); Peso, Ana del; Salguero, Manuel [National Institute of Toxicology and Forensic Sciences, Av. Dr Fedriani s/n, 41009, Seville (Spain); Camean, Ana M. [Area of Toxicology, University of Seville, Prof. Garcia Gonzalez 2, 41012, Seville (Spain); Lopez-Artiguez, Miguel [National Institute of Toxicology and Forensic Sciences, Av. Dr Fedriani s/n, 41009, Seville (Spain); Repetto, Guillermo [National Institute of Toxicology and Forensic Sciences, Av. Dr Fedriani s/n, 41009, Seville (Spain); Area of Toxicology, University of Seville, Prof. Garcia Gonzalez 2, 41012, Seville (Spain)], E-mail: repetto@us.es

    2007-11-15

    Indium nitrate is mainly used as a semiconductor in batteries, for plating and other chemical and medical applications. There is a lack of available information about the adverse effects of indium compounds on aquatic organisms. Therefore, the toxic effects on systems from four trophic levels of the aquatic ecosystem were investigated. Firstly, the bacterium Vibrio fischeri, the alga Chlorella vulgaris and the cladoceran Daphnia magna were used in the toxicological evaluation of indium nitrate. The most sensitive model was V. fischeri, with a NOAEL of 0.02 and an EC{sub 50} of 0.04 mM at 15 min. Although indium nitrate should be classified as harmful to aquatic organisms, it is not expected to represent acute risk to the aquatic biota. Secondly, PLHC-1 fish cell line was employed to investigate the effects and mechanisms of toxicity. Although protein content, neutral red uptake, methylthiazol metabolization, lysosomal function and acetylcholinesterase activity were reduced in cells, stimulations were observed for metallothionein levels and succinate dehydrogenase and glucose-6-phosphate dehydrogenase activities. No changes were observed in ethoxyresorufin-O-deethylase activity. To clarify the main events in PLHC-1 cell death induced by indium nitrate, nine modulators were applied. They were related to oxidative stress ({alpha}-tocopherol succinate, mannitol and sodium benzoate), disruption of calcium homeostasis (BAPTA-AM and EGTA), thiol protection (1,4-dithiotreitol), iron chelation (deferoxiamine) or regulation of glutathione levels (2-oxothiazolidine-4-carboxylic acid and malic acid diethyl ester). The main morphological alterations were hydropic degeneration and loss of cells. At least, in partly, toxicity seems to be mediated by oxidative stress, and particularly by NADPH-dependent lipid peroxidation.

  12. Toxicological assessment of indium nitrate on aquatic organisms and investigation of the effects on the PLHC-1 fish cell line.

    Science.gov (United States)

    Zurita, Jorge L; Jos, Angeles; del Peso, Ana; Salguero, Manuel; Cameán, Ana M; López-Artíguez, Miguel; Repetto, Guillermo

    2007-11-15

    Indium nitrate is mainly used as a semiconductor in batteries, for plating and other chemical and medical applications. There is a lack of available information about the adverse effects of indium compounds on aquatic organisms. Therefore, the toxic effects on systems from four trophic levels of the aquatic ecosystem were investigated. Firstly, the bacterium Vibrio fischeri, the alga Chlorella vulgaris and the cladoceran Daphnia magna were used in the toxicological evaluation of indium nitrate. The most sensitive model was V. fischeri, with a NOAEL of 0.02 and an EC(50) of 0.04 mM at 15 min. Although indium nitrate should be classified as harmful to aquatic organisms, it is not expected to represent acute risk to the aquatic biota. Secondly, PLHC-1 fish cell line was employed to investigate the effects and mechanisms of toxicity. Although protein content, neutral red uptake, methylthiazol metabolization, lysosomal function and acetylcholinesterase activity were reduced in cells, stimulations were observed for metallothionein levels and succinate dehydrogenase and glucose-6-phosphate dehydrogenase activities. No changes were observed in ethoxyresorufin-O-deethylase activity. To clarify the main events in PLHC-1 cell death induced by indium nitrate, nine modulators were applied. They were related to oxidative stress (alpha-tocopherol succinate, mannitol and sodium benzoate), disruption of calcium homeostasis (BAPTA-AM and EGTA), thiol protection (1,4-dithiotreitol), iron chelation (deferoxiamine) or regulation of glutathione levels (2-oxothiazolidine-4-carboxylic acid and malic acid diethyl ester). The main morphological alterations were hydropic degeneration and loss of cells. At least, in partly, toxicity seems to be mediated by oxidative stress, and particularly by NADPH-dependent lipid peroxidation.

  13. Rapid oriented fibril formation of fish scale collagen facilitates early osteoblastic differentiation of human mesenchymal stem cells.

    Science.gov (United States)

    Matsumoto, Rena; Uemura, Toshimasa; Xu, Zhefeng; Yamaguchi, Isamu; Ikoma, Toshiyuki; Tanaka, Junzo

    2015-08-01

    We studied the effect of fibril formation of fish scale collagen on the osteoblastic differentiation of human mesenchymal stem cells (hMSCs). We found that hMSCs adhered easily to tilapia scale collagen, which remarkably accelerated the early stage of osteoblastic differentiation in hMSCs during in vitro cell culture. Osteoblastic markers such as ALP activity, osteopontin, and bone morphogenetic protein 2 were markedly upregulated when the hMSCs were cultured on a tilapia collagen surface, especially in the early osteoblastic differentiation stage. We hypothesized that this phenomenon occurs due to specific fibril formation of tilapia collagen. Thus, we examined the time course of collagen fibril formation using high-speed atomic force microscopy. Moreover, to elucidate the effect of the orientation of fibril formation on the differentiation of hMSCs, we measured ALP activity of hMSCs cultured on two types of tilapia scale collagen membranes with different degrees of fibril formation. The ALP activity in hMSCs cultured on a fibrous collagen membrane was significantly higher than on a non-fibrous collagen membrane even before adding osteoblastic differentiation medium. These results showed that the degree of the fibril formation of tilapia collagen was essential for the osteoblastic differentiation of hMSCs.

  14. 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...

  15. Temperature-dependent modification of muscle precursor cell behaviour is an underlying reason for lasting effects on muscle cellularity and body growth of teleost fish.

    Science.gov (United States)

    Steinbacher, Peter; Marschallinger, Julia; Obermayer, Astrid; Neuhofer, Alois; Sänger, Alexandra M; Stoiber, Walter

    2011-06-01

    Temperature is an important factor influencing teleost muscle growth, including a lasting ('imprinted') influence of embryonic thermal experience throughout all further life. However, little is known about the cellular processes behind this phenomenon. The study reported here used digital morphometry and immunolabelling for Pax7, myogenin and H3P to quantitatively examine the effects of thermal history on muscle precursor cell (MPC) behaviour and muscle growth in pearlfish (Rutilus meidingeri) until the adult stage. Fish were reared at three different temperatures (8.5, 13 and 16°C) until hatching and subsequently kept under the same (ambient) thermal conditions. Cellularity data were combined with a quantitative analysis of Pax7+ MPCs including those that were mitotically active (Pax7+/H3P+) or had entered differentiation (Pax7+/myogenin+). The results demonstrate that at hatching, body lengths, fast and slow muscle cross-sectional areas and fast fibre numbers are lower in fish reared at 8.5 and 13°C than at 16°C. During the larval period, this situation changes in the 13°C-fish, so that these fish are finally the largest. The observed effects can be related to divergent cellular mechanisms at the MPC level that are initiated in the embryo during the imprinting period. Embryos of 16°C-fish have reduced MPC proliferation but increased differentiation, and thus give rise to larger hatchlings. However, their limited MPC reserves finally lead to smaller adults. By contrast, embryos of 13°C-fish and, to a lesser extent, 8.5°-fish, show enhanced MPC proliferation but reduced differentiation, thus leading to smaller hatchlings but allowing for a larger MPC pool that can be used for enhanced post-hatching growth, finally resulting in larger adults.

  16. Establishment of caudal fin cell lines from tropical ornamental fishes Puntius fasciatus and Pristolepis fasciata endemic to the Western Ghats of India.

    Science.gov (United States)

    Swaminathan, T Raja; Basheer, V S; Gopalakrishnan, A; Rathore, Gaurav; Chaudhary, Dharmendra K; Kumar, Raj; Jena, J K

    2013-12-01

    Two new cell lines, PFF and CFF were established from the caudal fin of the Puntius fasciatus, and Pristolepis fasciata respectively. Since their initiation, these cell lines (PFF and CFF) have been subcultured in L-15 medium with 10% fetal bovine serum for more than 35 passages at 28°C and both the cell lines were characterized. Karyotyping analysis of PFF and CFF cells at 25th passage indicated that the modal chromosome number was 2n=50 and 2n=48 respectively. The cell line was cryopreserved in liquid nitrogen at -196°C and could be recovered from storage after six months with good cell viability. Polymerase chain reaction amplification of the fragments of two mitochondrial genes, 16S rRNA and COI confirmed that the cell lines originated from the respective species. The bacterial extracellular products from Vibrio cholerae MTCC3904 and Aeromonas hydrophila were found to be toxic to PFF and CFF. Both the cells were resistant to the marine viral nervous necrosis virus (VNNV). No CPE could be found in both cell lines inoculated with the fish samples and cell culture supernatants were demonstrated free of SVC, iridovirus and KHV by molecular methods. These results indicated the absence of SVC, iridovirus and KHV in the ornamental fishes collected from the Western Ghats of India.

  17. PNA-COMBO-FISH: From combinatorial probe design in silico to vitality compatible, specific labelling of gene targets in cell nuclei.

    Science.gov (United States)

    Müller, Patrick; Rößler, Jens; Schwarz-Finsterle, Jutta; Schmitt, Eberhard; Hausmann, Michael

    2016-07-01

    Recently, advantages concerning targeting specificity of PCR constructed oligonucleotide FISH probes in contrast to established FISH probes, e.g. BAC clones, have been demonstrated. These techniques, however, are still using labelling protocols with DNA denaturing steps applying harsh heat treatment with or without further denaturing chemical agents. COMBO-FISH (COMBinatorial Oligonucleotide FISH) allows the design of specific oligonucleotide probe combinations in silico. Thus, being independent from primer libraries or PCR laboratory conditions, the probe sequences extracted by computer sequence data base search can also be synthesized as single stranded PNA-probes (Peptide Nucleic Acid probes). Gene targets can be specifically labelled with at least about 20 PNA-probes obtaining visibly background free specimens. By using appropriately designed triplex forming oligonucleotides, the denaturing procedures can completely be omitted. These results reveal a significant step towards oligonucleotide-FISH maintaining the 3d-nanostructure and even the viability of the cell target. The method is demonstrated with the detection of Her2/neu and GRB7 genes, which are indicators in breast cancer diagnosis and therapy.

  18. Genotoxicity of heterocyclic PAHs in the micronucleus assay with the fish liver cell line RTL-W1.

    Directory of Open Access Journals (Sweden)

    Markus Brinkmann

    Full Text Available Heterocyclic aromatic hydrocarbons are, together with their un-substituted analogues, widely distributed throughout all environmental compartments. While fate and effects of homocyclic PAHs are well-understood, there are still data gaps concerning the ecotoxicology of heterocyclic PAHs: Only few publications are available investigating these substances using in vitro bioassays. Here, we present a study focusing on the identification and quantification of clastogenic and aneugenic effects in the micronucleus assay with the fish liver cell line RTL-W1 that was originally derived from rainbow trout (Oncorhynchus mykiss. Real concentrations of the test items after incubation without cells were determined to assess chemical losses due to, e.g., sorption or volatilization, by means of gas chromatography-mass spectrometry. We were able to show genotoxic effects for six compounds that have not been reported in vertebrate systems before. Out of the tested substances, 2,3-dimethylbenzofuran, benzothiophene, quinoline and 6-methylquinoline did not cause substantial induction of micronuclei in the cell line. Acridine caused the highest absolute induction. Carbazole, acridine and dibenzothiophene were the most potent substances compared with 4-nitroquinoline oxide, a well characterized genotoxicant with high potency used as standard. Dibenzofuran was positive in our investigation and tested negative before in a mammalian system. Chemical losses during incubation ranged from 29.3% (acridine to 91.7% (benzofuran and may be a confounding factor in studies without chemical analyses, leading to an underestimation of the real potency. The relative potency of the investigated substances was high compared with their un-substituted PAH analogues, only the latter being typically monitored as priority or indicator pollutants. Hetero-PAHs are widely distributed in the environment and even more mobile, e.g. in ground water, than homocyclic PAHs due to the higher water

  19. 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.

  20. 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.

  1. 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

  2. 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

  3. 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…

  4. 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…

  5. 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.

  6. 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.

  7. Investigating Microbial Activity in Diazotrophic Methane Seep Sediment via Transcript Analysis and Single-Cell FISH-NanoSIMS

    Science.gov (United States)

    Dekas, A. E.; Connon, S. A.; Chadwick, G.; Orphan, V. J.

    2012-12-01

    Methane seep microbial ecosystems are phylogenetically diverse and physiologically complex, and require culture-independent techniques to accurately investigate metabolic activity. In the present study we combine an RNA analysis of four key microbial genes with FISH-NanoSIMS analysis of single cells to determine the diversity of nitrogen fixing microorganisms (diazotrophs) present at a deep-sea methane-seeping site, as well as investigate the methane-dependency of a variety of community members. Recently, methane-dependent nitrogen fixation was observed in Mound 12 Costa Rica sediments, and was spatially correlated with the abundance of aggregates of anaerobic methanotrophic archaea (ANME) and sulfate reducing bacterial symbionts (SRB). Combined with the detection of 15N uptake from 15N2 in these aggregates, this suggested that the ANME-SRB aggregates are the primary diazotrophs in seep sediment. However, the diversity of dinitrogenase reductase (nifH) sequences recovered from several deep-sea locales, including Mound 12, suggests a greater diversity of diazotrophs in marine sediment. To investigate the activity of these potential diazotrophs in Mound 12 sediment, we investigated a suite of RNA transcripts in 15N2 incubations in both the presence and absence of methane: nifH, bacterial 16S rRNA, methyl coenzyme M reductase A (mcrA), and adenosine-5'-phosposulfate reductase alpha subunit (aprA). No nifH transcripts were recovered in incubations without methane, consistent with previous measurements lacking 15N2 uptake in the same sediments. The activity of the bacterial community in general, assessed by variable transcription, was also greatly affected by the presence or absence of methane. Single-cell fluorescence in situ hybridization coupled to nanoscale secondary ion mass spectrometry (FISH-NanoSIMS) was employed to confirm diazotrophic activity (15N2 uptake) and protein synthesis (15NH4+ uptake) of particular species implicated as ecologically important by the

  8. A new and fast technique to generate offspring after germ cells transplantation in adult fish: the Nile tilapia (Oreochromis niloticus model.

    Directory of Open Access Journals (Sweden)

    Samyra M S N Lacerda

    Full Text Available BACKGROUND: Germ cell transplantation results in fertile recipients and is the only available approach to functionally investigate the spermatogonial stem cell biology in mammals and probably in other vertebrates. In the current study, we describe a novel non-surgical methodology for efficient spermatogonial transplantation into the testes of adult tilapia (O. niloticus, in which endogenous spermatogenesis had been depleted with the cytostatic drug busulfan. METHODOLOGY/PRINCIPAL FINDINGS: Using two different tilapia strains, the production of fertile spermatozoa with donor characteristics was demonstrated in adult recipient, which also sired progeny with the donor genotype. Also, after cryopreservation tilapia spermatogonial cells were able to differentiate to spermatozoa in the testes of recipient fishes. These findings indicate that injecting germ cells directly into adult testis facilitates and enable fast generation of donor spermatogenesis and offspring compared to previously described methods. CONCLUSION: Therefore, a new suitable methodology for biotechnological investigations in aquaculture was established, with a high potential to improve the production of commercially valuable fish, generate transgenic animals and preserve endangered fish species.

  9. Fish health and fish quality

    DEFF Research Database (Denmark)

    Ingerslev, Hans-Christian

    Aquaculture is an expanding worldwide industry producing an increasing amount of fish every year. The quality of the fish meat is dependent upon many biological and non-biological factors. Infectious diseases are known to cause bleedings and damage of the muscle tissue that may lead to scarring...... are poorly described in fish. The present work in this thesis focused on: 1) examination of potential changes in the quality regarding texture of the muscle tissue in rainbow trout (Oncorhynchus mykiss) after previous infection with the bacterial pathogens Yersinia ruckeri and Vibrio anguillarum; 2...... of these studies showed that previous infections by Yersinia ruckeri and Vibrio anguillarum gave rise to subsequent changes regarding textural quality parameters in fresh fish meat, while no differences were seen for cold-smoked meat from the same fish. The texture in previous infected fish was less flaky and less...

  10. Silver(I Ions Ultrasensitive Detection at Carbon Electrodes―Analysis of Waters, Tobacco Cells and Fish Tissues

    Directory of Open Access Journals (Sweden)

    Sona Krizkova

    2009-09-01

    Full Text Available We used carbon paste electrodes and a standard potentiostat to detect silver ions. The detection limit (3 Signal/Noise ratio was estimated as 0.5 μM. A standard electrochemical instrument microanalysis of silver(I ions was suggested. As a working electrode a carbon tip (1 mL or carbon pencil was used. Limits of detection estimated by dilution of a standard were 1 (carbon tip or 10 nM (carbon pencil. Further we employed flow injection analysis coupled with carbon tip to detect silver(I ions released in various beverages and mineral waters. During first, second and third week the amount of silver(I ions releasing into water samples was under the detection limit of the technique used for their quantification. At the end of a thirteen weeks long experiment the content of silver(I ions was several times higher compared to the beginning of release detected in the third week and was on the order of tens of nanomoles. In subsequent experiments the influence of silver(I ions (0, 5 and 10 μM on a plant model system (tobacco BY-2 cells during a fourday exposition was investigated. Silver(I ions were highly toxic to the cells, which was revealed by a double staining viability assay. Moreover we investigated the effect of silver(I ions (0, 0.3, 0.6, 1.2 and 2.5 μM on guppies (Poecilia reticulata. Content of Ag(I increased with increasing time of the treatment and applied concentrations in fish tissues. It can be concluded that a carbon tip or carbon pencil coupled with a miniaturized potentiostat can be used for detection of silver(I ions in environmental samples and thus represents a small, portable, low cost and easy-to-use instrument for such purposes.

  11. 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.

  12. Fish parasites

    DEFF Research Database (Denmark)

    This book contains 22 chapters on some of the most important parasitic diseases in wild and farmed fish. International experts give updated reviews and provide solutions to the problems......This book contains 22 chapters on some of the most important parasitic diseases in wild and farmed fish. International experts give updated reviews and provide solutions to the problems...

  13. 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...

  14. 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...

  15. Medaka haploid embryonic stem cells are susceptible to Singapore grouper iridovirus as well as to other viruses of aquaculture fish species.

    Science.gov (United States)

    Yuan, Yongming; Huang, Xiaohong; Zhang, Lei; Zhu, Yi; Huang, Youhua; Qin, Qiwei; Hong, Yunhan

    2013-10-01

    Viral infection is a challenge in high-density aquaculture, as it leads to various diseases and causes massive or even complete loss. The identification and disruption of host factors that viruses utilize for infection offer a novel approach to generate viral-resistant seed stocks for cost-efficient and sustainable aquaculture. Genetic screening in haploid cell cultures represents an ideal tool for host factor identification. We have recently generated haploid embryonic stem (ES) cells in the laboratory fish medaka. Here, we report that HX1, one of the three established medaka haploid ES cell lines, was susceptible to the viruses tested and is thus suitable for genetic screening to identify host factors. HX1 cells displayed a cytopathic effect and massive death upon inoculation with three highly infectious and notifiable fish viruses, namely Singapore grouper iridovirus (SGIV), spring viremia of carp virus (SVCV) and red-spotted grouper nervous necrosis virus (RGNNV). Reverse transcription-PCR and Western blot analyses revealed the expression of virus genes. SGIV infection in HX1 cells elicited a host immune response and apoptosis. Viral replication kinetics were determined from a virus growth curve, and electron microscopy revealed propagation, assembly and release of infectious SGIV particles in HX1 cells. Our results demonstrate that medaka haploid ES cells are susceptible to SGIV, as well as to SVCV and RGNNV, offering a unique opportunity for the identification of host factors by genetic screening.

  16. Role of pathogen-derived cell wall carbohydrates and prostaglandin E2 in immune response and suppression of fish immunity by the oomycete Saprolegnia parasitica.

    Science.gov (United States)

    Belmonte, Rodrigo; Wang, Tiehui; Duncan, Gary J; Skaar, Ida; Mélida, Hugo; Bulone, Vincent; van West, Pieter; Secombes, Christopher J

    2014-11-01

    Saprolegnia parasitica is a freshwater oomycete that is capable of infecting several species of fin fish. Saprolegniosis, the disease caused by this microbe, has a substantial impact on Atlantic salmon aquaculture. No sustainable treatment against saprolegniosis is available, and little is known regarding the host response. In this study, we examined the immune response of Atlantic salmon to S. parasitica infection and to its cell wall carbohydrates. Saprolegnia triggers a strong inflammatory response in its host (i.e., induction of interleukin-1β1 [IL-1β1], IL-6, and tumor necrosis factor alpha), while severely suppressing the expression of genes associated with adaptive immunity in fish, through downregulation of T-helper cell cytokines, antigen presentation machinery, and immunoglobulins. Oomycete cell wall carbohydrates were recognized by fish leukocytes, triggering upregulation of genes involved in the inflammatory response, similar to what is observed during infection. Our data suggest that S. parasitica is capable of producing prostaglandin [corrected] E2 (PGE2) in vitro, a metabolite not previously shown to be produced by oomycetes, and two proteins with homology to vertebrate enzymes known to play a role in prostaglandin biosynthesis have been identified in the oomycete genome. Exogenous PGE2 was shown to increase the inflammatory response in fish leukocytes incubated with cell wall carbohydrates while suppressing genes involved in cellular immunity (gamma interferon [IFN-γ] and the IFN-γ-inducible protein [γ-IP]). Inhibition of S. parasitica zoospore germination and mycelial growth by two cyclooxygenase inhibitors (aspirin and indomethacin) also suggests that prostaglandins may be involved in oomycete development.

  17. Flow cytometric detection of micronuclei and cell cycle alterations in fish-derived cells after exposure to three model genotoxic agents: mitomycin C, vincristine sulfate and benzo(a)pyrene.

    Science.gov (United States)

    Sánchez, P; Llorente, M T; Castaño, A

    2000-02-16

    The measurement of cytogenetic alterations in vitro is considered an initial step in the risk assessment procedures for genotoxic agents. The concern about genotoxic pollutants in natural fish population makes the use of fish-derived cells an useful tool for these purposes. The technological improvements in well-established cytogenetic endpoints, such as micronuclei (MN) estimations by means of flow cytometry, have been proposed in the later years using mammalian cells. In this work, we test the capability of flow cytometry to evaluate MN induction and cell cycle alterations in an established fish cell line (RTG-2) using three agent-inductor models at different concentrations and exposure periods. For mitomycin C, an inverse relationship between length of exposure period and concentrations was observed. A dose-response relationship was observed after exposing RTG-2 cells to vincristine sulfate and benzo(a)pyrene. As this study shows, RTG-2 cells respond to clastogenic and aneugenic effects of the tested chemicals through the induction of MN at similar doses to mammalian cells and without the addition of exogenous metabolic activity. The possibility to check cell cycle alterations, in the same sample, gives the opportunity to evaluate early signals of cytotoxicity. The use of flow cytometry improves the assay by means of its speed and objectivity, which makes the assay very useful for genotoxicity assessment of aquatic chemicals.

  18. 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

  19. 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...

  20. 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...

  1. 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...

  2. 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...

  3. 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)...

  4. Immunity to Fish Rhabdoviruses

    Directory of Open Access Journals (Sweden)

    Maureen K. Purcell

    2012-01-01

    Full Text Available Members of the family Rhabdoviridae are single-stranded RNA viruses and globally important pathogens of wild and cultured fish and thus relatively well studied in their respective hosts or other model systems. Here, we review the protective immune mechanisms that fish mount in response to rhabdovirus infections. Teleost fish possess the principal components of innate and adaptive immunity found in other vertebrates. Neutralizing antibodies are critical for long-term protection from fish rhabdoviruses, but several studies also indicate a role for cell-mediated immunity. Survival of acute rhabdoviral infection is also dependent on innate immunity, particularly the interferon (IFN system that is rapidly induced in response to infection. Paradoxically, rhabdoviruses are sensitive to the effects of IFN but virulent rhabdoviruses can continue to replicate owing to the abilities of the matrix (M protein to mediate host-cell shutoff and the non‑virion (NV protein to subvert programmed cell death and suppress functional IFN. While many basic features of the fish immune response to rhabdovirus infections are becoming better understood, much less is known about how factors in the environment affect the ecology of rhabdovirus infections in natural populations of aquatic animals.

  5. 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.

  6. 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

  7. 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)

  8. 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.

  9. In vivo and in vitro mixed-function oxygenase activity and vitellogenin induction in fish and in fish and rat liver cells by stilbenes isolated from scotch pine (Pinus sylvestris).

    Science.gov (United States)

    Parrott, Joanne L; Kohli, Jagmohan; Sherry, James P; Hewitt, L Mark

    2011-01-01

    Many types of pulp and paper mill effluents have the ability to induce mixed-function oxygenase (MFO) activity and vitellogenin (VTG) protein in exposed male fish. The search for the compounds responsible for MFO induction has led to several classes of compounds, among them retene and stilbenes. The objective of this study was to investigate the biological activities of candidate stilbene compounds. Three stilbenes, 3,5-dihydroxystilbene (pinosylvin; P1), 3-hydroxy-5-methoxystilbene (P2), and 3,5-dimethoxystilbene (P3), were extracted from Scotch pine (Pinus sylvestris) and purified to evaluate their ability to induce MFO activity in vitro using ethoxyresorufin-O-deethylase (EROD) activity in a rat hepatoma cell line (H4IIE). As these compounds may be chlorinated during pulp bleaching, chlorination of P2 was undertaken, producing di- and trichlorinated isomers (Cl-P2), which were also tested. Compounds were tested for EROD-inducing ability in vivo by exposing juvenile rainbow tout (Oncorhynchus mykiss) to waterborne concentrations (0.010 to 1.0 mg/L) for 4 days. Compounds were also tested for their ability to induce VTG in trout primary liver cells in vitro. The stilbenes were weak inducers of EROD and VTG. H4IIE EROD was induced by all four compounds, with the most potent induction by P3, followed by P1, the Cl-P2 mixture, and then P2. Induction for all four stilbenes was from 3.13 × 10⁻³ to 3.57 × 10⁻⁴ as potent as retene and about 1.11 × 10⁻⁵ to 1.20 × 10⁻⁶ as potent as TCDD. Juvenile rainbow trout did not show EROD induction after exposures to P1, P2, or the Cl-P2 mixture, whereas P3 caused activity fourfold above that of controls. P1, P3, and Cl-P2 all weakly induced VTG in rainbow trout hepatocytes. The most potent inducer of VTG was Cl-P2, followed by P3 and P1. The results show the ability of wood-derived stilbenes to cause weak MFO induction in fish and in rat liver cells and to weakly induce vitellogenin in fish liver cells.

  10. 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.

  11. 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.

  12. 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...

  13. One Fish, Two Fish, Redfish, You Fish!

    Science.gov (United States)

    White, Katherine; Timmons, Maryellen; Medders, Paul

    2011-01-01

    The recreational fishing activity presented in this article provides a hands-on, problem-based experience for students; it unites biology, math, economics, environmental policy, and population dynamics concepts. In addition, the activity allows students to shape environmental policy in a realistic setting and evaluate their peers' work. By…

  14. Neuroglial cells in long-term primary cultures from the gilthead sea bream (Sparus aurata L.: new functional in vitro model from bony fish brain

    Directory of Open Access Journals (Sweden)

    Gerardo Centoducati

    2013-01-01

    Full Text Available Neuroglia has been historically considered the “glue” of the nervous system, as the ancient Greek name suggests, being simply referred as non-neuronal cells, with supporting functions for neurons in the CNS of mammalian and lower vertebrates. All around the world, approximately 283 cell lines were obtained from fish, yet none of these was from the brain of Sparus aurata, neither in cell lines nor as primary culture. Here we describe a novel in vitro reproducible neuroglial marine model for establishing primary neuroglial cell cultures, by dissociating the whole brain of seabream juveniles. We showed that proliferating neural stem cells produced alongside three generating lineages, such as neuronal precursor cells, astroglial precursor cells and oligodendroglia precursor cells, which developed respectively neurons, astrocytes and oligodendrocytes. The radial glia, finely described by morphological studies and immunochemical antigen expression, showed a peculiar spatial distribution, giving rise simultaneously both to astrocytes and neuronal precursors within a highly proliferative assemblate. Radial glia cells were assessed by glial fibrillary acidic protein (GFAP and vimentin reactivity, astrocytes by GFAP, neurons by the neuron-specific markers for ubiquitin carboxy-terminal hydrolase 1 (UCHL1 and intermediate filament associated protein (NF, whereas myelinating oligodendrocytes were immunostained with anti-myelin basic protein (MBP and anti-O4. Our findings suggest that seabream neuroglial cells gain in 3-4 weeks of culturing proliferation, neuroglial differentiation, and oligodendrocyte maturation with myelination, thus disclosing on the possibility that mixed neuroglial cultures can accelerate the maturation of oligodendrocytes and the regeneration of CNS injury in fish.

  15. Cytogenetic Characterization of the TM4 Mouse Sertoli Cell Line. II. Chromosome Microdissection, FISH, Scanning Electron Microscopy, and Confocal Laser Scanning Microscopy.

    Science.gov (United States)

    Schmid, Michael; Guttenbach, Martina; Steinlein, Claus; Wanner, Gerhard; Houben, Andreas

    2015-01-01

    The chromosomes and interphase cell nuclei of the permanent mouse Sertoli cell line TM4 were examined by chromosome microdissection, FISH, scanning electron microscopy, and confocal laser scanning microscopy. The already known marker chromosomes m1-m5 were confirmed, and 2 new large marker chromosomes m6 and m7 were characterized. The minute heterochromatic marker chromosomes m4 and m5 were microdissected and their DNA amplified by DOP-PCR. FISH of this DNA probe on TM4 metaphase chromosomes demonstrated that the m4 and m5 marker chromosomes have derived from the centromeric regions of normal telocentric mouse chromosomes. Ectopic pairing of the m4 and m5 marker chromosomes with the centromeric region of any of the other chromosomes (centromeric associations) was apparent in ∼60% of the metaphases. Scanning electron microscopy revealed DNA-protein bridges connecting the centromeric regions of normal chromosomes and the associated m4 and m5 marker chromosomes. Interphase cell nuclei of TM4 Sertoli cells did not exhibit the characteristic morphology of Sertoli cells in the testes of adult mice as shown by fluorescence microscopy and confocal laser scanning microscopy.

  16. Fighting fish

    Science.gov (United States)

    Duchi, E.; Guerrini, V.; Rinaldi, S.; Schaeffer, G.

    2017-01-01

    We introduce new combinatorial structures, called fighting fish, that generalize directed convex polyominoes by allowing them to branch out of the plane into independent substructures. On the one hand the combinatorial structure of fighting fish appears to be particularly rich: we show that their generating function with respect to the perimeter and number of tails is algebraic, and we conjecture a mysterious multivariate equidistribution property with the left ternary trees introduced by Del Lungo et al On the other hand, fighting fish provide a simple and natural model of random branching surfaces which displays original features: in particular, we show that the average area of a uniform random fighting fish with perimeter 2n is of order n 5/4: to the best of our knowledge this behaviour is non-standard and suggests that we have identified a new universality class of random structures. Dedicated to Tony Guttmann on the occasion of his 70th birthday.

  17. Vitellogenin synthesis in primary cultures of fish liver cells as endpoint for in vitro screening of the (anti)estrogenic activity of chemical substances.

    Science.gov (United States)

    Navas, José M; Segner, Helmut

    2006-10-25

    Concern over possible adverse effects of endocrine-disrupting compounds on fish has caused the development of appropriate testing methods. In vitro screening assays may provide initial information on endocrine activities of a test compound and thereby may direct and optimize subsequent testing. Induction of vitellogenin (VTG) is used as a biomarker of exposure of fish to estrogen-active substances. Since VTG induction can be measured not only in vivo but also in fish hepatocytes in vitro, the use of VTG induction response in isolated fish liver cells has been suggested as in vitro screen for identifying estrogenic-active substances. The main advantages of the hepatocyte VTG assay are considered its ability to detect effects of estrogenic metabolites, since hepatocytes in vitro remain metabolically competent, and its ability to detect both estrogenic and anti-estrogenic effects. In this article, we critically review the current knowledge on the VTG response of cultured fish hepatocytes to (anti)estrogenic substances. In particular, we discuss the sensitivity, specificity, and variability of the VTG hepatocyte assay. In addition, we review the available data on culture factors influencing basal and induced VTG production, the response to natural and synthetic estrogens as well as to xenoestrogens, the detection of indirect estrogens, and the sources of assay variability. The VTG induction in cultured fish hepatocytes is clearly influenced by culture conditions (medium composition, temperature, etc.) and culture system (hepatocyte monolayers, aggregates, liver slices, etc.). The currently available database on estrogen-mediated VTG induction in cultured teleost hepatocytes is too small to support conclusive statements on whether there exist systematic differences of the VTG response between in vitro culture systems, VTG analytical methods or fish species. The VTG hepatocyte assay detects sensitively natural and synthetic estrogens, whereas the response to

  18. Buccal cell FISH and blood PCR-Y detect high rates of X chromosomal mosaicism and Y chromosomal derivatives in patients with Turner syndrome.

    Science.gov (United States)

    Freriks, Kim; Timmers, Henri J L M; Netea-Maier, Romana T; Beerendonk, Catharina C M; Otten, Barto J; van Alfen-van der Velden, Janiëlle A E M; Traas, Maaike A F; Mieloo, Hanneke; van de Zande, Guillaume W H J F L; Hoefsloot, Lies H; Hermus, Ad R M M; Smeets, Dominique F C M

    2013-09-01

    Turner syndrome (TS) is the result of (partial) X chromosome monosomy. In general, the diagnosis is based on karyotyping of 30 blood lymphocytes. This technique, however, does not rule out tissue mosaicism or low grade mosaicism in the blood. Because of the associated risk of gonadoblastoma, mosaicism is especially important in case this involves a Y chromosome. We investigated different approaches to improve the detection of mosaicisms in 162 adult women with TS (mean age 29.9 ± 10.3). Standard karyotyping identified 75 patients (46.3%) with a non-mosaic monosomy 45,X. Of these 75 patients, 63 underwent additional investigations including FISH on buccal cells with X- and Y-specific probes and PCR-Y on blood. FISH analysis of buccal cells revealed a mosaicism in 19 of the 63 patients (30.2%). In five patients the additional cell lines contained a (derivative) Y chromosome. With sensitive real-time PCR we confirmed the presence of this Y chromosome in blood in three of the five cases. Although Y chromosome material was established in ovarian tissue in two patients, no gonadoblastoma was found. Our results confirm the notion that TS patients with 45,X on conventional karyotyping often have tissue specific mosaicisms, some of which include a Y chromosome. Although further investigations are needed to estimate the risk of gonadoblastoma in patients with Y chromosome material in buccal cells, we conclude that FISH or real-time PCR on buccal cells should be considered in TS patients with 45,X on standard karyotyping.

  19. 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.

  20. 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.

  1. 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)

  2. Comet-FISH with rDNA probes for the analysis of mutagen-induced DNA damage in plant cells.

    Science.gov (United States)

    Kwasniewska, Jolanta; Grabowska, Marta; Kwasniewski, Miroslaw; Kolano, Bozena

    2012-06-01

    We used comet-fluorescence in situ hybridization (FISH) in the model plant species Crepis capillaris following exposure of seedlings to maleic hydrazide (MH). FISH with 5S and 25S rDNA probes was applied to comets obtained under alkaline conditions to establish whether these DNA regions were preferentially involved in comet tail formation. MH treatment induced significant fragmentation of nuclear DNA and of rDNA loci. A 24-h post-treatment recovery period allowed a partial reversibility of MH-induced damage on nuclear and rDNA regions. Analyses of FISH signals demonstrated that rDNA sequences were always involved in tail formation and that 5S rDNA was more frequently present in the tail than 25S rDNA, regardless of treatment. The involvement of 25S rDNA in nucleolus formation and differences in chromatin structure between the two loci may explain the different susceptibility of the 25S and 5S rDNA regions to migrate into the tail. This work is the first report on the application of FISH to comet preparations from plants to analyze the distribution and repair of DNA damage within specific genomic regions after mutagenic treatment. Moreover, our work suggests that comet-FISH in plants may be a useful tool for environmental monitoring assessment.

  3. Alabama ESI: FISH (Fish Polygons)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data set contains sensitive biological resource data for marine, estuarine, and freshwater fish species in Alabama. Vector polygons in this data set represent...

  4. Virginia ESI: FISH (Fish Polygons)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data set contains sensitive biological resource data for marine, estuarine, anadromous, and brackishwater fish species in Virginia. Vector polygons in this data...

  5. Maryland ESI: FISH (Fish Polygons)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data set contains sensitive biological resource data for marine, estuarine, anadromous, and freshwater fish species in Maryland. Vector polygons in this data...

  6. Hawaii ESI: FISH (Fish Polygons)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data set contains sensitive biological resource data for reef, marine, estuarine, and native stream fish species in coastal Hawaii. Vector polygons in this data...

  7. Louisiana ESI: FISH (Fish Polygons)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data set contains sensitive biological resource data for freshwater (inland) fish species in coastal Louisiana. Vector polygons represent water-bodies and other...

  8. Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression

    Directory of Open Access Journals (Sweden)

    Huarong Guo

    2012-09-01

    Full Text Available p21CIP1/WAF1 is a p53-target gene in response to cellular DNA damage. Here we report the development of a fish cell biosensor system for high throughput genotoxicity detection of new drugs, by stably integrating two reporter plasmids of pGL3-p21-luc (human p21 promoter linked to firefly luciferase and pRL-CMV-luc (CMV promoter linked to Renilla luciferase into marine flatfish flounder gill (FG cells, referred to as p21FGLuc. Initial validation of this genotoxicity biosensor system showed that p21FGLuc cells had a wild-type p53 signaling pathway and responded positively to the challenge of both directly acting genotoxic agents (bleomycin and mitomycin C and indirectly acting genotoxic agents (cyclophosphamide with metabolic activation, but negatively to cyclophosphamide without metabolic activation and the non-genotoxic agents ethanol and D-mannitol, thus confirming a high specificity and sensitivity, fast and stable response to genotoxic agents for this easily maintained fish cell biosensor system. This system was especially useful in the genotoxicity detection of Di(2-ethylhexyl phthalate (DEHP, a rodent carcinogen, but negatively reported in most non-mammalian in vitro mutation assays, by providing a strong indication of genotoxicity for DEHP. A limitation for this biosensor system was that it might give false positive results in response to sodium butyrate and any other agents, which can trans-activate the p21 gene in a p53-independent manner.

  9. Comparison of IHC, FISH and RT-PCR methods for detection of ALK rearrangements in 312 non-small cell lung cancer patients in Taiwan.

    Directory of Open Access Journals (Sweden)

    Yi-Cheng Wu

    Full Text Available BACKGROUND: Recently Echinoderm microtubule-associated protein-like 4- anaplastic lymphoma kinase (EML4-ALK fusion gene has become an important biomarker for ALK tyrosine kinase inhibitor (crizotinib treatment in NSCLC. However, the best detection method and the significance of EML4-ALK variant types remain uncertain. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR, fluorescence in Situ hybridization (FISH and Immunohistochemical (IHC stain were performed on tumor tissues of 312 NSCLC patients for detection of ALK rearrangements. Mutation analyses for EGFR and KRAS genes were also performed. RESULTS: Thirteen of the 312 patients (4.17% had ALK rearrangements detected by RT-PCR. If RT-PCR data was used as the gold standard, FISH tests had a low sensitivity (58.33%, but very good specificity (99.32%. IHC stain had better sensitivity (91.67% than FISH, but lower specificity (79.52%, when the cut off was IHC2+. All of the 8 patients with high abundance of EML4-ALK positive cells in tumor tissues (assessed by the signal intensities of the RT-PCR product, were also have high expression of ALK protein (IHC3+, and positive for FISH, except one failed in FISH. Variants 3a+3b (4/5, 80% of EML4-ALK fusion gene were more common to have high abundance of EML4-ALK positive cells in tumor tissues than variant 1 (1/3, 33.3%. Meta-analysis of the published data of 2273 NSCLC patients revealed that variant 3 (23/44, 52.3% was the most common type in Chinese population, while variant 1 (28/37, 75.7% was most common in Caucasian. CONCLUSIONS: Among the three detection methods, RT-PCR could detect not only the presence of EML4-ALK fusion gene and their variant types, but also the abundance of EML4-ALK positive cells in NSCLC tumor tissues. The latter two factors might affect the treatment response to anti-ALK inhibitor. Including RT-PCR as a diagnostic test for ALK inhibitor treatment in the prospective clinical trials is recommended.

  10. 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.

  11. 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...

  12. 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.

  13. Gonadal development in fish.

    Science.gov (United States)

    Nishimura, Toshiya; Tanaka, Minoru

    2014-01-01

    Vertebrate reproduction depends on the function of 2 distinct gametes, sperm and eggs, which develop in 2 different organs, the testis and the ovary. Testes and ovaries are composed of germ cells, supporting cells and interstitial cells. In this review, we describe the origin and the fate of these cell lineages and how they interact with each other to form sexually dimorphic reproductive organs in medaka. We delineate how the temporally different association and establishment of these lineages contribute to a variety of seemingly different sex differentiation processes among teleost fish. Thus, teleosts represent an intriguing group in which to study the fundamental processes of gonadal development through comparing conserved and unique mechanisms.

  14. Biomonitoring Study of Heavy Metals in Blood from a Cement Factory Based Community

    Directory of Open Access Journals (Sweden)

    Bank M.S.

    2014-07-01

    Full Text Available Little is known about the effects of cement factory pollution, emissions, and kiln dust on contaminant exposure in human populations, including school environments, in close proximity to these point sources. In Ravena, New York, USA and vicinity, environmental pollution from a local cement plant is considered significant and substantial according to the United States Environmental Protection Agency’s Toxic Release Inventory, published in 2006, 2007, and 2010. We hypothesized that cement factory based communities, such as the one in Ravena, NY, may be differentially exposed to heavy metals, including mercury, via dust, soil, and air in addition to any contributions from fish consumption, dental amalgams, smoking habits, and occupational exposures, etc. Here we report measurements of several heavy metals in blood (Pb, Cd, As, Hg, Se and Al and, for comparative purposes, total mercury in hair from a local (six-mile radius population of Caucasian adults and children. We also report and synthesize local atmospheric emissions inventory information and new indoor air data (NYSERDA, 2011 from the local school which is situated directly across the street (within 750 feet from the cement factory and quarry. In addition, to our human and environmental heavy metal results we also discuss scientific outreach coordination, and public health action opportunities that will likely have wide applicability for other community and environmental health studies confronting similar pollution issues.

  15. 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…

  16. Fugu double U6 promoter-driven long double-stranded RNA inhibits proliferation of viral hemorrhagic septicemia virus (VHSV) in fish cell lines.

    Science.gov (United States)

    Kim, Min Sun; Jee, Bo Young; Cho, Mi Young; Kim, Jin Woo; Jeong, Hyun Do; Kim, Ki Hong

    2012-06-01

    A long double-stranded RNA (dsRNA)-producing vector driven by fugu double U6 promotors, in which the two promoters were arranged in a head-to-head fashion, was newly constructed. To determine whether the DNA-vector-based long dsRNAs can induce sequence-specific RNA interference (RNAi), Epithelioma papulosum cyprini (EPC) cells and chinook salmon embryonic (CHSE-214) cells were transfected with the long dsRNA vector targeting the G gene of VHSV, and its effect on expression of the G gene and viral proliferation was investigated. The sequence-specific inhibitory effect was further confirmed by analysis of interferon (IFN)-triggered Mx1 gene expression and cross-protection against infectious hematopoietic necrosis virus (IHNV). The fugu double U6 promoter-driven vector successfully produced long dsRNAs in EPC cells, a system that allows continuous production of long dsRNAs in transfected cells. The plasmid-based long dsRNAs targeting the VHSV G gene effectively suppressed G gene expression, but control dsRNAs targeting the EGFP gene did not. Furthermore, there was no significant difference in Mx gene expression between cells transfected with the long dsRNA-producing vector and those transfected with the control empty vector. These results suggest that G gene expression was suppressed not by type-I-IFN-mediated nonspecific inhibition but in a sequence-specific manner. Both EPC and CHSE-214 cells transfected with plasmids producing long dsRNAs targeting the VHSV G gene were protected against VHSV infection but were not protected against IHNV infection, suggesting sequence-specific RNAi-mediated inhibition of viral proliferation. In conclusion, we show, for the first time, long-dsRNA-mediated RNAi in fish cells. The DNA-vector-based long dsRNAs may provide an efficient tool for analysis of gene function in fish cells without preliminary burdensome work for selection of effective siRNA clones, and it may be applied as an antiviral measure in cultured fish.

  17. Apolipoprotein E genotype status affects habitual human blood mononuclear cell gene expression and its response to fish oil intervention

    NARCIS (Netherlands)

    Matualatupauw, J.C.; Radonjic, M.; Rest, van de O.; Groot, de C.P.G.M.; Geleijnse, J.M.; Müller, M.R.; Afman, L.A.

    2016-01-01

    Scope
    People who carry the apolipoprotein E4 (APOE4) single nucleotide polymorphism have an increased risk of cardiovascular disease (CVD). Fish-oil supplementation may help in the prevention of CVD, though interindividual differences in the response to n-3 PUFAs have been observed. We aimed to

  18. STIFFNESS CHANGES OF THE CUPULA ASSOCIATED WITH THE MECHANICS OF HAIR-CELLS IN THE FISH LATERAL-LINE

    NARCIS (Netherlands)

    van Netten, S.M.; KHANNA, SM

    1994-01-01

    Cupular vibration in the lateral-line canal of fish was measured in response to motion of the fluid in the canal by laser-heterodyne interferometry. The results show that the mechanical output/input ratio of the cupula depends on the stimulus amplitude; the cupula thus behaves nonlinearly. The nonli

  19. Globin X is a six-coordinate globin that reduces nitrite to nitric oxide in fish red blood cells

    Science.gov (United States)

    Corti, Paola; Xue, Jianmin; Tejero, Jesús; Wajih, Nadeem; Sun, Ming; Stolz, Donna B.; Tsang, Michael; Kim-Shapiro, Daniel B.; Gladwin, Mark T.

    2016-01-01

    The discovery of novel globins in diverse organisms has stimulated intense interest in their evolved function, beyond oxygen binding. Globin X (GbX) is a protein found in fish, amphibians, and reptiles that diverged from a common ancestor of mammalian hemoglobins and myoglobins. Like mammalian neuroglobin, GbX was first designated as a neuronal globin in fish and exhibits six-coordinate heme geometry, suggesting a role in intracellular electron transfer reactions rather than oxygen binding. Here, we report that GbX to our knowledge is the first six-coordinate globin and the first globin protein apart from hemoglobin, found in vertebrate RBCs. GbX is present in fish erythrocytes and exhibits a nitrite reduction rate up to 200-fold faster than human hemoglobin and up to 50-fold higher than neuroglobin or cytoglobin. Deoxygenated GbX reduces nitrite to form nitric oxide (NO) and potently inhibits platelet activation in vitro, to a greater extent than hemoglobin. Fish RBCs also reduce nitrite to NO and inhibit platelet activation to a greater extent than human RBCs, whereas GbX knockdown inhibits this nitrite-dependent NO signaling. The description of a novel, six-coordinate globin in RBCs with dominant electron transfer and nitrite reduction functionality provides new insights into the evolved signaling properties of ancestral heme-globins. PMID:27407144

  20. Apolipoprotein E genotype status affects habitual human blood mononuclear cell gene expression and its response to fish oil intervention

    NARCIS (Netherlands)

    Matualatupauw, J.C.; Radonjic, M.; Rest, O. van de; Groot, L.C.P.G.M. de; Geleijnse, J.M.; Müller, M.; Afman, L.A.

    2016-01-01

    Scope: People who carry the apolipoprotein E4 (APOE4) single nucleotide polymorphism have an increased risk of cardiovascular disease (CVD). Fish-oil supplementation may help in the prevention of CVD, though interindividual differences in the response to n-3 PUFAs have been observed. We aimed to ass

  1. 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.

  2. 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.

  3. 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.

  4. Fishing Access Areas

    Data.gov (United States)

    Vermont Center for Geographic Information — The Vermont Fish & Wildlife Department maintains developed fishing access areas. These sites provide public access to waters in Vermont for shore fishing...

  5. Calcium response of KCl-excited populations of ventricular myocytes from the European sea bass (Dicentrarchus labrax): a promising approach to integrate cell-to-cell heterogeneity in studying the cellular basis of fish cardiac performance.

    Science.gov (United States)

    Ollivier, Hélène; Marchant, James; Le Bayon, Nicolas; Servili, Arianna; Claireaux, Guy

    2015-10-01

    Climate change challenges the capacity of fishes to thrive in their habitat. However, through phenotypic diversity, they demonstrate remarkable resilience to deteriorating conditions. In fish populations, inter-individual variation in a number of fitness-determining physiological traits, including cardiac performance, is classically observed. Information about the cellular bases of inter-individual variability in cardiac performance is scarce including the possible contribution of excitation-contraction (EC) coupling. This study aimed at providing insight into EC coupling-related Ca(2+) response and thermal plasticity in the European sea bass (Dicentrarchus labrax). A cell population approach was used to lay the methodological basis for identifying the cellular determinants of cardiac performance. Fish were acclimated at 12 and 22 °C and changes in intracellular calcium concentration ([Ca(2+)]i) following KCl stimulation were measured using Fura-2, at 12 or 22 °C-test. The increase in [Ca(2+)]i resulted primarily from extracellular Ca(2+) entry but sarcoplasmic reticulum stores were also shown to be involved. As previously reported in sea bass, a modest effect of adrenaline was observed. Moreover, although the response appeared relatively insensitive to an acute temperature change, a difference in Ca(2+) response was observed between 12- and 22 °C-acclimated fish. In particular, a greater increase in [Ca(2+)]i at a high level of adrenaline was observed in 22 °C-acclimated fish that may be related to an improved efficiency of adrenaline under these conditions. In conclusion, this method allows a rapid screening of cellular characteristics. It represents a promising tool to identify the cellular determinants of inter-individual variability in fishes' capacity for environmental adaptation.

  6. 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.

  7. 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.

  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.

    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.

  9. 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.

  10. 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...

  11. Activation of Pax7-positive cells in a non-contractile tissue contributes to regeneration of myogenic tissues in the electric fish S. macrurus.

    Directory of Open Access Journals (Sweden)

    Christopher M Weber

    Full Text Available The ability to regenerate tissues is shared across many metazoan taxa, yet the type and extent to which multiple cellular mechanisms come into play can differ across species. For example, urodele amphibians can completely regenerate all lost tissues, including skeletal muscles after limb amputation. This remarkable ability of urodeles to restore entire limbs has been largely linked to a dedifferentiation-dependent mechanism of regeneration. However, whether cell dedifferentiation is the fundamental factor that triggers a robust regeneration capacity, and whether the loss or inhibition of this process explains the limited regeneration potential in other vertebrates is not known. Here, we studied the cellular mechanisms underlying the repetitive regeneration of myogenic tissues in the electric fish S. macrurus. Our in vivo microinjection studies of high molecular weight cell lineage tracers into single identified adult myogenic cells (muscle or noncontractile muscle-derived electrocytes revealed no fragmentation or cellularization proximal to the amputation plane. In contrast, ultrastructural and immunolabeling studies verified the presence of myogenic stem cells that express the satellite cell marker Pax7 in mature muscle fibers and electrocytes of S. macrurus. These data provide the first example of Pax-7 positive muscle stem cells localized within a non-contractile electrogenic tissue. Moreover, upon amputation, Pax-7 positive cells underwent a robust replication and were detected exclusively in regions that give rise to myogenic cells and dorsal spinal cord components revealing a regeneration process in S. macrurus that is dependent on the activation of myogenic stem cells for the renewal of both skeletal muscle and the muscle-derived electric organ. These data are consistent with the emergent concept in vertebrate regeneration that different tissues provide a distinct progenitor cell population to the regeneration blastema, and these

  12. Activation of Pax7-positive cells in a non-contractile tissue contributes to regeneration of myogenic tissues in the electric fish S. macrurus.

    Science.gov (United States)

    Weber, Christopher M; Martindale, Mark Q; Tapscott, Stephen J; Unguez, Graciela A

    2012-01-01

    The ability to regenerate tissues is shared across many metazoan taxa, yet the type and extent to which multiple cellular mechanisms come into play can differ across species. For example, urodele amphibians can completely regenerate all lost tissues, including skeletal muscles after limb amputation. This remarkable ability of urodeles to restore entire limbs has been largely linked to a dedifferentiation-dependent mechanism of regeneration. However, whether cell dedifferentiation is the fundamental factor that triggers a robust regeneration capacity, and whether the loss or inhibition of this process explains the limited regeneration potential in other vertebrates is not known. Here, we studied the cellular mechanisms underlying the repetitive regeneration of myogenic tissues in the electric fish S. macrurus. Our in vivo microinjection studies of high molecular weight cell lineage tracers into single identified adult myogenic cells (muscle or noncontractile muscle-derived electrocytes) revealed no fragmentation or cellularization proximal to the amputation plane. In contrast, ultrastructural and immunolabeling studies verified the presence of myogenic stem cells that express the satellite cell marker Pax7 in mature muscle fibers and electrocytes of S. macrurus. These data provide the first example of Pax-7 positive muscle stem cells localized within a non-contractile electrogenic tissue. Moreover, upon amputation, Pax-7 positive cells underwent a robust replication and were detected exclusively in regions that give rise to myogenic cells and dorsal spinal cord components revealing a regeneration process in S. macrurus that is dependent on the activation of myogenic stem cells for the renewal of both skeletal muscle and the muscle-derived electric organ. These data are consistent with the emergent concept in vertebrate regeneration that different tissues provide a distinct progenitor cell population to the regeneration blastema, and these progenitor cells

  13. In vitro assessment of DNA damage after short- and long-term exposure to benzo(a)pyrene using RAPD and the RTG-2 fish cell line.

    Science.gov (United States)

    Castaño, Argelia; Becerril, Concepción

    2004-08-18

    Genotoxins present in the aquatic environment are often associated with the decline or disappearance of many wild populations. The hazard assessment of chemicals requires sensitive and specific tests to study the genotoxic effects in order to establish the maximum allowable chemical concentrations prior to the release to the environment. We have previously shown that an established fish cell line (RTG-2) together with the random amplified polymorphic DNA (RAPD) technique, can be used to detect alterations in the DNA caused by direct acting genotoxins. The current study takes this a step further and examines in the same system the effect of a pro-mutagen benzo(a)pyrene (B(a)P) at different concentrations (0.05, 0.1, and 0.5 microg/ml) and at different exposure periods (1, 2, 3, 15, and 30 days). After comparing DNA fingerprints from control and exposed cells, both qualitative and quantitative analysis show an increase in the instability in the DNA fingerprint of exposed cells over a time- and concentration-dependent manner. At the higher concentration (0.5 microg/ml) three out the four primers showed altered bands after 1 day of exposure, while after 3 days all used primers showed an altered pattern. At the lower concentration of B(a)P (0.05 microg/ml) the appearance of new bands was observed with a 100% level of reproducibility after 30 days of exposure suggesting an inheritance of the altered DNA. We conclude that this in vitro system is useful to evaluate genotoxic effects, both after acute and chronic exposures and of direct and non-direct acting genotoxins. Cultured cells can be considered as genetically homogenous populations. Therefore, in vitro systems permits us to undertake mechanistic studies avoiding the interference of polymorphisms inherent in the in vivo systems. Furthermore, the RTG-2 fish cell line combined with a RAPD assay could be used in studies of hazard identification in risk assessment protocols of chemicals.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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...

  19. 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.

  20. Abortive infection of snakehead fish vesiculovirus in ZF4 cells was associated with the RLRs pathway activation by viral replicative intermediates.

    Science.gov (United States)

    Wang, Wenwen; Asim, Muhammad; Yi, Lizhu; Hegazy, Abeer M; Hu, Xianqin; Zhou, Yang; Ai, Taoshan; Lin, Li

    2015-03-18

    Snakehead fish vesiculovirus (SHVV) is a negative strand RNA virus which can cause great economic losses in fish culture. To facilitate the study of SHVV-host interactions, the susceptibility of zebrafish embryonic fibroblast cell line (ZF4) to the SHVV was investigated in this report. The results showed that high amount of viral mRNAs and cRNAs were detected at the 3 h post-infection. However, the expressions of the viral mRNAs and cRNA were decreased dramatically after 6 h post-infection. In addition, the expressions of interferon (IFN) and interferon-induced GTP-binding protein Mx were all up regulated significantly at the late stage of the infection. Meanwhile, the expressions of Retinoic acid-inducible gene I (RIG-I) and Melanoma differentiation-associated gene 5 (MDA5) were also all up-regulated significantly during the infection. Two isoforms of DrLGP2 from zebrafish were also cloned and analyzed. Interestingly, the expression of DrLGP2a but not DrLGP2b was significantly up-regulated at both mRNA and protein levels, indicating that the two DrLGP2 isoforms might play different roles during the SHVV infection. Transfection experiment showed that viral replicative intermediates were required for the activation of IFN-α expression. Taken together, the abortive infection of SHVV in ZF4 cells was associated with the activation of RLRs pathway, which was activated by viral replicative intermediates.

  1. Low concentrations of caffeine induce asymmetric cell division as observed in vitro by means of the CBMN-assay and iFISH.

    Science.gov (United States)

    Hatzi, Vasiliki I; Karakosta, Maria; Barszczewska, Katarzyna; Karachristou, Ioanna; Pantelias, Gabriel; Terzoudi, Georgia I

    2015-11-01

    The dual role of caffeine as a chromosomal damage inducer and G2/M-checkpoint abrogator is well known but it is observed mainly at relatively high concentrations. At low concentrations, caffeine enhances the cytogenetic effects of several carcinogens and its intake during pregnancy has been recently reported to cause adverse birth outcomes. Interestingly, a threshold below which this association is not apparent was not identified. Since chromosomal abnormalities and aneuploidy are the major genetic etiologies of spontaneous abortions and adverse birth outcomes, we re-evaluate here the effects of caffeine at the cytogenetic level and propose a model for the mechanisms involved. Our hypothesis is that low caffeine concentrations affect DNA replication and cause chromosomal aberrations and asymmetric cell divisions not easily detected at metaphase since damaged cells are delayed during their G2/M-phase transition and the low caffeine concentrations cannot abrogate the G2-checkpoint. To test this hypothesis, caffeine-induced chromatid breaks and micronuclei in peripheral blood lymphocytes (PBLs) were evaluated in vitro after low caffeine concentration exposures, followed by a short treatment with 4mM of caffeine to abrogate the G2-checkpoint. The results show a statistically significant increase in chromatid breaks at caffeine concentrations ≥1mM. When caffeine was applied for G2/M-checkpoint abrogation, a statistically significant increase in chromatid breaks, compared to an active checkpoint, was only observed at 4mM of caffeine. The potential of low concentrations to induce asymmetric cell divisions was tested by applying a methodology combining the cytochalasin-B mediated cytokinesis-block micronucleus assay (CBMN) with interphase FISH (iFISH), using selected centromeric probes. Interestingly, low caffeine concentrations induce a dose dependent aneuploidy through asymmetric cell divisions, which are caused by misalignment of chromosomes through a mechanism

  2. Ecology of Listeria spp. in a fish farm and molecular typing of Listeria monocytogenes from fish farming and processing companies.

    Science.gov (United States)

    Miettinen, Hanna; Wirtanen, Gun

    2006-11-01

    This study focused on the ecology of Listeria monocytogenes in a fish farm by following the changes in its occurrence in different types of samples for a three year period. In addition, L. monocytogenes isolates from different seafood industry areas were compared with pulsed field gel electrophoresis (PFGE) typing to discover possible associations between primary production, further processing and final products. Weather conditions were found to have a strong influence on the probability of finding Listeria spp. in a fish farm environment. The number of samples contaminated with Listeria spp. was typically bigger after rainy periods. Brook and river waters as well as other runoff waters seemed to be the main contamination source at the farm studied. The farmed fish originally found to carry L. monocytogenes become gradually Listeria free. The time needed for the purification of the fish was several months. The sea bottom soil samples were the ones that preserved the L. monocytogenes contamination the longest time. It can be stated that the fish and fish farm equipment studied did not spread listeria contamination. On the contrary, they were found to suffer from listeria contamination coming from outside sources like the brook water. There was a wide range of different L. monocytogenes PFGE-pulsotypes (30) found at 15 Finnish fish farms and fish processing factories. L. monocytogenes isolates from the final products often belonged to the same pulsotypes as did the isolates from the processing environment as well as from the raw fish. This suggests that, in addition to the fish processing factory environment, the fish raw materials are important sources of L. monocytogenes contamination in final products.

  3. The Fishing Cat

    Institute of Scientific and Technical Information of China (English)

    孙雅飞; 乐伟国

    2008-01-01

    @@ 一、故事内容 A cat goes fishing every day. He wants to eat fish, but he can't catch any fish. One day, he goes to the river as usual. Suddenly, a fish comes out. He catches the fish and putsthe fish in the basket. He's very happy, but he forgest to put the lid on the basket.

  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. Breaking the host range: mandarin fish is susceptible to a vesiculovirus derived from snakehead fish.

    Science.gov (United States)

    Liu, Xiaodan; Wen, Yi; Hu, Xianqin; Wang, Wenwen; Liang, Xufang; Li, Jun; Vakharia, Vikram; Lin, Li

    2015-04-01

    Members of the genus Vesiculovirus, which belongs to the family Rhabdoviridae, can cause great economic loss in fish culture. In the present report, a vesiculovirus [named snakehead fish vesiculovirus (SHVV)] was isolated from diseased hybrid snakehead fish. SHVV shared 94 % nucleotide sequence identity at the genomic level with Siniperca chuatsi rhabdovirus (SCRV), which infects mandarin fish (S. chuatsi). We showed that SHVV was able to replicate and proliferate well in SSN-1 cells, which originate from striped snakehead fish (Channa striatus). Furthermore, mandarin fish was susceptible to SHVV by bath exposure, as well as by intraperitoneal injection. The infected fish showed typical clinical signs of rhabdovirus infection, including haemorrhage and oedema. Histopathological analysis revealed that extensive inflammation and necrosis were observed in the spleen, kidney, liver, heart and brain of the moribund mandarin fish. These results will shed new light on the epidemic of vesiculovirus infections among fish.

  11. 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...

  12. Fish gelatin.

    Science.gov (United States)

    Boran, Gokhan; Regenstein, Joe M

    2010-01-01

    Gelatin is a multifunctional ingredient used in foods, pharmaceuticals, cosmetics, and photographic films as a gelling agent, stabilizer, thickener, emulsifier, and film former. As a thermoreversible hydrocolloid with a narrower gap between its melting and gelling temperatures, both of which are below human body temperature, gelatin provides unique advantages over carbohydrate-based gelling agents. Gelatin is mostly produced from pig skin, and cattle hides and bones. Some alternative raw materials have recently gained attention from both researchers and the industry not just because they overcome religious concerns shared by Jews and Muslims but also because they provide, in some cases, technological advantages over mammalian gelatins. Fish skins from a number of fish species are among the other sources that have been comprehensively studied as sources for gelatin production. Fish skins have a significant potential for the production of high-quality gelatin with different melting and gelling temperatures over a much wider range than mammalian gelatins, yet still have a sufficiently high gel strength and viscosity. Gelatin quality is industrially determined by gel strength, viscosity, melting or gelling temperatures, the water content, and microbiological safety. For gelatin manufacturers, yield from a particular raw material is also important. Recent experimental studies have shown that these quality parameters vary greatly depending on the biochemical characteristics of the raw materials, the manufacturing processes applied, and the experimental settings used for quality control tests. In this review, the gelatin quality achieved from different fish species is reviewed along with the experimental procedures used to determine gelatin quality. In addition, the chemical structure of collagen and gelatin, the collagen-gelatin conversion, the gelation process, and the gelatin market are discussed.

  13. 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.

  14. Fish hemoglobins

    Directory of Open Access Journals (Sweden)

    P.C. de Souza

    2007-06-01

    Full Text Available Vertebrate hemoglobin, contained in erythrocytes, is a globular protein with a quaternary structure composed of 4 globin chains (2 alpha and 2 beta and a prosthetic group named heme bound to each one. Having myoglobin as an ancestor, hemoglobin acquired the capacity to respond to chemical stimuli that modulate its function according to tissue requirements for oxygen. Fish are generally submitted to spatial and temporal O2 variations and have developed anatomical, physiological and biochemical strategies to adapt to the changing environmental gas availability. Structurally, most fish hemoglobins are tetrameric; however, those from some species such as lamprey and hagfish dissociate, being monomeric when oxygenated and oligomeric when deoxygenated. Fish blood frequently possesses several hemoglobins; the primary origin of this finding lies in the polymorphism that occurs in the globin loci, an aspect that may occasionally confer advantages to its carriers or even be a harmless evolutionary remnant. On the other hand, the functional properties exhibit different behaviors, ranging from a total absence of responses to allosteric regulation to drastic ones, such as the Root effect.

  15. 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.

  16. Efficient display of active Geotrichum sp. lipase on Pichia pastoris cell wall and its application as a whole-cell biocatalyst to enrich EPA and DHA in fish oil.

    Science.gov (United States)

    Pan, Xiao-Xing; Xu, Li; Zhang, Yan; Xiao, Xiao; Wang, Xiao-Feng; Liu, Yun; Zhang, Hou-jin; Yan, Yun-Jun

    2012-09-26

    Geotrichum sp. lipase (GSL) was first displayed on the cell wall of Pichia pastoris on the basis of the a-agglutinin anchor system developed in Saccharomyces cerevisiae . Surface display levels were monitored using Western blotting, immunofluorescence miscroscopy, and fluorescence-activated cell sorting analysis. Lipase activity of the yeast whole cells reached a maximum at 273 ± 2.4 U/g of dry cells toward olive oil after 96 h of culture at 30 °C, with optimal pH and temperature at 7.5 and 45 °C, respectively. Displayed GSL exhibited relatively high stability between pH 6.0 and 8.0 and retained >70% of the maximum activity. The surface-displayed lipase retained 80% of its original activity after incubation at 45 °C for 4 h. Moreover, the GSL-displaying yeast whole cells were then used as a biocatalyst to enrich eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish oil on the basis of selective hydrolysis. As a result, EPA and DHA increased from 1.53 and 24.1% in the original fish oil to 1.85 and 30.86%, which were increases of 1.21- and 1.29-fold, respectively. The total yield of EPA and DHA reached 46.62%.

  17. DNA damage in the kidney tissue cells of the fish Rhamdia quelen after trophic contamination with aluminum sulfate

    Directory of Open Access Journals (Sweden)

    Tatiane Klingelfus

    2015-01-01

    Full Text Available Abstract Even though aluminum is the third most common element present in the earth's crust, information regarding its toxicity remains scarce. It is known that in certain cases, aluminum is neurotoxic, but its effect in other tissues is unknown. The aim of this work was to analyze the genotoxic potential of aluminum sulfate in kidney tissue of the fish Rhamdia quelen after trophic contamination for 60 days. Sixty four fish were subdivided into the following groups: negative control, 5 mg, 50 mg and 500 mg of aluminum sulfate per kg of fish. Samples of the posterior kidney were taken and prepared to obtain mitotic metaphase, as well as the comet assay. The three types of chromosomal abnormalities (CA found were categorized as chromatid breaks, decondensation of telomeric region, and early separation of sister chromatids. The tests for CA showed that the 5 mg/kg and 50 mg/kg doses of aluminum sulfate had genotoxic potential. Under these treatments, early separation of the sister chromatids was observed more frequently and decondensation of the telomeric region tended to increase in frequency. We suggest that structural changes in the proteins involved in DNA compaction may have led to the decondensation of the telomeric region, making the DNA susceptible to breaks. Moreover, early separation of the sister chromatids may have occurred due to changes in the mobility of chromosomes or proteins that keep the sister chromatids together. The comet assay confirmed the genotoxicity of aluminum sulfate in the kidney tissue of Rhamdia quelen at the three doses of exposure.

  18. 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.

  19. Genome-wide identification of mononuclear cell DNA methylation sites potentially affected by fish oil supplementation in young infants

    DEFF Research Database (Denmark)

    Lind, Mads Vendelbo; Martino, D; Harsløf, Laurine Bente Schram;

    2015-01-01

    Recent evidence suggests that the effects of n-3LCPUFA might be mediated through epigenetic mechanisms, especially DNA-methylation, during pregnancy and early life. A randomized trial was conducted in 133 9-mo-old, infants who received 3.8g/day of fish oil (FO) or sunflower oil (SO) for 9 mo....... In a subset of 12 children, buffy-coat DNA was extracted before and after intervention and analyzed on Illumina-Human-Methylation 450-arrays to explore genome-wide differences between the FO and SO groups. Genome-wide-methylation analysis did not reveal significant differences between groups after adjustment...

  20. 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.

  1. 斑马鱼视网膜细胞图像的分割研究%Image segmentation of zebra fish retinal cells

    Institute of Scientific and Technical Information of China (English)

    杜培明; 汪宁宁; 史晓丽

    2012-01-01

    针对斑马鱼视网膜细胞图像分割时细胞粘连比较严重的情况,采用了边界距离变换和模拟浸水原理的分水岭变换相结合的方法。首先采用距离变换的方法提取细胞区域的局部极小值作为种子点,合并距离过小的错误种子点,然后使用分水岭算法进行分割。实验结果表明,该方法能够有效地分割斑马鱼视网膜细胞图像,成功地解决了分水岭变换中粘连细胞的过分割问题,且计算速度快。%For image segmentation cases of zebra fish retinal cells in serious overlapped, the paper uses the boundary distance transform and the principle of immersion simulation method of combining watershed transform. The basic principle is extracting the local minimum values of ceils as a regional seeds point by using distance transformation. It deletes the error seed points which are too close, and then divided by the watershed algorithm. Experimental results show that the method can effectively split the image of zebra fish retinal cells. The method has successfully solved the over-segmentation problem of watershed transformation in cells adhe- sion, and the calculation is fast.

  2. Effects of a silver nanomaterial on cellular organelles and time course of oxidative stress in a fish cell line (PLHC-1).

    Science.gov (United States)

    Bermejo-Nogales, A; Fernández, M; Fernández-Cruz, M L; Navas, J M

    2016-12-01

    Among the nanomaterials currently in commercial products, those based on silver are the most used, and so there is a high probability that silver nanoparticles (AgNPs) will be released into aquatic environments where they could adversely affect aquatic organisms, including fish. Taking this into account, the aim of the present work was to characterize in depth the mechanisms underlying the toxic action of AgNPs using fish cell lines, determining specifically the contribution of alterations in cellular structures and oxidative stress time course to the cytotoxicity of AgNPs. Since liver plays a key role in detoxification, the hepatoma cell line PLHC-1 was used. Exposure to AgNPs (NM-300K, obtained from the Joint Research Centre Repository) caused alterations at the lysosomal and mitochondrial levels at lower concentrations than those that disrupted plasma membrane (evaluated by means of neutral red, alamarBlue, and 5-carboxyfluorescein diacetate, acetoxymethyl ester assays respectively). AgNO3, used as a control Ag(+) ion source, produced similar cytotoxic effects but at lower concentrations than AgNPs. Both silver forms caused oxidative disruption but the initial response was delayed in AgNPs until 6h of exposure. Transmission electron microscopy analysis also evidenced the disruption of mitochondrial structures in cells exposed to cytotoxic concentrations of both forms of silver. At non-cytotoxic concentrations, AgNPs were detected inside the nucleoli and mitochondria, thereby pointing to long-term effects. The present work evidences the mutual interaction between the induction of oxidative stress and the alterations of cellular structures, particularly mitochondria, as cytotoxicity mechanisms not exclusively associated to NPs.

  3. Exclusive multipotency and preferential asymmetric divisions in post-embryonic neural stem cells of the fish retina.

    Science.gov (United States)

    Centanin, Lázaro; Ander, Janina-J; Hoeckendorf, Burkhard; Lust, Katharina; Kellner, Tanja; Kraemer, Isabel; Urbany, Cedric; Hasel, Eva; Harris, William A; Simons, Benjamin D; Wittbrodt, Joachim

    2014-09-01

    The potency of post-embryonic stem cells can only be addressed in the living organism, by labeling single cells after embryonic development and following their descendants. Recently, transplantation experiments involving permanently labeled cells revealed multipotent neural stem cells (NSCs) of embryonic origin in the medaka retina. To analyze whether NSC potency is affected by developmental progression, as reported for the mammalian brain, we developed an inducible toolkit for clonal labeling and non-invasive fate tracking. We used this toolkit to address post-embryonic stem cells in different tissues and to functionally differentiate transient progenitor cells from permanent, bona fide stem cells in the retina. Using temporally controlled clonal induction, we showed that post-embryonic retinal NSCs are exclusively multipotent and give rise to the complete spectrum of cell types in the neural retina. Intriguingly, and in contrast to any other vertebrate stem cell system described so far, long-term analysis of clones indicates a preferential mode of asymmetric cell division. Moreover, following the behavior of clones before and after external stimuli, such as injuries, shows that NSCs in the retina maintained the preference for asymmetric cell division during regenerative responses. We present a comprehensive analysis of individual post-embryonic NSCs in their physiological environment and establish the teleost retina as an ideal model for studying adult stem cell biology at single cell resolution.

  4. Effect of fish oil on lateral mobility of prostaglandin F2α (FP) receptors and spatial distribution of lipid microdomains in bovine luteal cell plasma membrane in vitro.

    Science.gov (United States)

    Plewes, M R; Burns, P D; Graham, P E; Hyslop, R M; Barisas, B G

    2017-01-01

    Lipid microdomains are ordered regions on the plasma membrane of cells, rich in cholesterol and sphingolipids, ranging in size from 10 to 200 nm in diameter. These lipid-ordered domains may serve as platforms to facilitate colocalization of intracellular signaling proteins during agonist-induced signal transduction. It is hypothesized that fish oil will disrupt the lipid microdomains, increasing spatial distribution of these lipid-ordered domains and lateral mobility of the prostaglandin (PG) F2α (FP) receptors in bovine luteal cells. The objectives of this study were to examine the effects of fish oil on (1) the spatial distribution of lipid microdomains, (2) lateral mobility of FP receptors, and (3) lateral mobility of FP receptors in the presence of PGF2α on the plasma membrane of bovine luteal cells in vitro. Bovine ovaries were obtained from a local abattoir and corpora lutea were digested using collagenase. In experiment 1, lipid microdomains were labeled using cholera toxin subunit B Alexa Fluor 555. Domains were detected as distinct patches on the plasma membrane of mixed luteal cells. Fish oil treatment decreased fluorescent intensity in a dose-dependent manner (P oil treatment on lateral mobility of FP receptors. Fish oil treatment increased microdiffusion and macrodiffusion coefficients of FP receptors as compared to control cells (P oil-treated cells (P oil treatment. Lateral mobility of receptors was decreased within 5 min following the addition of ligand for control cells (P oil-treated cells (P > 0.10). The data presented provide strong evidence that fish oil causes a disruption in lipid microdomains and affects lateral mobility of FP receptors in the absence and presence of PGF2α.

  5. 77 FR 44593 - Burlington Coat Factory Warehouse Corporation, Provisional Acceptance of a Settlement Agreement...

    Science.gov (United States)

    2012-07-30

    ... COMMISSION Burlington Coat Factory Warehouse Corporation, Provisional Acceptance of a Settlement Agreement... accepted Settlement Agreement with Burlington Coat Factory Warehouse Corporation, containing a civil... Factory Warehouse Corporation CPSC Docket No. 12-C0008 Settlement Agreement 1. In accordance with...

  6. Exclusive Multipotency and Preferential Asymmetric Divisions in Post-Embryonic Neural Stem Cells of the Fish Retina

    OpenAIRE

    Centanin, L.; Ander, J.-J.; Hoeckendorf, B.; Lust, K.; T. Kellner; Kraemer, I.; Urbany, C.; Hasel, E.; Harris, W.A.; Simons, B. D.; Wittbrodt, J

    2014-01-01

    This is the final version of the article. It has been published by The Company of Biologists Ltd in Development here: http://dev.biologists.org/content/early/2014/08/19/dev.109892.long. The potency of post-embryonic stem cells can only be addressed in the living organism, by labeling single cells after embryonic development and following their descendants. Recently, transplantation experiments involving permanently labeled cells revealed multipotent neural stem cells (NSCs) of embryonic or...

  7. The Gamma Factory proposal for CERN

    CERN Document Server

    Krasny, Mieczyslaw Witold

    2015-01-01

    This year, 2015, marks the centenary of the publication of Einsteins Theory of General Relativity and it has been named the International Year of Light and light-based technologies by the UN General Assembly. It is thus timely to discuss the possibility of broadening the present CERN research program by including a new component based on a novel concept of the light source which could pave a way towards a multipurpose Gamma Factory. The proposed light source could be realized at CERN by using the infrastructure of the existing accelerators. It could push the intensity limits of the presently operating light-sources by at least 7 orders of magnitude, reaching the flux of the order of 10^17 photons/s, in the particularly interesting gamma-ray energy domain of 1 < Ephoton < 400 MeV. This domain is out of reach for the FEL-based light sources. The energy-tuned, quasi-monochromatic gamma beams, together with the gamma-beam-driven, high intensity secondary beams of polarized positrons, polarized muons, neutro...

  8. Forecasting Sales in a Sugar Factory

    Directory of Open Access Journals (Sweden)

    Vassilios ASSIMAKOPOULOS

    2011-12-01

    Full Text Available Beets’ cultivation and sugar production represent one of the most important parts of Greek agricultural economy. A careful and well-organized planning of the production as well as the determination of an accurate safety stock is important for sugar industry, as for many other companies and organizations, in order to define the production quantity which leads to maximum revenues and profits. Forecasting, and especially widely used statistical forecasting techniques, is the best way for policymakers to organize their activities and company’s production and make the appropriate adjustments. Apparently, management information systems and forecasting support packages play a leading role in this area, since the amount of data under process is usually quite large and demands an automated procedure to effectively produce and evaluate forecasts. In this case study, “Pythia”, an expert forecasting platform developed by the Forecasting and Strategy Unit of the National Technical University of Athens, was implemented on a monthly data series regarding sugar sales of a Greek sugar factory for the years 2000-2005, bringing theory and practice together. Additionally, the methods or combinations of methods which are well suited for this time series are highlighted based on three error indices. Finally, the results of the study and conclusions are considered and perspectives of progress and development in the field of forecasting are contemplated.

  9. New Physics at a Super Flavor Factory

    CERN Document Server

    Browder, Thomas E; Pirjol, Dan; Soni, Amarjit; Zupan, Jure

    2009-01-01

    The potential of a Super Flavor Factory (SFF) for searches of New Physics is reviewed. While very high luminosity B physics is assumed to be at the core of the program, its scope for extensive charm and tau studies are also emphasized. The possibility to run at the Upsilon(5S) as well as at the Upsilon(4S) is also very briefly discussed; in principle, this could provide very clean measurements of B_s decays. The strength and reach of a SFF is most notably due to the possibility of examining an impressive array of very clean observables. The angles and the sides of the unitarity triangle can be determined with unprecedented accuracy. These serve as a reference for New Physics (NP) sensitive decays such as B^+ ->tau^+ nu and penguin dominated hadronic decay modes, providing tests of generic NP scenarios with an accuracy of a few percent. Besides, very precise studies of direct and time dependent CP asymmetries in radiative B decays and forward-backward asymmetry studies in B -> X_s l^+ l^- and numerous null tes...

  10. Future Accelerators, Muon Colliders, and Neutrino Factories

    Energy Technology Data Exchange (ETDEWEB)

    Richard A Carrigan, Jr.

    2001-12-19

    Particle physics is driven by five great topics. Neutrino oscillations and masses are now at the fore. The standard model with extensions to supersymmetry and a Higgs to generate mass explains much of the field. The origins of CP violation are not understood. The possibility of extra dimensions has raised tantalizing new questions. A fifth topic lurking in the background is the possibility of something totally different. Many of the questions raised by these topics require powerful new accelerators. It is not an overstatement to say that for some of the issues, the accelerator is almost the experiment. Indeed some of the questions require machines beyond our present capability. As this volume attests, there are parts of the particle physics program that have been significantly advanced without the use of accelerators such as the subject of neutrino oscillations and many aspects of the particle-cosmology interface. At this stage in the development of physics, both approaches are needed and important. This chapter first reviews the status of the great accelerator facilities now in operation or coming on within the decade. Next, midrange possibilities are discussed including linear colliders with the adjunct possibility of gamma-gamma colliders, muon colliders, with precursor neutrino factories, and very large hadron colliders. Finally visionary possibilities are considered including plasma and laser accelerators.

  11. Fish Tales

    Energy Technology Data Exchange (ETDEWEB)

    McLerran, L.

    2010-07-06

    This talk is about fishing and the friendships that have resulted in its pursuit. It is also about theoretical physics, and the relationship of imagination and fantasy to the establishment of ideas about nature. Fishermen, like theoretical physicists, are well known for their inventive imaginations. Perhaps neither are as clever as sailors, who conceived of the mermaid. If one doubts the power of this fantasy, one should remember the ghosts of the many sailors who drowned pursuing these young nymphs. An extraordinary painting by J. Waterhouse is shown as Fig. 1. The enchantment of a mermaid must reflect an extraordinary excess of imagination on the part of the sailor, perhaps together with an impractical turn of mind. A consummated relationship with a mermaid is after all, by its very nature a fantasy incapable of realization. To a theoretical physicist, she is symbolic of many ideas we develop. There are many truths known to fisherman in which one might also find parallels to the goals of scientists: (1) A fish is the only animal that keeps growing after its death; (2) Nothing makes a fish bigger than almost being caught; (3) ''...of all the liars among mankind, the fisherman is the most trustworthy.'' (William Sherwood Fox, in Silken Lines and Silver Hooks); and (4) Men and fish are alike. They both get into trouble when they open their mouths. These quotes may be interpreted as reflecting skepticism regarding the honesty of fisherman, and probably do not reflect adequate admiration for a creative imagination. Is it fair to criticize a person for believing a falsehood that he or she sincerely believes to be true? The fisherman simultaneously invents the lie, and believes in it himself. The parallel with theoretical physics is perhaps only approximate, although we physicists may invent stories that we come to believe, on some rare occasions our ideas actually correspond to a more or less true descriptions of nature. These minor philosophical

  12. Identification of Insulin-Like Growth Factor-I Receptor (IGF-IR Gene Promoter-Binding Proteins in Estrogen Receptor (ER-Positive and ER-Depleted Breast Cancer Cells

    Directory of Open Access Journals (Sweden)

    Rive Sarfstein

    2010-03-01

    Full Text Available The insulin-like growth factor I receptor (IGF-IR has been implicated in the etiology of breast cancer. Overexpression of the IGF-IR gene is a typical feature of most primary breast cancers, whereas low IGF-IR levels are seen at advanced stages. Hence, evaluation of IGF-IR levels might be important for assessing prognosis. In the present study, we employed a proteomic approach based on DNA affinity chromatography followed either by mass spectroscopy (MS or Western blot analysis to identify transcription factors that may associate with the IGF-IR promoter in estrogen receptor (ER-positive and ER-depleted breast cancer cells. A biotinylated IGF-IR promoter fragment was bound to streptavidin magnetic beads and incubated with nuclear extracts of breast cancer cells. IGF-IR promoter-binding proteins were eluted with high salt and analyzed by MS and Western blots. Among the proteins that were found to bind to the IGF-IR promoter we identified zinc finger transcription factors Sp1 and KLF6, ER-, p53, c-jun, and poly (ADP-ribosylation polymerase. Furthermore, chromatin immune-precipitation (ChIP analysis confirmed the direct in vivo binding of some of these transcription factors to IGF-IR promoter DNA. The functional relevance of binding data was assessed by cotransfection experiments with specific expression vectors along with an IGF-IR promoter reporter. In summary, we identified nuclear proteins that are potentially responsible for the differential expression of the IGF-IR gene in ER-positive and ER-depleted breast cancer cells.

  13. Uniformity pattern and related criteria for two-level factorials

    Institute of Scientific and Technical Information of China (English)

    FANG; Kaitai; QIN; Hong

    2005-01-01

    In this paper,the study of projection properties of two-level factorials in view of geometry is reported.The concept of uniformity pattern is defined.Based on this new concept,criteria of uniformity resolution and minimum projection uniformity are proposed for comparing two-level factorials.Relationship between minimum projection uniformity and other criteria such as minimum aberration,generalized minimum aberration and orthogonality is made explict.This close relationship raises the hope of improving the connection between uniform design theory and factorial design theory.Our results provide a justification of orthogonality,minimum aberration,and generalized minimum aberration from a natural geometrical interpretation.

  14. Environmental radioactivity level at a mantle factory in eastern Zhejiang

    Institute of Scientific and Technical Information of China (English)

    YE Ji-Da; ZENG Guang-Jian; WU Zong-Mei; LIU Hong-Shi; NI Shi-Ying

    2003-01-01

    The γ radiation level, alpha potential energy of thoron and alpha surface contamination level at amantle factory in the east of Zhejiang Province are reported. The measured results show that the additional annual ef-fective dose equivalent absorbed by the worker in some workshops of the factory was higher than 15mSv-management limit. The alpha surface contamination on the workers' hands in some workshops was 10 timesmore than the management limit of National Standard (0.04 Bq/cm2). The mantle factory's main danger to body wasinternal irradiation of α rays from thoron aerosol.

  15. Deep Fish.

    Science.gov (United States)

    Ishaq, Omer; Sadanandan, Sajith Kecheril; Wählby, Carolina

    2017-01-01

    Zebrafish ( Danio rerio) is an important vertebrate model organism in biomedical research, especially suitable for morphological screening due to its transparent body during early development. Deep learning has emerged as a dominant paradigm for data analysis and found a number of applications in computer vision and image analysis. Here we demonstrate the potential of a deep learning approach for accurate high-throughput classification of whole-body zebrafish deformations in multifish microwell plates. Deep learning uses the raw image data as an input, without the need of expert knowledge for feature design or optimization of the segmentation parameters. We trained the deep learning classifier on as few as 84 images (before data augmentation) and achieved a classification accuracy of 92.8% on an unseen test data set that is comparable to the previous state of the art (95%) based on user-specified segmentation and deformation metrics. Ablation studies by digitally removing whole fish or parts of the fish from the images revealed that the classifier learned discriminative features from the image foreground, and we observed that the deformations of the head region, rather than the visually apparent bent tail, were more important for good classification performance.

  16. Abortive Infection of Snakehead Fish Vesiculovirus in ZF4 Cells Was Associated with the RLRs Pathway Activation by Viral Replicative Intermediates

    Directory of Open Access Journals (Sweden)

    Wenwen Wang

    2015-03-01

    Full Text Available Snakehead fish vesiculovirus (SHVV is a negative strand RNA virus which can cause great economic losses in fish culture. To facilitate the study of SHVV-host interactions, the susceptibility of zebrafish embryonic fibroblast cell line (ZF4 to the SHVV was investigated in this report. The results showed that high amount of viral mRNAs and cRNAs were detected at the 3 h post-infection. However, the expressions of the viral mRNAs and cRNA were decreased dramatically after 6 h post-infection. In addition, the expressions of interferon (IFN and interferon-induced GTP-binding protein Mx were all up regulated significantly at the late stage of the infection. Meanwhile, the expressions of Retinoic acid-inducible gene I (RIG-I and Melanoma differentiation-associated gene 5 (MDA5 were also all up-regulated significantly during the infection. Two isoforms of DrLGP2 from zebrafish were also cloned and analyzed. Interestingly, the expression of DrLGP2a but not DrLGP2b was significantly up-regulated at both mRNA and protein levels, indicating that the two DrLGP2 isoforms might play different roles during the SHVV infection. Transfection experiment showed that viral replicative intermediates were required for the activation of IFN-α expression. Taken together, the abortive infection of SHVV in ZF4 cells was associated with the activation of RLRs pathway, which was activated by viral replicative intermediates.

  17. True 3q chromosomal amplification in squamous cell lung carcinoma by FISH and aCGH molecular analysis: impact on targeted drugs.

    Directory of Open Access Journals (Sweden)

    Matteo Brunelli

    Full Text Available Squamous lung carcinoma lacks specific "ad hoc" therapies. Amplification of chromosome 3q is the most common genomic aberration and this region harbours genes having role as novel targets for therapeutics. There is no standard definition on how to score and report 3q amplification. False versus true 3q chromosomal amplification in squamous cell lung carcinoma may have tremendous impact on trials involving drugs which target DNA zones mapping on 3q. Forty squamous lung carcinomas were analyzed by FISH to assess chromosome 3q amplification. aCGH was performed as gold-standard to avoid false positive amplifications. Three clustered patterns of fluorescent signals were observed. Eight cases out of 40 (20% showed ≥8 3q signals. Twenty out of 40 (50% showed from 3 to 7 signals. The remaining showed two fluorescent signals (30%. When corrected by whole chromosome 3 signals, only cases with ≥8 signals maintained a LSI 3q/CEP3 ratio >2. Only the cases showing 3q amplification by aCGH (+3q25.3-3q27.3 showed ≥8 fluorescent signals at FISH evidencing a 3q/3 ratio >2. The remaining cases showed flat genomic portrait at aCGH on chromosome 3. We concluded that: 1 absolute copy number of 3q chromosomal region may harbour false positive interpretation of 3q amplification in squamous cell carcinoma; 2 a case results truly "amplified for chromosome 3q" when showing ≥8 fluorescent 3q signals; 3 trials involving drugs targeting loci on chromosome 3q in squamous lung carcinoma therapy have to consider false versus true 3q chromosomal amplification.

  18. Interphase FISH Demonstrates that Human Adipose Stromal Cells Maintain a High Level of Genomic Stability in Long-Term Culture

    OpenAIRE

    2008-01-01

    Human adipose stromal cells (ASCs) reside within the stromal-vascular fraction (SVF) in fat tissue, can be readily isolated, and include stem-like cells that may be useful for therapy. An important consideration for clinical application and functional studies of stem/progenitor cells is their capacity to maintain chromosome stability in culture. In this study, cultured ASC populations and ASC clones were evaluated at intervals for maintenance of chromosome stability. Uncultured SVF (uSVF) cel...

  19. Make your company a talent factory.

    Science.gov (United States)

    Ready, Douglas A; Conger, Jay A

    2007-06-01

    Despite the great sums of money companies dedicate to talent management systems, many still struggle to fill key positions - limiting their potential for growth in the process. Virtually all the human resource executives in the authors' 2005 survey of 40 companies around the world said that their pipeline of high-potential employees was insufficient to fill strategic management roles. The survey revealed two primary reasons for this. First, the formal procedures for identifying and developing next-generation leaders have fallen out of sync with what companies need to grow or expand into new markets. To save money, for example, some firms have eliminated positions that would expose high-potential employees to a broad range of problems, thus sacrificing future development opportunities that would far outweigh any initial savings from the job cuts. Second, HR executives often have trouble keeping top leaders' attention on talent issues, despite those leaders' vigorous assertions that obtaining and keeping the best people is a major priority. If passion for that objective doesn't start at the top and infuse the culture, say the authors, talent management can easily deteriorate into the management of bureaucratic routines. Yet there are companies that can face the future with confidence. These firms don't just manage talent, they build talent factories. The authors describe the experiences of two such corporations - consumer products icon Procter & Gamble and financial services giant HSBC Group -that figured out how to develop and retain key employees and fill positions quickly to meet evolving business needs. Though each company approached talent management from a different direction, they both maintained a twin focus on functionality (rigorous talent processes that support strategic and cultural objectives) and vitality (management's emotional commitment, which is reflected in daily actions).

  20. Black hole as a wormhole factory

    Science.gov (United States)

    Kim, Sung-Won; Park, Mu-In

    2015-12-01

    There have been lots of debates about the final fate of an evaporating black hole and the singularity hidden by an event horizon in quantum gravity. However, on general grounds, one may argue that a black hole stops radiation at the Planck mass (ħc / G) 1 / 2 ∼10-5 g, where the radiated energy is comparable to the black hole's mass. And also, it has been argued that there would be a wormhole-like structure, known as "spacetime foam", due to large fluctuations below the Planck length (ħG /c3) 1 / 2 ∼10-33 cm. In this paper, as an explicit example, we consider an exact classical solution which represents nicely those two properties in a recently proposed quantum gravity model based on different scaling dimensions between space and time coordinates. The solution, called "Black Wormhole", consists of two different states, depending on its mass parameter M and an IR parameter ω: For the black hole state (with ωM2 > 1 / 2), a non-traversable wormhole occupies the interior region of the black hole around the singularity at the origin, whereas for the wormhole state (with ωM2 Censorship" by the existence of a wormhole-like structure which protects the naked singularity even after the black hole evaporation. One could understand the would-be wormhole inside the black hole horizon as the result of microscopic wormholes created by "negative" energy quanta which have entered the black hole horizon in Hawking radiation process; the quantum black hole could be a wormhole factory! It is found that this speculative picture may be consistent with the recent " ER = EPR " proposal for resolving the black hole entanglement debates.

  1. Overview of the nearby supernova factory

    Energy Technology Data Exchange (ETDEWEB)

    Aldering, Greg; Adam, Gilles; Antilogus, Pierre; Astier, Pierre; Bacon, Roland; Bongard, S.; Bonnaud, C.; Copin, Yannick; Hardin, D.; Howell, D. Andy; Lemmonnier, Jean-Pierre; Levy, J.-M.; Loken, S.; Nugent, Peter; Pain, Reynald; Pecontal, Arlette; Pecontal, Emmanuel; Perlmutter, Saul; Quimby, Robert; Schahmaneche, Kyan; Smadja, Gerard; Wood-Vasey, W. Michael

    2002-07-29

    The Nearby Supernova Factory (SNfactory) is an international experiment designed to lay the foundation for the next generation of cosmology experiments (such as CFHTLS, wP, SNAP and LSST) which will measure the expansion history of the Universe using Type Ia supernovae. The SNfactory will discover and obtain frequent lightcurve spectrophotometry covering 3200-10000 {angstrom} for roughly 300 Type Ia supernovae at the low-redshift end of the smooth Hubble flow. The quantity, quality, breadth of galactic environments, and homogeneous nature of the SNfactory dataset will make it the premier source of calibration for the Type Ia supernova width-brightness relation and the intrinsic supernova colors used for K-correction and correction for extinction by host-galaxy dust. This dataset will also allow an extensive investigation of additional parameters which possibly influence the quality of Type Ia supernovae as cosmological probes. The SNfactory search capabilities and follow-up instrumentation include wide-field CCD imagers on two 1.2-m telescopes (via collaboration with the Near Earth Asteroid Tracking team at JPL and the QUEST team at Yale), and a two-channel integral-field-unit optical spectrograph/imager being fabricated for the University of Hawaii 2.2-m telescope. In addition to ground-based follow-up, UV spectra for a subsample of these supernovae will be obtained with HST. The pipeline to obtain, transfer via wireless and standard internet, and automatically process the search images is in operation. Software and hardware development is now underway to enable the execution of follow-up spectroscopy of supernova candidates at the Hawaii 2.2-m telescope via automated remote control of the telescope and the IFU spectrograph/imager.

  2. The Experiment Factory: standardizing behavioral experiments

    Directory of Open Access Journals (Sweden)

    Vanessa V Sochat

    2016-04-01

    Full Text Available The administration of behavioral and experimental paradigms for psychology research is hindered by lack of a coordinated effort to develop and deploy standardized paradigms. While several frameworks (de Leeuw (2015; McDonnell et al. (2012; Mason and Suri (2011; Lange et al. (2015 have provided infrastructure and methods for individual research groups to develop paradigms, missing is a coordinated effort to develop paradigms linked with a system to easily deploy them. This disorganization leads to redundancy in development, divergent implementations of conceptually identical tasks, disorganized and error-prone code lacking documentation, and difficulty in replication. The ongoing reproducibility crisis in psychology and neuroscience research (Baker (2015; Open Science Collaboration (2015 highlights the urgency of this challenge: reproducible research in behavioral psychology is conditional on deployment of equivalent experiments. A large, accessible repository of experiments for researchers to develop collaboratively is most efficiently accomplished through an open source framework. Here we present the Experiment Factory, an open source framework for the development and deployment of web-based experiments. The modular infrastructure includes experiments, virtual machines for local or cloud deployment, and an application to drive these components and provide developers with functions and tools for further extension. We release this infrastructure with a deployment (http://www.expfactory.org that researchers are currently using to run a set of over 80 standardized web-based experiments on Amazon Mechanical Turk. By providing open source tools for both deployment and development, this novel infrastructure holds promise to bring reproducibility to the administration of experiments, and accelerate scientific progress by providing a shared community resource of psychological paradigms.

  3. Oscillations of factorial cumulants to factorial moments ratio from an eikonal approach

    Energy Technology Data Exchange (ETDEWEB)

    Beggio, P.C., E-mail: beggio@uenf.br [Universidade Federal do Rio Grande do Sul – UFRGS, Porto Alegre, RS (Brazil); Universidade Estadual do Norte Fluminense Darcy Ribeiro – UENF, Campos dos Goytacazes, RJ (Brazil)

    2013-09-02

    We study the factorial moments (F{sub q}), the factorial cumulants (K{sub q}) and the ratio of K{sub q} to F{sub q} (H{sub q}=K{sub q}/F{sub q}) in pp/pp{sup ¯} collisions using an updated approach, in which the multiplicity distribution is related to the eikonal function. The QCD inspired eikonal model adopted contains contributions of quark–quark, quark–gluon and gluon–gluon interactions. Our work shows that the approach can reproduce the collision energy dependence of the F{sub q} moments, correctly predicts that the first minimum of the H{sub q} lies around q=5 and qualitatively reproduces the oscillations of the H{sub q} moments, as shown in the experimental data and predicted by QCD at preasymptotic energy. The result of this study seems to indicate that the H{sub q} oscillations are manifestations of semihard component in the multiparticle production process. Predictions for multiplicity distribution and H{sub q} moments at the LHC energy of √(s)=14 TeV are presented.

  4. Utilization of Spirulina platensis for wastewater treatment in fermented rice noodle factory

    Directory of Open Access Journals (Sweden)

    Sumontip Bunnag

    2010-12-01

    Full Text Available Spirulina platensis is a single-cell blue-green alga which belongs to the family Oscillatoriaceae.S. platensis can grow in polluted water and it has been widely used to remove excess nitrate-nitrogenand total phosphorus content in wastewater from noodle factories. This study was aimed to investigatethe effect of wastewater on growth of S. platensis and to examine its potential to reduce nitrate-nitrogenand total phosphorus contents in wastewater from fermented rice noodle factory. Twenty five percentwastewatermixed Zarrouk’s medium gave rise to the highest growth rate of S. platensis. Optical density(OD of S. platensis culture increased from 0.07±0.04 to 0.24±0.12. Also, the results exhibited the leastvalue of nitrate-nitrogen and total phosphorus contents when performed in 25 % and 100 %wastewater-mixed Zarrouk’s media, respectively.

  5. Alginate Biosynthesis Factories in Pseudomonas fluorescens: Localization and Correlation with Alginate Production Level.

    Science.gov (United States)

    Maleki, Susan; Almaas, Eivind; Zotchev, Sergey; Valla, Svein; Ertesvåg, Helga

    2015-12-11

    Pseudomonas fluorescens is able to produce the medically and industrially important exopolysaccharide alginate. The proteins involved in alginate biosynthesis and secretion form a multiprotein complex spanning the inner and outer membranes. In the present study, we developed a method by which the porin AlgE was detected by immunogold labeling and transmission electron microscopy. Localization of the AlgE protein was found to depend on the presence of other proteins in the multiprotein complex. No correlation was found between the number of alginate factories and the alginate production level, nor were the numbers of these factories affected in an algC mutant that is unable to produce the precursor needed for alginate biosynthesis. Precursor availability and growth phase thus seem to be the main determinants for the alginate production rate in our strain. Clustering analysis demonstrated that the alginate multiprotein complexes were not distributed randomly over the entire outer cell membrane surface.

  6. Factorial kriging analysis applied to geological data from petroleum exploration

    Energy Technology Data Exchange (ETDEWEB)

    Jaquet, O.

    1989-10-01

    A regionalized variable, thickness of the reservoir layer, from a gas field is decomposed by factorial kriging analysis. Maps of the obtained components may be associated with depositional environments that are favorable for petroleum exploration.

  7. Lead levels of Culex mosquito larvae inhabiting lead utilizing factory

    Institute of Scientific and Technical Information of China (English)

    Kitvatanachai S; Apiwathnasorn C; Leemingsawat S; Wongwit W; Overgaard HJ

    2011-01-01

    Objective: To determine lead level primarily in Culex quinquefasciatus (Cx. quinquefasciatus), and Culex gelidus (Cx. gelidus) larvae inhabiting lead consuming factories, and to putatively estimate eco-toxicological impact of effluents from the firms. Methods: Third instars larvae were sampled by standard dipping method and lead concentrations in the larvae and their respective surrounding factory aquatic environments were determined through standard atomic absorption spectrophotometry (AAS). Results: Cx. quinquefasciatus was the most abundant species followed by Cx. gelidus. The levels of lead were higher in the Cx. quinquefasciatus (1.08-47.47 μg/g), than in the wastewaters surface (0.01-0.78 μg/mL) from the factories or closer areas around factories. Other species were not reaching the criteria for lead determination. Conclusions: The Cx.quinquefasciatus larvae can bio-accumulate the metal and can potentially serve as a biomarker of lead contamination, to complemente conventional techniques.

  8. Application of fiber laser for a Higgs factory

    Energy Technology Data Exchange (ETDEWEB)

    Chou, W.

    2014-06-04

    This paper proposes a medium size(~6km) circular Higgs factory based on a photon collider. The recent breakthrough in fiber laser technology by means of a coherent amplifier network makes such a collider feasible and probably also affordable.

  9. Factorial Schur functions via the six vertex model

    CERN Document Server

    McNamara, Peter J

    2009-01-01

    For a particular set of Boltzmann weights and a particular boundary condition for the six vertex model in statistical mechanics, we compute explicitly the partition function and show it to be equal to a factorial Schur function.

  10. D and Ds Hadronic Branching Fractions at B Factories

    Energy Technology Data Exchange (ETDEWEB)

    Pappagallo, Marco; /Bari U. /INFN, Bari

    2008-01-07

    Recent measurements of hadronic branching fractions of D and D{sub s} mesons, performed by the BABAR and Belle experiments at the asymmetric e{sup +}e{sup -} B factories colliders PEP II and KEKB, are reviewed.

  11. Effects of a diet high in fish oil (MaxEPA) on the formation of micronucleated erythrocytes in blood and on the number of atypical acinar cell foci induced in rat pancreas by Azaserine

    NARCIS (Netherlands)

    Appel, M.J.; Woutersen, R.A.

    2004-01-01

    The present study was performed to investigate the influence of fish oil on the genotoxic effects of azaserine, using the formation of micronucleated erythrocytes as a measure for the degree of initiating potency and the number and size of putative preneoplastic pancreatic atypical acinar cell foci

  12. Fishing amplifies forage fish population collapses.

    Science.gov (United States)

    Essington, Timothy E; Moriarty, Pamela E; Froehlich, Halley E; Hodgson, Emma E; Koehn, Laura E; Oken, Kiva L; Siple, Margaret C; Stawitz, Christine C

    2015-05-26

    Forage fish support the largest fisheries in the world but also play key roles in marine food webs by transferring energy from plankton to upper trophic-level predators, such as large fish, seabirds, and marine mammals. Fishing can, thereby, have far reaching consequences on marine food webs unless safeguards are in place to avoid depleting forage fish to dangerously low levels, where dependent predators are most vulnerable. However, disentangling the contributions of fishing vs. natural processes on population dynamics has been difficult because of the sensitivity of these stocks to environmental conditions. Here, we overcome this difficulty by collating population time series for forage fish populations that account for nearly two-thirds of global catch of forage fish to identify the fingerprint of fisheries on their population dynamics. Forage fish population collapses shared a set of common and unique characteristics: high fishing pressure for several years before collapse, a sharp drop in natural population productivity, and a lagged response to reduce fishing pressure. Lagged response to natural productivity declines can sharply amplify the magnitude of naturally occurring population fluctuations. Finally, we show that the magnitude and frequency of collapses are greater than expected from natural productivity characteristics and therefore, likely attributed to fishing. The durations of collapses, however, were not different from those expected based on natural productivity shifts. A risk-based management scheme that reduces fishing when populations become scarce would protect forage fish and their predators from collapse with little effect on long-term average catches.

  13. Sport Fishing Regulations

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — The regulations for sport fishing on St. Vincent National Wildlife Refuge are outlined in this document. Fishing is only permitted from sunrise to sunset, and only...

  14. Fish Springs pond snail

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Communication scenario between the branch of Listing and Recovery, Fish and Wildlife Enhancement, and Fish Springs National Wildlife Refuge (NWR), in regards to the...

  15. Fish tapeworm infection

    Science.gov (United States)

    Fish tapeworm infection is an intestinal infection with the tapeworm parasite found in fish. ... The fish tapeworm ( Diphyllobothrium latum ) is the largest parasite that infects humans. Humans become infected when they eat raw ...

  16. Got a Sick Fish?

    Science.gov (United States)

    ... Welfare Veterinary Careers Public Health Got a sick fish? Fish with disease can show a variety of signs. If you notice your pet fish having any unusual disease signs, contact your veterinarian ...

  17. Development of law relating to factories in India

    OpenAIRE

    M. K. Sahu

    2015-01-01

    The rapid growth of industrial town and factories has paved the way to develop our industrial legislation accordingly. The Government of India never expressed their interest in framing separate legislation vis-à-vis factories which resulted in implementation of the same statute which was enacted pre-independence. It was done by virtue of Art. 372 of the Constitution of India. However, the Constitutional Lawmakers created vacuum for the implementation of new statute in accordance with the dema...

  18. Noncommunicable disease risk profile of factory workers in Delhi

    Science.gov (United States)

    Kishore, Jugal; Kohli, Charu; Sharma, Pramod Kumar; Sharma, Ekta

    2012-01-01

    Background: Noncommunicable diseases (NCDs) are becoming more prevalent in India. The data for presence of NCDs and its risk factors among factory workers is deficient in India. Materials and Methods: A cross-sectional comparative study was carried out among 37 factory workers and equal number of comparable subjects from general population. Screening for presence of diabetes along with its risk factors was made in both the groups using pretested predesigned World Health Organization STEPwise approach to surveillance (WHO STEPS) questionnaire in rural area of Delhi. Data was analyzed using SPSS version 16 software. The estimation of risk in two groups was done with calculation of odds ratio (OR). P values less than 0.05 were considered significant. Results: A total of 74 participants were included in the present study. Hypertension and diabetes was present in 13.5 and 5.4% of factory workers and four (10.8%) and three (8.8%) subjects in comparative group, respectively. Seven (18.9%) factory and eight (21.6%) non-factory subjects fell in the category of current smoker or smokeless tobacco users. High density lipoprotein levels were found abnormal among one (2.7%) factory worker and nine (24.3%) subjects in comparative group (P-value = 0.01). Behavioral risk factors, alcohol consumption, and fruits and vegetable intake were significantly different among two groups. Conclusion: Factory workers were having better profile than non-factory subjects except for risk factors such as alcohol intake and inadequate fruits and vegetable intake. However, healthy worker effect phenomenon cannot be ruled out. PMID:23776324

  19. Noncommunicable disease risk profile of factory workers in Delhi

    Directory of Open Access Journals (Sweden)

    Jugal Kishore

    2012-01-01

    Full Text Available Background: Noncommunicable diseases (NCDs are becoming more prevalent in India. The data for presence of NCDs and its risk factors among factory workers is deficient in India. Materials and Methods: A cross-sectional comparative study was carried out among 37 factory workers and equal number of comparable subjects from general population. Screening for presence of diabetes along with its risk factors was made in both the groups using pretested predesigned World Health Organization STEPwise approach to surveillance (WHO STEPS questionnaire in rural area of Delhi. Data was analyzed using SPSS version 16 software. The estimation of risk in two groups was done with calculation of odds ratio (OR. P values less than 0.05 were considered significant. Results: A total of 74 participants were included in the present study. Hypertension and diabetes was present in 13.5 and 5.4% of factory workers and four (10.8% and three (8.8% subjects in comparative group, respectively. Seven (18.9% factory and eight (21.6% non-factory subjects fell in the category of current smoker or smokeless tobacco users. High density lipoprotein levels were found abnormal among one (2.7% factory worker and nine (24.3% subjects in comparative group (P-value = 0.01. Behavioral risk factors, alcohol consumption, and fruits and vegetable intake were significantly different among two groups. Conclusion: Factory workers were having better profile than non-factory subjects except for risk factors such as alcohol intake and inadequate fruits and vegetable intake. However, healthy worker effect phenomenon cannot be ruled out.

  20. Invariabilidad de la estructura factorial del "California Personality Inventory" (CPI

    Directory of Open Access Journals (Sweden)

    Horacio J. A. Rimoldi

    1976-01-01

    Full Text Available Tbe factor analysis of the California Personality Inventory (CPI and Rokeaeh Dogmatism Scale was performed using the results obtained from 161 college students, of both sexes, between 18 and 22 years of age, Four factors were extracted using the principal axes solution and graphic rotations. The results were compared with other factorial studíes, They showed a highly satistory agreement with some other solution strenghtening the factorial invariance of the CPI.

  1. Expert Meeting Report: Advanced Envelope Research for Factory Built Housing

    Energy Technology Data Exchange (ETDEWEB)

    Levy, E.; Mullens, M.; Tompos, E.; Kessler, B.; Rath, P.

    2012-04-01

    This report provides information about the expert meeting on advanced envelope research for factory built housing, hosted by the ARIES Collaborative on October 11, 2011, in Phoenix, Arizona. The goals of this meeting were to provide a comprehensive solution to the use of three previously selected advanced alternatives for factory-built wall construction, assess each option focusing on major issues relating to viability and commercial potential, and determine additional steps are required to reach this potential.

  2. Expert Meeting Report: Advanced Envelope Research for Factory Built Housing

    Energy Technology Data Exchange (ETDEWEB)

    Levy, E.; Mullens, M.; Tompos, E.; Kessler, B.; Rath, P.

    2012-04-01

    This report provides information about the Building America expert meeting on advanced envelope research for factory built housing, hosted by the ARIES Collaborative on October 11, 2011, in Phoenix, Arizona. The goals of this meeting were to provide a comprehensive solution to the use of three previously selected advanced alternatives for factory-built wall construction, assess each option focusing on major issues relating to viability and commercial potential, and determine additional steps are required to reach this potential.

  3. Economic and Environmental Sustainability of Factory Farming in the US

    OpenAIRE

    Nilsson, Asbjørn Lupo; Giersing, Josephine; Magrane, David; Breitenstein, Marcus

    2014-01-01

    This paper is written with the purpose of looking into sustainable development. More specifically, it will look at the relationship between the environmental and economical pillars of sustainability. In order for sustainable development to take place, the economy must be growing while maintaining earth’s natural resources. Factory farming might be strong from an economic point of view, but it does not seem to be environmentally friendly. Therefore we used factory farming as an example of an i...

  4. Black hole as a wormhole factory

    Directory of Open Access Journals (Sweden)

    Sung-Won Kim

    2015-12-01

    Full Text Available There have been lots of debates about the final fate of an evaporating black hole and the singularity hidden by an event horizon in quantum gravity. However, on general grounds, one may argue that a black hole stops radiation at the Planck mass (ħc/G1/2∼10−5 g, where the radiated energy is comparable to the black hole's mass. And also, it has been argued that there would be a wormhole-like structure, known as “spacetime foam”, due to large fluctuations below the Planck length (ħG/c31/2∼10−33 cm. In this paper, as an explicit example, we consider an exact classical solution which represents nicely those two properties in a recently proposed quantum gravity model based on different scaling dimensions between space and time coordinates. The solution, called “Black Wormhole”, consists of two different states, depending on its mass parameter M and an IR parameter ω: For the black hole state (with ωM2>1/2, a non-traversable wormhole occupies the interior region of the black hole around the singularity at the origin, whereas for the wormhole state (with ωM2<1/2, the interior wormhole is exposed to an outside observer as the black hole horizon is disappearing from evaporation. The black hole state becomes thermodynamically stable as it approaches the merging point where the interior wormhole throat and the black hole horizon merges, and the Hawking temperature vanishes at the exact merge point (with ωM2=1/2. This solution suggests the “Generalized Cosmic Censorship” by the existence of a wormhole-like structure which protects the naked singularity even after the black hole evaporation. One could understand the would-be wormhole inside the black hole horizon as the result of microscopic wormholes created by “negative” energy quanta which have entered the black hole horizon in Hawking radiation process; the quantum black hole could be a wormhole factory! It is found that this speculative picture may be consistent with the

  5. No Fishing Now,More Fish Later

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    Fishing ban for ecological purposes starts on the Pearl River Since April1,a two-month ban on fishing has been imposed on the Pearl River valley in south China.It is the first fishing ban in this area with the purpose of preserving biodiversity in China’s third longest

  6. Microbiological spoilage of fish and fish products

    DEFF Research Database (Denmark)

    Gram, Lone; Huss, Hans Henrik

    1996-01-01

    Spoilage of fresh and lightly preserved fish products is caused by microbial action. This paper reviews the current knowledge in terms of the microbiology of fish and fish products with particular emphasis on identification of specific spoilage bacteria and the qualitative and quantitative...

  7. Characterization of gonadotropic cells during continuous and seasonal spermatogenesis of two freshwater fish species: a histochemical and immunohistochemical study.

    Science.gov (United States)

    Nóbrega, Rafael Henrique; de Jesus, Lázaro Wender Oliveira; Honji, Renato Massaaki; Borella, Maria Inês

    2017-02-01

    This work describes gonadotropic (GtH) cells and their morphological and immunohistochemical changes during the spermatogenic cycle of Serrasalmus maculatus (continuous spermatogenesis) and Pimelodus maculatus (seasonal spermatogenesis). GtH cells, widely distributed in the proximal pars distalis of the adenohypophysis, were characterized as round-shaped cells with eccentric nucleus, and cytoplasm with basophilic secretory granules and a variable number of vacuoles for both species. Immunohistochemistry against β-follicle-stimulating hormone (Fsh) and β-luteinizing hormone (Lh) in adjacent sections showed two separated GtH-producing cell populations, and a third population where both GtHs are expressed in the same cell for both species. In the seasonal spermatogenesis of P. maculatus, GtH cells seemed to be more abundant during developing and spawning capable phases. In contrast, no cyclic changes were detected in the continuous spermatogenesis of S. maculatus, except for the strong immunoreaction for Fsh and Lh in males with intense spermiogenesis. We conclude that changes reported here might reflect the type of spermatogenic cycle (seasonal or continuous) which are under different regulatory mechanisms (environmental and internal cues) controlling the reproduction in these species.

  8. Single-cell gel electrophoresis assay in the ten spotted live-bearer fish, Cnesterodon decemmaculatus (Jenyns, 1842), as bioassay for agrochemical-induced genotoxicity.

    Science.gov (United States)

    Vera-Candioti, Josefina; Soloneski, Sonia; Larramendy, Marcelo L

    2013-12-01

    The ability of two 48 percent chlorpyrifos-based insecticides (Lorsban* 48E® and CPF Zamba®), two 50 percent pirimicarb-based insecticides (Aficida® and Patton Flow®), and two 48 percent glyphosate-based herbicides (Panzer® and Credit®) to induce DNA single-strand breaks in peripheral blood erythrocytes of Cnesterodon decemmaculatus (Jenyns, 1842) (Pisces, Poeciliidae) exposed under laboratory conditions was evaluated by the single-cell gel electrophoresis (SCGE) assay. In those fish exposed to Lorsban* 48E®, CPF Zamba®, Aficida®, Patton Flow®, Credit®, and Panzer®, a significant increase of the genetic damage was observed for all formulations regardless of the harvesting time. This genotoxic effect was achieved by an enhancement of Type II-IV comets and a concomitant decrease of Type 0-I comets over control values. A regression analysis revealed that the damage varied as a negative function of the exposure time in those Lorsban* 48E®- and Aficida®-treated fish. On the other hand, a positive correlation between damage increase and exposure time was achieved after Patton Flow® and Credit® treatment. Finally, no correlation was observed between increase in the genetic damage and exposure time after treatment with CPF Zamba® or Panzer®. These results highlight that all agrochemicals inflict primary genotoxic damage at the DNA level at sublethal concentrations, regardless of the exposure time of the aquatic organisms under study, at least within a period of 96 h of treatment.

  9. Picornaviruses and reoviruses of fishes

    Science.gov (United States)

    Winton, J.R.; Ahne, Winfried; Kurstak, E.

    1989-01-01

    The number of fish viruses isolated in cell culture or observed by electron microscopy continues to increase rapidly. Until recently, most viruses that were isolated from finfish and characterized were found to be members of the Rhabdoviridae, Iridoviridae, or Herpesviridae (Wolf and Mann 1980). In a comprehensive review of fish viruses published in 1984, there were no picornaviruses and only two reoviruses listed (Wolf 1984). The expansion of aquaculture into the rearing of new species at high density in different geographic areas, and the use of improved methods of detection that include newly developed cell lines and increased sampling effort, have led to the discovery of fish viruses representing nearly all families of animal viruses. Among the newest additions, are a member of the family Picornaviridae and several new viruses that belong within the Reoviridae.

  10. Changes in adipocyte cell size, gene expression of lipid metabolism markers, and lipolytic responses induced by dietary fish oil replacement in gilthead sea bream (Sparus aurata L.).

    Science.gov (United States)

    Cruz-Garcia, Lourdes; Sánchez-Gurmaches, Joan; Bouraoui, Lamia; Saera-Vila, Alfonso; Pérez-Sánchez, Jaume; Gutiérrez, Joaquim; Navarro, Isabel

    2011-04-01

    The effects of fish oil (FO) substitution by 66% vegetable oils in a diet with already 75% vegetable protein (66VO) on adipose tissue lipid metabolism of gilthead sea bream were analysed after a 14-month feeding trial. In the last 3 months of the experiment, a FO diet was administrated to a 66VO group (group 66VO/FO) as a finishing diet. Hormone-sensitive lipase (HSL) activity was measured in adipose tissue and adipocyte size, and HSL, lipoprotein lipase and liver X receptor gene expression in isolated adipocytes, on which lipolysis and glucose uptake experiments were also performed. Lipolysis was measured after incubation with tumour necrosis factor-α (TNFα), linoleic acid, and two conjugated linoleic acid isomers. Glucose uptake was analysed after TNFα or insulin administration. Our results show that FO replacement increased lipolytic activity and adipocyte cell size. The higher proportion of large cells observed in the 66VO group could be involved in their observed lower response to fatty acid treatments and lower insulin sensitivity. The 66VO/FO group showed a moderate return to the FO conditions. Therefore, FO replacement can affect the morphology and metabolism of gilthead sea bream adipocytes which could potentially affect other organs such as the liver.

  11. A protocol for adult somatic cell nuclear transfer in medaka fish (Oryzias latipes) with a high rate of viable clone formation.

    Science.gov (United States)

    Bubenshchikova, Ekaterina; Kaftanovskaya, Elena; Adachi, Tomoko; Hashimoto, Hisashi; Kinoshita, Masato; Wakamatsu, Yuko

    2013-12-01

    Previously, we successfully generated fully grown, cloned medaka (the Japanese rice fish, Oryzias latipes) using donor nuclei from primary culture cells of adult caudal fin tissue and nonenucleated recipient eggs that were heat shock-treated to induce diploidization of the nuclei. However, the mechanism of clone formation using this method is unknown, and the rate of adult clone formation is not high enough for studies in basic and applied sciences. To gain insight into the mechanism and increase the success rate of this method of clone formation, we tested two distinct nuclear transfer protocols. In one protocol, the timing of transfer of donor nuclei was changed, and in the other, the size of the donor cells was changed; each protocol was based on our original methodology. Ultimately, we obtained an unexpectedly high rate of adult clone formation using the protocol that differed with respect to the timing of donor nuclei transfer. Specifically, 17% of the transplants that developed to the blastula stage ultimately developed into adult clones. The success rate with this method was 13 times higher than that obtained using the original method. Analyses focusing on the reasons for this high success rate of clone formation will help to elucidate the mechanism of clone formation that occurs with this method.

  12. Analysis of the electrosensory pyramidal cell bursting model for weakly electric fish: model prediction under low levels of dendritic potassium conductance.

    Science.gov (United States)

    Shirahata, T

    2012-09-01

    Pyramidal cells in the electrosensory lateral line lobe (ELL) of weakly electric fish produce burst discharge. A Hodgkin-Huxley-type model, called ghostburster, consisting of two compartments (soma and dendrite) reproduces ELL pyramidal cell bursting observed in vitro. A previous study analyzed the ghostburster by treating Is and gDr,d as bifurcation parameters (Is: current injected into the somatic compartment and gDr,d: maximal conductance of the delayed rectifying potassium current in the dendritic compartment) and indicated that when both Is and gDr,d are set at particular values, the ghostburster shows a codimension-two bifurcation at which both saddle-node bifurcation of fixed points and saddle-node bifurcation of limit cycles occur simultaneously. In the present study, the ghostburster was investigated to clarify the bursting that occurred at gDr,d values smaller than that at the codimension-two bifurcation. Based on the number of spikes per burst, various burst patterns were observed depending on the (Is, gDr,d) values. Depending on the (Is, gDr,d) values, the burst trajectory in a phase space of the ghostburster showed either a high or a low degree of periodicity. Compared to the previous study, the present findings contribute to a more detailed understanding of ghostburster bursting.

  13. Selection and characterization of novel DNA aptamers specifically recognized by Singapore grouper iridovirus-infected fish cells.

    Science.gov (United States)

    Li, Pengfei; Wei, Shina; Zhou, Lingli; Yang, Min; Yu, Yepin; Wei, Jingguang; Jiang, Guohua; Qin, Qiwei

    2015-11-01

    Singapore grouper iridovirus (SGIV) is a major viral pathogen of grouper aquaculture, and has caused heavy economic losses in China and South-east Asia. In this study, we generated four ssDNA aptamers against SGIV-infected grouper spleen (GS) cells using SELEX (systematic evolution of ligands by exponential enrichment) technology. Four aptamers exhibited high affinity to SGIV-infected GS cells, in particular the Q2 aptamer. Q2 had a binding affinity of 12.09 nM, the highest of the four aptamers. These aptamers also recognized SGIV-infected tissues with high levels of specificity. Protease treatment and flow cytometry analysis of SGIV-infected cells revealed that the target molecules of the Q3, Q4 and Q5 aptamers were trypsin-sensitive proteins, whilst the target molecules of Q2 might be membrane lipids or surface proteins that were not trypsin-sensitive. The generated aptamers appeared to inhibit SGIV infection in vitro. Aptamer Q2 conferred the highest levels of protection against SGIV and was able to inhibit SGIV infection in a dose-dependent manner. In addition, Q2 was efficiently internalized by SGIV-infected GS cells and localized at the viral assembly sites. Our results demonstrated that the four novel aptamers we generated were specific for SGIV-infected cells and could potentially be applied as rapid molecular diagnostic test reagents or therapeutic drugs targeting SGIV.

  14. Ependymal cells of the filum terminale in fish (Poecilia sphenops) adapted to freshwater and saltwater: electron microscopic study.

    Science.gov (United States)

    Kriebel, R M

    1981-09-01

    The ependymal lining of the central canal of the filum terminale and spinal cord in the vicinity of the caudal neurosecretory system in P. sphenops was examined in this study. Two general cell types based on shape and location were observed in the ependymal lining: cuboidal ependyma located in dorsal aspects of the filum terminal and columnar to pseudostratified ependymal cells found in ventrolateral and ventral aspects of the filum terminale. Comparison of the ependymal lining was made in animals adapted to saltwater and freshwater. In animals adapted to saltwater there was an increase in the basal infolding of the cell membrane of the dorsal cuboidal ependyma. Infolding of the basal cell membrane is a phenomenon shared by cells known to participate in transport of electrolytes. Since a possible functional relationship between the ependyma of the third ventricle and median eminence has been shown, in future studies on the osmoregulatory function of the caudal neurosecretory system the ependymal lining of the central canal in this region should be considered.

  15. Short communication. Evaluation of presumptive caudal fin blastema cells as candidate donors in intra species zebra fish (Danio rerio) chimaeras

    Energy Technology Data Exchange (ETDEWEB)

    Perez-Camps, M.; Francisco Simao, M.; Cardona Costa, J.; Garcia-Ximenez, F.

    2008-07-01

    The blastema is a regenerative tissue with remarkable pluripotency. The aim of this work done on zebrafish (Danio rerio) was to define technical procedures required for obtaining and integrating blastema cells into embryos at the mid blastula transition stage (MBT) and the effect on survival, as well as the capacity to produce pigmented chimaeras. Wild type blastema cells were injected into gold type MBT embryos (E). Wild MBT blastomere cells were also injected into gold type MBT embryos as a control (C{sub 1}). A second control group, C{sub 2}, was not subjected to any manipulation. Survival was evaluated at 24, 48 and 72 h after performing the chimaerism, and the rate of adult chimaeras evaluated. The results showed significant differences in embryo survival between the E and C{sub 1} groups in embryo survival at 24 and 48 h postchimaerism (24 h: E-83.49% vs C{sub 1}-54.8%, p < 0.05; 48 h: E-98.83% vs C{sub 1}-85.13%, p < 0.05). There was no significant difference, at any time, between E and C{sub 2}. The results at 72 h for E and the controls (E-89.41%; C{sub 1}-84.12% and C{sub 2}-92.55%) indicate that insertion of blastema cells does not have a negative effect on embryo development. The results in adults (E: 0 chimaeras from 7 specimens; C{sub 1}: 5 chimaeras from 17 specimens) suggest that the dedifferentiation grade of the blastema cells may not be enough to generate germ-line chimaeras, but their condition of potentially dedifferentiating cells may be an advantage when using them as donor nuclei in somatic cloning by nuclear transplant. Additional key words: biodiversity, chimaerism, embryo, zebrafish. (Author) 19 refs.

  16. Plastic fish

    CERN Multimedia

    Antonella Del Rosso

    2015-01-01

    In terms of weight, the plastic pollution in the world’s oceans is estimated to be around 300,000 tonnes. This plastic comes from both land-based and ocean-based sources. A lecture at CERN by chemist Wolfgang Trettnak addressed this issue and highlighted the role of art in raising people’s awareness.   Artwork by Wolfgang Trettnak. Packaging materials, consumer goods (shoes, kids’ toys, etc.), leftovers from fishing and aquaculture activities… our oceans and beaches are full of plastic litter. Most of the debris from beaches is plastic bottles. “PET bottles have high durability and stability,” explains Wolfgang Trettnak, a chemist by education and artist from Austria, who gave a lecture on this topic organised by the Staff Association at CERN on 26 May. “PET degrades very slowly and the estimated lifetime of a bottle is 450 years.” In addition to the beach litter accumulated from human use, rivers bring several ki...

  17. Genotoxic effects of selected biocides on RTG-2 fish cells by means of a modified Fast Micromethod Assay.

    Science.gov (United States)

    Sánchez-Fortún, S; Llorente, M T; Castaño, A

    2005-06-01

    A sensitive in vitro assay for detecting DNA damage in RTG-2 cells culture is described. This assay employs a dye, PicoGreen double stranded DNA (dsDNA) quantitation reagent, which becomes intensely fluorescent upon binding nucleic acids. The assay includes a simple and rapid 50-min sample lysis in the presence of EDTA, SDS, and high urea concentration at pH 10, followed by time-dependent DNA denaturation at pH 11.6 after NaOH addition. The time course and the extent of DNA denaturation are followed in a microplate fluorescence reader at room temperature for less than 1h. Comparative studies between suspension and fixed RTG-2 cells indicated that it is possible to apply this methodology in both cases with good results. Neutral red assay was used for to determine the cellular viability when RTG-2 cultures were exposed to tetrakis(hydroxymethyl) phosphonium chloride (THPC) and benzalkonium chloride (BC), as biocides used in the disinfection of cooling towers. The results obtained by neutral red assay indicate IC(50(48)) values of 0.017 (0.011-0.028) and 2.71 (1.91-3.86) mg/L for tetrakis(hydroxymethyl) phosphonium chloride and benzalkonium chloride, respectively. DNA damage has been evaluated for both disinfectants in RTG-2 culture, by exposure to 1/10-, 1/25-, 1/50-, and 1/100-IC(50(48)) value, and the results obtained indicate a strain scission factor (SSF) of 0.126+/-0.014, 0.181+/-0.014, 0.217+/-0.013, and 0.245+/-0.013 in cell suspensions, and 0.077+/-0.019, 0.107+/-0.014, 0.151+/-0.014, and 0.202+/-0.015 in attached cells for tetrakis(hydroxymethyl) phosphonium chloride; while the SSF values for benzalkonium chloride are 0.023+/-0.009, 0.033+/-0.017, 0.068+/-0.012, and 0.088+/-0.015 in cell suspensions, and 0.033+/-0.010, 0.044+/-0.011, 0.080+/-0.009, and 0.093+/-0.010 in attached cells. Thus, the assay proposed in this study has made it possible to show DNA damage in RTG-2 cells when exposed to 0.2(1/100 IC(50(48))) and 300(1/10 IC(50(48))) Hg/L of tetrakis

  18. DEVELOPMENT OF LAW RELATING TO FACTORIES IN INDIA

    Directory of Open Access Journals (Sweden)

    M. K. Sahu

    2015-01-01

    Full Text Available The rapid growth of industrial town and factories has paved the way to develop our industrial legislation accordingly. The Government of India never expressed their interest in framing separate legislation vis-à-vis factories which resulted in implementation of the same statute which was enacted pre-independence. It was done by virtue of Art. 372 of the Constitution of India. However, the Constitutional Lawmakers created vacuum for the implementation of new statute in accordance with the demand of society by inserting scope under the Directive Principles of State Policies. However, in the 67 years history of Indian Republic, there are unprecedented developments of law relating to factories in India.The Government of India, with the adoption of existed statute, made an effort to incorporate the welfare legislation but it never developed along with the change in time. It is to be noticed that as far as existing statutes are concerned, the development is an effect of judicial pronouncement or some tragic incident like Bhopal Gas Tragedy. This paper succinctly describes the history of factory legislation, the constitutional validity of the previous statute and necessary amendment which have already been done and / or on the verge of being amended. It will further discuss contribution of judiciary in developing the law relating to factories, scope of industrial jurisprudence in promoting the development of factory legislation. The primary focus of the research project is to reflect upon the areas where factory legislation has developed, so that proper yardstick could be made in order to put emphasis on those areas which have been remained untouched.

  19. Hydrolyzed fish proteins modulates both inflammatory and antioxidant gene expression as well as protein expression in a co culture model of liver and head kidney cells isolated from Atlantic salmon (Salmo salar).

    Science.gov (United States)

    Holen, Elisabeth; He, Juyun; Araujo, Pedro; Seliussen, Jørgen; Espe, Marit

    2016-07-01

    Hydrolyzed fish proteins (H-pro) contain high concentrations of free amino acids and low molecular peptides that potentially may benefit fish health. The following study aimed to test whether the water-soluble phase of H-pro could attenuate lipopolysaccharide (LPS) provoked inflammation in liver cells and head kidney cells isolated from Atlantic salmon. Cells were grown as mono cultures or co cultures to assess possible crosstalk between immune cells and metabolic cells during treatments. Cells were added media with or without H-pro for 2 days before LPS exposure and harvested 24 h post LPS exposure. Respective cells without H-pro and LPS were used as controls. H-pro alone could affect expression of proteins directly as H-pro increased catalase protein expression in head kidney- and liver cells, regardless of culturing methods and LPS treatment. Leukotriene B4 (LTB4) production was also increased by H-pro in head kidney cells co cultured with liver cells. H-pro increased LPS induced interleukin 1β (IL-1β) transcription in liver cells co cultured with head kidney cells. All cultures of head kidney cells showed a significant increase in IL-1β transcription when treated with H-pro + LPS. H-pro decreased caspase-3 transcription in liver cells cultured co cultured with head kidney cells. Peroxisome proliferator activated receptor α (PPAR α) was upregulated, regardless of treatment, in liver cells co cultured with head kidney cells clearly showing that culturing method alone affected gene transcription. H-pro alone and together with LPS as an inflammation inducer, affect both antioxidant and inflammatory responses.

  20. Temperature dependency of cupular mechanics and hair cell frequency selectivity in the fish canal lateral line organ

    NARCIS (Netherlands)

    Wiersinga-Post, JEC; van Netten, SM

    2000-01-01

    The mechanical frequency selectivity of the cupula located in the supraorbital lateral line canal and the frequency selectivity of the hair cells driven by the cupula were measured simultaneously in vivo. Laser interferometry was used to measure cupular mechanics and extracellular receptor potential

  1. Effects of nutrients and fish on periphyton and plant biomass across a European latitudinal gradient

    NARCIS (Netherlands)

    Bécares, E.; Gomá, J.; Fernández-Aláez, M.; Fernández-Aláez, C.; Romo, S.; Rosa Miracle, M.; Ståhl-Delbanco, A.; Hansson, L-A.; Gyllström, M.; van de Bund, W.; Van Donk, E.; Kairesalo, T.; Hietala, J.; Stephen, D.; Balayla, D.; Moss, B.

    2008-01-01

    Replicated, factorial mesocosm experiments were conducted across Europe to study the effects of nutrient enrichment and fish density on macrophytes and on periphyton chlorophyll a (chl-a) with regard to latitude. Periphyton chl-a densities and plant decline were significantly related to nutrient loa

  2. Incense and Joss Stick Making in Small Household Factories, Thailand

    Directory of Open Access Journals (Sweden)

    S Siripanich

    2014-07-01

    Full Text Available Background: Incense and joss stick are generally used in the world. Most products were made in small household factories. There are many environmental and occupational hazards in these factories.Objective: To evaluate the workplace environmental and occupational hazards in small household incense and joss stick factories in Roi-Et, Thailand.Methods: Nine small household factories in rural areas of Roi-Et, Thailand, were studied. Dust concentration and small aerosol particles were counted through real time exposure monitoring. The inductively coupled plasma optical emission spectrometry (ICP-OES was used for quantitative measurement of heavy metal residue in incense products.Results: Several heavy metals were found in dissolved dye and joss sticks. Those included barium, manganese, and lead. Rolling and shaking processes produced the highest concentration of dust and aerosols. Only 3.9 % of female workers used personal protection equipment.Conclusion: Dust and chemicals were major threats in small household incense and joss stick factories in Thailand. Increasing awareness towards using personal protection equipment and emphasis on elimination of environmental workplace hazards should be considered to help the workers of this industry.

  3. Factorial Schur functions and the Yang-Baxter equation

    CERN Document Server

    Bump, Daniel; Nakasuji, Maki

    2011-01-01

    Factorial Schur functions are generalizations of Schur functions that have, in addition to the usual variables, a second family of "shift" parameters. We show that a factorial Schur function times a deformation of the Weyl denominator may be expressed as the partition function of a particular statistical-mechanical system (six vertex model). The proof is based on the Yang-Baxter equation. There is a deformation parameter $t$ which may be specialized in different ways. If $t=-1$, then we recover the expression of the factorial Schur function as a ratio of alternating polynomials. If $t=0$, we recover the description as a sum over tableaux. If $t=\\infty$ we recover a description of Lascoux that was previously considered by McNamara. We also are able to prove using the Yang-Baxter equation the asymptotic symmetry of the factorial Schur functions in the shift parameters. Finally, we give a proof using our methods of the dual Cauchy identity for factorial Schur functions. Thus using our methods we are able to give...

  4. In vitro cytotoxicity assessment of the biocidal agents sodium o-phenylphenol, sodium o-benzyl-p-chlorophenol, and sodium p-tertiary amylphenol using established fish cell lines.

    Science.gov (United States)

    Davoren, Maria; Fogarty, Andrew M

    2006-10-01

    The cytotoxicity of three biocidal agents frequently employed as active ingredients in phenolic-based disinfectants, were evaluated in three established fish cell lines (EPC, CHSE and RTG-2). Cell viability was assessed using two fluorescent indicator dyes, Alamar Blue for metabolism and neutral red for lysosomal activity. Total protein content was also quantified as a measure of cell detachment. In order to evaluate the sensitivity of the cell cultures, the results obtained were compared with toxicity data obtained from a previous study with the same three compounds and the in vivo lethality test with rainbow trout. Results from this study established that each of the three cell lines ranked the tested chemicals in the same order of toxicity as the in vivo test; however, the cell cultures were found to be an order of magnitude less sensitive than whole fish studies with the same compounds. The chemical sodium o-benzyl-p-chlorophenol was consistently ranked the most toxic of the tested compounds with each cell line and the endpoints employed. The rank order of toxicity was always sodium o-benzyl-p-chlorophenol > sodium p-tertiary amylphenol > sodium o-phenylphenol. The EPC cells were found to be the most sensitive cell line tested based on Alamar Blue IC(50) data, and the Alamar Blue assay was consistently found to be the most sensitive endpoint of the three cytotoxicity assays employed.

  5. Effect of a low-fat fish oil diet on proinflammatory eicosanoids and cell-cycle progression score in men undergoing radical prostatectomy.

    Science.gov (United States)

    Galet, Colette; Gollapudi, Kiran; Stepanian, Sevan; Byrd, Joshua B; Henning, Susanne M; Grogan, Tristan; Elashoff, David; Heber, David; Said, Jonathan; Cohen, Pinchas; Aronson, William J

    2014-01-01

    We previously reported that a 4- to 6-week low-fat fish oil (LFFO) diet did not affect serum insulin-like growth factor (IGF)-1 levels (primary outcome) but resulted in lower omega-6 to omega-3 fatty acid ratios in prostate tissue and lower prostate cancer proliferation (Ki67) as compared with a Western diet. In this post hoc analysis, the effect of the LFFO intervention on serum pro-inflammatory eicosanoids, leukotriene B4 (LTB4) and 15-S-hydroxyeicosatetraenoic acid [15(S)-HETE], and the cell-cycle progression (CCP) score were investigated. Serum fatty acids and eicosanoids were measured by gas chromatography and ELISA. CCP score was determined by quantitative real-time reverse transcriptase PCR (RT-PCR). Associations between serum eicosanoids, Ki67, and CCP score were evaluated using partial correlation analyses. BLT1 (LTB4 receptor) expression was determined in prostate cancer cell lines and prostatectomy specimens. Serum omega-6 fatty acids and 15(S)-HETE levels were significantly reduced, and serum omega-3 levels were increased in the LFFO group relative to the Western diet group, whereas there was no change in LTB4 levels. The CCP score was significantly lower in the LFFO compared with the Western diet group. The 15(S)-HETE change correlated with tissue Ki67 (R = 0.48; P < 0.01) but not with CCP score. The LTB4 change correlated with the CCP score (r = 0.4; P = 0.02) but not with Ki67. The LTB4 receptor BLT1 was detected in prostate cancer cell lines and human prostate cancer specimens. In conclusion, an LFFO diet resulted in decreased 15(S)-HETE levels and lower CCP score relative to a Western diet. Further studies are warranted to determine whether the LFFO diet antiproliferative effects are mediated through the LTB4/BLT1 and 15(S)-HETE pathways.

  6. Three Kinds of Fish

    DEFF Research Database (Denmark)

    Høst, Jeppe Engset

    2012-01-01

    There are three kinds of fish. Fish you were given, fish you bought and fish you lease. This might sound a bit odd, but it is nevertheless the basis for the activities of Danish commercial fishers since the introduction of transferable fishing concessions (TFCs) in 2007. In the current 2012 reform...... of market based systems are wild speculation, concentration and monopolization of fishing access and subsequent leasing with fishing communities and new entrants very likely being worse off (see for example the chapter “From fishing rights to financial derivatives” is this volume or Olson 2011; Sumaila 2010...... will examine five Danish fishing operations and discuss how they have reacted in different ways to the newly introduced system of transferable fishing concessions. By introducing TFCs as a solution to fleet overcapacity, the EU Commission will also be introducing a system where buying, selling and leasing...

  7. CRISPR/Cas9 advances engineering of microbial cell factories

    DEFF Research Database (Denmark)

    Jakociunas, Tadas; Jensen, Michael Krogh; Keasling, Jay D.

    2016-01-01

    interspaced palindromic repeats (CRISPR) and its associated proteins (Cas) have become the method of choice for precision genome engineering in many organisms due to their orthogonality, versatility and efficacy. Here we review the strategies adopted for implementation of RNA-guided CRISPR/Cas9 genome editing......-RNAs will be highlighted. Finally, this review will provide a perspective on the immediate challenges and opportunities foreseen by the use of CRISPR/Cas9 genome engineering and regulation in the context of metabolic engineering....

  8. The methylotrophic yeast Hansenula polymorpha : A versatile cell factory

    NARCIS (Netherlands)

    van Dijk, R.; Faber, Klaas Nico; Kiel, J.A.K.W.; Veenhuis, M.; van der Klei, I.

    2000-01-01

    The development of heterologous overexpression systems for soluble proteins has greatly advanced the study of the structure/function relationships of these proteins and their biotechnological and pharmaceutical applications. In this paper we present an overview on several aspects of the use of the m

  9. Transforming Lactococcus lactis into a microbial cell factory

    DEFF Research Database (Denmark)

    Petersen, Kia Vest

    . To simplify further analysis arcA encoding the arginine deiminase was deleted, thus eliminating the arginine catabolism. We found that in L. lactis KF147 xylose is metabolized through two pathways namely the phosphoketolase pathway and the non-oxidative part of the pentose phosphate pathway. The only products...... the potential of Lactococcus lactis as a platform organism for production of biofuels and-chemicals with a focus on characterization and optimization of the xylose metabolism. The plant isolate L. lactis KF147 was selected as the potential platform organism due to its natural ability to utilize both the pentose...... for the next round of integration. The xylose metabolism in L. lactis KF147 was characterized in a defined medium supplemented with 0.2%, 1% or 3% (w/v) xylose. The defined medium contains free arginine, and it was found that L. lactis KF147 co-metabolizes the arginine through the arginine deiminase pathway...

  10. Redirector: designing cell factories by reconstructing the metabolic objective.

    Directory of Open Access Journals (Sweden)

    Graham Rockwell

    Full Text Available Advances in computational metabolic optimization are required to realize the full potential of new in vivo metabolic engineering technologies by bridging the gap between computational design and strain development. We present Redirector, a new Flux Balance Analysis-based framework for identifying engineering targets to optimize metabolite production in complex pathways. Previous optimization frameworks have modeled metabolic alterations as directly controlling fluxes by setting particular flux bounds. Redirector develops a more biologically relevant approach, modeling metabolic alterations as changes in the balance of metabolic objectives in the system. This framework iteratively selects enzyme targets, adds the associated reaction fluxes to the metabolic objective, thereby incentivizing flux towards the production of a metabolite of interest. These adjustments to the objective act in competition with cellular growth and represent up-regulation and down-regulation of enzyme mediated reactions. Using the iAF1260 E. coli metabolic network model for optimization of fatty acid production as a test case, Redirector generates designs with as many as 39 simultaneous and 111 unique engineering targets. These designs discover proven in vivo targets, novel supporting pathways and relevant interdependencies, many of which cannot be predicted by other methods. Redirector is available as open and free software, scalable to computational resources, and powerful enough to find all known enzyme targets for fatty acid production.

  11. CRISPR/Cas9 advances engineering of microbial cell factories.

    Science.gov (United States)

    Jakočiūnas, Tadas; Jensen, Michael K; Keasling, Jay D

    2016-03-01

    One of the key drivers for successful metabolic engineering in microbes is the efficacy by which genomes can be edited. As such there are many methods to choose from when aiming to modify genomes, especially those of model organisms like yeast and bacteria. In recent years, clustered regularly interspaced palindromic repeats (CRISPR) and its associated proteins (Cas) have become the method of choice for precision genome engineering in many organisms due to their orthogonality, versatility and efficacy. Here we review the strategies adopted for implementation of RNA-guided CRISPR/Cas9 genome editing with special emphasis on their application for metabolic engineering of yeast and bacteria. Also, examples of how nuclease-deficient Cas9 has been applied for RNA-guided transcriptional regulation of target genes will be reviewed, as well as tools available for computer-aided design of guide-RNAs will be highlighted. Finally, this review will provide a perspective on the immediate challenges and opportunities foreseen by the use of CRISPR/Cas9 genome engineering and regulation in the context of metabolic engineering.

  12. Engineering Escherichia coli Cell Factories for n-Butanol Production.

    Science.gov (United States)

    Dong, Hongjun; Zhao, Chunhua; Zhang, Tianrui; Lin, Zhao; Li, Yin; Zhang, Yanping

    2016-01-01

    The production of n-butanol, as a widely applied solvent and potential fuel, is attracting much attention. The fermentative production of butanol coupled with the production of acetone and ethanol by Clostridium (ABE fermentation) was once one of the oldest biotechnological processes, ranking second in scale behind ethanol fermentation. However, there remain problems with butanol production by Clostridium, especially the difficulty in genetically manipulating clostridial strains. In recent years, many efforts have been made to produce butanol using non-native strains. Until now, the most advanced effort was the engineering of the user-friendly and widely studied Escherichia coli for butanol production. This paper reviews the current progress and problems relating to butanol production by engineered E. coli in terms of prediction using mathematical models, pathway construction, novel enzyme replacement, butanol toxicity, and tolerance engineering strategies.

  13. Lactic acid bacterial cell factories for gamma-aminobutyric acid.

    Science.gov (United States)

    Li, Haixing; Cao, Yusheng

    2010-11-01

    Gamma-aminobutyric acid is a non-protein amino acid that is widely present in organisms. Several important physiological functions of gamma-aminobutyric acid have been characterized, such as neurotransmission, induction of hypotension, diuretic effects, and tranquilizer effects. Many microorganisms can produce gamma-aminobutyric acid including bacteria, fungi and yeasts. Among them, gamma-aminobutyric acid-producing lactic acid bacteria have been a focus of research in recent years, because lactic acid bacteria possess special physiological activities and are generally regarded as safe. They have been extensively used in food industry. The production of lactic acid bacterial gamma-aminobutyric acid is safe and eco-friendly, and this provides the possibility of production of new naturally fermented health-oriented products enriched in gamma-aminobutyric acid. The gamma-aminobutyric acid-producing species of lactic acid bacteria and their isolation sources, the methods for screening of the strains and increasing their production, the enzymatic properties of glutamate decarboxylases and the relative fundamental research are reviewed in this article. And the potential applications of gamma-aminobutyric acid-producing lactic acid bacteria were also referred to.

  14. Engineering a cyanobacterial cell factory for production of lactic acid.

    NARCIS (Netherlands)

    Angermayr, S.A.; Paszota, M.; Hellingwerf, K.J.

    2012-01-01

    Metabolic engineering of microorganisms has become a versatile tool to facilitate production of bulk chemicals, fuels, etc. Accordingly, CO(2) has been exploited via cyanobacterial metabolism as a sustainable carbon source of biofuel and bioplastic precursors. Here we extended these observations by

  15. Post-production protein stability: trouble beyond the cell factory

    Directory of Open Access Journals (Sweden)

    Vazquez Esther

    2011-08-01

    Full Text Available Abstract Being protein function a conformation-dependent issue, avoiding aggregation during production is a major challenge in biotechnological processes, what is often successfully addressed by convenient upstream, midstream or downstream approaches. Even when obtained in soluble forms, proteins tend to aggregate, especially if stored and manipulated at high concentrations, as is the case of protein drugs for human therapy. Post-production protein aggregation is then a major concern in the pharmaceutical industry, as protein stability, pharmacokinetics, bioavailability, immunogenicity and side effects are largely dependent on the extent of aggregates formation. Apart from acting at the formulation level, the recombinant nature of protein drugs allows intervening at upstream stages through protein engineering, to produce analogue protein versions with higher stability and enhanced therapeutic values.

  16. Lactic acid bacteria as a cell factory for riboflavin production.

    Science.gov (United States)

    Thakur, Kiran; Tomar, Sudhir Kumar; De, Sachinandan

    2016-07-01

    Consumers are increasingly becoming aware of their health and nutritional requirements, and in this context, vitamins produced in situ by microbes may suit their needs and expectations. B groups vitamins are essential components of cellular metabolism and among them riboflavin is one of the vital vitamins required by bacteria, plants, animals and humans. Here, we focus on the importance of microbial production of riboflavin over chemical synthesis. In addition, genetic abilities for riboflavin biosynthesis by lactic acid bacteria are discussed. Genetically modified strains by employing genetic engineering and chemical analogues have been developed to enhance riboflavin production. The present review attempts to collect the currently available information on riboflavin production by microbes in general, while placing greater emphasis on food grade lactic acid bacteria and human gut commensals. For designing riboflavin-enriched functional foods, proper selection and exploitation of riboflavin-producing lactic acid bacteria is essential. Moreover, eliminating the in situ vitamin fortification step will decrease the cost of food production.

  17. MudPIT analysis of alkaline tolerance by Listeria monocytogenes strains recovered as persistent food factory contaminants.

    Science.gov (United States)

    Nilsson, Rolf E; Latham, Roger; Mellefont, Lyndal; Ross, Tom; Bowman, John P

    2012-05-01

    Alkaline solutions are used to clean food production environments but the role of alkaline resistance in persistent food factory contamination by Listeria monocytogenes is unknown. We used shotgun proteomics to characterise alkaline adapted L. monocytogenes recovered as persistent and transient food factory contaminants. Three unrelated strains were studied including two persistent and a transient food factory contaminant determined using multilocus sequence typing (MLST). The strains were adapted to growth at pH 8.5 and harvested in exponential phase. Protein extracts were analysed using multidimensional protein identification technology (MudPIT) and protein abundance compared by spectra counting. The strains elicited core responses to alkaline growth including modulation of intracellular pH, stabilisation of cellular processes and reduced cell-division, independent to lineage, MLST or whether the strains were transient or persistent contaminants. Alkaline adaptation by all strains corresponded to that expected in stringent-response induced cells, with protein expression supporting metabolic shifts concordant with elevated alarmone production and indicating that the alkaline-stringent response results from energy rather than nutrient limitation. We believe this is the first report describing induction of a stringent response in different L. monocytogenes strains by alkaline pH under non-limiting growth conditions. The work emphasises the need for early intervention to avoid persistent food factory contamination by L. monocytogenes.

  18. Comparative genotoxicity testing of Rhine river sediment extracts using the comet assay with permanent fish cell lines (RTG-2 and RTL-W1) and the Ames test

    Energy Technology Data Exchange (ETDEWEB)

    Kosmehl, T.; Braunbeck, T.; Hollert, H. [Dept. of Zoology, Aquatic Ecology and Toxicology Section, Univ. of Heidelberg (Germany); Krebs, F.; Manz, W. [German Federal Inst. of Hydrology, Koblenz (Germany); Erdinger, L. [Dept. for Hygiene and Medical Microbiology, Inst. for Hygiene, Univ. of Heidelberg (Germany)

    2004-07-01

    Whilst at least in Germany assessment strategies on the basis of chemical analysis and acute toxicity data dominated the last decades, the development of more specific biological endpoints and biomarkers in ecotoxicology is required in order to arrive at a good ecological potential and good chemical status of surface waters in the European river basins until the year 2015, as required by the European Water Framework Directive. Since sediments have for long been known to function both as a sink and as a source of pollutants in aquatic systems, and since part of the particle-associated substances have frequently been demonstrated to cause mutagenic and carcinogenic effects in aquatic organisms, particularly in fish, there is, among other requirements, an urgent need to develop, standardize and implement integrated vertebrate-based test systems addressing genotoxicity into recent sediment investigation strategies. Thus, the present study was designed to compare the suitability of two commonly used test systems, the comet assay and the Ames test, for the evaluation of the ecotoxicological burden of surface and core sediment samples from the river Rhine. Methods (or main features). In order to determine the importance of inherent enzymatic activities, two permanent fish cell lines with different biotransformation capacities, RTL-W1 and RTG-2, were compared with respect to their capability of detecting genotoxic effects in 18 surface and core sediment samples from 9 locations along the river Rhine in the comet assay with and without exogenous bioactivation. For further comparison, as a prokaryotic mutagenicity assay, the Salmonella plate incorporation assay (Ames test) with the test strains TA98 and TA100 with and without exogenous metabolic activation was used. Results and discussion. Whereas all sediment extracts induced genotoxic effects in the comet assay with RTL-W1 cells, only 12 out of 18 sediment extracts revealed significant genotoxicity in the tests with the

  19. Longitudinal Beam Stability in the SUPER B-FACTORY

    Energy Technology Data Exchange (ETDEWEB)

    Novokhatski, A.; /SLAC; Zobov, M.; /Frascati

    2009-07-06

    We give an overview of wake fields and impedances in a proposed Super B project, which is based on extremely low emittance beams colliding at a large angle with a crab waist transformation. Understanding the effects that wake fields have on the beam is critical for a successful machine operation. We use our combined experience from the operation of the SLAC B-factory and DA{Phi}NE {Phi}-factory to eliminate strong HOM sources and minimize the chamber impedance in the Super B design. Based on a detailed study of the wake fields in this design we have developed a quasi-Green's function for the entire ring that is used to study bunch lengthening and beam stability. In particular, we check the stability threshold using numerical solutions of the Fokker-Plank equation. We also make a comparison of numerical simulations with the bunch lengthening data in the B- factory.

  20. Summation of Divergent Power Series by Means of Factorial Series

    CERN Document Server

    Weniger, Ernst Joachim

    2010-01-01

    Factorial series played a major role in Stirling's classic book "Methodus Differentialis" (1730), but now only a few specialists still use them. This article wants to show that this neglect is unjustified, and that factorial series are useful numerical tools for the summation of divergent (inverse) power series. This is documented by summing the divergent asymptotic expansion for the exponential integral $E_{1} (z)$ and the factorially divergent Rayleigh-Schr\\"{o}dinger perturbation expansion for the quartic anharmonic oscillator. Stirling numbers play a key role since they occur as coefficients in expansions of an inverse power in terms of inverse Pochhammer symbols and vice versa. It is shown that the relationships involving Stirling numbers are special cases of more general orthogonal and triangular transformations.