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Sample records for hybridization fish experiments

  1. Marine Fish Hybridization

    KAUST Repository

    He, Song

    2017-04-01

    Natural hybridization is reproduction (without artificial influence) between two or more species/populations which are distinguishable from each other by heritable characters. Natural hybridizations among marine fishes were highly underappreciated due to limited research effort; it seems that this phenomenon occurs more often than is commonly recognized. As hybridization plays an important role in biodiversity processes in the marine environment, detecting hybridization events and investigating hybridization is important to understand and protect biodiversity. The first chapter sets the framework for this disseration study. The Cohesion Species Concept was selected as the working definition of a species for this study as it can handle marine fish hybridization events. The concept does not require restrictive species boundaries. A general history and background of natural hybridization in marine fishes is reviewed during in chapter as well. Four marine fish hybridization cases were examed and documented in Chapters 2 to 5. In each case study, at least one diagnostic nuclear marker, screened from among ~14 candidate markers, was found to discriminate the putative hybridizing parent species. To further investigate genetic evidence to support the hybrid status for each hybrid offspring in each case, haploweb analysis on diagnostic markers (nuclear and/or mitochondrial) and the DAPC/PCA analysis on microsatellite data were used. By combining the genetic evidences, morphological traits, and ecological observations together, the potential reasons that triggered each hybridization events and the potential genetic/ecology effects could be discussed. In the last chapter, sequences from 82 pairs of hybridizing parents species (for which COI barcoding sequences were available either on GenBank or in our lab) were collected. By comparing the COI fragment p-distance between each hybridizing parent species, some general questions about marine fish hybridization were discussed: Is

  2. Standardization and optimization of fluorescence in situ hybridization (FISH for HER-2 assessment in breast cancer: A single center experience

    Directory of Open Access Journals (Sweden)

    Magdalena Bogdanovska-Todorovska

    2018-05-01

    Full Text Available Accurate assessment of human epidermal growth factor receptor 2 (HER-2 is crucial in selecting patients for targeted therapy. Commonly used methods for HER-2 testing are immunohistochemistry (IHC and fluorescence in situ hybridization (FISH. Here we presented the implementation, optimization and standardization of two FISH protocols using breast cancer samples and assessed the impact of pre-analytical and analytical factors on HER-2 testing. Formalin fixed paraffin embedded (FFPE tissue samples from 70 breast cancer patients were tested for HER-2 using PathVysion™ HER-2 DNA Probe Kit and two different paraffin pretreatment kits, Vysis/Abbott Paraffin Pretreatment Reagent Kit (40 samples and DAKO Histology FISH Accessory Kit (30 samples. The concordance between FISH and IHC results was determined. Pre-analytical and analytical factors (i.e., fixation, baking, digestion, and post-hybridization washing affected the efficiency and quality of hybridization. The overall hybridization success in our study was 98.6% (69/70; the failure rate was 1.4%. The DAKO pretreatment kit was more time-efficient and resulted in more uniform signals that were easier to interpret, compared to the Vysis/Abbott kit. The overall concordance between IHC and FISH was 84.06%, kappa coefficient 0.5976 (p < 0.0001. The greatest discordance (82% between IHC and FISH was observed in IHC 2+ group. A standardized FISH protocol for HER-2 assessment, with high hybridization efficiency, is necessary due to variability in tissue processing and individual tissue characteristics. Differences in the pre-analytical and analytical steps can affect the hybridization quality and efficiency. The use of DAKO pretreatment kit is time-saving and cost-effective.

  3. Parasitic fauna in hybrid tambacu from fish farms

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    Ronilson Macedo Silva

    2013-08-01

    Full Text Available The objective of this work was to evaluate the parasitic fauna of hybrid tambacu (Colossoma macropomum x Piaractus mesopotamicus from fish farms and the host-parasite relationship. A hundred and fourteen fish were collected from four fish farms in Macapá, in the state of Amapá, Brazil, 80.7% of which were infected by: Ichthyophthirius multifiliis (Ciliophora; Piscinoodinium pillulare (Dinoflagellida; Anacanthorus spatulatus, Notozothecium janauachensis, and Mymarothecium viatorum (Monogenoidea; Neoechinorhynchus buttnerae (Acanthocephala; Cucullanus colossomi (Nematoda; Perulernaea gamitanae (Lernaeidae; and Proteocephalidae larvae (Cestoda. A total of 8,136,252 parasites were collected from the examined fish. This is the first record of N. buttnerae, C. colossomi, N. janauachensis, M. viatorum, and Proteocephalidae for hybrid tambacu in Brazil. Ichthyophthirius multifiliis was the most prevalent parasite, whereas endohelminths were the less. A positive correlation was observed between number of I. multifiliis and total length and weight of fish, as well as between number of P. gamitanae and total length. The infection by I. multifiliis had association with the parasitism by Monogenoidea. Low water quality contributes to high parasitism of hybrid tambacu by ectoparasites, which, however, does not influence the relative condition factor of fish.

  4. The chromosomal constitution of fish hybrid lineage revealed by 5S rDNA FISH.

    Science.gov (United States)

    Zhang, Chun; Ye, Lihai; Chen, Yiyi; Xiao, Jun; Wu, Yanhong; Tao, Min; Xiao, Yamei; Liu, Shaojun

    2015-12-03

    The establishment of the bisexual fertile fish hybrid lineage including the allodiploid and allotetraploid hybrids, from interspecific hybridization of red crucian carp (Carassius auratus red var. 2n = 100, 2n = AA) (♀) × common carp (Cyprinus carpio L. 2n = 100, 2n = BB) (♂), provided a good platform to investigate genetic relationship between the parents and their hybrid progenies. The chromosomal inheritance of diploid and allotetraploid hybrid progenies in successive generations, was studied by applying 5S rDNA fluorescence in situ hybridization. Signals of 5S rDNA distinguished the chromosomal constitution of common carp (B-genome) from red crucian carp (A-genome), in which two strong signals were observed on the first submetacentric chromosome, while no major signal was found in common carp. After fish hybridization, one strong signal of 5S rDNA was detected in the same locus on the chromosome of diploid hybrids. As expected, two strong signals were observed in 4nF3 tetraploid hybrids offspring and it is worth mentioning that two strong signals were detected in a separating bivalent of a primary spermatocyte in 4nF3. Furthermore, the mitosis of heterozygous chromosomes was shown normal and stable with blastular tissue histological studies. We revealed that 5S rDNA signal can be applied to discern A-genome from B-genome, and that 5S rDNA bearing chromosomes can be stably passed down in successive generations. Our work provided a significant method in fish breeding and this is important for studies in fish evolutionary biology.

  5. Standardization and optimization of fluorescence in situ hybridization (FISH) for HER-2 assessment in breast cancer: A single center experience.

    Science.gov (United States)

    Bogdanovska-Todorovska, Magdalena; Petrushevska, Gordana; Janevska, Vesna; Spasevska, Liljana; Kostadinova-Kunovska, Slavica

    2018-05-20

    Accurate assessment of human epidermal growth factor receptor 2 (HER-2) is crucial in selecting patients for targeted therapy. Commonly used methods for HER-2 testing are immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Here we presented the implementation, optimization and standardization of two FISH protocols using breast cancer samples and assessed the impact of pre-analytical and analytical factors on HER-2 testing. Formalin fixed paraffin embedded (FFPE) tissue samples from 70 breast cancer patients were tested for HER-2 using PathVysion™ HER-2 DNA Probe Kit and two different paraffin pretreatment kits, Vysis/Abbott Paraffin Pretreatment Reagent Kit (40 samples) and DAKO Histology FISH Accessory Kit (30 samples). The concordance between FISH and IHC results was determined. Pre-analytical and analytical factors (i.e., fixation, baking, digestion, and post-hybridization washing) affected the efficiency and quality of hybridization. The overall hybridization success in our study was 98.6% (69/70); the failure rate was 1.4%. The DAKO pretreatment kit was more time-efficient and resulted in more uniform signals that were easier to interpret, compared to the Vysis/Abbott kit. The overall concordance between IHC and FISH was 84.06%, kappa coefficient 0.5976 (p characteristics. Differences in the pre-analytical and analytical steps can affect the hybridization quality and efficiency. The use of DAKO pretreatment kit is time-saving and cost-effective.

  6. 21 CFR 866.4700 - Automated fluorescence in situ hybridization (FISH) enumeration systems.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated fluorescence in situ hybridization (FISH... Laboratory Equipment and Reagents § 866.4700 Automated fluorescence in situ hybridization (FISH) enumeration... Hybridization (FISH) Enumeration Systems.” See § 866.1(e) for the availability of this guidance document. [70 FR...

  7. Application of hybrid artificial fish swarm algorithm based on similar fragments in VRP

    Science.gov (United States)

    Che, Jinnuo; Zhou, Kang; Zhang, Xueyu; Tong, Xin; Hou, Lingyun; Jia, Shiyu; Zhen, Yiting

    2018-03-01

    Focused on the issue that the decrease of convergence speed and the precision of calculation at the end of the process in Artificial Fish Swarm Algorithm(AFSA) and instability of results, a hybrid AFSA based on similar fragments is proposed. Traditional AFSA enjoys a lot of obvious advantages in solving complex optimization problems like Vehicle Routing Problem(VRP). AFSA have a few limitations such as low convergence speed, low precision and instability of results. In this paper, two improvements are introduced. On the one hand, change the definition of the distance for artificial fish, as well as increase vision field of artificial fish, and the problem of speed and precision can be improved when solving VRP. On the other hand, mix artificial bee colony algorithm(ABC) into AFSA - initialize the population of artificial fish by the ABC, and it solves the problem of instability of results in some extend. The experiment results demonstrate that the optimal solution of the hybrid AFSA is easier to approach the optimal solution of the standard database than the other two algorithms. In conclusion, the hybrid algorithm can effectively solve the problem that instability of results and decrease of convergence speed and the precision of calculation at the end of the process.

  8. Fish welfare: Fish capacity to experience pain

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    Vučinić Marijana

    2009-01-01

    Full Text Available Teleost fish possess similar nociceptive processing systems to those found in terrestrial vertebrates. It means that they react to potential painful stimuli in a similar manner as mammals and birds. However, the welfare of fish has been the focus of less research than that of higher vertebrates. Humans may affect the welfare of fish through fisheries, aquaculture and a number of other activities. There is scientific evidence to support the assumption that fish have the capacity to experience pain because they possess functional nociceptors, endogenous opioids and opioid receptors, brain structures involved in pain processing and pathways leading from nociceptors to higher brain structures. Also, it is well documented that some anaesthetics and analgesics may reduce nociceptive responses in fish. Behavioural indicators in fish such as lip-rubbing and rocking behaviours are the best proof that fish react to potential painful stimuli. This paper is an overview of some scientific evidence on fish capacity to experience pain.

  9. A simple protocol for attenuating the auto-fluorescence of cyanobacteria for optimized fluorescence in situ hybridization (FISH) imaging.

    Science.gov (United States)

    Zeller, Perrine; Ploux, Olivier; Méjean, Annick

    2016-03-01

    Cyanobacteria contain pigments, which generate auto-fluorescence that interferes with fluorescence in situ hybridization (FISH) imaging of cyanobacteria. We describe simple chemical treatments using CuSO4 or H2O2 that significantly reduce the auto-fluorescence of Microcystis strains. These protocols were successfully applied in FISH experiments using 16S rRNA specific probes and filamentous cyanobacteria. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Widespread hybridization and bidirectional introgression in sympatric species of coral reef fish

    KAUST Repository

    Harrison, Hugo B.

    2017-10-28

    Coral reefs are highly diverse ecosystems, where numerous closely related species often coexist. How new species arise and are maintained in these high geneflow environments have been long-standing conundrums. Hybridization and patterns of introgression between sympatric species provide a unique insight into the mechanisms of speciation and the maintenance of species boundaries. In this study, we investigate the extent of hybridization between two closely related species of coral reef fish: the common coral trout (Plectropomus leopardus) and the bar-cheek coral trout (Plectropomus maculatus). Using a complementary set of 25 microsatellite loci, we distinguish pure genotype classes from first- and later-generation hybrids, identifying 124 interspecific hybrids from a collection of 2,991 coral trout sampled in inshore and mid-shelf reefs of the southern Great Barrier Reef. Hybrids were ubiquitous among reefs, fertile and spanned multiple generations suggesting both ecological and evolutionary processes are acting to maintain species barriers. We elaborate on these finding to investigate the extent of genomic introgression and admixture from 2,271 SNP loci recovered from a ddRAD library of pure and hybrid individuals. An analysis of genomic clines on recovered loci indicates that 261 SNP loci deviate from a model of neutral introgression, of which 132 indicate a pattern of introgression consistent with selection favouring both hybrid and parental genotypes. Our findings indicate genome-wide, bidirectional introgression between two sympatric species of coral reef fishes and provide further support to a growing body of evidence for the role of hybridization in the evolution of coral reef fishes.

  11. Introgressive hybridization in Iberian cyprinid fishes:a cytogenomic approach to homoploid Leuciscinae

    OpenAIRE

    Pereira, Carla Sofia Alves, 1983-

    2013-01-01

    Tese de doutoramento, Biologia (Biologia Evolutiva), Universidade de Lisboa, Faculdade de Ciências, 2013 Hybridization is currently a well-recognized process amongst animals responsible for biodiversity, evolution and speciation processes while defying most species concepts. Hybridization is prevalent among fishes, particularly cyprinids, which therefore constitute good models of study (1) to access general patterns of genomic variation, (2) to identify the genetic basis and the evolutiona...

  12. Utility of chromogenic in situ hybridization (CISH) for detection of EGFR amplification in glioblastoma: comparison with fluorescence in situ hybridization (FISH).

    Science.gov (United States)

    Fischer, Ingeborg; de la Cruz, Clarissa; Rivera, Andreana L; Aldape, Kenneth

    2008-12-01

    In this study, we test the reliability of chromogenic in situ hybridization (CISH) for the detection of epidermal growth factor receptor (EGFR) gene amplification in glioblastoma. Earlier reports have described EGFR CISH in glioblastoma multiforme, but a comparison of CISH with a "gold standard" testing method, such as fluorescence in situ hybridization (FISH), has not been described. Therapies targeting the EGFR-signaling pathway might increase the importance of assessment of EGFR-amplification status. CISH is a potential alternative to FISH as a testing method. To test its reliability, EGFR-amplification status by CISH was assessed in 89 cases of glioblastoma and compared with FISH results, and correlated with the protein expression using immunohistochemistry (IHC) for EGFR. FISH was scored as being EGFR-amplified in 47/89 tumors, CISH as being amplified in 43/89 tumors. The CISH and FISH results were in agreement in 83/89 cases (93%). Four glioblastomas were scored as being amplified by FISH, but not by CISH; whereas amplification was detected in 2 tumors by CISH that were not amplified using FISH. Forty-eight of the 89 cases were positive for EGFR expression by IHC. EGFR amplification was highly correlated with protein expression by IHC, as 40/48 (83%) EGFR IHC-positive cases were found to be EGFR-amplified. The high concordance of CISH and FISH for the assessment of EGFR gene-amplification status indicates that CISH is a viable alternative to FISH for the detection of EGFR gene amplification in glioblastoma. Detectable EGFR expression by IHC can occur in the absence of gene amplification, but is uncommon.

  13. Filter-Adapted Fluorescent In Situ Hybridization (FA-FISH) for Filtration-Enriched Circulating Tumor Cells.

    Science.gov (United States)

    Oulhen, Marianne; Pailler, Emma; Faugeroux, Vincent; Farace, Françoise

    2017-01-01

    Circulating tumor cells (CTCs) may represent an easily accessible source of tumor material to assess genetic aberrations such as gene-rearrangements or gene-amplifications and screen cancer patients eligible for targeted therapies. As the number of CTCs is a critical parameter to identify such biomarkers, we developed fluorescent in situ hybridization (FISH) for CTCs enriched on filters (filter-adapted-FISH, FA-FISH). Here, we describe the FA-FISH protocol, the combination of immunofluorescent staining (DAPI/CD45) and FA-FISH techniques, as well as the semi-automated microscopy method that we developed to improve the feasibility and reliability of FISH analyses in filtration-enriched CTC.

  14. Detection of airborne Legionella while showering using liquid impingement and fluorescent in situ hybridization (FISH).

    Science.gov (United States)

    Deloge-Abarkan, Magali; Ha, Thi-Lan; Robine, Enric; Zmirou-Navier, Denis; Mathieu, Laurence

    2007-01-01

    Aerosols of water contaminated with Legionella bacteria constitute the only mode of exposure for humans. However, the prevention strategy against this pathogenic bacteria risk is managed through the survey of water contamination. No relationship linked the Legionella bacteria water concentration and their airborne abundance. Therefore, new approaches in the field of the metrological aspects of Legionella bioaerosols are required. This study was aimed at testing the main principles for bioaerosol collection (solid impaction, liquid impingement and filtration) and the in situ hybridization (FISH) method, both in laboratory and field assays, with the intention of applying such methodologies for airborne Legionella bacteria detection while showering. An aerosolization chamber was developed to generate controlled and reproducible L. pneumophila aerosols. This tool allowed the identification of the liquid impingement method as the most appropriate one for collecting airborne Legionella bacteria. The culturable fraction of airborne L. pneumophila recovered with the liquid impingement principle was 4 and 700 times higher compared to the impaction and filtration techniques, respectively. Moreover, the concentrations of airborne L. pneumophila in the impinger fluid were on average 7.0 x 10(5) FISH-cells m(-3) air with the fluorescent in situ hybridization (FISH) method versus 9.0 x 10(4) CFU m(-3) air with the culture method. These results, recorded under well-controlled conditions, were confirmed during the field experiments performed on aerosols generated by hot water showers in health institutions. This new approach may provide a more accurate characterization of aerobiocontamination by Legionella bacteria.

  15. Intragenus (Campylomormyrus) and intergenus hybrids in mormyrid fish: Physiological and histological investigations of the electric organ ontogeny.

    Science.gov (United States)

    Kirschbaum, Frank; Nguyen, Linh; Baumgartner, Stephanie; Chi, Hiu Wan Linda; Wolfart, Rene; Elarbani, Khouloud; Eppenstein, Hari; Korniienko, Yevheniia; Guido-Böhm, Lilian; Mamonekene, Victor; Vater, Marianne; Tiedemann, Ralph

    2016-10-01

    African weakly electric mormyrid fish show a high diversity of their electric organ discharge (EOD) both across and within genera. Thanks to a recently developed technique of artificial reproduction in mormyrid fish, we were able to perform hybridizations between different genera and within one genus (Campylomormyrus). The hybrids of intergenus hybridizations exhibited different degrees of reduced survival related to the phylogenetic distance of the parent species: hybrids of the crosses between C. rhynchophorus and its sister genus Gnathonemus survived and developed normally. Hybrids between C. rhynchophorus and a Mormyrus species (a more basal clade compared to Campylomormyrus s) survived up to 42days and developed many malformations, e.g., at the level of the unpaired fins. Hybrids between C. numenius and Hippopotamyrus pictus (a derived clade, only distantly related to Campylomormyrus) only survived for two days during embryological development. Eight different hybrid combinations among five Campylomormyrus species (C. tamandua, C. compressirostris, C. tshokwe, C. rhynchophorus, C. numenius) were performed. The aim of the hybridizations was to combine species with (1) either caudal or rostral position of the main stalk innervating the electrocytes in the electric organ and (2) short, median or long duration of their EOD. The hybrids, though they are still juveniles, show very interesting features concerning electrocyte geometry as well as EOD form and duration: the caudal position of the stalk is prevailing over the rostral position, and the penetration of the stalk is dominant over the non-penetrating feature (in the Campylomormyrus hybrids); in the hybrid between C. rhynchophorus and Gnathonemus petersii it is the opposite. When crossing species with long and short EODs, it is always the long duration EOD that is expressed in the hybrids. The F1-Hybrids of the cross C. tamandua×C. compressirostris are fertile: viable F2-fish could be obtained with artificial

  16. Metaphase FISH on a Chip: Miniaturized Microfluidic Device for Fluorescence in situ Hybridization

    Directory of Open Access Journals (Sweden)

    Niels Tommerup

    2010-11-01

    Full Text Available Fluorescence in situ Hybridization (FISH is a major cytogenetic technique for clinical genetic diagnosis of both inherited and acquired chromosomal abnormalities. Although FISH techniques have evolved and are often used together with other cytogenetic methods like CGH, PRINS and PNA-FISH, the process continues to be a manual, labour intensive, expensive and time consuming technique, often taking over 3–5 days, even in dedicated labs. We have developed a novel microFISH device to perform metaphase FISH on a chip which overcomes many shortcomings of the current laboratory protocols. This work also introduces a novel splashing device for preparing metaphase spreads on a microscope glass slide, followed by a rapid adhesive tape-based bonding protocol leading to rapid fabrication of the microFISH device. The microFISH device allows for an optimized metaphase FISH protocol on a chip with over a 20-fold reduction in the reagent volume. This is the first demonstration of metaphase FISH on a microfluidic device and offers a possibility of automation and significant cost reduction of many routine diagnostic tests of genetic anomalies.

  17. 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.

  18. Does interspecies hybridization affect the host specificity of parasites in cyprinid fish?

    Czech Academy of Sciences Publication Activity Database

    Šimková, A.; Dávidová, M.; Papoušek, Ivo; Vetešník, Lukáš

    2013-01-01

    Roč. 6, č. 95 (2013), s. 95 ISSN 1756-3305 R&D Projects: GA ČR GAP505/12/0375 Institutional support: RVO:68081766 Keywords : Cyprinid fish * Interspecies hybridization * Metazoan parasites * Monogenea * Host specificity Subject RIV: EG - Zoology Impact factor: 3.251, year: 2013

  19. Design of Hybrid Solar and Wind Energy Harvester for Fishing Boat

    Science.gov (United States)

    Banjarnahor, D. A.; Hanifan, M.; Budi, E. M.

    2017-07-01

    In southern beach of West Java, Indonesia, there are many villagers live as fishermen. They use small boats for fishing, in one to three days. Therefore, they need a fish preservation system. Fortunately, the area has high potential of solar and wind energy. This paper presents the design of a hybrid solar and wind energy harvester to power a refrigerator in the fishing boat. The refrigerator should keep the fish in 2 - 4 °C. The energy needed is 720 Wh daily. In the area, the daily average wind velocity is 4.27 m/s and the sun irradiation is 672 W/m2. The design combined two 100W solar panels and a 300W wind turbine. The testing showed that the solar panels can harvest 815 - 817 Wh of energy, while the wind turbine can harvest 43 - 62 Wh of energy daily. Therefore, the system can fulfil the energy requirement in fishing boat, although the solar panels were more dominant. To install the wind turbine on the fishing-boat, a computational design had been conducted. The boat hydrostatic dimension was measured to determine its stability condition. To reach a stable equilibrium condition, the wind turbine should be installed no more than 1.7 m of height.

  20. mathFISH, a web tool that uses thermodynamics-based mathematical models for in silico evaluation of oligonucleotide probes for fluorescence in situ hybridization.

    Science.gov (United States)

    Yilmaz, L Safak; Parnerkar, Shreyas; Noguera, Daniel R

    2011-02-01

    Mathematical models of RNA-targeted fluorescence in situ hybridization (FISH) for perfectly matched and mismatched probe/target pairs are organized and automated in web-based mathFISH (http://mathfish.cee.wisc.edu). Offering the users up-to-date knowledge of hybridization thermodynamics within a theoretical framework, mathFISH is expected to maximize the probability of success during oligonucleotide probe design.

  1. Application of Fluorescence In Situ Hybridization (FISH) Technique for the Detection of Genetic Aberration in Medical Science

    Science.gov (United States)

    Ratan, Zubair Ahmed; Zaman, Sojib Bin; Haidere, Mohammad Faisal; Runa, Nusrat Jahan; Akter, Nasrin

    2017-01-01

    Fluorescence in situ hybridization (FISH) is a macromolecule recognition technique, which is considered as a new advent in the field of cytology. Initially, it was developed as a physical mapping tool to delineate genes within chromosomes. The accuracy and versatility of FISH were subsequently capitalized upon in biological and medical research. This visually appealing technique provides an intermediate degree of resolution between DNA analysis and chromosomal investigations. FISH consists of a hybridizing DNA probe, which can be labeled directly or indirectly. In the case of direct labeling, fluorescent nucleotides are used, while indirect labeling is incorporated with reporter molecules that are subsequently detected by fluorescent antibodies or other affinity molecules. FISH is applied to detect genetic abnormalities that include different characteristic gene fusions or the presence of an abnormal number of chromosomes in a cell or loss of a chromosomal region or a whole chromosome. It is also applied in different research applications, such as gene mapping or the identification of novel oncogenes. This article reviews the concept of FISH, its application, and its advantages in medical science.  PMID:28690958

  2. Influence of environment and mitochondrial heritage on the ecological characteristics of fish in a hybrid zone.

    Directory of Open Access Journals (Sweden)

    Nicolas Stolzenberg

    2009-06-01

    Full Text Available Ecological characteristics (growth, morphology, reproduction arise from the interaction between environmental factors and genetics. Genetic analysis of individuals' life history traits might be used to improve our understanding of mechanisms that form and maintain a hybrid zone.A fish hybrid zone was used to characterize the process of natural selection. Data were collected during two reproductive periods (2001 and 2002 and 1117 individuals (nase, Chondrostama nasus nasus, sofie C. toxostoma toxostoma and hybrids were sampled. Reproductive dates of the two parental species overlapped at sympatric sites. The nase had an earlier reproductive period than the sofie; males had longer reproductive periods for both species. Hybridisation between female nase and male sofie was the most likely. Hybrids had a reproductive period similar to the inherited parental mitochondrial type. Growth and reproductive information from different environments has been synthesised following a bayesian approach of the von Bertalanffy model. Hybrid life history traits appear to link with maternal heritage. Hybrid size from the age of two and size at first maturity appeared to be closer to the size of the maternal origin species (nase or sofie. Median growth rates for hybrids were similar and intermediate between those of the parental species. We observed variable life history traits for hybrids and pure forms in the different parts of the hybrid zone. Geometrical analysis of the hybrid fish shape gave evidence of two main morphologies with a link to maternal heritage.Selective mating seemed to be the underlying process which, with mitochondrial heritage, could explain the evolution of the studied hybrid zone. More generally, we showed the importance of studies on hybrid zones and specifically the study of individuals' ecological characteristics, to improve our understanding of speciation.

  3. Influence of environment and mitochondrial heritage on the ecological characteristics of fish in a hybrid zone.

    Science.gov (United States)

    Stolzenberg, Nicolas; Nguyen The, Bénédicte; Salducci, Marie Dominique; Cavalli, Laurent

    2009-06-18

    Ecological characteristics (growth, morphology, reproduction) arise from the interaction between environmental factors and genetics. Genetic analysis of individuals' life history traits might be used to improve our understanding of mechanisms that form and maintain a hybrid zone. A fish hybrid zone was used to characterize the process of natural selection. Data were collected during two reproductive periods (2001 and 2002) and 1117 individuals (nase, Chondrostama nasus nasus, sofie C. toxostoma toxostoma and hybrids) were sampled. Reproductive dates of the two parental species overlapped at sympatric sites. The nase had an earlier reproductive period than the sofie; males had longer reproductive periods for both species. Hybridisation between female nase and male sofie was the most likely. Hybrids had a reproductive period similar to the inherited parental mitochondrial type. Growth and reproductive information from different environments has been synthesised following a bayesian approach of the von Bertalanffy model. Hybrid life history traits appear to link with maternal heritage. Hybrid size from the age of two and size at first maturity appeared to be closer to the size of the maternal origin species (nase or sofie). Median growth rates for hybrids were similar and intermediate between those of the parental species. We observed variable life history traits for hybrids and pure forms in the different parts of the hybrid zone. Geometrical analysis of the hybrid fish shape gave evidence of two main morphologies with a link to maternal heritage. Selective mating seemed to be the underlying process which, with mitochondrial heritage, could explain the evolution of the studied hybrid zone. More generally, we showed the importance of studies on hybrid zones and specifically the study of individuals' ecological characteristics, to improve our understanding of speciation.

  4. Fish Vaccine Development and Use to Prevent Streptococcal Diseases

    Science.gov (United States)

    An important pathogen of tilapia, hybrid striped bass and trout raised in intensive aquaculture is Streptococcus sp., a cause of severe economic losses in the fish farming industry. Infected fish experience severe to moderate mortality due to Streptococcus iniae and/or S. agalactiae. The diseased ...

  5. Application of Fluorescence In Situ Hybridization (FISH) Technique for the Detection of Genetic Aberration in Medical Science

    OpenAIRE

    Ratan, Zubair Ahmed; Zaman, Sojib Bin; Mehta, Varshil; Haidere, Mohammad Faisal; Runa, Nusrat Jahan; Akter, Nasrin

    2017-01-01

    Fluorescence in situ hybridization (FISH) is a macromolecule recognition technique, which is considered as a new advent in the field of cytology.?Initially, it was developed as a physical mapping tool to delineate genes within chromosomes. The accuracy and versatility of FISH were subsequently capitalized upon in biological and medical research. This visually appealing technique provides an intermediate degree of resolution between DNA analysis and chromosomal investigations. FISH consists of...

  6. Fluorescence In Situ Hybridization (FISH Signal Analysis Using Automated Generated Projection Images

    Directory of Open Access Journals (Sweden)

    Xingwei Wang

    2012-01-01

    Full Text Available Fluorescence in situ hybridization (FISH tests provide promising molecular imaging biomarkers to more accurately and reliably detect and diagnose cancers and genetic disorders. Since current manual FISH signal analysis is low-efficient and inconsistent, which limits its clinical utility, developing automated FISH image scanning systems and computer-aided detection (CAD schemes has been attracting research interests. To acquire high-resolution FISH images in a multi-spectral scanning mode, a huge amount of image data with the stack of the multiple three-dimensional (3-D image slices is generated from a single specimen. Automated preprocessing these scanned images to eliminate the non-useful and redundant data is important to make the automated FISH tests acceptable in clinical applications. In this study, a dual-detector fluorescence image scanning system was applied to scan four specimen slides with FISH-probed chromosome X. A CAD scheme was developed to detect analyzable interphase cells and map the multiple imaging slices recorded FISH-probed signals into the 2-D projection images. CAD scheme was then applied to each projection image to detect analyzable interphase cells using an adaptive multiple-threshold algorithm, identify FISH-probed signals using a top-hat transform, and compute the ratios between the normal and abnormal cells. To assess CAD performance, the FISH-probed signals were also independently visually detected by an observer. The Kappa coefficients for agreement between CAD and observer ranged from 0.69 to 1.0 in detecting/counting FISH signal spots in four testing samples. The study demonstrated the feasibility of automated FISH signal analysis that applying a CAD scheme to the automated generated 2-D projection images.

  7. Data acquisition. GRAAL experiment. Hybrid reactor experiment. AMS experiment

    International Nuclear Information System (INIS)

    Barancourt, D.; Barbier, G.; Bosson, G.; Bouvier, J.; Gallin-Martel, L.; Meillon, B.; Stassi, P.; Tournier, M.

    1997-01-01

    The main activity of the data acquisition team has consisted in hardware and software developments for the GRAAL experiment with the trigger board, for the 'Reacteurs Hybrides' group with an acquisition board ADCVME8V and for the AMS experiment with the monitoring of the aerogel detector. (authors)

  8. A Hybrid Method for Image Segmentation Based on Artificial Fish Swarm Algorithm and Fuzzy c-Means Clustering

    Directory of Open Access Journals (Sweden)

    Li Ma

    2015-01-01

    Full Text Available Image segmentation plays an important role in medical image processing. Fuzzy c-means (FCM clustering is one of the popular clustering algorithms for medical image segmentation. However, FCM has the problems of depending on initial clustering centers, falling into local optimal solution easily, and sensitivity to noise disturbance. To solve these problems, this paper proposes a hybrid artificial fish swarm algorithm (HAFSA. The proposed algorithm combines artificial fish swarm algorithm (AFSA with FCM whose advantages of global optimization searching and parallel computing ability of AFSA are utilized to find a superior result. Meanwhile, Metropolis criterion and noise reduction mechanism are introduced to AFSA for enhancing the convergence rate and antinoise ability. The artificial grid graph and Magnetic Resonance Imaging (MRI are used in the experiments, and the experimental results show that the proposed algorithm has stronger antinoise ability and higher precision. A number of evaluation indicators also demonstrate that the effect of HAFSA is more excellent than FCM and suppressed FCM (SFCM.

  9. Hybrid striped bass feeds based on fish oil, beef tallow, and eicosapentaenoic acid/docosahexaenoic acid supplements: Insight regarding fish oil sparing and demand for -3 long-chain polyunsaturated fatty acids.

    Science.gov (United States)

    Bowzer, J; Jackson, C; Trushenski, J

    2016-03-01

    Previous research suggests that saturated (SFA) and monounsaturated fatty acid (MUFA) rich lipids, including beef tallow, can make utilization or diet-to-tissue transfer of long-chain polyunsaturated fatty acids (LC-PUFA) more efficient. We hypothesized that using beef tallow as an alternative to fish oil may effectively reduce the LC-PUFA demand of hybrid striped bass × and allow for greater fish oil sparing. Accordingly, we evaluated growth performance and tissue fatty acid profiles of juvenile fish (23.7 ± 0.3 g) fed diets containing menhaden fish oil (considered an ideal source of LC-PUFA for this taxon), beef tallow (BEEF ONLY), or beef tallow amended with purified sources of eicosapentaenoic acid (EPA) and/or docosahexaenoic acid (DHA) to achieve levels corresponding to 50 or 100% of those observed in the FISH ONLY feed. Diets were randomly assigned to quadruplicate tanks of fish ( = 4; 10 fish/tank), and fish were fed assigned diets to apparent satiation once daily for 10 wk. Survival (98-100%) was equivalent among treatments, but weight gain (117-180%), specific growth rate (1.1-1.5% BW/d), feed intake (1.4-1.8% BW/d), thermal growth coefficient (0.50-0.70), and feed conversion ratio (FCR; 1.1-1.4, DM basis) varied. Except for FCR, no differences were observed between the FISH ONLY and BEEF ONLY treatments, but performance was generally numerically superior among fish fed the diets containing beef tallow supplemented with DHA at the 100% or both EPA and DHA at the 50% or 100% level. Tissue fatty acid composition was significantly distorted in favor among fish fed the beef tallow-based feeds; however, profile distortion was most overt in peripheral tissues. Results suggest that beef tallow may be used as a primary lipid source in practical diets for hybrid striped bass, but performance may be improved by supplementation with LC-PUFA, particularly DHA. Furthermore, our results suggest that -3 LC-PUFA requirements reported for hybrid striped bass may not be

  10. When biogeographical provinces collide: Hybridization of reef fishes at the crossroads of marine biogeographical provinces in the Arabian Sea

    KAUST Repository

    DiBattista, Joseph

    2015-04-01

    Aim: Suture zones are areas where closely related species from different biogeographical regions come into contact and interbreed. This concept originated from the study of terrestrial ecosystems but it remains unclear whether a similar phenomenon occurs in the marine environment. Here we investigate a potential suture zone from a previously unknown hybrid hotspot at the Socotra Archipelago (Yemen), located in the Arabian Sea, where fauna from the Red Sea, Gulf of Aden, Arabian Sea, western Indian Ocean and greater Indo-Polynesian Province intersect. Location: Red Sea, Gulf of Aden, Arabian Sea and Indian Ocean. Methods: Putative hybrid reef fish were identified based on intermediate coloration and morphology. Underwater observations and collections were conducted to determine: (1) whether parent species form heterospecific social groups or breeding pairs; (2) the sex and reproductive status of morphologically intermediate individuals; and (3) whether parent species were forming mixed species associations owing to a dearth of conspecific partners. To support hybrid status, morphologically intermediate and parental individuals were genotyped using mitochondrial DNA cytochrome c oxidase subunit I (COI), nuclear recombination-activating gene 2 (RAG2) and the nuclear TMO-4C4 (TMO) gene. Results: We observed putative hybrids involving 14 species from four reef fish families at Socotra. Most cases involved a parental species with a restricted distribution (e.g. Red Sea or Arabian Sea) and a broadly distributed Indo-Pacific species. In most cases, at least one of the parent species was rare at Socotra. Hybrid gene flow was largely unidirectional, and although introgression was rare, we found evidence that some butterflyfish and surgeonfish hybrids were fertile and formed breeding groups with parental species. Main conclusions: The rate of hybrid discovery at Socotra is much greater than that recorded elsewhere in the marine environment and involved both allopatric and

  11. Chromogenic in situ hybridization (CISH) to detect HER2 gene amplification in breast and gastric cancer: comparison with immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH).

    Science.gov (United States)

    Kiyose, Shinichiro; Igarashi, Hisaki; Nagura, Kiyoko; Kamo, Takaharu; Kawane, Kazunori; Mori, Hiroki; Ozawa, Takachika; Maeda, Matsuyoshi; Konno, Keisuke; Hoshino, Hideaki; Konno, Hiroyuki; Ogura, Hiroyuki; Shinmura, Kazuya; Hattori, Naohiko; Sugimura, Haruhiko

    2012-11-01

    The chromogenic in situ hybridization (CISH) assay, designed to detect the amplification of the HER2 gene in formalin-fixed, paraffin-embedded (FFPE) breast cancer (BC) and gastric cancer (GC) tissue specimens, was evaluated in 125 FFPE BC cases and 198 FFPE GC cases for which the HER2 status had been predetermined using immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). In the 125 BC cases and the 198 gastric cases, we found a very good concordance (98.4% and 99.0%, respectively) between CISH and FISH. In particular, we evaluated the polysomy cases, as these cases often have ambiguous treatment options in clinical practice. The polysomy of chromosome 17 was defined as the presence of three or more CEP17 signals in at least 10% of the tumor cells. In the 50 BC cases and 54 GC cases displaying chromosome 17 polysomy, the concordance between FISH and CISH was 98.0% and 98.1%, respectively. These results indicate that CISH could provide an accurate and practical alternative to FISH for the clinical diagnosis of HER2 gene amplification in FFPE BC and FFPE GC samples. © 2012 The Authors. Pathology International © 2012 Japanese Society of Pathology and Wiley Publishing Asia Pty Ltd.

  12. Visualization of mcr mRNA in a methanogen by fluorescence in situ hybridization with an oligonucleotide probe and two-pass tyramide signal amplification (two-pass TSA-FISH).

    Science.gov (United States)

    Kubota, Kengo; Ohashi, Akiyoshi; Imachi, Hiroyuki; Harada, Hideki

    2006-09-01

    Two-pass tyramide signal amplification-fluorescence in situ hybridization (two-pass TSA-FISH) with a horseradish peroxidase (HRP)-labeled oligonucleotide probe was applied to detect prokaryotic mRNA. In this study, mRNA of a key enzyme for methanogenesis, methyl coenzyme M reductase (mcr), in Methanococcus vannielii was targeted. Applicability of mRNA-targeted probes to in situ hybridization was verified by Clone-FISH. It was observed that sensitivity of two-pass TSA-FISH was significantly higher than that of TSA-FISH, which was further increased by the addition of dextran sulphate in TSA working solution. Signals from two-pass TSA-FISH were more reliable compared to the weak, spotty signals yielded by TSA-FISH.

  13. Role of chromogenic in situ hybridization (CISH) in the evaluation of HER2 status in breast carcinoma: comparison with immunohistochemistry and FISH.

    Science.gov (United States)

    Li-Ning-T, Elsa; Ronchetti, Ruben; Torres-Cabala, Carlos; Merino, Maria J

    2005-10-01

    We report our experience with Chromogenic in Situ Hybridization (CISH) for the evaluation of HER2 amplification on 55 cases of formalin-fixed, paraffin-embedded invasive breast carcinomas of different histology. All the results were corrected for chromosome 17 aneusomy and compared with immunohistochemistry (IHC); a subset of cases was compared to FISH. Thirty-one of 32 cases in which FISH and CISH were performed yielded the same results. CISH and IHC showed a good concordance in the 0/1+ and 3+ category, while a poor agreement with weakly protein overexpression was confirmed. Chromosome 17 analysis was necessary in cases with a low number of HER2 gene copies. CISH is a useful tool to evaluate breast cancer HER2 status that can be easily implemented in a laboratory of surgical pathology.

  14. Diagnostic and Prognostic Utility of Fluorescence In situ Hybridization (FISH) Analysis in Acute Myeloid Leukemia.

    Science.gov (United States)

    Gonzales, Patrick R; Mikhail, Fady M

    2017-12-01

    Acute myeloid leukemia (AML) is a hematologic neoplasia consisting of incompletely differentiated hematopoietic cells of the myeloid lineage that proliferate in the bone marrow, blood, and/or other tissues. Clinical implementation of fluorescence in situ hybridization (FISH) in cytogenetic laboratories allows for high-resolution analysis of recurrent structural chromosomal rearrangements specific to AML, especially in AML with normal karyotypes, which comprises approximately 33-50% of AML-positive specimens. Here, we review the use of several FISH probe strategies in the diagnosis of AML. We also review the standards and guidelines currently in place for use by clinical cytogenetic laboratories in the evaluation of AML. Updated standards and guidelines from the WHO, ACMG, and NCCN have further defined clinically significant, recurring cytogenetic anomalies in AML that are detectable by FISH. FISH continues to be a powerful technique in the diagnosis of AML, with higher resolution than conventional cytogenetic analysis, rapid turnaround time, and a considerable diagnostic and prognostic utility.

  15. Growth of monosex hybrid tilapia in the labortory and sewage oxidation ponds

    International Nuclear Information System (INIS)

    Suffern, J.S.; Adams, S.M.; Blaylock, B.G.; Coutant, C.C.; Guthrie, C.A.

    1978-01-01

    Studies were conducted to evaluate the potential of monosex hybrid tilapia (female T. mossambica x male T. hornorum) in waste-heat polyculture systems. The optimum growth temperature for this hybrid was found to be 32 0 C in laboratory experiments. Experiments in sewage pond cage culture in the temperature range of 23 to 33 0 C at stocking densities of approximately 53 fish/m 3 were also conducted. At fish sizes between 5 and 12 cm TL, estimated annual production is approximately 50,000 kg/ha/yr (50,000 lb/acre/yr). Fish in the sewage oxidation ponds grew significantly faster than fish fed trout chow at optimum temperature in the laboratory, even though temperatures in the sewage ponds averaged below the optimum growth temperature. Techniques to accelerate growth rates are being explored. Exposure to gamma radiation (500 rads), known to cause significant increases in channel catfish growth rate, was found to have a similar effect on tilapia. After a 20-week growth period, exposed fish weighed an average of 20% more than controls

  16. Fluctuation localization imaging-based fluorescence in situ hybridization (fliFISH) for accurate detection and counting of RNA copies in single cells

    Energy Technology Data Exchange (ETDEWEB)

    Cui, Yi; Hu, Dehong; Markillie, Lye Meng; Chrisler, William B.; Gaffrey, Matthew J.; Ansong, Charles; Sussel, Lori; Orr, Galya

    2017-10-04

    Quantitative gene expression analysis in intact single cells can be achieved using single molecule- based fluorescence in situ hybridization (smFISH). This approach relies on fluorescence intensity to distinguish between true signals, emitted from an RNA copy hybridized with multiple FISH sub-probes, and background noise. Thus, the precision in smFISH is often compromised by partial or nonspecific binding of sub-probes and tissue autofluorescence, limiting its accuracy. Here we provide an accurate approach for setting quantitative thresholds between true and false signals, which relies on blinking frequencies of photoswitchable dyes. This fluctuation localization imaging-based FISH (fliFISH) uses blinking frequency patterns, emitted from a transcript bound to multiple sub-probes, which are distinct from blinking patterns emitted from partial or nonspecifically bound sub-probes and autofluorescence. Using multicolor fliFISH, we identified radial gene expression patterns in mouse pancreatic islets for insulin, the transcription factor, NKX2-2, and their ratio (Nkx2-2/Ins2). These radial patterns, showing higher values in β cells at the islet core and lower values in peripheral cells, were lost in diabetic mouse islets. In summary, fliFISH provides an accurate, quantitative approach for detecting and counting true RNA copies and rejecting false signals by their distinct blinking frequency patterns, laying the foundation for reliable single-cell transcriptomics.

  17. Fluorescence in situ hybridization techniques (FISH) to detect changes in CYP19a gene expression of Japanese medaka (Oryzias latipes)

    International Nuclear Information System (INIS)

    Park, June-Woo; Tompsett, Amber; Zhang, Xiaowei; Newsted, John L.; Jones, Paul D.; Au, Doris; Kong, Richard; Wu, Rudolf S.S.; Giesy, John P.; Hecker, Markus

    2008-01-01

    The aim of this study was to develop a sensitive in situ hybridization methodology using fluorescence-labeled riboprobes (FISH) that allows for the evaluation of gene expression profiles simultaneously in multiple target tissues of whole fish sections of Japanese medaka (Oryzias latipes). To date FISH methods have been limited in their application due to autofluorescence of tissues, fixatives or other components of the hybridization procedure. An optimized FISH method, based on confocal fluorescence microscopy was developed to reduce the autofluorescence signal. Because of its tissue- and gender-specific expression and relevance in studies of endocrine disruption, gonadal aromatase (CYP19a) was used as a model gene. The in situ hybridization (ISH) system was validated in a test exposure with the aromatase inhibitor fadrozole. The optimized FISH method revealed tissue-specific expression of the CYP19a gene. Furthermore, the assay could differentiate the abundance of CYP19a mRNA among cell types. Expression of CYP19a was primarily associated with early stage oocytes, and expression gradually decreased with increasing maturation. No expression of CYP19a mRNA was observed in other tissues such as brain, liver, or testes. Fadrozole (100 μg/L) caused up-regulation of CYP19a expression, a trend that was confirmed by RT-PCR analysis on excised tissues. In a combination approach with gonad histology, it could be shown that the increase in CYP19a expression as measured by RT-PCR on a whole tissue basis was due to a combination of both increases in numbers of CYP19a-containing cells and an increase in the amount of CYP19a mRNA present in the cells

  18. The Danish Experience with Transferable Fishing Concessions

    DEFF Research Database (Denmark)

    Højrup, Thomas

    2012-01-01

    The task of the concise chapter is to describe the Danish experiences with a basic regulation based upon the principle of Transferable Fisheries Concessions, TFC - at the backgruund of the introduction of Transfreable Quotas in 2003 (pelagic fisheries) and 2007 (demersal fisheries). To do this th...... dependent upon the struggles on the financial markets, replacing sustainable fishing methods with heavy buttom trawling and undermining the future of young people in the fishing industry....

  19. [HER-2 oncogene amplification assessment in invasive breast cancer by dual-color in situ hybridization (dc-CISH): a comparative study with fluorescent in situ hybridization (FISH)].

    Science.gov (United States)

    Akhdar, Abbas; Bronsard, Marc; Lemieux, Renald; Geha, Sameh

    2011-12-01

    The amplification of the gene encoding for the human epidermal growth factor receptor 2 (HER-2 oncogene), located on chromosome 17 (17q21-q22), or the overexpression of this receptor have prognostic and therapeutic implications in invasive breast cancer. An evaluation of the HER-2 status by immunohistochemistry (IHC) is performed on all invasive breast cancer cases. Fluorescent in situ hybridization (FISH) is considered as the gold standard for the detection of HER-2 gene amplification for IHC equivocal cases (score 2+). A more recent in situ hybridization technique, the dual-color chromogenic in situ hybridization (dc-CISH), has been proposed as an alternative to FISH. The aim of this study was to measure the correlation between dc-CISH and FISH for HER-2 oncogene amplification assessment in invasive breast cancer. We built four tissue micro-array (TMA) blocs with 100 breast invasive cancer cases that had been previously tested by IHC for HER-2 detection: 10 score 0 cases, 10 score 3+cases, 39 score 1+and 41 score 2+cases. Both FISH and dc-CISH techniques were applied on all TMA cases as well as on two additional slides serving as controls. Interpretation of dc-CISH was carried out by a pathologist using an optical microscope. For FISH, the interpretation was done by a professional from the medical genetics department using a fluorescent microscope linked to a computer system for image capturing and analysis. The interpretation of the HER-2/CEN-17 ratio for both tests was in accordance with the values of the updated recommendations from the Canadian National Consensus Meeting on HER-2/neu testing in breast cancer and from the ASCO/CAP. Among the 100 cases initially included in the study, eight were excluded from the analysis due to sampling or technical flaws. From the 92 remaining cases, we obtained a concordance of 97.8% (90/92 cases) between the two techniques (Kappa coefficient 0.97, 95% confidence interval). The correlation coefficient (rho) between ratios

  20. 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

    Adhesion of Plasmodium falciparum infected erythrocytes (IE) to human endothelial receptors during malaria infections is mediated by expression of PfEMP1 protein variants encoded by the var genes. The haploid P. falciparum genome harbors approximately 60 different var genes of which only one has...... been believed to be transcribed per cell at a time during the blood stage of the infection. How such mutually exclusive regulation of var gene transcription is achieved is unclear, as is the identification of individual var genes or sub-groups of var genes associated with different receptors...... 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...

  1. Marine Fish Hybridization

    KAUST Repository

    He, Song

    2017-01-01

    for each hybrid offspring in each case, haploweb analysis on diagnostic markers (nuclear and/or mitochondrial) and the DAPC/PCA analysis on microsatellite data were used. By combining the genetic evidences, morphological traits, and ecological observations

  2. PV-Diesel Hybrid SCADA Experiment Network Design

    Science.gov (United States)

    Kalu, Alex; Durand, S.; Emrich, Carol; Ventre, G.; Wilson, W.; Acosta, R.

    1999-01-01

    The essential features of an experimental network for renewable power system satellite based supervisory, control and data acquisition (SCADA) are communication links, controllers, diagnostic equipment and a hybrid power system. Required components for implementing the network consist of two satellite ground stations, to satellite modems, two 486 PCs, two telephone receivers, two telephone modems, two analog telephone lines, one digital telephone line, a hybrid-power system equipped with controller and a satellite spacecraft. In the technology verification experiment (TVE) conducted by Savannah State University and Florida Solar Energy Center, the renewable energy hybrid system is the Apex-1000 Mini-Hybrid which is equipped with NGC3188 for user interface and remote control and the NGC2010 for monitoring and basic control tasks. This power system is connected to a satellite modem via a smart interface, RS232. Commands are sent to the power system control unit through a control PC designed as PC1. PC1 is thus connected to a satellite model through RS232. A second PC, designated PC2, the diagnostic PC is connected to both satellite modems via separate analog telephone lines for checking modems'health. PC2 is also connected to PC1 via a telephone line. Due to the unavailability of a second ground station for the ACTS, one ground station is used to serve both the sending and receiving functions in this experiment. Signal is sent from the control PC to the Hybrid system at a frequency f(sub 1), different from f(sub 2), the signal from the hybrid system to the control PC. f(sub l) and f(sub 2) are sufficiently separated to avoid interference.

  3. Fluorescence in situ hybridization (CARD-FISH) of microorganisms in hydrocarbon contaminated aquifer sediment samples.

    Science.gov (United States)

    Tischer, Karolin; Zeder, Michael; Klug, Rebecca; Pernthaler, Jakob; Schattenhofer, Martha; Harms, Hauke; Wendeberg, Annelie

    2012-12-01

    Groundwater ecosystems are the most important sources of drinking water worldwide but they are threatened by contamination and overexploitation. Petroleum spills account for the most common source of contamination and the high carbon load results in anoxia and steep geochemical gradients. Microbes play a major role in the transformation of petroleum hydrocarbons into less toxic substances. To investigate microbial populations at the single cell level, fluorescence in situ hybridization (FISH) is now a well-established technique. Recently, however, catalyzed reporter deposition (CARD)-FISH has been introduced for the detection of microbes from oligotrophic environments. Nevertheless, petroleum contaminated aquifers present a worst case scenario for FISH techniques due to the combination of high background fluorescence of hydrocarbons and the presence of small microbial cells caused by the low turnover rates characteristic of groundwater ecosystems. It is therefore not surprising that studies of microorganisms from such sites are mostly based on cultivation techniques, fingerprinting, and amplicon sequencing. However, to reveal the population dynamics and interspecies relationships of the key participants of contaminant degradation, FISH is an indispensable tool. In this study, a protocol for FISH was developed in combination with cell quantification using an automated counting microscope. The protocol includes the separation and purification of microbial cells from sediment particles, cell permeabilization and, finally, CARD-FISH in a microwave oven. As a proof of principle, the distribution of Archaea and Bacteria was shown in 60 sediment samples taken across the contaminant plume of an aquifer (Leuna, Germany), which has been heavily contaminated with several ten-thousand tonnes of petroleum hydrocarbons since World War II. Copyright © 2012 Elsevier GmbH. All rights reserved.

  4. Utility and impact of early t(15;17) identification by Fluorescence In Situ Hybridization (FISH) in clinical decision making for patients in Acute Promyelocytic Leukemia (APL).

    Science.gov (United States)

    Kolhe, R; Mangaonkar, A; Mansour, J; Clemmons, A; Shaw, J; Dupont, B; Walczak, L; Mondal, A; Rojiani, A; Jillella, A; Kota, V

    2015-08-01

    Acute Promyelocytic Leukemia (APL) is a curable malignancy with studies showing above 90% survival. However, population-based studies looking at survival suggest that approximately 30% of patients with APL die during induction. Early demonstration of t(15;17) will lead to accurate decision making regarding treatment. The aim of this project was to validate earlier time frames for the Abbott Molecular Vysis LSI promyelocytic leukemia (PML)/ retinoic acid receptor alpha (RARA) fluorescence in situ hybridization (FISH) probe (ASR 6-16 h). Twenty patients (15 APL cases and five non-APL cases) were selected for validating various hybridization times for the FISH probe. Expected normal signal pattern was two red and two green signals (2R2G), and the most common expected abnormal signal pattern was two fusion (yellow) signals, one red and one green (2F1R1G) and/or one fusion, one red and one green (1F1R1G). The specificity of the probe ranged from 84% at 2 h, 86% at 4 h, 84% at 6 h, and 87% for overnight hybridization. The sensitivity increased from 79% at 2 h, 80% at 4 h, 81% at 6 h to 87% for overnight hybridization. Based on the validation studies, we recommend reading of FISH results at the 4-h incubation mark for a preliminary diagnosis and confirmation with overnight hybridization. © 2015 John Wiley & Sons Ltd.

  5. Fluorescent in situ hybridization (FISH as a diagnostic tool for Williams-Beuren syndrome

    Directory of Open Access Journals (Sweden)

    Deise Helena de Souza

    2007-01-01

    Full Text Available Fluorescent in situ hybridization (FISH with commercial probes covering the elastin gene (ELN was used to determine the frequency of the 7q11.23 deletion in 18 children clinically diagnosed with Williams-Beuren syndrome (WBS. A de novo deletion was detected in 15 of the children (83%. Diagnostic investigation for WBS started late in childhood (median = 5.8 years. All the children showed facial features typical of the syndrome, mental retardation and developmental delay. Over-friendliness was observed in the majority of cases. Clinodactyly of the 5th finger (n = 13, cardiovascular disease (n = 9, loquacity (n = 9, low birthweight (n = 8, and failure to thrive (n = 9 were observed only in those children with the deletion. Respiratory problems (n = 9, though not previously reported in the literature, was a common finding in the group studied. Our results confirmed that FISH is useful in identifying 7q11.23 deletions in cases of WBS. Clinical manifestations were more evident in the deletion-positive children.

  6. Hybrid Wars: Israel’s experience

    Directory of Open Access Journals (Sweden)

    E. N. Grachikov

    2015-01-01

    Full Text Available The article attempts to summarize the experience of Israel in the fight against non-traditional threats, which escalated into a hybrid war. Feature of these wars lies in the fact that they are usually conducted between Western and non-Western countries in the border and poorly controlled space. West opponents favor alliances and associations of non-Western countries and non-state actors. In this case, to justify the war, the norms of international law are used.

  7. Fluorescence in situ hybridization on formalin-fixed and paraffin-embedded tissue

    DEFF Research Database (Denmark)

    Laub Petersen, Bodil; Zeuthen, Mette Christa; Pedersen, Sanni

    2004-01-01

    Fluorescence in situ hybridization (FISH) is widely used to study numerical and structural genetic abnormalities in both metaphase and interphase cells. The technique is based on the hybridization of labeled probes to complementary sequences in the DNA or RNA of the cells. Interphase FISH is most...... in time lapse between removal of tissue and fixation, duration of fixation, enzymatic pretreatment, hybridization conditions, and posthybridization washing conditions are important factors in the hybridization. In this study, we have listed the results of a systematic approach to improve FISH on isolated...

  8. Growth of monosex hybrid tilapia in the labortory and sewage oxidation ponds. [Effects of water temperature, nutrient level, and. gamma. rays on growth

    Energy Technology Data Exchange (ETDEWEB)

    Suffern, J.S.; Adams, S.M.; Blaylock, B.G.; Coutant, C.C.; Guthrie, C.A.

    1978-01-01

    Studies were conducted to evaluate the potential of monosex hybrid tilapia (female T. mossambica x male T. hornorum) in waste-heat polyculture systems. The optimum growth temperature for this hybrid was found to be 32/sup 0/C in laboratory experiments. Experiments in sewage pond cage culture in the temperature range of 23 to 33/sup 0/C at stocking densities of approximately 53 fish/m/sup 3/ were also conducted. At fish sizes between 5 and 12 cm TL, estimated annual production is approximately 50,000 kg/ha/yr (50,000 lb/acre/yr). Fish in the sewage oxidation ponds grew significantly faster than fish fed trout chow at optimum temperature in the laboratory, even though temperatures in the sewage ponds averaged below the optimum growth temperature. Techniques to accelerate growth rates are being explored. Exposure to gamma radiation (500 rads), known to cause significant increases in channel catfish growth rate, was found to have a similar effect on tilapia. After a 20-week growth period, exposed fish weighed an average of 20% more than controls.

  9. Identification of the centromeric repeat in the threespine stickleback fish (Gasterosteus aculeatus).

    Science.gov (United States)

    Cech, Jennifer N; Peichel, Catherine L

    2015-12-01

    Centromere sequences exist as gaps in many genome assemblies due to their repetitive nature. Here we take an unbiased approach utilizing centromere protein A (CENP-A) chomatin immunoprecipitation followed by high-throughput sequencing to identify the centromeric repeat sequence in the threespine stickleback fish (Gasterosteus aculeatus). A 186-bp, AT-rich repeat was validated as centromeric using both fluorescence in situ hybridization (FISH) and immunofluorescence combined with FISH (IF-FISH) on interphase nuclei and metaphase spreads. This repeat hybridizes strongly to the centromere on all chromosomes, with the exception of weak hybridization to the Y chromosome. Together, our work provides the first validated sequence information for the threespine stickleback centromere.

  10. Evaluation of HER-2/neu status in breast cancer specimens using immunohistochemistry (IHC) & fluorescence in-situ hybridization (FISH) assay.

    Science.gov (United States)

    Goud, Kalal Iravathy; Dayakar, Seetha; Vijayalaxmi, Kolanupaka; Babu, Saidam Jangu; Reddy, P Vijay Anand

    2012-03-01

    Fluorescence in situ hybridization (FISH) is increasingly being recognized as the most accurate and predictive test for HER 2/neu gene amplification and response to therapy in breast cancer. In the present study we investigated HER-2/neu gene amplification by FISH in breast carcinoma tissue specimens and compared the results with that of immunohistochemical (IHC) analysis. A total of 90 breast carcinoma tissue samples were used for immunohistochemical (IHC) and FISH analysis. IHC was performed by using mouse monoclonal antibody to the intracellular domain of HER-2/neu protein. Each slide was scored in a blinded fashion by two pathologists according to the manufacturer's recommended criteria. FISH analysis was performed on paraffin embedded breast tumour tissue sections. The polysomy for centromere 17 (Spec green signal) was read as green signals less than 4 as moderate polysomy, and more than 4 as highly polysomy. Thirty of the 90 patients had negative results by IHC and FISH. Of the 28 patients with the score of 2+ by IHC, 20 were FISH positive for HER-2/neu gene amplification, three were FISH negative and five patients showed equivocal (1.8-2.2) results by FISH. These five cases were retested for IHC and FISH on different paraffin embedded tissue blocks, and all five were found positive for HER-2/neu gene amplification. Twenty five patients with the score of 3+ by IHC were FISH positive for HER-2/neu gene amplification (>2.2). Seven cases with the score of 3+ by IHC were FISH negative for HER-2/neu gene amplification (>2.2), and showed polysomy of chromosome number 17 high polysomy > 4. Our results indicated that HER-2/neu status by FISH should be performed in all cases of breast tumour with a 2+ score by IHC. Cases demonstrating a 3+ score by IHC may be subjected to FISH to rule out polysomy of chromosome 17 which could be falsely interpreted as HER-2/neu overexpression by IHC analysis. There is also a need for establishing a clinically validated cut-off value

  11. Genetic identification of F1 and post-F1 serrasalmid juvenile hybrids in Brazilian aquaculture.

    Directory of Open Access Journals (Sweden)

    Diogo Teruo Hashimoto

    Full Text Available Juvenile fish trade monitoring is an important task on Brazilian fish farms. However, the identification of juvenile fish through morphological analysis is not feasible, particularly between interspecific hybrids and pure species individuals, making the monitoring of these individuals difficult. Hybrids can be erroneously identified as pure species in breeding facilities, which might reduce production on farms and negatively affect native populations due to escapes or stocking practices. In the present study, we used a multi-approach analysis (molecular and cytogenetic markers to identify juveniles of three serrasalmid species (Colossoma macropomum, Piaractus mesopotamicus and Piaractus brachypomus and their hybrids in different stocks purchased from three seed producers in Brazil. The main findings of this study were the detection of intergenus backcrossing between the hybrid ♀ patinga (P. mesopotamicus×P. brachypomus×♂ C. macropomum and the occurrence of one hybrid triploid individual. This atypical specimen might result from automixis, a mechanism that produces unreduced gametes in some organisms. Moreover, molecular identification indicated that hybrid individuals are traded as pure species or other types of interspecific hybrids, particularly post-F1 individuals. These results show that serrasalmid fish genomes exhibit high genetic heterogeneity, and multi-approach methods and regulators could improve the surveillance of the production and trade of fish species and their hybrids, thereby facilitating the sustainable development of fish farming.

  12. Patterns of parasite distribution in the hybrids of non-congeneric cyprinid fish species: is asymmetry in parasite infection the result of limited coadaptation?

    Czech Academy of Sciences Publication Activity Database

    Krasnovyd, V.; Vetešník, Lukáš; Gettová, L.; Civáňová, K.; Šimková, A.

    2017-01-01

    Roč. 47, č. 8 (2017), s. 471-483 ISSN 0020-7519 R&D Projects: GA ČR GAP505/12/0375 Institutional support: RVO:68081766 Keywords : Interspecific hybrids * Cyprinid fish * Parasite communities * Host specificity * Maternal ancestry Subject RIV: EG - Zoology OBOR OECD: Parasitology Impact factor: 3.730, year: 2016

  13. Production of F1 interspecies hybrid offspring with cryopreserved sperm from a live-bearing fish, the swordtail Xiphophorus helleri.

    Science.gov (United States)

    Yang, Huiping; Hazlewood, Leona; Heater, Sheila J; Guerrero, Paula A; Walter, Ronald B; Tiersch, Terrence R

    2007-03-01

    Despite study of sperm cryopreservation in more than 200 fish species, production of broods from cryopreserved sperm in live-bearing fish has not been demonstrated. This has not been due to a lack of effort, but instead is a result of the unique morphology, biology, and biochemistry of reproduction in viviparous fishes. For example, sperm of Xiphophorus helleri have a cylindrical nucleus, can swim for days after being activated, have glycolytic capabilities, and can reside in the female reproduction tract for months before fertilization. These traits are not found in fishes with external fertilization. The long-standing research use of the genus Xiphophorus has led to development of over 60 pedigreed lines among the 26 species maintained around the world. These species and lines serve as contemporary models in medical research, although they must be maintained as live populations. Previous attempts at establishing sperm cryopreservation protocols for Xiphophorus have not produced live young. To address this we have been studying the parameters surrounding cryobiology of Xiphophorus sperm and applying this information to an improved understanding of internal fertilization and reproduction. Here we report the first successful fertilization and offspring production by cryopreserved sperm in any live-bearing fish. This claim is supported by our use of artificial insemination between two species that yield distinct hybrid offspring to verify paternity via cryopreserved sperm. We provide a practical approach for preservation of valuable genetic resources from live-bearing fish species, a group that is rapidly being lost due to destruction of native habitats.

  14. Spacecraft Hybrid (Mixed-Actuator) Attitude Control Experiences on NASA Science Missions

    Science.gov (United States)

    Dennehy, Cornelius J.

    2014-01-01

    There is a heightened interest within NASA for the design, development, and flight implementation of mixed-actuator hybrid attitude control systems for science spacecraft that have less than three functional reaction wheel actuators. This interest is driven by a number of recent reaction wheel failures on aging, but what could be still scientifically productive, NASA spacecraft if a successful hybrid attitude control mode can be implemented. Over the years, hybrid (mixed-actuator) control has been employed for contingency attitude control purposes on several NASA science mission spacecraft. This paper provides a historical perspective of NASA's previous engineering work on spacecraft mixed-actuator hybrid control approaches. An update of the current situation will also be provided emphasizing why NASA is now so interested in hybrid control. The results of the NASA Spacecraft Hybrid Attitude Control Workshop, held in April of 2013, will be highlighted. In particular, the lessons learned captured from that workshop will be shared in this paper. An update on the most recent experiences with hybrid control on the Kepler spacecraft will also be provided. This paper will close with some future considerations for hybrid spacecraft control.

  15. 5-bp Classical Satellite DNA Loci from Chromosome-1 Instability in Cervical Neoplasia Detected by DNA Breakage Detection/Fluorescence in Situ Hybridization (DBD-FISH

    Directory of Open Access Journals (Sweden)

    Jaime Gosálvez

    2013-02-01

    Full Text Available We aimed to evaluate the association between the progressive stages of cervical neoplasia and DNA damage in 5-bp classical satellite DNA sequences from chromosome-1 in cervical epithelium and in peripheral blood lymphocytes using DNA breakage detection/fluorescence in situ hybridization (DBD-FISH. A hospital-based unmatched case-control study was conducted in 2011 with a sample of 30 women grouped according to disease stage and selected according to histological diagnosis; 10 with low-grade squamous intraepithelial lesions (LG-SIL, 10 with high-grade SIL (HG-SIL, and 10 with no cervical lesions, from the Unidad Medica de Alta Especialidad of The Mexican Social Security Institute, IMSS, Mexico. Specific chromosome damage levels in 5-bp classical satellite DNA sequences from chromosome-1 were evaluated in cervical epithelium and peripheral blood lymphocytes using the DBD-FISH technique. Whole-genome DNA hybridization was used as a reference for the level of damage. Results of Kruskal-Wallis test showed a significant increase according to neoplastic development in both tissues. The instability of 5-bp classical satellite DNA sequences from chromosome-1 was evidenced using chromosome-orientation FISH. In conclusion, we suggest that the progression to malignant transformation involves an increase in the instability of 5-bp classical satellite DNA sequences from chromosome-1.

  16. 5-bp Classical Satellite DNA Loci from Chromosome-1 Instability in Cervical Neoplasia Detected by DNA Breakage Detection/Fluorescence in Situ Hybridization (DBD-FISH).

    Science.gov (United States)

    Cortés-Gutiérrez, Elva I; Ortíz-Hernández, Brenda L; Dávila-Rodríguez, Martha I; Cerda-Flores, Ricardo M; Fernández, José Luis; López-Fernández, Carmen; Gosálvez, Jaime

    2013-02-19

    We aimed to evaluate the association between the progressive stages of cervical neoplasia and DNA damage in 5-bp classical satellite DNA sequences from chromosome-1 in cervical epithelium and in peripheral blood lymphocytes using DNA breakage detection/fluorescence in situ hybridization (DBD-FISH). A hospital-based unmatched case-control study was conducted in 2011 with a sample of 30 women grouped according to disease stage and selected according to histological diagnosis; 10 with low-grade squamous intraepithelial lesions (LG-SIL), 10 with high-grade SIL (HG-SIL), and 10 with no cervical lesions, from the Unidad Medica de Alta Especialidad of The Mexican Social Security Institute, IMSS, Mexico. Specific chromosome damage levels in 5-bp classical satellite DNA sequences from chromosome-1 were evaluated in cervical epithelium and peripheral blood lymphocytes using the DBD-FISH technique. Whole-genome DNA hybridization was used as a reference for the level of damage. Results of Kruskal-Wallis test showed a significant increase according to neoplastic development in both tissues. The instability of 5-bp classical satellite DNA sequences from chromosome-1 was evidenced using chromosome-orientation FISH. In conclusion, we suggest that the progression to malignant transformation involves an increase in the instability of 5-bp classical satellite DNA sequences from chromosome-1.

  17. Considerations in the use of fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy to characterize rumen methanogens and define their spatial distributions.

    Science.gov (United States)

    Valle, Edith R; Henderson, Gemma; Janssen, Peter H; Cox, Faith; Alexander, Trevor W; McAllister, Tim A

    2015-06-01

    In this study, methanogen-specific coenzyme F420 autofluorescence and confocal laser scanning microscopy were used to identify rumen methanogens and define their spatial distribution in free-living, biofilm-, and protozoa-associated microenvironments. Fluorescence in situ hybridization (FISH) with temperature-controlled hybridization was used in an attempt to describe methanogen diversity. A heat pretreatment (65 °C, 1 h) was found to be a noninvasive method to increase probe access to methanogen RNA targets. Despite efforts to optimize FISH, 16S rRNA methanogen-specific probes, including Arch915, bound to some cells that lacked F420, possibly identifying uncharacterized Methanomassiliicoccales or reflecting nonspecific binding to other members of the rumen bacterial community. A probe targeting RNA from the methanogenesis-specific methyl coenzyme M reductase (mcr) gene was shown to detect cultured Methanosarcina cells with signal intensities comparable to those of 16S rRNA probes. However, the probe failed to hybridize with the majority of F420-emitting rumen methanogens, possibly because of differences in cell wall permeability among methanogen species. Methanogens were shown to integrate into microbial biofilms and to exist as ecto- and endosymbionts with rumen protozoa. Characterizing rumen methanogens and defining their spatial distribution may provide insight into mitigation strategies for ruminal methanogenesis.

  18. Human chromosome-specific changes in a human-hamster hybrid cell line (AL) assessed by fluorescent in situ hybridization (fish)

    International Nuclear Information System (INIS)

    Geard, Charles R.; Jenkins, Gloria

    1995-01-01

    Purpose: To quantitatively assess all gamma-ray induced chromosomal changes confined to one human chromosome using fluorescence microscopy and in situ hybridization with a fluorescently labeled human chromosome specific nucleic acid probe. Methods and Materials: Synchronized human-hamster hybrid cells containing human chromosome 11 were obtained by a modified mitotic shake-off procedure. G1 phase cells (> 95%) were irradiated with 137 Cs gamma rays (0, 0.5, 1.0, 1.5, 2.0, 4.0, 6.0, 8.0, and 10.0 Gy) at a dose rate of 1.1 Gy/min and mitotic cells collected 16-20 h later; chromosomal spreads were prepared, denatured, and hybridized with a fluorescein-tagged nucleic acid probe against total human DNA. Chromosomes were examined by fluorescence microscopy and all categories of change involving the human chromosome 11 as target, recorded. Results: Overall, of the 3104 human-hamster hybrid cells examined, 82.1% were euploid, of which 88.6% contained one copy of human chromosome 11, 6.2% contained two copies, and 5.2% contained 0 copies. This is compatible with mitotic nondisjunction in a small fraction of cells. Of the remaining 17.9% of cells, 85.2% were tetraploid cells with two copies of human chromosome 11. For all aberrations involving human chromosome 11 there was a linear relationship between yield and absorbed dose of 0.1 aberrations per chromosome per Gy. The yield of dicentrics, translocations, and terminal deletions that involve one lesion on the human chromosome was linear, while the yield of interstitial deletions that arise from two interacting lesions on the human chromosome was curvilinear. The frequencies of dicentrics and translocations were about equal, while there was a high (40-60%) incidence of incomplete exchanges between human and hamster chromosomes. Conclusions: Fluorescent in situ hybridization (FISH) procedures allow for the efficient detection of a broad range of induced changes in target chromosomes. Symmetrical exchanges induced in G1

  19. Common Fluorescence In Situ Hybridization Applications in Cytology.

    Science.gov (United States)

    Savic, Spasenija; Bubendorf, Lukas

    2016-12-01

    - Fluorescence in situ hybridization (FISH) is a well-established method for detection of genomic aberrations in diagnostic, prognostic, and predictive marker testing. - To review common applications of FISH in cytology. - The published literature was reviewed. - Cytology is particularly well suited for all kinds of FISH applications, which is highlighted in respiratory tract cytology with an increasing demand for predictive FISH testing in lung cancer. Fluorescence in situ hybridization is the gold standard for detection of predictive anaplastic lymphoma kinase gene (ALK) rearrangements, and the same evaluation criteria as in histology apply to cytology. Several other gene rearrangements, including ROS proto-oncogene 1 receptor tyrosine kinase (ROS1), are becoming clinically important and share the same underlining cytogenetic mechanisms with ALK. MET amplification is one of the most common mechanisms of acquired resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors and can be targeted by crizotinib. As genomic aberrations are a hallmark of malignant cells, FISH is a valuable objective ancillary diagnostic tool. In urinary tract cytology, atypical urothelial cells equivocal for malignancy are a common diagnostic dilemma and multitarget FISH can help clarify such cells. Diagnosis of malignant mesothelioma remains one of the most challenging fields in effusion cytology, and ancillary FISH is useful in establishing the diagnosis. Fluorescence in situ hybridization is a morphology-based technique, and the prerequisite for reliable FISH results is a targeted evaluation of the cells in question (eg, cancer or atypical cells). Cytopathologists and cytotechnicians should therefore be involved in molecular testing in order to select the best material and to provide their morphologic expertise.

  20. The campaign to DNA barcode all fishes, FISH-BOL.

    Science.gov (United States)

    Ward, R D; Hanner, R; Hebert, P D N

    2009-02-01

    FISH-BOL, the Fish Barcode of Life campaign, is an international research collaboration that is assembling a standardized reference DNA sequence library for all fishes. Analysis is targeting a 648 base pair region of the mitochondrial cytochrome c oxidase I (COI) gene. More than 5000 species have already been DNA barcoded, with an average of five specimens per species, typically vouchers with authoritative identifications. The barcode sequence from any fish, fillet, fin, egg or larva can be matched against these reference sequences using BOLD; the Barcode of Life Data System (http://www.barcodinglife.org). The benefits of barcoding fishes include facilitating species identification, highlighting cases of range expansion for known species, flagging previously overlooked species and enabling identifications where traditional methods cannot be applied. Results thus far indicate that barcodes separate c. 98 and 93% of already described marine and freshwater fish species, respectively. Several specimens with divergent barcode sequences have been confirmed by integrative taxonomic analysis as new species. Past concerns in relation to the use of fish barcoding for species discrimination are discussed. These include hybridization, recent radiations, regional differentiation in barcode sequences and nuclear copies of the barcode region. However, current results indicate these issues are of little concern for the great majority of specimens.

  1. Determination of the ruminant origin of bone particles using fluorescence in situ hybridization (FISH).

    Science.gov (United States)

    Lecrenier, M C; Ledoux, Q; Berben, G; Fumière, O; Saegerman, C; Baeten, V; Veys, P

    2014-07-17

    Molecular biology techniques such as PCR constitute powerful tools for the determination of the taxonomic origin of bones. DNA degradation and contamination by exogenous DNA, however, jeopardise bone identification. Despite the vast array of techniques used to decontaminate bone fragments, the isolation and determination of bone DNA content are still problematic. Within the framework of the eradication of transmissible spongiform encephalopathies (including BSE, commonly known as "mad cow disease"), a fluorescence in situ hybridization (FISH) protocol was developed. Results from the described study showed that this method can be applied directly to bones without a demineralisation step and that it allows the identification of bovine and ruminant bones even after severe processing. The results also showed that the method is independent of exogenous contamination and that it is therefore entirely appropriate for this application.

  2. Heavy ion-induced chromosomal aberrations analyzed by fluorescence in situ hybridization

    International Nuclear Information System (INIS)

    Durante, M.; Gialanella, G.; Grossi, G.; Pugliese, M.; Cella, L.; Greco, O.; George, K.; Yang, T.C.

    1997-01-01

    We have investigated the effectiveness of heavy ions in the induction of chromosomal aberrations in mammalian cells by the recent technique of fluorescence in situ hybridization (FISH) with whole-chromosome probes. FISH-painting was used both in metaphase and interphase (prematurely condensed) chromosomes. The purpose of our experiments was to address the following problems: (a) the ratio of different types of aberrations as a function of radiation quality (search for biomarkers); (b) the ratio between aberrations scored in interphase and metaphase as a function of radiation quality (role of apoptosis); (c) differences between cytogenetic effects produced by different ions at the same LET (role of track structure). (orig./MG)

  3. Heavy ion-induced chromosomal aberrations analyzed by fluorescence in situ hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Durante, M; Gialanella, G; Grossi, G; Pugliese, M [Univ. ` ` Federico II` ` , Naples (Italy). Dept. of Physics; [INFN, Naples (Italy); Cella, L; Greco, O [Univ. ` ` Federico II` ` , Naples (Italy). Dept. of Physics; Furusawa, Y [NIRS, Chiba (Japan); George, K; Yang, T C [NASA Lyndon B. Johnson Space Center, Houston, TX (United States)

    1997-09-01

    We have investigated the effectiveness of heavy ions in the induction of chromosomal aberrations in mammalian cells by the recent technique of fluorescence in situ hybridization (FISH) with whole-chromosome probes. FISH-painting was used both in metaphase and interphase (prematurely condensed) chromosomes. The purpose of our experiments was to address the following problems: (a) the ratio of different types of aberrations as a function of radiation quality (search for biomarkers); (b) the ratio between aberrations scored in interphase and metaphase as a function of radiation quality (role of apoptosis); (c) differences between cytogenetic effects produced by different ions at the same LET (role of track structure). (orig./MG)

  4. Lower hybrid heating experiments in tokamaks: an overview

    International Nuclear Information System (INIS)

    Porkolab, M.

    1985-10-01

    Lower hybrid wave propagation theory relevant to heating fusion grade plasmas (tokamaks) is reviewed. A brief discussion of accessibility, absorption, and toroidal ray propagation is given. The main part of the paper reviews recent results in heating experiments on tokamaks. Both electron and ion heating regimes will be discussed. The prospects of heating to high temperatures in reactor grade plasmas will be evaluated

  5. Rapid molecular cytogenetic analysis of X-chromosomal microdeletions: Fluorescence in situ hybridization (FISH) for complex glycerol kinase deficiency

    Energy Technology Data Exchange (ETDEWEB)

    Worley, K.C.; Lindsay, E.A.; McCabe, E.R.B. [Baylor College of Medicine, Houston, TX (United States)] [and others

    1995-07-17

    Diagnosis of X-chromosomal microdeletions has relied upon the traditional methods of Southern blotting and DNA amplification, with carrier identification requiring time-consuming and unreliable dosage calculations. In this report, we describe rapid molecular cytogenetic identification of deleted DNA in affected males with the Xp21 contiguous gene syndrome (complex glycerol kinase deficiency, CGKD) and female carriers for this disorder. CGKD deletions involve the genes for glycerol kinase, Duchenne muscular dystrophy, and/or adrenal hypoplasia congenita. We report an improved method for diagnosis of deletions in individuals with CGKD and for identification of female carriers within their families using fluorescence in situ hybridization (FISH) with a cosmid marker (cosmid 35) within the glycerol kinase gene. When used in combination with an Xq control probe, affected males demonstrate a single signal from the control probe, while female carriers demonstrate a normal chromosome with two signals, as well as a deleted chromosome with a single signal from the control probe. FISH analysis for CGKD provides the advantages of speed and accuracy for evaluation of submicroscopic X-chromosome deletions, particularly in identification of female carriers. In addition to improving carrier evaluation, FISH will make prenatal diagnosis of CGKD more readily available. 17 refs., 2 figs.

  6. Asexual Reproduction Does Not Apparently Increase the Rate of Chromosomal Evolution: Karyotype Stability in Diploid and Triploid Clonal Hybrid Fish (Cobitis, Cypriniformes, Teleostei)

    Czech Academy of Sciences Publication Activity Database

    Majtánová, Zuzana; Choleva, Lukáš; Symonová, Radka; Ráb, Petr; Kotusz, J.; Pekárik, L.; Janko, Karel

    2016-01-01

    Roč. 11, č. 1 (2016), e0146872-e0146872 E-ISSN 1932-6203 R&D Projects: GA ČR GAP506/10/1155; GA ČR GPP506/12/P857; GA ČR GA13-12580S Institutional support: RVO:67985904 Keywords : in-situ hybridization * fresh water fish * unisexual salamanders Subject RIV: EG - Zoology Impact factor: 2.806, year: 2016

  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. Summer yield of fish in polyculture in Transkei, South Africa, using ...

    African Journals Online (AJOL)

    Keywords Aquaculture, fish prOduction, polyculture, integrated farming ... *To whom all correspondence should be addressed. Introduction. The use of animal wastes in fish production systems, has been widely practised for centuries in the Far Eastern countries ..... water without problems in fish-pond management. Hybrids.

  9. Design and analysis methods for fish survival experiments based on release-recapture

    National Research Council Canada - National Science Library

    Burnham, Kenneth P

    1987-01-01

    .... The application of the methods developed here is more general, however, as it includes experiments to estimate survival of fish as they pass over spillways or through bypass systems and several dams...

  10. Transit experience with hydrogen fueled hybrid electric buses

    International Nuclear Information System (INIS)

    Scott, P.B.; Mazaika, D.M.; Levin, J.; Edwards, T.

    2006-01-01

    Both AC Transit and SunLine Transit operate hybrid electric hydrogen fueled buses in their transit service. ACT presently operates three fuel cell buses in daily revenue service, and SunLine operates a fuel cell bus and a HHICE (Hybrid Hydrogen Internal Combustion Engine) bus. All these buses use similar electric drive train and electric accessories, although the detailed design differs notably between the fuel cell and the hybrid ICE buses. The fuel cell buses use a 120kW UTC fuel cell and a Van Hool Chassis, whereas the HHICE bus uses a turbocharged Ford engine which is capable of 140kW generator output in a New Flyer Chassis. The HHICE bus was the first in service, and has been subjected to both winter testing in Manitoba, Canada and summer testing in the Palm Springs, CA region. The winter testing included passenger sampling using questionnaires to ascertain passenger response. The fuel cell buses were introduced to service at the start of 2006. All five buses are in daily revenue service use. The paper will describe the buses and the experience of the transit properties in operating the buses. (author)

  11. Household preferences of hybrid home heating systems – A choice experiment application

    International Nuclear Information System (INIS)

    Ruokamo, Enni

    2016-01-01

    The residential heating sector presents considerable energy savings potential, as numerous heating solutions for reducing electricity consumption and utilizing renewable energy sources are available in the market. The aim of this paper is to examine determinants of household heating system choices and to use this information for policy planning purposes. This paper investigates residential homeowner attitudes regarding innovative hybrid home heating systems (HHHS) with choice experiment. Heating system scenarios are designed to represent the most relevant primary and supplementary heating alternatives currently available in Finland. The choice sets include six main heating alternatives (district heat, solid wood, wood pellet, electric storage heating, ground heat pump and exhaust air heat pump) that are described by five attributes (supplementary heating systems, investment costs, operating costs, comfort of use and environmental friendliness). The results imply that HHHSs generally appear to be accepted among households; however, several factors affect perceptions of these technologies. The results reveal differing household attitudes toward the main heating alternatives and show that such views are affected by socio-demographic characteristics (age, living environment, education, etc.). The results suggest that households view supplementary heating systems (especially solar-based) favorably. The other attributes studied also play a significant role in decision making. - Highlights: •Study of hybrid heating where supplementary and main heating systems are combined. •Choice experiment is applied to study the determinants of hybrid heating adoption. •Hybrid heating appears to be generally accepted among households. •Households exhibit differing attitudes toward hybrid heating. •Policy makers should not underestimate the potential of hybrid heating.

  12. 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

    target site. Finally, the RCA product inside the cells was detected by standard fluorescence in situ hybridization (FISH). The optimized protocol showed high specificity and signal-to-noise ratio but low detection frequency (up to 15% for single-copy genes and up to 43% for the multi-copy 16S rRNA 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.......). Nevertheless, multiple genes (nirS and nosZ; nirS and the 16S rRNA gene) could be detected simultaneously in P. stutzeri. Environmental application of in situ RCA-FISH was demonstrated on activated sludge by the differential detection of two types of nirS-defined denitrifiers; one of them was identified...

  13. Comparison of Chromogenic In Situ Hybridization and Fluorescence In Situ Hybridization for the Evaluation of MDM2 Amplification in Adipocytic Tumors.

    Science.gov (United States)

    Mardekian, Stacey K; Solomides, Charalambos C; Gong, Jerald Z; Peiper, Stephen C; Wang, Zi-Xuan; Bajaj, Renu

    2015-11-01

    Atypical lipomatous tumor/well-differentiated liposarcoma (ALT-WDLPS) and dedifferentiated liposarcoma (DDLPS) are characterized cytogenetically by a 12q13-15 amplification involving the mouse double minute 2 (MDM2) oncogene. Fluorescence in situ hybridization (FISH) is used frequently to detect this amplification and aid with the diagnosis of these entities, which is difficult by morphology alone. Recently, bright-field in situ hybridization techniques such as chromogenic in situ hybridization (CISH) have been introduced for the determination of MDM2 amplification status. The present study compared the results of FISH and CISH for detecting MDM2 amplification in 41 cases of adipocytic tumors. Amplification was defined in both techniques as a MDM2/CEN12 ratio of 2 or greater. Eleven cases showed amplification with both FISH and CISH, and 26 cases showed no amplification with both methods. Two cases had discordant results between CISH and FISH, and two cases were not interpretable by CISH. CISH is advantageous for allowing pathologists to evaluate the histologic and molecular alterations occurring simultaneously in a specimen. Moreover, CISH is found to be more cost- and time-efficient when used with automation, and the signals do not quench over time. CISH technique is a reliable alternative to FISH in the evaluation of adipocytic tumors for MDM2 amplification. © 2014 Wiley Periodicals, Inc.

  14. Determination of HER2 amplification in primary breast cancer using dual-colour chromogenic in situ hybridization is comparable to fluorescence in situ hybridization: a European multicentre study involving 168 specimens

    Science.gov (United States)

    García-Caballero, Tomás; Grabau, Dorthe; Green, Andrew R; Gregory, John; Schad, Arno; Kohlwes, Elke; Ellis, Ian O; Watts, Sarah; Mollerup, Jens

    2010-01-01

    García-Caballero T, Grabau D, Green A R, Gregory J, Schad A, Kohlwes E, Ellis I O, Watts S & Mollerup J (2010) Histopathology56, 472–480 Determination of HER2 amplification in primary breast cancer using dual-colour chromogenic in situ hybridization is comparable to fluorescence in situ hybridization: a European multicentre study involving 168 specimens Aims: Fluorescence in situ hybridization (FISH) can be used to reveal several genomic imbalances relevant to proper cancer diagnosis and to the correct treatment regime. However, FISH requires expensive and advanced fluorescence microscopes in addition to expertise in fluorescence microscopy. To determine whether a newly developed dual-colour chromogenic in situ hybridization (CISH) method is a suitable alternative to FISH, we analysed the human epidermal growth factor receptor 2 gene (HER2) amplification level of 168 breast cancer specimens using dual-colour CISH and FISH and compared the results. Methods and results: We found 100% agreement between HER2 status determined by FISH and dual-colour CISH. Furthermore, we observed that the time used to score slides was significantly reduced by 28% in dual-colour CISH compared with the FISH protocol. Concordance between HER2 protein status and dual-colour CISH or FISH was equally good with an overall agreement of 96.8%. Correlation between the HER2/centromere 17 gene ratios obtained with dual-colour CISH and FISH was highly significant with an overall correlation coefficient (ρ) of 0.96. Conclusions: We conclude that dual-colour CISH and bright field microscopy are excellent alternatives to FISH when analysing the HER2 status of primary breast cancer. PMID:20459554

  15. Fluorescence In Situ Hybridization for MDM2 Amplification as a Routine Ancillary Diagnostic Tool for Suspected Well-Differentiated and Dedifferentiated Liposarcomas: Experience at a Tertiary Center

    Directory of Open Access Journals (Sweden)

    Khin Thway

    2015-01-01

    Full Text Available Background. The assessment of MDM2 gene amplification by fluorescence in situ hybridization (FISH has become a routine ancillary tool for diagnosing atypical lipomatous tumor (ALT/well-differentiated liposarcoma and dedifferentiated liposarcoma (WDL/DDL in specialist sarcoma units. We describe our experience of its utility at our tertiary institute. Methods. All routine histology samples in which MDM2 amplification was assessed with FISH over a 2-year period were included, and FISH results were correlated with clinical and histologic findings. Results. 365 samples from 347 patients had FISH for MDM2 gene amplification. 170 were positive (i.e., showed MDM2 gene amplification, 192 were negative, and 3 were technically unsatisfactory. There were 122 histologically benign cases showing a histology:FISH concordance rate of 92.6%, 142 WDL/DDL (concordance 96.5%, and 34 cases histologically equivocal for WDL (concordance 50%. Of 64 spindle cell/pleomorphic neoplasms (in which DDL was a differential diagnosis, 21.9% showed MDM2 amplification. Of the cases with discrepant histology and FISH, all but 3 had diagnoses amended following FISH results. For discrepancies of benign histology but positive FISH, lesions were on average larger, more frequently in “classical” (intra-abdominal or inguinal sites for WDL/DDL and more frequently core biopsies. Discrepancies of malignant histology but negative FISH were smaller, less frequently in “classical” sites but again more frequently core biopsies. Conclusions. FISH has a high correlation rate with histology for cases with firm histologic diagnoses of lipoma or WDL/DDL. It is a useful ancillary diagnostic tool in histologically equivocal cases, particularly in WDL lacking significant histologic atypia or DDL without corresponding WDL component, especially in larger tumors, those from intra-abdominal or inguinal sites or core biopsies. There is a significant group of well-differentiated adipocytic neoplasms

  16. USING THE US EXPERIENCE OF ONLINE AND HYBRID EDUCATION IN UKRAINIAN UNIVERSITIES

    Directory of Open Access Journals (Sweden)

    Ірина Задорожна

    2015-12-01

    Full Text Available The article investigates the best practices of the US universities on providing hybrid and online education that can be implemented at Ukrainian universities which experience lack of finances and a decrease in student enrollment. Basic factors of online and hybrid courses popularity are analysed (flexibility; accessibility, especially for students with special needs; saving time; convenience; motivation as students often feel less stressed in a virtual classroom than in a face-to-face environment. The main challenges of online and hybrid learning (faculty training and professional development for online education (in terms of pedagogy, communication and technology, supporting student learning in the online environment, and creating an efficient and interactive online learning community are defined in the article. Hybrid courses are regarded as such that promote some equivalence between digital and live communication.

  17. Comparison of Fluorescence In Situ Hybridization and Chromogenic In Situ Hybridization for Low and High Throughput HER2 Genetic Testing

    DEFF Research Database (Denmark)

    Poulsen, Tim S; Espersen, Maiken Lise Marcker; Kofoed, Vibeke

    2013-01-01

    cancer patients with HER2 immunohistochemistry (IHC) results scored as 0/1+, 2+, and 3+. HER2 genetic status was analysed using chromogenic in situ hybridization (CISH) and fluorescence in situ hybridization (FISH). Scoring results were documented through digital image analysis. The cancer region...

  18. Species integrity enhanced by a predation cost to hybrids in the wild

    DEFF Research Database (Denmark)

    Nilsson, P. A.; Hulthén, Kaj; Chapman, Ben B.

    2017-01-01

    Species integrity can be challenged, and even eroded, if closely related species can hybridize and produce fertile offspring of comparable fitness to that of parental species. The maintenance of newly diverged or closely related species therefore hinges on the establishment and effectiveness of pre...... barrier to gene flow in the wild. Cyprinid fishes commonly produce fertile, viable hybrid offspring and therefore make excellent study organisms to investigate ecological costs to hybrids. We electronically tagged two freshwater cyprinid fish species (roach Rutilus rutilus and bream Abramis brama...... to directly test for a predation cost to hybrids in the wild. Hybrid individuals were found significantly more susceptible to cormorant predation than individuals from either parental species. Such ecological selection against hybrids contributes to species integrity, and can enhance species diversification....

  19. Detection of dengue group viruses by fluorescence in situ hybridization

    Directory of Open Access Journals (Sweden)

    Raquin Vincent

    2012-10-01

    Full Text Available Abstract Background Dengue fever (DF and dengue hemorrhagic fever (DHF represent a global challenge in public health. It is estimated that 50 to 100 million infections occur each year causing approximately 20,000 deaths that are usually linked to severe cases like DHF and dengue shock syndrome. The causative agent of DF is dengue virus (genus Flavivirus that comprises four distinct serotypes (DENV-1 to DENV-4. Fluorescence in situ hybridization (FISH has been used successfully to detect pathogenic agents, but has not been implemented in detecting DENV. To improve our understanding of DENV infection and dissemination in host tissues, we designed specific probes to detect DENV in FISH assays. Methods Oligonucleotide probes were designed to hybridize with RNA from the broadest range of DENV isolates belonging to the four serotypes, but not to the closest Flavivirus genomes. Three probes that fit the criteria defined for FISH experiments were selected, targeting both coding and non-coding regions of the DENV genome. These probes were tested in FISH assays against the dengue vector Aedes albopictus (Diptera: Culicidae. The FISH experiments were led in vitro using the C6/36 cell line, and in vivo against dissected salivary glands, with epifluorescence and confocal microscopy. Results The three 60-nt oligonucleotides probes DENV-Probe A, B and C cover a broad range of DENV isolates from the four serotypes. When the three probes were used together, specific fluorescent signals were observed in C6/36 infected with each DENV serotypes. No signal was detected in either cells infected with close Flavivirus members West Nile virus or yellow fever virus. The same protocol was used on salivary glands of Ae. albopictus fed with a DENV-2 infectious blood-meal which showed positive signals in the lateral lobes of infected samples, with no significant signal in uninfected mosquitoes. Conclusion Based on the FISH technique, we propose a way to design and use

  20. Optimized Fast-FISH with a-satellite probes: acceleration by microwave activation

    Directory of Open Access Journals (Sweden)

    Durm M.

    1997-01-01

    Full Text Available It has been shown for several DNA probes that the recently introduced Fast-FISH (fluorescence in situ hybridization technique is well suited for quantitative microscopy. For highly repetitive DNA probes the hybridization (renaturation time and the number of subsequent washing steps were reduced considerably by omitting denaturing chemical agents (e.g., formamide. The appropriate hybridization temperature and time allow a clear discrimination between major and minor binding sites by quantitative fluorescence microscopy. The well-defined physical conditions for hybridization permit automatization of the procedure, e.g., by a programmable thermal cycler. Here, we present optimized conditions for a commercially available X-specific a-satellite probe. Highly fluorescent major binding sites were obtained for 74oC hybridization temperature and 60 min hybridization time. They were clearly discriminated from some low fluorescent minor binding sites on metaphase chromosomes as well as in interphase cell nuclei. On average, a total of 3.43 ± 1.59 binding sites were measured in metaphase spreads, and 2.69 ± 1.00 in interphase nuclei. Microwave activation for denaturation and hybridization was tested to accelerate the procedure. The slides with the target material and the hybridization buffer were placed in a standard microwave oven. After denaturation for 20 s at 900 W, hybridization was performed for 4 min at 90 W. The suitability of a microwave oven for Fast-FISH was confirmed by the application to a chromosome 1-specific a-satellite probe. In this case, denaturation was performed at 630 W for 60 s and hybridization at 90 W for 5 min. In all cases, the results were analyzed quantitatively and compared to the results obtained by Fast-FISH. The major binding sites were clearly discriminated by their brightness

  1. Expanding probe repertoire and improving reproducibility in human genomic hybridization

    Science.gov (United States)

    Dorman, Stephanie N.; Shirley, Ben C.; Knoll, Joan H. M.; Rogan, Peter K.

    2013-01-01

    Diagnostic DNA hybridization relies on probes composed of single copy (sc) genomic sequences. Sc sequences in probe design ensure high specificity and avoid cross-hybridization to other regions of the genome, which could lead to ambiguous results that are difficult to interpret. We examine how the distribution and composition of repetitive sequences in the genome affects sc probe performance. A divide and conquer algorithm was implemented to design sc probes. With this approach, sc probes can include divergent repetitive elements, which hybridize to unique genomic targets under higher stringency experimental conditions. Genome-wide custom probe sets were created for fluorescent in situ hybridization (FISH) and microarray genomic hybridization. The scFISH probes were developed for detection of copy number changes within small tumour suppressor genes and oncogenes. The microarrays demonstrated increased reproducibility by eliminating cross-hybridization to repetitive sequences adjacent to probe targets. The genome-wide microarrays exhibited lower median coefficients of variation (17.8%) for two HapMap family trios. The coefficients of variations of commercial probes within 300 nt of a repetitive element were 48.3% higher than the nearest custom probe. Furthermore, the custom microarray called a chromosome 15q11.2q13 deletion more consistently. This method for sc probe design increases probe coverage for FISH and lowers variability in genomic microarrays. PMID:23376933

  2. Genetic population structure of turbot ( Scophthalmus maximus L.) supports the presence of multiple hybrid zones for marine fishes in the transition zone between the Baltic Sea and the North Sea

    DEFF Research Database (Denmark)

    Eg Nielsen, Einar; Nielsen, P.H.; Meldrup, Dorte

    2004-01-01

    Sea, suggesting high gene flow among populations in these areas. In contrast, there was a sharp cline in genetic differentiation going from the low saline Baltic Sea to the high saline North Sea. The data were explained best by two divergent populations connected by a hybrid zone; however......, a mechanical mixing model could not be ruled out. A significant part of the genetic variance could be ascribed to variation among years within locality. Nevertheless, the population structure was relatively stable over time, suggesting that the observed pattern of genetic differentiation is biologically...... significant. This study suggests that hybrid zones are a common phenomenon for marine fishes in the transition area between the North Sea and the Baltic Sea and highlights the importance of using interspecific comparisons for inferring population structure in high gene flow species such as most marine fishes....

  3. Introgressive hybridization as a promoter of genome reshuffling in natural homoploid fish hybrids (Cyprinidae, Leuciscinae)

    Czech Academy of Sciences Publication Activity Database

    Pereira, C. S.; Aboim, M. A.; Ráb, Petr; Collares-Pereira, M. J.

    2014-01-01

    Roč. 112, č. 3 (2014), s. 343-350 ISSN 0018-067X Institutional support: RVO:67985904 Keywords : comparative genome hybridization * hybrid zones * introgression Subject RIV: EG - Zoology Impact factor: 3.805, year: 2014

  4. Australian experience of fish passage past instream structures

    International Nuclear Information System (INIS)

    Lewis, B.

    2008-01-01

    The growth in hydropower has resulted in the construction of various structures across rivers and streams, such as dams and weirs, which may impede essential fish movements and result in local extinctions of some fish species. When it is not practical to build instream structures that provide for fish passage, it may be appropriate to install some type of fishway. Site specific factors such as the fish species present, topography, flow characteristics and cost effectiveness will determine how best to provide for fish passage. The types of fishways suitable at small dams and weirs up to five metres high were described in this paper along with their benefits and effectiveness. The purpose was to provide simple and appropriate solutions that can improve the health of rivers considerably by managing the native aquatic habitat. The upstream passage past obstacles can be provided for through several types of fishways such as pool-type fishways, Denil fish passes, rock ramps, nature-like bypass channels, fish lifts or locks, collection and transportation facilities. In addition to environmental benefits, providing for fish passage can have long term social and economic benefits as well. 17 refs., 3 figs

  5. Identification of Cannabis sativa L. using the 1-kbTHCA synthase-fluorescence in situ hybridization probe.

    Science.gov (United States)

    Jeangkhwoa, Pattraporn; Bandhaya, Achirapa; Umpunjun, Puangpaka; Chuenboonngarm, Ngarmnij; Panvisavas, Nathinee

    2017-03-01

    This study reports a successful application of fluorescence in situ hybridization (FISH) technique in the identification of Cannabis sativa L. cells recovered from fresh and dried powdered plant materials. Two biotin-16-dUTP-labeled FISH probes were designed from the Cannabis-specific tetrahydrocannabinolic acid synthase (THCAS) gene and the ITS region of the 45S rRNA gene. Specificity of probe-target hybridization was tested against the target and 4 non-target plant species, i.e., Humulus lupulus, Mitragyna speciosa, Papaver sp., and Nicotiana tabacum. The 1-kb THCA synthase hybridization probe gave Cannabis-specific hybridization signals, unlike the 700-bp Cannabis-ITS hybridization probe. Probe-target hybridization was also confirmed against 20 individual Cannabis plant samples. The 1-kb THCA synthase and 700-bp Cannabis-ITS hybridization probes clearly showed 2 hybridization signals per cell with reproducibility. The 1-kb THCA synthase probe did not give any FISH signal when tested against H. lupulus, its closely related member of the Canabaceae family. It was also showed that 1-kb THCA synthase FISH probe can be applied to identify small amount of dried powdered Cannabis material with an addition of rehydration step prior to the experimental process. This study provided an alternative identification method for Cannabis trace. Copyright © 2016. Published by Elsevier B.V.

  6. Chromosomal imbalances detected in primary bone tumors by comparative genomic hybridization and interphase fluorescence in situ hybridization

    Directory of Open Access Journals (Sweden)

    Marcelo Razera Baruffi

    2003-01-01

    Full Text Available We applied a combination of comparative genomic hybridization (CGH and fluorescence in situ hybridization (FISH, to characterize the genetic aberrations in three osteosarcomas (OS and one Ewing's sarcoma. CGH identified recurrent chromosomal losses at 10p14-pter and gains at 8q22.3-24.1 in OS. Interphase FISH allowed to confirm 8q gain in two cases. A high amplification level of 11q12-qter was detected in one OS. The Ewing's sarcoma showed gain at 1p32-36.1 as the sole chromosome alteration. These studies demonstrate the value of molecular cytogenetic methods in the characterization of recurrent genomic alterations in bone tumor tissue.

  7. Chromogenic in situ hybridization for Her-2/neu-oncogene in breast cancer: comparison of a new dual-colour chromogenic in situ hybridization with immunohistochemistry and fluorescence in situ hybridization.

    Science.gov (United States)

    Mayr, Doris; Heim, Sibylle; Weyrauch, Kerstin; Zeindl-Eberhart, Evelyn; Kunz, Anne; Engel, Jutta; Kirchner, Thomas

    2009-12-01

    Her-2/neu testing is used as a marker for Herceptin therapy. The aim was to investigate new dual-colour chromogenic in situ hybridization (CISH), in a large number of breast carcinomas (n = 205) with DNA-specific dual-colour probes (ZytoVision, Bremerhaven, Germany) and to compare the results with immunohistochemistry (n = 205) and fluorescence in situ hybridization (FISH) (n = 129). Paraffin-embedded tissue of 205 patients was used. After immunohistochemistry with a focus on immunohistochemically uncertain cases, Her-2/neu amplification using dual-colour CISH (ZytoVision) was analysed. Validation by FISH was performed. The results were: immunohistochemistry, 27.8% with strong expression, 53.7% with uncertain overexpression and 18.5% with no expression; FISH, 25.6% amplified and 74.4% negative; CISH, 35.6% amplified, 62.9% negative and 1.5% not evaluable. Comparison of immunohistochemistry with CISH: CISH negative in 100% with immunohistochemistry 0/1+, amplified in 82.5% with immunohistochemistry 3+; 5.9% contradictory results: 4.4% immunohistochemistry 3+ and negative by CISH, 1.5% negative in immunohistochemistry but amplified by CISH; FISH (129 cases), 8.5% contradictory results to immunohistochemistry, 6.2% immunohistochemistry 3+ and negative by FISH, 2.3% negative by immunohistochemistry and amplified by FISH; comparison of CISH and FISH, 94.6% same results, 3.9% different ones, 1.6% CISH not analysable. CISH, using dual-colour probes (ZytoVision) is as good as FISH for Her-2/neu analysis. The few discrepant results are likely to be caused by polysomy or tumour heterogeneity.

  8. Fluorescence in situ hybridization with reference to biodosimetry: a review

    International Nuclear Information System (INIS)

    Venkatachalam, P.; Paul, S.F.D.; Jeevanram, R.K.

    1996-01-01

    Many advances have taken place in the field of radiation biodosimetry in the recent past. Measurement of dicentric chromosome aberrations, was first developed and followed by micronuclei scoring. These, however, are unstable type aberrations and the cells carrying such aberrations are eliminated from the body in few years. They are therefore of use primarily in case of accidental exposures. The challenge is to measure the cumulative radiation exposure resulting from normal operations by measuring stable chromosome aberrations. Banding technique can measure stable chromosome aberration but require long time to analyse the banding pattern to study translocations. On the other hand fluorescence in situ hybridization (FISH) technique is sensitive, fast and easy to identify the translocations as the chromosomes involved in translocation are painted with different colours. This review brings out the requirements of various materials, their preparations, method of detection of fluorescence etc. for carrying out FISH. The experience of various laboratories using FISH in the monitoring of radiation absorbed dose is discussed. (author)

  9. Annual trends in catchability and fish stock assessments

    Directory of Open Access Journals (Sweden)

    Marchal Paul

    2003-04-01

    Full Text Available A key assumption of many fish stock assessment models is that catchability is constant over time. We assume here that trends in catchability may occur through fishing power creeping. The tuning fleets, which are prone to fishing power development, may be identified using the Hybrid method. A range of catchability trends, including values derived from the Hybrid method, is then implemented to standardise the fishing effort of some tuning fleets used in the stock assessments performed by XSA (eXtended Survivors Analysis. Stocks being assessed are the North Sea cod, saithe, plaice and sole. The performances of the new and traditional XSA assessments are compared using criteria based on the precision of catchability estimates, stationarity of Log-catchability residuals and retrospective patterns relative to fishing mortality, spawning stock biomass and recruitment estimates. The performances of the North Sea cod, plaice and sole assessments could be enhanced by accounting for an overall annual increase in the catchability of some of the tuning fleets. No significant trends could be detected in the catchability of the tuning fleets relative to the assessment of the North Sea saithe. By contrast with the traditional assessment, the spawning biomass of cod is expected not to have increased between 1997 and 1998, while the fishing mortality of sole is expected to have increased over the same period.

  10. Asexual Reproduction Does Not Apparently Increase the Rate of Chromosomal Evolution: Karyotype Stability in Diploid and Triploid Clonal Hybrid Fish (Cobitis, Cypriniformes, Teleostei).

    Science.gov (United States)

    Majtánová, Zuzana; Choleva, Lukáš; Symonová, Radka; Ráb, Petr; Kotusz, Jan; Pekárik, Ladislav; Janko, Karel

    2016-01-01

    Interspecific hybridization, polyploidization and transitions from sexuality to asexuality considerably affect organismal genomes. Especially the last mentioned process has been assumed to play a significant role in the initiation of chromosomal rearrangements, causing increased rates of karyotype evolution. We used cytogenetic analysis and molecular dating of cladogenetic events to compare the rate of changes of chromosome morphology and karyotype in asexually and sexually reproducing counterparts in European spined loach fish (Cobitis). We studied metaphases of three sexually reproducing species and their diploid and polyploid hybrid clones of different age of origin. The material includes artificial F1 hybrid strains, representatives of lineage originated in Holocene epoch, and also individuals of an oldest known age to date (roughly 0.37 MYA). Thereafter we applied GISH technique as a marker to differentiate parental chromosomal sets in hybrids. Although the sexual species accumulated remarkable chromosomal rearrangements after their speciation, we observed no differences in chromosome numbers and/or morphology among karyotypes of asexual hybrids. These hybrids possess chromosome sets originating from respective parental species with no cytogenetically detectable recombinations, suggesting their integrity even in a long term. The switch to asexual reproduction thus did not provoke any significant acceleration of the rate of chromosomal evolution in Cobitis. Asexual animals described in other case studies reproduce ameiotically, while Cobitis hybrids described here produce eggs likely through modified meiosis. Therefore, our findings indicate that the effect of asexuality on the rate of chromosomal change may be context-dependent rather than universal and related to particular type of asexual reproduction.

  11. Asexual Reproduction Does Not Apparently Increase the Rate of Chromosomal Evolution: Karyotype Stability in Diploid and Triploid Clonal Hybrid Fish (Cobitis, Cypriniformes, Teleostei.

    Directory of Open Access Journals (Sweden)

    Zuzana Majtánová

    Full Text Available Interspecific hybridization, polyploidization and transitions from sexuality to asexuality considerably affect organismal genomes. Especially the last mentioned process has been assumed to play a significant role in the initiation of chromosomal rearrangements, causing increased rates of karyotype evolution. We used cytogenetic analysis and molecular dating of cladogenetic events to compare the rate of changes of chromosome morphology and karyotype in asexually and sexually reproducing counterparts in European spined loach fish (Cobitis. We studied metaphases of three sexually reproducing species and their diploid and polyploid hybrid clones of different age of origin. The material includes artificial F1 hybrid strains, representatives of lineage originated in Holocene epoch, and also individuals of an oldest known age to date (roughly 0.37 MYA. Thereafter we applied GISH technique as a marker to differentiate parental chromosomal sets in hybrids. Although the sexual species accumulated remarkable chromosomal rearrangements after their speciation, we observed no differences in chromosome numbers and/or morphology among karyotypes of asexual hybrids. These hybrids possess chromosome sets originating from respective parental species with no cytogenetically detectable recombinations, suggesting their integrity even in a long term. The switch to asexual reproduction thus did not provoke any significant acceleration of the rate of chromosomal evolution in Cobitis. Asexual animals described in other case studies reproduce ameiotically, while Cobitis hybrids described here produce eggs likely through modified meiosis. Therefore, our findings indicate that the effect of asexuality on the rate of chromosomal change may be context-dependent rather than universal and related to particular type of asexual reproduction.

  12. Apparent digestibility of Asian carp and common carp-derived fish meals in feeds for hybrid striped bass Morone saxatilis female x M. chrysops male and rainbow trout Oncorhynchus mykiss

    Science.gov (United States)

    Apparent digestibility coefficients (ADCs) of nutrients (crude protein, amino acids, crude lipid, fatty acids, and minerals) were determined for fish meals derived from menhaden, Asian carp (combination of silver and bighead carps), and common carp in feeds for hybrid striped bass and rainbow trout....

  13. Hybridization experiments indicate incomplete reproductive isolating mechanism between Fasciola hepatica and Fasciola gigantica.

    Science.gov (United States)

    Itagaki, T; Ichinomiya, M; Fukuda, K; Fusyuku, S; Carmona, C

    2011-09-01

    Experiments on hybridization between Fasciola hepatica and Fasciola gigantica were carried out to clarify whether a reproductive isolating mechanism appears between the two Fasciola species. Molecular evidence for hybridization was based on the DNA sequence of the internal transcribed spacer 1 (ITS1) region in nuclear ribosomal DNA, which differs between the species. The results suggested that there were not pre-mating but post-mating isolating mechanisms between the two species. However, viable adults of the hybrids F1 and F2 were produced from both parental F. hepatica and F. gigantica. The hybrids inherited phenotypic characteristics such as ratio of body length and width and infectivity to rats from parental Fasciola hepatica and F. gigantica. These findings suggest that reproductive isolation is incomplete between Fasciola hepatica and F. gigantica. Adults of the hybrids F1 and F2 were completely different in mode of reproduction from aspermic Fasciola forms that occur in Asia and seem to be offspring originated from hybridization between F. hepatica and F. gigantica and to reproduce parthenogenetically.

  14. Students' Positive and Negative Experiences in Hybrid and Online Classes

    Science.gov (United States)

    El Mansour, Bassou; Mupinga, Davison M.

    2007-01-01

    As higher education institutions struggle to meet the growing demand for education from non-traditional students, many are turning to hybrid and online courses. These courses, free up classroom space, allow faculty to reach a wider audience using technology; and are therefore cost effective. But, what learning experiences do these courses provide…

  15. Validation of microsatellite multiplexes for parentage analysis and species discrimination in two hybridizing species of coral reef fish (Plectropomus spp., Serranidae)

    KAUST Repository

    Harrison, H.B.

    2014-04-24

    Microsatellites are often considered ideal markers to investigate ecological processes in animal populations. They are regularly used as genetic barcodes to identify species, individuals, and infer familial relationships. However, such applications are highly sensitive the number and diversity of microsatellite markers, which are also prone to error. Here, we propose a novel framework to assess the suitability of microsatellite datasets for parentage analysis and species discrimination in two closely related species of coral reef fish, Plectropomus leopardus and P. maculatus (Serranidae). Coral trout are important fisheries species throughout the Indo-Pacific region and have been shown to hybridize in parts of the Great Barrier Reef, Australia. We first describe the development of 25 microsatellite loci and their integration to three multiplex PCRs that co-amplify in both species. Using simulations, we demonstrate that the complete suite of markers provides appropriate power to discriminate between species, detect hybrid individuals, and resolve parent-offspring relationships in natural populations, with over 99.6% accuracy in parent-offspring assignments. The markers were also tested on seven additional species within the Plectropomus genus with polymorphism in 28-96% of loci. The multiplex PCRs developed here provide a reliable and cost-effective strategy to investigate evolutionary and ecological dynamics and will be broadly applicable in studies of wild populations and aquaculture brood stocks for these closely related fish species. 2014 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.

  16. Validation of microsatellite multiplexes for parentage analysis and species discrimination in two hybridizing species of coral reef fish (Plectropomus spp., Serranidae)

    KAUST Repository

    Harrison, H.B.; Feldheim, K.A.; Jones, G.P.; Ma, K.; Mansour, H.; Perumal, S.; Williamson, D.H.; Berumen, Michael L.

    2014-01-01

    Microsatellites are often considered ideal markers to investigate ecological processes in animal populations. They are regularly used as genetic barcodes to identify species, individuals, and infer familial relationships. However, such applications are highly sensitive the number and diversity of microsatellite markers, which are also prone to error. Here, we propose a novel framework to assess the suitability of microsatellite datasets for parentage analysis and species discrimination in two closely related species of coral reef fish, Plectropomus leopardus and P. maculatus (Serranidae). Coral trout are important fisheries species throughout the Indo-Pacific region and have been shown to hybridize in parts of the Great Barrier Reef, Australia. We first describe the development of 25 microsatellite loci and their integration to three multiplex PCRs that co-amplify in both species. Using simulations, we demonstrate that the complete suite of markers provides appropriate power to discriminate between species, detect hybrid individuals, and resolve parent-offspring relationships in natural populations, with over 99.6% accuracy in parent-offspring assignments. The markers were also tested on seven additional species within the Plectropomus genus with polymorphism in 28-96% of loci. The multiplex PCRs developed here provide a reliable and cost-effective strategy to investigate evolutionary and ecological dynamics and will be broadly applicable in studies of wild populations and aquaculture brood stocks for these closely related fish species. 2014 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.

  17. HER2 Gene Amplification Testing by Fluorescent In Situ Hybridization (FISH): Comparison of the ASCO-College of American Pathologists Guidelines With FISH Scores Used for Enrollment in Breast Cancer International Research Group Clinical Trials.

    Science.gov (United States)

    Press, Michael F; Sauter, Guido; Buyse, Marc; Fourmanoir, Hélène; Quinaux, Emmanuel; Tsao-Wei, Denice D; Eiermann, Wolfgang; Robert, Nicholas; Pienkowski, Tadeusz; Crown, John; Martin, Miguel; Valero, Vicente; Mackey, John R; Bee, Valerie; Ma, Yanling; Villalobos, Ivonne; Campeau, Anaamika; Mirlacher, Martina; Lindsay, Mary-Ann; Slamon, Dennis J

    2016-10-10

    Purpose ASCO and the College of American Pathologists (ASCO-CAP) recently recommended further changes to the evaluation of human epidermal growth factor receptor 2 gene (HER2) amplification by fluorescent in situ hybridization (FISH). We retrospectively assessed the impact of these new guidelines by using annotated Breast Cancer International Research Group (BCIRG) -005, BCIRG-006, and BCIRG-007 clinical trials data for which we have detailed outcomes. Patients and Methods The HER2 FISH status of BCIRG-005/006/007 patients with breast cancers was re-evaluated according to current ASCO-CAP guidelines, which designates five different groups according to HER2 FISH ratio and average HER2 gene copy number per tumor cell: group 1 (in situ hybridization [ISH]-positive): HER2-to-chromosome 17 centromere ratio ≥ 2.0, average HER2 copies ≥ 4.0; group 2 (ISH-positive): ratio ≥ 2.0, copies < 4.0; group 3 (ISH-positive): ratio < 2.0, copies ≥ 6.0; group 4 (ISH-equivocal): ratio < 2.0, copies ≥ 4.0 and < 6.0; and group 5 (ISH-negative): ratio < 2.0, copies < 4.0. We assessed correlations with HER2 protein, clinical outcomes by disease-free survival (DFS) and overall survival (OS) and benefit from trastuzumab therapy (hazard ratio [HR]). Results Among 10,468 patients with breast cancers who were successfully screened for trial entry, 40.8% were in ASCO-CAP ISH group 1, 0.7% in group 2; 0.5% in group 3, 4.1% in group 4, and 53.9% in group 5. Distributions were similar in screened compared with accrued subpopulations. Among accrued patients, FISH group 1 breast cancers were strongly correlated with immunohistochemistry 3+ status (P < .0001), whereas groups 2, 3, 4, and 5 were not; however, groups 2, 4 and, 5 were strongly correlated with immunohistochemistry 0/1+ status (all P < .0001), whereas group 3 was not. Among patients accrued to BCIRG-005, group 4 was not associated with significantly worse DFS or OS compared with group 5. Among patients accrued to BCIRG-006, only

  18. Dual-colour chromogenic in-situ hybridization is a potential alternative to fluorescence in-situ hybridization in HER2 testing.

    Science.gov (United States)

    Hwang, Cheng-Cheng; Pintye, Mariann; Chang, Liang-Che; Chen, Huang-Yang; Yeh, Kun-Yan; Chein, Hui-Ping; Lee, Nin; Chen, Jim-Ray

    2011-11-01

    Dual-colour chromogenic in-situ hybridization (dc-CISH) is an emerging methodology for characterizing genomic alterations. This study was aimed at evaluating the performance of a dc-CISH kit (ZytoVision) in determining human epidermal growth factor receptor 2 (HER2) status in breast cancer. Two hundred and twenty-eight invasive breast carcinomas arranged in tissue microarrays were analysed in parallel with dc-CISH, fluorescence in-situ hybridization (FISH), and immunohistochemistry. Of 227 tumours with available FISH and dc-CISH results, HER2 amplification and non-amplification were detected in 49 (21.6%) and 178 (78.4%) tumours, respectively, by both assays. The concordance between dc-CISH and FISH results showed 100% agreement (κ-coefficient=1.00). Immunohistochemically, 162 (71%), 25 (11.0%) and 41 (18%) tumours were scored 0/1+, 2+, and 3+, respectively. The corresponding results with both FISH and dc-CISH demonstrated HER2 amplification in two (3.2%), nine (36%) and 38 (93%) tumours, respectively. Complete consensus among these three methods was observed in 197 cases, representing 98% of all 3+ and 0/1+ tumours (κ-coefficient=0.92). Confirmatory testing of 25 2+ tumours showed complete consensus between FISH and dc-CISH. dc-CISH is a promising alternative to FISH in HER2 testing, and the single-institute incidence of HER2 amplification in breast cancer in Taiwan is 21.2%. © 2011 Blackwell Publishing Limited.

  19. Perulernaea gamitanae (Copepoda: Lernaeidae parasitizing tambaqui (Colossoma macropomum (Characidae and the hybrids tambacu and tambatinga, cultured in northern Brazil

    Directory of Open Access Journals (Sweden)

    M. Tavares-Dias

    2011-08-01

    Full Text Available The infestation rate in Colossoma macropomum, hybrid tambacu (C. macropomum x Piaractus mesopotamicus and hybrid tambatinga (C. macropomum x Piaractus brachypomum with Perulernaea gamitanae Thatcher and Paredes, 1985 from two fish farms in Amapá State, Brazil was studied. Lernaeid parasites (n=2887 were collected mainly on the tongue and the mouth cavity and also on cartilage of gill arches and filaments. Inflammation and fibrous nodules were observed on the attachment sites of the parasites. The infestation rate varied according to the fish farm and host. The prevalence of P. gamitanae was of 100% in hosts from one fish farm and was lower in the other fish farm. Higher intensity of P. gamitanae occurred in hybrids tambacu and tambatinga, but despite the high prevalence its intensity was moderate. This is the first report on epidemiology of P. gamitanae in cultured fishes from Brazilian Amazonia, and the occurrence of this crustacean parasite in two new hosts, the hybrids tambacu and tambatinga.

  20. Combining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions

    Directory of Open Access Journals (Sweden)

    Yusuf Mohammed

    2011-12-01

    Full Text Available Abstract Background Physical mapping of transgenic insertions by Fluorescence in situ Hybridization (FISH is a reliable and cost-effective technique. Chromosomal assignment is commonly achieved either by concurrent G-banding or by a multi-color FISH approach consisting of iteratively co-hybridizing the transgenic sequence of interest with one or more chromosome-specific probes at a time, until the location of the transgenic insertion is identified. Results Here we report a technical development for fast chromosomal assignment of transgenic insertions at the single cell level in mouse and rat models. This comprises a simplified 'single denaturation mixed hybridization' procedure that combines multi-color karyotyping by Multiplex FISH (M-FISH, for simultaneous and unambiguous identification of all chromosomes at once, and the use of a Quantum Dot (QD conjugate for the transgene detection. Conclusions Although the exploitation of the unique optical properties of QD nanocrystals, such as photo-stability and brightness, to improve FISH performance generally has been previously investigated, to our knowledge this is the first report of a purpose-designed molecular cytogenetic protocol in which the combined use of QDs and standard organic fluorophores is specifically tailored to assist gene transfer technology.

  1. Determination of 24 personal care products in fish bile using hybrid solvent precipitation and dispersive solid phase extraction cleanup with ultrahigh performance liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometry.

    Science.gov (United States)

    Yao, Li; Lv, Yin-Zhi; Zhang, Li-Juan; Liu, Wang-Rong; Zhao, Jian-Liang; Liu, You-Sheng; Zhang, Qian-Qian; Ying, Guang-Guo

    2018-05-25

    Personal care products (PCPs) are ubiquitous in aquatic environments owing to the continuous discharge of domestic wastewater from highly urbanized regions. These PCPs can be adsorbed by fish and thereafter usually enter the bile of the fish through biliary excretion. In this study, a sensitive method based on a combination of hybrid solvent precipitation and dispersive solid phase extraction (d-SPE) purification was developed to simultaneously extract and detect 24 PCPs, namely, 16 biocides, 4 synthetic musks, and 4 benzotriazoles, from fish bile. Hybrid precipitation on solid phase extraction (SPE) tubes was applied to remove phospholipids and proteins, and a d-SPE procedure was used for further purification. The extraction solvents for the hybrid precipitation/SPE tubes and d-SPE materials were optimized. The method performance for bile samples both with and without enzyme hydrolysis using β-glucuronidase/aryl-sulfatase were validated. The 24 PCPs in fish bile were spiked with standard concentrations of 10 ng/mL, 20 ng/mL, 100 ng/mL, and 200 ng/mL to evaluate recoveries, which ranged from 70 to 120% for 16, 16, 22, and 21 analytes with hydrolysis, respectively, and 70-120% for 14, 15, 23, and 23 analytes without hydrolysis, respectively. The quantification limits for target PCPs were in the range 0.26-7.38 ng/mL [excluding musk xylene (MX) and musk ketone (MK)] and 0.20-9.48 ng/mL (excluding MX and MK) for bile samples with and without enzyme hydrolysis, respectively. After enzyme hydrolysis, 12 PCPs were detected in bile from fish collected from the Yangtze River, with a maximum detected concentration of 460 ng/mL, for triclosan (TCS). The hydrolysis reaction indicated that high percentages of glucuronide and sulfate metabolites for some PCPs, i.e. four parabens and TCS, existed in the bile. Copyright © 2018 Elsevier B.V. All rights reserved.

  2. Lower hybrid current drive experiments with graphite limiters in the HT-7 superconducting tokamak

    International Nuclear Information System (INIS)

    Liu, J.; Gao, X.; Hu, L.Q.; Asif, M.; Chen, Z.Y.; Ding, B.J.; Zhou, Q.; Liu, H.Q.; Jie, Y.X.; Kong, W.; Lin, S.Y.; Ding, Y.H.; Gao, L.; Xu, Q.

    2006-01-01

    Recent progress of lower hybrid (LH) experiments with new graphite limiters configuration in the HT-7 tokamak is presented. The lower hybrid current drive (LHCD) efficiency can be determined by fitting based on experimental data. Improved particle confinement was observed via LHCD (P LHW >300 kW) characterized by the particle confinement time τ p increased about 1.56 times. It is found that runaways are suppressed during loop voltage is decreasing at the flat-top phase of LH discharges. The main limitations of pulse length are presented in long-pulse experiments with new limiter configuration

  3. RNA detection in situ with FISH-STICs.

    Science.gov (United States)

    Sinnamon, John R; Czaplinski, Kevin

    2014-02-01

    The ability to detect RNA molecules in situ has long had important applications for molecular biological studies. Enzyme or dye-labeled antisense in vitro runoff transcripts and synthetic oligodeoxynucleotides (ODN) both have a proven track record of success, but each of these also has scientific and practical drawbacks and limitations to its use. We devised a means to use commercially synthesized oligonucleotides as RNA-FISH probes without further modification and show that such probes work well for detection of RNA in cultured cells. This approach can bind a high concentration of fluorescent ODN to a short stretch of an RNA using commercial DNA synthesis outlets available to any laboratory. We call this approach for creating in situ hybridization probes Fluorescence In Situ Hybridization with Sequential Tethered and Intertwined ODN Complexes (FISH-STICs). We demonstrate that one FISH-STIC probe can detect mRNA molecules in culture, and that probe detection can be improved by the addition of multiple probes that can be easily adapted for robust mRNA quantification. Using FISH-STICs, we demonstrate a nonoverlapping distribution for β-actin and γ-actin mRNA in cultured fibroblasts, and the detection of neuron-specific transcripts within cultured primary hippocampal neurons.

  4. Fluorescence In situ Hybridization: Cell-Based Genetic Diagnostic and Research Applications.

    Science.gov (United States)

    Cui, Chenghua; Shu, Wei; Li, Peining

    2016-01-01

    Fluorescence in situ hybridization (FISH) is a macromolecule recognition technology based on the complementary nature of DNA or DNA/RNA double strands. Selected DNA strands incorporated with fluorophore-coupled nucleotides can be used as probes to hybridize onto the complementary sequences in tested cells and tissues and then visualized through a fluorescence microscope or an imaging system. This technology was initially developed as a physical mapping tool to delineate genes within chromosomes. Its high analytical resolution to a single gene level and high sensitivity and specificity enabled an immediate application for genetic diagnosis of constitutional common aneuploidies, microdeletion/microduplication syndromes, and subtelomeric rearrangements. FISH tests using panels of gene-specific probes for somatic recurrent losses, gains, and translocations have been routinely applied for hematologic and solid tumors and are one of the fastest-growing areas in cancer diagnosis. FISH has also been used to detect infectious microbias and parasites like malaria in human blood cells. Recent advances in FISH technology involve various methods for improving probe labeling efficiency and the use of super resolution imaging systems for direct visualization of intra-nuclear chromosomal organization and profiling of RNA transcription in single cells. Cas9-mediated FISH (CASFISH) allowed in situ labeling of repetitive sequences and single-copy sequences without the disruption of nuclear genomic organization in fixed or living cells. Using oligopaint-FISH and super-resolution imaging enabled in situ visualization of chromosome haplotypes from differentially specified single-nucleotide polymorphism loci. Single molecule RNA FISH (smRNA-FISH) using combinatorial labeling or sequential barcoding by multiple round of hybridization were applied to measure mRNA expression of multiple genes within single cells. Research applications of these single molecule single cells DNA and RNA FISH

  5. Fluorescence In situ Hybridization: Cell-Based Genetic Diagnostic and Research Applications

    Directory of Open Access Journals (Sweden)

    Chenghua Cui

    2016-09-01

    Full Text Available Fluorescence in situ hybridization (FISH is a macromolecule recognition technology based on the complementary nature of DNA or DNA/RNA double strands. Selected DNA strands incorporated with fluorophore-coupled nucleotides can be used as probes to hybridize onto the complementary sequences in tested cells and tissues and then visualized through a fluorescence microscope or an imaging system. This technology was initially developed as a physical mapping tool to delineate genes within chromosomes. Its high analytical resolution to a single gene level and high sensitivity and specificity enabled an immediate application for genetic diagnosis of constitutional common aneuploidies, microdeletion/microduplication syndromes and subtelomeric rearrangements. FISH tests using panels of gene-specific probes for somatic recurrent losses, gains and translocations have been routinely applied for hematologic and solid tumors and are one of the fastest-growing areas in cancer diagnosis. FISH has also been used to detect infectious microbials and parasites like malaria in human blood cells. Recent advances in FISH technology involve various methods for improving probe labeling efficiency and the use of super resolution imaging systems for direct visualization of intra-nuclear chromosomal organization and profiling of RNA transcription in single cells. Cas9-mediated FISH (CASFISH allowed in situ labeling of repetitive sequences and single-copy sequences without the disruption of nuclear genomic organization in fixed or living cells. Using oligopaint-FISH and super-resolution imaging enabled in situ visualization of chromosome haplotypes from differentially specified single-nucleotide polymorphism loci. Single molecule RNA FISH (smRNA-FISH using combinatorial labeling or sequential barcoding by multiple round of hybridization were applied to measure mRNA expression of multiple genes within single cells. Research applications of these single molecule single cells

  6. Comparative analysis of the aquaculture potential of hybrid Tilapia ...

    African Journals Online (AJOL)

    , on the growth rate, feeding efficiency and mortality rates of hybrid tilapia — Tilapia zillii (male) x T. guineensis (female) — was evaluated for 233 days. Fish of average weight 12.59g were stocked at a density of 20 fish m–³ and were fed a 30% ...

  7. In Situ Hybridization Pada Kanker Payudara

    OpenAIRE

    Diah Witari, Ni Putu

    2014-01-01

    Kesulitan yang dijumpai pada penanganan kanker payudara adalah terjadinya kekambuhan atau relaps. Deteksi status HER2 pada pasien merupakan salah satu upaya untuk mendeteksi terjadinya relaps dan juga untuk menentukan jenis terapi yang ada diberikan. Ekspresi protein HER2 dapat dideteksi dengan immunohistochemistry (IHC), sedangkan mutasi gen HER2 dapat dideteksi dengan teknik in situ hybridization baik berupa fluorescence in situ hybridization (FISH) ataupun chromogenic in situ hy...

  8. Immunohistochemistry and alternative FISH testing in breast cancer with HER2 equivocal amplification.

    Science.gov (United States)

    Agersborg, Sally; Mixon, Christopher; Nguyen, Thanh; Aithal, Sramila; Sudarsanam, Sucha; Blocker, Forrest; Weiss, Lawrence; Gasparini, Robert; Jiang, Shiping; Chen, Wayne; Hess, Gregory; Albitar, Maher

    2018-03-22

    While HER2 testing is well established in directing appropriate treatment for breast cancer, a small percentage of cases show equivocal results by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Alternative probes may be used in equivocal cases. We present a single community-based institution's experience in further evaluating these cases. Between 2014 and 2016, 4255 samples were submitted for HER2 amplification testing by alternative probes, TP53, RAI1, and RARA. Of the patients tested by FISH, 505/3908 (12.9%) also had IHC data. Most (73.9%) FISH equivocal cases remained equivocal after IHC testing. However, 50.5% of equivocal cases were classified as HER2 amplified by alternative probes. Most cases were positive by more than one probe: 78% of positive cases by RAI1 and 73.9% by TP53. There was a significant difference between IHC and FISH alternative testing (p alternative FISH testing. Available data showed that 41% of patients were treated with palbociclib and were positive by alternative FISH. The prevalence of double HER2 equivocal cases and the discrepancy between IHC and alternative FISH testing suggest that FISH alternative testing using both RAI1 and TP53 probes is necessary for conclusive classification. Because almost half of FISH equivocal cases converted to HER2 amplified upon alternative testing, clinical studies to determine the benefit of anti-HER2 therapy in these patients are urgently needed.

  9. FISH and PNA FISH for the diagnosis of Q fever endocarditis and vascular infections.

    Science.gov (United States)

    Prudent, Elsa; Lepidi, Hubert; Angelakis, Emmanouil; Raoult, Didier

    2018-06-13

    Purpose. Endocarditis and vascular infections are common manifestations of persistent localized infection due to Coxiella burnetii and recently, fluorescence in situ hybridization (FISH) was proposed as an alternative tool for their diagnosis. In this study, we evaluated the efficiency of FISH in a series of valve and vascular samples infected by C. burnetii. Methods. We tested 23 C. burnetii -positive valves and thrombus samples obtained from patients with Q fever endocarditis. Seven aneurysms and thrombus specimens were retrieved from patients with Q fever vascular infection. Samples were analyzed by culture, immunochemistry and FISH with oligonucleotide and PNA probes targeting C. burnetii -specific 16S ribosomal RNA sequences. Results. Immunohistochemical analysis was positive for five (17%) samples with significantly more copies of C. burnetii DNA than the negative ones ( p= 0.02). FISH was positive for 13 (43%) samples and presented 43% and 40% sensitivity compared to qPCR and culture, respectively. PNA FISH detected C. burnetii in 18 (60%) samples and presented 60% and 55% sensitivity compared to qPCR and culture, respectively. Immunohistochemistry had 38% and 28% sensitivity compared to FISH and PNA FISH, respectively. Samples found positive by both immunohistochemistry and PNA FISH contained significantly more copies of C. burnetii DNA than the negative ones ( p= 0.03). Finally, PNA FISH was more sensitive than FISH (60% versus 43%) for the detection of C. burnetii Conclusion. We provide evidence that PNA FISH and FISH are important assays for the diagnosis of C. burnetii endocarditis and vascular infections. Copyright © 2018 American Society for Microbiology.

  10. A populational survey of 45S rDNA polymorphism in the Jefferson salamander Ambystoma jeffersonianum revealed by fluorescence in situ hybridization (FISH

    Directory of Open Access Journals (Sweden)

    Jinzhong FU

    2009-04-01

    Full Text Available The chromosomal localization of 45S ribosomal RNA genes in Ambystoma jeffersonianum was determined by fluorescence in situ hybridization with 18S rDNA fragment as a probe (FISH-rDNA. Our results revealed the presence of rDNA polymorphism among A.jeffersonianum populations in terms of number, location and FISH signal intensity on the chromosomes. Nine rDNA cytotypes were found in ten geographically isolated populations and most of them contained derivative rDNA sites. Our preliminary study provides strong indication of karyotypic diversification of A.jeffersonianum that is demonstrated by intraspecific variation of 45S rDNA cytotypes. rDNA cytotype polymorphism has been described in many other caudate amphibians. We predict that habitat isolation, low dispersal ability and decline of effective population size could facilitate the fixation and accumulation of variable rDNA cytotypes during their chromosome evolution.

  11. Lower hybrid heating experiment in JFT-2 tokamak

    International Nuclear Information System (INIS)

    Uchara, K.; Nagashima, T.

    1982-01-01

    Lower hybrid heating experiments in JFT-2 are reviewed. Good maintenance and controlling of the coupling structure are very important in the injection of RF power before heating experiments. Accessibility of waves and the existence of the mode conversion region are necessary for ion heating in the main plasma. Parametric instabilities which may bring undesirable power deposition are suppressed by enough electron heating in the boundary region. Optimizing the Nsub(z) spectrum and the improvement of the plasma confinement may lead the electron heating in the high density region. Current generation by use of quasi-linear Landau damping is confirmed and is suggested to bring the improvement of plasma confinement. High power and long pulse klystrons may be expected to open a frontier toward a stational reactor plasma in tokamaks. (author)

  12. Review of lower hybrid results in the Wega and Petula experiments

    International Nuclear Information System (INIS)

    Briffod, G.

    1980-04-01

    Along these lines, a large part of the experimental program of the Grenoble laboratory has been devoted for several years to the study of heating at the lower hybrid frequency, first on the WEGA experiment, (a cooperative experiment with IPP GARCHING) and, more recently, also on the PETULA device. In particular, at HF heating power of the order of the ohmic heating power, significant ion heating (increase of ion temperature by 100%) and electron heating (electron temperature increase by 60%) have been observed on the WEGA device and published in 1976 and 1978. Nevertheless, despite the increasing number of experimental results, detailed investigations of the power balance during the HF heating pulse have remained scarce. Consequently, several important questions such as the identification of the heating mechanism, the origin of the observed fast ion tail and the often observed density increase during the HF remain to be investigated. Based on the power balance carried out in WEGA, a first attempt at an explanation of these various effects was carried out. A summary of this work constitutes the first part of this paper. Another very important aspect of lower hybrid heating consists in the possibility of coupling the lower hybrid wave to the plasma by means of a phased waveguide array, the 'grill'. In order to confirm the advantages of the grill structure up to the power density levels necessary in the future, specific experiments have been carried out in the PETULA device. Several results obtained there are presented in the second part of this paper. In the final section the directions of present research necessary in order to ensure applicability to large machines are briefly discussed

  13. Accuracy of HER2 status determination on breast core-needle biopsies (immunohistochemistry, FISH, CISH and SISH vs FISH).

    Science.gov (United States)

    Arnould, Laurent; Roger, Pascal; Macgrogan, Gaëtan; Chenard, Marie-Pierre; Balaton, André; Beauclair, Sophie; Penault-Llorca, Frederique

    2012-05-01

    Preoperative breast cancer diagnosis on core biopsies has become a standard of care in many countries. Controversies exist concerning the accuracy of HER2 testing on biopsies as compared with surgical specimens, and few data exist concerning the use of emerging technologies such as bright-field in-situ hybridization in such a setting. A French multicenter, cross-sectional, histopathological study assessed the concordance of HER2 status determined by immunohistochemistry and silver (SISH) or chromogenic in-situ hybridization (CISH) on core-needle biopsies with HER2 status determined by fluorescence in-situ hybridization (FISH) on surgical specimens. The concordance between biopsy and operative results was also assessed for each method. We studied 260 breast tumors from 24 centers between April 2003 and August 2009. Excellent concordance (κ: 0.92-0.97) was shown between immunohistochemistry and FISH with low discordance rates (2-4%), high specificity (97-98%) and sensitivity values (95-99%), with no significant difference according to the immunohistochemistry interpretation guidelines used. The correlation between SISH and CISH on biopsies and FISH on surgical samples was strong (κ: 0.96 and 0.94, respectively), with no significant difference between false negative rates or sensitivity and specificity values (2 and 5%, 99 and 96%, 98 and 98%, respectively). Whatever the evaluation technique, excellent concordance between biopsies and surgical specimens was observed (κ ≥ 0.97; discordance rates between 1 and 2%), with high sensitivity (98-99%) and specificity (98-100%). Based on these results, when FISH cannot be used, SISH and/or CISH could be proposed as an alternative method to determine HER2 status and to confirm any ambiguous immunohistochemistry results, either for preoperative percutaneous biopsies or for surgical specimens. They could also be used for quality controls and immunohistochemistry calibration.

  14. [HER2 gene amplification assay: is CISH an alternative to FISH?].

    Science.gov (United States)

    Denoux, Yves; Arnould, Laurent; Fiche, Maryse; Lannes, B; Couturier, Jérôme; Vincent-Salomon, Anne; Penault-Llorca, Frédérique; Antoine, M; Balaton, A; Baranzelli, M C; Becette, V; Bellocq, J P; Bibeau, F; Ettore, F; Fridman, V; Gnassia, J P; Jacquemier, J; MacGrogan, G; Mathieu, M C; Migeon, C; Rigaud, C; Roger, P; Sigal-Zafrani, B; Simony-Lafontaine, J; Trassard, M; Treilleux, I; Verriele, V; Voigt, J J

    2003-12-01

    The HER2 proto-oncogene encodes a transmembrane protein, which is considered to function as a growth factor receptor. Overexpression of this protein found by immunohistochemistry in about 20% of infiltrating breast carcinomas, has a predictive value of response to treatment by trastuzumab, an anti-HER2 humanized monoclonal antibody. Search for HER2 gene amplification is necessary to adapt the immunohistochemical technique quality and also in the cases of delicate analysis or weak overexpression. It is usually carried out by Fluorescence In Situ Hybridization (FISH). A more recent hybridization technique, named CISH because of its chromogenic revelation is an alternative method, which gives highly correlated results with FISH. We present details of this technique, which may be more familiar for the pathologists than FISH, because reading analysis is similar to that of immunohistochemical staining.

  15. Temperature effect on gastric emptying time of hybrid grouper (Epinephelus spp.)

    Energy Technology Data Exchange (ETDEWEB)

    De, Moumita; Ghaffar, Mazlan Abd. [School of Environmental and Natural Resource Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor (Malaysia); Das, Simon K. [School of Environmental and Natural Resource Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia and Marine Ecosystem Research Centre (EKOMAR), Faculty of Science and Technology, Universiti (Malaysia)

    2014-09-03

    Knowledge of fish gastric emptying time is a necessary component for understanding the fish feeding rates, energy budgets and commercial production of fishes in aquaculture. The hybrid grouper Epinephelus spp. is getting popular as a culture species in Malaysia for their faster growth rate compared to commonly cultured grouper species (giant grouper Epinephelus lanceolatus and tiger grouper Epinephelus fuscoguttatus). There are data suggests that elevated sea water temperature affects gastric emptying time (GET) of fishes. Hence, this study aims to study the GET of hybrid grouper at different temperature (22, 26, 30, 34°C) in laboratory condition with commercial diet pellet. The gastric emptying times (GETs) at different temperatures were determined X-radiographically, using barium sulfate (BaSO{sub 4}) as a contrast medium food marker. The food marker and X-radiography showed that initial voidance of fecal matter began 4-6 h after feeding at all temperature. The fastest GET (13 h) was obsereved in the 30°C group, whereas the longest (17 h) GET was seen in 22°C group fed with artificial diet pellet. Not much differences in GET were recorded between the 26 and 34°C groups as 34°C groups fed lesser amount compared to 26°C groups. Nevertheless a substantial delay in GET was observed in the 22°C group. The findings of this study suggest to culture hybrid grouper between 26 to 30°C with commercial diet pellet as this temperature ranges proliferate the faster digestion process which may contribute faster growth rate of this commerical important fish species. Overall, these findings may have important consequences for optimization of commercial production of hybrid grouper.

  16. Temperature effect on gastric emptying time of hybrid grouper (Epinephelus spp.)

    Science.gov (United States)

    De, Moumita; Ghaffar, Mazlan Abd.; Das, Simon K.

    2014-09-01

    Knowledge of fish gastric emptying time is a necessary component for understanding the fish feeding rates, energy budgets and commercial production of fishes in aquaculture. The hybrid grouper Epinephelus spp. is getting popular as a culture species in Malaysia for their faster growth rate compared to commonly cultured grouper species (giant grouper Epinephelus lanceolatus and tiger grouper Epinephelus fuscoguttatus). There are data suggests that elevated sea water temperature affects gastric emptying time (GET) of fishes. Hence, this study aims to study the GET of hybrid grouper at different temperature (22, 26, 30, 34°C) in laboratory condition with commercial diet pellet. The gastric emptying times (GETs) at different temperatures were determined X-radiographically, using barium sulfate (BaSO4) as a contrast medium food marker. The food marker and X-radiography showed that initial voidance of fecal matter began 4-6 h after feeding at all temperature. The fastest GET (13 h) was obsereved in the 30°C group, whereas the longest (17 h) GET was seen in 22°C group fed with artificial diet pellet. Not much differences in GET were recorded between the 26 and 34°C groups as 34°C groups fed lesser amount compared to 26°C groups. Nevertheless a substantial delay in GET was observed in the 22°C group. The findings of this study suggest to culture hybrid grouper between 26 to 30°C with commercial diet pellet as this temperature ranges proliferate the faster digestion process which may contribute faster growth rate of this commerical important fish species. Overall, these findings may have important consequences for optimization of commercial production of hybrid grouper.

  17. Temperature effect on gastric emptying time of hybrid grouper (Epinephelus spp.)

    International Nuclear Information System (INIS)

    De, Moumita; Ghaffar, Mazlan Abd.; Das, Simon K.

    2014-01-01

    Knowledge of fish gastric emptying time is a necessary component for understanding the fish feeding rates, energy budgets and commercial production of fishes in aquaculture. The hybrid grouper Epinephelus spp. is getting popular as a culture species in Malaysia for their faster growth rate compared to commonly cultured grouper species (giant grouper Epinephelus lanceolatus and tiger grouper Epinephelus fuscoguttatus). There are data suggests that elevated sea water temperature affects gastric emptying time (GET) of fishes. Hence, this study aims to study the GET of hybrid grouper at different temperature (22, 26, 30, 34°C) in laboratory condition with commercial diet pellet. The gastric emptying times (GETs) at different temperatures were determined X-radiographically, using barium sulfate (BaSO 4 ) as a contrast medium food marker. The food marker and X-radiography showed that initial voidance of fecal matter began 4-6 h after feeding at all temperature. The fastest GET (13 h) was obsereved in the 30°C group, whereas the longest (17 h) GET was seen in 22°C group fed with artificial diet pellet. Not much differences in GET were recorded between the 26 and 34°C groups as 34°C groups fed lesser amount compared to 26°C groups. Nevertheless a substantial delay in GET was observed in the 22°C group. The findings of this study suggest to culture hybrid grouper between 26 to 30°C with commercial diet pellet as this temperature ranges proliferate the faster digestion process which may contribute faster growth rate of this commerical important fish species. Overall, these findings may have important consequences for optimization of commercial production of hybrid grouper

  18. The correlation between dual-color chromogenic in situ hybridization and fluorescence in situ hybridization in assessing HER2 gene amplification in breast cancer.

    Science.gov (United States)

    Pedersen, Marianne; Rasmussen, Birgitte Bruun

    2009-06-01

    Fluorescence in situ hybridization (FISH) is regarded as the gold standard method for detecting HER2 gene amplification. Chromogenic in situ hybridization (CISH) is a promising alternative to FISH because CISH has the advantages of being a method evaluated by bright-field microscopy and the generated chromogenic signals are also stable. This study presents a dual color CISH for simultaneous detection of the HER2 gene and chromosome 17. The CISH method performs a chromogenic detection "on top" of the Food and Drug Administration (FDA)-approved HER2 FISH pharmDx method, where the fluorochrome-labeled probes are detected using enzyme-labeled antibodies and visualized by chromogenic enzymatic reactions. The HER2 status (amplified/not amplified and HER2 ratios) was evaluated by the CISH method and compared with results obtained by the FDA-approved FISH method. Of the 72 successfully investigated invasive breast carcinomas, both FISH and CISH detected HER2 amplification in 24 cases and nonamplification was detected in 47 cases. One case showed a discrepancy between FISH and CISH. The concordance between CISH and FISH was found to be almost perfect (98.6%). The correlation between the HER2 ratios obtained by the 2 methods showed excellent correlation (correlation coefficient 0.95). In conclusion, it is possible by dual-color CISH method to demonstrate HER2 genes and chromosome 17 genes, in the same tissue section and reliably assess HER2 status. The CISH method is a very promising alternative to the FISH method.

  19. Metaphase FISH on a Chip: Miniaturized Microfluidic Device for Fluorescence in situ Hybridization

    DEFF Research Database (Denmark)

    Vedarethinam, Indumathi; Shah, Pranjul Jaykumar; Dimaki, Maria

    2010-01-01

    -FISH, the process continues to be a manual, labour intensive, expensive and time consuming technique, often taking over 3-5 days, even in dedicated labs. We have developed a novel microFISH device to perform metaphase FISH on a chip which overcomes many shortcomings of the current laboratory protocols. This work...... also introduces a novel splashing device for preparing metaphase spreads on a microscope glass slide, followed by a rapid adhesive tape-based bonding protocol leading to rapid fabrication of the microFISH device. The microFISH device allows for an optimized metaphase FISH protocol on a chip with over...... a 20-fold reduction in the reagent volume. This is the first demonstration of metaphase FISH on a microfluidic device and offers a possibility of automation and significant cost reduction of many routine diagnostic tests of genetic anomalies....

  20. Triaxial Accelerometer Error Coefficients Identification with a Novel Artificial Fish Swarm Algorithm

    Directory of Open Access Journals (Sweden)

    Yanbin Gao

    2015-01-01

    Full Text Available Artificial fish swarm algorithm (AFSA is one of the state-of-the-art swarm intelligence techniques, which is widely utilized for optimization purposes. Triaxial accelerometer error coefficients are relatively unstable with the environmental disturbances and aging of the instrument. Therefore, identifying triaxial accelerometer error coefficients accurately and being with lower costs are of great importance to improve the overall performance of triaxial accelerometer-based strapdown inertial navigation system (SINS. In this study, a novel artificial fish swarm algorithm (NAFSA that eliminated the demerits (lack of using artificial fishes’ previous experiences, lack of existing balance between exploration and exploitation, and high computational cost of AFSA is introduced at first. In NAFSA, functional behaviors and overall procedure of AFSA have been improved with some parameters variations. Second, a hybrid accelerometer error coefficients identification algorithm has been proposed based on NAFSA and Monte Carlo simulation (MCS approaches. This combination leads to maximum utilization of the involved approaches for triaxial accelerometer error coefficients identification. Furthermore, the NAFSA-identified coefficients are testified with 24-position verification experiment and triaxial accelerometer-based SINS navigation experiment. The priorities of MCS-NAFSA are compared with that of conventional calibration method and optimal AFSA. Finally, both experiments results demonstrate high efficiency of MCS-NAFSA on triaxial accelerometer error coefficients identification.

  1. Marker chromosome 21 identified by microdissection and FISH

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Y.; Palmer, C.G. [Indiana Univ. School of Medicine, Indianapolis, IN (United States); Rubinstein, J. [Univ. Affiliated Cincinnati Center for Developmental Disorders, OH (United States)] [and others

    1995-03-27

    A child without Down`s syndrome but with developmental delay, short stature, and autistic behavior was found to be mosaic 46,XX/47,XX,+mar(21) de novo. The marker was a small ring or dot-like chromosome. Microdissection of the marker was performed. The dissected fragments were biotinylated with sequence-independent PCR as a probe pool for fluorescence in situ hybridization (FISH). FISH results suggested an acrocentric origin of the marker. Subsequent FISH with {alpha}-satellite DNA probes for acrocentric chromosomes and chromosome-specific 21 and 22 painting probes confirmed its origin from chromosome 21. 14 refs., 3 figs.

  2. THE RESEARCH IN FISH GENETICS IN CROATIA AND FORMER YUGOSLAVIA

    Directory of Open Access Journals (Sweden)

    Tomislav Treer

    1994-03-01

    Full Text Available This is a review on fish genetics research in Croatia and former Yugoslavia, based on the analyses of all the articles published in four main journals (Ribarstvo Jugoslavije, Morsko ribarstvo, Ichthyologia and Acta Adriatica since 1945 till disintegration of Yugoslavia in 1991. Most of the papers cover the fields on cytogenetics and hybridization (24 and 13 respectively. Eight papers were on fish selection and five on population genetics. Apart from those, five papers were written by foreign authors. Two groups of researchers from the University of Sarajevo were specially active. One of them lead by B e r b e r o v i ć and S o f r a d ž i j a did extensive work in cytogenetics, analyzing the karyotypes of many fish species, some of them endemic. Another one lead by V u k o v i ć , investigated some natural hybrids and created many of them artificially, particulary among cyprinids. These results are presented in a special table. Contrary to the mountainous Bosnia where this type of research was of systematic and ecologic importance, in Croatia whwrw aquaculture was highly developed, the approach was quite different. The scientists from the University of Zagreb, H a b e k o v i ć and T u r k , studied the hybridization and selection of important cultured cyprinids. Apart from these scientific groups, many papers were published by A l - S a b t i , who later became world famous in fish cytogenetics. The works of many other authors who contributed with papers in different fields of fish genetics are also described.

  3. Hybrid rocket engine, theoretical model and experiment

    Science.gov (United States)

    Chelaru, Teodor-Viorel; Mingireanu, Florin

    2011-06-01

    The purpose of this paper is to build a theoretical model for the hybrid rocket engine/motor and to validate it using experimental results. The work approaches the main problems of the hybrid motor: the scalability, the stability/controllability of the operating parameters and the increasing of the solid fuel regression rate. At first, we focus on theoretical models for hybrid rocket motor and compare the results with already available experimental data from various research groups. A primary computation model is presented together with results from a numerical algorithm based on a computational model. We present theoretical predictions for several commercial hybrid rocket motors, having different scales and compare them with experimental measurements of those hybrid rocket motors. Next the paper focuses on tribrid rocket motor concept, which by supplementary liquid fuel injection can improve the thrust controllability. A complementary computation model is also presented to estimate regression rate increase of solid fuel doped with oxidizer. Finally, the stability of the hybrid rocket motor is investigated using Liapunov theory. Stability coefficients obtained are dependent on burning parameters while the stability and command matrixes are identified. The paper presents thoroughly the input data of the model, which ensures the reproducibility of the numerical results by independent researchers.

  4. Hybrid Reynolds-Averaged/Large-Eddy Simulations of a Coaxial Supersonic Free-Jet Experiment

    Science.gov (United States)

    Baurle, Robert A.; Edwards, Jack R.

    2010-01-01

    Reynolds-averaged and hybrid Reynolds-averaged/large-eddy simulations have been applied to a supersonic coaxial jet flow experiment. The experiment was designed to study compressible mixing flow phenomenon under conditions that are representative of those encountered in scramjet combustors. The experiment utilized either helium or argon as the inner jet nozzle fluid, and the outer jet nozzle fluid consisted of laboratory air. The inner and outer nozzles were designed and operated to produce nearly pressure-matched Mach 1.8 flow conditions at the jet exit. The purpose of the computational effort was to assess the state-of-the-art for each modeling approach, and to use the hybrid Reynolds-averaged/large-eddy simulations to gather insight into the deficiencies of the Reynolds-averaged closure models. The Reynolds-averaged simulations displayed a strong sensitivity to choice of turbulent Schmidt number. The initial value chosen for this parameter resulted in an over-prediction of the mixing layer spreading rate for the helium case, but the opposite trend was observed when argon was used as the injectant. A larger turbulent Schmidt number greatly improved the comparison of the results with measurements for the helium simulations, but variations in the Schmidt number did not improve the argon comparisons. The hybrid Reynolds-averaged/large-eddy simulations also over-predicted the mixing layer spreading rate for the helium case, while under-predicting the rate of mixing when argon was used as the injectant. The primary reason conjectured for the discrepancy between the hybrid simulation results and the measurements centered around issues related to the transition from a Reynolds-averaged state to one with resolved turbulent content. Improvements to the inflow conditions were suggested as a remedy to this dilemma. Second-order turbulence statistics were also compared to their modeled Reynolds-averaged counterparts to evaluate the effectiveness of common turbulence closure

  5. Fluorescence In Situ Hybridization (FISH-Based Karyotyping Reveals Rapid Evolution of Centromeric and Subtelomeric Repeats in Common Bean (Phaseolus vulgaris and Relatives

    Directory of Open Access Journals (Sweden)

    Aiko Iwata-Otsubo

    2016-04-01

    Full Text Available Fluorescence in situ hybridization (FISH-based karyotyping is a powerful cytogenetics tool to study chromosome organization, behavior, and chromosome evolution. Here, we developed a FISH-based karyotyping system using a probe mixture comprised of centromeric and subtelomeric satellite repeats, 5S rDNA, and chromosome-specific BAC clones in common bean, which enables one to unambiguously distinguish all 11 chromosome pairs. Furthermore, we applied the karyotyping system to several wild relatives and landraces of common bean from two distinct gene pools, as well as other related Phaseolus species, to investigate repeat evolution in the genus Phaseolus. Comparison of karyotype maps within common bean indicates that chromosomal distribution of the centromeric and subtelomeric satellite repeats is stable, whereas the copy number of the repeats was variable, indicating rapid amplification/reduction of the repeats in specific genomic regions. In Phaseolus species that diverged approximately 2–4 million yr ago, copy numbers of centromeric repeats were largely reduced or diverged, and chromosomal distributions have changed, suggesting rapid evolution of centromeric repeats. We also detected variation in the distribution pattern of subtelomeric repeats in Phaseolus species. The FISH-based karyotyping system revealed that satellite repeats are actively and rapidly evolving, forming genomic features unique to individual common bean accessions and Phaseolus species.

  6. [Influences of ration level and initial body weight on growth and size hierarchy of hybrid tilapia, Oreochromis mossambicus x O. niloticus].

    Science.gov (United States)

    Wang, Yan

    2003-02-01

    72 hybrid tilapia were distributed into twelve tanks, each stocked in six fish (A, B, C, D, E, F) with different body size. The initial body weight was 62.69 +/- 1.46 g for fish A, 56.48 +/- 1.30 g for B, 50.75 +/- 1.19 g for C, 35.56 +/- 1.18 g for D, 31.05 +/- 0.88 g for E, and 27.35 +/- 0.95 g for F (mean +/- SE). The fish were reared under four ration levels (deprived of feed, fed at 1.5% body weight per day (BW.d-1), fed at 3.0% BW.d-1, or fed excess) throughout four weeks, respectively. The specific growth rate (SGR) and feed efficiency (FE) of the fish increased with increasing ration level up to 3.0% BW.d-1, and then, SGR maintained at a constant level while FE dropped with continuing increase of ration. The variance coefficient of SGR and final body weight was relatively high in the fish that fed at 1.5% BW.d-1. For fish A, SGR was not significantly different, between 1.5% BW.d-1, 3.0% BW.d-1 and fed excess, while in fish F, relatively high variations were found among different ration treatments. This experiment revealed that the growth and size hierarchy of hybrid tilapia were related to their ration level and initial body size. Under ration levels more than 3.0% BW.d-1, SGR was usually high, and size hierarchy was relatively weak.

  7. Electromagnetic radiation by parametric decay of upper hybrid waves in ionospheric modification experiments

    International Nuclear Information System (INIS)

    Leyser, T.B.

    1994-01-01

    A nonlinear dispersion relation for the parametric decay of an electrostatic upper hybrid wave into an ordinary mode electromagnetic wave, propagating parallel to the ambient magnetic field, and an electrostatic low frequency wave, being either a lower hybrid wave or a high harmonic ion Bernstein wave, is derived. The coherent and resonant wave interaction is considered to take place in a weakly magnetized and collisionless Vlasov plasma. The instability growth rate is computed for parameter values typical of ionospheric modification experiments, in which a powerful high frequency electromagnetic pump wave is injected into the ionospheric F-region from ground-based transmitters. The electromagnetic radiation which is excited by the decaying upper hybrid wave is found to be consistent with the prominent and commonly observed downshifted maximum (DM) emission in the spectrum of stimulated electromagnetic emission

  8. mathFISH, a Web Tool That Uses Thermodynamics-Based Mathematical Models for In Silico Evaluation of Oligonucleotide Probes for Fluorescence In Situ Hybridization▿ †

    OpenAIRE

    Yilmaz, L. Safak; Parnerkar, Shreyas; Noguera, Daniel R.

    2010-01-01

    Mathematical models of RNA-targeted fluorescence in situ hybridization (FISH) for perfectly matched and mismatched probe/target pairs are organized and automated in web-based mathFISH (http://mathfish.cee.wisc.edu). Offering the users up-to-date knowledge of hybridization thermodynamics within a theoretical framework, mathFISH is expected to maximize the probability of success during oligonucleotide probe design.

  9. Initial experience in hybrid PET-MRI for evaluation of refractory focal onset epilepsy.

    Science.gov (United States)

    Shin, Hae W; Jewells, Valerie; Sheikh, Arif; Zhang, Jingwen; Zhu, Hongtu; An, Hongyu; Gao, Wei; Shen, Dinggang; Hadar, Eldad; Lin, Weili

    2015-09-01

    We aim to evaluate the utility/improved accuracy of hybrid PET/MR compared to current practice separate 3T MRI and PET-CT imaging for localization of seizure foci. In a pilot study, twenty-nine patients undergoing epilepsy surgery evaluation were imaged using PET/MR. This subject group had 29 previous clinical 3T MRI as well as 12 PET-CT studies. Prior clinical PET and MR images were read sequentially while the hybrid PET/MR was concurrently read. The median interval between hybrid PET/MR and prior imaging studies was 5 months (range 1-77 months). In 24 patients, there was no change in the read between the clinical exams and hybrid PET/MR while new anatomical or functional lesions were identified by hybrid PET/MR in 5 patients without significant clinical change. Four new anatomical MR lesions were seen with concordant PET findings. The remaining patient revealed a new abnormal PET lesion without an MR abnormality. All new PET/MR lesions were clinically significant with concordant EEG and/or SPECT results as potential epileptic foci. Our initial hybrid PET-MRI experience increased diagnostic yields for detection of potential epileptic lesions. This may be due to the unique advantage of improved co-registration and simultaneous review of both structural and functional data. Copyright © 2015 British Epilepsy Association. Published by Elsevier Ltd. All rights reserved.

  10. Towards Fluorescence In Vivo Hybridization (FIVH) Detection of H. pylori in Gastric Mucosa Using Advanced LNA Probes

    DEFF Research Database (Denmark)

    Fontenete, Sílvia; Leite, Marina; Guimarães, Nuno

    2015-01-01

    acid (LNA)/ 2' O-methyl RNA (2'OMe) probe using standard phosphoramidite chemistry and FISH hybridization was then successfully performed both on adhered and suspended bacteria at 37°C. In this work we simplified, shortened and adapted FISH to work at gastric pH values, meaning that the hybridization...... step now takes only 30 minutes and, in addition to the buffer, uses only urea and probe at non-toxic concentrations. Importantly, the sensitivity and specificity of the FISH method was maintained in the range of conditions tested, even at low stringency conditions (e.g., low pH). In conclusion......In recent years, there have been several attempts to improve the diagnosis of infection caused by Helicobacter pylori. Fluorescence in situ hybridization (FISH) is a commonly used technique to detect H. pylori infection but it requires biopsies from the stomach. Thus, the development of an in vivo...

  11. Expanding diversity of potential bacterial partners of the methanotrophic ANME archaea using Magneto-FISH

    Science.gov (United States)

    Trembath-Reichert, E.; Green-Saxena, A.; Steele, J. A.; Orphan, V. J.

    2012-12-01

    Sulfate-coupled anaerobic oxidation of methane (AOM) in marine sediments is the major sink for methane in the oceans. This process is believed to be catalyzed by as yet uncultured syntrophic consortia of ANME archaea (affiliated with the Methanosarcinales) and sulfate-reducing bacteria belonging to the Desulfosarcina/Desulfococcus and Desulfobulbaceae. These syntrophic consortia have been described from methane-rich habitats worldwide and appear to be most concentrated in areas of high methane flux, such as cold seeps along continental margins. The extent of the diversity and ecophysiological potential of these microbial associations is still poorly constrained. In an effort to better characterize the diversity of microorganisms forming associations with different clades of methanotrophic ANME archaea (ANME-1, ANME-2a/b/c, ANME-3) and link these organisms to potentially diagnostic metabolic genes (e.g. mcrA, dsrAB, aprA), we employed a unique culture-independent whole cell capture technique which combines Fluorescence In Situ Hybridization with immuno-magnetic cell capture (Magneto-FISH). We used Magneto-FISH for targeted enrichment of specific ANME groups and their associated bacteria directly from formalin- and ethanol-fixed methane seep sediment. The identity and metabolic gene diversity of captured microorganisms were then assessed by clone library construction and sequencing. Diversity recovered from Magneto-FISH experiments using general and clade-specific ANME targeted probes show both the expected selectivity of the FISH probes (i.e. predominately ANME-2c subclade captured with an ANME-2c probe and multiple ANME groups recovered with the general probe targeting most ANME). Follow up FISH experiments were conducted to confirm physical associations between ANME and unique bacterial members (deltaproteobacteria and other non-sulfate reducing groups) that were common to multiple Magneto-FISH capture experiments. Analyses of metabolic gene diversity for archaeal

  12. TNB Experience in Developing Solar Hybrid Station at RPS Kemar, Gerik, Perak Darul Ridzuan

    International Nuclear Information System (INIS)

    Aziz, K A; Shamsudin, K N

    2013-01-01

    This paper will discuss on TNB experience in developing Solar Hybrid Station at RPS Kemar, Gerik, Perak. TNB has been approached by KKLW to submit proposal to provide electricity in the rural area namely RPS Kemar. Looking at area and source available, Solar Hybrid System was the best method in order to provide electricity at this area. This area is far from national grid sources. Solar Hybrid System is the best method to produce electrical power using the renewable energy from Solar PV, Battery and Diesel Generator Set. Nowadays, price of petroleum is slightly high due to higher demand from industry. Solar energy is good alternative in this country to practice in order to reduce cost for produce of electrical energy. Generally, Solar will produce energy during daytime and when become cloudy and dark, automatically battery and diesel generator set will recover the system through the hybrid controller system.

  13. EGFR status in oral squamous cell carcinoma: comparing immunohistochemistry, FISH and CISH detection in a case series study

    OpenAIRE

    Bernardes, Vanessa F?tima; Gleber-Netto, Frederico Omar; de Sousa, S?lvia Ferreira; Rocha, Rafael Malagoli; de Aguiar, Maria C?ssia Ferreira

    2013-01-01

    Objectives To compare the immunohistochemistry (IHC) expression of epidermal growth factor receptor (EGFR) in oral squamous cell carcinomas (OSCC) with the gene amplification evaluated by fluorescence in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) and their association with clinicopathological parameters. Additionally, we tested the sensibility and specificity of CISH in comparison with FISH. Design Case series study Setting Oral surgery and pathology department in ...

  14. A novel, simple and rapid nondenaturing FISH (ND-FISH) technique for the detection of plant telomeres. Potential used and possible target structures detected.

    Science.gov (United States)

    Cuadrado, Angeles; Golczyk, Hieronim; Jouve, Nicolás

    2009-01-01

    We report a new technique-nondenaturing FISH (ND-FISH)-for the rapid detection of plant telomeres without the need for prior denaturation of the chromosomes. In its development, two modified, synthetic oligonucleotides, 21 nt in length, fluorescently labelled at their 5' and 3' ends and complementary to either the cytidine-rich (C(3)TA(3)) or guanosine-rich (T(3)AG(3)) telomeric DNA strands, were used as probes. The high binding affinity of these probes and the short hybridization time required allows the visualization of plant telomeres in less than an hour. In tests, both probes gave strong signals visualized as double spots at both chromosome ends; this was true of both the mitotic and meiotic chromosomes of barley, wheat, rye, maize, Brachypodium distachyon and Rhoeo spathacea. They were also able to detect telomere motifs at certain intercalary sites in the chromosomes of R. spathacea. To investigate the nature of the target structures detected, the chromosomes were treated with RNase A and single strand-specific nuclease S1 before ND-FISH experiments. Signal formation was resistant to standard enzymatic treatment, but sensitive when much higher enzyme concentrations were used. The results are discussed in relation to current knowledge of telomere structure.

  15. Reliability of chromogenic in situ hybridization for epidermal growth factor receptor gene copy number detection in non-small-cell lung carcinomas: a comparison with fluorescence in situ hybridization study.

    Science.gov (United States)

    Yoo, Seol Bong; Lee, Hyun Ju; Park, Jung Ok; Choe, Gheeyoung; Chung, Doo Hyun; Seo, Jeong-Wook; Chung, Jin-Haeng

    2010-03-01

    Fluorescence in situ hybridization (FISH) has been known to be the most representative and standardized test for assessing gene amplification. However, FISH requires a fluorescence microscope, the signals are labile and rapidly fade over time. Recently, chromogenic in situ hybridization (CISH) has emerged as a potential alternative to FISH. The aim of this study is to test the reliability of CISH technique for the detection of epidermal growth factor receptor (EGFR) gene amplification in non-small-cell lung carcinomas (NSCLC), to compare CISH results with FISH. A total of 277 formalin-fixed and paraffin embedded NSCLC tissue samples were retrieved from the surgical pathology archives at Seoul National University Bundang Hospital. CISH and FISH examinations were performed to test EGFR gene amplification status. There was high concordance in the assessment of EGFR gene copy number between CISH and FISH tests (Kappa coefficient=0.83). Excellent concordance was shown between two observers on the interpretation of the CISH results (Kappa coefficient=0.90). In conclusion, CISH result is highly reproducible, accurate and practical method to determine EGFR gene amplification in NSCLC. In addition, CISH allows a concurrent analysis of histological features of the tumors and gene copy numbers.

  16. In situ hybridization; principles and applications: review article

    Directory of Open Access Journals (Sweden)

    Zahra Nozhat

    2015-06-01

    Full Text Available In situ hybridization (ISH is a method that uses labeled complementary single strand DNA or RNA to localize specific DNA or RNA sequences in an intact cell or in a fixed tissue section. The main steps of ISH consist of: probe selection, tissue or sample preparation, pre-hybridization treatment, hybridization and washing, detection and control procedure. Probe selection is one of the important aspects of successful hybridization. ISH sensitivity and specificity can be influenced by: probe construct, efficiency of labeling, percentage of GC, probe length and signal detection systems. Different methods such as nick translation, random priming, end tailing and T4 DNA polymerase replacement are used for probe generation. Both radioactive and non-radioactive labels can be used in order to probe labeling. Nucleic acid maintenance in samples, prevention of morphological changes of samples and probe penetration into tissue section are the main aims of sample preparation step. Then, a small amount of solution containing probe, is added on slides containing tissue sections for hybridization process, then slides are incubated overnight. Next day, washes are carried out to remove the probes which are not bound to target DNA or RNA. Finally, in order to be sure that the observed labeling is specific to the target sequence, using several control procedures is very important. Various techniques based on ISH consist of: Fluorescence in situ hybridization (FISH, chromogenic in situ hybridization (CISH, genomic in situ hybridization (GISH, comparative genomic hybridization (CGH, spectral karyotyping (SKY and multiplex fluorescence in situ hybridization (MFISH. One of the most common techniques of ISH is fluorescence in situ hybridization. FISH can be used to: 1 detect small deletions and duplications that are not visible using microscope analysis, 2 detect how many chromosomes of a certain type are present in each cell and 3 confirm rearrangements that are

  17. Stationary magnetic shear reversal during Lower Hybrid experiments in Tore Supra

    International Nuclear Information System (INIS)

    Litaudon, X.; Arslanbekov, R.; Hoang, G.T.; Joffrin, E.; Kazarian-Vibert, F.; Moreau, D.; Peysson, Y.; Bibet, P.

    1996-01-01

    Stable and stationary states with hollow current density profiles have been achieved with Lower Hybrid Current Drive (LHCD) during Lower Hybrid (LH) wave accessibility experiments. By analysing the bounded propagation domain in phase space which naturally limits the central penetration and absorption of the waves, off-axis LH power deposition has been realized in a reproducible manner. The resulting current density profile modifications have led to a global confinement enhancement attributed to the formation of an internal 'transport barrier' in the central reversed shear region where the electron thermal diffusivity is reduced to its neoclassical collisional level. The multiple-pass LH wave propagation in the weak Landau damping and reversed magnetic shear regime is also investigated in the framework of a statistical theory and the experimental validation of this theory is discussed. (author)

  18. MOLECULAR GENETIC MARKERS AND METHODS OF THEIR IDENTIFICATION IN MODERN FISH-FARMING

    Directory of Open Access Journals (Sweden)

    I. Hrytsyniak

    2014-03-01

    Full Text Available Purpose. The application of molecular genetic markers has been widely used in modern experimental fish-farming in recent years. This methodology is currently presented by a differentiated approach with individual mechanisms and clearly defined possibilities. Numerous publications in the scientific literature that are dedicated to molecular genetic markers for the most part offer purely practical data. Thus, the synthesis and analysis of existing information on the general principles of action and the limits of the main methods of using molecular genetic markers is an actual problem. In particular, such a description will make it possible to plan more effectively the experiment and to obtain the desired results with high reliability. Findings. The main types of variable parts of DNA that can be used as molecular genetic markers in determining the level of stock hybridization, conducting genetic inventory of population and solving other problems in modern fish-farming are described in this paper. Also, the article provides an overview of principal modern methods that can be used to identify molecular genetic markers. Originality. This work is a generalization of modern ideas about the mechanisms of experiments with molecular genetic markers in fish-farming. Information is provided in the form of consistent presentation of the principles and purpose of each method, as well as significant advances during their practical application. Practical value. The proposed review of classic and modern literature data on molecular genetic markers can be used for planning, modernization and correction of research activity in modern fish-farming.

  19. Fluorescent in situ hybridization (FISH) on corneal impression cytology specimens (CICS): study of epithelial cell survival after keratoplasty.

    Science.gov (United States)

    Catanese, Muriel; Popovici, Cornel; Proust, Hélène; Hoffart, Louis; Matonti, Frédéric; Cochereau, Isabelle; Conrath, John; Gabison, Eric E

    2011-02-22

    To assess corneal epithelial cell survival after keratoplasty. Corneal impression cytology (CIC) was performed on sex-mismatched corneal transplants. Fluorescent in situ hybridization (FISH) with sex chromosome-specific probes was performed to identify epithelial cell mosaicism and therefore allocate the donor or recipient origin of the cells. Twenty-four samples of corneal epithelial cells derived from 21 transplanted patients were analyzed. All patients received post-operative treatment using dexamethasone eye drops, with progressive tapering over 18 months, and nine patients also received 2% cyclosporine eye drops. Out of the 24 samples reaching quality criteria, sex mosaicism was found in 13, demonstrating the presence of donor-derived cells at the center of the graft for at least 211 days post keratoplasty. Kaplan-Meier analysis established a median survival of donor corneal epithelial cells of 385 days. Although not statistically significant, the disappearance of donor cells seemed to be delayed and the average number of persistent cells appeared to be greater when 2% cyclosporine was used topically as an additional immunosuppressive therapy. The combination of corneal impressions and FISH analysis is a valuable tool with negligible side effects to investigate the presence of epithelial cell mosaicism in sex-mismatched donor transplants. Epithelial cells survived at the center of the graft with a median survival of more than one year, suggesting slower epithelial turnover than previously described.

  20. Effect of Stocking Density on Growth Performance of Hybrids of ...

    African Journals Online (AJOL)

    Fingerlings of Oreochromis niloticus♀ and Oreochromis urolepis urolepis♂ hybrids were reared at stocking densities of control, 5, 10, 15 and 20 fish/m3 at 15 Practical Salinity Units (PSU) in 1m3 plastic tanks for 63 days. They were kept at low, intermediate and high densities respectively. All hybrids were fed on a ...

  1. 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…

  2. Principle, function, experiences, hybrid chilled ceilings; Prinzip, Funktion, Erfahrungen. Hybrid-Kuehldecken

    Energy Technology Data Exchange (ETDEWEB)

    Schulz, Uwe W. [Hochschule Luzern (Switzerland). Technik und Architektur

    2010-12-15

    Hybrid chilled ceilings combine a radiation chilled ceiling with thermal activations of a concrete ceiling. The activation permits the utilization of alternative and/or small cold generators for the efficient night cooling. The traditional chilled ceiling grants a speedy reaction to load variations as well as a reduction of the reverberation periods. In addition, the combination supplies a draught-free insertion of air. Beside established solutions, MWH Barcol-Air (Staefa, Switzerland) also offers project-specific hybrid chilled ceilings.

  3. Dispersal and selection mediate hybridization between a native and invasive species

    Science.gov (United States)

    Kovach, Ryan P.; Muhlfeld, Clint C.; Boyer, Matthew C.; Lowe, Winsor H.; Allendorf, Fred W.; Luikart, Gordon

    2015-01-01

    Hybridization between native and non-native species has serious biological consequences, but our understanding of how dispersal and selection interact to influence invasive hybridization is limited. Here, we document the spread of genetic introgression between a native (Oncorhynchus clarkii) and invasive (Oncorhynchus mykiss) trout, and identify the mechanisms influencing genetic admixture. In two populations inhabiting contrasting environments, non-native admixture increased rapidly from 1984 to 2007 and was driven by surprisingly consistent processes. Individual admixture was related to two phenotypic traits associated with fitness: size at spawning and age of juvenile emigration. Fish with higher non-native admixture were larger and tended to emigrate at a younger age—relationships that are expected to confer fitness advantages to hybrid individuals. However, strong selection against non-native admixture was evident across streams and cohorts (mean selection coefficient against genotypes with non-native alleles (s) ¼ 0.60; s.e. ¼ 0.10). Nevertheless, hybridization was promoted in both streams by the continuous immigration of individuals with high levels of non-native admixture from other hybrid source populations. Thus, antagonistic relationships between dispersal and selection are mediating invasive hybridization between these fish, emphasizing that data on dispersal and natural selection are needed to fully understand the dynamics of introgression between native and non-native species. .

  4. Double hypernuclei experiment with hybrid emulsion method at J-PARC

    International Nuclear Information System (INIS)

    Ekawa, Hiroyuki

    2015-01-01

    Double hypernuclei are important probes to study the system with strangeness S = -2. Several emulsion experiments had been performed to search for them. We are planning a new experiment to search for double hypernuclei at the K1.8 beam line in the Hadron Experimental Facility (J-PARC E07 experiment). Ξ"- tracks in the emulsion plates and SSD will be automatically connected by a hybrid method. The estimated Ξ"- stopped statistics is 10 times as high as that of the KEK E373 experiment. Discoveries of 10 new double hypernuclear species are expected, which enable us to discuss binding energy in terms of mass number dependence. On the other hand, we will also observe X rays from Ξ"- atoms with a germanium detector array installed close to the emulsion plates by tagging Ξ"- stopped events. This will be the first measurement to give information on the Ξ"- potential at the nuclear surface region. (author)

  5. Fluorescence in situ hybridization of old G-banded and mounted chromosome preparations

    DEFF Research Database (Denmark)

    Gerdes, A M; Pandis, N; Bomme, L

    1997-01-01

    the coverslips detach spontaneously; any mechanical manipulation will jeopardize the results. The success of chromosome painting is improved by excluding the regular RNase treatment step prior to hybridization. Additional changes compared with standard FISH protocols are that the 2 x SSC step is omitted......An improved method for fluorescence in situ hybridization (FISH) investigation of old, previously G-banded, mounted chromosome preparations with chromosome specific painting probes and centromere-specific probes is described. Before hybridization, the slides are incubated in xylene until......, that the amount of added probe is increased approximately 2.5 times, and that the amplification of signals is performed twice. The applicability of the method, which allows double painting with two differently labeled probes using two differently fluorescing colors, was tested on 11 cases involving different...

  6. Enhanced lower hybrid current drive experiments on HT-7 tokamak

    International Nuclear Information System (INIS)

    Shen Weici; Kuang Guangli; Liu Yuexiu; Ding Bojiang; Shi Yaojiang

    2003-01-01

    Effective Lower Hybrid Current Driving (LHCD) and improved confinement experiments in higher plasma parameters (I p >200 kA, n e >2 x 10 13 cm -3 , T e ≥1 keV) have been curried out in optimized LH wave spectrum and plasma parameters in HT-7 superconducting tokamak. The dependence of current driving efficiency on LH power spectrum, plasma density (anti n e ) and toroidal magnetic field B T has been obtained under optimal conditions. A good CD efficiency was obtained at higher plasma current and higher electron density. The improvement of the energy confinement time is accompanied with the increase in line averaged electron density, and in ion and electron temperatures. The highest current driving efficiency reached η CD =I p (anti n e )R/P RF ≅1.05 x 10 19 Am -2 /W. Wave-plasma coupling was sustained in a good state and the reflective coefficient was less than 5%. The experiments have also demonstrated the ability of LH wave in the start-up and ramp-up of the plasma current. The measurement of the temporal distribution of plasma parameter shows that lower hybrid leads to a broader profile in plasma parameter. The LH power deposition profile and the plasma current density profile were modeled with a 2D Fokker-Planck code corresponding to the evolution process of the hard x-ray detector array

  7. Classification and coding of commercial fishing injuries by work processes: an experience in the Danish fresh market fishing industry

    DEFF Research Database (Denmark)

    Jensen, Olaf Chresten; Stage, Søren; Noer, Preben

    2005-01-01

    BACKGROUND: Work-related injuries in commercial fishing are of concern internationally. To better identify the causes of injury, this study coded occupational injuries by working processes in commercial fishing for fresh market fish. METHODS: A classification system of the work processes was deve......BACKGROUND: Work-related injuries in commercial fishing are of concern internationally. To better identify the causes of injury, this study coded occupational injuries by working processes in commercial fishing for fresh market fish. METHODS: A classification system of the work processes...... to working with the gear and nets vary greatly in the different fishing methods. Coding of the injuries to the specific working processes allows for targeted prevention efforts....

  8. Australian experience of providing for fish passage at small instream structures

    Energy Technology Data Exchange (ETDEWEB)

    Lewis, B. [Forest Hill, Victoria (Australia)

    2006-07-01

    Various instream structures have been constructed in Australia as a result of increasing agricultural activities. However, even small structures such as culverts and stream gauging stations can restrict essential fish movements and result in the extinction of local fish species. This paper discussed methods of modifying and designing new structures to ensure adequate fish passage. It was suggested that instream structures can provide for fish passage through the provision of bridges, or through the use of low profile structures for small weirs. Recommendations for site-specific instream structures included an assessment of fish species, topography, flow characteristics and cost effectiveness. Solutions for reducing the impact of small instream barriers to fish movement were also discussed. Provision for fish passage is an important consideration for planners and designers of dams. Legislation is now in place to ensure a planning and approval process prior to the commencement of construction and operation. It was concluded that significant works are now being undertaken to restore fish migration pathways caused by barriers that restrict fish movement. However, monitoring is needed to ensure that designs operate effectively. 17 refs., 3 figs.

  9. Australian experience of providing for fish passage at small instream structures

    International Nuclear Information System (INIS)

    Lewis, B.

    2006-01-01

    Various instream structures have been constructed in Australia as a result of increasing agricultural activities. However, even small structures such as culverts and stream gauging stations can restrict essential fish movements and result in the extinction of local fish species. This paper discussed methods of modifying and designing new structures to ensure adequate fish passage. It was suggested that instream structures can provide for fish passage through the provision of bridges, or through the use of low profile structures for small weirs. Recommendations for site-specific instream structures included an assessment of fish species, topography, flow characteristics and cost effectiveness. Solutions for reducing the impact of small instream barriers to fish movement were also discussed. Provision for fish passage is an important consideration for planners and designers of dams. Legislation is now in place to ensure a planning and approval process prior to the commencement of construction and operation. It was concluded that significant works are now being undertaken to restore fish migration pathways caused by barriers that restrict fish movement. However, monitoring is needed to ensure that designs operate effectively. 17 refs., 3 figs

  10. CARD-FISH for Environmental Microorganisms: Technical Advancement and Future Applications

    OpenAIRE

    Kubota, Kengo

    2012-01-01

    Fluorescence in situ hybridization (FISH) has become a standard technique in environmental microbiology. More than 20 years have passed since this technique was first described, and it is currently used for the detection of ribosomal RNA, messenger RNA, and functional genes encoded on chromosomes. This review focuses on the advancement and applications of FISH combined with catalyzed reporter deposition (CARD, also known as tyramide signal amplification or TSA), in the detection of environmen...

  11. Lower hybrid experiments in the PETULA Tokamak

    International Nuclear Information System (INIS)

    Melin, G.; Bardet, R.; Bernard, M.

    1979-01-01

    The present paper deals with the application of high RF power near the lower hybrid (LH) frequency to a Tokamak discharge. A double waveguide grill antenna has been used to launch a 1.25 GHz-500kW-6 ms RF pulse in PETULA. One of the objectives of this study is to investigate the technological problems associated with LH heating and caused by the high level of RF power involved. Two aspects have been carefully studied: grill conditioning and grill coupling. The latter is strongly dependent on the scrape-off layer of the PETULA plasma and the observations are possibly not significant. The former aspect though not really understood is probably to be expected in other LH experiments. Plasma experiments have been carried out with a wide range of electron density 2. 13 cm -3 in both hydrogen and deuterium. For hydrogen this allows theoretically linear mode conversion to take place in the discharge. Although some heating has been observed, the experiments are not, as yet, very conclusive. The actual RF pulse duration is hardly larger than the ion energy confinement time, and the radial plasma displacement caused by the RF is not feedback controlled. The main discharge parameters were the following: plasma radius 14 cm, toroidal magnetic field 27 kG, plasma current 75 kA, loop voltage 1.6 V, Zsub(eff) < 2., 800 < Tsub(e)(o)< 1100eV, 200 < Tsub(i)(o)< 350 eV

  12. Shoal bass hybridization in the Chattahoochee River Basin near Atlanta, Georgia

    Science.gov (United States)

    Taylor, Andrew T.; Tringali, Michael D.; O'Rourke, Patrick M.; Long, James M.

    2018-01-01

    The shoal bass (Micropterus cataractae) is a sportfish endemic to the Apalachicola-Chattahoochee-Flint Basin of the southeastern United States. Introgression with several non-native congeners poses a pertinent threat to shoal bass conservation, particularly in the altered habitats of the Chattahoochee River. Our primary objective was to characterize hybridization in shoal bass populations near Atlanta, Georgia, including a population inhabiting Big Creek and another in the main stem Chattahoochee River below Morgan Falls Dam (MFD). A secondary objective was to examine the accuracy of phenotypic identifications below MFD based on a simplified suite of characters examined in the field. Fish were genotyped with 16 microsatellite DNA markers, and results demonstrated that at least four black bass species were involved in introgressive hybridization. Of 62 fish genotyped from Big Creek, 27% were pure shoal bass and 65% represented either F1 hybrids of shoal bass x smallmouth bass (M. dolomieu) or unidirectional backcrosses towards shoal bass. Of 29 fish genotyped below MFD and downstream at Cochran Shoals, 45% were pure shoal bass. Six hybrid shoal bass included both F1 hybrids and backcrosses with non-natives including Alabama bass (M. henshalli), spotted bass (M. punctulatus), and smallmouth bass. Shoal bass alleles comprised only 21% of the overall genomic composition in Big Creek and 31% below MFD (when combined with Cochran Shoals). Phenotypic identification below MFD resulted in an overall correct classification rate of 86% when discerning pure shoal bass from all other non-natives and hybrids. Results suggest that although these two shoal bass populations feature some of the highest introgression rates documented, only a fleeting opportunity may exist to conserve pure shoal bass in both populations. Continued supplemental stocking of pure shoal bass below MFD appears warranted to thwart increased admixture among multiple black bass taxa, and a similar stocking

  13. Hybrid Reynolds-Averaged/Large-Eddy Simulations of a Co-Axial Supersonic Free-Jet Experiment

    Science.gov (United States)

    Baurle, R. A.; Edwards, J. R.

    2009-01-01

    Reynolds-averaged and hybrid Reynolds-averaged/large-eddy simulations have been applied to a supersonic coaxial jet flow experiment. The experiment utilized either helium or argon as the inner jet nozzle fluid, and the outer jet nozzle fluid consisted of laboratory air. The inner and outer nozzles were designed and operated to produce nearly pressure-matched Mach 1.8 flow conditions at the jet exit. The purpose of the computational effort was to assess the state-of-the-art for each modeling approach, and to use the hybrid Reynolds-averaged/large-eddy simulations to gather insight into the deficiencies of the Reynolds-averaged closure models. The Reynolds-averaged simulations displayed a strong sensitivity to choice of turbulent Schmidt number. The baseline value chosen for this parameter resulted in an over-prediction of the mixing layer spreading rate for the helium case, but the opposite trend was noted when argon was used as the injectant. A larger turbulent Schmidt number greatly improved the comparison of the results with measurements for the helium simulations, but variations in the Schmidt number did not improve the argon comparisons. The hybrid simulation results showed the same trends as the baseline Reynolds-averaged predictions. The primary reason conjectured for the discrepancy between the hybrid simulation results and the measurements centered around issues related to the transition from a Reynolds-averaged state to one with resolved turbulent content. Improvements to the inflow conditions are suggested as a remedy to this dilemma. Comparisons between resolved second-order turbulence statistics and their modeled Reynolds-averaged counterparts were also performed.

  14. Detection of a variable number of ribosomal DNA loci by fluorescent in situ hybridization in Populus species.

    Science.gov (United States)

    Prado, E A; Faivre-Rampant, P; Schneider, C; Darmency, M A

    1996-10-01

    Fluorescent in situ hybridization (FISH) was applied to related Populus species (2n = 19) in order to detect rDNA loci. An interspecific variability in the number of hybridization sites was revealed using as probe an homologous 25S clone from Populus deltoides. The application of image analysis methods to measure fluorescence intensity of the hybridization signals has enabled us to characterize major and minor loci in the 18S-5.8S-25S rDNA. We identified one pair of such rDNA clusters in Populus alba; two pairs, one major and one minor, in both Populus nigra and P. deltoides; and three pairs in Populus balsamifera, (two major and one minor) and Populus euroamericana (one major and two minor). FISH results are in agreement with those based on RFLP analysis. The pBG13 probe containing 5S sequence from flax detected two separate clusters corresponding to the two size classes of units that coexist within 5S rDNA of most Populus species. Key words : Populus spp., fluorescent in situ hybridization, FISH, rDNA variability, image analysis.

  15. Biotechnology applied to fish reproduction: tools for conservation.

    Science.gov (United States)

    de Siqueira-Silva, Diógenes Henrique; Saito, Taiju; Dos Santos-Silva, Amanda Pereira; da Silva Costa, Raphael; Psenicka, Martin; Yasui, George Shigueki

    2018-04-29

    This review discusses the new biotechnological tools that are arising and promising for conservation and enhancement of fish production, mainly regarding the endangered and the most economically important species. Two main techniques, in particular, are available to avoid extinction of endangered fish species and to improve the production of commercial species. Germ cell transplantation technology includes a number of approaches that have been studied, such as the transplantation of embryo-to-embryo blastomere, embryo-to-embryo differentiated PGC, larvae to larvae and embryo differentiated PGC, transplantation of spermatogonia from adult to larvae or between adults, and oogonia transplantation. However, the success of germ cell transplantation relies on the prior sterilization of fish, which can be performed at different stages of fish species development by means of several protocols that have been tested in order to achieve the best approach to produce a sterile fish. Among them, fish hybridization and triploidization, germline gene knockdown, hyperthermia, and chemical treatment deserve attention based on important results achieved thus far. This review currently used technologies and knowledge about surrogate technology and fish sterilization, discussing the stronger and the weaker points of each approach.

  16. Annual trends in catchability and fish stock assessments

    DEFF Research Database (Denmark)

    Marchal, P.; Ulrich, Clara; Korsbrekke, K.

    2003-01-01

    . The performances of the new and traditional XSA assessments are compared using criteria based on the precision of catchability estimates, stationarity of Log-catchability residuals and retrospective patterns relative to fishing mortality, spawning stock biomass and recruitment estimates. The performances....... A range of catchability trends, including values derived from the "Hybrid" method, is then implemented to standardise the fishing effort of some tuning fleets used in the stock assessments performed by XSA (eXtended Survivors Analysis). Stocks being assessed are the North Sea cod, saithe, plaice and sole...

  17. Fast-FISH Detection and Semi-Automated Image Analysis of Numerical Chromosome Aberrations in Hematological Malignancies

    Directory of Open Access Journals (Sweden)

    Arif Esa

    1998-01-01

    Full Text Available A new fluorescence in situ hybridization (FISH technique called Fast-FISH in combination with semi-automated image analysis was applied to detect numerical aberrations of chromosomes 8 and 12 in interphase nuclei of peripheral blood lymphocytes and bone marrow cells from patients with acute myelogenous leukemia (AML and chronic lymphocytic leukemia (CLL. Commercially available α-satellite DNA probes specific for the centromere regions of chromosome 8 and chromosome 12, respectively, were used. After application of the Fast-FISH protocol, the microscopic images of the fluorescence-labelled cell nuclei were recorded by the true color CCD camera Kappa CF 15 MC and evaluated quantitatively by computer analysis on a PC. These results were compared to results obtained from the same type of specimens using the same analysis system but with a standard FISH protocol. In addition, automated spot counting after both FISH techniques was compared to visual spot counting after standard FISH. A total number of about 3,000 cell nuclei was evaluated. For quantitative brightness parameters, a good correlation between standard FISH labelling and Fast-FISH was found. Automated spot counting after Fast-FISH coincided within a few percent to automated and visual spot counting after standard FISH. The examples shown indicate the reliability and reproducibility of Fast-FISH and its potential for automatized interphase cell diagnostics of numerical chromosome aberrations. Since the Fast-FISH technique requires a hybridization time as low as 1/20 of established standard FISH techniques, omitting most of the time consuming working steps in the protocol, it may contribute considerably to clinical diagnostics. This may especially be interesting in cases where an accurate result is required within a few hours.

  18. Anisakid nematodes as possible markers to trace fish products

    Directory of Open Access Journals (Sweden)

    Vincenzo Ferrantelli

    2015-03-01

    Full Text Available In this work a total of 949 fish samples were analysed for the identification of nematode larvae belonging to the Anisakidae family. Biomolecular application for the identification of Anisakidae larvae can be an optimal instrument for the traceability of fish products, described on the Reg. EC 178/2002. Results confirm a correlation between geographical distribution of fishes and presence of specific Anisakid larvae. FAO 37 zone (Mediterranean sea showed a prevailing distribution of Anisakis pegreffii and a minimal presence of A. simplex s.s. in hybrid form with Anisakis pegreffii. FAO 27 zone showed a prevailing distribution of A. simplex s.s. in fish like Brosme (Brosme brosme and infestation prevalence of Pseudoterranova krabbei and P. decipiens s.s. in Gadus morhua. Obtained results validate the hypothesis that molecular biology methods for identifying Anisakidae larvae are effective traceability markers of fish products.

  19. Extensive polymorphism and chromosomal characteristics of ribosomal DNA in the characid fish Triportheus venezuelensis (Characiformes, Characidae

    Directory of Open Access Journals (Sweden)

    Mauro Nirchio

    2007-01-01

    Full Text Available The karyotype and chromosomal characteristics of the characid fish Triportheus venezuelensis were investigated using differential staining techniques (C-banding, Ag-NOR staining and fluorescent in situ hybridization (FISH with an 18S rDNA probe. The diploid chromosome number (2n = 52, karyotype composition and sex chromosome determination system of the ZZ/ZW type were the same as previously described in other species of the genus Triportheus. However, extensive variation regarding nucleolus organizer regions (NOR different from other species was observed. 18S rDNA sequences were distributed on nine chromosome pairs, but the number of chromosomes with Ag-NORs was usually lower, reaching a maximum of four chromosomes. When sequential staining experiments were performed, it was demonstrated that: 1. active NORs usually corresponded to segments with 18S rDNA genes identified in FISH experiments; 2. several 18S rDNA sequences were not silver-stained, suggesting that they do not correspond to active NORs; and 3. some chromosomes with silver-stained regions did not display any 18S rDNA signals. These findings characterize an extensive polymorphism associated with the NOR-bearing chromosomes of T. venezuelensis and emphasize the importance of combining traditional and molecular techniques in chromosome studies.

  20. 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.

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

    International Nuclear Information System (INIS)

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

    1995-01-01

    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. Widespread hybridization and bidirectional introgression in sympatric species of coral reef fish

    KAUST Repository

    Harrison, Hugo B.; Berumen, Michael L.; Saenz-Agudelo, Pablo; Salas, Eva; Williamson, David H.; Jones, Geoffrey P.

    2017-01-01

    interspecific hybrids from a collection of 2,991 coral trout sampled in inshore and mid-shelf reefs of the southern Great Barrier Reef. Hybrids were ubiquitous among reefs, fertile and spanned multiple generations suggesting both ecological and evolutionary

  3. Effect of anesthetic, tag size, and surgeon experience on postsurgical recovering after implantation of electronic tags in a neotropical fish: Prochilodus lineatus (Valenciennes, 1837 (Characiformes: Prochilodontidae

    Directory of Open Access Journals (Sweden)

    João M. Lopes

    Full Text Available ABSTRACT Implantation of telemetry transmitters in fish can be affected by different parameters. This study aimed to evaluate the effect of type of anesthetic, tag size, and surgeon experience on surgical and postsurgical wound healing in the neotropical fish Prochilodus lineatus . In total, eighty fish were surgically implanted with telemetry transmitters and forty fish were kept as controls. Forty fish were implanted with a small tag and other forty were implanted with a large tag. Similarly, forty fish were anesthetized with eugenol and forty fish were anesthetized by electroanesthesia, and forty surgeries were performed by an expert surgeon and forty surgeries were performed by novice surgeons. At the end of the experimental period seventeen (21.3% tagged fish had postsurgical complications, including death (1.3%, tag expulsion (2.5%, antenna migration (2.5%, and infection (15%. Tag size was the key determinant for postsurgical complications. Surgical details and postsurgical wound healing were not affected by type of anesthetic. Incision size, duration of surgery, and wound area were significantly affected by tag size and surgeon experience, and the number of sutures was significantly affected by tag size only. The results indicate that successful implantation of telemetry transmitters is dependent upon surgeon experience and tag size.

  4. Overview, Progress, and Plans for the Compact Toroidal Hybrid Experiment

    Science.gov (United States)

    Hartwell, G. J.; Allen, N. R.; Ennis, D. A.; Hanson, J. D.; Howell, E. C.; Johnson, C. A.; Knowlton, S. F.; Kring, J. D.; Ma, X.; Maurer, D. A.; Ross, K. G.; Schmitt, J. C.; Traverso, P. J.; Williamson, E. N.

    2017-10-01

    The Compact Toroidal Hybrid (CTH) is an l = 2 , m = 5 torsatron/tokamak hybrid (R0 = 0.75 m, ap 0.2 m, and | B | disruption studies. The main goals of the CTH experiment are to study disruptive behavior as a function of applied 3D magnetic shaping, and to test and advance the V3FIT reconstruction code and NIMROD modeling of CTH. The disruptive density limit is observed to exceed the Greenwald limit as the vacuum transform is increased with no observed threshold for avoidance. Low-q operations (1.1 routine, with disruptions ceasing if the vacuum transform is raised above 0.07. Sawteeth are observed in CTH and have a similar phenomenology to tokamak sawteeth despite employing a 3D confining field. Application of vacuum transform has been demonstrated to reduce and eliminate the vertical drift of elongated discharges. Internal SXR diagnostics, in conjunction with external magnetics, extend the range of reconstruction accuracy into the plasma core. This work is supported by U.S. Department of Energy Grant No. DE-FG02-00ER54610.

  5. A novel artificial fish swarm algorithm for recalibration of fiber optic gyroscope error parameters.

    Science.gov (United States)

    Gao, Yanbin; Guan, Lianwu; Wang, Tingjun; Sun, Yunlong

    2015-05-05

    The artificial fish swarm algorithm (AFSA) is one of the state-of-the-art swarm intelligent techniques, which is widely utilized for optimization purposes. Fiber optic gyroscope (FOG) error parameters such as scale factors, biases and misalignment errors are relatively unstable, especially with the environmental disturbances and the aging of fiber coils. These uncalibrated error parameters are the main reasons that the precision of FOG-based strapdown inertial navigation system (SINS) degraded. This research is mainly on the application of a novel artificial fish swarm algorithm (NAFSA) on FOG error coefficients recalibration/identification. First, the NAFSA avoided the demerits (e.g., lack of using artificial fishes' pervious experiences, lack of existing balance between exploration and exploitation, and high computational cost) of the standard AFSA during the optimization process. To solve these weak points, functional behaviors and the overall procedures of AFSA have been improved with some parameters eliminated and several supplementary parameters added. Second, a hybrid FOG error coefficients recalibration algorithm has been proposed based on NAFSA and Monte Carlo simulation (MCS) approaches. This combination leads to maximum utilization of the involved approaches for FOG error coefficients recalibration. After that, the NAFSA is verified with simulation and experiments and its priorities are compared with that of the conventional calibration method and optimal AFSA. Results demonstrate high efficiency of the NAFSA on FOG error coefficients recalibration.

  6. Experimental observation of G banding verifying X-ray workers' chromosome translocation detected by FISH

    International Nuclear Information System (INIS)

    Sun Yuanming; Li Jin; Wang Qin; Tang Weisheng; Wang Zhiquan

    2002-01-01

    Objective: FISH is the most effective way of detecting chromosome aberration and many factors affect its accuracy. G-banding is used to verify the results of early X-ray workers' chromosome translocation examined by FISH. Methods: The chromosome translocations of early X-ray workers have been analysed by FISH (fluorescence in situ hybridization) and G-banding, yields of translocation treated with statistics. Results: The chromosome aberrations frequencies by tow methods are closely related. Conclusion: FISH is a feasible way to analyse chromosome aberrations of X-ray workers and reconstruct dose

  7. Recent progress in lower hybrid current drive theory and experiments

    International Nuclear Information System (INIS)

    Barbato, E.

    1998-01-01

    In this paper lower hybrid current drive (LHCD) experimental milestones paving the way for future experiments are briefly summarized. The current drive efficiency scaling with the electron temperature is discussed. The role of wave propagation in determining the power deposition profile is stressed and methods are discussed to control the current density profile. Modelling of negative central shear configurations, experimentally obtained by LHCD, are reported. A good agreement is found between the modelling results and the experimental findings, thus showing that a good degree of understanding has been achieved in LHCD theory. (author)

  8. Retention of habitat complexity minimizes disassembly of reef fish communities following disturbance: a large-scale natural experiment.

    Directory of Open Access Journals (Sweden)

    Michael J Emslie

    Full Text Available High biodiversity ecosystems are commonly associated with complex habitats. Coral reefs are highly diverse ecosystems, but are under increasing pressure from numerous stressors, many of which reduce live coral cover and habitat complexity with concomitant effects on other organisms such as reef fishes. While previous studies have highlighted the importance of habitat complexity in structuring reef fish communities, they employed gradient or meta-analyses which lacked a controlled experimental design over broad spatial scales to explicitly separate the influence of live coral cover from overall habitat complexity. Here a natural experiment using a long term (20 year, spatially extensive (∼ 115,000 kms(2 dataset from the Great Barrier Reef revealed the fundamental importance of overall habitat complexity for reef fishes. Reductions of both live coral cover and habitat complexity had substantial impacts on fish communities compared to relatively minor impacts after major reductions in coral cover but not habitat complexity. Where habitat complexity was substantially reduced, species abundances broadly declined and a far greater number of fish species were locally extirpated, including economically important fishes. This resulted in decreased species richness and a loss of diversity within functional groups. Our results suggest that the retention of habitat complexity following disturbances can ameliorate the impacts of coral declines on reef fishes, so preserving their capacity to perform important functional roles essential to reef resilience. These results add to a growing body of evidence about the importance of habitat complexity for reef fishes, and represent the first large-scale examination of this question on the Great Barrier Reef.

  9. Manufacturing experience and test results of the PS prototype flexible hybrid circuit for the CMS Tracker Upgrade

    CERN Document Server

    Kovacs, Mark Istvan; Gadek, Tomasz; Honma, Alan; Vasey, Francois

    2017-01-01

    The CMS Tracker Phase-2 Upgrade for HL-LHC requires High Density Interconnect (HDI) flexible hybrid circuits to build modules with low mass and high granularity. The hybrids are carbon fibre reinforced flexible circuits with flip-chips and passives. Three different manufacturers produced prototype hybrids for the Pixel-Strip type modules. The first part of the presentation will focus on the design challenges of this state of the art circuit. Afterwards, the difficulties and experience related to the circuit manufacturing and assembly are presented. The description of quality inspection methods with comprehensive test results will lead to the conclusion.

  10. Continental-scale patterns of nutrient and fish effects on shallow lakes: synthesis of a pan-European mesocosm experiment

    NARCIS (Netherlands)

    Moss, B.; Stephen, D.; Balayla, D.; Bécares, E.; Collings, S.E.; Fernández-Aláez, C.; Fernández-Aláez, M.; Ferriol, C.; García, P.; Gomá, J.; Gyllström, M.; Hansson, L-A.; Hietala, J.; Kairesalo, T.; Rosa Miracle, M.; Romo, S.; Rueda, J.; Russell, V.; Ståhl-Delbanco, A.; Svensson, M.; Vakkilainen, K.; Valentín, M.; van de Bund, W.; Van Donk, E.; Vicente, E.; Villena, M.J.

    2004-01-01

    1. Results are analysed from 11 experiments in which effects of fish addition and nutrient loading on shallow lakes were studied in mesocosms. The experiments, five in 1998, six in 1999, were carried out in six lakes, distributed from Finland to southern Spain, according to a standard protocol. 2.

  11. Evaluation of hybrid neutralization/biosorption process for zinc ions removal from automotive battery effluent by dolomite and fish scales.

    Science.gov (United States)

    Ribeiro, C; Scheufele, F B; Alves, H J; Kroumov, A D; Espinoza-Quiñones, F R; Módenes, A N; Borba, C E

    2018-02-26

    This work focused in the evaluation of Oreochromis niloticus fish scales (FS) as biosorbent material in the removal of Zn from a synthetic effluent based on automotive battery industry effluent and, further, a hybrid neutralization/biosorption process, aiming at a high-quality treated effluent, by a cooperative use of dolomite and FS. For this, a physicochemical and morphological characterization (i.e. SEM-EDX, FTIR, XRD, and TXRF) was performed, which helped to clarify a great heterogeneity of active sites (phosphate, carbonate, amide, and hydroxyl) on the biosorbent; also the inorganic constituents (apatites) leaching from the FS was identified. Biosorption results pointed out to a pH-dependent process due to changes in the functional group's anionic character (i.e. electrostatic interactions), where an initial pH = 3 favored the Zn uptake. Kinetic and equilibrium studies confirmed the heterogeneous surface and cooperative sorption, wherein experimental data were described by Generalized Elovich kinetic model and the favorable isotherm profile by Langmuir-Freundlich isotherm ([Formula: see text] = 15.38 mg g -1 and [Formula: see text]). Speciation diagram of Zn species along with the leached species demonstrated that, for the studied pH range, the biosorption was the most likely phenomena rather than precipitation. Finally, the hybrid neutralization/biosorption process showed great potential since both the Zn concentration levels and the pH reached the legislation standards (C Zn  = 4 mg L -1 ; pH = 5). Hence, based on the characterization and biosorption results, a comprehensive evaluation of the involved mechanisms in such complex system helped to verify the prospective of FS biosorbent for the Zn treatment from solution, in both individual and hybrid processes.

  12. Exploring the origin of the D genome of oat by fluorescence in situ hybridization.

    Science.gov (United States)

    Luo, Xiaomei; Zhang, Haiqin; Kang, Houyang; Fan, Xing; Wang, Yi; Sha, Lina; Zhou, Yonghong

    2014-09-01

    Further understanding of the origin of cultivated oat would accelerate its genetic improvement. In particular, it would be useful to clarify which diploid progenitor contributed the D genome of this allohexaploid species. In this study, we demonstrate that the landmarks produced by fluorescence in situ hybridization (FISH) of species of Avena using probes derived from Avena sativa can be used to explore the origin of the D genome. Selected sets of probes were hybridized in several sequential experiments performed on exactly the same chromosome spreads, with multiple probes of cytological preparations. Probes pITS and A3-19 showed there might be a similar distribution of pITS between the Ac and D genomes. These results indicated that the Ac genome is closely related to the D genome, and that Avena canariensis (AcAc) could be the D-genome donor of cultivated oat.

  13. Prevalence of fish diseases in fish farms in Katsina State, Nigeria ...

    African Journals Online (AJOL)

    Prevalence of fish disease in fish farms in Nigeria was investigated using data from structured questionnaire obtained during farm visits, and analysed using descriptive statistics and multiple linear regression. Findings from the study revealed that majority of the farm manager's (45.7) have less than 3 years of experience in ...

  14. Integrating a FISH imaging system into the cytology laboratory

    Directory of Open Access Journals (Sweden)

    Denice Smith G

    2010-01-01

    Full Text Available We have implemented an interactive imaging system for the interpretation of UroVysion fluorescence in situ hybridization (FISH to improve throughput, productivity, quality control and diagnostic accuracy. We describe the Duet imaging system, our experiences with implementation, and outline the financial investment, space requirements, information technology needs, validation, and training of cytotechnologists needed to integrate such a system into a cytology laboratory. Before purchasing the imaging system, we evaluated and validated the instrument at our facility. Implementation required slide preparation changes, IT modifications, development of training programs, and revision of job descriptions for cytotechnologists. A darkened room was built to house the automated scanning station and microscope, as well as two imaging stations. IT changes included generation of storage for archival images on the LAN, addition of external hard drives for back-up, and changes to cable connections for communication between remote locations. Training programs for cytotechnologists, and pathologists/fellows/residents were developed, and cytotechnologists were integrated into multiple steps of the process. The imaging system has resulted in increased productivity for pathologists, concomitant with an expanded role of cytotechnologists in multiple critical steps, including FISH, scan setup, reclassification, and initial interpretation.

  15. Interaction of fish aryl hydrocarbon receptor paralogs (AHR1 and AHR2) with the retinoblastoma protein

    Energy Technology Data Exchange (ETDEWEB)

    Merson, Rebeka R., E-mail: rmerson@ric.edu [Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543 (United States); Biology Department, Rhode Island College, 500 Mt. Pleasant Ave., Providence, RI 02908 (United States); Karchner, Sibel I.; Hahn, Mark E. [Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543 (United States)

    2009-08-13

    The aryl hydrocarbon receptor (AHR) mediates the toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds. In some mammalian cell lines, TCDD induces G1 cell cycle arrest, which depends on an interaction between the AHR and the retinoblastoma tumor suppressor (RB). Mammals possess one AHR, whereas fishes possess two or more AHR paralogs that differ in the domains important for AHR-RB interactions in mammals. To test the hypothesis that fish AHR paralogs differ in their ability to interact with RB, we cloned RB cDNA from Atlantic killifish, Fundulus heteroclitus, and studied the interactions of killifish RB protein with killifish AHR1 and AHR2. In coimmunoprecipitation experiments, in vitro-expressed killifish RB coprecipitated with both AHR1 and AHR2. Consistent with these results, both killifish AHR1 and AHR2 interacted with RB in mammalian two-hybrid assays. These results suggest that both fish AHR1 and AHR2 paralogs may have the potential to influence cell proliferation through interactions with RB.

  16. Hybridization, agency discretion, and implementation of the U.S. Endangered Species Act.

    Science.gov (United States)

    Lind-Riehl, Jennifer F; Mayer, Audrey L; Wellstead, Adam M; Gailing, Oliver

    2016-12-01

    The U.S. Endangered Species Act (ESA) requires that the "best available scientific and commercial data" be used to protect imperiled species from extinction and preserve biodiversity. However, it does not provide specific guidance on how to apply this mandate. Scientific data can be uncertain and controversial, particularly regarding species delineation and hybridization issues. The U.S. Fish and Wildlife Service (FWS) had an evolving hybrid policy to guide protection decisions for individuals of hybrid origin. Currently, this policy is in limbo because it resulted in several controversial conservation decisions in the past. Biologists from FWS must interpret and apply the best available science to their recommendations and likely use considerable discretion in making recommendations for what species to list, how to define those species, and how to recover them. We used semistructured interviews to collect data on FWS biologists' use of discretion to make recommendations for listed species with hybridization issues. These biologists had a large amount of discretion to determine the best available science and how to interpret it but generally deferred to the scientific consensus on the taxonomic status of an organism. Respondents viewed hybridization primarily as a problem in the context of the ESA, although biologists who had experience with hybridization issues were more likely to describe it in more nuanced terms. Many interviewees expressed a desire to continue the current case-by-case approach for handling hybridization issues, but some wanted more guidance on procedures (i.e., a "flexible" hybrid policy). Field-level information can provide critical insight into which policies are working (or not working) and why. The FWS biologists' we interviewed had a high level of discretion, which greatly influenced ESA implementation, particularly in the context of hybridization. © 2016 Society for Conservation Biology.

  17. Experiments on the use of sound as a fish deterrent

    International Nuclear Information System (INIS)

    Turnpenny, A.W.H.; Thatcher, K.P.; Wood, R.; Loeffelman, P.H.

    1993-01-01

    This report describes a series of experimental studies into the potential use of acoustic stimuli to deter fish from water intakes at thermal and hydroelectric power stations. The aim was to enlarge the range of candidate signals for testing, and to apply these in more rigorous laboratory trials and to a wider range of estuarine and marine fish species than was possible in previous initial preliminary studies. The trials were also required to investigate the degree to which fish might become habituated to the sound signals, consequently reducing their effectiveness. The species of fish which were of interest in this study were the Atlantic salmon (Salmo salar), sea trout (Salmo trutta), the shads (Alosa fallax, A. alosa), the European eel (Anguilla anguilla), bass (Dicentrarchus labrax), herring (Clupea harengus), whiting (Merlangius merlangus) and cod (Gadus morhua). All of these species are considered to be of conservation and/or commercial importance in Britain today and are potentially vulnerable to capture by nuclear, fossil-fuelled and tidal generating stations. Based on the effectiveness of the signals observed in these trials, a properly developed and sited acoustic fish deterrent system is expected to reduce fish impingement significantly at water intakes. Field trials at an estuarine power station are recommended. (author)

  18. Efficiency of fluorescence in situ hybridization for bacterial cell identification in temporary river sediments with contrasting water content.

    Science.gov (United States)

    Fazi, Stefano; Amalfitano, Stefano; Pizzetti, Ilaria; Pernthaler, Jakob

    2007-09-01

    We studied the efficiency of two hybridization techniques for the analysis of benthic bacterial community composition under varying sediment water content. Microcosms were set up with sediments from four European temporary rivers. Wet sediments were dried, and dry sediments were artificially rewetted. The percentage of bacterial cells detected by fluorescence in situ hybridization with fluorescently monolabeled probes (FISH) significantly increased from dry to wet sediments, showing a positive correlation with the community activity measured via incorporation of (3)H leucine. FISH and signal amplification by catalyzed reporter deposition (CARD-FISH) could significantly better detect cells with low activity in dried sediments. Through the application of an optimized cell permeabilization protocol, the percentage of hybridized cells by CARD-FISH showed comparable values in dry and wet conditions. This approach was unrelated to (3)H leucine incorporation rates. Moreover, the optimized protocol allowed a significantly better visualization of Gram-positive Actinobacteria in the studied samples. CARD-FISH is, therefore, proposed as an effective technique to compare bacterial communities residing in sediments with contrasting water content, irrespective of differences in the activity state of target cells. Considering the increasing frequencies of flood and drought cycles in European temporary rivers, our approach may help to better understand the dynamics of microbial communities in such systems.

  19. Clinical consequences of using PNA-FISH in Staphylococcal bacteraemia

    DEFF Research Database (Denmark)

    Laub, R R; Knudsen, Inge Jenny Dahl

    2014-01-01

    To optimize patient treatment and rational use of antimicrobials, it is important to provide fast information on findings in blood-cultures (BCs). The purpose of this study was to evaluate the impact of using peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) on positive BCs conta...

  20. Cytogenetics of Festulolium (Festuca x Lolium hybrids)

    Czech Academy of Sciences Publication Activity Database

    Kopecký, David; Lukaszewski, A.J.; Doležel, Jaroslav

    2008-01-01

    Roč. 120, 3-4 (2008), s. 370-383 ISSN 1424-8581 R&D Projects: GA ČR GP521/07/P479; GA MZe QH71267 Institutional research plan: CEZ:AV0Z50380511 Keywords : Grasses * hybrids * FISH Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.965, year: 2008

  1. Use of Hybridization Chain Reaction-Fluorescent In Situ Hybridization To Track Gene Expression by Both Partners during Initiation of Symbiosis.

    Science.gov (United States)

    Nikolakakis, K; Lehnert, E; McFall-Ngai, M J; Ruby, E G

    2015-07-01

    The establishment of a productive symbiosis between Euprymna scolopes, the Hawaiian bobtail squid, and its luminous bacterial symbiont, Vibrio fischeri, is mediated by transcriptional changes in both partners. A key challenge to unraveling the steps required to successfully initiate this and many other symbiotic associations is characterization of the timing and location of these changes. We report on the adaptation of hybridization chain reaction-fluorescent in situ hybridization (HCR-FISH) to simultaneously probe the spatiotemporal regulation of targeted genes in both E. scolopes and V. fischeri. This method revealed localized, transcriptionally coregulated epithelial cells within the light organ that responded directly to the presence of bacterial cells while, at the same time, provided a sensitive means to directly show regulated gene expression within the symbiont population. Thus, HCR-FISH provides a new approach for characterizing habitat transition in bacteria and for discovering host tissue responses to colonization. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  2. Comparison of Fluorescence In Situ Hybridization and Chromogenic In Situ Hybridization for Low and High Throughput HER2 Genetic Testing

    Science.gov (United States)

    Poulsen, Tim S.; Espersen, Maiken L. M.; Kofoed, Vibeke; Dabetic, Tanja; Høgdall, Estrid; Balslev, Eva

    2013-01-01

    The purpose was to evaluate and compare 5 different HER2 genetic assays with different characteristics that could affect the performance to analyze the human epidermal growth factor 2 (HER2) gene copy number under low and high throughput conditions. The study included 108 tissue samples from breast cancer patients with HER2 immunohistochemistry (IHC) results scored as 0/1+, 2+, and 3+. HER2 genetic status was analysed using chromogenic in situ hybridization (CISH) and fluorescence in situ hybridization (FISH). Scoring results were documented through digital image analysis. The cancer region of interest was identified from a serial H&E stained slide following tissue cores were transferred to a tissue microarrays (TMA). When using TMA in a routine flow, all patients will be tested for HER2 status with IHC followed by CISH or FISH, thereby providing individual HER2 results. In conclusion, our results show that the differences between the HER2 genetic assays do not have an effect on the analytic performance and the CISH technology is superior to high throughput HER2 genetic testing due to scanning speed, while the IQ-FISH may still be a choice for fast low throughput HER2 genetic testing. PMID:24383005

  3. Mesocosm experiments on tritium dynamics in carp fish

    International Nuclear Information System (INIS)

    Reji, T.K.; Vishnu, M.S.; Joshi, R.M.; Dileep, B.N.; Baburajan, A.; Ravi, P.M.

    2013-01-01

    Tritium dynamics in carp fish (Cyprinus carpio) was studied in a locally designed mesocosm simulating a lake condition. The fishes were reared in an experimental tank containing tritiated water. Tissue Free water tritium (TFWT) concentration and Organically Bound Tritium (OBT) was measured for 3 months period. TFWT reached equilibrium with exposed water within one day. Detectable amount of OBT was observed after two months of exposure. OBT to TFWT ratio was 0.1. Estimated OBT was in agreement with that calculated using IAEA specific activity model. (author)

  4. Are hybrids between Atlantic salmon and brown trout suitable long-term hosts of Gyrodactylus salaris during winter?

    Science.gov (United States)

    Knudsen, R; Henriksen, E H; Gjelland, K Ø; Hansen, H; Hendrichsen, D K; Kristoffersen, R; Olstad, K

    2017-10-01

    The monogenean parasite Gyrodactylus salaris poses serious threats to many Atlantic salmon populations and presents many conservation and management questions/foci and challenges. It is therefore critical to identify potential vectors for infection. To test whether hybrids of native Atlantic salmon (Salmo salar) × brown trout (Salmo trutta) are suitable as reservoir hosts for G. salaris during winter, infected hybrid parr were released into a natural subarctic brook in the autumn. Six months later, 23.9% of the pit-tagged fish were recaptured. During the experimental period, the hybrids had a sixfold increase in mean intensity of G. salaris, while the prevalence decreased from 81% to 35%. There was high interindividual hybrid variability in susceptibility to infections. The maximum infrapopulation growth rate (0.018 day -1 ) of G. salaris throughout the winter was comparable to earlier laboratory experiments at similar temperatures. The results confirm that infrapopulations of G. salaris may reproduce on a hybrid population for several generations at low water temperatures (~1 °C). Wild salmon-trout hybrids are undoubtedly susceptible to G. salaris and represent an important reservoir host for the parasite independent of other co-occurring susceptible hosts. Consequently, these hybrids may pose a serious risk for G. salaris transmission to nearby, uninfected rivers by migratory individuals. © 2017 John Wiley & Sons Ltd.

  5. Experiments on radionuclide accumulation by fishes from the Angra Dos Reis region of Brazil

    International Nuclear Information System (INIS)

    Guimaraes, J.R.D.

    1988-11-01

    To follow the bioaccumulation and tissue distribution of 137 Cs and 60 Co in fish from tropical waters near the Angra nuclear power station in Brazil, juvenile grouper (Epinephelus sp.) were exposed under laboratory conditions to Angra sea water containing the two radionuclides initially added in chloride form. During the 4-month exposure period, the sea water was not changed and the fish received regular rations of non-radioactive food. Following uptake, the fish were dissected and composite tissue samples monitored for radioactivity. 137 Cs concentration factors (CFs) were 3 and 5 in whole fish and muscle, respectively. The highest concentration of 137 Cs was found in the muscle which accounted for 66% of the total body burden. In contrast, the highest concentration and greatest fraction (19%) of 60 Co was located in the liver. Whole body and muscle CFs for 60 Co, 0.2 and 0.08 respectively, were much lower than those for 137 Cs. Compared with published data, only very small amounts of both radionuclides were taken up by bone in the present study. This observation and the fact that much higher 137 Cs CFs have been reported in fish from field studies suggests that uptake through the food chain is the principal pathway for bioaccumulation in nature. Elution of freshly-prepared and previously-used radioactive sea water through chelating resins indicated that 60 Co added in the cationic form was almost totally converted to neutral or anionic forms after four months. Additional experiments carried out with the previously-used experimental media demonstrated that the possibly ligand-bound 60 Co was approximately 2.5 times more available for uptake by grouper than the cationic forms initially present. It is concluded that the uptake of 60 Co by fish exposed to radioactive effluents would be a function of the different forms of 60 Co present. 20 refs, 2 figs, 2 tabs

  6. Sugar Cane Genome Numbers Assumption by Ribosomal DNA FISH Techniques

    NARCIS (Netherlands)

    Thumjamras, S.; Jong, de H.; Iamtham, S.; Prammanee, S.

    2013-01-01

    Conventional cytological method is limited for polyploidy plant genome study, especially sugar cane chromosomes that show unstable numbers of each cultivar. Molecular cytogenetic as fluorescent in situ hybridization (FISH) techniques were used in this study. A basic chromosome number of sugar cane

  7. San Juanico Hybrid System Technical and Institutional Assessment: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Corbus, D.; Newcomb, C.; Yewdall, Z.

    2004-07-01

    San Juanico is a fishing village of approximately 120 homes in the Municipality of Comondu, Baja California. In April, 1999, a hybrid power system was installed in San Juanico to provide 24-hour power, which was not previously available. Before the installation of the hybrid power system, a field study was conducted to characterize the electrical usage and institutional and social framework of San Juanico. One year after the installation of the hybrid power system a''post-electrification'' study was performed to document the changes that had occurred after the installation. In December of 2003, NREL visited the site to conduct a technical assessment of the system.

  8. Towards Fluorescence In Vivo Hybridization (FIVH) Detection of H. pylori in Gastric Mucosa Using Advanced LNA Probes

    Science.gov (United States)

    Fontenete, Sílvia; Leite, Marina; Guimarães, Nuno; Madureira, Pedro; Ferreira, Rui Manuel; Figueiredo, Céu; Wengel, Jesper; Azevedo, Nuno Filipe

    2015-01-01

    In recent years, there have been several attempts to improve the diagnosis of infection caused by Helicobacter pylori. Fluorescence in situ hybridization (FISH) is a commonly used technique to detect H. pylori infection but it requires biopsies from the stomach. Thus, the development of an in vivo FISH-based method (FIVH) that directly detects and allows the visualization of the bacterium within the human body would significantly reduce the time of analysis, allowing the diagnosis to be performed during endoscopy. In a previous study we designed and synthesized a phosphorothioate locked nucleic acid (LNA)/ 2’ O-methyl RNA (2’OMe) probe using standard phosphoramidite chemistry and FISH hybridization was then successfully performed both on adhered and suspended bacteria at 37°C. In this work we simplified, shortened and adapted FISH to work at gastric pH values, meaning that the hybridization step now takes only 30 minutes and, in addition to the buffer, uses only urea and probe at non-toxic concentrations. Importantly, the sensitivity and specificity of the FISH method was maintained in the range of conditions tested, even at low stringency conditions (e.g., low pH). In conclusion, this methodology is a promising approach that might be used in vivo in the future in combination with a confocal laser endomicroscope for H. pylori visualization. PMID:25915865

  9. Towards Fluorescence In Vivo Hybridization (FIVH Detection of H. pylori in Gastric Mucosa Using Advanced LNA Probes.

    Directory of Open Access Journals (Sweden)

    Sílvia Fontenete

    Full Text Available In recent years, there have been several attempts to improve the diagnosis of infection caused by Helicobacter pylori. Fluorescence in situ hybridization (FISH is a commonly used technique to detect H. pylori infection but it requires biopsies from the stomach. Thus, the development of an in vivo FISH-based method (FIVH that directly detects and allows the visualization of the bacterium within the human body would significantly reduce the time of analysis, allowing the diagnosis to be performed during endoscopy. In a previous study we designed and synthesized a phosphorothioate locked nucleic acid (LNA/ 2' O-methyl RNA (2'OMe probe using standard phosphoramidite chemistry and FISH hybridization was then successfully performed both on adhered and suspended bacteria at 37°C. In this work we simplified, shortened and adapted FISH to work at gastric pH values, meaning that the hybridization step now takes only 30 minutes and, in addition to the buffer, uses only urea and probe at non-toxic concentrations. Importantly, the sensitivity and specificity of the FISH method was maintained in the range of conditions tested, even at low stringency conditions (e.g., low pH. In conclusion, this methodology is a promising approach that might be used in vivo in the future in combination with a confocal laser endomicroscope for H. pylori visualization.

  10. Evaluation of Myc Gene Amplification in Prostate Cancer Using a Dual Color Chromogenic in-Situ Hybridization (Dual CISH) Assay

    OpenAIRE

    Daniel Lerda; Marta Cabrera; Jorge Flores; Luis Gutierrez; Armando Chierichetti; Martin Revol; Hernan Garcia Onto

    2013-01-01

    Objetive: The overall purpose of the study was to demonstrate applicability of the Dako dual-color chromogenic in situ hybridization (CISH) assay (DAKO Denmark, Glostrup) with respect to fluorescence in situ hybridization (FISH) probes MYC-C. Methods: MYC gene amplification by FISH and Dako dual-color CISH Results: The study showed that the dual-color CISH assay can convert Texas red and fluorescein isothiocyanate (FITC) signals into chromogenic signals. The dual –color CISH assay was p...

  11. Responses of phytoplankton to fish predation and nutrient loading in shallow lakes: a pan-European mesocosm experiment

    NARCIS (Netherlands)

    van de Bund, W.; Romo, S.; Villena, M.J.; Valentín, M.; Van Donk, E.; Vicente, E.; Vakkilainen, K.; Svensson, M.; Stephen, D.; Ståhl-Delbanco, A.; Rueda, J.; Moss, B.; Rosa Miracle, M.; Kairesalo, T.; Hansson, L-A.; Hietala, J.; Gyllström, M.; Goma, J.; García, P.; Fernández-Aláez, M.; Fernández-Aláez, C.; Ferriol, C.; Collings, S.E.; Bécares, E.; Balayla, D.; Alfonso, T.

    2004-01-01

    1. The impacts of nutrients (phosphorus and nitrogen) and planktivorous fish on phytoplankton composition and biomass were studied in six shallow, macrophyte-dominated lakes across Europe using mesocosm experiments. 2. Phytoplankton biomass was more influenced by nutrients than by densities of

  12. Immunoglobulin heavy-chain fluorescence in situ hybridization-chromogenic in situ hybridization DNA probe split signal in the clonality assessment of lymphoproliferative processes on cytological samples.

    Science.gov (United States)

    Zeppa, Pio; Sosa Fernandez, Laura Virginia; Cozzolino, Immacolata; Ronga, Valentina; Genesio, Rita; Salatiello, Maria; Picardi, Marco; Malapelle, Umberto; Troncone, Giancarlo; Vigliar, Elena

    2012-12-25

    The human immunoglobulin heavy-chain (IGH) locus at chromosome 14q32 is frequently involved in different translocations of non-Hodgkin lymphoma (NHL), and the detection of any breakage involving the IGH locus should identify a B-cell NHL. The split-signal IGH fluorescence in situ hybridization-chromogenic in situ hybridization (FISH-CISH) DNA probe is a mixture of 2 fluorochrome-labeled DNAs: a green one that binds the telomeric segment and a red one that binds the centromeric segment, both on the IGH breakpoint. In the current study, the authors tested the capability of the IGH FISH-CISH DNA probe to detect IGH translocations and diagnose B-cell lymphoproliferative processes on cytological samples. Fifty cytological specimens from cases of lymphoproliferative processes were tested using the split-signal IGH FISH-CISH DNA probe and the results were compared with light-chain assessment by flow cytometry (FC), IGH status was tested by polymerase chain reaction (PCR), and clinicohistological data. The signal score produced comparable results on FISH and CISH analysis and detected 29 positive, 15 negative, and 6 inadequate cases; there were 29 true-positive cases (66%), 9 true-negative cases (20%), 6 false-negative cases (14%), and no false-positive cases (0%). Comparing the sensitivity of the IGH FISH-CISH DNA split probe with FC and PCR, the highest sensitivity was obtained by FC, followed by FISH-CISH and PCR. The split-signal IGH FISH-CISH DNA probe is effective in detecting any translocation involving the IGH locus. This probe can be used on different samples from different B-cell lymphoproliferative processes, although it is not useful for classifying specific entities. Cancer (Cancer Cytopathol) 2012;. © 2012 American Cancer Society. Copyright © 2012 American Cancer Society.

  13. A hybrid ARIMA and neural network model applied to forecast catch volumes of Selar crumenophthalmus

    Science.gov (United States)

    Aquino, Ronald L.; Alcantara, Nialle Loui Mar T.; Addawe, Rizavel C.

    2017-11-01

    The Selar crumenophthalmus with the English name big-eyed scad fish, locally known as matang-baka, is one of the fishes commonly caught along the waters of La Union, Philippines. The study deals with the forecasting of catch volumes of big-eyed scad fish for commercial consumption. The data used are quarterly caught volumes of big-eyed scad fish from 2002 to first quarter of 2017. This actual data is available from the open stat database published by the Philippine Statistics Authority (PSA)whose task is to collect, compiles, analyzes and publish information concerning different aspects of the Philippine setting. Autoregressive Integrated Moving Average (ARIMA) models, Artificial Neural Network (ANN) model and the Hybrid model consisting of ARIMA and ANN were developed to forecast catch volumes of big-eyed scad fish. Statistical errors such as Mean Absolute Errors (MAE) and Root Mean Square Errors (RMSE) were computed and compared to choose the most suitable model for forecasting the catch volume for the next few quarters. A comparison of the results of each model and corresponding statistical errors reveals that the hybrid model, ARIMA-ANN (2,1,2)(6:3:1), is the most suitable model to forecast the catch volumes of the big-eyed scad fish for the next few quarters.

  14. Automation of ALK gene rearrangement testing with fluorescence in situ hybridization (FISH): a feasibility study.

    Science.gov (United States)

    Zwaenepoel, Karen; Merkle, Dennis; Cabillic, Florian; Berg, Erica; Belaud-Rotureau, Marc-Antoine; Grazioli, Vittorio; Herelle, Olga; Hummel, Michael; Le Calve, Michele; Lenze, Dido; Mende, Stefanie; Pauwels, Patrick; Quilichini, Benoit; Repetti, Elena

    2015-02-01

    In the past several years we have observed a significant increase in our understanding of molecular mechanisms that drive lung cancer. Specifically in the non-small cell lung cancer sub-types, ALK gene rearrangements represent a sub-group of tumors that are targetable by the tyrosine kinase inhibitor Crizotinib, resulting in significant reductions in tumor burden. Phase II and III clinical trials were performed using an ALK break-apart FISH probe kit, making FISH the gold standard for identifying ALK rearrangements in patients. FISH is often considered a labor and cost intensive molecular technique, and in this study we aimed to demonstrate feasibility for automation of ALK FISH testing, to improve laboratory workflow and ease of testing. This involved automation of the pre-treatment steps of the ALK assay using various protocols on the VP 2000 instrument, and facilitating automated scanning of the fluorescent FISH specimens for simplified enumeration on various backend scanning and analysis systems. The results indicated that ALK FISH can be automated. Significantly, both the Ikoniscope and BioView system of automated FISH scanning and analysis systems provided a robust analysis algorithm to define ALK rearrangements. In addition, the BioView system facilitated consultation of difficult cases via the internet. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. EGFR status in oral squamous cell carcinoma: comparing immunohistochemistry, FISH and CISH detection in a case series study.

    Science.gov (United States)

    Bernardes, Vanessa Fátima; Gleber-Netto, Frederico Omar; Sousa, Sílvia Ferreira de; Rocha, Rafael Malagoli; Aguiar, Maria Cássia Ferreira de

    2013-01-28

    To compare the immunohistochemistry (IHC) expression of epidermal growth factor receptor (EGFR) in oral squamous cell carcinomas (OSCC) with the gene amplification evaluated by fluorescence in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) and their association with clinicopathological parameters. Additionally, we tested the sensibility and specificity of CISH in comparison with FISH. Case series study Oral surgery and pathology department in a school of dentistry. 52 patients with histopathological diagnosis of OSCC. Tumour tissue samples from 52 patients with OSCC were evaluated by IHC, FISH and CISH using tissue microarray technology. Clinicopathological data from all patients were collected. EGFR+ rates were 53.8% (28/52) by IHC, 5.8% (3/52) by CISH and 15.4% (8/52) by FISH. Amplification detected by CISH and FISH with IHC negative occurred in 3.8% (2/52), and one case (1.9%) showed amplification detected by CISH and FISH and protein overexpression concomitantly. There were 9.6% FISH+ cases with IHC and CISH negative rates and 6/8 (75%) FISH+ and also EGFR+ cases; however, an association between protein expression and gene amplification was not found for both techniques. IHC and FISH rates were not associated with clinicopathological features. CISH+ rates were associated with T3-T4 status. Compared with FISH assay, CISH reached a sensitivity of 37.5% and specificity of 100%. There is no association between EGFR expression and gene amplification in OSCC when the IHC is driven to external epitopes of the protein. Although CISH demonstrates specificity, technical problems may influence sensibility when compared with FISH.

  16. Fish allergens at a glance: Variable allergenicity of parvalbumins, the major fish allergens

    OpenAIRE

    Annette eKuehn; Ines eSwoboda; Karthik eArumugam; Christiane eHilger; François eHentges; François eHentges

    2014-01-01

    Fish is a common trigger of severe, food-allergic reactions. Only a limited number of proteins induce specific IgE-mediated immune reactions. The major fish allergens are the parvalbumins. They are members of the calcium-binding EF-hand protein family characterized by a conserved protein structure. They represent highly cross-reactive allergens for patients with specific IgE to conserved epitopes. These patients might experience clinical reactions with various fish species. On the other hand,...

  17. Fish Allergens at a Glance: Variable Allergenicity of Parvalbumins, the Major Fish Allergens

    OpenAIRE

    Kuehn, Annette; Swoboda, Ines; Arumugam, Karthik; Hilger, Christiane; Hentges, François

    2014-01-01

    Fish is a common trigger of severe, food-allergic reactions. Only a limited number of proteins induce specific IgE-mediated immune reactions. The major fish allergens are the parvalbumins. They are members of the calcium-binding EF-hand protein family characterized by a conserved protein structure. They represent highly cross-reactive allergens for patients with specific IgE to conserved epitopes. These patients might experience clinical reactions with various fish species. On the other hand,...

  18. Fluorescence in situ hybridization (FISH screening for the 22q11.2 deletion in patients with clinical features of velocardiofacial syndrome but without cardiac anomalies

    Directory of Open Access Journals (Sweden)

    Paula Sandrin-Garcia

    2007-01-01

    Full Text Available The velocardiofacial syndrome (VCFS, a condition associated with 22q11.2 deletions, is characterized by a typical facies, palatal anomalies, learning disabilities, behavioral disturbances and cardiac defects. We investigated the frequency of these chromosomal deletions in 16 individuals with VCFS features who presented no cardiac anomalies, one of the main characteristics of VCFS. Fluorescent in situ hybridization (FISH with the N25 (D22S75; 22q11.2 probe revealed deletions in ten individuals (62%. Therefore, even in the absence of cardiac anomalies testing for the 22q11.2 microdeletions in individuals showing other clinical features of this syndrome is recommended.

  19. Identification of bacteria used for microbial enhanced oil recovery process by fluorescence in situ hybridization technique

    Energy Technology Data Exchange (ETDEWEB)

    Fujiwara, K.; Tanaka, S.; Otsuka, M. [Kansai Research Institute, Kyoto (Japan). Lifescience Lab.; Yonebayashi, H. [Japan National Oil Corp., Chiba (Japan). Tech. Research Center; Enomoto, H. [Tohoku University, Sendai (Japan). Dept. of Geoscience and Tech.

    2000-01-01

    A fluorescence in situ hybridization (FISH) technique using 16S rRNA-targeted oligonucleotide probes was developed for rapid detection of microorganisms for use in the microbial enhancement of oil recovery (MEOR) process. Two microorganisms, Enterobacter cloacae TRC-322 and Bacillus licheniformis TRC-18-2-a, were selected from a collection of Enterobacter sp. and Bacillus sp. which were screened in previous studies as candidate microorganisms for injection, and were used for this experiment. Oligonucleotide probes, design based on specific sequences in the 16S rRNA gene were labeled with either fluorescein isothiocyanate (FITC), or 6-car-boxy-X-rhodamine (ROX), and were allowed to hybridize with fixed cells of the two microorganisms noted above. The fluorescence signal emitted from each microorganism cells could clearly be detected by an epifluorescence microscope. Moreover, E. cloacae TRC-322 and B, licheniformis TRC-18-2-a, suspended in actual reservoir brine, including inorganic salts, oil and aboriginal cells of the reservoir brine, could be detected directly by this hybridization method, without the need for cultivation and isolation. (author)

  20. Phylogeny and FISH probe analysis of the “Candidatus Competibacter”-lineage in wastewater treatment systems

    DEFF Research Database (Denmark)

    Nittami, Tadashi; McIlroy, Simon Jon; Kanai, Eri

    Our understanding of the microbial ecology of enhanced biological phosphorus removal (EBPR) wastewater treatment systems has been greatly advanced through the application of molecular methods such as fluorescence in situ hybridization (FISH). Considerable attention has been directed at the identi......Our understanding of the microbial ecology of enhanced biological phosphorus removal (EBPR) wastewater treatment systems has been greatly advanced through the application of molecular methods such as fluorescence in situ hybridization (FISH). Considerable attention has been directed...... at the identification and characterization of the glycogen accumulating organisms (GAO), a phenotypic group thought to compete with the polyphosphate accumulating organisms (PAO) for resources at the theoretical expense of EBPR efficiency. Demonstrated candidates for members of the GAO phenotype include...... the gammaproteobacterial “Candidatus Competibacter”-lineage. The group is currently delineated by 8 FISH probe defined phylotypes, although further undescribed phylogenetic diversity beyond what is covered by these probes is evident. Where studied, marked differences in physiology between members are observed, including...

  1. Supernumerary ring chromosome 20 characterized by fluorescence in situ hybridization

    NARCIS (Netherlands)

    Van Langen, Irene M.; Otter, Mariëlle A.; Aronson, Daniël C.; Overweg-Plandsoen, W.C.G.; Hennekam, Raoul C.M.; Leschot, Nico J.; Hoovers, Jan M.N.

    1996-01-01

    We report on a boy with mild dysmorphic features and developmental delay, in whom karyotyping showed an additional minute ring chromosome in 60% of metaphases. Fluorescence in situ hybridization (FISH) with a centromere specific probe demonstrated that the ring chromosome contained the centromeric

  2. Some conclusions from shipboard experiments on the radurization of whole fish in the Federal Republic of Germany

    International Nuclear Information System (INIS)

    Ehlermann, D.A.E.; Reinacher, E.

    1978-01-01

    Redfish and haddock were radurized (100krad) with the X-ray facility onboard the research vessel ''Anton Dohrn''. The whole fish were stored under crushed ice. In addition whole haddock packaged in plastic pouches under vacuum were studied. In general, during the first 16 days of storage no marked difference in the sensory quality between irradiated and unirradiated samples was observed. Bacteriological and chemical analyses proved the effectiveness of radurization to reduce the microbial flora usually responsible for spoilage. When compared with the results from other shipboard experiments and laboratory studies, the results of sensory evaluation are also in general accordance - if the same criterion for the judgement of final quality is used. After a storage period longer than 16 days occasionally the irradiated samples were scored higher than the unirradiated ones, but then the quality was no longer acceptable. If a lower criterion for shelf-life extension is used, it may be stated that a considerable shelf-life extension results from irradiation. By the standards used in our experiments, it is not possible to preserve an acceptable quality and to extend the shelf-life of ocean fish under the conditions prevailing in practical fishing. Taking into account the many factors that influence the results of such studies, it is not excluded that under other conditions the beneficial effects of radiation processing may be observed. (author)

  3. Hybrid CNG propulsion for fleet vehicles: emission reduction potential and operating experience

    Energy Technology Data Exchange (ETDEWEB)

    Drozdz, P. [BC Research Institute, BC (Canada)

    1997-12-31

    A project (1) to build an experimental hybrid electric vehicle to be used as a test bed for the development of EZEV-oriented technologies, (2) to develop a control system to manage the energy use in a series hybrid vehicle, (3) to evaluate the suitability of valve regulated lead acid batteries for hybrid propulsion, and (4) to investigate the feasibility of using hybrid propulsion for medium duty fleet vehicles was discussed. In this context, the electric G-Van, the BCRI hybrid G-Van battery, the hybrid power unit, and the electronic control unit were described. The concept of hybrid vehicle control, and the control system software were explained, and a summary of the hybrid system efficiency test was provided.

  4. Fish allergens at a glance: variable allergenicity of parvalbumins, the major fish allergens.

    Science.gov (United States)

    Kuehn, Annette; Swoboda, Ines; Arumugam, Karthik; Hilger, Christiane; Hentges, François

    2014-01-01

    Fish is a common trigger of severe, food-allergic reactions. Only a limited number of proteins induce specific IgE-mediated immune reactions. The major fish allergens are the parvalbumins. They are members of the calcium-binding EF-hand protein family characterized by a conserved protein structure. They represent highly cross-reactive allergens for patients with specific IgE to conserved epitopes. These patients might experience clinical reactions with various fish species. On the other hand, some individuals have IgE antibodies directed against unique, species-specific parvalbumin epitopes, and these patients show clinical symptoms only with certain fish species. Furthermore, different parvalbumin isoforms and isoallergens are present in the same fish and might display variable allergenicity. This was shown for salmon homologs, where only a single parvalbumin (beta-1) isoform was identified as allergen in specific patients. In addition to the parvalbumins, several other fish proteins, enolases, aldolases, and fish gelatin, seem to be important allergens. New clinical and molecular insights advanced the knowledge and understanding of fish allergy in the last years. These findings were useful for the advancement of the IgE-based diagnosis and also for the management of fish allergies consisting of advice and treatment of fish-allergic patients.

  5. Fish allergens at a glance: Variable allergenicity of parvalbumins, the major fish allergens

    Directory of Open Access Journals (Sweden)

    Annette eKuehn

    2014-04-01

    Full Text Available Fish is a common trigger of severe, food-allergic reactions. Only a limited number of proteins induce specific IgE-mediated immune reactions. The major fish allergens are the parvalbumins. They are members of the calcium-binding EF-hand protein family characterized by a conserved protein structure. They represent highly cross-reactive allergens for patients with specific IgE to conserved epitopes. These patients might experience clinical reactions with various fish species. On the other hand, some individuals have IgE antibodies directed against unique, species-specific parvalbumin epitopes, and these patients show clinical symptoms only with certain fish species. Furthermore, different parvalbumin isoforms and isoallergens are present in the same fish and might display variable allergenicity. This was shown for salmon homologs, where only a single parvalbumin (beta-1 isoform was identified as allergen in specific patients. In addition to the parvalbumins, several other fish proteins, enolases, aldolases and fish gelatin, seem to be important allergens.New clinical and molecular insights advanced the knowledge and understanding of fish allergy in the last years. These findings will be useful for the advancement of the IgE-based diagnosis but also for the management of fish allergies consisting of advice and treatment of fish-allergic patients.

  6. Multicolor fluorescent in situ hybridization to define abutting and overlapping gene expression in the embryonic zebrafish brain

    Directory of Open Access Journals (Sweden)

    Hauptmann Giselbert

    2011-04-01

    Full Text Available Abstract Background In recent years, mapping of overlapping and abutting regulatory gene expression domains by chromogenic two-color in situ hybridization has helped define molecular subdivisions of the developing vertebrate brain and shed light on its basic organization. Despite the benefits of this technique, visualization of overlapping transcript distributions by differently colored precipitates remains difficult because of masking of lighter signals by darker color precipitates and lack of three-dimensional visualization properties. Fluorescent detection of transcript distributions may be able to solve these issues. However, despite the use of signal amplification systems for increasing sensitivity, fluorescent detection in whole-mounts suffers from rapid quenching of peroxidase (POD activity compared to alkaline phosphatase chromogenic reactions. Thus, less strongly expressed genes cannot be efficiently detected. Results We developed an optimized procedure for fluorescent detection of transcript distribution in whole-mount zebrafish embryos using tyramide signal amplification (TSA. Conditions for hybridization and POD-TSA reaction were optimized by the application of the viscosity-increasing polymer dextran sulfate and the use of the substituted phenol compounds 4-iodophenol and vanillin as enhancers of POD activity. In combination with highly effective bench-made tyramide substrates, these improvements resulted in dramatically increased signal-to-noise ratios. The strongly enhanced signal intensities permitted fluorescent visualization of less abundant transcripts of tissue-specific regulatory genes. When performing multicolor fluorescent in situ hybridization (FISH experiments, the highly sensitive POD reaction conditions required effective POD inactivation after each detection cycle by glycine-hydrochloric acid treatment. This optimized FISH procedure permitted the simultaneous fluorescent visualization of up to three unique transcripts

  7. Potential for bias in using hybrids between common carp (Cyprinus carpio) and goldfish (Carassius auratus) in endocrine studies: a first report of hybrids in Lake Mead, Nevada, U.S.A

    Science.gov (United States)

    Goodbred, Steven L.; Patino, Reynaldo; Orsak, Erik; Sharma, Prakash; Ruessler, Shane

    2013-01-01

    During a 2008 study to assess endocrine and reproductive health of common carp (Cyprinus carpio) in Lake Mead, Nevada (U.S.A.) we identified two fish, one male and one female, as hybrids with goldfish (Carassius auratus) based on morphology, lateral line scale count, and lack of anterior barbels. Gross examination of the female hybrid ovaries indicated presence of vitellogenic ovarian follicles; whereas histological evaluation of the male hybrid testes showed lobule-like structures with open lumens but without germ cells, suggesting it was sterile. Because common carp/goldfish hybrids are more susceptible to gonadal tumors and may have different endocrine profiles than common carp, researchers using common carp as a model for endocrine/reproductive studies should be aware of the possible presence of hybrids.

  8. Oral cancer/endothelial cell fusion experiences nuclear fusion and acquisition of enhanced survival potential.

    Science.gov (United States)

    Song, Kai; Song, Yong; Zhao, Xiao-Ping; Shen, Hui; Wang, Meng; Yan, Ting-Lin; Liu, Ke; Shang, Zheng-Jun

    2014-10-15

    Most previous studies have linked cancer-macrophage fusion with tumor progression and metastasis. However, the characteristics of hybrid cells derived from oral cancer and endothelial cells and their involvement in cancer remained unknown. Double-immunofluorescent staining and fluorescent in situ hybridization (FISH) were performed to confirm spontaneous cell fusion between eGFP-labeled human umbilical vein endothelial cells (HUVECs) and RFP-labeled SCC9, and to detect the expression of vementin and cytokeratin 18 in the hybrids. The property of chemo-resistance of such hybrids was examined by TUNEL assay. The hybrid cells in xenografted tumor were identified by FISH and GFP/RFP dual-immunofluoresence staining. We showed that SCC9 cells spontaneously fused with cocultured endothelial cells, and the resultant hybrid cells maintained the division and proliferation activity after re-plating and thawing. Such hybrids expressed markers of both parental cells and became more resistant to chemotherapeutic drug cisplatin as compared to the parental SCC9 cells. Our in vivo data indicated that the hybrid cells contributed to tumor composition by using of immunostaining and FISH analysis, even though the hybrid cells and SCC9 cells were mixed with 1:10,000, according to the FACS data. Our study suggested that the fusion events between oral cancer and endothelial cells undergo nuclear fusion and acquire a new property of drug resistance and consequently enhanced survival potential. These experimental findings provide further supportive evidence for the theory that cell fusion is involved in cancer progression. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Effects of salinity on gastric emptying time in hybrid grouper, Epinephelus fuscoguttatus x E. lanceolattus juveniles

    Science.gov (United States)

    Noor, Noorashikin Md.; Das, Simon Kumar; Cob, Zaidi Che; Ghaffar, Mazlan Abd.

    2018-04-01

    The newly developed hybrid grouper: tiger grouper (Epinephelus fuscoguttatus) × giant grouper (Epinephelus fuscoguttatus) (TG×GG), has a high resistance towards different environmental condition (eg. in euryhaline environment) due to its genetic improvement. This study aims to investigate the effects of different salinities (10, 15, 20, 25 and 30 ppt) on the gastric emptying time (GET) of the TG×GG hybrid grouper juveniles. The fish were fed with commercial pellet over a 30 days experimental period under controlled laboratory conditions. The GET was determined by X-radiographic method, using barium sulfate (BaSO4) as an inert food marker. The X-radiography images showed that the shortest GET (12 h) was observed in the 15 ppt group, whereas the longest GET (18 h) in 30 ppt group. The results suggests to culture TG×GG hybrid grouper juveniles in 15 ppt with commercial pellet diet as this salinity proliferates faster digestion process which may contribute faster growth rate of this important fish species. Overall, these findings would be useful for the betterment of TG×GG hybrid grouper aquaculture which will eventually boost up the production of this newly developed hybrid grouper species.

  10. Mate Choice Drives Evolutionary Stability in a Hybrid Complex.

    Directory of Open Access Journals (Sweden)

    Miguel Morgado-Santos

    Full Text Available Previous studies have shown that assortative mating acts as a driver of speciation by countering hybridization between two populations of the same species (pre-zygotic isolation or through mate choice among the hybrids (hybrid speciation. In both speciation types, assortative mating promotes speciation over a transient hybridization stage. We studied mate choice in a hybrid vertebrate complex, the allopolyploid fish Squalius alburnoides. This complex is composed by several genomotypes connected by an intricate reproductive dynamics. We developed a model that predicts the hybrid complex can persist when females exhibit particular mate choice patterns. Our model is able to reproduce the diversity of population dynamic outcomes found in nature, namely the dominance of the triploids and the dominance of the tetraploids, depending on female mate choice patterns and frequency of the parental species. Experimental mate choice trials showed that females exhibit the preferences predicted by the model. Thus, despite the known role of assortative mating in driving speciation, our findings suggest that certain mate choice patterns can instead hinder speciation and support the persistence of hybrids over time without speciation or extinction.

  11. Oral cancer/endothelial cell fusion experiences nuclear fusion and acquisition of enhanced survival potential

    International Nuclear Information System (INIS)

    Song, Kai; Song, Yong; Zhao, Xiao-Ping; Shen, Hui; Wang, Meng; Yan, Ting-lin; Liu, Ke; Shang, Zheng-jun

    2014-01-01

    Most previous studies have linked cancer–macrophage fusion with tumor progression and metastasis. However, the characteristics of hybrid cells derived from oral cancer and endothelial cells and their involvement in cancer remained unknown. Double-immunofluorescent staining and fluorescent in situ hybridization (FISH) were performed to confirm spontaneous cell fusion between eGFP-labeled human umbilical vein endothelial cells (HUVECs) and RFP-labeled SCC9, and to detect the expression of vementin and cytokeratin 18 in the hybrids. The property of chemo-resistance of such hybrids was examined by TUNEL assay. The hybrid cells in xenografted tumor were identified by FISH and GFP/RFP dual-immunofluoresence staining. We showed that SCC9 cells spontaneously fused with cocultured endothelial cells, and the resultant hybrid cells maintained the division and proliferation activity after re-plating and thawing. Such hybrids expressed markers of both parental cells and became more resistant to chemotherapeutic drug cisplatin as compared to the parental SCC9 cells. Our in vivo data indicated that the hybrid cells contributed to tumor composition by using of immunostaining and FISH analysis, even though the hybrid cells and SCC9 cells were mixed with 1:10,000, according to the FACS data. Our study suggested that the fusion events between oral cancer and endothelial cells undergo nuclear fusion and acquire a new property of drug resistance and consequently enhanced survival potential. These experimental findings provide further supportive evidence for the theory that cell fusion is involved in cancer progression. - Highlights: • The fusion events between oral cancer and endothelial cells undergo nuclear fusion. • The resulting hybrid cells acquire a new property of drug resistance. • The resulting hybrid cells express the markers of both parental cells (i.e. vimentin and cytokeratin 18). • The hybrid cells contribute to tumor repopulation in vivo

  12. Oral cancer/endothelial cell fusion experiences nuclear fusion and acquisition of enhanced survival potential

    Energy Technology Data Exchange (ETDEWEB)

    Song, Kai [Department of Oral and Maxillofacial Surgery, The Affiliated Hospital of Qingdao University, Shandong Province (China); The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory for Oral Biomedicine Ministry of Education, Wuhan University, Wuhan (China); Song, Yong [The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory for Oral Biomedicine Ministry of Education, Wuhan University, Wuhan (China); Department of Stomatology, Liu Zhou People' s Hospital, Guangxi (China); Zhao, Xiao-Ping; Shen, Hui; Wang, Meng; Yan, Ting-lin [The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory for Oral Biomedicine Ministry of Education, Wuhan University, Wuhan (China); Liu, Ke, E-mail: liuke.1999@aliyun.com [Department of Oral and Maxillofacial-Head and Neck oncology, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079 (China); The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory for Oral Biomedicine Ministry of Education, Wuhan University, Wuhan (China); Shang, Zheng-jun, E-mail: shangzhengjun@hotmail.com [Department of Oral and Maxillofacial-Head and Neck oncology, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079 (China); The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory for Oral Biomedicine Ministry of Education, Wuhan University, Wuhan (China)

    2014-10-15

    Most previous studies have linked cancer–macrophage fusion with tumor progression and metastasis. However, the characteristics of hybrid cells derived from oral cancer and endothelial cells and their involvement in cancer remained unknown. Double-immunofluorescent staining and fluorescent in situ hybridization (FISH) were performed to confirm spontaneous cell fusion between eGFP-labeled human umbilical vein endothelial cells (HUVECs) and RFP-labeled SCC9, and to detect the expression of vementin and cytokeratin 18 in the hybrids. The property of chemo-resistance of such hybrids was examined by TUNEL assay. The hybrid cells in xenografted tumor were identified by FISH and GFP/RFP dual-immunofluoresence staining. We showed that SCC9 cells spontaneously fused with cocultured endothelial cells, and the resultant hybrid cells maintained the division and proliferation activity after re-plating and thawing. Such hybrids expressed markers of both parental cells and became more resistant to chemotherapeutic drug cisplatin as compared to the parental SCC9 cells. Our in vivo data indicated that the hybrid cells contributed to tumor composition by using of immunostaining and FISH analysis, even though the hybrid cells and SCC9 cells were mixed with 1:10,000, according to the FACS data. Our study suggested that the fusion events between oral cancer and endothelial cells undergo nuclear fusion and acquire a new property of drug resistance and consequently enhanced survival potential. These experimental findings provide further supportive evidence for the theory that cell fusion is involved in cancer progression. - Highlights: • The fusion events between oral cancer and endothelial cells undergo nuclear fusion. • The resulting hybrid cells acquire a new property of drug resistance. • The resulting hybrid cells express the markers of both parental cells (i.e. vimentin and cytokeratin 18). • The hybrid cells contribute to tumor repopulation in vivo.

  13. Identification of MYCN gene amplification in neuroblastoma using chromogenic in situ hybridization (CISH): an alternative and practical method.

    Science.gov (United States)

    Bhargava, Rohit; Oppenheimer, Orit; Gerald, William; Jhanwar, Suresh C; Chen, Beiyun

    2005-06-01

    Chromogenic in situ hybridization (CISH) is a recently developed technique, which utilizes the general principles of in situ hybridization and a detection system similar to immunohistochemistry. To assess the utility of CISH for analysis of MYCN gene amplification, we compared this assay with established diagnostic assays such as Southern blot analysis (SB) and fluorescent in situ hybridization (FISH). CISH was performed on 67 cases of neuroblastoma using tissue microarray (65 cases) and whole tissue sections (2 cases). Unequivocal, high-level amplification (> or =10 gene copies per tumor nucleus) was identified in 19 of 67 (28.4%) tumors. Two (3%) tumors showed low-level amplification (6-9 gene copies per tumor nucleus). No amplification was seen in 46 of 67 (68.6%) tumors. SB data were available in 44 tumors. Forty-one of the 44 tumors (93%) showed concordant results between CISH and SB. Three tumors showed MYCN amplification by CISH but no amplification by SB, most likely due to dilution effect of nonneoplastic tissue in the test samples. Two of these three tumors also showed MYCN amplification by FISH, and the third tumor was not analyzed by FISH. FISH data were available in total of 30 tumors. All 30 tumors showed concordant results between CISH and FISH for classifying a tumor as MYCN amplified or not amplified. We conclude that CISH is an accurate method for determining MYCN gene amplification, with added advantages that make it a more practically useful method.

  14. Genetic investigation of natural hybridization between rainbow and coastal cutthroat trout in the copper River Delta, Alaska

    Science.gov (United States)

    Williams, I.; Reeves, G.H.; Graziano, S.L.; Nielsen, J.L.

    2007-01-01

    Molecular genetic methods were used to quantify natural hybridization between rainbow trout Oncorhynchus mykiss or steelhead (anadromous rainbow trout) and coastal cutthroat trout O. clarkii clarkii collected in the Copper River delta, Southeast Alaska. Eleven locations were sampled to determine the extent of hybridization and the distribution of hybrids. Four diagnostic nuclear microsatellite loci and four species-specific simple sequence repeat markers were used in combination with restriction fragment length polymorphism analyses of NADH dehydrogenase 5/6 (ND5/6) mitochondrial DNA (mtDNA) to investigate the genetic structure of trout from both species and identify putative interspecific hybrids. Hybrids were found in 7 of the 11 streams sampled in the Copper River delta, the extent of hybridization across all streams varying from 0% to 58%. Hybrid trout distribution appeared to be nonrandom, most individuals of mixed taxonomic ancestry being detected in streams containing rainbow trout rather than in streams containing coastal cutthroat trout. Genotypic disequilibrium was observed among microsatellite loci in populations with high levels of hybridization. We found no significant correlation between unique stream channel process groups and the number of hybrid fish sampled. Eighty-eight percent of fish identified as first-generation hybrids (F1) in two populations contained coastal cutthroat trout mtDNA, suggesting directionality in hybridization. However, dominance of coastal cutthroat trout mtDNA was not observed at a third location containing F1 hybrids, indicating that interspecific mating behavior varied among locations. Backcrossed individuals were found in drainages lacking F1 hybrids and in populations previously thought to contain a single species. The extent and distribution of backcrossed individuals suggested that at least some hybrids are reproductively viable and backcrossed hybrid offspring move throughout the system.

  15. Behavioural isolation may facilitate homoploid hybrid speciation in cichlid fish

    NARCIS (Netherlands)

    Selz, O. M.; Thommen, R.; Maan, M. E.; Seehausen, O.

    Hybrid speciation is constrained by the homogenizing effects of gene flow from the parental species. In the absence of post-mating isolation due to structural changes in the genome, or temporal or spatial premating isolation, another form of reproductive isolation would be needed for homoploid

  16. Intelligent Fish Freshness Assessment

    Directory of Open Access Journals (Sweden)

    Hamid Gholam Hosseini

    2008-01-01

    Full Text Available Fish species identification and automated fish freshness assessment play important roles in fishery industry applications. This paper describes a method based on support vector machines (SVMs to improve the performance of fish identification systems. The result is used for the assessment of fish freshness using artificial neural network (ANN. Identification of the fish species involves processing of the images of fish. The most efficient features were extracted and combined with the down-sampled version of the images to create a 1D input vector. Max-Win algorithm applied to the SVM-based classifiers has enhanced the reliability of sorting to 96.46%. The realisation of Cyranose 320 Electronic nose (E-nose, in order to evaluate the fish freshness in real-time, is experimented. Intelligent processing of the sensor patterns involves the use of a dedicated ANN for each species under study. The best estimation of freshness was provided by the most sensitive sensors. Data was collected from four selected species of fishes over a period of ten days. It was concluded that the performance can be increased using individual trained ANN for each specie. The proposed system has been successful in identifying the number of days after catching the fish with an accuracy of up to 91%.

  17. Automated Image Analysis for Quantitative Fluorescence In Situ Hybridization with Environmental Samples▿ †

    OpenAIRE

    Zhou, Zhi; Pons, Marie Noëlle; Raskin, Lutgarde; Zilles, Julie L.

    2007-01-01

    When fluorescence in situ hybridization (FISH) analyses are performed with complex environmental samples, difficulties related to the presence of microbial cell aggregates and nonuniform background fluorescence are often encountered. The objective of this study was to develop a robust and automated quantitative FISH method for complex environmental samples, such as manure and soil. The method and duration of sample dispersion were optimized to reduce the interference of cell aggregates. An au...

  18. Phytoplankton IF-FISH: Species-specific labeling of cellular proteins by immunofluorescence (IF) with simultaneous species identification by fluorescence immunohybridization (FISH).

    Science.gov (United States)

    Meek, Megan E; Van Dolah, Frances M

    2016-05-01

    Phytoplankton rarely occur as unialgal populations. Therefore, to study species-specific protein expression, indicative of physiological status in natural populations, methods are needed that will both assay for a protein of interest and identify the species expressing it. Here we describe a protocol for IF-FISH, a dual labeling procedure using immunofluorescence (IF) labeling of a protein of interest followed by fluorescence in situ hybridization (FISH) to identify the species expressing that protein. The protocol was developed to monitor expression of the cell cycle marker proliferating cell nuclear antigen (PCNA) in the red tide dinoflagellate, Karenia brevis, using a large subunit (LSU) rRNA probe to identify K. brevis in a mixed population of morphologically similar Karenia species. We present this protocol as proof of concept that IF-FISH can be successfully applied to phytoplankton cells. This method is widely applicable for the analysis of single-cell protein expression of any protein of interest within phytoplankton communities. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. A hybrid genetic linkage map of two ecologically and morphologically divergent Midas cichlid fishes (Amphilophus spp.) obtained by massively parallel DNA sequencing (ddRADSeq).

    Science.gov (United States)

    Recknagel, Hans; Elmer, Kathryn R; Meyer, Axel

    2013-01-01

    Cichlid fishes are an excellent model system for studying speciation and the formation of adaptive radiations because of their tremendous species richness and astonishing phenotypic diversity. Most research has focused on African rift lake fishes, although Neotropical cichlid species display much variability as well. Almost one dozen species of the Midas cichlid species complex (Amphilophus spp.) have been described so far and have formed repeated adaptive radiations in several Nicaraguan crater lakes. Here we apply double-digest restriction-site associated DNA sequencing to obtain a high-density linkage map of an interspecific cross between the benthic Amphilophus astorquii and the limnetic Amphilophus zaliosus, which are sympatric species endemic to Crater Lake Apoyo, Nicaragua. A total of 755 RAD markers were genotyped in 343 F(2) hybrids. The map resolved 25 linkage groups and spans a total distance of 1427 cM with an average marker spacing distance of 1.95 cM, almost matching the total number of chromosomes (n = 24) in these species. Regions of segregation distortion were identified in five linkage groups. Based on the pedigree of parents to F(2) offspring, we calculated a genome-wide mutation rate of 6.6 × 10(-8) mutations per nucleotide per generation. This genetic map will facilitate the mapping of ecomorphologically relevant adaptive traits in the repeated phenotypes that evolved within the Midas cichlid lineage and, as the first linkage map of a Neotropical cichlid, facilitate comparative genomic analyses between African cichlids, Neotropical cichlids and other teleost fishes.

  20. Application of Pseudomurein Endoisopeptidase to Fluorescence In Situ Hybridization of Methanogens within the Family Methanobacteriaceae▿

    Science.gov (United States)

    Nakamura, Kohei ; Terada, Takeshi; Sekiguchi, Yuji; Shinzato, Naoya; Meng, Xian-Ying; Enoki, Miho; Kamagata, Yoichi

    2006-01-01

    In situ detection of methanogens within the family Methanobacteriaceae is sometimes known to be unsuccessful due to the difficulty in permeability of oligonucleotide probes. Pseudomurein endoisopeptidase (Pei), a lytic enzyme that specifically acts on their cell walls, was applied prior to 16S rRNA-targeting fluorescence in situ hybridization (FISH). For this purpose, pure cultured methanogens within this family, Methanobacterium bryantii, Methanobrevibacter ruminantium, Methanosphaera stadtmanae, and Methanothermobacter thermautotrophicus together with a Methanothermobacter thermautotrophicus-containing syntrophic acetate-oxidizing coculture, endosymbiotic Methanobrevibacter methanogens within an anaerobic ciliate, and an upflow anaerobic sludge blanket (UASB) granule were examined. Even without the Pei treatment, Methanobacterium bryantii and Methanothermobacter thermautotrophicus cells are relatively well hybridized with oligonucleotide probes. However, almost none of the cells of Methanobrevibacter ruminantium, Methanosphaera stadtmanae, cocultured Methanothermobacter thermautotrophicus, and the endosymbiotic methanogens and the cells within UASB granule were hybridized. Pei treatment was able to increase the probe hybridization ratio in every specimen, particularly in the specimen that had shown little hybridization. Interestingly, the hybridizing signal intensity of Methanothermobacter thermautotrophicus cells in coculture with an acetate-oxidizing H2-producing syntroph was significantly improved by Pei pretreatment, whereas the probe was well hybridized with the cells of pure culture of the same strain. We found that the difference is attributed to the differences in cell wall thicknesses between the two culture conditions. These results indicate that Pei treatment is effective for FISH analysis of methanogens that show impermeability to the probe. PMID:16950902

  1. Analysis of the Ceratitis capitata y chromosome using in situ hybridization to mitotic chromosomes

    International Nuclear Information System (INIS)

    Willhoeft, U.; Franz, G.

    1998-01-01

    In Ceratitis capitata the Y chromosome is responsible for sex-determination. We used fluorescence in situ hybridization (FISH) for cytogenetic analysis of mitotic chromosomes. FISH with the wild-type strain EgyptII and two repetitive DNA probes enabled us to differentiate between the short and the long arm of the Y chromosome and gives a much better resolution than C-banding of mitotic chromosomes. We identified the Y-chromosomal breakpoints in Y-autosome translocations using FISH. Even more complex rearrangements i.e. deletions and insertions in some translocation strains were detected by this method. A strategy for mapping the primary sex determination factor in Ceratitis capitata by FISH is presented. (author)

  2. Automated processing of fluorescence in-situ hybridization slides for HER2 testing in breast and gastro-esophageal carcinomas.

    Science.gov (United States)

    Tafe, Laura J; Allen, Samantha F; Steinmetz, Heather B; Dokus, Betty A; Cook, Leanne J; Marotti, Jonathan D; Tsongalis, Gregory J

    2014-08-01

    HER2 fluorescence in-situ hybridization (FISH) is used in breast and gastro-esophageal carcinoma for determining HER2 gene amplification and patients' eligibility for HER2 targeted therapeutics. Traditional manual processing of the FISH slides is labor intensive because of multiple steps that require hands on manipulation of the slides and specifically timed intervals between steps. This highly manual processing also introduces inter-run and inter-operator variability that may affect the quality of the FISH result. Therefore, we sought to incorporate an automated processing instrument into our FISH workflow. Twenty-six cases including breast (20) and gastro-esophageal (6) cancer comprising 23 biopsies and three excision specimens were tested for HER2 FISH (Pathvysion, Abbott) using the Thermobrite Elite (TBE) system (Leica). Up to 12 slides can be run simultaneously. All cases were previously tested by the Pathvysion HER2 FISH assay with manual preparation. Twenty cells were counted by two observers for each case; five cases were tested on three separate runs by different operators to evaluate the precision and inter-operator variability. There was 100% concordance in the scoring between the manual and TBE methods as well as among the five cases that were tested on three runs. Only one case failed due to poor probe hybridization. In total, seven cases were positive for HER2 amplification (HER2:CEP17 ratio >2.2) and the remaining 19 were negative (HER2:CEP17 ratio <1.8) utilizing the 2007 ASCO/CAP scoring criteria. Due to the automated denaturation and hybridization, for each run, there was a reduction in labor of 3.5h which could then be dedicated to other lab functions. The TBE is a walk away pre- and post-hybridization system that automates FISH slide processing, improves work flow and consistency and saves approximately 3.5h of technologist time. The instrument has a small footprint thus occupying minimal counter space. TBE processed slides performed

  3. EGFR mutation is a better predictor of response to tyrosine kinase inhibitors in non-small cell lung carcinoma than FISH, CISH, and immunohistochemistry.

    Science.gov (United States)

    Sholl, Lynette M; Xiao, Yun; Joshi, Victoria; Yeap, Beow Y; Cioffredi, Leigh-Anne; Jackman, David M; Lee, Charles; Jänne, Pasi A; Lindeman, Neal I

    2010-06-01

    About 10% of patients with non-small cell lung carcinoma (NSCLC) respond to epidermal growth factor receptor (EGFR)-targeted tyrosine kinase inhibitors (TKIs). More than 75% of "responders" have activating mutations in EGFR. However, mutation analysis is not widely available, and proposed alternatives (in situ hybridization and immunohistochemical analysis) have shown inconsistent associations with outcome. Fluorescence in situ hybridization (FISH), chromogenic in situ hybridization (CISH), immunohistochemical analysis, and DNA sequencing were compared in this study of 40 NSCLC samples from TKI-treated patients. Response rates were 12 of 19 in EGFR-mutant vs 1 of 20 EGFR wild-type tumors (P = .0001), 7 of 19 FISH+ vs 4 of 17 FISH- tumors (not significant [NS]), 5 of 16 CISH+ vs 6 of 21 CISH- tumors (NS), and 3 of 9 immunohistochemically positive vs 7 of 22 immunohistochemically negative tumors (NS). EGFR mutation was associated with improved progression-free survival (P = .0004). Increased copy number (FISH or CISH) and protein expression (immunohistochemical) did not independently predict outcome. Thus, EGFR sequence analysis was the only method useful for predicting response and progression-free survival following TKI therapy in NSCLC.

  4. Seven-hour fluorescence in situ hybridization technique for enumeration of Enterobacteriaceae in food and environmental water sample.

    Science.gov (United States)

    Ootsubo, M; Shimizu, T; Tanaka, R; Sawabe, T; Tajima, K; Ezura, Y

    2003-01-01

    A fluorescent in situ hybridization (FISH) technique using an Enterobacteriaceae-specific probe (probe D) to target 16S rRNA was improved in order to enumerate, within a single working day, Enterobacteriaceae present in food and environmental water samples. In order to minimize the time required for the FISH procedure, each step of FISH with probe D was re-evaluated using cultured Escherichia coli. Five minutes of ethanol treatment for cell fixation and hybridization were sufficient to visualize cultured E. coli, and FISH could be performed within 1 h. Because of the difficulties in detecting low levels of bacterial cells by FISH without cultivation, a FISH technique for detecting microcolonies on membrane filters was investigated to improve the bacterial detection limit. FISH with probe D following 6 h of cultivation to grow microcolonies on a 13 mm diameter membrane filter was performed, and whole Enterobacteriaceae microcolonies on the filter were then detected and enumerated by manual epifluorescence microscopic scanning at magnification of x100 in ca 5 min. The total time for FISH with probe D following cultivation (FISHFC) was reduced to within 7 h. FISHFC can be applied to enumerate cultivable Enterobacteriaceae in food (above 100 cells g-1) and environmental water samples (above 1 cell ml-1). Cultivable Enterobacteriaceae in food and water samples were enumerated accurately within 7 h using the FISHFC method. A FISHFC method capable of evaluating Enterobacteriaceae contamination in food and environmental water within a single working day was developed.

  5. Multiple cues produced by a robotic fish modulate aggressive behaviour in Siamese fighting fishes.

    Science.gov (United States)

    Romano, Donato; Benelli, Giovanni; Donati, Elisa; Remorini, Damiano; Canale, Angelo; Stefanini, Cesare

    2017-07-05

    The use of robotics to establish social interactions between animals and robots, represents an elegant and innovative method to investigate animal behaviour. However, robots are still underused to investigate high complex and flexible behaviours, such as aggression. Here, Betta splendens was tested as model system to shed light on the effect of a robotic fish eliciting aggression. We evaluated how multiple signal systems, including a light stimulus, affect aggressive responses in B. splendens. Furthermore, we conducted experiments to estimate if aggressive responses were triggered by the biomimetic shape of fish replica, or whether any intruder object was effective as well. Male fishes showed longer and higher aggressive displays as puzzled stimuli from the fish replica increased. When the fish replica emitted its full sequence of cues, the intensity of aggression exceeded even that produced by real fish opponents. Fish replica shape was necessary for conspecific opponent perception, evoking significant aggressive responses. Overall, this study highlights that the efficacy of an artificial opponent eliciting aggressive behaviour in fish can be boosted by exposure to multiple signals. Optimizing the cue combination delivered by the robotic fish replica may be helpful to predict escalating levels of aggression.

  6. FISH as A method for detection of radiation Induced genetic damage

    International Nuclear Information System (INIS)

    Lakatosova, M.; Holeckova, B.

    2006-01-01

    Fluorescence in situ hybridization (FISH) has been considered as a suitable method for rapid and easy detection of chromosome aberrations. In contrast to the standard conventional staining procedure, this technique enables the detection and specification of stable chromosomal re-arrangements, which are compatible with cellular division and thus, they could be transmitted from common ancestral to next cell generations. FISH chromosome - specific painting probes have been effectively applied for the detection of chromosomal damage after exposure to radiation. During last years, several specific fluorescent labeled probes were performed that allowed precise detection of centromeres, sub-telomeres or other regions (sequences) in genome. Our paper deals with describing of different types of FISH probes and their possibilities for application in radiobiology. (authors)

  7. Fluorescence in situ hybridization and qPCR to detect Merkel cell polyomavirus physical status and load in Merkel cell carcinomas.

    Science.gov (United States)

    Haugg, Anke M; Rennspiess, Dorit; zur Hausen, Axel; Speel, Ernst-Jan M; Cathomas, Gieri; Becker, Jürgen C; Schrama, David

    2014-12-15

    The Merkel cell polyomavirus (MCPyV) is detected in 80% of Merkel cell carcinomas (MCC). Clonal integration and tumor-specific mutations in the large T antigen are strong arguments that MCPyV is a human tumor virus. However, the relationship between viral presence and cancer induction remains discussed controversially. Since almost all studies on virus prevalence are based on PCR techniques, we performed MCPyV fluorescence in situ hybridization (FISH) on MCC to gain information about the quality of the viral presence on the single cell level. MCPyV-FISH was performed on tissue microarrays containing 62 formalin-fixed and paraffin-embedded tissue samples including all tumor grades of 42 patients. The hybridization patterns were correlated to the qPCR data determined on corresponding whole tissue sections. Indeed, MCPyV-FISH and qPCR data were highly correlated, i.e. 83% for FISH-positive and 93% for FISH-negative cores. Accordingly, the mean of the qPCR values of all MCPyV-positive cores differed significantly from the mean of the negative cores (p = 0.0076). Importantly, two hybridization patterns were definable in the MCPyV-FISH: a punctate pattern (85%) indicating viral integration, which correlated with a moderate viral abundance and a combination of the punctate with a diffuse pattern (15%), suggesting a possible coexistence of integrated and episomal virus which was associated with very high viral load and VP1 expression. Thus, MCPyV-FISH adds important information on the single cell level within the histomorphological context and could therefore be an important tool to further elucidate MCPyV related carcinogenesis. © 2014 UICC.

  8. Detection of ALK gene rearrangement in non-small cell lung cancer: a comparison of fluorescence in situ hybridization and chromogenic in situ hybridization with correlation of ALK protein expression.

    Science.gov (United States)

    Kim, Hyojin; Yoo, Seol-Bong; Choe, Ji-Young; Paik, Jin Ho; Xu, Xianhua; Nitta, Hiroaki; Zhang, Wenjun; Grogan, Thomas M; Lee, Choon-Taek; Jheon, Sanghoon; Chung, Jin-Haeng

    2011-08-01

    Accurate determination of ALK rearrangement is important in lung cancer patients, especially in determining their eligibility for crizotinib therapy. Fluorescence in situ hybridization (FISH) has been regarded as the gold standard method for detecting ALK rearrangement. However, FISH requires a fluorescence microscope, and the signals are labile and rapidly fade over time. This study evaluates the concordance between ALK gene rearrangement in non-small cell lung cancer assessed by ALK FISH and a newly developed ALK chromogenic in situ hybridization (CISH) and correlates the results with ALK protein expression assessed by immunohistochemistry. A total of 465 formalin-fixed, paraffin-embedded non-small cell lung cancer samples were analyzed by ALK FISH (PathVysion, Vysis, Abbott) and ALK CISH. For comparison, all specimens were stained by immunohistochemistry (clone 5A4, Novocastra) and interobserver reproducibility was assessed. We found that agreement between the pathologists on the CISH-determined ALK status was achieved in 449 patients (96.6%), and ALK rearrangement was identified in 18 patients (4.0%) in CISH method. Among these cases, 443 cases (95.3%) had results matching the corresponding FISH results: 17 rearranged, 425 wild types, and 1 discordant case. There was high concordance in the assessment of ALK gene rearrangement between FISH and CISH techniques (κ = 0.92) and between observers (κ = 0.97). In addition, there was high concordance in the ALK gene status and ALK protein expression between CISH and IHC tests (κ = 0.82). CISH is a highly reproducible and practical method to detect ALK gene rearrangement and correlated well with ALK protein expression. Here, we present a diagnostic algorithm (Chung's SNUBH ALK protocol) to detect lung cancer with ALK rearrangements using IHC, FISH and CISH. Because CISH allows a concurrent analysis of histological features of the tumors and gene rearrangement, it appears to be a useful method in determining ALK gene

  9. RNA Imaging with Multiplexed Error Robust Fluorescence in situ Hybridization

    Science.gov (United States)

    Moffitt, Jeffrey R.; Zhuang, Xiaowei

    2016-01-01

    Quantitative measurements of both the copy number and spatial distribution of large fractions of the transcriptome in single-cells could revolutionize our understanding of a variety of cellular and tissue behaviors in both healthy and diseased states. Single-molecule Fluorescence In Situ Hybridization (smFISH)—an approach where individual RNAs are labeled with fluorescent probes and imaged in their native cellular and tissue context—provides both the copy number and spatial context of RNAs but has been limited in the number of RNA species that can be measured simultaneously. Here we describe Multiplexed Error Robust Fluorescence In Situ Hybridization (MERFISH), a massively parallelized form of smFISH that can image and identify hundreds to thousands of different RNA species simultaneously with high accuracy in individual cells in their native spatial context. We provide detailed protocols on all aspects of MERFISH, including probe design, data collection, and data analysis to allow interested laboratories to perform MERFISH measurements themselves. PMID:27241748

  10. Identification of hemiclonal reproduction in three species of Hexagrammos marine reef fishes.

    Science.gov (United States)

    Kimura-Kawaguchi, M R; Horita, M; Abe, S; Arai, K; Kawata, M; Munehara, H

    2014-08-01

    Natural hybrids between the boreal species Hexagrammos octogrammus and two temperate species Hexagrammos agrammus and Hexagrammos otakii were observed frequently in southern Hokkaido, Japan. Previous studies revealed that H. octogrammus is a maternal ancestor of both hybrids; the hybrids are all fertile females and they frequently breed with paternal species. Although such rampant hybridization occurs, species boundaries have been maintained in the hybrid zone. Possible explanations for the absence of introgressions, despite the frequent backcrossing, might include clonal reproduction: parthenogenesis, gynogenesis and hybridogenesis. The natural hybrids produced haploid eggs that contained only the H. octogrammus genome (maternal ancestor) with discarded paternal genome and generated F1 -hybrid type offspring by fertilization with the haploid sperm of H. agrammus or H. otakii (paternal ancestor). This reproductive mode was found in an artificial backcross hybrid between the natural hybrid and a male of the paternal ancestor. These findings indicate that the natural hybrids adopt hybridogenesis with high possibility and produce successive generations through hybridogenesis by backcrossing with the paternal ancestor. These hybrids of Hexagrammos represent the first hybridogenetic system found from marine fishes that widely inhabit the North Pacific Ocean. In contrast with other hybridogenetic systems, these Hexagrammos hybrids coexist with all three ancestral species in the hybrid zone. The coexistence mechanism is also discussed. © 2014 The Fisheries Society of the British Isles.

  11. Large-scale polymorphism near the ends of several human chromosomes analyzed by using fluorescence in situ hybridization (FISH)

    Energy Technology Data Exchange (ETDEWEB)

    Trask, B.J.; Friedman, C. [Univ. of Washington, Seattle, WA (United States); Giorgi, D. [CNRS, Montpelier (France)] [and others

    1994-09-01

    We have discovered a large DNA segment that is polymorphically present at the ends of several human chromosomes. The segment, f7501, was originally derived form a human chromosome 19-specific cosmid library. FISH was used to determine the cosmid`s chromosomal distribution on 44 unrelated humans and several closely related primates. The human subjects represent a diversity of reproductively isolated ethnic populations. FISH analysis revealed that sequences highly homologous to the cosmid`s insert are present on both homologs at 3q, 15q,. and 19p in almost all individuals (88, 85, and 87 of 88 homologs, respectively). Other chromosomes sites were labeled much more rarely in the sampled individuals. For example, 56 of the 88 analyzed chromosomes 11 were labeled (18+/+, 6-/-, and 20+/- individuals). In contrast, 2q was labeled on only 1/88 sampled chromosomes. The termini of 2q, 5q, 6p, 6q, 7p, 8p, 9p, 9q, 11p, 12q, 16p, 19q, and 20q and an interstitial site at 2q13-14 were labeled in at least one individual of the set. EcoR1-fragments derived from the cosmid showed the same hybridization pattern as the entire cosmid, indicating that at least 40 kbp is shared by these chromosome ends. Ethnic differences in the allele frequency of these polymorphic variants was observed. For example, signals were observed on 8/10 and 7/10 of the chromosomes 7p and 16q, respectively, derived form Biakan Pygmies, but these sites were infrequently labeled in non-Pygmy human populations (2/68, respectively). This region has undergone significant changes in chromosome location during human evolution. Strong signal was seen on chimpanzee and gorilla chromosome 3, which is homologous to human chromosome 4, a chromosome unlabeled in any of the humans we have analyzed.

  12. A comparative genomic hybridization study in a 46,XX male.

    Science.gov (United States)

    Rigola, M Angels; Carrera, Marta; Ribas, Isabel; Egozcue, Josep; Miró, Rosa; Fuster, Carme

    2002-07-01

    To identify Y chromosome material in an azoospermic male with an XX karyotype. Case report. Faculty of medicine and Centro de Patologia Celular (CPC) medical center. A 33-year-old man with infertility. G-banding, fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR), and comparative genomic hybridization (CGH). FISH for X and Y chromosomes, PCR for the SRYgene and amelogenin gene in the Xp (AMGX) and (AMGY), and losses or gains with CGH. FISH analysis using X and Y chromosome-specific probes showed an X chromosome containing Y chromosome sequences on the top of the short arm; this Y chromosome region was not visible by conventional cytogenetic analysis. PCR amplification of DNA showed the presence of the sex-determining region of the Y chromosome (SRY) and the amelogenin gene in the pseudoautosomal boundary of the X chromosome (AMGX). CGH confirmed the presence of the chromosome region Yp11.2-pter and detected the presence of the two otherwise normal X chromosomes. The two Xpter (XPAR1) pseudoautosomal regions present in this XX male suggest the need to reevaluate XX males using CGH and PCR to characterize the clinical variability in XX males due to genes other than those located on the Y chromosome.

  13. Monosomy 3 by FISH in uveal melanoma: variability in techniques and results.

    Science.gov (United States)

    Aronow, Mary; Sun, Yang; Saunthararajah, Yogen; Biscotti, Charles; Tubbs, Raymond; Triozzi, Pierre; Singh, Arun D

    2012-09-01

    Tumor monosomy 3 confers a poor prognosis in patients with uveal melanoma. We critically review the techniques used for fluorescence in situ hybridization (FISH) detection of monosomy 3 in order to assess variability in practice patterns and to explain differences in results. Significant variability that has likely affected reported results was found in tissue sampling methods, selection of FISH probes, number of cells counted, and the cut-off point used to determine monosomy 3 status. Clinical parameters and specific techniques employed to report FISH results should be specified so as to allow meta-analysis of published studies. FISH-based detection of monosomy 3 in uveal melanoma has not been performed in a standardized manner, which limits conclusions regarding its clinical utility. FISH is a widely available, versatile technology, and when performed optimally has the potential to be a valuable tool for determining the prognosis of uveal melanoma. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. Learning the scientific method using GloFish.

    Science.gov (United States)

    Vick, Brianna M; Pollak, Adrianna; Welsh, Cynthia; Liang, Jennifer O

    2012-12-01

    Here we describe projects that used GloFish, brightly colored, fluorescent, transgenic zebrafish, in experiments that enabled students to carry out all steps in the scientific method. In the first project, students in an undergraduate genetics laboratory course successfully tested hypotheses about the relationships between GloFish phenotypes and genotypes using PCR, fluorescence microscopy, and test crosses. In the second and third projects, students doing independent research carried out hypothesis-driven experiments that also developed new GloFish projects for future genetics laboratory students. Brianna Vick, an undergraduate student, identified causes of the different shades of color found in orange GloFish. Adrianna Pollak, as part of a high school science fair project, characterized the fluorescence emission patterns of all of the commercially available colors of GloFish (red, orange, yellow, green, blue, and purple). The genetics laboratory students carrying out the first project found that learning new techniques and applying their knowledge of genetics were valuable. However, assessments of their learning suggest that this project was not challenging to many of the students. Thus, the independent projects will be valuable as bases to widen the scope and range of difficulty of experiments available to future genetics laboratory students.

  15. Analysis of Chromothripsis by Combined FISH and Microarray Analysis.

    Science.gov (United States)

    MacKinnon, Ruth N

    2018-01-01

    Fluorescence in situ hybridization (FISH) to metaphase chromosomes, in conjunction with SNP array, array CGH, or whole genome sequencing, can help determine the organization of abnormal genomes after chromothripsis and other types of complex genome rearrangement. DNA microarrays can identify the changes in copy number, but they do not give information on the organization of the abnormal chromosomes, balanced rearrangements, or abnormalities of the centromeres and other regions comprised of highly repetitive DNA. Many of these details can be determined by the strategic use of metaphase FISH. FISH is a single-cell technique, so it can identify low-frequency chromosome abnormalities, and it can determine which chromosome abnormalities occur in the same or different clonal populations. These are important considerations in cancer. Metaphase chromosomes are intact, so information about abnormalities of the chromosome homologues is preserved. Here we describe strategies for working out the organization of highly rearranged genomes by combining SNP array data with various metaphase FISH methods. This approach can also be used to address some of the uncertainties arising from whole genome or mate-pair sequencing data.

  16. Impact of fishing and stocking practices on Coregonid diversity

    Science.gov (United States)

    Anneville, Orlane; Lasne, Emilien; Guillard, Jean; Eckmann, Reiner; Stockwell, Jason D.; Gillet, Christian; Yule, Daniel

    2015-01-01

    Fish species diversity can be lost through interacting stressors including habitat loss, stocking and overfishing. Although a multitude of stressors have played a role in the global decline of coregonid (Coregonus spp.) diversity, a number of contemporary studies have identified habitat loss stemming from eutrophication as the primary cause. Unfortunately, reconstructing the role of fishing and stocking practices can be difficult, because these records are incomplete or appear only in hard-to-access historic grey literature. Based on an illustrative set of historic and contemporary studies, we describe how fisheries management practices may have contributed to coregonid diversity loss in European and North American lakes. We provide case studies examining how fishing and stocking may reduce coregonid diversity through demographic decline and introgressive hybridization. In some lakes, fisheries management practices may have led to a loss of coregonid diversity well before issues with habitat degradation manifested. Our review suggests that fish conservation policies could beneficially consider the relative importance of all stressors, including management practices, as potential drivers of diversity loss.

  17. Lower hybrid current drive: an overview of simulation models, benchmarking with experiment, and predictions for future devices

    International Nuclear Information System (INIS)

    Bonoli, P.T.; Barbato, E.; Imbeaux, F.

    2003-01-01

    This paper reviews the status of lower hybrid current drive (LHCD) simulation and modeling. We first discuss modules used for wave propagation, absorption, and current drive with particular emphasis placed on comparing exact numerical solutions of the Fokker Planck equation in 2-dimension with solution methods that employ 1-dimensional and adjoint approaches. We also survey model predictions for LHCD in past and present experiments showing detailed comparisons between simulated and observed current drive efficiencies and hard X-ray profiles. Finally we discuss several model predictions for lower hybrid current profile control in proposed next step reactor options. (authors)

  18. Contribution of fluorescence in situ hybridization to biological dosimetry

    International Nuclear Information System (INIS)

    Sorokine-Durm, I.; Roy, L.; Durand, V.; Voisin, P.

    1995-01-01

    Fluorescence in situ hybridization with composite whole chromosome specific DNA probes for human chromosomes 2, 4 and 12 an α-satellite centromeric DNA probe labelled with biotin were used to measure symmetrical and terminal translocations (dose rate 0.5 Gy/min) and dicentrics (0.1 Gy/min) induced in vitro by 60 Co γ-irradiation (0-5 Gy). The suitability of fluorescence in situ hybridization (F.I.S.H.) technique for dicentrics detection is compared with the conventional technique. Dose-response curves for γ-rays ( 60 Co) for two dose rates are shown (dicentrics and translocations). (authors). 10 refs., 2 figs

  19. [Effects of fish on field resource utilization and rice growth in rice-fish coculture].

    Science.gov (United States)

    Zhang, Jian; Hu, Liang Liang; Ren, Wei Zheng; Guo, Liang; Wu, Min Fang; Tang, Jian Jun; Chen, Xin

    2017-01-01

    Rice field can provide habitat for fish and other aquatic animals. Rice-fish coculture can increase rice yield and simultaneously reduce the use of chemicals through reducing rice pest occurrence and nutrient complementary use. However, how fish uses food sources (e.g. phytoplankton, weeds, duckweed, macro-algal and snail) from rice field, and whether the nutrients releasing from those food sources due to fish transforming can improve rice growth are still unknown. Here, we conducted two field experiments to address these questions. One was to investigate the pattern of fish activity in the field using the method of video recording. The other was to examine the utilization of field resources by fish using stable isotope technology. Rice growth and rice yield were also exa-mined. Results showed that fish tended to be more active and significantly expanded the activity range in the rice-fish coculture compared to fish monoculture (fish not living together with rice plants). The contributions of 3 potential aquatic organisms (duckweed, phytoplankton and snail) to fish dietary were 22.7%, 34.8% and 30.0% respectively under rice-fish coculture without feed. Under the treatment with feed, however, the contributions of these 3 aquatic organisms to the fish die-tary were 8.9%, 5.9% and 1.6% respectively. The feed contribution was 71.0%. Rice-fish coculture significantly increased the nitrogen concentration in rice leaves, prolonged tillering stage by 10-12 days and increased rice spike rate and yield. The results suggested that raising fish in paddy field may transform the nutrients contained in field resources to bioavailable for rice plants through fish feeding activity, which can improve rice growth and rice yield.

  20. speciation through asexuality in fish: postzygotic reproductive isolation may be completed in spite of fertility of hybrids

    Directory of Open Access Journals (Sweden)

    Karel Janko

    2015-12-01

    Altogether, it appears that initiation of hybrid asexuality and the completion of speciation process through formation of postRIMs are interconnected phenomena. Both processes are linked to the genetic divergence of hybridizing taxa: initially, hybridization between little diverged species leads to recombinant and fertile hybrids allowing intensive gene flow. As the hybridizing taxa continue to diverge, clonally reproducing hybrid females and sterile males become dominant and the gene flow ceases. The speciation may therefore be completed through asexuality of hybrids The work was supported by grant no. 13-12580S provided by the Czech Science Foundation (www.gacr.cz. Further support was provided by the Academy of Sciences of the Czech Republic (www.cas.cz by the grant no. RVO 67985904

  1. Comparison of the chromosome maps around a resistance hot spot on chromosome 5 of potato and tomato using BAC-FISH painting.

    Science.gov (United States)

    Achenbach, Ute C; Tang, Xiaomin; Ballvora, Agim; de Jong, Hans; Gebhardt, Christiane

    2010-02-01

    Potato chromosome 5 harbours numerous genes for important qualitative and quantitative traits, such as resistance to the root cyst nematode Globodera pallida and the late blight fungus, Phytophthora infestans. The genes make up part of a "hot spot" for resistances to various pathogens covering a genetic map length of 3 cM between markers GP21 and GP179. We established the physical size and position of this region on chromosome 5 in potato and tomato using fluorescence in situ hybridization (FISH) on pachytene chromosomes. Five potato bacterial artificial chromosome (BAC) clones with the genetically anchored markers GP21, R1-contig (proximal end), CosA, GP179, and StPto were selected, labeled with different fluorophores, and hybridized in a five-colour FISH experiment. Our results showed the location of the BAC clones in the middle of the long arm of chromosome 5 in both potato and tomato. Based on chromosome measurements, we estimate the physical size of the GP21-GP179 interval at 0.85 Mb and 1.2 Mb in potato and tomato, respectively. The GP21-GP179 interval is part of a genome segment known to have inverted map positions between potato and tomato.

  2. Transit experience with hydrogen fueled hybrid electric buses

    Energy Technology Data Exchange (ETDEWEB)

    Scott, P.B.; Mazaika, D.M. [ISE Corp., Poway, CA (United States)

    2006-07-01

    Mass transit buses are ideal candidates for hydrogen implementation due to their capability of carrying 30 to 60 kg of hydrogen. ISE Corporation is a supplier of hydrogen fueled buses, including the first hybrid electric fuel cell bus which was commercialized in 2002, the hybrid electric fuel cell bus, and the hybrid hydrogen internal combustion engine (HHICE) bus which was commercialized in 2004. The configuration of a HHICE bus was illustrated with reference to its engine, control system, energy storage, generator, drive motor, inverter and accessories. Although these vehicles are expensive, the cost is amortized over a large base of hours used and passengers carried. The buses are operated primarily in urban areas where quiet and clean operation is needed the most. ISE has established a joint venture with Thor industries to develop a series of fuel cell buses equipped with a 60 kW PEM fuel cell. A schematic illustrating the energy flow in HHICE bus was also presented. It was shown that regenerative braking recovers the energy of motion. When using regenerative braking, most of the braking energy is saved in the battery. ISE drive systems convert 30 per cent or more of the bus energy to electrical energy to be used in later acceleration. Reduced fuel consumption also reduces the vehicle emissions. Testing of HHICE buses in both summer and winter operating conditions have shown that the range needs to be improved along with engine component reliability and durability. Fuel supply is also a major issue. A comparison with a fuel cell hybrid system was also presented. In the United States, more than 100,000 miles have been logged for the use of hydrogen hybrid buses, fuel cell buses and HHICE buses. The HHICE bus offers low capital cost, familiar technologies, but some NOx. CAT absorber technology offers the possibility of near zero emission capability. The fuel cell bus was found to be more fuel efficient, and can travel nearly twice as far per unit energy as

  3. Divergent mating preferences and nuptial coloration in sibling species of cichlid fish

    NARCIS (Netherlands)

    Sluijs, Inke van der

    2008-01-01

    Mate choice by female cichlid fish from Lake Victoria plays an important role in speciation and the maintenance of species. Females are expected to select against males that are intermediate in their phenotype during the process of speciation driven by sexual selection. To test this, we hybridized

  4. Genome constitution and evolution in Lolium X Festuca hybrid cultivars (Festulolium)

    Czech Academy of Sciences Publication Activity Database

    Kopecký, David; Loureiro, J.; Zwierzykowski, Z.; Ghesquière, M.; Doležel, Jaroslav

    2006-01-01

    Roč. 113, - (2006), s. 731-742 ISSN 0040-5752 R&D Projects: GA AV ČR IBS5038104 Institutional research plan: CEZ:AV0Z50380511 Keywords : Festulolium hybrids * GISH * FISH Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.715, year: 2006

  5. Use of number by fish.

    Directory of Open Access Journals (Sweden)

    Christian Agrillo

    Full Text Available BACKGROUND: Research on human infants, mammals, birds and fish has demonstrated that rudimentary numerical abilities pre-date the evolution of human language. Yet there is controversy as to whether animals represent numbers mentally or rather base their judgments on non-numerical perceptual variables that co-vary with numerosity. To date, mental representation of number has been convincingly documented only for a few mammals. METHODOLOGY/PRINCIPAL FINDINGS: Here we used a training procedure to investigate whether mosquitofish could learn to discriminate between two and three objects even when denied access to non-numerical information. In the first experiment, fish were trained to discriminate between two sets of geometric figures. These varied in shape, size, brightness and distance, but no control for non-numerical variables was made. Subjects were then re-tested while controlling for one non-numerical variable at a time. Total luminance of the stimuli and the sum of perimeter of figures appeared irrelevant, but performance dropped to chance level when stimuli were matched for the cumulative surface area or for the overall space occupied by the arrays, indicating that these latter cues had been spontaneously used by the fish during the learning process. In a second experiment, where the task consisted of discriminating 2 vs 3 elements with all non-numerical variables simultaneously controlled for, all subjects proved able to learn the discrimination, and interestingly they did not make more errors than the fish in Experiment 1 that could access non-numerical information in order to accomplish the task. CONCLUSIONS/SIGNIFICANCE: Mosquitofish can learn to discriminate small quantities, even when non-numerical indicators of quantity are unavailable, hence providing the first evidence that fish, like primates, can use numbers. As in humans and non-human primates, genuine counting appears to be a 'last resort' strategy in fish, when no other

  6. Three sympatric karyomorphs in the fish Astyanax fasciatus (Teleostei, Characidae do not seem to hybridize in natural populations

    Directory of Open Access Journals (Sweden)

    Maressa Ferreira-Neto

    2012-01-01

    Full Text Available Ninety individuals of the characid fish Astyanax fasciatus (Cuvier, 1819 were collected at Água da Madalena stream (Botucatu, São Paulo, Brazil and analyzed for diploid chromosome number 2n and karyotype composition as well as for the chromosomal location of the 5S and 18S ribosomal DNA (rDNA. Whereas no chromosome differences were associated with sex, three different karyomorphs with diploid chromosome numbers 2n=46, 2n=48 and 2n=50 were found. No intermediate 2n numbers were discovered. The 2n=50 karyomorph showed some differences in 18S rDNA location compared to the two other karyomorphs. Finally, all specimens with the 2n=46 karyomorph showed the presence of a partly heterochromatic macro supernumerary chromosome, which was absent in all individuals with the two other karyomorphs. All these results suggest that indviduals of the three different karyomorphs are not likely to hybridize in the examined populations. Our findings strongly suggest the presence of three separate species (sensu biological species concept easily diagnosed on the basis of differences in the diploid chromosome numbers and other chromosomal markers.

  7. Evaluation of upper urinary tract tumors by FISH in Chinese patients.

    Science.gov (United States)

    Shan, Zhengfei; Wu, Peng; Zheng, Shaobin; Tan, Wanlong; Zhou, Haikuan; Zuo, Yi; Qi, Huan; Zhang, Peng; Peng, Hongmei; Wang, Yanfen

    2010-12-01

    Upper urinary tract tumor (UUTT) usually presents a high grade and stage, and recurs frequently. The aim of this study was to evaluate the utility of a fluorescence in situ hybridization (FISH) assay on chromosomes 3, 7, 9, and 17 as a reliable and noninvasive method for the diagnosis of Chinese patients with UUTT. Urine specimens from 50 patients with UUTT and 25 donors without evidence of urothelial tumors were analyzed by cytology and FISH. Voided urine samples from 20 normal individuals were used to establish the cut-off values for FISH assay. The McNemar test was applied for sensitivity and specificity. The overall sensitivity of FISH was statistically significantly greater than that of cytology (84.0 vs. 40.0%, P = 0.000). The overall specificities of FISH and urine cytology were all 96.0% (P = 1.000). Polysomy in chromosomes 3, 7, and 17 were 38, 42, and 30%, respectively. Heterozygous and homozygous loss of the p16 locus was found in 36 and 32%, respectively. FISH analysis performed on cells collected from voided urine is feasible, and FISH could prove to be a reliable and less invasive ancillary test and improve the sensitivity of urine cytology in the diagnosis of UUTT. Copyright © 2010 Elsevier Inc. All rights reserved.

  8. Using copper sulfate to control egg fungus at Keo Fish Farm

    Science.gov (United States)

    Keo Fish Farm is the biggest producer of hybrid striped bass fry in the world. The hatchery manager asked about treatments to control fungus on eggs which occurred fairly often. Our lab has been working on gaining FDA-approval to use copper sulfate to control fungus on catfish eggs, so we were con...

  9. Optimization of three FISH procedures for in situ detection of anaerobic ammonium oxidizing bacteria in biological wastewater treatment.

    Science.gov (United States)

    Pavlekovic, Marko; Schmid, Markus C; Schmider-Poignee, Nadja; Spring, Stefan; Pilhofer, Martin; Gaul, Tobias; Fiandaca, Mark; Löffler, Frank E; Jetten, Mike; Schleifer, K-H; Lee, Natuschka M

    2009-08-01

    Fluorescence in situ hybridization (FISH) using fluorochrome-labeled DNA oligonucleotide probes has been successfully applied for in situ detection of anaerobic ammonium oxidizing (anammox) bacteria. However, application of the standard FISH protocols to visualize anammox bacteria in biofilms from a laboratory-scale wastewater reactor produced only weak signals. Increased signal intensity was achieved either by modifying the standard FISH protocol, using peptide nucleic acid probes (PNA FISH), or applying horse radish peroxidase- (HRP-) labeled probes and subsequent catalyzed reporter deposition (CARD-FISH). A comparative analysis using anammox biofilm samples and suspended anammox biomass from different laboratory wastewater bioreactors revealed that the modified standard FISH protocol and the PNA FISH probes produced equally strong fluorescence signals on suspended biomass, but only weak signals were obtained with the biofilm samples. The probe signal intensities in the biofilm samples could be enhanced by enzymatic pre-treatment of fixed cells, and by increasing the hybridization time of the PNA FISH protocol. CARD-FISH always produced up to four-fold stronger fluorescent signals but unspecific fluorescence signals, likely caused by endogenous peroxidases as reported in several previous studies, compromised the results. Interference of the development of fluorescence intensity with endogenous peroxidases was also observed in cells of aerobic ammonium oxidizers like Nitrosomonas europea, and sulfate-reducers like Desulfobacter postgatei. Interestingly, no interference was observed with other peroxidase-positive microorganisms, suggesting that CARD-FISH is not only compromised by the mere presence of peroxidases. Pre-treatment of cells to inactivate peroxidase with HCl or autoclavation/pasteurization failed to inactive peroxidases, but H(2)O(2) significantly reduced endogenous peroxidase activity. However, for optimal inactivation, different H(2)O(2

  10. Implementation and importance of fluorescence in situ hybridization (fish) in paraffin tissues for categorization of B-cell lymphoma unclassifiable, with features intermediate between Burkitt lymphoma and diffuse large B-cell lymphoma

    International Nuclear Information System (INIS)

    Carvajal Cuenca, Alejandra

    2011-01-01

    The diagnostic criteria have been defined based on the tools that the country has acquired and international guidelines for pure entities: the LB, LDCGB, and the new entity of B lymphoma unclassifiable with features intermediate LDCGB and LB. The fluorescence in situ hybridization for the translocation (8;14) has been implemented in paraffin tissues for proper categorization. A total of 21 cases have been studied: the characteristics of patients, morphology, immunohistochemistry and the presence or absence of the translocation (8;14). Twelve of the cases have been classified as B-cell lymphoma unclassifiable with features intermediate between LDCGB and LB. Furthermore, nine of the cases were classified in LB. Fluorescence in situ hybridization (FISH) has been negative in 5 of the 21 cases. The diagnosis of lymphoma with features bordering between the LB and the LDCGB has been imperative for the survival of the patient and the corresponding treatment [es

  11. TECHNOLOGY CHARACTERIZATION OF THE FISH FARM OF THE MUNICIPALITY OF CAPITÃO POÇO, PARÁ, BRAZIL

    Directory of Open Access Journals (Sweden)

    Tiago Pereira Brito

    2017-07-01

    Full Text Available The present study describes the technological aspects of fish farming developed on the Capitão Poço in northeastern state of Pará. The data were collected through 17 semi-structured interviews in aquaculture properties in the region.The farmer was legally responsible for most property. The property's operating time ranged from 3 to 20 years and the time of experience in aquaculture activity from 10 months to 20 years (mean 5.4 ± 2.9 years. Aquaculture represented a secondary activity is for subsistence or to supplement the family income. The properties had a total area ranging from 0.5 to 750 hectares (average 124.9 ± 150.0 ha, with water area between 0.00165 and 2.16 ha (average 0.4944 ± 0.5694 ha. The semi-intensive production system was predominant (88.2%, however, the intensive system keeper tanks was also recorded (11.8%, and recorded both monocultures (70.6% as policultives (29, 4%, with cultive of Oreochromis niloticus, Colossoma macropomum, Leporinus friderici and two species hybrid. The management with regard to the analysis of water quality of farming, food supply and evaluation of growth performance, although carried out were not satisfactory, requiring greater technical training to producers. Keywords Amazon; aquaculture; farmer fish; fish; cultivation system.

  12. Lactobacillus genus identification isolated from gastrointestinal tract of chickens after bee products application using FISH and RTQ PCR methods

    Directory of Open Access Journals (Sweden)

    Miroslava Kačániová

    2013-05-01

    Full Text Available The general objective of this study was to examine the effect of bee products on the lactobacilli colonization of chickens. Bee products were administered to both feed mixtures in various amounts in addition to the control group. First experimental group was with propolis in feed mixture with the addition of 400 mg propolis per 1 kg of compound and second group was with pollen in feed mixture with the addition of 450 mg pollen per 1 kg of compound. In this experiment, quantitative counts of lactobacilli in ceca of 49-day-old chicken (Ross 308 using classical and FISH method were investigated. Counts of lactobacilli on MRS agar were monitored. To check the reliability of traditional methods of cultivation samples were evaluated by fluorescence in situ hybridization (FISH. Lactobacillus cells, isolated from gastrointestinal tract, were detected after hybridization of fluorescently labeled probe with bacterial cells. Counts of CFU of lactobacilli were compared in experimental and control treatments, respectively. The lowest count was detected in the control experimental group. The highest count was detected in the third experimental group where was 450 mg of pollen added to 1 kg of feed mixture. Using Real-time PCR method, we identified the species range of the genera Lactobacillus in the intestinal tract of broiler. Detected species from the genus Lactobacillus were L. crispatus, L. salivarius and L. acidophilus.

  13. Population structure and adaptation in fishes

    DEFF Research Database (Denmark)

    Limborg, Morten

    Marine fishes represent a valuable resource for the global economy and food consumption. Accordingly, many species experience high levels of exploitation necessitating effective management plans. However, long term sustainability may be jeopardized from insufficient knowledge about intra-specific......Marine fishes represent a valuable resource for the global economy and food consumption. Accordingly, many species experience high levels of exploitation necessitating effective management plans. However, long term sustainability may be jeopardized from insufficient knowledge about intra......-specific population structure and adaptive divergence. The large population sizes and high migration rates common to most marine fishes impede the differentiating effect of genetic drift, having led to expectations of no population structure and that the occurrence of local adaptation should be rare in these species....... Comprehensive genetic analyses on the small pelagic fish European sprat (Sprattus sprattus) revealed significant population structure throughout its distribution with an overall pattern of reduced connectivity across environmental transition zones. Population structure reflected both historical separations over...

  14. A Novel Artificial Fish Swarm Algorithm for Recalibration of Fiber Optic Gyroscope Error Parameters

    Directory of Open Access Journals (Sweden)

    Yanbin Gao

    2015-05-01

    Full Text Available The artificial fish swarm algorithm (AFSA is one of the state-of-the-art swarm intelligent techniques, which is widely utilized for optimization purposes. Fiber optic gyroscope (FOG error parameters such as scale factors, biases and misalignment errors are relatively unstable, especially with the environmental disturbances and the aging of fiber coils. These uncalibrated error parameters are the main reasons that the precision of FOG-based strapdown inertial navigation system (SINS degraded. This research is mainly on the application of a novel artificial fish swarm algorithm (NAFSA on FOG error coefficients recalibration/identification. First, the NAFSA avoided the demerits (e.g., lack of using artificial fishes’ pervious experiences, lack of existing balance between exploration and exploitation, and high computational cost of the standard AFSA during the optimization process. To solve these weak points, functional behaviors and the overall procedures of AFSA have been improved with some parameters eliminated and several supplementary parameters added. Second, a hybrid FOG error coefficients recalibration algorithm has been proposed based on NAFSA and Monte Carlo simulation (MCS approaches. This combination leads to maximum utilization of the involved approaches for FOG error coefficients recalibration. After that, the NAFSA is verified with simulation and experiments and its priorities are compared with that of the conventional calibration method and optimal AFSA. Results demonstrate high efficiency of the NAFSA on FOG error coefficients recalibration.

  15. Effects of fire on fish populations: Landscape perspectives on persistance of native fishes and nonnative fish invasions

    Science.gov (United States)

    Dunham, J.B.; Young, M.; Gresswell, Robert E.; Rieman, B.

    2003-01-01

    Our limited understanding of the short and long-term effects of fire on fish contributes to considerable uncertainty in assessments of the risks and benefits of fire management alternatives. A primary concern among the many potential effects of fire is the effects of fire and fire management on persistence of native fish populations. Limited evidence suggests vulnerability of fish to fire is contingent upon the quality of affected habitats, the amount and distribution of habitat (habitat fragmentation), and habitat specificity of the species in question. Species with narrow habitat requirements in highly degraded and fragmented systems are likely to be most vulnerable to fire and fire-related disturbance. In addition to effects of fire on native fish, there are growing concerns about the effects of fire on nonnative fish invasions. The role of fire in facilitating invasions by nonnative fishes is unknown, but experience with other species suggests some forms of disturbance associated with fire may facilitate invasion. Management efforts to promote persistence of fishes in fire-prone landscapes can take the form of four basic alternatives: (1) pre-fire management; (2) post-fire management; (3) managing fire itself (e.g. fire fighting); and (4) monitoring and adaptive management. Among these alternatives, pre-fire management is likely to be most effective. Effective pre-fire management activities will address factors that may render fish populations more vulnerable to the effects of fire (e.g. habitat degradation, fragmentation, and nonnative species). Post-fire management is also potentially important, but suffers from being a reactive approach that may not address threats in time to avert them. Managing fire itself can be important in some contexts, but negative consequences for fish populations are possible (e.g. toxicity of fire fighting chemicals to fish). Monitoring and adaptive management can provide important new information for evaluating alternatives, but

  16. Snapshots of past fish faunas

    DEFF Research Database (Denmark)

    Enghoff, Inge Bødker; Ediger, Vedat

    2016-01-01

    Analyses of fish remains from sediment cores make it possible to detect not only commonly caught fish from prehistoric times, but also species without any economic importance, but with high value of paleaoecological reconstructions. In this study, fish bones from sediment cores reaching several...... thousand years back and taken in the Baltic and Black Seas were analysed. All fish remains dealt with postdate the last glaciations and are from the last marine/brackish stages of both seas. In the Baltic cores, 13+ species were found, the most abundant ones being sand-eel and clupeids (herring and sprat...... before industrial fishing for them began. Clupeids, in the Baltic samples also sand-eel, dominate the materials. Both contain species that would hardly be expected on archaeological sites. Experience from this study leads to methodological recommendations regarding dating of material from sediment cores...

  17. Molecular cytogenetic analysis of human blastocysts andcytotrophoblasts by multi-color FISH and Spectra Imaging analyses

    Energy Technology Data Exchange (ETDEWEB)

    Weier, Jingly F.; Ferlatte, Christy; Baumgartner, Adolf; Jung,Christine J.; Nguyen, Ha-Nam; Chu, Lisa W.; Pedersen, Roger A.; Fisher,Susan J.; Weier, Heinz-Ulrich G.

    2006-02-08

    Numerical chromosome aberrations in gametes typically lead to failed fertilization, spontaneous abortion or a chromosomally abnormal fetus. By means of preimplantation genetic diagnosis (PGD), we now can screen human embryos in vitro for aneuploidy before transferring the embryos to the uterus. PGD allows us to select unaffected embryos for transfer and increases the implantation rate in in vitro fertilization programs. Molecular cytogenetic analyses using multi-color fluorescence in situ hybridization (FISH) of blastomeres have become the major tool for preimplantation genetic screening of aneuploidy. However, current FISH technology can test for only a small number of chromosome abnormalities and hitherto failed to increase the pregnancy rates as expected. We are in the process of developing technologies to score all 24 chromosomes in single cells within a 3 day time limit, which we believe is vital to the clinical setting. Also, human placental cytotrophoblasts (CTBs) at the fetal-maternal interface acquire aneuploidies as they differentiate to an invasive phenotype. About 20-50% of invasive CTB cells from uncomplicated pregnancies were found aneuploidy, suggesting that the acquisition of aneuploidy is an important component of normal placentation, perhaps limiting the proliferative and invasive potential of CTBs. Since most invasive CTBs are interphase cells and possess extreme heterogeneity, we applied multi-color FISH and repeated hybridizations to investigate individual CTBs. In summary, this study demonstrates the strength of Spectral Imaging analysis and repeated hybridizations, which provides a basis for full karyotype analysis of single interphase cells.

  18. Hybrid Ground-Source Heat Pump Installations: Experiences, Improvements, and Tools

    Energy Technology Data Exchange (ETDEWEB)

    Scott Hackel; Amanda Pertzborn

    2011-06-30

    One innovation to ground-source heat pump (GSHP, or GHP) systems is the hybrid GSHP (HyGSHP) system, which can dramatically decrease the first cost of GSHP systems by using conventional technology (such as a cooling tower or a boiler) to meet a portion of the peak heating or cooling load. This work uses three case studies (two cooling-dominated, one heating-dominated) to demonstrate the performance of the hybrid approach. Three buildings were studied for a year; the measured data was used to validate models of each system. The models were used to analyze further improvements to the hybrid approach, and establish that this approach has positive impacts, both economically and environmentally. Lessons learned by those who design and operate the systems are also documented, including discussions of equipment sizing, pump operation, and cooling tower control. Finally, the measured data sets and models that were created during this work are described; these materials have been made freely available for further study of hybrid systems.

  19. Marine soundscape shaped by fishing activity.

    Science.gov (United States)

    Coquereau, Laura; Lossent, Julie; Grall, Jacques; Chauvaud, Laurent

    2017-01-01

    Marine communities face anthropogenic pressures that degrade ecosystems. Because underwater soundscapes carry information about habitat quality, we explored whether destructive impacts of fishing could be evaluated via the soundscape. Maerl beds are recognized as biodiversity hotspots and they experience major worldwide degradation owing to fishing. We collected field acoustic recordings in maerl beds exposed to different fishing practices. We found that unfished maerl beds were threefold louder and exhibited sound frequencies more diversified than those recorded in fished maerl beds. Analyses of associated fauna samples indicated that snapping shrimps provided a major contribution to the maerl bed soundscape. Moreover, sea urchins and squat lobsters most likely contributed to differences between the soundscapes of unfished and fished maerl beds. Our results supported the idea that the soundscape can provide valuable information on maerl bed ecosystem health related to fishing activity.

  20. THE CLASSIC WAY OF FISH PROCESSING

    Directory of Open Access Journals (Sweden)

    Đurica Kalember

    1998-04-01

    Full Text Available Today's population faces great difficulties in fish marketing, although it is very valuable food. The classic supply with fresh fish has little influence on its consumption, which is not remarkable anyhow. Therefore one shulud be reminded on the classic, almost forgotten, ways of fish processing that can substantially increase fish assortment and improve its distribution. After cleaning and cutting the fish (primary procedures in its processing, comes salting, after which the salted fish can become an end-product or it can be one of many semi-products in the fish production chain. The most common methods of fish salting are dry-salting, dry-wet-salting (Greek-Dalmatian and wet-salting (pickling. The aim of fish drying is its dehydratation. Our country has the experience of traditional drying, sun-drying and natural drying of fish. Each of these has its own special qualities, depending on the fish species and the drying temperature. Smoked fish gets a very distinctive and spicy aroma and a specific colour. There are two kinds of smoking - cold and warm - based on the smoke derived from burning some special trees or, lately, from smoke preparations. Marinades are old procedures of fish processing in acetic acid and specific spices which can be prepared cold, fried or cooked. Fish-roe of some specific fish species has a special value and is considered a delicacy. The most precious black caviar is derived from the sturgeon roe and some of its related species.

  1. Single molecule localization imaging of telomeres and centromeres using fluorescence in situ hybridization and semiconductor quantum dots.

    Science.gov (United States)

    Wang, Le; Zong, Shenfei; Wang, Zhuyuan; Lu, Ju; Chen, Chen; Zhang, Ruohu; Cui, Yiping

    2018-07-13

    Single molecule localization microscopy (SMLM) is a powerful tool for imaging biological targets at the nanoscale. In this report, we present SMLM imaging of telomeres and centromeres using fluorescence in situ hybridization (FISH). The FISH probes were fabricated by decorating CdSSe/ZnS quantum dots (QDs) with telomere or centromere complementary DNA strands. SMLM imaging experiments using commercially available peptide nucleic acid (PNA) probes labeled with organic fluorophores were also conducted to demonstrate the advantages of using QDs FISH probes. Compared with the PNA probes, the QDs probes have the following merits. First, the fluorescence blinking of QDs can be realized in aqueous solution or PBS buffer without thiol, which is a key buffer component for organic fluorophores' blinking. Second, fluorescence blinking of the QDs probe needs only one excitation light (i.e. 405 nm). While fluorescence blinking of the organic fluorophores usually requires two illumination lights, that is, the activation light (i.e. 405 nm) and the imaging light. Third, the high quantum yield, multiple switching times and a good optical stability make the QDs more suitable for long-term imaging. The localization precision achieved in telomeres and centromeres imaging experiments is about 30 nm, which is far beyond the diffraction limit. SMLM has enabled new insights into telomeres or centromeres on the molecular level, and it is even possible to determine the length of telomere and become a potential technique for telomere-related investigation.

  2. Impact of a widely cultivated tree (moringa oleifera) on the health of commercially important hybrid catfish

    International Nuclear Information System (INIS)

    Adamu, K.M.; Ahmed, I.M.

    2014-01-01

    Plantations of the tree Moringa oleifera often lead to increase levels of leaf dust in nearby freshwater environments, and there is concern that, this could have a negative impact on catfish, which are important for aquaculture. Therefore, this study, determined the biochemical alterations in serum, liver and kidney of hybrid catfish (Clarias gariepinus ( ) Heterobranchus bidorsalis ( ) exposed to sub-lethal concentrations of M. oleifera leaf dust in a static renewable bioassay system during a medium term exposure period. The fish (mean length, 16.33 cm, mean weight, 9.90 g) were exposed to 0.16, 0.12, 0.08, 0.04 and 0.00 mg/L concentrations of the plant leaf dust in triplicate exposure. After 21-days of exposure period, the fish were sacrificed for the biochemical parameters: glucose, total protein, albumin, globulin, cholesterol, triglyceride, g-glutamyltransferase, alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase. Fish exposed to leaf dust showed significant differences (P 0.05), in the liver and kidney, respectively, in the test fish. Other parameters were not significantly different (P>0.05) in their respective tissue-organs. Ipso-facto, the alteration in biochemical parameters of hybrid catfish exposed to M. oleifera leaf dust was concentration dependent with 0.16 mg/L showing the highest negative alterations thus fish exposed to concentrations above 0.16 mg/L for longer durations may suffer impaired health effect. (author)

  3. Impacts of different rice-fish-prawn culture systems on yield of rice, fish and prawn and limnological conditions

    OpenAIRE

    Nahar, Ashfaqun

    2010-01-01

    An experiment was conducted to determine the impact of fish and prawn culture on some physicochemical parameters of water, weeds, benthos, and rice yield under simultaneous method for a period of 5 months from July to November 2007. The experiment was comprised of five individual treatments having three replicates for each. The treatments were: rice combined with fish and regular urea fertilization (treatment І, T1), rice combined with prawn and regular urea fertilization (treatment ІI, T2), ...

  4. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    Energy Technology Data Exchange (ETDEWEB)

    Rosa, F.E.; Santos, R.M. [Departamento de Patologia, Hospital das Clínicas, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil); Rogatto, S.R. [Departamento de Urologia, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil); Hospital A.C. Camargo, CIPE, São Paulo, SP (Brazil); Domingues, M.A.C. [Departamento de Patologia, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil)

    2013-03-19

    Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

  5. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    Science.gov (United States)

    Rosa, F.E.; Santos, R.M.; Rogatto, S.R.; Domingues, M.A.C.

    2013-01-01

    Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer. PMID:23558859

  6. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    International Nuclear Information System (INIS)

    Rosa, F.E.; Santos, R.M.; Rogatto, S.R.; Domingues, M.A.C.

    2013-01-01

    Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer

  7. Evaluation of Myc Gene Amplification in Prostate Cancer Using a Dual Color Chromogenic in-Situ Hybridization (Dual CISH Assay

    Directory of Open Access Journals (Sweden)

    Daniel Lerda

    2013-04-01

    Full Text Available Objetive: The overall purpose of the study was to demonstrate applicability of the Dako dual-color chromogenic in situ hybridization (CISH assay (DAKO Denmark, Glostrup with respect to fluorescence in situ hybridization (FISH probes MYC-C. Methods: MYC gene amplification by FISH and Dako dual-color CISH Results: The study showed that the dual-color CISH assay can convert Texas red and fluorescein isothiocyanate (FITC signals into chromogenic signals. The dual –color CISH assay was performed on 40 cases of prostate cancer. Amplification was identified in 12 of 40 (30% tumors. No amplification was seen in 28 of 40 (70% tumors. FISH data were available in total of 40 tumors. All tumors showed concordant results between dual-color CISH and FISH for classifying a tumor as MYC amplified or not amplified. Conclusions: We conclude that dual-color Dako CISH assay is an accurate method for determining MYC gene amplification with added advantages that make it a more practically useful method. [J Interdiscipl Histopathol 2013; 1(2.000: 81-84

  8. FISH analysis for diagnostic evaluation of challenging melanocytic lesions.

    Science.gov (United States)

    Zimmermann, A K; Hirschmann, A; Pfeiffer, D; Paredes, B E; Diebold, J

    2010-09-01

    The differential diagnosis of malignant melanomas and atypical melanocytic nevi is still a diagnostic challenge. The currently accepted morphologic criteria show substantial interobserver variability, likewise immunohistochemical studies are often not able to discriminate these lesions reliably. Techniques that support diagnostic accuracy are of the greatest importance considering the growing incidence of malignant melanomas and their increase in younger patients. In this study we analyzed the feasibility of fluorescence in situ hybridization (FISH) analysis for the discrimination of malignant and benign melanocytic tumors. A panel of DNA probes was used to detect chromosomal aberrations of chromosomes 6 and 11. On a series of 5 clearly malignant and benign melanocytic tumors we confirmed the applicability of the test. Then we focused on examination of ambiguous melanocytic lesions, where atypical cells are often difficult to relocalize in the 4',6-Diamidino-2-phenylindol (DAPI)-fluorescence stain. FISH analyses were conducted on destained H&E-stained slides. By comparison of the DAPI-image with photos taken from the H&E stain, unambiguous assignment of the FISH results to the conspicuous groups of cells was possible. The results of FISH analysis were consistent with the conventional diagnosis in 11 of 14 small ambiguous lesions. Of the remaining 3 cases, 2 showed FISH-results close to the cut-off level. Comparison of FISH results on thin and thick sections revealed that the cut-off values have to be adapted for 2 microm destained sections. In conclusion, FISH analysis is a useful and applicable tool for assessment of even smallest melanocytic neoplasms, although there will remain unclear cases that cannot be solved even after additional FISH evaluation.

  9. Evaluation of gene amplification and protein expression of HER-2/neu in esophageal squamous cell carcinoma using Fluorescence in situ Hybridization (FISH) and immunohistochemistry

    International Nuclear Information System (INIS)

    Sato-Kuwabara, Yukie; Neves, José I; Fregnani, José HTG; Sallum, Rubens A; Soares, Fernando A

    2009-01-01

    Esophageal squamous cell carcinoma (ESCC) is the sixth most frequent neoplasia in Brazil. It is usually associated with a poor prognosis because it is often at an advanced stage when diagnosed and there is a high frequency of lymph node metastases. It is important to know what prognostic factors can facilitate diagnosis, optimize therapeutic decisions, and improve the survival of these patients. A member of the epidermal growth factor receptor (EGFR) family, c-erbB-2, has received much attention because of its therapeutic implications; however, few studies involving fluorescence in situ hybridization (FISH) analysis of HER-2/neu gene amplification and protein expression in ESCC have been conducted. The aim of this study was to verify the presence of HER-2/neu gene amplification using FISH, and to correlate the results with immunohistochemical expression and clinical-pathological findings. One hundred and ninety-nine ESCC cases were evaluated using the Tissue Microarray (TMA) technique. A polyclonal antibody against c-erbB-2 was used for immunohistochemistry. Analyses were based on the membrane staining pattern. The results were classified according to the Herceptest criteria (DAKO): negative (0/1+), potential positive (2+) and positive (3+). The FISH reactions were performed according to the FISH HER2 PharmDx (DAKO) protocol. In each case, 100 tumor nuclei were evaluated. Cases showing a gene/CEN17 fluorescence ratio ≥ 2 were considered positive for gene amplification. The c-erbB-2 expression was negative in 117/185 cases (63.2%) and positive in 68 (36.8%), of which 56 (30.3%) were 2+ and 12 (6.5%) were 3+. No significant associations were found among protein expression, clinicopathological data and overall survival. Among the 47 cases analyzed, 38 (80.9%) showed no gene amplification while 9 (19.1%) showed amplification, as demonstrated by FISH. Cases that were negative (0/1+) and potential positive (2+) for c-erbB-2 expression by immunohistochemistry showed no

  10. 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...... symptoms are linked to the presence of IgE-antibodies recognizing parvalbumin, the fish panallergen. This view was challenged by results from recent studies as follows. 1. Allergic reactions which are limited to single or several fish species (mono-or oligosensitisations) apply not only to single cases...... but patients with this phenotype constitute an important sub-group among fish-allergic individuals. 2. Newly identified fish allergens, enolases, aldolases, and fish gelatin, are of high relevance as the majority of the fish-allergic individuals seem to develop specific IgE against these proteins. The present...

  11. Analysis of the nitrifying bacterial community in BioCube sponge media using fluorescent in situ hybridization (FISH) and microelectrodes.

    Science.gov (United States)

    Chae, Kyu-Jung; Rameshwar, T; Jang, Am; Kim, Sung H; Kim, In S

    2008-09-01

    There is growing interest in the development of more cost-effective and retrofit technologies for the upgrade and expansion of existing wastewater treatment plants with extreme space constraints. A free-floating sponge media (BioCube) process, using a 24 L lab scale reactor, was operated to study the nitrification profiles and microbial community. The COD removal efficiencies were maintained, at an average of 95%, with the mixed liquor suspended solids (MLSS) inside the BioCube sponge media maintained at 12,688 mg/L. The nitrification removal efficiencies were between 92% and 100%, with an average value of 99%. From the results of microelectrode measurements, the ammonium ion concentration was found to rapidly decrease from the surface of the BioCube sponge media to a depth of 2mm due to chemical reactions carried out by ammonia oxidizing bacteria (AOB) species. Multi-fluorescence in situ hybridization (FISH) has been used to investigate the spatial distributions of various microbial activities within reactors. Microbial communities were targeted using different oligonucleotide probes specific to AOB and nitrite oxidizing bacteria (NOB). There were a large number of AOB populations, but these were not uniformly distributed in the biofilm compared to the NOB populations.

  12. Identification of embryonic chromosomal abnormality using FISH-based preimplantaion genetic diagnosis

    Institute of Scientific and Technical Information of China (English)

    叶英辉; 徐晨明; 金帆; 钱羽力

    2004-01-01

    Objective: Embryonic chromosomal abnormality is one of the main reasons for in vitro fertilization (IVF) failure. This study aimed at evaluating the value of Fluorescence in-situ Hybridization (FISH)-based Preimplantation Genetic Diagnosis (PGD) in screening for embryonic chromosomal abnormality to increase the successful rate of IVF. Method: Ten couples, four with high risk of chromosomal abnormality and six infertile couples, underwent FISH-based PGD during IVF procedure. At day 3, one or two blastomeres were aspirated from each embryo. Biopsied blastomeres were examined using FISH analysis to screen out embryos with chromosomal abnormalities. At day 4, embryos without detectable chromosomal abnormality were transferred to the mother bodies as in regular IVF. Results: Among 54 embryos screened using FISH-based PGD, 30 embryos were detected to have chromosomal abnormalities. The 24 healthy embryos were implanted, resulting in four clinical pregnancies, two of which led to successful normal birth of two healthy babies; one to ongoing pregnancy during the writing of this article; and one to ectopic pregnancy. Conclusion: FISH-based PGD is an effective method for detecting embryonic chromosomal abnormality, which is one of the common causes of spontaneous miscarriages and chromosomally unbalanced offsprings.

  13. Identification of embryonic chromosomal abnormality using FISH-based preimplantaion genetic diagnosis

    Institute of Scientific and Technical Information of China (English)

    叶英辉; 徐晨明; 金帆; 钱羽力

    2004-01-01

    Objective: Embryonic chromosomal abnormality is one of the main reasons for in vitro fertilization (IVF)failure. This study aimed at evaluating the value of Fluorescence in-situ Hybridization (FISH)-based Preimplantation Genetic Diagnosis (PGD) in screening for embryonic chromosomal abnormality to increase the successful rate of IVF. Method:Ten couples, four with high risk of chromosomal abnormality and six infertile couples, underwent FISH-based PGD during IVF procedure. At day 3, one or two blastomeres were aspirated from each embryo. Biopsied blastomeres were examined using FISH analysis to screen out embryos with chromosomal abnormalities. At day 4, embryos without detectable chromosomal abnormality were transferred to the mother bodies as in regular IVF. Results: Among 54 embryos screened using FISH-based PGD, 30 embryos were detected to have chromosomal abnormalities. The 24 healthy embryos were implanted,resulting in four clinical pregnancies, two of which led to successful normal birth of two healthy babies; one to ongoing pregnancy during the writing of this article; and one to ectopic pregnancy. Conclusion: FISH-based PGD is an effective method for detecting embryonic chromosomal abnormality, which is one of the common causes of spontaneous miscarriages and chromosomally unbalanced offsprings.

  14. Physiology of fish endocrine pancreas.

    Science.gov (United States)

    Plisetskaya, E M

    1989-06-01

    From the very beginning of physiological studies on the endocine pancreas, fish have been used as experimental subjects. Fish insulin was one of the first vertebrate insulins isolated and one of the first insulins whose primary and then tertiary structures were reported. Before a second pancreatic hormone, glucagon, was characterized, a physiologically active 'impurity', similar to that in mammalian insulin preparations, was found in fish insulins.Fish have become the most widely used model for studies of biosynthesis and processing of the pancreatic hormones. It seems inconceivable, therefore, that until the recent past cod and tuna insulins have been the only purified piscine islet hormones available for physiological experiments. The situation has changed remarkably during the last decade.In this review the contemporary status of physiological studies on the fish pancreas is outlined with an emphasis on the following topics: 1) contents of pancreatic peptides in plasma and in islet tissue; 2) actions of piscine pancreatic hormones in fish; 3) specific metabolic consequences of an acute insufficiency of pancreatic peptides; 4) functional interrelations among pancreatic peptides which differ from those of mammals. The pitfalls, lacunae and the perspectives of contemporary physiological studies on fish endocrine pancreas are outlined.

  15. Assessment of DNA damage and Chromosome aberration in human lymphocyte exposed to low dose radiation detected by FISH(Fluorescence In Situ Hybridization) and SCGE(Single Cell Gel Electrophoresis)

    International Nuclear Information System (INIS)

    Chung, Hai Won; Kim, Su Young; Kim, Byung Mo; Kim, Sun Jin; Ha, Sung Whan; Kim, Tae Hwan; Cho, Chul Koo

    2000-01-01

    Comparative study was performed for the assessment of DNA damage and Chromosomal aberration in human lymphocyte exposed to low dose radiation using Fluorescence In Situ Hybridization(FISH) and Single Cell Gel Electrophoresis(SCGE). Chromosomal aberrations in human lymphocyte exposed to radiation at doses of 5, 10, 30 and 50cGy were analysed with whole chromosome-specific probes by human chromosome 1, 2 and 4 according to PAINT system. FISH with chromosome-specific probe has been used to be a valid and rapid method for detection of chromosome rearrangements induced by low dose radiation. The frequencies of stable translocation per cell equivalents were 0.0116, 0.0375, 0.0407, 0.0727 and 0.0814 for 0, 5, 10, 30 and 50cGy, respectively, and those of dicentric were 0.00, 0.0125, 0.174, 0.0291 and 0.0407 respectively. Radiation induced DNA damage in human lymphocyte in a dose-dependent manner at low doses from 5cGy to 50cGy, which were analysed by single Cell Gel Electrophoresis(SCGE). From above results, FISH seemed to be useful for radiation biodosimetry by which the frequencies of stable aberrations in human lymphocyte can be observed more easily than by conventional method and SCGE also seemed to be sensitive method for detecting DNA damage by low dose radiation exposure, so that those methods will improve our technique to perform meaningful biodosimetry for radiation at low doses

  16. A new adaptive hybrid electromagnetic damper: modelling, optimization, and experiment

    International Nuclear Information System (INIS)

    Asadi, Ehsan; Ribeiro, Roberto; Behrad Khamesee, Mir; Khajepour, Amir

    2015-01-01

    This paper presents the development of a new electromagnetic hybrid damper which provides regenerative adaptive damping force for various applications. Recently, the introduction of electromagnetic technologies to the damping systems has provided researchers with new opportunities for the realization of adaptive semi-active damping systems with the added benefit of energy recovery. In this research, a hybrid electromagnetic damper is proposed. The hybrid damper is configured to operate with viscous and electromagnetic subsystems. The viscous medium provides a bias and fail-safe damping force while the electromagnetic component adds adaptability and the capacity for regeneration to the hybrid design. The electromagnetic component is modeled and analyzed using analytical (lumped equivalent magnetic circuit) and electromagnetic finite element method (FEM) (COMSOL ® software package) approaches. By implementing both modeling approaches, an optimization for the geometric aspects of the electromagnetic subsystem is obtained. Based on the proposed electromagnetic hybrid damping concept and the preliminary optimization solution, a prototype is designed and fabricated. A good agreement is observed between the experimental and FEM results for the magnetic field distribution and electromagnetic damping forces. These results validate the accuracy of the modeling approach and the preliminary optimization solution. An analytical model is also presented for viscous damping force, and is compared with experimental results The results show that the damper is able to produce damping coefficients of 1300 and 0–238 N s m −1 through the viscous and electromagnetic components, respectively. (paper)

  17. Comparative evaluation of non-informative HER-2 immunoreactions (2+) in breast carcinomas with FISH, CISH and QRT-PCR.

    Science.gov (United States)

    Kostopoulou, Evanthia; Vageli, Dimitra; Kaisaridou, Despina; Nakou, Marianna; Netsika, Maria; Vladica, Natalia; Daponte, Alexandros; Koukoulis, George

    2007-12-01

    The routine assessment of HER-2 expression can be affected by many immunohistological preanalytical and analytical variables. The evaluation of non-informative HER-2 tests, because of 2(+) scores, has been addressed in studies using in situ hybridization (fluorescent in situ hybridization (FISH) or chromogenic in situ hybridization (CISH)). There are very few studies that additionally checked 2(+) cases by quantitative reverse transcription-PCR (QRT-PCR). We analyzed totally 195 breast carcinoma cases, 70 of them showing 2(+) immunoreaction, with FISH/CISH and QRT-PCR. Confirmed amplification in 2(+) cases fell within the reported range (12.8% vs. 8-44%) and some of them showed lower mRNA levels indicating a genuine decrease of HER-2 protein as a mechanism for the non-informative score. In other cases, increased mRNA levels could be ascribed to HER-2 polysomy, verifying previous observations of immunohistologically detectable HER-2 polysomy. A remarkable subset of the 2(+) cases showed "normal" mRNA levels without amplification or polysomy and technical parameters as well as heterogeneity could be incriminated. The overall concordance of QRT-PCR and FISH was 93.8%, highest than most previously reported. Yet, the lack of clear cut-off mRNA values and the challenge of sample microdissection hinder QRT-PCR from claiming the status of a gold standard test for HER-2 evaluation.

  18. Nonnative Fishes in the Upper Mississippi River System

    Science.gov (United States)

    Irons, Kevin S.; DeLain, Steven A.; Gittinger, Eric; Ickes, Brian S.; Kolar, Cindy S.; Ostendort, David; Ratcliff, Eric N.; Benson, Amy J.; Irons, Kevin S.

    2009-01-01

    The introduction, spread, and establishment of nonnative species is widely regarded as a leading threat to aquatic biodiversity and consequently is ranked among the most serious environmental problems facing the United States today. This report presents information on nonnative fish species observed by the Long Term Resource Monitoring Program on the Upper Mississippi River System a nexus of North American freshwater fish diversity for the Nation. The Long Term Resource Monitoring Program, as part of the U.S. Army Corps of Engineers' Environmental Management Plan, is the Nation's largest river monitoring program and stands as the primary source of standardized ecological information on the Upper Mississippi River System. The Long Term Resource Monitoring Program has been monitoring fish communities in six study areas on the Upper Mississippi River System since 1989. During this period, more than 3.5 million individual fish, consisting of 139 species, have been collected. Although fish monitoring activities of the Long Term Resource Monitoring Program focus principally on entire fish communities, data collected by the Program are useful for detecting and monitoring the establishment and spread of nonnative fish species within the Upper Mississippi River System Basin. Sixteen taxa of nonnative fishes, or hybrids thereof, have been observed by the Long Term Resource Monitoring Program since 1989, and several species are presently expanding their distribution and increasing in abundance. For example, in one of the six study areas monitored by the Long Term Resource Monitoring Program, the number of established nonnative species has increased from two to eight species in less than 10 years. Furthermore, contributions of those eight species can account for up to 60 percent of the total annual catch and greater than 80 percent of the observed biomass. These observations are critical because the Upper Mississippi River System stands as a nationally significant pathway for

  19. Molecular confirmation of hybridization between Dascyllus reticulatus × Dascyllus aruanus from the Great Barrier Reef

    KAUST Repository

    He, Song; Johansen, Jacob L.; Hoey, Andrew S.; Pappas, Melissa; Berumen, Michael L.

    2017-01-01

    To date, more than 81 species of tropical coral reef fish have been reported to hybridize in nature, spanning multiple families, including the Chaetodontidae, Pomacanthidae, and Labridae. Hybridization, however, is seemingly rare among benthic nesting species that engage in pair spawning, such as the Pomacentridae. Here, we present evidence for the first molecularly confirmed record of hybridization within the genus Dascyllus; D. aruanus and D. reticulatus. Interestingly, although many hybridization events are attributed to peripheral range effects or areas of limited overlap among otherwise allopatric species, this hybrid individual was collected from the northern Great Barrier Reef, centrally located within the distribution ranges of both species. The hybrid exhibited coloration and meristic counts intermediate between D. aruanus and D. reticulatus. Diagnostic genetic markers and subsequent microsatellites analysis confirmed that this individual was a hybrid offspring of D. aruanus and D. reticulatus, with the latter providing the maternal contribution. The occurrence of the D. aruanus × D. reticulatus hybrid on the Great Barrier Reef represents an exception to the otherwise species-specific haplotypes. The nuclear diagnostic marker which was identified during this study could serve as a hybrid indicator and benefit future hybrid investigations for hybridization between these two species.

  20. Molecular confirmation of hybridization between Dascyllus reticulatus × Dascyllus aruanus from the Great Barrier Reef

    KAUST Repository

    He, Song

    2017-11-17

    To date, more than 81 species of tropical coral reef fish have been reported to hybridize in nature, spanning multiple families, including the Chaetodontidae, Pomacanthidae, and Labridae. Hybridization, however, is seemingly rare among benthic nesting species that engage in pair spawning, such as the Pomacentridae. Here, we present evidence for the first molecularly confirmed record of hybridization within the genus Dascyllus; D. aruanus and D. reticulatus. Interestingly, although many hybridization events are attributed to peripheral range effects or areas of limited overlap among otherwise allopatric species, this hybrid individual was collected from the northern Great Barrier Reef, centrally located within the distribution ranges of both species. The hybrid exhibited coloration and meristic counts intermediate between D. aruanus and D. reticulatus. Diagnostic genetic markers and subsequent microsatellites analysis confirmed that this individual was a hybrid offspring of D. aruanus and D. reticulatus, with the latter providing the maternal contribution. The occurrence of the D. aruanus × D. reticulatus hybrid on the Great Barrier Reef represents an exception to the otherwise species-specific haplotypes. The nuclear diagnostic marker which was identified during this study could serve as a hybrid indicator and benefit future hybrid investigations for hybridization between these two species.

  1. Cross-species BAC-FISH painting of the tomato and potato chromosome 6 reveals undescribed chromosomal rearrangements

    NARCIS (Netherlands)

    Tang, X.; Szinay, D.; Ramanna, M.S.; Vossen, van der E.A.G.; Datema, E.; Klein Lankhorst, R.M.; Boer, de J.M.; Peters, S.A.; Bachem, C.W.B.; Stiekema, W.J.; Visser, R.G.F.; Jong, de J.H.; Bai, Y.

    2008-01-01

    Ongoing genomics projects of tomato (Solanum lycopersicum ) and potato (Solanum tuberosum) are providing unique tools for comparative mapping studies in Solanaceae. At the chromosomal level, BACs can be positioned on pachytene comple-ments by fluorescent in situ hybridization (FISH) on homoeologous

  2. Formation of diploid and triploid hybrid groupers (hybridization of Epinephelus coioides ♀ × Epinephelus lanceolatus ♂) and their 5S gene analysis.

    Science.gov (United States)

    Huang, Wen; Qin, Qinbo; Yang, Huirong; Li, Shuisheng; Hu, Chaoqun; Wang, Yude; Zhang, Yong; Liu, Shaojun; Lin, Haoran

    2016-10-07

    Interspecies hybridization is widely used to achieve heterosis or hybrid vigor, which has been observed and harnessed by breeders for centuries. Natural allopolyploid hybrids generally exhibit more superior heterosis than both the diploid progenies and their parental species. However, polyploid formation processes have been long ignored, the genetic basis of heterosis in polyploids remains elusive. In the present study, triploid hybrids had been demonstrated to contain two sets of chromosomes from mother species and one set from father species. Cellular polyploidization process in the embryos had been traced. The triploid hybrids might be formed by failure formation of the second polarized genome during the second meiosis stage. Four spindle centers were observed in anaphase stage of the first cell division. Three spindle centers were observed in side of cell plate after the first cell division. The 5S rDNA genes of four types of groupers were cloned and analyzed. The diploid and triploid hybrids had been proved to contain the tandem chimera structures which were recombined by maternal and paternal monomer units. The results indicated that genome re-fusion had occurred in the hybrid progenies. To further elucidate the genetic patterns of diploid and triploid hybrids, fluorescence chromosome location had been carried out, maternal 5S gene (M-386) were used as the probe. The triploid hybrids contained fewer fluorescence loci numbers than the maternal species. The results indicated that participation of paternal 5S gene in the triploid hybrid genome had degraded the match rates of M-386 probe. Our study is the first to investigate the cellular formation processes of natural allopolyploids in hybrid fish, the cellular polyploidization process may be caused by failure formation of the second polarized genome during the meiosis, and our results will provide the molecular basis of hybrid vigor in interspecies hybridization.

  3. Swimming Performance of Toy Robotic Fish

    Science.gov (United States)

    Petelina, Nina; Mendelson, Leah; Techet, Alexandra

    2015-11-01

    HEXBUG AquaBotsTM are a commercially available small robot fish that come in a variety of ``species''. These models have varying caudal fin shapes and randomly-varied modes of swimming including forward locomotion, diving, and turning. In this study, we assess the repeatability and performance of the HEXBUG swimming behaviors and discuss the use of these toys to develop experimental techniques and analysis methods to study live fish swimming. In order to determine whether these simple, affordable model fish can be a valid representation for live fish movement, two models, an angelfish and a shark, were studied using 2D Particle Image Velocimetry (PIV) and 3D Synthetic Aperture PIV. In a series of experiments, the robotic fish were either allowed to swim freely or towed in one direction at a constant speed. The resultant measurements of the caudal fin wake are compared to data from previous studies of a real fish and simplified flapping propulsors.

  4. GROWTH PERFORMANCE AND FEED CONVERSION RATIO (FCR OF HYBRID FINGERLINGS (CATLA CATLA X LABEO ROHITA FED ON COTTONSEED MEAL, SUNFLOWER MEAL AND BONE MEAL

    Directory of Open Access Journals (Sweden)

    T. SAHZADI, M. SALIM, UM-E-KALSOOM AND K. SHAHZAD

    2006-10-01

    Full Text Available An experiment was conducted in six glass aquaria to study the growth performance and feed conversion ratio (FCR of hybrid fingerlings (Catla catla x Labeo rohita fed on sunflower meal, cottonseed meal and bone meal. Two replicates for each ingredient were followed. The feed was supplied at the rate of 4% of wet body weight of fingerlings twice a day. The hybrid (Catla catla x Labeo rohita fingerlings gained highest body weight (1.62 ± 0.0 g on sunflower meal, followed by cottonseed meal (1.61 ± 0.01 g and bone meal (1.52 ± 0.0 g. The total length obtained by hybrid fish was 6.35 ± 0.05 cm on sunflower meal, 6.12 ± 0.05 cm on cottonseed meal and 5.85 ± 0.05 cm on bone meal. The overall mean values of FCR were lower (better on sunflower meal (1.78 ± 0.05, followed by cottonseed meal (2.17 ± 0.01 and bone meal (2.46 ± 0.01. Thus, The sunflower meal and cottonseed meal, on the basis of growth performance and better FCR, can be included in the feed formulation for hybrid fingerlings.

  5. Hybrid Taxis Give Fuel Economy a Lift -Clean Cities Fleet Experiences -

    Energy Technology Data Exchange (ETDEWEB)

    None

    2009-04-01

    The hybrid taxis are able to achieve about twice the gas mileage of a conventional taxi while helping cut gasoline use and fuel costs. Tax credits and other incentives are helping both company owners and drivers make the switch to hybrids.

  6. Fish irradiation

    International Nuclear Information System (INIS)

    Kovacs, J.; Tengumnuay, C.; Juangbhanich, C.

    1970-01-01

    Chub-mackerel was chosen for the study because they are the most common fish in Thailand. Preliminary investigations were conducted to determine the maximum radiation dose of gamma-rays by organoleptic tests. The samples were subjected to radiation at various doses up to 4 Mrad. Many experiments were conducted using other kinds of fish. The results showed that 1 Mrad would be the maximum acceptable dose for fish. Later, the influence of the radiation dose from 0.1-1 Mrad was studied in order to find the optimum acceptable dose for preservation of fish without off-flavour. For this purpose, the Hedonic scale was used. It was found that 0.2 and 0.5 Mrad gave the best result on Chub mackerel. The determinations of optimum dose, organoleptic, microbiological and trimethylamine content changes were done. The results showed that Chub mackerel irradiated at 0.2, 0.5 and 1 Mrad stored at 3 0 C for 71 days were still acceptable, on the contrary the untreated samples were found unacceptable at 14 days. The trimethylamine increment was significantly higher in the untreated samples. At 15 days storage, trimethylamine in the non-irradiated Chub-mackerel was about 10 times higher than the irradiated ones. At 51 and 79 days storage, about 13 times higher in the control samples than the irradiated samples except 0.1 Mrad. Only 2 times higher was found for the 0.1 Mrad. The microbiological results showed that the irradiation above 0.2 Mrad gave favorable extension of shelf-life of fish

  7. Reversed sex change by widowed males in polygynous and protogynous fishes: female removal experiments in the field

    Science.gov (United States)

    Kuwamura, Tetsuo; Suzuki, Shohei; Kadota, Tatsuru

    2011-12-01

    Sex change, either protogyny (female to male) or protandry (male to female), is well known among fishes, but evidence of bidirectional sex change or reversed sex change in natural populations is still very limited. This is the first report on female removal experiments for polygnous and protogynous fish species to induce reversed sex change in the widowed males in the field. We removed all of the females and juveniles from the territories of dominant males in the cleaner wrasse Labroides dimidiatus (Labridae) and the rusty angelfish Centropyge ferrugata (Pomacanthidae) on the coral reefs of Okinawa. In both species, if new females or juveniles did not immigrate into the territories of the widowed males, some of them emigrated to form male-male pairs. When a male-male pair formed, the smaller, subordinate partner began to perform female sexual behaviours ( n = 4 in L. dimidiatus; n = 2 in C. ferrugata) and, finally, released eggs ( n = 1, respectively). Thus, the reversed sex change occurred in the widowed males according to the change of their social status. These results suggest that such female removal experiments will contribute to the discovery of reversed sex change in the field also in other polygnous and protogynous species.

  8. 50 CFR 23.43 - What are the requirements for a wildlife hybrid?

    Science.gov (United States)

    2010-10-01

    ... 50 Wildlife and Fisheries 6 2010-10-01 2010-10-01 false What are the requirements for a wildlife hybrid? 23.43 Section 23.43 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF... IMPORTATION OF WILDLIFE AND PLANTS (CONTINUED) CONVENTION ON INTERNATIONAL TRADE IN ENDANGERED SPECIES OF WILD...

  9. Spatial segregation of spawning habitat limits hybridization between sympatric native Steelhead and Coastal Cutthroat Trout

    Science.gov (United States)

    Buehrens, T.W.; Glasgow, J.; Ostberg, Carl O.; Quinn, T.P.

    2013-01-01

    Native Coastal Cutthroat Trout Oncorhynchus clarkii clarkii and Coastal Steelhead O. mykiss irideus hybridize naturally in watersheds of the Pacific Northwest yet maintain species integrity. Partial reproductive isolation due to differences in spawning habitat may limit hybridization between these species, but this process is poorly understood. We used a riverscape approach to determine the spatial distribution of spawning habitats used by native Coastal Cutthroat Trout and Steelhead as evidenced by the distribution of recently emerged fry. Molecular genetic markers were used to classify individuals as pure species or hybrids, and individuals were assigned to age-classes based on length. Fish and physical habitat data were collected in a spatially continuous framework to assess the relationship between habitat and watershed features and the spatial distribution of parental species and hybrids. Sampling occurred in 35 reaches from tidewaters to headwaters in a small (20 km2) coastal watershed in Washington State. Cutthroat, Steelhead, and hybrid trout accounted for 35%, 42%, and 23% of the fish collected, respectively. Strong segregation of spawning areas between Coastal Cutthroat Trout and Steelhead was evidenced by the distribution of age-0 trout. Cutthroat Trout were located farther upstream and in smaller tributaries than Steelhead were. The best predictor of species occurrence at a site was the drainage area of the watershed that contributed to the site. This area was positively correlated with the occurrence of age-0 Steelhead and negatively with the presence of Cutthroat Trout, whereas hybrids were found in areas occupied by both parental species. A similar pattern was observed in older juveniles of both species but overlap was greater, suggesting substantial dispersal of trout after emergence. Our results offer support for spatial reproductive segregation as a factor limiting hybridization between Steelhead and Coastal Cutthroat Trout.

  10. Detection of iron-depositing Pedomicrobium species in native biofilms from the Odertal National Park by a new, specific FISH probe.

    Science.gov (United States)

    Braun, Burga; Richert, Inga; Szewzyk, Ulrich

    2009-10-01

    Iron-depositing bacteria play an important role in technical water systems (water wells, distribution systems) due to their intense deposition of iron oxides and resulting clogging effects. Pedomicrobium is known as iron- and manganese-oxidizing and accumulating bacterium. The ability to detect and quantify members of this species in biofilm communities is therefore desirable. In this study the fluorescence in situ hybridization (FISH) method was used to detect Pedomicrobium in iron and manganese incrusted biofilms. Based on comparative sequence analysis, we designed and evaluated a specific oligonucleotide probe (Pedo 1250) complementary to the hypervariable region 8 of the 16S rRNA gene for Pedomicrobium. Probe specificities were tested against 3 different strains of Pedomicrobium and Sphingobium yanoikuyae as non-target organism. Using optimized conditions the probe hybridized with all tested strains of Pedomicrobium with an efficiency of 80%. The non-target organism showed no hybridization signals. The new FISH probe was applied successfully for the in situ detection of Pedomicrobium in different native, iron-depositing biofilms. The hybridization results of native bioflims using probe Pedo_1250 agreed with the results of the morphological structure of Pedomicrobium bioflims based on scanning electron microscopy.

  11. The influence of fish feed pellets on the stability of seabed sediment: A laboratory flume investigation

    Science.gov (United States)

    Neumeier, Urs; Friend, Patrick L.; Gangelhof, Uffe; Lunding, Jens; Lundkvist, Morten; Bergamasco, Alessandro; Amos, Carl L.; Flindt, Mogens

    2007-11-01

    Superfluous fish food settling below fish farms can have a negative impact on the seabed. To aid in the assessment of this impact a series of flume experiments, designed to mimic seabed conditions below a fish farm, was conducted with the aim of examining the effects of fish pellets on the stability of fine sediments. Artificial beds, with varying quantities of fish pellets incorporated both within the sediment matrix and lying on the sediment surface, were allowed to consolidate for different periods of time ranging from 1 to 10 days, and then subjected to erosion experiments. In flume experiments containing fish pellets, a bacterial biofilm developed at the sediment-water interface after a few days. In the control experiments (no fish pellets), a diatom biofilm caused extensive stabilisation of the surface sediment. The erosion experiments showed that the addition of fish pellets reduced the surface erosion threshold by more than 50%. The stability decrease was more pronounced in the experiments with greater amounts of pellets. Evidence of drag reduction due to high suspended sediment concentration was also observed. This phenomenon is discussed and a correction formula is proposed for the effective shear stress experienced by the bed.

  12. [Cytogenetic Abnormalities and Outcomes of 117 Patients with Multiple Myeloma Detected by FISH].

    Science.gov (United States)

    Zhai, Bing; Zou, Dan-Dan; Yan, Jian-Jun; Wang, Nan; Wang, Li-Li; Zhu, Hong-Li; Huang, Wen-Rong; Yu, Li

    2016-02-01

    To analyze the cytogenetic abnormalities and prognostic outcomes of patients with multiple myeloma (MM) detected by fluorescence in situ hybridization (FISH). The clinical record of 117 newly-diagnosed patients with MM treated in department of hematology and geriatric hematology of our hospital for 7 years were collected, and their molecular cytogenetic abnormalities detected by FISH and the clinical outcome were analyzed retrospectively. The detected rate of cytogenetic abnormality was 76.9%(90/117), the most common abnormality deteted by FISH was 1q21+ (71.1%), followed by 13q- (56.6%). The cross comparison method showed that 13q- and 17p13-, t(11;14) and t(4;14) were related respectively. All the patients with cytogenetic abnormalities showed no significant difference in the overall survival from cytogenetic normal patients. The positive rate of molecular cytogenetic abnormalities detected by FISH in MM patients is high, but data from larger and longer studies are needed to evaluate the prognostic outcomes.

  13. [Chromosomal localization of foreign genes in transgenic mice using dual-color fluorescence in situ hybridization].

    Science.gov (United States)

    Lin, Dan; Gong, Xiu-li; Li, Wei; Guo, Xin-bing; Zhu, Yi-wen; Huang, Ying

    2008-02-01

    To establish a highly sensitive and specific dual-color fluorescence in situ hybridization (D-FISH) method used for chromosomal localization of foreign genes in double transgenic mice. Two strains of double transgenic mice were used in this experiment, one was integrated with the herpes simplex virus thymidine kinase (HSV-tk) and the enhanced green fluorescence protein (eGFP), the other was with the short hairpin RNA interference(RNAi) and beta(654). Splenic cells cultured in vitro were arrested in metaphase by colchicine and hybridized with digoxigenin-labeled and biotinylated DNA probes, then detected by rhodamine-conjugated avidin and FITC-conjugated anti-digoxigenin. Dual-color fluorescence signals were detected on the same metaphase in both transgenic mice strains. In HSV-tk/eGFP double transgenic mice, strong green fluorescence for HSV-tk and red for eGFP were observed and localized at 2E5-G3 and 8A2-A4 respectively. In beta(654)/RNAi mice, beta(654) was detected as red fluorescence on chromosome 7D3-E2, and RNAi showed random integration on chromosomes. It was detected as green fluorescence on chromosome 12B1 in one mouse, while on 1E2.3-1F and 3A3 in the other. Highly sensitive and specific D-FISH method was established using the self-prepared DNA probes, and chromosomal localization of the foreign genes was also performed in combination with G-banding in double transgenic mice. This technology will facilitate the researches in transgenic animals and gene therapy models.

  14. Rapid identification of bacteria and candida using pna-fish from blood and peritoneal fluid cultures: a retrospective clinical study

    Directory of Open Access Journals (Sweden)

    Harris Dana M

    2013-01-01

    Full Text Available Abstract Background Peptide nucleic acid fluorescent in situ hybridization (PNA-FISH is a rapid and established method for identification of Candida sp., Gram positive, and Gram negative bacteria from positive blood cultures. This study reports clinical experience in the evaluation of 103 positive blood cultures and 17 positive peritoneal fluid cultures from 120 patients using PNA-FISH. Our study provides evidence as to potential pharmaceutical cost savings based on rapid pathogen identification, in addition to the novel application of PNA-FISH to peritoneal fluid specimens. Methods Identification accuracy and elapsed time to identification of Gram positives, Gram negatives, and Candida sp., isolated from blood and peritoneal fluid cultures were assessed using PNA-FISH (AdvanDx, as compared to standard culture methods. Patient charts were reviewed to extrapolate potential pharmaceutical cost savings due to adjustment of antimicrobial or antifungal therapy, based on identification by PNA-FISH. Results In blood cultures, time to identification by standard culture methods for bacteria and Candida sp., averaged 83.6 hours (95% CI 56.7 to 110.5. Identification by PNA-FISH averaged 11.2 hours (95% CI 4.8 to 17.6. Overall PNA-FISH identification accuracy was 98.8% (83/84, 95% CI 93.5% to 99.9% as compared to culture. In peritoneal fluid, identification of bacteria by culture averaged 87.4 hours (95% CI −92.4 to 267.1. Identification by PNA-FISH averaged 16.4 hours (95% CI −57.3 to 90.0. Overall PNA-FISH identification accuracy was 100% (13/13, 95% CI 75.3% to 100%. For Candida sp., pharmaceutical cost savings based on PNA-FISH identification could be $377.74/day. For coagulase-negative staphylococcus (CoNS, discontinuation of vancomycin could result in savings of $20.00/day. Conclusions In this retrospective study, excellent accuracy of PNA-FISH in blood and peritoneal fluids with reduced time to identification was observed, as compared to

  15. Fishers, Fishing, and Overfishing: American Experiences in Global Perspective, 1976-2006

    OpenAIRE

    Mansel Blackford

    2009-01-01

    Since the mid-1970s, overfishing has had a considerable impact on the American seafood industry, and companies serving the American seafood market have taken steps to respond to the crisis. Following a brief survey of modern-day fishing and of the steps that have been taken to mitigate overfishing, the strategies adopted for commercial fishing in American waters, especially those in Alaska, are examined. Through their attempts to deal with the challenges posed by overfishing, fishers, seafood...

  16. Resistance of sunflower hybrids to imazamox and tribenuron-methyl

    DEFF Research Database (Denmark)

    Bozic, D; Saric, M; Malidza, G

    2012-01-01

    The response of the imazamox resistant and susceptible sunflower hybrids Rimi and S to imazamox and of tribenuron-methyl resistant and susceptible hybrids Rsu and S to tribenuron-methyl was investigated both in a whole-plant bioassay and in field experiments. Plants were treated post-emergence wi......The response of the imazamox resistant and susceptible sunflower hybrids Rimi and S to imazamox and of tribenuron-methyl resistant and susceptible hybrids Rsu and S to tribenuron-methyl was investigated both in a whole-plant bioassay and in field experiments. Plants were treated post...

  17. Integrated laboratory scale demonstration experiment of the hybrid sulphur cycle and preliminary scale-up

    International Nuclear Information System (INIS)

    Leybros, J.; Rivalier, P.; Saturnin, A.; Charton, S.

    2010-01-01

    The hybrid sulphur cycle is today one of the most promising processes to produce hydrogen on a massive scale within the scope of high temperature nuclear reactors development. Thus, the Fuel Cycle Technology Department at CEA Marcoule is involved in studying the hybrid sulphur process from a technical and economical performance standpoint. Based on mass and energy balance calculations, a ProsimPlus TM flow sheet and a commercial plant design were prepared. This work includes a study on sizing of the main equipment. The capital cost has been estimated using the major characteristics of main equipment based upon formulae and charts published in literature. A specific approach has been developed for electrolysers. Operational costs are also proposed for a plant producing 1000 mol/s H 2 . Bench scale and pilot experiments must focus on the electrochemical step due to limited experimental data. Thus, a pilot plant with a hydrogen capacity of 100 NL/h was built with the aim of acquiring technical and technological data for electrolysis. This pilot plant was designed to cover a wide range of operating conditions: sulphuric acid concentrations up to 60 wt.%, temperatures up to 100 deg. C and pressures up to 10 bar. New materials and structures recently developed for fuel cells, which are expected to yield significant performance improvements when applied to classical electrochemical processes, will be tested. All experiments will be coupled with phenomenological simulation tools developed jointly with the experimental programme. (authors)

  18. Natural selection interacts with recombination to shape the evolution of hybrid genomes.

    Science.gov (United States)

    Schumer, Molly; Xu, Chenling; Powell, Daniel L; Durvasula, Arun; Skov, Laurits; Holland, Chris; Blazier, John C; Sankararaman, Sriram; Andolfatto, Peter; Rosenthal, Gil G; Przeworski, Molly

    2018-05-11

    To investigate the consequences of hybridization between species, we studied three replicate hybrid populations that formed naturally between two swordtail fish species, estimating their fine-scale genetic map and inferring ancestry along the genomes of 690 individuals. In all three populations, ancestry from the "minor" parental species is more common in regions of high recombination and where there is linkage to fewer putative targets of selection. The same patterns are apparent in a reanalysis of human and archaic admixture. These results support models in which ancestry from the minor parental species is more likely to persist when rapidly uncoupled from alleles that are deleterious in hybrids. Our analyses further indicate that selection on swordtail hybrids stems predominantly from deleterious combinations of epistatically interacting alleles. Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

  19. Who cares about fish welfare?

    DEFF Research Database (Denmark)

    Ellingsen, Kristian; Grimsrud, Kristine; Nielsen, Hanne Marie

    2015-01-01

    . Research limitations/implications – In this study willingness to pay is measured using a hypothetical choice experiment. Values people express as citizens, however, may not accurately predict true consumer behaviour. This is generally referred to as “citizen-consumer duality” and may have affected...... about the appropriate way to pay for better welfare standards in fish production. Design/methodology/approach – On the basis of two focus group sessions, a survey questionnaire was developed and distributed to a representative sample of 2,147 Norwegian households via e-mail. Findings – Results showed...... the results. Practical implications – The study shows that there is a national market for welfare-assured fish products, but education initiatives focusing on fish farming and fish welfare issues would further influence the attitudes and purchasing habits of Norwegian consumers. Originality/value – Although...

  20. A biologically inspired artificial fish using flexible matrix composite actuators: analysis and experiment

    International Nuclear Information System (INIS)

    Zhang, Zhiye; Philen, Michael; Neu, Wayne

    2010-01-01

    A bio-inspired prototype fish using the flexible matrix composite (FMC) muscle technology for fin and body actuation is developed. FMC actuators are pressure driven muscle-like actuators capable of large displacements as well as large blocking forces. An analytical model of the artificial fish using FMC actuators is developed and analysis results are presented. An experimental prototype of the artificial fish having FMC artificial muscles has been completed and tested. Constant mean thrusts have been achieved in the laboratory for a stationary fish for different undulation frequencies around 1 Hz. The experimental results demonstrate that a nearly constant thrust can be achieved through tuning of excitation frequency for given body stiffness. Free swimming results show that the prototype can swim at approximately 0.3 m s −1

  1. The virtual lover: variable and easily guided 3D fish animations as an innovative tool in mate-choice experiments with sailfin mollies-I. Design and implementation.

    Science.gov (United States)

    Müller, Klaus; Smielik, Ievgen; Hütwohl, Jan-Marco; Gierszewski, Stefanie; Witte, Klaudia; Kuhnert, Klaus-Dieter

    2017-02-01

    Animal behavior researchers often face problems regarding standardization and reproducibility of their experiments. This has led to the partial substitution of live animals with artificial virtual stimuli. In addition to standardization and reproducibility, virtual stimuli open new options for researchers since they are easily changeable in morphology and appearance, and their behavior can be defined. In this article, a novel toolchain to conduct behavior experiments with fish is presented by a case study in sailfin mollies Poecilia latipinna . As the toolchain holds many different and novel features, it offers new possibilities for studies in behavioral animal research and promotes the standardization of experiments. The presented method includes options to design, animate, and present virtual stimuli to live fish. The designing tool offers an easy and user-friendly way to define size, coloration, and morphology of stimuli and moreover it is able to configure virtual stimuli randomly without any user influence. Furthermore, the toolchain brings a novel method to animate stimuli in a semiautomatic way with the help of a game controller. These created swimming paths can be applied to different stimuli in real time. A presentation tool combines models and swimming paths regarding formerly defined playlists, and presents the stimuli onto 2 screens. Experiments with live sailfin mollies validated the usage of the created virtual 3D fish models in mate-choice experiments.

  2. The development of FISH tools for genetic, phylogenetic and breeding studies in tomato (Solanum lycopersicum)

    NARCIS (Netherlands)

    Szinay, D.

    2010-01-01

    In this thesis various fluorescence in situ hybridization (FISH) technologies are described to support genome projects, plant breeding and phylogenetic analysis on tomato (Solanum lycopersicum, 2n=24). Its genome is 980 Mb and only 30 % are single copy sequences, which are mostly found in the

  3. Influence of habitat degradation on fish replenishment

    Science.gov (United States)

    McCormick, M. I.; Moore, J. A. Y.; Munday, P. L.

    2010-09-01

    Temperature-induced coral bleaching is a major threat to the biodiversity of coral reef ecosystems. While reductions in species diversity and abundance of fish communities have been documented following coral bleaching, the mechanisms that underlie these changes are poorly understood. The present study examined the impacts of coral bleaching on the early life-history processes of coral reef fishes. Daily monitoring of fish settlement patterns found that ten times as many fish settled to healthy coral than sub-lethally bleached coral. Species diversity of settling fishes was least on bleached coral and greatest on dead coral, with healthy coral having intermediate levels of diversity. Laboratory experiments using light-trap caught juveniles showed that different damselfish species chose among healthy, bleached and dead coral habitats using different combinations of visual and olfactory cues. The live coral specialist, Pomacentrus moluccensis, preferred live coral and avoided bleached and dead coral, using mostly visual cues to inform their habitat choice. The habitat generalist, Pomacentrus amboinensis, also preferred live coral and avoided bleached and dead coral but selected these habitats using both visual and olfactory cues. Trials with another habitat generalist, Dischistodus sp., suggested that vision played a significant role. A 20 days field experiment that manipulated densities of P. moluccensis on healthy and bleached coral heads found an influence of fish density on juvenile weight and growth, but no significant influence of habitat quality. These results suggests that coral bleaching will affect settlement patterns and species distributions by influencing the visual and olfactory cues that reef fish larvae use to make settlement choices. Furthermore, increased fish density within the remaining healthy coral habitats could play an important role in influencing population dynamics.

  4. Little Fish in a Big Pond--Time to Get Schooled!

    Science.gov (United States)

    Moye, Johnny J.

    2011-01-01

    One technique used by many who fish is to catch a fish in a creek or pond and then release it in a different pond. This satisfies the desire for sport, and it also serves to stock a pond that may need replenishment. Of course this restocking can be a traumatic experience for the new fish. To survive in the new environment, the fish must find its…

  5. Determination of HER2 gene amplification by chromogenic in situ hybridization (CISH) in archival breast carcinoma.

    Science.gov (United States)

    Zhao, Jianxin; Wu, Rina; Au, Alfred; Marquez, Abbey; Yu, Yibing; Shi, Zuorong

    2002-06-01

    To compare the efficacy of chromogenic in situ hybridization (CISH(TM)) with fluorescence in situ (FISH) hybridization and immunohistochemistry (IHC) in determination of the HER2 status in human breast cancer. HER2 gene amplification was determined on formalin-fixed paraffin-embedded (FFPE) sections of 62 invasive breast cancers by FISH and followed by CISH using a digoxigenin (DIG)-labeled HER2 DNA probe generated by Subtraction Probe Technology (SPT(TM)), and a biotin-labeled chromosome 17 centromeric (chr.17cen) probe. The sections were heat treated and enzyme digested. After in situ hybridization, the HER2 probe was detected with fluorescein (FITC)-anti-DIG for FISH, followed by peroxidase-anti-FITC and diaminobenzidine (DAB) for CISH. The chr.17cen probe was detected with peroxidase-streptavidin and DAB. For CISH application, HER2 gene copies or chromosome 17 centromeres and morphology of cells were easily visualized simultaneously with a 40x objective under bright-field microscope in hematoxylin-counterstained sections. IHC study of HER2 overexpression was performed on adjacent sections using a panel of three HER2 antibodies (TAB 250, CB11, A0485), and staining was scored according to the criteria specified in the HercepTest. HER2 gene amplification detected by CISH was visualized typically as large DAB-stained clusters or by many dots in the nucleus. FISH and CISH identified HER2 gene amplification in 19% of the tumors. Chromosome 17 polysomy was detected in 31% of the tumors. HER2 overexpression was demonstrated in 19% (TAB 250), 23% (CB11), and 36% (A0485) of the tumors. Complete concordance between the results of CISH with FISH, TAB 250, CB11, and A0485 was seen in 100%, 97%, 94%, and 84% of the cases, respectively. By permitting observation of morphology using a bright-field microscope, CISH is an accurate, practical, and economical approach to screen HER2 status in breast cancers. It is a useful methodology for confirming ambiguous IHC results.

  6. Multifractal-based nuclei segmentation in fish images.

    Science.gov (United States)

    Reljin, Nikola; Slavkovic-Ilic, Marijeta; Tapia, Coya; Cihoric, Nikola; Stankovic, Srdjan

    2017-09-01

    The method for nuclei segmentation in fluorescence in-situ hybridization (FISH) images, based on the inverse multifractal analysis (IMFA) is proposed. From the blue channel of the FISH image in RGB format, the matrix of Holder exponents, with one-by-one correspondence with the image pixels, is determined first. The following semi-automatic procedure is proposed: initial nuclei segmentation is performed automatically from the matrix of Holder exponents by applying predefined hard thresholding; then the user evaluates the result and is able to refine the segmentation by changing the threshold, if necessary. After successful nuclei segmentation, the HER2 (human epidermal growth factor receptor 2) scoring can be determined in usual way: by counting red and green dots within segmented nuclei, and finding their ratio. The IMFA segmentation method is tested over 100 clinical cases, evaluated by skilled pathologist. Testing results show that the new method has advantages compared to already reported methods.

  7. possible stimuli for social behaviour in three species of freshwater fish

    African Journals Online (AJOL)

    Social behaviour in fish is manifested in many situations and one form. of this phenomenon is especially interesting. Some species of fish have .... Experi- mental methods of study of fish behaviour, in Techniques lor the Investigation 0/ Fish. Physiology, ed. Pavlovskii, E. N., Oldbourne. Press, London. R ep rod u ced b y Sa b.

  8. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    Directory of Open Access Journals (Sweden)

    F.E. Rosa

    Full Text Available Human epidermal growth factor receptor 2 (HER2 has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC. HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH in moderate immunoexpression (IHC 2+ cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH, which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

  9. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    Directory of Open Access Journals (Sweden)

    F.E. Rosa

    2013-03-01

    Full Text Available Human epidermal growth factor receptor 2 (HER2 has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC. HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH in moderate immunoexpression (IHC 2+ cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH, which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

  10. Specific Fluorescence in Situ Hybridization (FISH) Test to Highlight Colonization of Xylem Vessels by Xylella fastidiosa in Naturally Infected Olive Trees (Olea europaea L.)

    Science.gov (United States)

    Cardinale, Massimiliano; Luvisi, Andrea; Meyer, Joana B.; Sabella, Erika; De Bellis, Luigi; Cruz, Albert C.; Ampatzidis, Yiannis; Cherubini, Paolo

    2018-01-01

    The colonization behavior of the Xylella fastidiosa strain CoDiRO, the causal agent of olive quick decline syndrome (OQDS), within the xylem of Olea europaea L. is still quite controversial. As previous literature suggests, even if xylem vessel occlusions in naturally infected olive plants were observed, cell aggregation in the formation of occlusions had a minimal role. This observation left some open questions about the whole behavior of the CoDiRO strain and its actual role in OQDS pathogenesis. In order to evaluate the extent of bacterial infection in olive trees and the role of bacterial aggregates in vessel occlusions, we tested a specific fluorescence in situ hybridization (FISH) probe (KO 210) for X. fastidiosa and quantified the level of infection and vessel occlusion in both petioles and branches of naturally infected and non-infected olive trees. All symptomatic petioles showed colonization by X. fastidiosa, especially in the larger innermost vessels. In several cases, the vessels appeared completely occluded by a biofilm containing bacterial cells and extracellular matrix and the frequent colonization of adjacent vessels suggested a horizontal movement of the bacteria. Infected symptomatic trees had 21.6 ± 10.7% of petiole vessels colonized by the pathogen, indicating an irregular distribution in olive tree xylem. Thus, our observations point out the primary role of the pathogen in olive vessel occlusions. Furthermore, our findings indicate that the KO 210 FISH probe is suitable for the specific detection of X. fastidiosa. PMID:29681910

  11. Cloning and characterization of the gene for L-amino acid oxidase in hybrid tilapia.

    Science.gov (United States)

    Shen, Yubang; Fu, Gui Hong; Liu, Feng; Yue, Gen Hua

    2015-12-01

    Tilapia is the common name for a group of cichlid fishes. Identification of DNA markers significantly associated with important traits in candidate genes may speed up genetic improvement. L-Amino acid oxidase (LAO) plays a crucial role in the innate immune defences of animals. Previously, whether LAO variants were associated with economic traits had not been studied in fish. We characterized the cDNA sequence of the LAO gene of hybrid tilapia (Oreochromis spp.). Its ORF was 1536 bp, encoding a flavoenzyme of 511 amino acids. This gene consisted of seven exons and six introns. Its expression was detected in the intestine, blood, kidney, skin, liver. It was highly expressed in the intestine. After a challenge with a bacterial pathogen, Streptococcus agalactiae, its expression was up-regulated significantly in the liver, intestine and spleen (P tilapia. The investigation of relationship between polymorphism of LAO gene and disease resistance and growth in tilapia showed that one SNP was associated significantly with body length. Further experiments on whether SNPs in the LAO gene are associated with growth in tilapia and other populations could be useful in understanding more functions of the LAO gene.

  12. Fish condensate as effective replacer of fish meal protein in diet for striped snakehead, Channa striata (Bloch).

    Science.gov (United States)

    Wattanakul, Wattana; Wattanakul, Uraiwan; Thongprajukaew, Karun; Muenpo, Chutchawan

    2017-02-01

    The optimal protein replacement of fish meal (FM) by fish condensate (FC) was investigated in striped snakehead, Channa striata (Bloch) (1.78 ± 0.02 g initial weight). The FM-based diet (0FC) was replaced by substituting protein from FC for 100 (100FC), 200 (200FC), 300 (300FC), 400 (400FC), 500 (500FC) or 600 (600FC) g kg -1 of the FM, and a commercial diet (CD) for carnivorous fish was included for comparison. The experiment was conducted indoors under completely randomized design (8 treatments × 3 replications × 60 fish per pond) over a 6-month trial. There were no significant differences in water quality during the experiment. The fish fed with 500FC had superior growth performance and feed utilization. This dietary treatment gave similar levels to all observed specific activities of digestive enzymes as did baseline 0FC. Survival, carcass composition, hematological parameters and liver histopathology were not negatively impacted by this protein replacement level. Economic analysis also supports the use of this by-product as a potent protein replacer in striped snakehead diet. Findings from the current study indicate that a 500 g kg -1 protein replacement of FM by FC is near optimal for striped snakehead, and similar use of it in the aquafeed of other species appears worth further studies.

  13. Stable Composition of the Nano- and Picoplankton Community during the Ocean Iron Fertilization Experiment LOHAFEX

    KAUST Repository

    Thiele, Stefan

    2014-11-17

    The iron fertilization experiment LOHAFEX was conducted in a cold-core eddy in the Southern Atlantic Ocean during austral summer. Within a few days after fertilization, a phytoplankton bloom developed dominated by nano- and picoplankton groups. Unlike previously reported for other iron fertilization experiments, a diatom bloom was prevented by iron and silicate co-limitation. We used 18S rRNA gene tag pyrosequencing to investigate the diversity of these morphologically similar cell types within the nano- and picoplankton and microscopically enumerated dominant clades after catalyzed reported deposition fluorescence in situ hybridization (CARD-FISH) with specific oligonucleotide probes. In addition to Phaeocystis, members of Syndiniales group II, clade 10–11, and the Micromonas clades ABC and E made up a major fraction of the tag sequences of the nano- and picoplankton community within the fertilized patch. However, the same clades were also dominant before the bloom and outside the fertilized patch. Furthermore, only little changes in diversity could be observed over the course of the experiment. These results were corroborated by CARD-FISH analysis which confirmed the presence of a stable nano- and picoplankton community dominated by Phaeocystis and Micromonas during the entire course of the experiment. Interestingly, although Syndiniales dominated the tag sequences, they could hardly be detected by CARD-FISH, possibly due to the intracellular parasitic life style of this clade. The remarkable stability of the nano- and picoplankton community points to a tight coupling of the different trophic levels within the microbial food web during LOHAFEX.

  14. Deep electroproduction of exotic hybrid mesons

    International Nuclear Information System (INIS)

    Anikin, I.V.; Pire, B.; Szymanowski, L.; Teryaev, O.V.; Wallon, S.

    2004-01-01

    We evaluate the leading order amplitude for the deep exclusive electroproduction of an exotic hybrid meson in the Bjorken regime. We show that, contrarily to naive expectation, this amplitude factorizes at the twist 2 level and thus scales like usual meson electroproduction when the virtual photon and the hybrid meson are longitudinally polarized. Exotic hybrid mesons may thus be studied in electroproduction experiments at JLAB, HERA (HERMES) or CERN (Compass)

  15. Radiobiological studies with marine fish

    International Nuclear Information System (INIS)

    Pentreath, R.J.

    1975-01-01

    The experimental methodology employed in radiobiological studies with fish is discussed and reviewed. The problems of care and maintenance of healthy stock fish are cons. (author)idered, including the techniques of egg and larval rearing. A variety of methods have been used to study the accumulation and loss of radionuclides, including labelled water, food and injections, and their relative merits are discussed in conjunction with the parameters affecting these processes. Other, more specialized, techniques that aid the physiological interpretation of tracer experiments are also discussed. Finally, consideration is given to some of the mathematical models that have been applied to radiobiological studies with fish, and of their value in extrapolating laboratory data to environmental conditions

  16. ZyFISH: a simple, rapid and reliable zygosity assay for transgenic mice.

    Directory of Open Access Journals (Sweden)

    Donal McHugh

    Full Text Available Microinjection of DNA constructs into fertilized mouse oocytes typically results in random transgene integration at a single genomic locus. The resulting transgenic founders can be used to establish hemizygous transgenic mouse lines. However, practical and experimental reasons often require that such lines be bred to homozygosity. Transgene zygosity can be determined by progeny testing assays which are expensive and time-consuming, by quantitative Southern blotting which is labor-intensive, or by quantitative PCR (qPCR which requires transgene-specific design. Here, we describe a zygosity assessment procedure based on fluorescent in situ hybridization (zyFISH. The zyFISH protocol entails the detection of transgenic loci by FISH and the concomitant assignment of homozygosity using a concise and unbiased scoring system. The method requires small volumes of blood, is scalable to at least 40 determinations per assay, and produces results entirely consistent with the progeny testing assay. This combination of reliability, simplicity and cost-effectiveness makes zyFISH a method of choice for transgenic mouse zygosity determinations.

  17. Concordance Between FISH Analysis of Her-2/Neu Gene in Breast Duct Carcinoma and Corresponding Axillary Nodal Metastases: Egyptian National Cancer Institute Experience.

    Science.gov (United States)

    Badawy, Omnia M; Hassan, Hannan; ELBakey, Heba A; Mosaad, Maha

    2018-05-10

    Breast cancer is a major health problem in Egypt. Her-2/Neu gene is routinely assessed for all breast cancer patients primarily by immunohistochemistry. At National Cancer Institute (NCI), Cairo University, Flourescence In Situ hybridization (FISH) analysis of Her-2/Neu gene is carried out for Her-2/Neu score 2 and for some cases of score 3 (particularly those assessed outside NCI). The test is performed essentially on the primary tumor. However, some situations require testing on corresponding lymph node metastases. There is a debate about the concordance between Her-2/Neu status in the primary tumor and synchronous lymph node metastases in various studies. The aim of this study was to test for the concordance between Her-2/Neu status in the primary breast tumor and corresponding axillary nodal metastases. This is a retrospective study in which FISH analysis of Her-2/Neu was carried out simultaneously on archived material of 50 cases previously diagnosed as invasive duct carcinoma and the corresponding nodal metastases from the Pathology Department, NCI. There was complete concordance between Her-2 status in the primary tumor and the corresponding axillary lymph node metastatic deposits in which Her-2 was amplified in 44% of the studied cohort of Egyptian patients. Her-2/Neu gene assessed by FISH analysis on synchronous lymph node metastases is strongly correlated with the primary tumor. Hence, it is justified to carry out the Her-2/Neu test on synchronous lymph nodes to decide on whether to carry out anti-Her-2/Neu target therapy. Further studies on other metastatic sites is recommended.

  18. Development of Thinopyrum ponticum-specific molecular markers and FISH probes based on SLAF-seq technology.

    Science.gov (United States)

    Liu, Liqin; Luo, Qiaoling; Teng, Wan; Li, Bin; Li, Hongwei; Li, Yiwen; Li, Zhensheng; Zheng, Qi

    2018-05-01

    Based on SLAF-seq, 67 Thinopyrum ponticum-specific markers and eight Th. ponticum-specific FISH probes were developed, and these markers and probes could be used for detection of alien chromatin in a wheat background. Decaploid Thinopyrum ponticum (2n = 10x = 70) is a valuable gene reservoir for wheat improvement. Identification of Th. ponticum introgression would facilitate its transfer into diverse wheat genetic backgrounds and its practical utilization in wheat improvement. Based on specific-locus-amplified fragment sequencing (SLAF-seq) technology, 67 new Th. ponticum-specific molecular markers and eight Th. ponticum-specific fluorescence in situ hybridization (FISH) probes have been developed from a tiny wheat-Th. ponticum translocation line. These newly developed molecular markers allowed the detection of Th. ponticum DNA in a variety of materials specifically and steadily at high throughput. According to the hybridization signal pattern, the eight Th. ponticum-specific probes could be divided into two groups. The first group including five dispersed repetitive sequence probes could identify Th. ponticum chromatin more sensitively and accurately than genomic in situ hybridization (GISH). Whereas the second group having three tandem repetitive sequence probes enabled the discrimination of Th. ponticum chromosomes together with another clone pAs1 in wheat-Th. ponticum partial amphiploid Xiaoyan 68.

  19. Bioinspired Smart Actuator Based on Graphene Oxide-Polymer Hybrid Hydrogels.

    Science.gov (United States)

    Wang, Tao; Huang, Jiahe; Yang, Yiqing; Zhang, Enzhong; Sun, Weixiang; Tong, Zhen

    2015-10-28

    Rapid response and strong mechanical properties are desired for smart materials used in soft actuators. A bioinspired hybrid hydrogel actuator was designed and prepared by series combination of three trunks of tough polymer-clay hydrogels to accomplish the comprehensive actuation of "extension-grasp-retraction" like a fishing rod. The hydrogels with thermo-creep and thermo-shrinking features were successively irradiated by near-infrared (NIR) to execute extension and retraction, respectively. The GO in the hydrogels absorbed the NIR energy and transformed it into thermo-energy rapidly and effectively. The hydrogel with adhesion or magnetic force was adopted as the "hook" of the hybrid hydrogel actuator for grasping object. The hook of the hybrid hydrogel actuator was replaceable according to applications, even with functional materials other than hydrogels. This study provides an innovative concept to explore new soft actuators through combining response hydrogels and programming the same stimulus.

  20. Massachusetts Recreational Fishing Demand Survey

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Stated preference choice experiment data were collected in 2012 from Massachuestts saltwater recreational fishermen. Saltwater anglers fishing in Massachusetts (MA)...

  1. Fish-robot interactions in a free-swimming environment: Effects of speed and configuration of robots on live fish

    Science.gov (United States)

    Butail, Sachit; Polverino, Giovanni; Phamduy, Paul; Del Sette, Fausto; Porfiri, Maurizio

    2014-03-01

    We explore fish-robot interactions in a comprehensive set of experiments designed to highlight the effects of speed and configuration of bioinspired robots on live zebrafish. The robot design and movement is inspired by salient features of attraction in zebrafish and includes enhanced coloration, aspect ratio of a fertile female, and carangiform/subcarangiformlocomotion. The robots are autonomously controlled to swim in circular trajectories in the presence of live fish. Our results indicate that robot configuration significantly affects both the fish distance to the robots and the time spent near them.

  2. Effects of fish oil type, lipid antioxidants and presence of rapeseed oil on oxidative flavour stability of fish oil enriched milk

    DEFF Research Database (Denmark)

    Bruni Let, Mette; Jacobsen, Charlotte; Meyer, Anne S.

    2004-01-01

    As a part of our ongoing experiments on optimization of the oxidative stability of fish oils in genuine food systems, this study investigated the oxidative deterioration of fish oil enriched milk emulsions during cold storage. The experimental data showed that addition of rapeseed oil to fish oil...... (1:1) prior to emulsification into milk significantly protected the emulsions against oxidative deterioration. Addition of propyl gallate and a citric acid ester to the fish oil prior to emulsification also protected the fish oil enriched milk during storage. Emulsions containing a rapeseed:fish oil...... mixture were oxidatively stable during 11 d at 2 øC. Thus, no additional inhibitory effect of the added antioxidants was observed. The peroxide value and concentrations of five selected volatiles derived from n- 3 PUFA degradation in rapeseed:fish oil mixture emulsions were not significantly different...

  3. 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...... biochemical indicators of spoilage. Shewanzella putrefaciens and Pseudomonas spp. are the specific spoilage bacteria of iced fresh fish regardless of the origin of the fish. Modified atmosphere stored marine fish from temperate waters are spoiled by the CO2 resistant Photobacterium phosphoreum whereas Gram......- positive bacteria are likely spoilers of CO2 packed fish from fresh or tropical waters. Fish products with high salt contents may spoil due to growth of halophilic bacteria (salted fish) or growth of anaerobic bacteria and yeasts (barrel salted fish). Whilst the spoilage of fresh and highly salted fish...

  4. Will Tidal Wetland Restoration Enhance Populations of Native Fishes?

    Directory of Open Access Journals (Sweden)

    Larry R. Brown

    2003-10-01

    Full Text Available Restoration of tidal wetlands might enhance populations of native fishes in the San Francisco Estuary of California. The purpose of this paper is to: (1 review the currently available information regarding the importance of tidal wetlands to native fishes in the San Francisco Estuary, (2 construct conceptual models on the basis of available information, (3 identify key areas of scientific uncertainty, and (4 identify methods to improve conceptual models and reduce uncertainty. There are few quantitative data to suggest that restoration of tidal wetlands will substantially increase populations of native fishes. On a qualitative basis, there is some support for the idea that tidal wetland restoration will increase populations of some native fishes; however, the species deriving the most benefit from restoration might not be of great management concern at present. Invasion of the San Francisco Estuary by alien plants and animals appears to be a major factor in obscuring the expected link between tidal wetlands and native fishes. Large-scale adaptive management experiments (>100 hectares appear to be the best available option for determining whether tidal wetlands will provide significant benefit to native fishes. Even if these experiments are unsuccessful at increasing native fish populations, the restored wetlands should benefit native birds, plants, and other organisms.

  5. Reef fish communities in the central Red Sea show evidence of asymmetrical fishing pressure

    KAUST Repository

    Kattan, Alexander; Coker, Darren James; Berumen, Michael L.

    2017-01-01

    In order to assess human impacts and develop rational restoration goals for corals reefs, baseline estimates of fish communities are required. In Saudi Arabian waters of the Red Sea, widespread unregulated fishing is thought to have been ongoing for decades, but there is little direct evidence of the impact on reef communities. To contextualize this human influence, reef-associated fish assemblages on offshore reefs in Saudi Arabia and Sudan in the central Red Sea were investigated. These reefs have comparable benthic environments, experience similar oceanographic influences, and are separated by less than 300 km, offering an ideal comparison for identifying potential anthropogenic impacts such as fishing pressure. This is the first study to assess reef fish biomass in both these regions, providing important baselines estimates. We found that biomass of top predators on offshore Sudanese reefs was on average almost three times that measured on comparable reefs in Saudi Arabia. Biomass values from some of the most remote reefs surveyed in Sudan’s far southern region even approach those previously reported in the Northwestern Hawaiian Islands, northern Line Islands, Pitcairn Islands, and other isolated Pacific islands and atolls. The findings suggest that fishing pressure has significantly altered the fish community structure of Saudi Arabian Red Sea reefs, most conspicuously in the form of top predator removal. The results point towards the urgent need for enhanced regulation and enforcement of fishing practices in Saudi Arabia, while making a strong case for protection in the form of no-take marine protected areas to maintain preservation of the relatively intact southern Sudanese Red Sea.

  6. Reef fish communities in the central Red Sea show evidence of asymmetrical fishing pressure

    KAUST Repository

    Kattan, Alexander

    2017-03-09

    In order to assess human impacts and develop rational restoration goals for corals reefs, baseline estimates of fish communities are required. In Saudi Arabian waters of the Red Sea, widespread unregulated fishing is thought to have been ongoing for decades, but there is little direct evidence of the impact on reef communities. To contextualize this human influence, reef-associated fish assemblages on offshore reefs in Saudi Arabia and Sudan in the central Red Sea were investigated. These reefs have comparable benthic environments, experience similar oceanographic influences, and are separated by less than 300 km, offering an ideal comparison for identifying potential anthropogenic impacts such as fishing pressure. This is the first study to assess reef fish biomass in both these regions, providing important baselines estimates. We found that biomass of top predators on offshore Sudanese reefs was on average almost three times that measured on comparable reefs in Saudi Arabia. Biomass values from some of the most remote reefs surveyed in Sudan’s far southern region even approach those previously reported in the Northwestern Hawaiian Islands, northern Line Islands, Pitcairn Islands, and other isolated Pacific islands and atolls. The findings suggest that fishing pressure has significantly altered the fish community structure of Saudi Arabian Red Sea reefs, most conspicuously in the form of top predator removal. The results point towards the urgent need for enhanced regulation and enforcement of fishing practices in Saudi Arabia, while making a strong case for protection in the form of no-take marine protected areas to maintain preservation of the relatively intact southern Sudanese Red Sea.

  7. Irradiated Palm Oil Waste (Sludge) As Feed Supplement For Nila Gift Fish (Oreochromis niloticus)

    International Nuclear Information System (INIS)

    MU, Jenny; PM, Adria

    2002-01-01

    The objective of the experiment was to study the fish weight development after being fed with irradiated palm oil waste pellet. Irradiated Palm oil waste pellet was produced from palm oil waste (sludge) with some additional materials, i.e. rice bran, fish powder, soybean powder, tapioca powder. The mixture was then irradiated with a dose of 4 kGy to decontaminate pathogen microbe and other contaminant microbes, the experiment have been carried out in 4 treatments. Treatment A was male fish which was being fed with irradiated sludge palm oil waste pellet and commercial pelletized feed (2:1), treatment C was female with the same feed as A, treatment B was male fish feed with commercial pelletized, treatment D was female fish with the same feed as B. Each treatment was placed in a pond. The feed with the amount of 3% of total body weight was given to the fishes 2 times per day. The result of this experiment showed that the male fish weight receiving treatment A and B were 195.37 g and 175.12 g. The female fish weight at treatments C and D were 170.28 g and 160.15 g, respectively. Data obtained from this experiment showed that the treatment of irradiated sludge palm oil waste pellet and commercial pelletized (2:1) were more efficient as fish feeding compared to commercial pellets

  8. Resolving colocalization of bacteria and metal(loid)s on plant root surfaces by combining fluorescence in situ hybridization (FISH) with multiple-energy micro-focused X-ray fluorescence (ME μXRF).

    Science.gov (United States)

    Honeker, Linnea K; Root, Robert A; Chorover, Jon; Maier, Raina M

    2016-12-01

    Metal(loid)-contamination of the environment due to anthropogenic activities is a global problem. Understanding the fate of contaminants requires elucidation of biotic and abiotic factors that influence metal(loid) speciation from molecular to field scales. Improved methods are needed to assess micro-scale processes, such as those occurring at biogeochemical interfaces between plant tissues, microbial cells, and metal(loid)s. Here we present an advanced method that combines fluorescence in situ hybridization (FISH) with synchrotron-based multiple-energy micro-focused X-ray fluorescence microprobe imaging (ME μXRF) to examine colocalization of bacteria and metal(loid)s on root surfaces of plants used to phytostabilize metalliferous mine tailings. Bacteria were visualized on a small root section using SytoBC nucleic acid stain and FISH probes targeting the domain Bacteria and a specific group (Alphaproteobacteria, Gammaproteobacteria, or Actinobacteria). The same root region was then analyzed for elemental distribution and metal(loid) speciation of As and Fe using ME μXRF. The FISH and ME μXRF images were aligned using ImageJ software to correlate microbiological and geochemical results. Results from quantitative analysis of colocalization show a significantly higher fraction of As colocalized with Fe-oxide plaques on the root surfaces (fraction of overlap 0.49±0.19) than to bacteria (0.072±0.052) (proots, metal(loid)s and microbes, information that should lead to improved mechanistic models of metal(loid) speciation and fate. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Effects of contrasting omnivorous fish on submerged macrophyte biomass in temperate lakes: a mesocosm experiment

    NARCIS (Netherlands)

    Dorenbosch, M.; Bakker, E.S.

    2012-01-01

    1.Freshwater fish can affect aquatic vegetation directly by consuming macrophytes or indirectly by changing water quality. However, most fish in the temperate climate zone have an omnivorous diet. The impact of fish as aquatic herbivores in temperate climates therefore remains unclear and depends on

  10. Design and initial operation of a two-color soft x-ray camera system on the Compact Toroidal Hybrid experiment

    International Nuclear Information System (INIS)

    Herfindal, J. L.; Dawson, J. D.; Ennis, D. A.; Hartwell, G. J.; Loch, S. D.; Maurer, D. A.

    2014-01-01

    A multi-camera soft x-ray diagnostic has been developed to measure the equilibrium electron temperature profile and temperature fluctuations due to magnetohydrodynamic activity on the Compact Toroidal Hybrid experiment. The diagnostic consists of three separate cameras each employing two 20-channel diode arrays that view the same plasma region through different beryllium filter thicknesses of 1.8 μm and 3.0 μm allowing electron temperature measurements between 50 eV and 200 eV. The Compact Toroidal Hybrid is a five-field period current-carrying stellarator, in which the presence of plasma current strongly modifies the rotational transform and degree of asymmetry of the equilibrium. Details of the soft x-ray emission, effects of plasma asymmetry, and impurity line radiation on the design and measurement of the two-color diagnostic are discussed. Preliminary estimates of the temperature perturbation due to sawtooth oscillations observed in these hybrid discharges are given

  11. Repetitive sequences: the hidden diversity of heterochromatin in prochilodontid fish

    Directory of Open Access Journals (Sweden)

    Maria L. Terencio

    2015-08-01

    Full Text Available The structure and organization of repetitive elements in fish genomes are still relatively poorly understood, although most of these elements are believed to be located in heterochromatic regions. Repetitive elements are considered essential in evolutionary processes as hotspots for mutations and chromosomal rearrangements, among other functions – thus providing new genomic alternatives and regulatory sites for gene expression. The present study sought to characterize repetitive DNA sequences in the genomes of Semaprochilodus insignis (Jardine & Schomburgk, 1841 and Semaprochilodus taeniurus (Valenciennes, 1817 and identify regions of conserved syntenic blocks in this genome fraction of three species of Prochilodontidae (S. insignis, S. taeniurus, and Prochilodus lineatus (Valenciennes, 1836 by cross-FISH using Cot-1 DNA (renaturation kinetics probes. We found that the repetitive fractions of the genomes of S. insignis and S. taeniurus have significant amounts of conserved syntenic blocks in hybridization sites, but with low degrees of similarity between them and the genome of P. lineatus, especially in relation to B chromosomes. The cloning and sequencing of the repetitive genomic elements of S. insignis and S. taeniurus using Cot-1 DNA identified 48 fragments that displayed high similarity with repetitive sequences deposited in public DNA databases and classified as microsatellites, transposons, and retrotransposons. The repetitive fractions of the S. insignis and S. taeniurus genomes exhibited high degrees of conserved syntenic blocks in terms of both the structures and locations of hybridization sites, but a low degree of similarity with the syntenic blocks of the P. lineatus genome. Future comparative analyses of other prochilodontidae species will be needed to advance our understanding of the organization and evolution of the genomes in this group of fish.

  12. Laboratory Experiments on the Effects of Blade Strike from Hydrokinetic Energy Technologies on Larval and Juvenile Freshwater Fishes

    Energy Technology Data Exchange (ETDEWEB)

    Schweizer, Peter E [ORNL; Cada, Glenn F [ORNL; Bevelhimer, Mark S [ORNL

    2012-03-01

    concern that small, fragile fish early life stages may be unable to avoid being struck by the blades of hydrokinetic turbines, we found no empirical data in the published literature that document survival of earliest life-stage fish in passage by rotor blades. In addition to blade strike, research on passage of fish through conventional hydropower turbines suggested that fish mortalities from passage through the rotor swept area could also occur due to shear stresses and pressure chances in the water column (Cada et al. 1997, Turnpenny 1998). However, for most of the proposed HK turbine designs the rotors are projected to operate a lower RPM (revolutions per minute) than observed from conventional reaction turbines; the associated shear stress and pressure changes are expected to be lower and pose a smaller threat to fish survival (DOE 2009). Only a limited number of studies have been conducted to examine the risk of blade strike from hydrokinetic technologies to fish (Turnpenny et al. 1992, Normandeau et al. 2009, Seitz et al. 2011, EPRI 2011); the survival of drifting or weakly swimming fish (especially early life stages) that encounter rotor blades from hydrokinetic (HK) devices is currently unknown. Our study addressed this knowledge gap by testing how fish larvae and juveniles encountered different blade profiles of hydrokinetic devices and how such encounters influenced survivorship. We carried out a laboratory study designed to improve our understanding of how fish larvae and juvenile fish may be affected by encounters with rotor blades from HK turbines in the water column of river and ocean currents. (For convenience, these early life stages will be referred to as young of the year, YOY). The experiments developed information needed to quantify the risk (both probability and consequences) of rotor-blade strike to YOY fish. In particular, this study attempted to determine whether YOY drifting in a high-velocity flow directly in the path of the blade leading edge

  13. Microbiological spoilage of fish and fish products.

    Science.gov (United States)

    Gram, L; Huss, H H

    1996-11-01

    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 biochemical indicators of spoilage. Shewanella putrefaciens and Pseudomonas spp. are the specific spoilage bacteria of iced fresh fish regardless of the origin of the fish. Modified atmosphere stored marine fish from temperate waters are spoiled by the CO2 resistant Photobacterium phosphoreum whereas Gram-positive bacteria are likely spoilers of CO2 packed fish from fresh or tropical waters. Fish products with high salt contents may spoil due to growth of halophilic bacteria (salted fish) or growth of anaerobic bacteria and yeasts (barrel salted fish). Whilst the spoilage of fresh and highly salted fish is well understood, much less is known about spoilage of lightly preserved fish products. It is concluded that the spoilage is probably caused by lactic acid bacteria, certain psychotrophic Enterobacteriaceae and/or Photobacterium phosphoreum. However, more work is needed in this area.

  14. Investigating the Inverse Square Law with the Timepix Hybrid Silicon Pixel Detector: A CERN [at] School Demonstration Experiment

    Science.gov (United States)

    Whyntie, T.; Parker, B.

    2013-01-01

    The Timepix hybrid silicon pixel detector has been used to investigate the inverse square law of radiation from a point source as a demonstration of the CERN [at] school detector kit capabilities. The experiment described uses a Timepix detector to detect the gamma rays emitted by an [superscript 241]Am radioactive source at a number of different…

  15. Growth of plume ''resident'' fishes in Lake Michigan

    International Nuclear Information System (INIS)

    Spigarelli, S.A.; Smith, D.W.

    1974-01-01

    Brown trout, rainbow trout, and chinook salmon were collected from the Point Beach thermal discharge area, tagged with commercial dart tags and temperature-sensitive tags, and released back into the discharge area. RNA and DNA analyses were performed on epaxial muscle samples taken from each tagged fish recaptured in the plume area and from control fish. A table is presented to show mean weights, condition factors, and RNA-DNA ratios for each group of fish. Results indicated that the fish did not experience any severe growth abnormalities as a result of their residence in the thermal plume area

  16. Invasive hybridization in a threatened species is accelerated by climate change

    Science.gov (United States)

    Muhlfeld, Clint C.; Kovach, Ryan P.; Jones, Leslie A.; Al-Chokhachy, Robert K.; Boyer, Matthew C.; Leary, Robb F.; Lowe, Winsor H.; Luikart, Gordon; Allendorf, Fred W.

    2014-01-01

    Climate change will decrease worldwide biodiversity through a number of potential pathways, including invasive hybridization (cross-breeding between invasive and native species). How climate warming influences the spread of hybridization and loss of native genomes poses difficult ecological and evolutionary questions with little empirical information to guide conservation management decisions. Here we combine long-term genetic monitoring data with high-resolution climate and stream temperature predictions to evaluate how recent climate warming has influenced the spatio-temporal spread of human-mediated hybridization between threatened native westslope cutthroat trout (Oncorhynchus clarkii lewisi) and non-native rainbow trout (Oncorhynchus mykiss), the world’s most widely introduced invasive fish. Despite widespread release of millions of rainbow trout over the past century within the Flathead River system, a large relatively pristine watershed in western North America, historical samples revealed that hybridization was prevalent only in one (source) population. During a subsequent 30-year period of accelerated warming, hybridization spread rapidly and was strongly linked to interactions between climatic drivers—precipitation and temperature—and distance to the source population. Specifically, decreases in spring precipitation and increases in summer stream temperature probably promoted upstream expansion of hybridization throughout the system. This study shows that rapid climate warming can exacerbate interactions between native and non-native species through invasive hybridization, which could spell genomic extinction for many species.

  17. Invasive hybridization in a threatened species is accelerated by climate change

    Science.gov (United States)

    Muhlfeld, Clint C.; Kovach, Ryan P.; Jones, Leslie A.; Al-Chokhachy, Robert; Boyer, Matthew C.; Leary, Robb F.; Lowe, Winsor H.; Luikart, Gordon; Allendorf, Fred W.

    2014-07-01

    Climate change will decrease worldwide biodiversity through a number of potential pathways, including invasive hybridization (cross-breeding between invasive and native species). How climate warming influences the spread of hybridization and loss of native genomes poses difficult ecological and evolutionary questions with little empirical information to guide conservation management decisions. Here we combine long-term genetic monitoring data with high-resolution climate and stream temperature predictions to evaluate how recent climate warming has influenced the spatio-temporal spread of human-mediated hybridization between threatened native westslope cutthroat trout (Oncorhynchus clarkii lewisi) and non-native rainbow trout (Oncorhynchus mykiss), the world's most widely introduced invasive fish. Despite widespread release of millions of rainbow trout over the past century within the Flathead River system, a large relatively pristine watershed in western North America, historical samples revealed that hybridization was prevalent only in one (source) population. During a subsequent 30-year period of accelerated warming, hybridization spread rapidly and was strongly linked to interactions between climatic drivers--precipitation and temperature--and distance to the source population. Specifically, decreases in spring precipitation and increases in summer stream temperature probably promoted upstream expansion of hybridization throughout the system. This study shows that rapid climate warming can exacerbate interactions between native and non-native species through invasive hybridization, which could spell genomic extinction for many species.

  18. Combination of microautoradiography and fluorescence in situ hybridization for identification of microorganisms degrading xenobiotic contaminants.

    Science.gov (United States)

    Yang, Yanru; Zarda, Annatina; Zeyer, Josef

    2003-12-01

    One of the central topics in environmental bioremediation research is to identify microorganisms that are capable of degrading the contaminants of interest. Here we report application of combined microautoradiography (MAR) and fluorescence in situ hybridization (FISH). The method has previously been used in a number of systems; however, here we demonstrate its feasibility in studying the degradation of xenobiotic compounds. With a model system (coculture of Pseudomonas putida B2 and Sphingomonas stygia incubated with [14C] o-nitrophenol), combination of MAR and FISH was shown to be able to successfully identify the microorganisms degrading o-nitrophenol. Compared with the conventional techniques, MAR-FISH allows fast and accurate identification of the microorganisms involved in environmental contaminant degradation.

  19. Fluorescence in situ hybridization in combination with the comet assay and micronucleus test in genetic toxicology

    Directory of Open Access Journals (Sweden)

    Hovhannisyan Galina G

    2010-09-01

    Full Text Available Abstract Comet assay and micronucleus (MN test are widely applied in genotoxicity testing and biomonitoring. While comet assay permits to measure direct DNA-strand breaking capacity of a tested agent MN test allows estimating the induced amount of chromosome and/or genome mutations. The potential of these two methods can be enhanced by the combination with fluorescence in situ hybridization (FISH techniques. FISH plus comet assay allows the recognition of targets of DNA damage and repairing directly. FISH combined with MN test is able to characterize the occurrence of different chromosomes in MN and to identify potential chromosomal targets of mutagenic substances. Thus, combination of FISH with the comet assay or MN test proved to be promising techniques for evaluation of the distribution of DNA and chromosome damage in the entire genome of individual cells. FISH technique also permits to study comet and MN formation, necessary for correct application of these methods. This paper reviews the relevant literature on advantages and limitations of Comet-FISH and MN-FISH assays application in genetic toxicology.

  20. Muscle dynamics in fish during steady swimming

    DEFF Research Database (Denmark)

    Shadwick, RE; Steffensen, JF; Katz, SL

    1998-01-01

    SYNOPSIS. Recent research in fish locomotion has been dominated by an interest in the dynamic mechanical properties of the swimming musculature. Prior observations have indicated that waves of muscle activation travel along the body of an undulating fish faster than the resulting waves of muscular...... position in swimming fish. Quantification of muscle contractile properties in cyclic contractions relies on in vitro experiments using strain and activation data collected in vivo. In this paper we discuss the relation between these parameters and body kinematics. Using videoradiographic data from swimming...... constant cross-section of red muscle along much of the body suggests that positive power for swimming is generated fairly uniformly along the length of the fish....

  1. TA3 - Dosimetry and instrumentation supply of the M-Fish technique to the Fish-3 painting technique for analysing translocations: A radiotherapy-treated patient study

    International Nuclear Information System (INIS)

    Pouzoulet, F.; Roch-Lefevre, S.; Giraudet, A.L.; Vaurijoux, A.; Voisin, P.A.; Buard, V.; Delbos, M.; Voisin, Ph.; Roy, L.; Bourhis, J.

    2006-01-01

    Purpose: Currently, the chromosome translocation study is the best method to estimate the dose of an old radiation exposure. Fluorescent In Situ Hybridization (F.I.S.H.) technique allows an easy detection of this kind of aberrations. However, as only a few number of chromosomes is usually painted, some bias could skew the result. To evaluate the advantage of using full genome staining (M-F.I.S.H. technique) compared with three chromosomes labelling (F.I.S.H.-3 painting), we compared translocation yields in radiotherapy treated patients. Methods: Chromosome aberration analyses were performed on peripheral blood lymphocyte cultures of two patients treated for a throat cancer by radiotherapy. Blood samples were obtained, before, along the treatment and six or four months later. For each sample, a dicentrics analysis was performed together with translocation analysis either with F.I.S.H.-3 painting or M-F.I.S.H.. Results: By confronting results from the F.I.S.H.-3 painting technique and the M-F.I.S.H. technique, significant differences were revealed. The translocations yield seemed to be stable with the F.I.S.H.-3 painting technique whereas it is not the case with the M-F.I.S.H. technique. This difference in results was explained by the bias induced by F.I.S.H.-3 Painting technique in the visualisation of complex aberrations. Furthermore, we found the presence of a clone bearing a translocation involving a painted chromosome. Conclusions: According to the potential bias of F.I.S.H.-3 painting on translocations study, the M-F.I.S.H. technique should provide more precise and reproducible results. Because of its more difficult implement, it seems hardly applicable to retrospective dosimetry instead of F.I.S.H.-3 painting technique. (authors)

  2. TA3 - Dosimetry and instrumentation supply of the M-Fish technique to the Fish-3 painting technique for analysing translocations: A radiotherapy-treated patient study

    Energy Technology Data Exchange (ETDEWEB)

    Pouzoulet, F.; Roch-Lefevre, S.; Giraudet, A.L.; Vaurijoux, A.; Voisin, P.A.; Buard, V.; Delbos, M.; Voisin, Ph.; Roy, L. [Institut de Radioprotection et de Surete Nucleaire, Lab. de Dosimetrie Biologique, 92 - Fontenay aux Roses (France); Bourhis, J. [Laboratoire UPRES EA 27-10, Radiosensibilite des Tumeurs et Tissus sains, PR1, 94 - Villejuif (France)

    2006-07-01

    Purpose: Currently, the chromosome translocation study is the best method to estimate the dose of an old radiation exposure. Fluorescent In Situ Hybridization (F.I.S.H.) technique allows an easy detection of this kind of aberrations. However, as only a few number of chromosomes is usually painted, some bias could skew the result. To evaluate the advantage of using full genome staining (M-F.I.S.H. technique) compared with three chromosomes labelling (F.I.S.H.-3 painting), we compared translocation yields in radiotherapy treated patients. Methods: Chromosome aberration analyses were performed on peripheral blood lymphocyte cultures of two patients treated for a throat cancer by radiotherapy. Blood samples were obtained, before, along the treatment and six or four months later. For each sample, a dicentrics analysis was performed together with translocation analysis either with F.I.S.H.-3 painting or M-F.I.S.H.. Results: By confronting results from the F.I.S.H.-3 painting technique and the M-F.I.S.H. technique, significant differences were revealed. The translocations yield seemed to be stable with the F.I.S.H.-3 painting technique whereas it is not the case with the M-F.I.S.H. technique. This difference in results was explained by the bias induced by F.I.S.H.-3 Painting technique in the visualisation of complex aberrations. Furthermore, we found the presence of a clone bearing a translocation involving a painted chromosome. Conclusions: According to the potential bias of F.I.S.H.-3 painting on translocations study, the M-F.I.S.H. technique should provide more precise and reproducible results. Because of its more difficult implement, it seems hardly applicable to retrospective dosimetry instead of F.I.S.H.-3 painting technique. (authors)

  3. Organization and variation analysis of 5S rDNA in different ploidy-level hybrids of red crucian carp × topmouth culter.

    Science.gov (United States)

    He, Weiguo; Qin, Qinbo; Liu, Shaojun; Li, Tangluo; Wang, Jing; Xiao, Jun; Xie, Lihua; Zhang, Chun; Liu, Yun

    2012-01-01

    Through distant crossing, diploid, triploid and tetraploid hybrids of red crucian carp (Carassius auratus red var., RCC♀, Cyprininae, 2n = 100) × topmouth culter (Erythroculter ilishaeformis Bleeker, TC♂, Cultrinae, 2n = 48) were successfully produced. Diploid hybrids possessed 74 chromosomes with one set from RCC and one set from TC; triploid hybrids harbored 124 chromosomes with two sets from RCC and one set from TC; tetraploid hybrids had 148 chromosomes with two sets from RCC and two sets from TC. The 5S rDNA of the three different ploidy-level hybrids and their parents were sequenced and analyzed. There were three monomeric 5S rDNA classes (designated class I: 203 bp; class II: 340 bp; and class III: 477 bp) in RCC and two monomeric 5S rDNA classes (designated class IV: 188 bp, and class V: 286 bp) in TC. In the hybrid offspring, diploid hybrids inherited three 5S rDNA classes from their female parent (RCC) and only class IV from their male parent (TC). Triploid hybrids inherited class II and class III from their female parent (RCC) and class IV from their male parent (TC). Tetraploid hybrids gained class II and class III from their female parent (RCC), and generated a new 5S rDNA sequence (designated class I-N). The specific paternal 5S rDNA sequence of class V was not found in the hybrid offspring. Sequence analysis of 5S rDNA revealed the influence of hybridization and polyploidization on the organization and variation of 5S rDNA in fish. This is the first report on the coexistence in vertebrates of viable diploid, triploid and tetraploid hybrids produced by crossing parents with different chromosome numbers, and these new hybrids are novel specimens for studying the genomic variation in the first generation of interspecific hybrids, which has significance for evolution and fish genetics.

  4. Luciferase genes cloned from the unculturable luminous bacteroid symbiont of the Caribbean flashlight fish, Kryptophanaron alfredi.

    Science.gov (United States)

    Haygood, M G; Cohn, D H

    1986-01-01

    Light organs of anomalopid (flashlight) fish contain luminous bacteroids that have never been cultured and, consequently, have been difficult to study. We have characterized the luciferase (lux) region of DNA extracted from light organs of the Caribbean flashlight fish Kryptophanaron alfredi by hybridization of cloned Vibrio harveyi lux genes to restriction-endonuclease-digested, light organ DNA. Comparison of the hybridization pattern of light organ DNA with that of DNA of a putative symbiotic isolate provides a method for identifying the authentic luminous symbiont regardless of its luminescence, and was used to reject one such isolate. Light organ DNA was further used to construct a cosmid clone bank and the luciferase genes were isolated. Unlike other bacterial luciferase genes, the genes were not expressed in Escherichia coli. When placed under the control of the E. coli trp promoter, the genes were transcribed but no luciferase was detected, suggesting a posttranscriptional block to expression.

  5. Fish Swimming and Bird/Insect Flight

    Science.gov (United States)

    Wu, Theodore Yaotsu

    2011-01-01

    This expository review is devoted to fish swimming and bird/insect flight. (a) The simple waving motion of an elongated flexible ribbon plate of constant width propagating a wave distally down the plate to swim forward in a fluid, initially at rest, is first considered to provide a fundamental concept on energy conservation. It is generalized to include variations in body width and thickness, with appended dorsal, ventral and caudal fins shedding vortices to closely simulate fish swimming, for which a nonlinear theory is presented for large-amplitude propulsion. (b) For bird flight, the pioneering studies on oscillatory rigid wings are discussed with delineating a fully nonlinear unsteady theory for a two-dimensional flexible wing with arbitrary variations in shape and trajectory to provide a comparative study with experiments. (c) For insect flight, recent advances are reviewed by items on aerodynamic theory and modeling, computational methods, and experiments, for forward and hovering flights with producing leading-edge vortex to yield unsteady high lift. (d) Prospects are explored on extracting prevailing intrinsic flow energy by fish and bird to enhance thrust for propulsion. (e) The mechanical and biological principles are drawn together for unified studies on the energetics in deriving metabolic power for animal locomotion, leading to the surprising discovery that the hydrodynamic viscous drag on swimming fish is largely associated with laminar boundary layers, thus drawing valid and sound evidences for a resounding resolution to the long-standing fish-swim paradox proclaimed by Gray (1936, 1968 ).

  6. Fluorescence in situ hybridization for phytoplasma and endophytic bacteria localization in plant tissues.

    Science.gov (United States)

    Bulgari, Daniela; Casati, Paola; Faoro, Franco

    2011-11-01

    In the present study, we developed a rapid and efficient fluorescence in situ hybridization assay (FISH) in non-embedded tissues of the model plant Catharanthus roseus for co-localizing phytoplasmas and endophytic bacteria, opening new perspectives for studying the interaction between these microorganisms. Copyright © 2011 Elsevier B.V. All rights reserved.

  7. A Preliminary Assessment of Barotrauma Injuries and Acclimation Studies for Three Fish Species

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Richard S. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Walker, Ricardo W. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Stephenson, John R. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2015-12-15

    Fish that pass hydro structures either through turbine passage, deep spill, or other deep pathways can experience rapid decreases in pressure that can result in barotrauma. In addition to morphology and physiology of the fish’s swim bladder, the severity of barotrauma is directly related to the volume of undissolved gas in fish prior to rapid decompression and the lowest pressure the fish experience as they pass hydro structures (termed the “nadir”). The volume of undissolved gas in fish is influenced by the depth of acclimation (the pressure at which the fish is neutrally buoyant); therefore, determining the depth where fish are neutrally buoyant is a critical precursor to determining the relationship between pressure changes and injury or mortality.

  8. Laboratory studies on the effects of shear on fish: Final report

    Energy Technology Data Exchange (ETDEWEB)

    Neitzel, Duane A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Richmond, M. C. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Dauble, D. D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Mueller, R. P. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Moursund, R. A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Abernethy, C. S. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guensch, G. R. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Cada, G. F. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2000-09-01

    The overall objective of these studies was to specify an index describing the hydraulic force that fish experience when subjected to a shear environment. Fluid shear is a phenomenon that is important to fish. However, elevated levels of shear may result in strain rates that injure or kill fish. At hydroelectric generating facilities, concerns have been expressed that strain rates associated with passage through turbines, spillways, and fish bypass systems may adversely affect migrating fish. Development of fish-friendly hydroelectric turbines requires knowledge of the physical forces (injury mechanisms) that impact entrained fish and the fish’s tolerance to these forces. It requires up-front, pre-design specifications for the environmental conditions that occur within the turbine system; in other words, determining or assuming conditions known to injure fish will assist engineers in the design of a fish-friendly turbine system. To address the development of biological specifications, this experiment designed and built a test facility where juvenile fish could be subjected to a range of shear environments and quantified their biological response. The test data reported here provide quantified strain rates and the relationship of these forces to direct and indirect biological effects on fish. The study concludes that juvenile salmonids and American shad should survive shear environments where strain rates do not exceed 500 cm/s/cm at a Dy of 1.8 cm. Additional studies are planned with a sensor fish to better link hydraulic conditions found within the laboratory and field environments.

  9. Cosmic ray nuclear interactions and EAS-triggered families observed by the Chacaltaya hybrid experiment

    International Nuclear Information System (INIS)

    Aoki, H.; Honda, K.; Inoue, N.; Ishii, T.; Kawasumi, N.; Martinic, N.; Ochi, N.; Ohmori, N.; Ohsawa, A.; Tamada, M.; Ticona, R.

    2008-01-01

    Longitudinal and lateral characteristics of the families detected by emulsion chambers in the hybrid experiment with AS-array at Mt. Chacaltaya are studied in detail. Although many groups discuss about an increase of the heavy component in primary cosmic-rays beyond the knee region, it is shown that the observed characteristics of the families accompanied by large shower size, Ne>10 6 , can not be explained by an increase of heavy primaries alone. It is necessary to assume some changes of nuclear interaction in order to explain the observed characteristics of the air-showers accompanied by families

  10. Microbial abundance and diversity in water, and immune parameters of red tilapia reared in bioflocs system with different fish density (25 fish/m3, 50 fish/m3, and 100 fish/m3

    Directory of Open Access Journals (Sweden)

    Frid Agustinus

    2010-07-01

    Full Text Available ABSTRACTThe objective of this experiment was to study microbial abundance and diversity in the water, and immune parameters of red tilapia Oreochromis sp. cultured in bioflok system with different fish stocking densities. The experiment comprised of two different factors, carbon source addition (bioflocs and control, and fish stocking density (25 fish/m3, 50 fish/m3, dan 100 fish/m3, with an experimental period of 99 days. Microbial load in water was determined biweekly, whereas immune parameters represented by fish blood profile were measured on day 0, 50, and 90. There was no significant difference in total bacteria count in the water of all treatments; there was however a tendency shown by all treatments that the microbial load in water increased along with the culture period. There were 4 genera of bacteria which particularly found in bioflok system, which are Acinetobacter sp., Corynobacterium sp., Listeria sp., dan Pseudomonas sp, and are suggested to play a role in bioflok formation. The percentage of phagocytic index of fish in bioflok system was higher than that in control, and may indicate that bioflok may stimulate the fish immune system.Keywords: bioflocs, red tilapia, bacteria, blood profile. ABSTRAKPenelitian ini bertujuan untuk mengidentifikasi kelimpahan dan keragaman jenis bakteri dalam air dan parameter imunitas ikan nila Oreochromis sp. yang dipelihara dalam sistem bioflok dengan kepadatan ikan yang berbeda. Penelitian terdiri atas dua faktor perlakuan yaitu penambahan sumber carbon (bioflok dan kontrol, dan padat penebaran ikan (25 ekor/m3, 50 ekor/m3, dan 100 ekor/m3 dengan lama waktu pemeliharaan ikan selama 99 hari. Kelimpahan bakteri diukur setiap 2 minggu sekali selama masa pemeliharaan. Parameter imunitas meliputi gambaran darah diukur dengan pengambilan contoh darah yang dilakukan pada tiga ekor ikan pada hari ke 0, 50, dan 99. Kelimpahan bakteri pada semua perlakuan pada setiap titik pengamatan tidak menunjukkan

  11. Fish Farm Challenge Provides STEM Design Experiences for Youth

    Science.gov (United States)

    Horton , Robert L.; House, Patty L.

    2015-01-01

    In 2014, Monsanto Corporation partnered with National 4-H Council to help inspire and develop professional skills among young agriculturalists. The Ohio State University created Fish Farm Challenge, which engaged more than 8,000 youth across eight states. Youth were taught about worldwide food insecurity and the importance of aquaculture. They…

  12. Innovation in utilization of fish tanks for fish culture among fish ...

    African Journals Online (AJOL)

    This study investigated innovation in utilization of fish tanks for fish culture among fish farmers in Obio/Akpor Local Government Area of Rivers State, Nigeria. Data for this study was obtained through the administration of questionnaire and scheduled interview to 120 sampled fish farmers randomly selected from the study ...

  13. Fishing Farmers or Farming Fishers? Fishing Typology of Inland Small-Scale Fishing Households and Fisheries Management in Singkarak Lake, West Sumatra, Indonesia

    Science.gov (United States)

    Yuerlita; Perret, Sylvain Roger; Shivakoti, Ganesh P.

    2013-07-01

    Technical and socio-economic characteristics are known to determine different types of fishers and their livelihood strategies. Faced with declining fish and water resources, small-scale fisheries engage into transformations in livelihood and fishing practices. The paper is an attempt to understand these changes and their socio-economic patterns, in the case of Singkarak Lake in West Sumatra, Indonesia. Based upon the hypothesis that riparian communities have diverse, complex yet structured and dynamic livelihood systems, the paper's main objective is to study, document and model the actual diversity in livelihood, practices and performance of inland small-scale fisheries along the Singkarak Lake, to picture how households are adapted to the situation, and propose an updated, workable model (typology) of those for policy. Principal component analysis and cluster analysis were used to develop a typology of fishing households. The results show that small-scale fishers can be classified into different types characterized by distinct livelihood strategies. Three household types are identified, namely "farming fishers" households (type I, 30 %), "fishing farmers" households (type II, 30 %), and "mainly fishers" households (type III, 40 %). There are significant differences among these groups in the number of boats owned, annual fishing income, agriculture income and farming experience. Type I consists of farming fishers, well equipped, with high fishing costs and income, yet with the lowest return on fishing assets. They are also landowners with farming income, showing the lowest return on land capital. Type II includes poor fishing farmers, landowners with higher farming income; they show the highest return on land asset. They have less fishing equipment, costs and income. Type III (mainly fishers) consists of poorer, younger fishers, with highest return on fishing assets and on fishing costs. They have little land, low farming income, and diversified livelihood

  14. 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...

  15. Establishment of a new human pleomorphic malignant fibrous histiocytoma cell line, FU-MFH-2: molecular cytogenetic characterization by multicolor fluorescence in situ hybridization and comparative genomic hybridization

    Directory of Open Access Journals (Sweden)

    Isayama Teruto

    2010-11-01

    Full Text Available Abstract Background Pleomorphic malignant fibrous histiocytoma (MFH is one of the most frequent malignant soft tissue tumors in adults. Despite the considerable amount of research on MFH cell lines, their characterization at a molecular cytogenetic level has not been extensively analyzed. Methods and results We established a new permanent human cell line, FU-MFH-2, from a metastatic pleomorphic MFH of a 72-year-old Japanese man, and applied multicolor fluorescence in situ hybridization (M-FISH, Urovysion™ FISH, and comparative genomic hybridization (CGH for the characterization of chromosomal aberrations. FU-MFH-2 cells were spindle or polygonal in shape with oval nuclei, and were successfully maintained in vitro for over 80 passages. The histological features of heterotransplanted tumors in severe combined immunodeficiency mice were essentially the same as those of the original tumor. Cytogenetic and M-FISH analyses displayed a hypotriploid karyotype with numerous structural aberrations. Urovysion™ FISH revealed a homozygous deletion of the p16INK4A locus on chromosome band 9p21. CGH analysis showed a high-level amplification of 9q31-q34, gains of 1p12-p34.3, 2p21, 2q11.2-q21, 3p, 4p, 6q22-qter, 8p11.2, 8q11.2-q21.1, 9q21-qter, 11q13, 12q24, 15q21-qter, 16p13, 17, 20, and X, and losses of 1q43-qter, 4q32-qter, 5q14-q23, 7q32-qter, 8p21-pter, 8q23, 9p21-pter, 10p11.2-p13, and 10q11.2-q22. Conclusion The FU-MFH-2 cell line will be a particularly useful model for studying molecular pathogenesis of human pleomorphic MFH.

  16. Fluorescence In Situ Hybridization for MicroRNA Detection in Archived Oral Cancer Tissues

    Directory of Open Access Journals (Sweden)

    Zonggao Shi

    2012-01-01

    Full Text Available The noncoding RNA designated as microRNA (miRNA is a large group of small single-stranded regulatory RNA and has generated wide-spread interest in human disease studies. To facilitate delineating the role of microRNAs in cancer pathology, we sought to explore the feasibility of detecting microRNA expression in formalin-fixed paraffin-embedded (FFPE tissues. Using FFPE materials, we have compared fluorescent in situ hybridization (FISH procedures to detect miR-146a with (a different synthetic probes: regular custom DNA oligonucleotides versus locked nucleic acid (LNA incorporated DNA oligonucleotides; (b different reporters for the probes: biotin versus digoxigenin (DIG; (c different visualization: traditional versus tyramide signal amplification (TSA system; (d different blocking reagents for endogenous peroxidase. Finally, we performed miR-146a FISH on a commercially available oral cancer tissue microarray, which contains 40 cases of oral squamous cell carcinoma (OSCC and 10 cases of normal epithelia from the human oral cavity. A sample FISH protocol for detecting miR-146a is provided. In summary, we have established reliable in situ hybridization procedures for detecting the expression of microRNA in FFPE oral cancer tissues. This method is an important tool for studies on the involvement of microRNA in oral cancer pathology and may have potential prognostic or diagnostic value.

  17. Fish oil quality of by-product (fish skin from swangi fish

    Directory of Open Access Journals (Sweden)

    La Ode Huli

    2015-01-01

    Full Text Available The skin of swangi fish is a potential fish skin to be produced for fish oil. The objectives of this research were aimed to determine the yield and the best quality of fish oil and also to compare fatty acid profile of the fish according to different extraction methods. Fish oil extractions were used by wet rendering method with extraction temperatures of 60, 70, 80, 90, 100°C for 20, 30, and 40 minutes. Fish oil quality was determined by the chemical oil characteristics i.e. PV, FFA, AV, anisidin, and TOTOX. Fatty acid profile was analyzed using gas chromatography (Shimadzu. The results of the study showed that the highest fish oil yield in each treatment was obtained extraction temperature of 60°C for 30 minutes with percentage of 0.33, (70°C for 30 minutes 0.46, (80°C for 30 minutes 1.23, (90°C for 20 minutes 1.14 and (100°C for 20 minutes 0.84. These values were lower compare to Bligh & Dyer and Soxhlet methods. Then, the best fish oil quality was resulted on temperature extraction of 60°C for 30 minutes with PV, FFA, anisidin, AV, and TOTOX were 9.17 meq/kg, 6.92%, 13,77 mg KOH/g, 0.86 meq/kg and 19.19 meq/kg, respectively. FUFA fatty acid compositions of swangi skin fish oil especially EPA and DHA in wet rendering method were gained 0.73% and 2.53%, respectively. These results were lower than Bligh & Dyer method which was consisted of 3.66% (EPA, and 13.29% (DHA and also Soxhlet extraction method with value of EPA was 2.78% and DHA was 9.62%.Keywords: EPA, extraction temperature, DHA, fish oil quality, fish skin

  18. Isolation and characterization of 5S rDNA sequences in catfishes genome (Heptapteridae and Pseudopimelodidae): perspectives for rDNA studies in fish by C0t method.

    Science.gov (United States)

    Gouveia, Juceli Gonzalez; Wolf, Ivan Rodrigo; de Moraes-Manécolo, Vivian Patrícia Oliveira; Bardella, Vanessa Belline; Ferracin, Lara Munique; Giuliano-Caetano, Lucia; da Rosa, Renata; Dias, Ana Lúcia

    2016-12-01

    Sequences of 5S ribosomal RNA (rRNA) are extensively used in fish cytogenomic studies, once they have a flexible organization at the chromosomal level, showing inter- and intra-specific variation in number and position in karyotypes. Sequences from the genome of Imparfinis schubarti (Heptapteridae) were isolated, aiming to understand the organization of 5S rDNA families in the fish genome. The isolation of 5S rDNA from the genome of I. schubarti was carried out by reassociation kinetics (C 0 t) and PCR amplification. The obtained sequences were cloned for the construction of a micro-library. The obtained clones were sequenced and hybridized in I. schubarti and Microglanis cottoides (Pseudopimelodidae) for chromosome mapping. An analysis of the sequence alignments with other fish groups was accomplished. Both methods were effective when using 5S rDNA for hybridization in I. schubarti genome. However, the C 0 t method enabled the use of a complete 5S rRNA gene, which was also successful in the hybridization of M. cottoides. Nevertheless, this gene was obtained only partially by PCR. The hybridization results and sequence analyses showed that intact 5S regions are more appropriate for the probe operation, due to conserved structure and motifs. This study contributes to a better understanding of the organization of multigene families in catfish's genomes.

  19. Kalispel Non-Native Fish Suppression Project 2007 Annual Report.

    Energy Technology Data Exchange (ETDEWEB)

    Wingert, Michele; Andersen, Todd [Kalispel Natural Resource Department

    2008-11-18

    Non-native salmonids are impacting native salmonid populations throughout the Pend Oreille Subbasin. Competition, hybridization, and predation by non-native fish have been identified as primary factors in the decline of some native bull trout (Salvelinus confluentus) and westslope cutthroat trout (Oncorhynchus clarki lewisi) populations. In 2007, the Kalispel Natural Resource Department (KNRD) initiated the Kalispel Nonnative Fish Suppression Project. The goal of this project is to implement actions to suppress or eradicate non-native fish in areas where native populations are declining or have been extirpated. These projects have previously been identified as critical to recovering native bull trout and westslope cutthroat trout (WCT). Lower Graham Creek was invaded by non-native rainbow (Oncorhynchus mykiss) and brook trout (Salvelinus fontinalis) after a small dam failed in 1991. By 2003, no genetically pure WCT remained in the lower 700 m of Graham Creek. Further invasion upstream is currently precluded by a relatively short section of steep, cascade-pool stepped channel section that will likely be breached in the near future. In 2008, a fish management structure (barrier) was constructed at the mouth of Graham Creek to preclude further invasion of non-native fish into Graham Creek. The construction of the barrier was preceded by intensive electrofishing in the lower 700 m to remove and relocate all captured fish. Westslope cutthroat trout have recently been extirpated in Cee Cee Ah Creek due to displacement by brook trout. We propose treating Cee Cee Ah Creek with a piscicide to eradicate brook trout. Once eradication is complete, cutthroat trout will be translocated from nearby watersheds. In 2004, the Washington Department of Fish and Wildlife (WDFW) proposed an antimycin treatment within the subbasin; the project encountered significant public opposition and was eventually abandoned. However, over the course of planning this 2004 project, little public

  20. An intergeneric hybrid of a native minnow, the golden shiner, and an exotic minnow, the rudd

    Science.gov (United States)

    Burkhead, N.M.; Williams, J.D.

    1991-01-01

    The hybrid golden shiner Notemigonus crysoleucas × rudd Scardinius erythrophthalmus is the first known nonsalmonid, intergeneric hybrid of an exotic species and a North American native species. The cross is also the first valid record of a viable hybrid involving the native golden shiner. Meristic and mensural characters of 30 artificially produced hybrids of male golden shiners and female rudds were analyzed. Forty-seven percent of the meristic traits exhibited character states intermediate between those of parents. Twenty-seven percent of the meristic characters were supernumerary, suggesting developmental instability of the hybrid genome. Mensural hybrid characters were significantly skewed to the golden shiner phenotype. The skewed mensural inheritance and other skewed patterns of morphological inheritance also suggest problems in canalization of the hybrid phenome or atypical patterns of dominance. All hybrids were identifiable by intermediate squamation of the cultrate abdomen: the keel was mostly scaled but exhibited a small fleshy ridge posteriorly. This minnow hybrid allows general inferences to be made about the phylogenetic affinity of the golden shiner to other cultrate cyprinids of Eurasia. The hybrid cross has important management and conservation implications for fishes in North America. The hybrid is an example of how an exotic species may negatively affect a native species.

  1. Theory of planned behavior applied to fish consumption in modern Metropolitan Lima

    Directory of Open Access Journals (Sweden)

    Angie HIGUCHI

    Full Text Available Abstract Despite being an important source of protein, fish consumption in Peru is low compared with other coastal countries. Thus, the objective of this study is to identify the core determinants of such consumption. We based our analysis on the framework provided by the Theory of Planned Behavior (TPB where attitudes, subjective norms, past experience and health involvement determine the intention and frequency of fish consumption. Primary data were gathered through 159 consumers of fish in modern Metropolitan Lima between August and October 2015. From a set of likert scale indicators a structural model was specified to evaluate the relationships given by the theoretical framework of the TPB. The results showed that the intention to eat fish is determined by personal attitudes, norms and past experience, and as expected, intention itself causes the frequency of fish consumption. Nonetheless, although consumers’ interest in healthy eating was shown to positively influence fish consumption behavior by theory, Metropolitan Lima fish consumers seem to be not concerned by positive health attributes related to fish consumption. These results may have important implications on production decisions, sales and marketing for the promotion of fish in Lima as a means of economic development.

  2. Implementation of the Fluorescent in Situ Hybridization technique in the Faculty of Medicine, UdelaR

    Directory of Open Access Journals (Sweden)

    Andrea Cairus

    2017-11-01

    Full Text Available The Cytogenetic Laboratory of the Faculty of Medicine processes, on average, 300 annual samples of public and private healthcare centers by conventional cytogenetics. It is essential to implement new techniques to improve the quality of the service offered. The purpose of this work was to implement the Fluorescent in situ Hybridization technique (FISH. An observational, cross-sectional, analytical study was performed. Peripheral blood samples from patients with sex chromosomopathies diagnosed by conventional cytogenetics were analyzed. Fluorescent in situ hybridization technique was applied, comparing results with FISH and with conventional cytogenetics. The percentage of mosaicism detected by conventional cytogenetics and Fluorescent in situ Hybridization was studied: 24 samples were analyzed; 19 presented numerical alterations, 3 structural and 2 both. Numerical alterations were Turner syndrome, Klinefelter syndrome, XXX syndrome and XYY syndrome. Concordance in diagnoses was found for both techniques. For Turner syndrome, 8 of 12 samples corresponded to mosaicism, and there were no significant differences between conventional cytogenetics and the technique studied (p0.05. Klinefelter syndrome and XYY were both presented in a non-mosaic karyotype. For XXX syndrome, a normal line (46, XX was observed in three of the samples, in a percentage close to the cut off. From this research, it will be possible to implement Fluorescent in situ Hybridization in this service, to extend it to other pathologies and to enable the training of human resources; consolidating this laboratory as a national academic reference center.

  3. Performance and lipid profiles of native chickens fed diet containing skipjack fish oil as by-product of fish canning factory

    Science.gov (United States)

    Leke, J. R.; Mandey, J. S.; Laihad, J. T.; Tinangon, R. M.; Tangkau, L.; Junus, C.

    2018-01-01

    The study was conducted to determine the use of fish oil as by-product of fish canning factory in diet on the performance and lipid profiles of native chickens. The experiment used 100 native chicken with an average initial body weight of 48,9 gram (sd + 9.9), was used in this study for 8 weeks experiment. These were arranged by a completely randomized design with 5 treatments, 5 replications and 4 hens in replication each. The diets were: R0 = 100% Based Diet (BD) + 0% Fish Oil (FO); R1 = 98.5% BD + 1.5% FO; R2 = 98% BD + 2% FO; R3 = 97.5% BD + 2.5% FO; R4 = 97 % BD + 3% FO. Feed and water were provided ad libitum. Variables were performance parameters and lipid profiles. Results showed that fish oil inclusion in diets were significantly increased feed intake, body weight gain, carcass percentage, liver, breast and thigh weight, and decreased blood cholesterol, carbohydrate and meat cholesterol, and also tended to decrease abdominal fat. However, there were no affected on feed conversion, water, protein, fat and ash of breast meat. It can be concluded that the use of fish oil in diet up to 3% could improved performance parameters of native chickens.

  4. Problems and solutions in high-rate multichannel hybrid photodiode design The CMS experience

    CERN Document Server

    Cushman, P B

    2002-01-01

    The unique conditions of the CMS experiment (4 T magnetic field, restricted access, high neutron radiation, and 25-ns bunch-crossings) necessitated the development of a new type of high-rate multichannel hybrid photodiode for the tile/fiber hadronic calorimeter. New complexities arose in the push toward high-rate operation, necessitating design changes in the diode structure and surface treatment. The product is now capable of high-rate operation with low crosstalk and leakage current. Lifetime studies of high-voltage behavior, total charge, and irradiation have shown that the tubes will survive the ten years of CMS running with only a few percent change in gain and manageable leakage current rise. (13 refs).

  5. Enlargement of Wallago Leeri with Fish Meal Combination

    OpenAIRE

    Rianti, Evi; Tang, Usman M; Putra, Iskandar

    2014-01-01

    This research was conducted from February to April 2014, for 60 days inexperiment pondof Fisheries Faculty and Marine Sciences, University ofRiauPekanbaru.The aims of the research was todetermine the effect of fish mealcombinations on the growth and survival of Wallago leerireared in ponds.TheMethod used was experiment with3 treatments and 3 replications. The treatmentswereP1= 25% fish trash + 75% pellet, P2 = 50% fish trash + 50% pellet, P3 = 75%fishtrash + 25% pellet.The best result was tre...

  6. Utilization of irradiated sludge for fish feed

    International Nuclear Information System (INIS)

    Harsojo; Syamsu, Suwirma; Subagyo, Lydia Andini

    1994-01-01

    An experiment was conducted to study the use of irradiated sludge pellet for fish feed, namely pellet A consisting of irradiated sludge and shrimp waste (1:3); pellet B consisting of irradiated sludge and commercial pellet (1:2). Pellet C, which is a commercial fish feed, was used as control. Catfish (Clarias gariepinus) was used in this experiment. The feed pellet with a dose of 5% of total body weight was given 3 times per day. The results of the experiments showed that based on food conversion for the relative growth of the catfishes, and heavy metal content, pellet A was the best. No contamination of Salmonella or Shigella bacteria was detected in each pellet. (author). 8 refs, 3 tabs, 1 fig

  7. Gene Expression Profiling in Fish Toxicology: A Review.

    Science.gov (United States)

    Kumar, Girish; Denslow, Nancy D

    In this review, we present an overview of transcriptomic responses to chemical exposures in a variety of fish species. We have discussed the use of several molecular approaches such as northern blotting, differential display reverse transcription-polymerase chain reaction (DDRT-PCR), suppression subtractive hybridization (SSH), real time quantitative PCR (RT-qPCR), microarrays, and next-generation sequencing (NGS) for measuring gene expression. These techniques have been mainly used to measure the toxic effects of single compounds or simple mixtures in laboratory conditions. In addition, only few studies have been conducted to examine the biological significance of differentially expressed gene sets following chemical exposure. Therefore, future studies should focus more under field conditions using a multidisciplinary approach (genomics, proteomics and metabolomics) to understand the synergetic effects of multiple environmental stressors and to determine the functional significance of differentially expressed genes. Nevertheless, recent developments in NGS technologies and decreasing costs of sequencing holds the promise to uncover the complexity of anthropogenic impacts and biological effects in wild fish populations.

  8. Fishing down the largest coral reef fish species.

    Science.gov (United States)

    Fenner, Douglas

    2014-07-15

    Studies on remote, uninhabited, near-pristine reefs have revealed surprisingly large populations of large reef fish. Locations such as the northwestern Hawaiian Islands, northern Marianas Islands, Line Islands, U.S. remote Pacific Islands, Cocos-Keeling Atoll and Chagos archipelago have much higher reef fish biomass than islands and reefs near people. Much of the high biomass of most remote reef fish communities lies in the largest species, such as sharks, bumphead parrots, giant trevally, and humphead wrasse. Some, such as sharks and giant trevally, are apex predators, but others such as bumphead parrots and humphead wrasse, are not. At many locations, decreases in large reef fish species have been attributed to fishing. Fishing is well known to remove the largest fish first, and a quantitative measure of vulnerability to fishing indicates that large reef fish species are much more vulnerable to fishing than small fish. The removal of large reef fish by fishing parallels the extinction of terrestrial megafauna by early humans. However large reef fish have great value for various ecological roles and for reef tourism. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. RNA FISH for detecting expanded repeats in human diseases.

    Science.gov (United States)

    Urbanek, Martyna O; Krzyzosiak, Wlodzimierz J

    2016-04-01

    RNA fluorescence in situ hybridization (FISH) is a widely used technique for detecting transcripts in fixed cells and tissues. Many variants of RNA FISH have been proposed to increase signal strength, resolution and target specificity. The current variants of this technique facilitate the detection of the subcellular localization of transcripts at a single molecule level. Among the applications of RNA FISH are studies on nuclear RNA foci in diseases resulting from the expansion of tri-, tetra-, penta- and hexanucleotide repeats present in different single genes. The partial or complete retention of mutant transcripts forming RNA aggregates within the nucleoplasm has been shown in multiple cellular disease models and in the tissues of patients affected with these atypical mutations. Relevant diseases include, among others, myotonic dystrophy type 1 (DM1) with CUG repeats, Huntington's disease (HD) and spinocerebellar ataxia type 3 (SCA3) with CAG repeats, fragile X-associated tremor/ataxia syndrome (FXTAS) with CGG repeats, myotonic dystrophy type 2 (DM2) with CCUG repeats, amyotrophic lateral sclerosis/frontotemporal dementia (ALS/FTD) with GGGGCC repeats and spinocerebellar ataxia type 32 (SCA32) with GGCCUG. In this article, we summarize the results obtained with FISH to examine RNA nuclear inclusions. We provide a detailed protocol for detecting RNAs containing expanded CAG and CUG repeats in different cellular models, including fibroblasts, lymphoblasts, induced pluripotent stem cells and murine and human neuronal progenitors. We also present the results of the first single-molecule FISH application in a cellular model of polyglutamine disease. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Characterization of genome in tetraploid StY species of Elymus (Triticeae: Poaceae) using sequential FISH and GISH.

    Science.gov (United States)

    Liu, Ruijuan; Wang, Richard R-C; Yu, Feng; Lu, Xingwang; Dou, Quanwen

    2017-08-01

    Genomes of ten species of Elymus, either presumed or known as tetraploid StY, were characterized using fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH). These tetraploid species could be grouped into three categories. Type I included StY genome reported species-Roegneria pendulina, R. nutans, R. glaberrima, R. ciliaris, and Elymus nevskii, and StY genome presumed species-R. sinica, R. breviglumis, and R. dura, whose genome could be separated into two sets based on different GISH intensities. Type I genome constitution was deemed as putative StY. The St genome were mainly characterized with intense hybridization with pAs1, fewer AAG sites, and linked distribution of 5S rDNA and 18S-26S rDNA, while the Y genome with less intense hybridization with pAs1, more varied AAG sites, and isolated distribution of 5S rDNA and 18S-26S rDNA. Nevertheless, further genomic variations were detected among the different StY species. Type II included E. alashanicus, whose genome could be easily separated based on GISH pattern. FISH and GISH patterns suggested that E. alashanicus comprised a modified St genome and an unknown genome. Type III included E. longearistatus, whose genome could not be separated by GISH and was designated as St l Y l . Notably, a close relationship between S l and Y l genomes was observed.

  11. Design of Eco Friendly Shallow Draft Fishing Vessel

    Directory of Open Access Journals (Sweden)

    Sunardi Sunardi

    2016-04-01

    Full Text Available One of the main problem of inland waterways fisheries is the transportation of fish from ponds to fish market during low tide trough inland waterways with 0.6m water depth.The boat is experiences grounding due to water depth of the river is not sufficient for the fishing boat to carry fish at it’s maximum2 tones capacity or experience dead freight . This condition forces fisherman to wait until the high tide from the sea, this delay causes the quality of the fish is decreasing.Besides the problem dead freight  problem the existing vessel is causes environmental problem such as erosion of the river bank due to wake wash. The other important issue is the increases of fuel price and it’s scarcity.  This paper presents the results of comparison of existing monohull fishing boat and two other alternativecatamaran designs. The catamaran design alternatives are is ordinary catamaran and flat side catamaran.  Both of the catamaran fishing boat design shows that the catamaran boat with 0.5m draft is able to carry more than 2 tonnes payload during low tide water depth.  The CFD simulation results shows that flat side catamaran resistance is more than 17.7% lower compared to ordinary catamaran and 44% lower compared to monohull. It means that the consumption of flat side catamaran is lowest compared to two other type of hull design. The flat side catamaran also produces lowest wake wash compared to o two other design. The low wake wash means more friendly to environment.

  12. Waste composting and proving fish for production the organic fertilizers

    Directory of Open Access Journals (Sweden)

    Fernanda San Martins Sanes

    2015-06-01

    Full Text Available The volumes of waste generated in the fishing activity are increasing due to the increase in demand for these products. This implies the need for fast processing and cycling of these materials. Thus, the aim of this study was to evaluate the use of waste generated in the fishing activity as a source of organic fertilizers in agricultural production systems familiar ecological basis. The experiment was conducted at the Experimental Station Cascade / Embrapa Temperate Climate was assessed throughout the composting process and the fermentation of fish waste, identifying the main points that enable the use of these fertilizers in farming systems ecological base. The composting process of rice husk revealed be incomplete during the experiment. The compound prepared with fish waste and exhausted bark of acacia presents itself as a good source of nutrients for crops, which may be suitable as organic fertilizer for production of ecologically-based systems. For liquid organic fertilizer, the conditions under which the experiment was conducted, it is concluded that the compound resulting from aerobic or anaerobic fermentation of fish waste, present themselves as a viable source of nutrients for productive systems of ecological base. However, further studies need to be conducted to better understanding and qualification of both processes.

  13. Research of a hybrid undulator

    International Nuclear Information System (INIS)

    Ma Youwu; Wu Bing; Liu Bo

    1995-12-01

    A 1.5 m tapered hybrid undulator has been designed and built for mid-infrared free electron laser experiments at CIAE. The undulator utilizes the REC-steel hybrid configuration. The magnetic gap and magnetic field taper can be continuously adjusted. The rms error of the peak field is less than 0.53%. The electron trajectory deviation is around 0.03 mm. The design of undulator, sorting of magnets in hybrid undulator using simulated annealing technique, the motion of electron beam in the ideal and measured magnetic field, magnetic field measurement technique and magnetic field adjustment are described. (6 refs., 10 figs., 1 tab)

  14. Detection of ROS1 Gene Rearrangement in Lung Adenocarcinoma: Comparison of IHC, FISH and Real-Time RT-PCR

    OpenAIRE

    Shan, Ling; Lian, Fang; Guo, Lei; Qiu, Tian; Ling, Yun; Ying, Jianming; Lin, Dongmei

    2015-01-01

    Aims To compare fluorescence in situ hybridization (FISH), immunohistochemistry (IHC) and quantitative real-time reverse transcription-PCR (qRT-PCR) assays for detection of ROS1 fusion in a large number of ROS1-positive lung adenocatcinoma (ADC) patients. Methods Using IHC analysis, sixty lung ADCs including 16 cases with ROS1 protein expression and 44 cases without ROS1 expression were selected for this study. The ROS1 fusion status was examined by FISH and qRT-PCR assay. Results Among 60 ca...

  15. Identifying Fishes through DNA Barcodes and Microarrays.

    Directory of Open Access Journals (Sweden)

    Marc Kochzius

    2010-09-01

    Full Text Available International fish trade reached an import value of 62.8 billion Euro in 2006, of which 44.6% are covered by the European Union. Species identification is a key problem throughout the life cycle of fishes: from eggs and larvae to adults in fisheries research and control, as well as processed fish products in consumer protection.This study aims to evaluate the applicability of the three mitochondrial genes 16S rRNA (16S, cytochrome b (cyt b, and cytochrome oxidase subunit I (COI for the identification of 50 European marine fish species by combining techniques of "DNA barcoding" and microarrays. In a DNA barcoding approach, neighbour Joining (NJ phylogenetic trees of 369 16S, 212 cyt b, and 447 COI sequences indicated that cyt b and COI are suitable for unambiguous identification, whereas 16S failed to discriminate closely related flatfish and gurnard species. In course of probe design for DNA microarray development, each of the markers yielded a high number of potentially species-specific probes in silico, although many of them were rejected based on microarray hybridisation experiments. None of the markers provided probes to discriminate the sibling flatfish and gurnard species. However, since 16S-probes were less negatively influenced by the "position of label" effect and showed the lowest rejection rate and the highest mean signal intensity, 16S is more suitable for DNA microarray probe design than cty b and COI. The large portion of rejected COI-probes after hybridisation experiments (>90% renders the DNA barcoding marker as rather unsuitable for this high-throughput technology.Based on these data, a DNA microarray containing 64 functional oligonucleotide probes for the identification of 30 out of the 50 fish species investigated was developed. It represents the next step towards an automated and easy-to-handle method to identify fish, ichthyoplankton, and fish products.

  16. Radiation disinfestation studies on sun dried fish

    International Nuclear Information System (INIS)

    Ahmed, M.; Bhuiya, A.D.; Alam, M.S.; Huda, S.M.S.

    1989-01-01

    A large quantity of dried fish is lost in Bangladesh due to infestation by earwigs, hide beetles and copra beetles in storage godowns. The most destructive pest is the hide beetle, Dermestes maculatus Deg. Earwigs of different developmental stages were exposed to 0.10, 0.20 or 0.30 kGy of gamma rays for disinfestation. There was apparent damage to the control but all treated samples were in good condition and no live insect was observed 3 weeks after starting the experiment. Disinfestation studies of dried mackerel showed that eggs, larval and pupal stages of hide beetles could be inactivated at a dose of 0.20 kGy. A dose of 0.30 kGy killed all adults 2 weeks after irradiation. In the packaging studies, dried mackerel was packed in polythene pouches of different thicknesses. Two controls were maintained, i.e. dried fish with no treatment, control, and dried fish disinfested with heat at 60 deg. C, disinfested control. In experiments with 50 μm thick polythene pouches, the dried fish of irradiation treatments with 0.10 to 1.0 kGy doses and the controls had around 20% moisture content with the exception of disinfested controls which had 13% moisture content. All irradiated samples were free from insect damage. There was heavy damage in the controls due to insects. However, all these treated fish had heavy fungal growth with the exception of the disinfested control. Similar results were obtained with pouches made of 75 μm thick polythene irradiated at doses of 0.50, 1.0, 2.0 and 4.0 kGy. In the final experiments pouches were made of 50, 75 and 100 μm thick polythene and exposed to similar radiation as in the previous experiment. In all the treatments, moisture content was reduced to 13%. heavy insect damage was observed in the control, while all the treated samples were in excellent condition after 5 months of storage. (author). 20 refs, 2 figs, 5 tabs

  17. Laboratory Studies on the Effects of Shear on Fish

    Energy Technology Data Exchange (ETDEWEB)

    Neitzel, Duane A.; Richmond, Marshall C.; Dauble, Dennis D.; Mueller, Robert P.; Moursund, Russell A.; Abernethy, Cary S.; Guensch, Greg R.

    2000-09-20

    The overall objective of our studies was to specify an index describing the hydraulic force that fish experience when subjected to a shear environment. Fluid shear is a phenomenon that is important to fish. However, elevated levels of shear may result in strain rates that injure or kill fish. At hydroelectric generating facilities, concerns have been expressed that strain rates associated with passage through turbines, spillways, and fish bypass systems may adversely affect migrating fish. Development of fish friendly hydroelectric turbines requires knowledge of the physical forces (injury mechanisms) that impact entrained fish and the fish's tolerance to these forces. It requires up-front, pre-design specifications for the environmental conditions that occur within the turbine system, in other words, determining or assuming that those conditions known to injure fish will provide the descriptions of conditions that engineers must consider in the design of a turbine system. These biological specifications must be carefully and thoroughly documented throughout the design of a fish friendly turbine. To address the development of biological specifications, we designed and built a test facility where juvenile fish could be subjected to a range of shear environments and quantified their biological response.

  18. Utilization of geothermal energy for drying fish products

    International Nuclear Information System (INIS)

    Arason, S.; Arnason, H.

    1992-01-01

    This paper is about industrial uses of geothermal energy for drying of fish products. Drying is an ancient method for preservation of foods, the main purpose of which is to increase the preservation time. For drying, an external source of energy is needed to extract water. In this paper an emphasis is placed on drying fish and associated processes, and how geothermal energy can be used to substitute oil or electricity. The Icelandic Fisheries Laboratories have been experimenting with different methods of drying, and several drying stations have been designed for indoor drying of fish products. Today there are more than a dozen companies in this country which are drying fish indoors using for that purpose electricity and/or geothermal energy. Further possibilities are available when fish processing plants are located in geothermal areas

  19. Body temperature change characteristics of Lake Michigan fishes

    International Nuclear Information System (INIS)

    Spigarelli, S.A.; Thommes, M.M.; Beitinger, T.L.

    1974-01-01

    Body temperature change rate experiments were conducted on alewife, brown trout, rainbow trout, brook trout, and carp collected from the discharge flumes and inshore areas near the Point Beach Nuclear Plant. Test fish were exposed to immediate water temperature changes of up to 10.6 0 C by transfer between ambient and discharge water holding tanks. Results showed that the temperature change rate was related to fish size, species, and direction of change, suggesting that rapid temperature changes would have a more pronounced effect on smaller fish

  20. Towards a holistic assessment of the user experience with hybrid BCIs.

    Science.gov (United States)

    Lorenz, Romy; Pascual, Javier; Blankertz, Benjamin; Vidaurre, Carmen

    2014-06-01

    In recent years, brain-computer interfaces (BCIs) have become mature enough to immensely benefit from the expertise and tools established in the field of human-computer interaction (HCI). One of the core objectives in HCI research is the design of systems that provide a pleasurable user experience (UX). While the majority of BCI studies exclusively evaluate common efficiency measures such as classification accuracy and speed, single research groups have begun to look at further usability aspects such as ease of use, workload and learnability. However, these evaluation metrics only cover pragmatic aspects of UX while still not considering the hedonic quality of UX. In order to gain a holistic perspective on UX, hedonic quality aspects such as motivation and frustration were also taken into account for our evaluation of three BCI-driven interfaces, which were proposed to be used as a two-stage neuroprosthetic control within the EU project MUNDUS. At the first stage, one of six possible actions was selected and either confirmed or cancelled at the second stage. For the experiment, a solely event-related-potential-based interface (ERP-ERP) and two hybrid solutions were tested that were controlled by ERP and motor imagery (MI)--resulting in the two possible combinations: ERP selection/MI confirmation (ERP-MI) or MI selection/ERP confirmation (MI-ERP). Behavioural, subjective and encephalographic (EEG) data of 12 healthy subjects were collected during an online experiment with the three graphical user interfaces (GUIs). Results showed a significantly greater pragmatic quality (in terms of accuracy, efficiency, workload, use quality and learnability) for the ERP-ERP and ERP-MI GUIs in contrast to the MI-ERP GUI. Consequently, the MI-ERP GUI is least suited for use as a neuroprosthetic control. With respect to the comparison of the ERP-ERP and ERP-MI GUIs, no significant differences in pragmatic and hedonic quality of UX were found. Since throughout better results were

  1. Towards a holistic assessment of the user experience with hybrid BCIs

    Science.gov (United States)

    Lorenz, Romy; Pascual, Javier; Blankertz, Benjamin; Vidaurre, Carmen

    2014-06-01

    Objective. In recent years, brain-computer interfaces (BCIs) have become mature enough to immensely benefit from the expertise and tools established in the field of human-computer interaction (HCI). One of the core objectives in HCI research is the design of systems that provide a pleasurable user experience (UX). While the majority of BCI studies exclusively evaluate common efficiency measures such as classification accuracy and speed, single research groups have begun to look at further usability aspects such as ease of use, workload and learnability. However, these evaluation metrics only cover pragmatic aspects of UX while still not considering the hedonic quality of UX. In order to gain a holistic perspective on UX, hedonic quality aspects such as motivation and frustration were also taken into account for our evaluation of three BCI-driven interfaces, which were proposed to be used as a two-stage neuroprosthetic control within the EU project MUNDUS. Approach. At the first stage, one of six possible actions was selected and either confirmed or cancelled at the second stage. For the experiment, a solely event-related-potential-based interface (ERP-ERP) and two hybrid solutions were tested that were controlled by ERP and motor imagery (MI)—resulting in the two possible combinations: ERP selection/MI confirmation (ERP-MI) or MI selection/ERP confirmation (MI-ERP). Behavioural, subjective and encephalographic (EEG) data of 12 healthy subjects were collected during an online experiment with the three graphical user interfaces (GUIs). Main results. Results showed a significantly greater pragmatic quality (in terms of accuracy, efficiency, workload, use quality and learnability) for the ERP-ERP and ERP-MI GUIs in contrast to the MI-ERP GUI. Consequently, the MI-ERP GUI is least suited for use as a neuroprosthetic control. With respect to the comparison of the ERP-ERP and ERP-MI GUIs, no significant differences in pragmatic and hedonic quality of UX were found

  2. Flying fish accelerate at 5 G to leap from the water surface

    Science.gov (United States)

    Yang, Patricia; Phonekeo, Sulisay; Xu, Ke; Chang, Shui-Kai; Hu, David

    2013-11-01

    Flying fish can both swim underwater and glide in air. Transitioning from swimming to gliding requires penetration of the air-water interface, or breaking the ``surface tension barrier,'' a formidable task for juvenile flying fish measuring 1 to 5 cm in length. In this experimental investigation, we use high-speed videography to characterize the kinematics of juvenile flying fish as they leap from the water surface. During this process, which lasts 0.05 seconds, flying fish achieve body accelerations of 5 times earth's gravity and gliding speeds of 1.3 m/s, an order of magnitude higher than their steady swimming speed. We rationalize this anomalously high speed on the basis of the hydrodynamic and surface tension forces and torques experienced by the fish. Specifically, leaping fish experience skin friction forces only on the submerged part of their body, permitting them to achieve much higher speeds than in steady underwater swimming. We also perform experiments using a towed flying fish mimc to determine optimality of various parameters in this process, including body angle and start position with respect to the water surface.

  3. Evaluation of HER2 gene amplification in invasive breast cancer using a dual-color chromogenic in situ hybridization (dual CISH).

    Science.gov (United States)

    Kato, Nobuaki; Itoh, Hitoshi; Serizawa, Akihiko; Hatanaka, Yutaka; Umemura, Shinobu; Osamura, R Yoshiyuki

    2010-07-01

    Fluorescence in situ hybridization (FISH) assay is considered the 'gold standard' for evaluation of HER2/neu (HER2) gene status, however, it is difficult to recognize morphologic features of tumors using fluorescence microscopy. Thus, chromogenic in situ hybridization (CISH) has been proposed as an alternative method to evaluate HER2 gene amplification. Here, we examined the dual color CISH (dual CISH) method which provides information regarding the copy number of the HER2 gene and chromosome 17 centromere from a single slide. We examined 40 cases of invasive ductal carcinomas of the breast that were resected surgically. HER2 gene status was assessed with FISH (Abbott) and dual CISH (Dako). HER2 gene amplification status was classified according to the guidelines of the American Society of Clinical Oncology and College of American Pathologists (ASCO/CAP). Comparison of the cut-off values for HER2/chromosome 17 centromere copy number ratio obtained by dual CISH and FISH showed that there was almost perfect agreement between two methods (Kappa coefficient 0.96). The results of the two commercial products were almost consistent for evaluation of HER2 gene counts on the sections. The current study proved that dual CISH is comparable with FISH for evaluating HER2 gene status.

  4. Hybrid stage I palliation for hypo-plastic left heart condition without a hybrid suite: Suggestions for developing nations

    Directory of Open Access Journals (Sweden)

    S. Anuradha

    2012-07-01

    Full Text Available Cardiac hybrid procedures are performed in modern, spacious, and highly equipped hybrid suites in developed countries. Organizing such expensive suites in countries with an emerging economy is difficult from both a financial and logistics point of view. We share our experience of safely performing a Hybrid stage I palliation procedure for Aortic atresia with ventricular septal defect on a 2-month-old infant weighing 3.35 kg using minimal resources in a conventional catheterization laboratory.

  5. Changes in reproductive biomarkers in an endangered fish species (bonytail chub, Gila elegans) exposed to low levels of organic wastewater compounds in a controlled experiment.

    Science.gov (United States)

    Walker, David B; Paretti, Nicholas V; Cordy, Gail; Gross, Timothy S; Zaugg, Steven D; Furlong, Edward T; Kolpin, Dana W; Matter, William J; Gwinn, Jessica; McIntosh, Dennis

    2009-11-08

    In arid regions of the southwestern United States, municipal wastewater treatment plants commonly discharge treated effluent directly into streams that would otherwise be dry most of the year. A better understanding is needed of how effluent-dependent waters (EDWs) differ from more natural aquatic ecosystems and the ecological effect of low levels of environmentally persistent organic wastewater compounds (OWCs) with distance from the pollutant source. In a controlled experiment, we found 26 compounds common to municipal effluent in treatment raceways all at concentrations fish. Female bonytail chub in treatment tanks had significantly lower concentrations of 17beta-estradiol than control females (p=0.001). The normally inverse relationship between primary male and female sex hormones, expected in un-impaired fish, was greatly decreased in treatment (r=0.00) versus control (r=-0.66) female fish. We found a similar, but not as significant, trend between treatment (r=-0.45) and control (r=-0.82) male fish. Measures of fish condition showed no significant differences between male or female fish housed in effluent or clean water. Inter-sex condition did not occur and testicular and ovarian cells appeared normal for the respective developmental stage and we observed no morphological alteration in fish. The population-level impacts of these findings are uncertain. Studies examining the long-term, generational and behavioral effects to aquatic organisms chronically exposed to low levels of OWC mixtures are needed.

  6. Imaging of Chromosome Dynamics in Mouse Testis Tissue by Immuno-FISH.

    Science.gov (United States)

    Scherthan, Harry

    2017-01-01

    The mouse (Mus musculus) represents the central mammalian genetic model system for biomedical and developmental research. Mutant mouse models have provided important insights into chromosome dynamics during the complex meiotic differentiation program that compensates for the genome doubling at fertilization. Homologous chromosomes (homologues) undergo dynamic pairing and recombine during first meiotic prophase before they become partitioned into four haploid sets by two consecutive meiotic divisions that lack an intervening S-phase. Fluorescence in situ hybridization (FISH) has been instrumental in the visualization and imaging of the dynamic reshaping of chromosome territories and mobility during prophase I, in which meiotic telomeres were found to act as pacemakers for the chromosome pairing dance. FISH combined with immunofluorescence (IF) co-staining of nuclear proteins has been instrumental for the visualization and imaging of mammalian meiotic chromosome behavior. This chapter describes FISH and IF methods for the analysis of chromosome dynamics in nuclei of paraffin-embedded mouse testes. The techniques have proven useful for fresh and archived paraffin testis material of several mammalian species.

  7. Passive appendages improve the maneuverability of fish-like robots

    Science.gov (United States)

    Pollard, Beau; Tallapragada, Phanindra

    2017-11-01

    It is known that the passive mechanics of fish appendages play a role in the high efficiency of their swimming. A well known example of this is the experimental demonstration that a dead fish could swim upstream. However little is known about the role if any of passive deformations of a fish-like body that could aid in its maneuverability. Part of the difficulty investigating this lies in clearly separating the role of actuated body deformations and passive deformations in response to the fluid structure interaction. In this paper we compare the maneuverability of several fish shaped robotic models that possess varying numbers of passive appendages with a fish shaped robot that has no appendages. All the robots are propelled by the oscillations of an internal momentum wheel thereby eliminating any active deformations of the body. Our experiments clearly reveal the significant improvement in maneuverability of robots with passive appendages. In the broader context of swimming robots our experiments show that passive mechanisms could be useful to provide mechanical feedback that can help maneuverability and obstacle avoidance along with propulsive efficiency. This work was partly supported by a Grant from the NSF CMMI 1563315.

  8. Detection of MDM2/CDK4 amplification in lipomatous soft tissue tumors from formalin-fixed, paraffin-embedded tissue: comparison of multiplex ligation-dependent probe amplification (MLPA) and fluorescence in situ hybridization (FISH).

    Science.gov (United States)

    Creytens, David; van Gorp, Joost; Ferdinande, Liesbeth; Speel, Ernst-Jan; Libbrecht, Louis

    2015-02-01

    In this study, the detection of MDM2 and CDK4 amplification was evaluated in lipomatous soft tissue tumors using multiplex ligation-dependent probe amplification (MLPA), a PCR-based technique, in comparison with fluorescence in situ hybridization (FISH). These 2 techniques were evaluated in a series of 77 formalin-fixed, paraffin-embedded lipomatous tumors (27 benign adipose tumors, 28 atypical lipomatous tumors/well-differentiated liposarcomas, 18 dedifferentiated liposarcomas, and 4 pleomorphic liposarcomas). Using MLPA, with a cut-off ratio of >2, 36/71 samples (22 atypical lipomatous tumors/well-differentiated liposarcomas, and 14 dedifferentiated liposarcomas) showed MDM2 and CDK4 amplification. Using FISH as gold standard, MLPA showed a sensitivity of 90% (36/40) and a specificity of 100% (31/31) in detecting amplification of MDM2 and CDK4 in lipomatous soft tissue tumors. In case of high-level amplification (MDM2-CDK4/CEP12 ratio >5), concordance was 100%. Four cases of atypical lipomatous tumor/well-differentiated liposarcoma (4/26, 15%) with a low MDM2 and CDK4 amplification level (MDM2-CDK4/CEP12 ratio ranging between 2 and 2.5) detected by FISH showed no amplification by MLPA, although gain of MDM2 and CDK4 (ratios ranging between 1.6 and 1.9) was seen with MLPA. No amplification was detected in benign lipomatous tumors and pleomorphic liposarcomas. Furthermore, there was a very high concordance between the ratios obtained by FISH and MLPA. In conclusion, MLPA proves to be an appropriate and straightforward technique for screening MDM2/CDK4 amplification in lipomatous tumors, especially when a correct cut-off value and reference samples are chosen, and could be considered a good alternative to FISH to determine MDM2 and CDK4 amplification in liposarcomas. Moreover, because MLPA, as a multiplex technique, allows simultaneous detection of multiple chromosomal changes of interest, it could be in the future a very reliable and fast molecular analysis on

  9. The virtual lover: variable and easily guided 3D fish animations as an innovative tool in mate-choice experiments with sailfin mollies-II. Validation.

    Science.gov (United States)

    Gierszewski, Stefanie; Müller, Klaus; Smielik, Ievgen; Hütwohl, Jan-Marco; Kuhnert, Klaus-Dieter; Witte, Klaudia

    2017-02-01

    The use of computer animation in behavioral research is a state-of-the-art method for designing and presenting animated animals to live test animals. The major advantages of computer animations are: (1) the creation of animated animal stimuli with high variability of morphology and even behavior; (2) animated stimuli provide highly standardized, controlled and repeatable testing procedures; and (3) they allow a reduction in the number of live test animals regarding the 3Rs principle. But the use of animated animals should be attended by a thorough validation for each test species to verify that behavior measured with live animals toward virtual animals can also be expected with natural stimuli. Here we present results on the validation of a custom-made simulation for animated 3D sailfin mollies Poecilia latipinna and show that responses of live test females were as strong to an animated fish as to a video or a live male fish. Movement of an animated stimulus was important but female response was stronger toward a swimming 3D fish stimulus than to a "swimming" box. Moreover, male test fish were able to discriminate between animated male and female stimuli; hence, rendering the animated 3D fish a useful tool in mate-choice experiments with sailfin mollies.

  10. Hybrid Design, Procurement and Testing for the LHCb Silicon Tracker

    CERN Document Server

    Bay, A; Frei, R; Jiménez-Otero, S; Perrin, A; Tran, MT; Van Hunen, J J; Vervink, K; Vollhardt, A; Agari, M; Bauer, C; Blouw, J; Hofmann, W; Knöpfle, K T; Löchner, S; Schmelling, M; Schwingenheuer, B; Smale, N J; Adeva, B; Esperante-Pereira, D; Lois, C; Vázquez, P; Lehner, F; Bernhard, R P; Bernet, R; Gassner, J; Köstner, S; Needham, M; Steinkamp, O; Straumann, U; Volyanskyy, D; Voss, H; Wenger, A

    2005-01-01

    The Silicon Tracker of the LHCb experiment consists of four silicon detector stations positioned along the beam line of the experiment. The detector modules of each station are constructed from wide pitch silicon microstrip sensors. Located at the module's end, a polyimide hybrid is housing the front-end electronics. The assembly of the more than 600 hybrids has been outsourced to industry. We will report on the design and production status of the hybrids for the LHCb Silicon Tracker and describe the quality assurance tests. Particular emphasis is laid on the vendor qualifying and its impact on our hybrid design that we experienced during the prototyping phase.

  11. Accuracy Assessment of Interphase Fluorescence In-Situ Hybridization on Uncultured Amniotic Fluid Cells

    Directory of Open Access Journals (Sweden)

    Hamideh Karimi

    2007-01-01

    Full Text Available Background: Parental anxiety while waiting for the results of amniocentesis has been investigatedby many authors. It seems that the implementation of faster techniques such as fluorescence in-situhybridization (FISH will have some benefits in reducing this anxiety. Besides the patients' attitudesto choosing this method, gynecologists who are the persons responsible for treatment, must feelcomfortable about prescribing FISH techniques.Materials and Methods: This study, using a simple methodology, was undertaken to evaluate theresults of FISH tests on the amniotic fluid from 40 pregnant women undergoing cesarean surgery.Two sets of probes including X/Y cocktail and 13, 21 and 18 were applied on different slides.Results: The results of FISH tests were compared with the reports of the pediatrician about thehealth condition of the newborn. Complete conformity between the two sets of findings, haveconvinced our gynecologists of the benefit of prescribing this method to reduce the anxiety ofpatients at risk of having abnormal offspring due to chromosomal anuploidies.Conclusion: As has been documented by many authors, conventional chromosome analysis hasgreat advantages over fluorescence in situ hybridization of interphase amniocytes, but reducing theanxiety of parents is a good reason for employing the FISH technique.

  12. X chromosome control of meiotic chromosome synapsis in mouse inter-subspecific hybrids.

    Directory of Open Access Journals (Sweden)

    Tanmoy Bhattacharyya

    2014-02-01

    Full Text Available Hybrid sterility (HS belongs to reproductive isolation barriers that safeguard the integrity of species in statu nascendi. Although hybrid sterility occurs almost universally among animal and plant species, most of our current knowledge comes from the classical genetic studies on Drosophila interspecific crosses or introgressions. With the house mouse subspecies Mus m. musculus and Mus m. domesticus as a model, new research tools have become available for studies of the molecular mechanisms and genetic networks underlying HS. Here we used QTL analysis and intersubspecific chromosome substitution strains to identify a 4.7 Mb critical region on Chromosome X (Chr X harboring the Hstx2 HS locus, which causes asymmetrical spermatogenic arrest in reciprocal intersubspecific F1 hybrids. Subsequently, we mapped autosomal loci on Chrs 3, 9 and 13 that can abolish this asymmetry. Combination of immunofluorescent visualization of the proteins of synaptonemal complexes with whole-chromosome DNA FISH on pachytene spreads revealed that heterosubspecific, unlike consubspecific, homologous chromosomes are predisposed to asynapsis in F1 hybrid male and female meiosis. The asynapsis is under the trans- control of Hstx2 and Hst1/Prdm9 hybrid sterility genes in pachynemas of male but not female hybrids. The finding concurred with the fertility of intersubpecific F1 hybrid females homozygous for the Hstx2(Mmm allele and resolved the apparent conflict with the dominance theory of Haldane's rule. We propose that meiotic asynapsis in intersubspecific hybrids is a consequence of cis-acting mismatch between homologous chromosomes modulated by the trans-acting Hstx2 and Prdm9 hybrid male sterility genes.

  13. X chromosome control of meiotic chromosome synapsis in mouse inter-subspecific hybrids.

    Science.gov (United States)

    Bhattacharyya, Tanmoy; Reifova, Radka; Gregorova, Sona; Simecek, Petr; Gergelits, Vaclav; Mistrik, Martin; Martincova, Iva; Pialek, Jaroslav; Forejt, Jiri

    2014-02-01

    Hybrid sterility (HS) belongs to reproductive isolation barriers that safeguard the integrity of species in statu nascendi. Although hybrid sterility occurs almost universally among animal and plant species, most of our current knowledge comes from the classical genetic studies on Drosophila interspecific crosses or introgressions. With the house mouse subspecies Mus m. musculus and Mus m. domesticus as a model, new research tools have become available for studies of the molecular mechanisms and genetic networks underlying HS. Here we used QTL analysis and intersubspecific chromosome substitution strains to identify a 4.7 Mb critical region on Chromosome X (Chr X) harboring the Hstx2 HS locus, which causes asymmetrical spermatogenic arrest in reciprocal intersubspecific F1 hybrids. Subsequently, we mapped autosomal loci on Chrs 3, 9 and 13 that can abolish this asymmetry. Combination of immunofluorescent visualization of the proteins of synaptonemal complexes with whole-chromosome DNA FISH on pachytene spreads revealed that heterosubspecific, unlike consubspecific, homologous chromosomes are predisposed to asynapsis in F1 hybrid male and female meiosis. The asynapsis is under the trans- control of Hstx2 and Hst1/Prdm9 hybrid sterility genes in pachynemas of male but not female hybrids. The finding concurred with the fertility of intersubpecific F1 hybrid females homozygous for the Hstx2(Mmm) allele and resolved the apparent conflict with the dominance theory of Haldane's rule. We propose that meiotic asynapsis in intersubspecific hybrids is a consequence of cis-acting mismatch between homologous chromosomes modulated by the trans-acting Hstx2 and Prdm9 hybrid male sterility genes.

  14. X Chromosome Control of Meiotic Chromosome Synapsis in Mouse Inter-Subspecific Hybrids

    Science.gov (United States)

    Bhattacharyya, Tanmoy; Reifova, Radka; Gregorova, Sona; Simecek, Petr; Gergelits, Vaclav; Mistrik, Martin; Martincova, Iva; Pialek, Jaroslav; Forejt, Jiri

    2014-01-01

    Hybrid sterility (HS) belongs to reproductive isolation barriers that safeguard the integrity of species in statu nascendi. Although hybrid sterility occurs almost universally among animal and plant species, most of our current knowledge comes from the classical genetic studies on Drosophila interspecific crosses or introgressions. With the house mouse subspecies Mus m. musculus and Mus m. domesticus as a model, new research tools have become available for studies of the molecular mechanisms and genetic networks underlying HS. Here we used QTL analysis and intersubspecific chromosome substitution strains to identify a 4.7 Mb critical region on Chromosome X (Chr X) harboring the Hstx2 HS locus, which causes asymmetrical spermatogenic arrest in reciprocal intersubspecific F1 hybrids. Subsequently, we mapped autosomal loci on Chrs 3, 9 and 13 that can abolish this asymmetry. Combination of immunofluorescent visualization of the proteins of synaptonemal complexes with whole-chromosome DNA FISH on pachytene spreads revealed that heterosubspecific, unlike consubspecific, homologous chromosomes are predisposed to asynapsis in F1 hybrid male and female meiosis. The asynapsis is under the trans- control of Hstx2 and Hst1/Prdm9 hybrid sterility genes in pachynemas of male but not female hybrids. The finding concurred with the fertility of intersubpecific F1 hybrid females homozygous for the Hstx2Mmm allele and resolved the apparent conflict with the dominance theory of Haldane's rule. We propose that meiotic asynapsis in intersubspecific hybrids is a consequence of cis-acting mismatch between homologous chromosomes modulated by the trans-acting Hstx2 and Prdm9 hybrid male sterility genes. PMID:24516397

  15. Testing the 'hybrid susceptibility' and 'phenological sink' hypotheses using the P. balsamifera - P. deltoides hybrid zone and septoria leaf spot [Septoria musiva].

    Science.gov (United States)

    LeBoldus, Jared M; Isabel, Nathalie; Floate, Kevin D; Blenis, Peter; Thomas, Barb R

    2013-01-01

    Hybrid genotypes that arise between plant species frequently have increased susceptibility to arthropod pests and fungal pathogens. This pattern has been attributed to the breakdown of plant defenses ('Hybrid susceptibility' hypothesis) and (or) to extended periods of susceptibility attributed to plant phenologies in zones of species overlap and (or) hybridization ('phenological sink' hypothesis). We examined these hypotheses by assessing the susceptibility of parental and hybrid Populus host genotypes to a leaf spot disease caused by the fungal pathogen Septoria musiva. For this purpose, 214 genotypes were obtained from morphologically pure zones of P. balsamifera and P. deltoides, and from an intervening zone of overlap and hybridization on the drainage of the Red Deer River, Alberta, Canada. Genotypes were identified as P. balsamifera, P. deltoides, or hybrid using a suite of 27 species-specific SNP markers. Initially the genetic structure of the hybrid zone was characterized with 27.7% of trees classified as admixed individuals. To test the hybrid susceptibility hypothesis, a subset of 52 genotypes was inoculated with four isolates of S. musiva. Levels of susceptibility were P. balsamifera > F1 hybrid > P. deltoides. A further 53 genotypes were grown in a common garden to assess the effect of genotype on variation in leaf phenology. Leaf phenology was more variable within the category of hybrid genotypes than within categories of either parental species. Leaf phenology was also more variable for the category of trees originating in the hybrid (P. balsamifera - P. deltoides [hybrid and parental genotypes combined]) zone than in adjacent pure zones of the parental species. The results from the inoculation experiment support the hybrid intermediacy hypothesis. The results from the common garden experiment support the 'phenological sink' hypothesis. These findings have greatly increased our understanding of the epidemiology and ecology of fungal pathogens in plant

  16. Testing the 'hybrid susceptibility' and 'phenological sink' hypotheses using the P. balsamifera - P. deltoides hybrid zone and septoria leaf spot [Septoria musiva].

    Directory of Open Access Journals (Sweden)

    Jared M LeBoldus

    Full Text Available Hybrid genotypes that arise between plant species frequently have increased susceptibility to arthropod pests and fungal pathogens. This pattern has been attributed to the breakdown of plant defenses ('Hybrid susceptibility' hypothesis and (or to extended periods of susceptibility attributed to plant phenologies in zones of species overlap and (or hybridization ('phenological sink' hypothesis. We examined these hypotheses by assessing the susceptibility of parental and hybrid Populus host genotypes to a leaf spot disease caused by the fungal pathogen Septoria musiva. For this purpose, 214 genotypes were obtained from morphologically pure zones of P. balsamifera and P. deltoides, and from an intervening zone of overlap and hybridization on the drainage of the Red Deer River, Alberta, Canada. Genotypes were identified as P. balsamifera, P. deltoides, or hybrid using a suite of 27 species-specific SNP markers. Initially the genetic structure of the hybrid zone was characterized with 27.7% of trees classified as admixed individuals. To test the hybrid susceptibility hypothesis, a subset of 52 genotypes was inoculated with four isolates of S. musiva. Levels of susceptibility were P. balsamifera > F1 hybrid > P. deltoides. A further 53 genotypes were grown in a common garden to assess the effect of genotype on variation in leaf phenology. Leaf phenology was more variable within the category of hybrid genotypes than within categories of either parental species. Leaf phenology was also more variable for the category of trees originating in the hybrid (P. balsamifera - P. deltoides [hybrid and parental genotypes combined] zone than in adjacent pure zones of the parental species. The results from the inoculation experiment support the hybrid intermediacy hypothesis. The results from the common garden experiment support the 'phenological sink' hypothesis. These findings have greatly increased our understanding of the epidemiology and ecology of fungal

  17. FISH DETECTION OF CAMPYLOBACTER AND ARCOBACTER ADHERED TO STAINLESS STEEL COUPONS

    Directory of Open Access Journals (Sweden)

    Lucie Šilhová

    2015-02-01

    Full Text Available This study focuses on detecting biofilm and planktonic bacterial cells of the genera Arcobacter and Campylobacter. This study is, to our knowledge, the first study deals with application of FISH procedure to detect biofilm formation on stainless steel coupons of Arcobacter-isolates from real-world environments. These bacteria can cause a lot of diseases. Especially, in the last decade, arcobacters have been increasingly isolated from feces of clinically healthy and ill animals, foods of animal origin and various types of water. Fluorescence in situ hybridization was used to detect biofilm and planktonic cells of selected microorganisms. This method was optimized and subsequently applied to biofilm samples prepared on stainless steel coupons. The study results indicate that fluorescence in situ hybridization is suitable for detecting biofilm and planktonic cells of the studied bacteria.

  18. Salinity and temperature tolerance of an emergent alien species, the Amazon fish Astronotus ocellatus

    Science.gov (United States)

    Gutierrel, Silvia M M; Schofield, Pam; Prodocimo, Viviane

    2016-01-01

    Astronotus ocellatus (oscar), is native to the Amazon basin and, although it has been introduced to many countries, little is known regarding its tolerances for salinity and temperature. In this report, we provide data on the tolerance of A. ocellatus to abrupt and gradual changes in salinity, its high and low temperature tolerance, and information on how salinity, temperature, and fish size interact to affect survival. Fish were able to survive abrupt transfer to salinities as high as 16 ppt with no mortality. When salinity change was gradual (2 ppt/day), fish in the warm-temperature experiment (28°C) survived longer than fish in the cool-temperature experiment (18°C). Larger fish survived longer than smaller ones at the higher salinities when the temperature was warm, but when the temperature was cool fish size had little effect on survival. In the temperature-tolerance experiments, fish survived from 9 to 41°C for short periods of time. Overall, the species showed a wide range of temperature and salinity tolerance. Thus, in spite of the tropical freshwater origin of this species, physiological stress is not likely to hinder its dispersal to brackish waters, especially when temperatures are warm.

  19. Dual-colour CISH is a reliable alternative to FISH for assessment of topoisomerase 2-alpha amplification in breast carcinomas.

    Science.gov (United States)

    García-Caballero, Tomás; Prieto, Olga; Vázquez-Boquete, Angel; Gude, Francisco; Viaño, Patricia; Otero, María; Curiel, Teresa; Fernández-Rodríguez, Beatriz; Parrado, Concepción; Fraga, Máximo; Antúnez, José R

    2014-01-01

    Anthracyclines are among the most powerful antineoplastic drugs available for breast cancer treatment. Although HER2 amplification has been postulated to predict anthracycline benefit, numerous reports have demonstrated that HER2/TOP2A co-amplification is the clinically useful predictive marker of response to anthracyclines. The standard technique to evaluate gene status for target therapy selection is fluorescence in situ hybridization (FISH), but this technique has some disadvantages. Dual-colour chromogenic in situ hybridization (CISH) is an extension of the FISH protocol that allows bright-field microscopy and thus represents a user-friendly alternative to FISH. In order to evaluate whether dual-colour CISH is a reliable alternative to FISH in determining TOP2A gene amplification and to determine the frequency with which TOP2A and HER2 were co-amplified, we analysed 100 invasive breast cancer specimens (70 consecutive and 30 HER2-amplified samples) using tissue microarrays. Thus, a 99 % agreement was found between TOP2A status determined by dual-colour CISH and FISH, as well as a high degree of correlation in TOP2A ratios using both techniques. TOP2A gene amplification was present in 8.6 % of the 70 consecutive samples studied, all of which were HER2-amplified. Co-amplification of TOP2A was observed in 46.5 % of the additional 30 HER2-amplified samples (no TOP2A amplification was seen in non-amplified HER2 samples). We conclude that dual-colour CISH represents an excellent alternative to FISH for determination of TOP2A gene status in invasive breast cancer. Our results showing TOP2A amplification only in HER2-amplified cases also add to the evidence that TOP2A determination should be restricted to those cases.

  20. Improving Health Care Coverage, Equity, And Financial Protection Through A Hybrid System: Malaysia's Experience.

    Science.gov (United States)

    Rannan-Eliya, Ravindra P; Anuranga, Chamara; Manual, Adilius; Sararaks, Sondi; Jailani, Anis S; Hamid, Abdul J; Razif, Izzanie M; Tan, Ee H; Darzi, Ara

    2016-05-01

    Malaysia has made substantial progress in providing access to health care for its citizens and has been more successful than many other countries that are better known as models of universal health coverage. Malaysia's health care coverage and outcomes are now approaching levels achieved by member nations of the Organization for Economic Cooperation and Development. Malaysia's results are achieved through a mix of public services (funded by general revenues) and parallel private services (predominantly financed by out-of-pocket spending). We examined the distributional aspects of health financing and delivery and assessed financial protection in Malaysia's hybrid system. We found that this system has been effective for many decades in equalizing health care use and providing protection from financial risk, despite modest government spending. Our results also indicate that a high out-of-pocket share of total financing is not a consistent proxy for financial protection; greater attention is needed to the absolute level of out-of-pocket spending. Malaysia's hybrid health system presents continuing unresolved policy challenges, but the country's experience nonetheless provides lessons for other emerging economies that want to expand access to health care despite limited fiscal resources. Project HOPE—The People-to-People Health Foundation, Inc.

  1. Detection of reciprocal chromosome translocations as an indicator of organism exposure to ionizing radiation by FISH-WCP method

    International Nuclear Information System (INIS)

    Holeckova, B.; Sivikova, K.; Dianovsky, J.; Piesova, E.; Lakatosova, M.

    2006-01-01

    Chromosome translocations are considered to be the gold standard for assessing ionizing radiation exposure. Because translocations are inherently more stable through cell division than dicentrics, translocations have become the aberration of choice for evaluating many types of exposure. Fluorescence in situ hybridization with whole chromosome painting probes (FISH-WCP) has been shown to be a rapid method of detecting chromosomal rearrangements, and appears to be especially useful for analysis of induced translocations. The present paper shortly describes FISH-WCP method for detection of reciprocal translocations as indicators of exposure to ionizing radiation. (authors)

  2. Fish-allergic patients may be able to eat fish.

    Science.gov (United States)

    Mourad, Ahmad A; Bahna, Sami L

    2015-03-01

    Reported fish allergy prevalence varies widely, with an estimated prevalence of 0.2% in the general population. Sensitization to fish can occur by ingestion, skin contact or inhalation. The manifestations can be IgE or non-IgE mediated. Several fish allergens have been identified, with parvalbumins being the major allergen in various species. Allergenicity varies among fish species and is affected by processing or preparation methods. Adverse reactions after eating fish are often claimed to be 'allergy' but could be a reaction to hidden food allergen, fish parasite, fish toxins or histamine in spoiled fish. Identifying such causes would allow free consumption of fish. Correct diagnosis of fish allergy, including the specific species, might provide the patient with safe alternatives. Patients have been generally advised for strict universal avoidance of fish. However, testing with various fish species or preparations might identify one or more forms that can be tolerated.

  3. Feasibility of iFISH patterns in hematologic malignancies among Congolese patients at Kinshasa University clinics

    Directory of Open Access Journals (Sweden)

    Mireille Solange Nganga Nkanga

    2017-12-01

    Full Text Available Objective: To analyze the feasibility of detecting Ph1 in leukemia patients in the Kinshasa University Clinics in the Democratic Republic of Congo, at KU Leuven, Belgium. Methods: Bone marrow and peripheral blood samples with chronic myeloid leukemia, acute myeloid leukemia or acute leukocytes leukemia were obtained from 32 patients in Kinshasa University clinics in the Democratic Republic of Congo and transferred to KU Leuven in Belgium for iFISH feasibility. Ph1 was detected by using a remote analysis of interphase fluorescence in situ hybridization (iFISH. Results: Out of the 32 patients involved in this study, 65.6% (n = 21 of the cases were successfully tested, of which 52.4% (n = 11 were iFISH positives for the variant t(9;22 (presence of Ph1 in chronic myeloid leukemia samples and 47.6% (n = 10 negatives in all subtypes of hematological malignancies. However, there was a female predominance in chronic myeloid leukemia samples Ph1-positives by iFISH, whereas no sexual influence was observed on acute subtypes of leukemia. Conclusions: iFISH analysis is feasible on samples obtained from remote sites in the Democratic Republic of Congo. However, the optimization of the sample storage is necessary to further improve iFISH's performance. Keywords: iFISH, Ph1, Democratic Republic of Congo, Leukemia, Bone marrow, Blood

  4. Can environmental impact assessments alone conserve freshwater fish biota? Review of the Chilean experience

    International Nuclear Information System (INIS)

    Lacy, Shaw Nozaki; Meza, Francisco J.; Marquet, Pablo A.

    2017-01-01

    Chile was one of many countries that initiated environmental impact assessments in the 1990s, and has relied on their use for species conservation and territorial planning without the use of larger-scale environmental and ecological planning. The capacity of Chile's environmental impact assessment system (SEIA) to evaluate resident freshwater fishes and the potential impacts of water projects and aquaculture activities – two categories of projects that create direct threats to freshwater fishes – are assessed. Of the 3997 such submissions to the SEIA, only 0.6% conducted any freshwater fish assessment, and only 0.1% conducted any quantitative assessment of expected impacts from the associated project. The small number of assessments was characterized by poor study design, inconsistent sampling methodology, and species misidentification. Traditional assessments failed to include freshwater fish ecology in the general assessment framework. The new strategic environmental evaluation system only underscores the need for vastly improved field sampling protocols and assessment methodologies.

  5. Can environmental impact assessments alone conserve freshwater fish biota? Review of the Chilean experience

    Energy Technology Data Exchange (ETDEWEB)

    Lacy, Shaw Nozaki, E-mail: shaw.lacy@gmail.com [Departamento de Ecología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Alameda 340, Santiago (Chile); Departmento de Ecosistemas y Medio Ambiente, Pontificia Universidad Católica de Chile, Vicuña Mackenna 4860, Macul (Chile); Centro Interdisciplinario de Cambio Global, Vicuña Mackenna 4860, Macul (Chile); Meza, Francisco J. [Departmento de Ecosistemas y Medio Ambiente, Pontificia Universidad Católica de Chile, Vicuña Mackenna 4860, Macul (Chile); Centro Interdisciplinario de Cambio Global, Vicuña Mackenna 4860, Macul (Chile); Marquet, Pablo A. [Departamento de Ecología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Alameda 340, Santiago (Chile); Instituto de Ecología y Biodiversidad, Las Palmeras 345, Santiago (Chile); Centro Interdisciplinario de Cambio Global, Vicuña Mackenna 4860, Macul (Chile)

    2017-03-15

    Chile was one of many countries that initiated environmental impact assessments in the 1990s, and has relied on their use for species conservation and territorial planning without the use of larger-scale environmental and ecological planning. The capacity of Chile's environmental impact assessment system (SEIA) to evaluate resident freshwater fishes and the potential impacts of water projects and aquaculture activities – two categories of projects that create direct threats to freshwater fishes – are assessed. Of the 3997 such submissions to the SEIA, only 0.6% conducted any freshwater fish assessment, and only 0.1% conducted any quantitative assessment of expected impacts from the associated project. The small number of assessments was characterized by poor study design, inconsistent sampling methodology, and species misidentification. Traditional assessments failed to include freshwater fish ecology in the general assessment framework. The new strategic environmental evaluation system only underscores the need for vastly improved field sampling protocols and assessment methodologies.

  6. Mapping of single-copy genes by TSA-FISH in the codling moth, Cydia pomonella.

    Science.gov (United States)

    Carabajal Paladino, Leonela Z; Nguyen, Petr; Síchová, Jindra; Marec, František

    2014-01-01

    We work on the development of transgenic sexing strains in the codling moth, Cydia pomonella (Tortricidae), which would enable to produce male-only progeny for the population control of this pest using sterile insect technique (SIT). To facilitate this research, we have developed a number of cytogenetic and molecular tools, including a physical map of the codling moth Z chromosome using BAC-FISH (fluorescence in situ hybridization with bacterial artificial chromosome probes). However, chromosomal localization of unique, single-copy sequences such as a transgene cassette by conventional FISH remains challenging. In this study, we adapted a FISH protocol with tyramide signal amplification (TSA-FISH) for detection of single-copy genes in Lepidoptera. We tested the protocol with probes prepared from partial sequences of Z-linked genes in the codling moth. Using a modified TSA-FISH protocol we successfully mapped a partial sequence of the Acetylcholinesterase 1 (Ace-1) gene to the Z chromosome and confirmed thus its Z-linkage. A subsequent combination of BAC-FISH with BAC probes containing anticipated neighbouring Z-linked genes and TSA-FISH with the Ace-1 probe allowed the integration of Ace-1 in the physical map of the codling moth Z chromosome. We also developed a two-colour TSA-FISH protocol which enabled us simultaneous localization of two Z-linked genes, Ace-1 and Notch, to the expected regions of the Z chromosome. We showed that TSA-FISH represents a reliable technique for physical mapping of genes on chromosomes of moths and butterflies. Our results suggest that this technique can be combined with BAC-FISH and in the future used for physical localization of transgene cassettes on chromosomes of transgenic lines in the codling moth or other lepidopteran species. Furthermore, the developed protocol for two-colour TSA-FISH might become a powerful tool for synteny mapping in non-model organisms.

  7. Fish Allergy

    Science.gov (United States)

    ... Cause Blog Vision Awards Common Allergens Fish Allergy Fish Allergy Learn about fish allergy, how to read ... that you must avoid both. Allergic Reactions to Fish Finned fish can cause severe and potentially life- ...

  8. Fluorescence in situ hybridization as adjunct to cytology improves the diagnosis and directs estimation of prognosis of malignant pleural effusions

    Directory of Open Access Journals (Sweden)

    Han Jingquan

    2012-11-01

    Full Text Available Abstract Background The identification of malignant cells in effusions by conventional cytology is hampered by its limited sensitivity and specificity. The aim of this study was to investigate the value of fluorescence in situ hybridization (FISH as adjuncts to conventional cytologic examination in patients with malignant pleural effusions. Methods We conducted a retrospective cohort study of 93 inpatients with pleural effusions (72 malignant pleural effusions metastatic from 11 different organs and 21 benign over 23 months. All the patients came from Chinese northeast areas. Aspirated pleural fluid underwent cytologic examination and fluorescence in situ hybridization (FISH for aneuploidy. We used FISH in single-colour or if appropriate in dual-colour evaluation to detect chromosomal aberrations (chromosomes 7, 11, and 17 in effusion cells as markers of malignancy, to raise the diagnostic yield and identified the efficiency by diagnostic biopsy. Predominant cytogenetic anomalies and patterns of intratumor cytogenetic heterogeneity were brought in relation to overall survival rate. Results Cytology alone confirmed malignant pleural effusions in 45 of 72 patients (sensitivity 63%, whereas FISH alone positively identified 48 of 72 patients (sensitivity 67%. Both tests had high specificity in predicting benign effusions. If cytology and FISH were considered together, they exhibited 88% sensitivity and 94.5% specificity in discriminating benign and malignant effusions. Combined, the two assays were more sensitive than either test alone. Although the positive predictive value of each test was 94.5%, the negative predictive value of cytology and FISH combined was 78%, better than 47% and 44% for FISH and cytology alone, respectively. There was a significantly prolonged survival rate for patients with aneuploidy for chromosome 17. Conclusions FISH in combination with conventional cytology is a highly sensitive and specific diagnostic tool for detecting

  9. A Hybrid Approach to Teaching Managerial Economics

    Science.gov (United States)

    Metzgar, Matthew

    2014-01-01

    Many institutions in higher education are experimenting with hybrid teaching approaches to undergraduate courses. Online resources may provide a number of advantages to students as compared to in-class approaches. Research regarding the effectiveness of hybrid approaches is mixed and still accumulating. This paper discusses the use of a hybrid…

  10. Frequency of reciprocal translocations and dicentrics induced in human blood lymphocytes by X-irradiation as determined by fluorescence in situ hybridization

    International Nuclear Information System (INIS)

    Nakano, M.; Nakashima, E.; Pawel, D.; Kodama, Y.; Awa, A.

    1993-01-01

    This study was designed to test the scoring efficiency of reciprocal translocations and dicentrics induced by X-irradiation in vitro using the fluorescence in situ hybridization (FISH) technique. An excess was found in the frequencies of reciprocal translocations relative to those of dicentrics by measurement with FISH at the first cell division after irradiation (translocation:dicentric ≅ 60:40). However, when the same metaphases were also evaluated sequentially by a conventional staining method, the ratio of about 50:50 was restored. This was due in part to misclassification of certain dicentrics as reciprocal translocations by the FISH technique. (author)

  11. Managing conflicts arising from fisheries enhancements based on non-native fishes in southern Africa.

    Science.gov (United States)

    Ellender, B R; Woodford, D J; Weyl, O L F; Cowx, I G

    2014-12-01

    Southern Africa has a long history of non-native fish introductions for the enhancement of recreational and commercial fisheries, due to a perceived lack of suitable native species. This has resulted in some important inland fisheries being based on non-native fishes. Regionally, these introductions are predominantly not benign, and non-native fishes are considered one of the main threats to aquatic biodiversity because they affect native biota through predation, competition, habitat alteration, disease transfer and hybridization. To achieve national policy objectives of economic development, food security and poverty eradication, countries are increasingly looking towards inland fisheries as vehicles for development. As a result, conflicts have developed between economic and conservation objectives. In South Africa, as is the case for other invasive biota, the control and management of non-native fishes is included in the National Environmental Management: Biodiversity Act. Implementation measures include import and movement controls and, more recently, non-native fish eradication in conservation priority areas. Management actions are, however, complicated because many non-native fishes are important components in recreational and subsistence fisheries that contribute towards regional economies and food security. In other southern African countries, little attention has focussed on issues and management of non-native fishes, and this is cause for concern. This paper provides an overview of introductions, impacts and fisheries in southern Africa with emphasis on existing and evolving legislation, conflicts, implementation strategies and the sometimes innovative approaches that have been used to prioritize conservation areas and manage non-native fishes. © 2014 The Fisheries Society of the British Isles.

  12. Six-degree-of-freedom Sensor Fish design - Governing equations and motion modeling

    Energy Technology Data Exchange (ETDEWEB)

    Zeng, D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Richmond, M. C. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Simmons, C. S. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Carlson, T. J. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2004-07-01

    The Sensor Fish device is being used at Northwest hydropower projects to better understand the conditions fish experience during passage through hydro turbines and other dam bypass alternatives. Since its initial development in 1997, the Sensor Fish has undergone numerous design changes to improve its function and extend the range of its use. The most recent Sensor Fish design, the three degree of freedom (3DOF) device, has been used successfully to characterize the environment fish experience when passing through turbines, in spill, or in engineered fish bypass facilities at dams. Pacific Northwest National Laboratory (PNNL) is in the process of redesigning the current 3DOF Sensor Fish device package to improve its field performance. Rate gyros will be added to the new six degree of freedom (6DOF) device so that it will be possible to observe the six linear and angular accelerations of the Sensor Fish as it passes the dam. Before the 6DOF Sensor Fish device can be developed and deployed, governing equations of motion must be developed in order to understand the design implications of instrument selection and placement within the body of the device. The report describes a fairly general formulation for the coordinate systems, equations of motion, force and moment relationships necessary to simulate the 6DOF movement of an underwater body.

  13. Identification and determination of the viability of Giardia lamblia cysts and Cryptosporidium parvum and Cryptosporidium hominis oocysts in human fecal and water supply samples by fluorescent in situ hybridization (FISH) and monoclonal antibodies.

    Science.gov (United States)

    Lemos, Vanessa; Graczyk, Thaddeus K; Alves, Margarida; Lobo, Maria Luísa; Sousa, Maria C; Antunes, Francisco; Matos, Olga

    2005-12-01

    In the present study, fluorescent in situ hybridization (FISH) and monoclonal antibodies (MAbs) were evaluated for species-specific detection and viability determination of Giardia lamblia, Cryptosporidium parvum, and Cryptosporidium hominis in human fecal and water supply samples. A total of 50 fecal human samples positive for G. lamblia cysts, 38 positive for C. parvum, and 23 positive for C. hominis were studied. Also, 18 water supply samples positive for Giardia spp. and Cryptosporidium spp. by the United States Environmental Protection Agency (USEPA) Method 1623 were studied by FISH and fluorescein isothiocyanate (FITC)-conjugated MAbs. Eighteen percent of the fecal samples parasitologically positive for G. lamblia presented viable and nonviable cysts, and 5% of those positive for Cryptosporidium spp. presented viable and nonviable oocysts. Of the 18 water supply samples analyzed, 6 (33%) presented Giardia spp. viable and nonviable cysts and 2 (11%) presented viable and nonviable Cryptosporidium spp. oocysts. G. lamblia identification was confirmed by polymerase chain reaction (PCR) and sequencing of the beta-giardin gene in the fecal and water samples found positive by FISH and FITC-conjugated MAbs. C. parvum and Cryptosporidium muris were identified, by PCR and sequencing of the small subunit of ribosomal RNA gene, in seven and one water samples, respectively. Our results confirm that this technique enables simultaneous visualization, species-specific identification, and viability determination of the organisms present in human fecal and water supply samples.

  14. Radioactivity in the pelagic fish. I. Distribution of radioactivity in various tissues of fish

    Energy Technology Data Exchange (ETDEWEB)

    Amano, K; Yamada, K; Bito, M; Takase, A; Tanaka, S

    1955-01-01

    Pelagic fishes caught after an atomic explosion experiment at Bikini Atolls in the Pacific were examined by radiochemical techniques. Generally the radioactivity was large in liver, kidney, gall bladder and heart, and then in pyloric ceca, stomach, intestine, and gonad; there was little activity in skin, bone, and muscles. This order varied with species. Large radioactivity of the stomach contents did not necessarily mean large activity in the tissues, indicating considerable participation of diffusion of sea water into the fish body. Muscles from various sites showed slight difference in the activity. The dark muscle, however, showed several times as large activity as ordinary muscle.

  15. Low-contrast underwater living fish recognition using PCANet

    Science.gov (United States)

    Sun, Xin; Yang, Jianping; Wang, Changgang; Dong, Junyu; Wang, Xinhua

    2018-04-01

    Quantitative and statistical analysis of ocean creatures is critical to ecological and environmental studies. And living fish recognition is one of the most essential requirements for fishery industry. However, light attenuation and scattering phenomenon are present in the underwater environment, which makes underwater images low-contrast and blurry. This paper tries to design a robust framework for accurate fish recognition. The framework introduces a two stage PCA Network to extract abstract features from fish images. On a real-world fish recognition dataset, we use a linear SVM classifier and set penalty coefficients to conquer data unbalanced issue. Feature visualization results show that our method can avoid the feature distortion in boundary regions of underwater image. Experiments results show that the PCA Network can extract discriminate features and achieve promising recognition accuracy. The framework improves the recognition accuracy of underwater living fishes and can be easily applied to marine fishery industry.

  16. Trophic relationships of small nonnative fishes in a natural creek and several agricultural drains flowing into the Salton Sea, and their potential, effects on the endangered desert pupfish

    Science.gov (United States)

    Martin, Barbara A.; Saiki, Michael K.

    2009-01-01

    This study was conducted to characterize trophic relationships of small nonnative fishes and to determine if predation by these fishes contributes to the decline of desert pupfish (Cyprinodon macularius), an endangered cyprinodont on the verge of extinction. We sampled 403 hybrid Mozambique tilapias (Oreochromis mossambica by O. urolepis), 107 redbelly tilapias (Tilapia zillii), 32 longjaw mudsuckers (Gillkhthys mirabilis), 182 western mosquitofish (Gambusia affinis), 222 sailfin mollies (Poecilia latipinna), 63 shortfin mollies (Poecilia mexicana), and 235 porthole livebearers (Poecilurpsis gracilis) from a natural creek and four agricultural drains during September 1999- December 2001. Evidence of piscivory was in gastrointestinal contents of 14 hybrid Mozambique tilapias, 3 redbelly tilapias, 10 longjaw mudsuckers, 8 western mosquitofish, 2 sailfin mollies, and 8 porthole livebearers. Although digestion often was too advanced for identification of fishes consumed by nonnative fishes, remains of desert pupfish were in gastrointestinal contents of a longjaw mudsucker. Our findings, along with Field evidence from other studies that inverse relationships exist between abundances of desert pupfish and nonnative species, are consistent with the hypothesis that predation by nonnative species is contributing to decline of desert pupfish. We suspect that competitive interactions with nonnative fishes might also adversely affect abundance of desert pupfish.

  17. 49 CFR 173.218 - Fish meal or fish scrap.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 2 2010-10-01 2010-10-01 false Fish meal or fish scrap. 173.218 Section 173.218... Fish meal or fish scrap. (a) Except as provided in Column (7) of the HMT in § 172.101 of this subchapter, fish meal or fish scrap, containing at least 6%, but not more than 12% water, is authorized for...

  18. PV-hybrid and mini-grid

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2010-07-01

    ) Optimization of a wind/diesel hybrid configuration in a remote grid with battery implementation: Case study of Melinka Island; (23) Provisional acceptance of installations and online data submission of PV and hybrid kits in remote areas of Latin-America under the EC's EURO-SOLAR programme; (24) Experience of the Canary Islands in the development of insular 100 % RES systems and micro-grids; (25) Assessment of photovoltaic hybrid power systems in the United States; (26) Solar hybrid school project in East Malaysia; (27) Eigg Island - Electrification of a British Island by a unique PV wind hydro diesel hybrid system; (28) A pragmatic performance reporting approach for describing PV hybrid systems within mini-grids: Work in progress from IEA's PVPS Task 11 Act. 31; (29) Hybrid renewable energy systems for the supply of services in rural settlements of Mediterranean partner countries. The HYRESS project - The case study of the hybrid system - Micro grid in Egypt. Beside these lectures, the following poster contributions were presented: (1) Performance of conventional MPPT techniques in the presence of partial shielding; (2) Photovoltaic and thermal collector (PV/T) hybrid system's performance analysis under the mild climate conditions of Izmir City; (3) Influential parameters on a building integrated hybrid PVT concentrator; (4) The solution to combine and manage renewable energies in hybrid applications and mini-grids; (5) Stabilization of distribution networks with PV and vanadium redox-battery backup systems - Simulation and first experiences; (6) Control, monitoring and data acquisition architecture design for clean production of hydrogen from mini-wind energy; (7) Remote Telecom System including photovoltaic energy and H{sub 2} production by electrolysis; (8) Effective combination of solar and wind energy systems; (9) Standardisation of distributed grid support - An analogous approach for the smart grid; (10) Optimizing energy management of decentralized

  19. Judging a salmon by its spots: environmental variation is the primary determinant of spot patterns in Salmo salar.

    Science.gov (United States)

    Jørgensen, Katarina M; Solberg, Monica F; Besnier, Francois; Thorsen, Anders; Fjelldal, Per Gunnar; Skaala, Øystein; Malde, Ketil; Glover, Kevin A

    2018-04-12

    In fish, morphological colour changes occur from variations in pigment concentrations and in the morphology, density, and distribution of chromatophores in the skin. However, the underlying mechanisms remain unresolved in most species. Here, we describe the first investigation into the genetic and environmental basis of spot pattern development in one of the world's most studied fishes, the Atlantic salmon. We reared 920 salmon from 64 families of domesticated, F1-hybrid and wild origin in two contrasting environments (Hatchery; tanks for the freshwater stage and sea cages for the marine stage, and River; a natural river for the freshwater stage and tanks for the marine stage). Fish were measured, photographed and spot patterns evaluated. In the Hatchery experiment, significant but modest differences in spot density were observed among domesticated, F1-hybrid (1.4-fold spottier than domesticated) and wild salmon (1.7-fold spottier than domesticated). A heritability of 6% was calculated for spot density, and a significant QTL on linkage group SSA014 was detected. In the River experiment, significant but modest differences in spot density were also observed among domesticated, F1-hybrid (1.2-fold spottier than domesticated) and wild salmon (1.8-fold spottier than domesticated). Domesticated salmon were sevenfold spottier in the Hatchery vs. River experiment. While different wild populations were used for the two experiments, on average, these were 6.2-fold spottier in the Hatchery vs. River experiment. Fish in the Hatchery experiment displayed scattered to random spot patterns while fish in the River experiment displayed clustered spot patterns. These data demonstrate that while genetics plays an underlying role, environmental variation represents the primary determinant of spot pattern development in Atlantic salmon.

  20. Environmental effects on fish neural plasticity and cognition.

    Science.gov (United States)

    Ebbesson, L O E; Braithwaite, V A

    2012-12-01

    Most fishes experiencing challenging environments are able to adjust and adapt their physiology and behaviour to help them cope more effectively. Much of this flexibility is supported and influenced by cognition and neural plasticity. The understanding of fish cognition and the role played by different regions of the brain has improved significantly in recent years. Techniques such as lesioning, tract tracing and quantifying changes in gene expression help in mapping specialized brain areas. It is now recognized that the fish brain remains plastic throughout a fish's life and that it continues to be sensitive to environmental challenges. The early development of fish brains is shaped by experiences with the environment and this can promote positive and negative effects on both neural plasticity and cognitive ability. This review focuses on what is known about the interactions between the environment, the telencephalon and cognition. Examples are used from a diverse array of fish species, but there could be a lot to be gained by focusing research on neural plasticity and cognition in fishes for which there is already a wealth of knowledge relating to their physiology, behaviour and natural history, e.g. the Salmonidae. © 2012 The Authors. Journal of Fish Biology © 2012 The Fisheries Society of the British Isles.

  1. Sexual ornaments, body morphology, and swimming performance in naturally hybridizing swordtails (teleostei: xiphophorus.

    Directory of Open Access Journals (Sweden)

    James B Johnson

    Full Text Available Determining the costs of sexual ornaments is complicated by the fact that ornaments are often integrated with other, non-sexual traits, making it difficult to dissect the effect of ornaments independent of other aspects of the phenotype. Hybridization can produce reduced phenotypic integration, allowing one to evaluate performance across a broad range of multivariate trait values. Here we assess the relationship between morphology and performance in the swordtails Xiphophorus malinche and X. birchmanni, two naturally-hybridizing fish species that differ extensively in non-sexual as well as sexual traits. We took advantage of novel trait variation in hybrids to determine if sexual ornaments incur a cost in terms of locomotor ability. For both fast-start and endurance swimming, hybrids performed at least as well as the two parental species. The sexually-dimorphic sword did not impair swimming performance per se. Rather, the sword negatively affected performance only when paired with a sub-optimal body shape. Studies seeking to quantify the costs of ornaments should consider that covariance with non-sexual traits may create the spurious appearance of costs.

  2. Characterization of microbial associations with methanotrophic archaea and sulfate-reducing bacteria through statistical comparison of nested Magneto-FISH enrichments

    Directory of Open Access Journals (Sweden)

    Elizabeth Trembath-Reichert

    2016-04-01

    Full Text Available Methane seep systems along continental margins host diverse and dynamic microbial assemblages, sustained in large part through the microbially mediated process of sulfate-coupled Anaerobic Oxidation of Methane (AOM. This methanotrophic metabolism has been linked to consortia of anaerobic methane-oxidizing archaea (ANME and sulfate-reducing bacteria (SRB. These two groups are the focus of numerous studies; however, less is known about the wide diversity of other seep associated microorganisms. We selected a hierarchical set of FISH probes targeting a range of Deltaproteobacteria diversity. Using the Magneto-FISH enrichment technique, we then magnetically captured CARD-FISH hybridized cells and their physically associated microorganisms from a methane seep sediment incubation. DNA from nested Magneto-FISH experiments was analyzed using Illumina tag 16S rRNA gene sequencing (iTag. Enrichment success and potential bias with iTag was evaluated in the context of full-length 16S rRNA gene clone libraries, CARD-FISH, functional gene clone libraries, and iTag mock communities. We determined commonly used Earth Microbiome Project (EMP iTAG primers introduced bias in some common methane seep microbial taxa that reduced the ability to directly compare OTU relative abundances within a sample, but comparison of relative abundances between samples (in nearly all cases and whole community-based analyses were robust. The iTag dataset was subjected to statistical co-occurrence measures of the most abundant OTUs to determine which taxa in this dataset were most correlated across all samples. Many non-canonical microbial partnerships were statistically significant in our co-occurrence network analysis, most of which were not recovered with conventional clone library sequencing, demonstrating the utility of combining Magneto-FISH and iTag sequencing methods for hypothesis generation of associations within complex microbial communities. Network analysis pointed to

  3. Characterization of microbial associations with methanotrophic archaea and sulfate-reducing bacteria through statistical comparison of nested Magneto-FISH enrichments.

    Science.gov (United States)

    Trembath-Reichert, Elizabeth; Case, David H; Orphan, Victoria J

    2016-01-01

    Methane seep systems along continental margins host diverse and dynamic microbial assemblages, sustained in large part through the microbially mediated process of sulfate-coupled Anaerobic Oxidation of Methane (AOM). This methanotrophic metabolism has been linked to consortia of anaerobic methane-oxidizing archaea (ANME) and sulfate-reducing bacteria (SRB). These two groups are the focus of numerous studies; however, less is known about the wide diversity of other seep associated microorganisms. We selected a hierarchical set of FISH probes targeting a range of Deltaproteobacteria diversity. Using the Magneto-FISH enrichment technique, we then magnetically captured CARD-FISH hybridized cells and their physically associated microorganisms from a methane seep sediment incubation. DNA from nested Magneto-FISH experiments was analyzed using Illumina tag 16S rRNA gene sequencing (iTag). Enrichment success and potential bias with iTag was evaluated in the context of full-length 16S rRNA gene clone libraries, CARD-FISH, functional gene clone libraries, and iTag mock communities. We determined commonly used Earth Microbiome Project (EMP) iTAG primers introduced bias in some common methane seep microbial taxa that reduced the ability to directly compare OTU relative abundances within a sample, but comparison of relative abundances between samples (in nearly all cases) and whole community-based analyses were robust. The iTag dataset was subjected to statistical co-occurrence measures of the most abundant OTUs to determine which taxa in this dataset were most correlated across all samples. Many non-canonical microbial partnerships were statistically significant in our co-occurrence network analysis, most of which were not recovered with conventional clone library sequencing, demonstrating the utility of combining Magneto-FISH and iTag sequencing methods for hypothesis generation of associations within complex microbial communities. Network analysis pointed to many co

  4. Hybridization-Based Detection of Helicobacter pylori at Human Body Temperature Using Advanced Locked Nucleic Acid (LNA) Probes

    Science.gov (United States)

    Fontenete, Sílvia; Guimarães, Nuno; Leite, Marina; Figueiredo, Céu; Wengel, Jesper; Filipe Azevedo, Nuno

    2013-01-01

    The understanding of the human microbiome and its influence upon human life has long been a subject of study. Hence, methods that allow the direct detection and visualization of microorganisms and microbial consortia (e.g. biofilms) within the human body would be invaluable. In here, we assessed the possibility of developing a variant of fluorescence in situ hybridization (FISH), named fluorescence in vivo hybridization (FIVH), for the detection of Helicobacter pylori. Using oligonucleotide variations comprising locked nucleic acids (LNA) and 2’-O-methyl RNAs (2’OMe) with two types of backbone linkages (phosphate or phosphorothioate), we were able to successfully identify two probes that hybridize at 37 °C with high specificity and sensitivity for H. pylori, both in pure cultures and in gastric biopsies. Furthermore, the use of this type of probes implied that toxic compounds typically used in FISH were either found to be unnecessary or could be replaced by a non-toxic substitute. We show here for the first time that the use of advanced LNA probes in FIVH conditions provides an accurate, simple and fast method for H. pylori detection and location, which could be used in the future for potential in vivo applications either for this microorganism or for others. PMID:24278398

  5. Fish Ontology framework for taxonomy-based fish recognition

    Science.gov (United States)

    Ali, Najib M.; Khan, Haris A.; Then, Amy Y-Hui; Ving Ching, Chong; Gaur, Manas

    2017-01-01

    Life science ontologies play an important role in Semantic Web. Given the diversity in fish species and the associated wealth of information, it is imperative to develop an ontology capable of linking and integrating this information in an automated fashion. As such, we introduce the Fish Ontology (FO), an automated classification architecture of existing fish taxa which provides taxonomic information on unknown fish based on metadata restrictions. It is designed to support knowledge discovery, provide semantic annotation of fish and fisheries resources, data integration, and information retrieval. Automated classification for unknown specimens is a unique feature that currently does not appear to exist in other known ontologies. Examples of automated classification for major groups of fish are demonstrated, showing the inferred information by introducing several restrictions at the species or specimen level. The current version of FO has 1,830 classes, includes widely used fisheries terminology, and models major aspects of fish taxonomy, grouping, and character. With more than 30,000 known fish species globally, the FO will be an indispensable tool for fish scientists and other interested users. PMID:28929028

  6. Fish Ontology framework for taxonomy-based fish recognition

    Directory of Open Access Journals (Sweden)

    Najib M. Ali

    2017-09-01

    Full Text Available Life science ontologies play an important role in Semantic Web. Given the diversity in fish species and the associated wealth of information, it is imperative to develop an ontology capable of linking and integrating this information in an automated fashion. As such, we introduce the Fish Ontology (FO, an automated classification architecture of existing fish taxa which provides taxonomic information on unknown fish based on metadata restrictions. It is designed to support knowledge discovery, provide semantic annotation of fish and fisheries resources, data integration, and information retrieval. Automated classification for unknown specimens is a unique feature that currently does not appear to exist in other known ontologies. Examples of automated classification for major groups of fish are demonstrated, showing the inferred information by introducing several restrictions at the species or specimen level. The current version of FO has 1,830 classes, includes widely used fisheries terminology, and models major aspects of fish taxonomy, grouping, and character. With more than 30,000 known fish species globally, the FO will be an indispensable tool for fish scientists and other interested users.

  7. COMBO-FISH Enables High Precision Localization Microscopy as a Prerequisite for Nanostructure Analysis of Genome Loci

    Directory of Open Access Journals (Sweden)

    Rainer Kaufmann

    2010-10-01

    Full Text Available With the completeness of genome databases, it has become possible to develop a novel FISH (Fluorescence in Situ Hybridization technique called COMBO-FISH (COMBinatorial Oligo FISH. In contrast to other FISH techniques, COMBO-FISH makes use of a bioinformatics approach for probe set design. By means of computer genome database searching, several oligonucleotide stretches of typical lengths of 15–30 nucleotides are selected in such a way that all uniquely colocalize at the given genome target. The probes applied here were Peptide Nucleic Acids (PNAs—synthetic DNA analogues with a neutral backbone—which were synthesized under high purity conditions. For a probe repetitively highlighted in centromere 9, PNAs labeled with different dyes were tested, among which Alexa 488Ò showed reversible photobleaching (blinking between dark and bright state a prerequisite for the application of SPDM (Spectral Precision Distance/Position Determination Microscopy a novel technique of high resolution fluorescence localization microscopy. Although COMBO-FISH labeled cell nuclei under SPDM conditions sometimes revealed fluorescent background, the specific locus was clearly discriminated by the signal intensity and the resulting localization accuracy in the range of 10–20 nm for a detected oligonucleotide stretch. The results indicate that COMBO-FISH probes with blinking dyes are well suited for SPDM, which will open new perspectives on molecular nanostructural analysis of the genome.

  8. Development of an observation robot `Flying Fish` for comprehensive measurements of ocean environment; Kaiyo kankyo sogo kansoku robot `flying fish` no kaihatsu kenkyu

    Energy Technology Data Exchange (ETDEWEB)

    Koterayama, W.; Yamaguchi, S.; Nakamura, M. [Kyushu Univ., Fukuoka (Japan). Research Inst. for Applied Mechanics; Akamatsu, T. [Mitsubishi Heavy Industries, Ltd., Tokyo (Japan)

    1996-12-31

    With an objective for spatial continuous measurement of physical, chemical and biological amounts in ocean, development has been made on a wing controlled towed vehicle, `Flying Fish` which is capable of controlling depth, pitch and roll. Numerical simulations and two-year sea experiments have been carried out for the development. Flying Fish consists of a sub-system comprising a towing vehicle, towing cables, and on-board controllers. In a steady state, Flying Fish can be controlled at accuracy for depth of {plus_minus} 0.05m, pitch of {plus_minus} one degree, and roll of {plus_minus} 0.5 degree. This accuracy is sufficient for operating a chemical analyzer, the dissolved carbonic acid analyzer. Even in a non-steady state such as in changing the depth, the pitch can be controlled at {plus_minus} 3 degrees and the roll at {plus_minus} 0.5 degree. This extent of attitude change is within a range rendering no problems in maintaining accuracy of the measurement devices. The result of sea experiments for movements of Flying Fish agreed with that of the numerical simulation at practically usable accuracy. Flying Fish is verified as an effective system in investigating spatial variations in ocean data. 10 refs., 15 figs., 1 tab.

  9. Prognostic classification of MDS is improved by the inclusion of FISH panel testing with conventional cytogenetics.

    Science.gov (United States)

    Kokate, Prajakta; Dalvi, Rupa; Koppaka, Neeraja; Mandava, Swarna

    2017-10-01

    Cytogenetics is a critical independent prognostic factor in myelodysplastic syndromes (MDS). Conventional cytogenetics (CC) and Fluorescence in situ hybridization (FISH) Panel Testing are extensively used for the prognostic stratification of MDS, although the FISH test is not yet a bona fide component of the International Prognostic Scoring System (IPSS). The present study compares the utility of CC and FISH to detect chromosomal anomalies and in prognostic categorization. GTG-Banding and FISH Panel Testing specifically for -5/-5q, -7/-7q, +8 and -20q was performed on whole blood or bone marrow samples from 136 patients with MDS. Chromosomal anomalies were found in 40 cases by CC, including three novel translocations. FISH identified at least one anomaly in 54/136 (39.7%) cases. More than one anomaly was found in 18/54 (33.3%) cases, therefore, overall FISH identified 75 anomalies of which 32 (42.6%) were undetected by CC. FISH provided additional information in cases with CC failure and in cases with a normal karyotype. Further, in ten cases with an abnormal karyotype, FISH could identify additional anomalies, increasing the number of abnormalities per patient. Although CC is the gold standard in the cytogenetic profiling of MDS, FISH has proven to be an asset in identifying additional abnormalities. The number of anomalies per patient can predict the prognosis in MDS and hence, FISH contributed towards prognostic re-categorization. The FISH Panel testing should be used as an adjunct to CC, irrespective of the adequacy of the number of metaphases in CC, as it improves the prognostic classification of MDS. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Improvement of automatic fish feeder machine design

    Science.gov (United States)

    Chui Wei, How; Salleh, S. M.; Ezree, Abdullah Mohd; Zaman, I.; Hatta, M. H.; Zain, B. A. Md; Mahzan, S.; Rahman, M. N. A.; Mahmud, W. A. W.

    2017-10-01

    Nation Plan of action for management of fishing is target to achieve an efficient, equitable and transparent management of fishing capacity in marine capture fisheries by 2018. However, several factors influence the fishery production and efficiency of marine system such as automatic fish feeder machine could be taken in consideration. Two latest fish feeder machines have been chosen as the reference for this study. Based on the observation, it has found that the both machine was made with heavy structure, low water and temperature resistance materials. This research’s objective is to develop the automatic feeder machine to increase the efficiency of fish feeding. The experiment has conducted to testing the new design of machine. The new machine with maximum storage of 5 kg and functioning with two DC motors. This machine able to distribute 500 grams of pellets within 90 seconds and longest distance of 4.7 meter. The higher speed could reduce time needed and increase the distance as well. The minimum speed range for both motor is 110 and 120 with same full speed range of 255.

  11. Anchoring novel molecular biomarker responses to traditional responses in fish exposed to environmental contamination

    International Nuclear Information System (INIS)

    Nogueira, Patricia; Pacheco, Mario; Lourdes Pereira, M.; Mendo, Sonia; Rotchell, Jeanette M.

    2010-01-01

    The responses of Dicentrarchus labrax and Liza aurata to aquatic pollution were assessed in a contaminated coastal lagoon, using both traditional and novel biomarkers combined. DNA damage, assessed by comet assay, was higher in both fish species from the contaminated sites, whereas levels of cytochrome P450 1A1 gene expression were not significantly altered. The liver histopathological analysis also revealed significant lesions in fish from contaminated sites. Alterations in ras and xpf genes were analysed and additional pollutant-responsive genes were identified. While no alterations were found in ras gene, a downregulation of xpf gene was observed in D. labrax from a contaminated site. Suppression subtractive hybridization applied to D. labrax collected at a contaminated site, revealed altered expression in genes involved in energy metabolism, immune system activity and antioxidant response. The approach and results reported herein demonstrate the utility of anchoring traditional biomarker responses alongside novel biomarker responses. - Novel molecular biomarkers of aquatic environmental contamination in fish.

  12. A combined RT-PCR and dot-blot hybridization method reveals the coexistence of SJNNV and RGNNV betanodavirus genotypes in wild meagre (Argyrosomus regius).

    Science.gov (United States)

    Lopez-Jimena, B; Cherif, N; Garcia-Rosado, E; Infante, C; Cano, I; Castro, D; Hammami, S; Borrego, J J; Alonso, M C

    2010-10-01

    To detect the possible coexistence of striped jack nervous necrosis virus (SJNNV) and red-spotted grouper nervous necrosis virus (RGNNV) genotypes in a single fish, a methodology based on the combination of PCR amplification and blot hybridization has been developed and applied in this study. The degenerate primers designed for the PCR procedure target the T4 region within the capsid gene, resulting in the amplification of both genotypes. The subsequent hybridization of these amplification products with two different specific digoxigenin-labelled probes resulted in the identification of both genotypes separately. The application of the RT-PCR protocol to analyse blood samples from asymptomatic wild meagre (Argyrosomus regius) specimens has shown a 46.87% of viral nervous necrosis virus carriers. The combination of RT-PCR and blot hybridization increases the detection rate up to 90.62%, and, in addition, it has shown the coexistence of both genotypes in 18 out of the 32 specimens analysed (56.25%). This study reports the coexistence of betanodaviruses belonging to two different genotypes (SJNNV and RGNNV) in wild fish specimens. This is the first report demonstrating the presence of SJNNV and RGNNV genotypes in the same specimen. This study also demonstrates a carrier state in this fish species for the first time. © 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.

  13. Analysis and design of hybrid control systems

    Energy Technology Data Exchange (ETDEWEB)

    Malmborg, J.

    1998-05-01

    Different aspects of hybrid control systems are treated: analysis, simulation, design and implementation. A systematic methodology using extended Lyapunov theory for design of hybrid systems is developed. The methodology is based on conventional control designs in separate regions together with a switching strategy. Dynamics are not well defined if the control design methods lead to fast mode switching. The dynamics depend on the salient features of the implementation of the mode switches. A theorem for the stability of second order switching together with the resulting dynamics is derived. The dynamics on an intersection of two sliding sets are defined for two relays working on different time scales. The current simulation packages have problems modeling and simulating hybrid systems. It is shown how fast mode switches can be found before or during simulation. The necessary analysis work is a very small overhead for a modern simulation tool. To get some experience from practical problems with hybrid control the switching strategy is implemented in two different software environments. In one of them a time-optimal controller is added to an existing PID controller on a commercial control system. Successful experiments with this hybrid controller shows the practical use of the method 78 refs, 51 figs, 2 tabs

  14. Effects of bamboo substrate and supplemental feeding on growth and production of hybrid red tilapia fingerlings (Oreochromis mossambicusxOrechromis niloticus)

    NARCIS (Netherlands)

    Keshavanath, P.; Gangadhar, B.; Ramesh, T.J.; Dam, van A.A.; Beveridge, M.C.M.; Verdegem, M.C.J.

    2004-01-01

    Periphyton growing on artificial substrates can increase the production of herbivorous fish in aquaculture ponds. Periphyton may be an alternative or a complement for supplemental feed in fingerling production. Growth and production of hybrid red tilapia (Oreochromis mossambicus x Oreochromis

  15. The effect of radiation on the storage of fish

    International Nuclear Information System (INIS)

    Ghadi, S.V.; Madhavan, V.N.; Kumta, U.S.; Lewis, N.F.

    1976-01-01

    Fresh fish spoils rapidly at +- 0 0 to + 2 0 C, and produces considerable quantities of ammonia. It is known that ammonia production is reduced if fish is heated before processing. Radiation experiments were carried out to see whether the storage stability of fresh fish could be improved without cooking it. Radiation with 100 to 500 Krad or steam cooking for 3 to 5 minutes did not improve storage stability. Only combined treatment with steam and low doses (100 Krad) of gamma rays lengthened refrigerated storage life. (orig.) [de

  16. Development of a new CARD-FISH protocol for quantification of Legionella pneumophila and its application in two hospital cooling towers.

    Science.gov (United States)

    Kirschner, A K T; Rameder, A; Schrammel, B; Indra, A; Farnleitner, A H; Sommer, R

    2012-06-01

    Open cooling towers are frequent sources of infections with Legionella pneumophila. The gold standard for the detection of Leg. pneumophila is based on cultivation lasting up to 10 days and detecting only culturable cells. Alternative fluorescence in situ hybridization (FISH) protocols have been proposed, but they result in faint fluorescence signals and lack specificity because of cross-hybridization with other Legionella species. Our aim was thus to develop a new FISH protocol for rapid and specific detection of Leg. pneumophila in water samples. A novel catalysed reporter deposition FISH (CARD-FISH) protocol for the detection of Leg. pneumophila was developed, which significantly enhanced signal intensity as well as specificity of the probe through the use of a novel competitor probe. The developed protocol was compared with the culture method for monitoring the seasonal development of culturable and nonculturable Leg. pneumophila in two hospital cooling tower systems. Seasonal fluctuations of Leg. pneumophila concentrations detected via CARD-FISH were related to the development of the total bacterial community in both cooling towers, with temperature and biocide as the main factors controlling this development. Our results clearly showed that the majority of the Leg. pneumophila cells were in a nonculturable state. Thus, detection of Leg. pneumophila with culture methods may underestimate the total numbers of Leg. pneumophila present. Rapid, sensitive and specific detection and quantification of Leg. pneumophila in water systems is prerequisite for reliable risk estimation. The new protocol significantly improves current methodology and can be used to monitor and screen for Leg. pneumophila concentrations in cooling towers or other water systems. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

  17. Association of Serpulina hyodysenteriae with the colonic mucosa in experimental swine dysentery studied by fluorescent in situ hybridization

    DEFF Research Database (Denmark)

    Jensen, Tim Kåre; Boye, Mette; Møller, Kristian

    1998-01-01

    The localization of Serpulina hyodysenteriae in experimental swine dysentery was studied by fluorescent in situ hybridization (FISH) using an oligonucleotide probe targeting the 23S rRNA of S. hyodysenteriae. Nine 8-week-old pigs were challenged. Seven of the pigs were intragastrically dosed with 1......x10(9) cfu S. hyodysenteriae for 3 consecutive days, whereas two pigs were infected by contact. Six non-challenged pigs served as negative controls. The challenged pigs developed clinical swine dysentery from 8 to 14 days postinfection with typical gross lesions. By FISH S. hyodysenteriae cells...

  18. Do bacteria, not fish, produce 'fish kairomone'?

    NARCIS (Netherlands)

    Ringelberg, J.; Van Gool, E.

    1998-01-01

    Fish-associated chemicals enhance phototactic downward swimming in Daphnia. If perch were treated with the antibiotic ampicillin, this enhancement was significantly decreased. Therefore, not fish, but bacteria associated with fish, seem to produce this kairomone. [KEYWORDS: Diel vertical migration;

  19. CHARACTERIZATION AND AUTOMATIC COUNTING OF F.I.S.H. SIGNALS IN 3-D TISSUE IMAGES

    Directory of Open Access Journals (Sweden)

    Umesh PS Adiga

    2011-05-01

    Full Text Available The evaluation of malignancy-related features often helps to determine the prognoses for patients with carcinomas. One technique, which is becoming increasingly important for assessing such prognostic features is that of Fluorescence in situ Hybridization (FISH. By counting the number of FISH signals in a stack of 2- D images of a tumor (which together constitute the 3-D image volume, it is possible to determine whether there has been any loss or gain of the target DNA sequences and thereby evaluate the stage of the disease. However, visual counting of the FISH signals in this way is a tedious, fatiguing and time-consuming task. Therefore, we have developed an automated system for the quantitative evaluation of FISH signals. We present and discuss the implementation of an image processing module that segments, characterizes and counts the FISH signals in 3-D images of thick prostate tumor tissue specimens. Possible errors in the automatic counting of signals are listed and ways to circumvent these errors are described. We define a feature vector for a FISH signal and describe how we have used the weighted feature vector to segment specific signals from noise artifacts. In addition, we present a method, which allows overlapping FISH signals to be distinguished by fitting a local Gaussian model around the intensity profile and studying the feature vector of each model. Our complete image processing module overcomes the problems of manual counting of FISH signals in 3-D images of tumor specimens, thereby providing improved diagnostic and prognostic capability in qualitative diagnostic pathology.

  20. Osmoregulatory effects of hypophysectomy and homologous prolactin replacement in hybrid striped bass

    DEFF Research Database (Denmark)

    Jackson, Leslie F; McCormick, Stephen D; Madsen, Steffen S

    2005-01-01

    The effects of ovine prolactin (oPRL) and striped bass prolactin (sbPRL; Morone saxatilis) on plasma osmolality, electrolyte balance, and gill Na+,K+-ATPase activity were investigated in hypophysectomized (Hx), freshwater (FW)-acclimated, hybrid striped bass (M. saxatilis x Morone chrysops...... or 100 ng/g), or hormone vehicle (0.9% NaCl) at 48-h intervals (days 0, 2, 4, and 6) in FW and then sampled for blood plasma 24 h after the fourth injection (day 7). In Hx fish, oPRL (5 and 20 microg/g) and sbPRL (10 and 100 ng/g) were effective in maintaining plasma osmolality and levels of Na+, Cl...... balance in FW-adapted hybrid striped bass, and that this may involve downregulation of branchial Na+,K+-ATPase activity....

  1. Karyotype and Mapping of Repetitive DNAs in the African Butterfly Fish Pantodon buchholzi, the Sole Species of the Family Pantodontidae

    Czech Academy of Sciences Publication Activity Database

    Ráb, Petr; Yano, C. F.; Lavoué, S.; Jegede, O. I.; Bertollo, L.A.C.; Ezaz, T.; Majtánová, Zuzana; de Oliveira, E. A.; Cioffi, M.B.

    2016-01-01

    Roč. 149, č. 4 (2016), s. 312-320 ISSN 1424-8581 R&D Projects: GA ČR GA14-02940S Institutional support: RVO:67985904 Keywords : African butterfly fish * conventional staining techniques * fluorescence in situ hybridization Subject RIV: EG - Zoology Impact factor: 1.354, year: 2016

  2. Chromosomal distribution of pTa-535, pTa-86, pTa-713, 35S rDNA repetitive sequences in interspecific hexaploid hybrids of common wheat (Triticum aestivum L. and spelt (Triticum spelta L..

    Directory of Open Access Journals (Sweden)

    Klaudia Goriewa-Duba

    Full Text Available Fluorescent in situ hybridization (FISH relies on fluorescent-labeled probes to detect specific DNA sequences in the genome, and it is widely used in cytogenetic analyses. The aim of this study was to determine the karyotype of T. aestivum and T. spelta hybrids and their parental components (three common wheat cultivars and five spelt breeding lines, to identify chromosomal aberrations in the evaluated wheat lines, and to analyze the distribution of polymorphisms of repetitive sequences in the examined hybrids. The FISH procedure was carried out with four DNA clones, pTa-86, pTa-535, pTa-713 and 35S rDNA used as probes. The observed polymorphisms between the investigated lines of common wheat, spelt and their hybrids was relatively low. However, differences were observed in the distribution of repetitive sequences on chromosomes 4A, 6A, 1B and 6B in selected hybrid genomes. The polymorphisms observed in common wheat and spelt hybrids carry valuable information for wheat breeders. The results of our study are also a valuable source of knowledge about genome organization and diversification in common wheat, spelt and their hybrids. The relevant information is essential for common wheat breeders, and it can contribute to breeding programs aimed at biodiversity preservation.

  3. Chromosomal distribution of pTa-535, pTa-86, pTa-713, 35S rDNA repetitive sequences in interspecific hexaploid hybrids of common wheat (Triticum aestivum L.) and spelt (Triticum spelta L.).

    Science.gov (United States)

    Goriewa-Duba, Klaudia; Duba, Adrian; Kwiatek, Michał; Wiśniewska, Halina; Wachowska, Urszula; Wiwart, Marian

    2018-01-01

    Fluorescent in situ hybridization (FISH) relies on fluorescent-labeled probes to detect specific DNA sequences in the genome, and it is widely used in cytogenetic analyses. The aim of this study was to determine the karyotype of T. aestivum and T. spelta hybrids and their parental components (three common wheat cultivars and five spelt breeding lines), to identify chromosomal aberrations in the evaluated wheat lines, and to analyze the distribution of polymorphisms of repetitive sequences in the examined hybrids. The FISH procedure was carried out with four DNA clones, pTa-86, pTa-535, pTa-713 and 35S rDNA used as probes. The observed polymorphisms between the investigated lines of common wheat, spelt and their hybrids was relatively low. However, differences were observed in the distribution of repetitive sequences on chromosomes 4A, 6A, 1B and 6B in selected hybrid genomes. The polymorphisms observed in common wheat and spelt hybrids carry valuable information for wheat breeders. The results of our study are also a valuable source of knowledge about genome organization and diversification in common wheat, spelt and their hybrids. The relevant information is essential for common wheat breeders, and it can contribute to breeding programs aimed at biodiversity preservation.

  4. Adoption Of Improved Fish Technologies Among Fish Farmers In ...

    African Journals Online (AJOL)

    A shortfall exists between fish supply and fish demand in the country despite the introduction of improved technology to fish farmers. This led to huge wage bill on the importation of fish to meet the protein need of the ever increasing population. This prompted this study with focus on adoption of improved fish technologies ...

  5. Automated DNA electrophoresis, hybridization and detection

    International Nuclear Information System (INIS)

    Zapolski, E.J.; Gersten, D.M.; Golab, T.J.; Ledley, R.S.

    1986-01-01

    A fully automated, computer controlled system for nucleic acid hybridization analysis has been devised and constructed. In practice, DNA is digested with restriction endonuclease enzyme(s) and loaded into the system by pipette; 32 P-labelled nucleic acid probe(s) is loaded into the nine hybridization chambers. Instructions for all the steps in the automated process are specified by answering questions that appear on the computer screen at the start of the experiment. Subsequent steps are performed automatically. The system performs horizontal electrophoresis in agarose gel, fixed the fragments to a solid phase matrix, denatures, neutralizes, prehybridizes, hybridizes, washes, dries and detects the radioactivity according to the specifications given by the operator. The results, printed out at the end, give the positions on the matrix to which radioactivity remains hybridized following stringent washing

  6. Two-dimensional Semiconductor-Superconductor Hybrids

    DEFF Research Database (Denmark)

    Suominen, Henri Juhani

    This thesis investigates hybrid two-dimensional semiconductor-superconductor (Sm-S) devices and presents a new material platform exhibiting intimate Sm-S coupling straight out of the box. Starting with the conventional approach, we investigate coupling superconductors to buried quantum well....... To overcome these issues we integrate the superconductor directly into the semiconducting material growth stack, depositing it in-situ in a molecular beam epitaxy system under high vacuum. We present a number of experiments on these hybrid heterostructures, demonstrating near unity interface transparency...

  7. Maggot meal as a substitute for fish meal in laying chicken diet ...

    African Journals Online (AJOL)

    A 56-day experiment was conducted to determine thereplacement value of maggot meal for fish meal in diet of laying chicken. Fish meal was included at 30 g kg-1 of the control diet. Maggot meal incrementally replaced fish meal at 250, 500, 750, and 1000 g kg-1 on crude protein basis. The five diets were fed to a total of ...

  8. Which Fish Should I Eat? Perspectives Influencing Fish Consumption Choices

    Science.gov (United States)

    Choi, Anna L.; Karagas, Margaret R.; Mariën, Koenraad; Rheinberger, Christoph M.; Schoeny, Rita; Sunderland, Elsie; Korrick, Susan

    2012-01-01

    Background: Diverse perspectives have influenced fish consumption choices. Objectives: We summarized the issue of fish consumption choice from toxicological, nutritional, ecological, and economic points of view; identified areas of overlap and disagreement among these viewpoints; and reviewed effects of previous fish consumption advisories. Methods: We reviewed published scientific literature, public health guidelines, and advisories related to fish consumption, focusing on advisories targeted at U.S. populations. However, our conclusions apply to groups having similar fish consumption patterns. Discussion: There are many possible combinations of matters related to fish consumption, but few, if any, fish consumption patterns optimize all domains. Fish provides a rich source of protein and other nutrients, but because of contamination by methylmercury and other toxicants, higher fish intake often leads to greater toxicant exposure. Furthermore, stocks of wild fish are not adequate to meet the nutrient demands of the growing world population, and fish consumption choices also have a broad economic impact on the fishing industry. Most guidance does not account for ecological and economic impacts of different fish consumption choices. Conclusion: Despite the relative lack of information integrating the health, ecological, and economic impacts of different fish choices, clear and simple guidance is necessary to effect desired changes. Thus, more comprehensive advice can be developed to describe the multiple impacts of fish consumption. In addition, policy and fishery management inter-ventions will be necessary to ensure long-term availability of fish as an important source of human nutrition. PMID:22534056

  9. HER-2 protein concentrations in breast cancer cells increase before immunohistochemical and fluorescence in situ hybridization analysis turn positive

    DEFF Research Database (Denmark)

    Olsen, Dorte A; Østergaard, Birthe; Bokmand, Susanne

    2007-01-01

    BACKGROUND: The level of HER-2/neu in breast cancer cells is normally measured by immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH). It determines whether patients should be treated with trastuzumab (Herceptin). In this study, HER-2 protein in breast cancer tissue...

  10. Fish Rhabdoviruses

    Science.gov (United States)

    Kurath, G.; Winton, J.

    2008-01-01

    Many important viral pathogens of fish are members of the family Rhabdoviridae. The viruses in this large group cause significant losses in populations of wild fish as well as among fish reared in aquaculture. Fish rhabdoviruses often have a wide host and geographic range, and infect aquatic animals in both freshwater and seawater. The fish rhabdoviruses comprise a diverse collection of isolates that can be placed in one of two quite different groups: isolates that are members of the established genusNovirhabdovirus, and those that are most similar to members of the genus Vesiculovirus. Because the diseases caused by fish rhabdoviruses are important to aquaculture, diagnostic methods for their detection and identification are well established. In addition to regulations designed to reduce the spread of fish viruses, a significant body of research has addressed methods for the control or prevention of diseases caused by fish rhabdoviruses, including vaccination. The number of reported fish rhabdoviruses continues to grow as a result of the expansion of aquaculture, the increase in global trade, the development of improved diagnostic methods, and heightened surveillance activities. Fish rhabdoviruses serve as useful components of model systems to study vertebrate virus disease, epidemiology, and immunology.

  11. Fish for Feed vs Fish for Food

    OpenAIRE

    Allan, Geoff L.

    2004-01-01

    Aquaculture is the fastest-growing food producing industry sector in the world. Demand for feed ingredients, particularly for preferred protein sources such as fishmeal, fish oil and ‘trash fish’, has also increased, raising questions about sustainability and uses of fish for aquaculture feeds or directly as human food. Approximately 30 million metric tonnes (MMT) of fish from capture fisheries are used each year to produce fishmeal and fish oil. The species used are not usually consumed dire...

  12. Fish Allergy

    Science.gov (United States)

    ... Safe Videos for Educators Search English Español Fish Allergy KidsHealth / For Parents / Fish Allergy What's in this ... Print en español Alergia al pescado About Fish Allergy A fish allergy is not exactly the same ...

  13. GeoNeoLogical novel and other hybrimedia experiments: or how to use hybrid methods such as Hybrid Discourse Analysis (HDA within a knowledge base

    Directory of Open Access Journals (Sweden)

    Andrade, Pedro

    2016-07-01

    Full Text Available In this essay I will present some results of the project Public Communication of Art, which developed a seminal theory and methodology intended to cope with hybridity and new media literacy in our globalized and inter/transcultural world. Some of the methods used blend vision with touch and are called ‘hybrid methods’ or ‘hybrimethods’. Examples of these are, for instance, a Multitouch Interactive Table, a Multitouch Questionnaire, Trichotomies Game and GeoNeoLogic Novel, this last one being a hybrid novel activated by fusion of vision, touch and GPS coordinates. Another hybrimethod is a sort of discursive analysis, named Hybrid Discourse Analysis (HDA, which uses ‘semantic-logical networks’ organized by concepts and ‘relation-concepts’. HDA is here articulated with Critical Sociology and applied to the analysis of a text on Magic Realism, which is also a hybrid genre within the social field of literature.

  14. Effect of pile-driving sounds on the survival of larval fish

    NARCIS (Netherlands)

    Bolle, L.J.; Jong, C.A.F. de; Bierman, S.M.; Beek, P.J.G. van van; Wessels, P.W.; Blom, E.; Damme, C.J.G. van; Winter, H.V.; Dekeling, R.P.A.

    2016-01-01

    Concern exists about the potential effects of pile-driving sounds on fish, but evidence is limited, especially for fish larvae. A device was developed to expose larvae to accurately reproduced pile-driving sounds. Controlled exposure experiments were carried out to examine the lethal effects in

  15. The hybrid electric vehicle revolution, off road

    Energy Technology Data Exchange (ETDEWEB)

    Wood, B.E. [ePower Technologies (United States)

    2004-07-01

    In this presentation the author presents concepts and details of hybrid vehicles in general, including their benefits, then describes off-road hybrid vehicles. Hybrid vehicles have been experimented with for over a century. Demonstrator vehicles include a diesel-electric tractor, an electric lawn tractor, a hybrid snow thrower, and a hybrid wheel loader. A duty cycle for the loader is shown with battery-assisted acceleration, and regenerative braking. Both of these keep the size of the engine small, the loads on it less variable, thus improving fuel economy. A hybrid excavator and its duty cycle is shown. A fuel cell lift truck that is currently in design is illustrated. The author then describes the possibilities of the hydrogen economy where sourcing and infrastructure are yet to be demonstrated on a commercial scale. The author predicts that off-road hydrogen fuel cell vehicles will be commercially viable five years before on-road applications. The author predicts hydrogen sourced from biogas, photovoltaics, and wind power. tabs, figs.

  16. State-of-the-art assessment of electric and hybrid vehicles

    Science.gov (United States)

    1978-01-01

    Data are presented that were obtained from the electric and hybrid vehicles tested, information collected from users of electric vehicles, and data and information on electric and hybrid vehicles obtained on a worldwide basis from manufacturers and available literature. The data given include: (1) information and data base (electric and hybrid vehicle systems descriptions, sources of vehicle data and information, and sources of component data); (2) electric vehicles (theoretical background, electric vehicle track tests, user experience, literature data, and summary of electric vehicle status); (3) electric vehicle components (tires, differentials, transmissions, traction motors, controllers, batteries, battery chargers, and component summary); and (4) hybrid vehicles (types of hybrid vehicles, operating modes, hybrid vehicles components, and hybrid vehicles performance characteristics).

  17. Safety of oxytetracycline (Terramycin TM-100F) administered in feed to hybrid striped bass, walleyes, and yellow perch

    Science.gov (United States)

    Gaikowski, M.P.; Wolf, J.C.; Schleis, S.M.; Gingerich, W.H.

    2003-01-01

    Oxytetracycline (Terramycin TM-100F, a medicated premix containing oxytetracycline at 220 g/kg) is approved in the United States to control certain systemic bacterial diseases of salmon and catfish when fed at a rate of 55-82.5 mg per kilogram of bodyweight per day for 10 d. Although oxytetracycline may also control certain systemic bacterial infections in coolwater or scaled warmwater fish, no safety data for such species are available. Our objective was to determine the safety of oxytetracycline administered in feed at nominal doses of 0, 82.5, 248, or 413 mg??kg-1??d-1 to yellow perch Perca flavescens and hybrid striped bass (striped bass Morone saxatilis x white bass M. chrysops) for 10 d and to walleye Sander vitreus (formerly Stizostedion vitreum) for 20 d. Yellow perch and hybrid striped bass consumed 50% to 100% of the diet, whereas walleye feed consumption was occasionally less than 50% of the diet. Feed or fecal material was present in the gastrointestinal tract of all necropsied walleyes except for one control fish. The single growth effect was that hybrid striped bass offered a nominal dose of 413 mg??kg-1??d-1 were significantly smaller than untreated controls. Oxytetracycline-related histopathological findings were limited to walleyes and were of low severity. The histopathological findings included decreased hematopoietic-lymphopoietic (H&L) tissue in the anterior kidneys, diffuse hyperplasia of the gill filament epithelium, and a decreased prevalence of fish with eosinophilic droplets in their renal tubular epithelial cells. Although the incidence of decreased H&L tissue tended to increase in proportion to oxytetracycline dose, this finding was statistically significant only for fish that received a nominal dose of 413 mg??kg-1??d-1. Given the pathogenicity of the types of bacteria that are controlled by oxytetracycline treatment and the long history of its use in major aquaculture species, the relative risk of the minor oxytetracycline

  18. Effect of temperature on spoilage of fresh and frozen fish sold in ...

    African Journals Online (AJOL)

    African Journal of Livestock Extension ... The TMA value recorded in live-fish at the start of the experiment (2.2 mg TMA/100g ... Safe presentation and marketing practices and establishment of fish inspection services is hereby recommended.

  19. Changes in the fish fauna of the Kissimmee River basin, peninsular Florida: Nonnative additions

    Science.gov (United States)

    Nico, L.G.

    2005-01-01

    Recent decades have seen substantial changes in fish assemblages in rivers of peninsular Florida. The most striking change has involved the addition of nonnative fishes, including taxa from Asia, Africa, and Central and South America. I review recent and historical records of fishes occurring in the Kissimmee River basin (7,800 km2), a low-gradient drainage with 47 extant native fishes (one possibly the result of an early transplant), at least 7 foreign fishes (most of which are widely established), and a stocked hybrid. Kissimmee assemblages include fewer marine fishes than the nearby Peace and Caloosahatchee rivers, and fewer introduced foreign fishes than south Florida canals. Fish assemblages of the Kissimmee and other subtropical Florida rivers are dynamic, due to new introductions, range expansions of nonnative fishes already present, and periodic declines in nonnative fish populations during occasional harsh winters. The addition, dispersal, and abundance of nonnative fishes in the basin is linked to many factors, including habitat disturbance, a subtropical climate, and the fact that the basin is centrally located in a region where drainage boundaries are blurred and introductions of foreign fishes commonplace. The first appearance of foreign fishes in the basin coincided with the complete channelization of the Kissimmee River in the 1970s. Although not a causal factor, artificial waterways connecting the upper lakes and channelization of the Kissimmee River have facilitated dispersal. With one possible exception, there have been no basin-wide losses of native fishes. When assessing change in peninsular Florida waters, extinction or extirpation of fishes appears to be a poor measure of impact. No endemic species are known from peninsular Florida (although some endemic subspecies have been noted). Most native freshwater fishes are themselves descended from recent invaders that reached the peninsula from the main continent. These invasions likely were

  20. Chromosome mapping by FISH to metaphase and interphase nuclei. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Trask, B.

    1997-08-01

    The overall specific aims of this project were: (1) to determine the large-scale structure of interphase and metaphase chromosomes, in order to establish new capabilities for genome mapping by fluorescence in situ hybridization (FISH); (2) to detect chromosome abnormalities associated with genetic disease and map DNA sequences relative to them in order to facilitate the identification of new genes with disease-causing mutations; (3) to establish medium resolution physical maps of selected chromosomal regions using a combined metaphase and interphase mapping strategy and to corroborate physical and genetic maps and integrate these maps with the cytogenetic map; (4) to analyze the polymorphism and sequence evolution of subtelomeric regions of human chromosomes; (5) to establish a state-of-the-art FISH and image processing facility in the Department of Molecular Biotechnology, University of Washington, in order to map DNA sequences rapidly and accurately to benefit the Human Genome Project.

  1. Genetic Relationships among Hylocereus and Selenicereus Vine Cacti (Cactaceae): Evidence from Hybridization and Cytological Studies

    Science.gov (United States)

    TEL-ZUR, NOEMI; ABBO, SHAHAL; BAR-ZVI, DUDY; MIZRAHI, YOSEF

    2004-01-01

    • Background and Aims Hylocereus and Selenicereus are native to tropical and sub-tropical America. Based on its taxonomic status and crossability relations it was postulated that H. megalanthus (syn. S. megalanthus) is an allotetraploid (2n = 4x = 44) derived from natural hybridization between two closely related diploid taxa. The present work aimed at elucidating the genetic relationships between species of the two genera. • Methods Crosses were performed and the putative hybrids were analysed by chromosome counts and morphological traits. The ploidy level of hybrids was confirmed by fluorescent in situ hybridization (FISH) of rDNA sites. Genomic in situ hybridization (GISH) was used in an attempt to identify the putative diploid genome donors of H. megalanthus and an artificial interploid hybrid. • Key Results Reciprocal crosses among four diploid Hylocereus species (H. costaricensis, H. monacanthus (syn. H. polyrhizus), H. undatus and Hylocereus sp.) yielded viable diploid hybrids, with regular chromosome pairing. Reciprocal crosses between these Hylocereus spp. and H. megalanthus yielded viable triploid, pentaploid, hexaploid and aneuploid hybrids. Morphological and phenological traits confirm the hybrid origin. In situ detection of rDNA sites was in accord with the ploidy status of the species and hybrid studied. GISH results indicated that overall sequence composition of H. megalanthus is similar to that of H. ocamponis and S. grandiflorus. High sequence similarity was also found between the parental genomes of H. monacanthus and H. megalanthus in one triploid hybrid. • Conclusions The ease of obtaining partially fertile F1 hybrids and the relative sequence similarity (in GISH study) suggest close genetic relationships among the taxa analysed. PMID:15329334

  2. Catch shares slow the race to fish

    Science.gov (United States)

    Birkenbach, Anna M.; Kaczan, David J.; Smith, Martin D.

    2017-04-01

    In fisheries, the tragedy of the commons manifests as a competitive race to fish that compresses fishing seasons, resulting in ecological damage, economic waste, and occupational hazards. Catch shares are hypothesized to halt the race by securing each individual’s right to a portion of the total catch, but there is evidence for this from selected examples only. Here we systematically analyse natural experiments to test whether catch shares reduce racing in 39 US fisheries. We compare each fishery treated with catch shares to an individually matched control before and after the policy change. We estimate an average policy treatment effect in a pooled model and in a meta-analysis that combines separate estimates for each treatment-control pair. Consistent with the theory that market-based management ends the race to fish, we find strong evidence that catch shares extend fishing seasons. This evidence informs the current debate over expanding the use of market-based regulation to other fisheries.

  3. Hybrid carrageenans: isolation, chemical structure, and gel properties.

    Science.gov (United States)

    Hilliou, Loic

    2014-01-01

    Hybrid carrageenan is a special class of carrageenan with niche application in food industry. This polysaccharide is extracted from specific species of seaweeds belonging to the Gigartinales order. This chapter focuses on hybrid carrageenan showing the ability to form gels in water, which is known in the food industry as weak kappa or kappa-2 carrageenan. After introducing the general chemical structure defining hybrid carrageenan, the isolation of the polysaccharide will be discussed focusing on the interplay between seaweed species, extraction parameters, and the hybrid carrageenan chemistry. Then, the rheological experiments used to determine the small and large deformation behavior of gels will be detailed before reviewing the relationships between gel properties and hybrid carrageenan chemistry. © 2014 Elsevier Inc. All rights reserved.

  4. Monitoring stress in fish by applying image analysis to their skin mucous cells

    Directory of Open Access Journals (Sweden)

    I. N. Vatsos

    2010-05-01

    Full Text Available Several authors have previously demonstrated that the number of the skin mucous cells of fish is affected by many stressors. In the present study, two experiments were conducted in order to examine the effects of two common environmental conditions on the morphology of skin of sea bass and particularly on the number and diameter of skin mucous cells. In the first experiment, two groups of sea bass (mean weight 155.6±10.3 g SD were maintained in two different concentrations of nitrate, 100 and 700 ppm respectively, for 48 h, while a third group was used as control. In the second experiment, sea bass (initial mean weight 78.9±3.1 g SD were divided into four groups and each group was maintained in a different level of oxygen for 9 weeks. The oxygen concentration in each group was: 3.6±0.2 ppm, 4.7±0.2 ppm, 6.2±0.2 ppm and 8.2±0.2 ppm. In both experiments the effects of the two environmental factors on the morphology of the fish skin were examined histologically and a software containing a visual basic script macro, allowing quantification of the skin mucous cells, was used to analyze the skin tissue sections. Concerning the overall morphology of the skin and the diameter of the skin mucous cells, no differences were noted in both experiments (P>0.05. It was demonstrated however, that fish maintained in the lowest oxygen level and fish maintained in the highest concentration of nitrate exhibited significantly increased number of mucous cells per skin area (mm2. There is evidence that the enumeration of the skin mucous cells of fish can be used to monitor stress in fish.

  5. Fluorescence in situ hybridization: an improved method of quantitating chromosome damage and repair

    International Nuclear Information System (INIS)

    Brown, J.M.; Evans, J.W.

    1993-01-01

    The authors combined fluorescence in situ hybridization (FISH) with specific full-length chromosome probes using the premature chromosome condensation (PCC) technique chromosome condensation (PCC) technique to simplify scoring chromosome damage and its repair. They have shown the technique works well and enables breaks and exchanges to be readily detected and scored in individual chromosomes. A chromosome 4 full-length specific library has been used in initial studies. (UK)

  6. Model identification and controller design of a fish-like robot

    Science.gov (United States)

    Ariyanto, Irfan; Kang, Taesam; Chan, Wai Leung; Lee, Youngjae

    2007-04-01

    Robotic fish is an interesting and prospective subject to develop. The simplest fish swimming mode to be mimicked for fish robots is the ostraciiform mode which only requires caudal fin flapping. An almost submerged ostraciiform fish robot was constructed to study its swimming characteristics. The swimming direction can be controlled by changing the mean angle of caudal fin oscillation. Experiments were conducted to study the behavior of the fish robot and in particular, the transfer function between swimming path angular rate and mean angle of the caudal fin oscillation were identified. Error to signal ratio quantity was used to determine how well the model fits with the experimental data. This identification model was used to design a 2-degree-of-freedom PID controller that meets some specific requirements to improve the steering performance.

  7. "Flat-Fish" vacuum chamber for the ISR

    CERN Multimedia

    CERN PhotoLab

    1983-01-01

    The "Flat-Fish" vacuum chamber was a very special one for the ISR beam intersection I-4, inside the Split-Field Magnet. Technical descriptions are given with pictures 7802042 and 7712182. There were several generations of this chamber. This picture was taken during the last year of ISR operation. See also 7510230X, 7511123X, 7802039, 7802041. A flat-fish type vacuum chamber was also used at I-8, inside the Open Axial Field Magnet (OAFM, experiment 807 on large transverse momentum collision products), see 7901358.

  8. Construction of radiation-reduced hybrids and their use in mapping of microclones from chromosome 10p11.2-q11.2

    International Nuclear Information System (INIS)

    Fujita, Shoichi; Shin, Eisei; Nakamura, Tsutomu; Kurahashi, Hiroki; Mori, Takesada; Takai, Shin-ichiro; Nishisho, Isamu; Kaneda, Yasufumi; Tanaka, Kiyoji.

    1993-01-01

    Radiation-reduced hybrids for mapping of DNA markers in the pericentromeric region of chromosome 10 were developed. A Chinese hamster/human somatic cell hybrid (762-8A) carrying chromosomes 10 and Y as the only human material were exposed to 40,000 rads of irradiation and then rescued by fusion with non-irradiated recipient Chinese hamster cells (GM459). Southern hybridization analyses revealed that 10 of 128 HAT-resistant clones contained human chromosomal fragments corresponding to at least one marker locus between FNRB (10p11.2) and RBP3 (10q11.2). These hybrids were then used to map microdissection clones previously isolated and roughly mapped to this chromosomal region by fluorescence in situ hybridization (FISH). Two of the six microclones studies could be mapped to the proximity of the D10S102 locus. These radiation hybrids are useful for the construction of refined genetic maps of the pericentromeric region of chromosome 10. (author) 50 refs

  9. Fish consumption and track to a fish feed formulation

    Science.gov (United States)

    Cai-Juan, Soong; Ramli, Razamin; Rahman, Rosshairy Abdul

    2015-12-01

    Strategically located in the equator, Malaysia is blessed with plenty of fish supply. The high demand in fish consumption has helped the development in the fishery industry and provided numerous jobs in the secondary sector, contributing significantly to the nation's income. A survey was conducted to understand the trend of current demands for fish for the purpose of designing a feed formulation, which is still limited in this area of study. Results showed that grouper fish in restaurants commanded a very high price compared to other species of fish. Tiger grouper gained the highest demand in most restaurants, while giant grouper had the highest price in restaurants. Due to the demand and challenges to culture this type of fish, a framework for fish feed formulation is proposed. The formulation framework when materialized could be an alternative to the use of trash fish as the feed for grouper.

  10. 75 FR 6058 - Federal Sport Fish Restoration; California Department of Fish and Game Fish Hatchery and Stocking...

    Science.gov (United States)

    2010-02-05

    ...] Federal Sport Fish Restoration; California Department of Fish and Game Fish Hatchery and Stocking Program... (NEPA) of 1969, as amended, for the EIR/EIS jointly prepared with CDFG. Under the Sport Fish Restoration... has authority to grant Federal funds from the Sport Fish Restoration and Boating Trust Fund to support...

  11. Human cDNA mapping using fluorescence in situ hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Korenberg, J.R.

    1993-03-04

    Genetic mapping is approached using the techniques of high resolution fluorescence in situ hybridization (FISH). This technology and the results of its application are designed to rapidly generate whole genome as tool box of expressed sequence to speed the identification of human disease genes. The results of this study are intended to dovetail with and to link the results of existing technologies for creating backbone YAC and genetic maps. In the first eight months, this approach generated 60--80% of the expressed sequence map, the remainder expected to be derived through more long-term, labor-intensive, regional chromosomal gene searches or sequencing. The laboratory has made significant progress in the set-up phase, in mapping fetal and adult brain and other cDNAs, in testing a model system for directly linking genetic and physical maps using FISH with small fragments, in setting up a database, and in establishing the validity and throughput of the system.

  12. The effect of hybridization on fish physiology, immunity and blood biochemistry: A case study in hybridizing Cyprinus carpio and Carassius gibelio (Cyprinidae)

    Czech Academy of Sciences Publication Activity Database

    Šimková, A.; Vojtek, L.; Halačka, Karel; Hyršl, P.; Vetešník, Lukáš

    2015-01-01

    Roč. 435, č. 1 (2015), s. 381-389 ISSN 0044-8486 R&D Projects: GA ČR GAP505/12/0375 Institutional support: RVO:68081766 Keywords : Hybridization * Cyprinids * Blood biochemistry * Hematology * Immunity Subject RIV: EG - Zoology Impact factor: 1.893, year: 2015

  13. Modifikasi dan Uji Kinerja Alat Pengering Energi Surya-Hybrid Tipe Rak untuk Pengeringan Ikan Teri

    Directory of Open Access Journals (Sweden)

    Risman Hanafi

    2017-04-01

    Abstract. At the time of harvest fish, fishermen get a lot of anchovy as catches with very large amount. Due to the very large number of these, sometimes anchovy is not sold out. This resulted in an anchovy rot if not done preservation (cool storage. One way in which the fisherman is by drying anchovy naturally (the drying in the sun. A tool rack type solar dryer is a box-shaped dryer that utilize the sun as thermal energy. The constraints of this tool is only utilizing the heat from solar energy, so when the weather is cloudy or at nightfall these tools can not function. The research aims to modify the tool rack type solar dryer to be a hybrid dryer for drying anchovy. The results were obtained that total efficiency of energy use during drying is for drying empty test hybrid is 0,010%, for drying hybrid test at 0,695%, and for drying solar test of 20,319%. As for the length of drying time, hybrid test replicates 1 for 7 hours, hybrid test replicates 2 for 8 hours, solar test replicates 1 for 10 hours, and solar test replicates 2 for 11 hours. For the total energy available, hybrid drying of 305,838 MJ and solar drying of 9,896 MJ.

  14. Relative and combined effects of habitat and fishing on reef fish communities across a limited fishing gradient at Ningaloo.

    Science.gov (United States)

    Wilson, Shaun K; Babcock, Russ C; Fisher, Rebecca; Holmes, Thomas H; Moore, James A Y; Thomson, Damian P

    2012-10-01

    Habitat degradation and fishing are major drivers of temporal and spatial changes in fish communities. The independent effects of these drivers are well documented, but the relative importance and interaction between fishing and habitat shifts is poorly understood, particularly in complex systems such as coral reefs. To assess the combined and relative effects of fishing and habitat we examined the composition of fish communities on patch reefs across a gradient of high to low structural complexity in fished and unfished areas of the Ningaloo Marine Park, Western Australia. Biomass and species richness of fish were positively correlated with structural complexity of reefs and negatively related to macroalgal cover. Total abundance of fish was also positively related to structural complexity, however this relationship was stronger on fished reefs than those where fishing is prohibited. The interaction between habitat condition and fishing pressure is primarily due to the high abundance of small bodied planktivorous fish on fished reefs. However, the influence of management zones on the abundance and biomass of predators and target species is small, implying spatial differences in fishing pressure are low and unlikely to be driving this interaction. Our results emphasise the importance of habitat in structuring reef fish communities on coral reefs especially when gradients in fishing pressure are low. The influence of fishing effort on this relationship may however become more important as fishing pressure increases. Copyright © 2012 Elsevier Ltd. All rights reserved.

  15. Reformational direction of fish industry development in Ukraine

    Directory of Open Access Journals (Sweden)

    Yu. Glebova

    2017-12-01

    Full Text Available Purpose. Exploitation of aquatic bioresources in Ukraine was traditionally low. In order to provide the population of Ukraine with high-quality fish products, the task of fish industry workers in the country is to create appropriate conditions for increasing fish catches and improve the organization of labor and material provision at fish farms. Our immediate goal in the work was to identify the areas of reforms in the fish industry, which would ensure the progress in it and would be aimed at attracting investments in aquaculture, establishing fish protection bodies with a qualitative change in their working principles, forming a culture of socially responsible fishing, achieving transparency and openness in the work of the State Agency for Fisheries, as well as improving the efficiency of the functioning of state institutions. Methodology. The basis of the research is the scientific, statistical and reporting data in the fish industry of Ukraine. The research uses the methods of monitoring and retrospective analysis of recent research and publications. Findings. The information on the implementation of reforms in the field of fish industry in Ukraine is presented: change of territorial bodies, State Fisheries Agency, fisheries regimes, use of aquatic biological resources, regulatory framework, strengthening of international activities and use of foreign experience. In order to increase the capacity of the fish industry, it is being reformed in the direction of changing its structure, increasing funding, introducing foreign investment and using their expertise. Originality. The need for reforming the fish industry of Ukraine became highly important. Modern socio-economic conditions require changes in the approaches to the organization of the activity of f groups that are marked by significant technological, economic, organizational, regulatory and legal peculiarities that need to be taken into account in economic processes. In accordance with

  16. Tubular and endothelial chimerism in renal allografts using fluorescence and chromogenic in situ hybridization (FISH, CISH) technology.

    Science.gov (United States)

    Varga, Zsuzsanna; Gaspert, Ariana; Behnke, Silvia; von Teichman, Adriana; Fritzsche, Florian; Fehr, Thomas

    2012-04-01

    The role of endothelial and tubular chimerism in renal allograft adaptation and rejection varies in different studies. We addressed the correlation between different clinico-pathological settings and sex-chromosomal endothelial and/or tubular chimerism in renal allografts. We examined the presence or absence of the X and Y chromosomes by fluorescence and chromogenic in situ hybridization (FISH, CISH) methodology on paraffin embedded kidney biopsies in 16 gender mismatched renal transplants (1 to 12 years post-transplantation). Twelve patients were male, four female. Four groups were selected: (i) Vascular calcineurin inhibitor toxicity without rejection; (ii) T-cell mediated vascular rejection; (iii) antibody mediated rejection; and (iv) C4d-positivity in AB0-incompatible transplants with or without rejection. Twelve non-transplant kidney biopsies (8 female, 4 male) were used as controls. Tubular chimerism was detected more frequently (69%) than endothelial chimerism (12%) in renal transplants. One of 12 control patients had tubular and endothelial chimeric cells (8%). The Y chromosome occurred in 8/12 male recipients (67%) in tubular epithelial cells and in 5/12 male recipients (42%) in endothelial cells. Double X chromosomes were detected in 3/4 female recipients in tubular epithelium. Tubular chimerism occurred more often with endothelial chimerism and capillaritis without correlation with other parameters, such as rejection. Combined Y chromosomal tubular and lymphatic endothelial chimerism correlated with T-cell mediated vascular rejection in two out of three patients (66%). Combined Y chromosomal tubular and peritubular capillary chimerism correlated with antibody mediated C4d+ rejection in one out of two patients (50%). Tubular and/or endothelial chimerism occur frequently in gender mismatched renal allografts and, when combined, this is associated with T-cell mediated rejection. © 2012 The Authors. Pathology International © 2012 Japanese Society of

  17. Hydrodynamics of vertical jumping in Archer fish

    Science.gov (United States)

    Techet, Alexandra H.; Mendelson, Leah

    2017-11-01

    Vertical jumping for aerial prey from an aquatic environment requires both propulsive power and precise aim to succeed. Rapid acceleration to a ballistic velocity sufficient for reaching the prey height occurs before the fish leaves the water completely and experiences a thousandfold drop in force-producing ability. In addition to speed, accuracy and stability are crucial for successful feeding by jumping. This talk examines the physics of jumping using the archer fish as a model. Better known for their spitting abilities, archer fish will jump multiple body lengths out of the water for prey capture, from a stationary position just below the free surface. Modulation of oscillatory body kinematics and use of multiple fins for force production are identified as methods through which the fish can meet requirements for both acceleration and stabilization in limited space. Quantitative 3D PIV wake measurements reveal how variations in tail kinematics relate to thrust production throughout the course of a jumping maneuver and over a range of jump heights. By performing measurements in 3D, the timing, interactions, and relative contributions to thrust and lateral forces from each fin can be evaluated, elucidating the complex hydrodynamics that enable archer fish water exit.

  18. Effect of Maize Hybrid Maturity and Grain Hardness on Fumonisin and Zearalenone Contamination

    Directory of Open Access Journals (Sweden)

    Amedeo Reyneri

    2011-02-01

    Full Text Available The level of resistance in commercial hybrids for Fusarium ear rot is still not in general adequate to prevent unacceptable toxin concentrations in field. The purpose of this experiment was to verify the behaviour of commercial dent maize hybrids for fumonisin and zearalenone contamination and to identify the variety traits that influence the production of these toxins. Field experiments were carried out in 2000, 2001 and 2002 to evaluate the effect of maize hybrid maturity and endosperm hardness on European Corn Borer (ECB incidence, fungal ear rot incidence and severity and on fumonisin B1 and zearalenone contents. Nineteen yellow soft commercial hybrids, from the 500, 600 and 700 FAO maturity groups, were compared in 4 sites in NW Italy. Hybrid were grouped in 3 endosperm hardness categories (hard, intermediate, soft in function of Hard/Soft (H/S endosperm ratio. No effect due to endosperm hardness or hybrid maturity on the ECB infestation or fungal ear rot incidence and severity was observed. Grain hardness significant influenced fumonisin B1 content: hard endosperm hybrids showed 50% lower contamination than soft hybrids. The presence of fumonisin B1 in the grain of different maturity hybrids only resulted to be significantly different in 2001 experiment, with a mean concentration 2 times higher in the later hybrids (FAO rating 700 compared to the medium and medium-late hybrids. The zearalenone content never resulted to be significantly different in function of the endosperm hardness, while, late maturing hybrids, in which grain moisture content decreases slowly below 30%, are more susceptible to zearalenone contamination. This research has highlighted the presence of variety traits that can influence mycotoxin contamination. An accurate choice of hybrid, considering the territorial and cultivation context, could contribute to achieve products, that contain mycotoxins, which do not exceed the maximum international and UE regulation levels.

  19. Case for the fusion hybrid

    International Nuclear Information System (INIS)

    Rose, R.P.

    1981-01-01

    The use of nuclear fusion to produce fuel for nuclear fission power stations is discussed in the context of a crucial need for future energy options. The fusion hybrid is first considered as an element in the future of nuclear fission power to provide long term assurance of adequate fuel supplies for both breeder and convertor reactors. Generic differences in neutronic characteristics lead to a fuel production potential of fusion-fission hybrid systems which is significantly greater than that obtainable with fission systems alone. Furthermore, cost benefit studies show a variety of scenarios in which the hybrid offers sufficient potential to justify development costs ranging in the tens of billions of dollars. The hybrid is then considered as an element in the ultimate development of fusion electric power. The hybrid offers a near term application of fusion where experience with the requisite technologies can be derived as a vital step in mapping a credible route to eventual commercial feasibility of pure fusion systems. Finally, the criteria for assessment of future energy options are discussed with prime emphasis on the need for rational comparision of alternatives

  20. Small nonnative fishes as predators of larval razorback suckers

    Science.gov (United States)

    Carpenter, J.; Mueller, G.A.

    2008-01-01

    The razorback sucker (Xyrauchen texanus), an endangered big-river fish of the Colorado River basin, has demonstrated no sustainable recruitment in 4 decades, despite presence of spawning adults and larvae. Lack of adequate recruitment has been attributed to several factors, including predation by nonnative fishes. Substantial funding and effort has been expended on mechanically removing nonnative game fishes, typically targeting large predators. As a result, abundance of larger predators has declined, but the abundance of small nonnative fishes has increased in some areas. We conducted laboratory experiments to determine if small nonnative fishes would consume larval razorback suckers. We tested adults of three small species (threadfin shad, Dorosoma petenense; red shiner, Cyprinella lutrensis; fathead minnow, Pimephales promelas) and juveniles of six larger species (common carp, Cyprinus carpio; yellow bullhead, Ameiurus natalis; channel catfish, Ictalurus punctatus; rainbow trout, Oncorhynchus mykiss; green sunfish, Lepomis cyanellus; bluegill, L. macrochirus). These nonnative fishes span a broad ecological range and are abundant within the historical range of the razorback sucker. All nine species fed on larval razorback suckers (total length, 9-16 mm). Our results suggest that predation by small nonnative fishes could be responsible for limiting recovery of this endangered species.

  1. Immunity, feed, and husbandry in fish health management of cultured Epinephelus fuscoguttatus with reference to Epinephelus coioides

    Directory of Open Access Journals (Sweden)

    Catherine Cheng Yun Chieng

    2018-03-01

    Full Text Available Groupers are dispersed worldwide in the tropical and subtropical waters. They are prized in the live reef fish trade, making them candidates with high market value and consumer demand. In the Asian-Pacific region, the brown-marbled grouper (Epinephelus fuscoguttatus is widely adapted as an aquaculture species. However, health management remains a major concern in the stressful intensive culture process. The present review discusses techniques and current knowledge in the cultivation of E. fuscoguttatus towards healthy fish growth focusing on aspects of immunity, feed, and husbandry. Understanding how the fish immune system responds during infections provides insights into the intricate ways fish resist pathogens. This information is helpful when developing vaccination strategies or immunostimulant compounds to strengthen fish immunity. Feeds that are formulated according to the needs of the fish ensure optimal growth and using suitable alternative ingredients may lower production cost without compromising fish health. Good husbandry practices contribute to a favourable environment for the fish to grow, while interspecific hybridization may be a convenient approach to generate hardier species. Nevertheless, it has been noted that research in E. fuscoguttatus is relatively scarce in contrast to the closely-related orange-spotted grouper (E. coioides, which is used in the present article as a reference. Ultimately, the identified gaps in knowledge between the two species warrant species-specific research in E. fuscoguttatus to promote fish health and ensure continued success in aquaculture. Keywords: Epinephelus fuscoguttatus, Epinephelus coioides, Health, Immunity, Feed, Husbandry

  2. Rapid detection of chromosome rearrangement in medical diagnostic X-ray workers by using fluorescence in situ hybridization and study on dose estimation

    International Nuclear Information System (INIS)

    Wang Zhiquan; Sun Yuanming; Li Jin

    1998-01-01

    Objective: Biological doses were estimated for medical diagnostic X-ray workers. Methods: Chromosome rearrangements in X-ray workers were analysed by fluorescence in situ hybridization (FISH) with composite whole chromosome paintings number 4 and number 7. Results: The frequency of translocation in medical diagnostic X-ray workers was much higher than that in control group (P<0.01). The biological doses to individual X-ray workers were calculated by their translocation frequency. The translocation frequencies of both FISH and G-banding were in good agreement. Conclusion: The biological doses to X-ray workers are estimated by FISH first when their dosimetry records are not documented

  3. Fishing degrades size structure of coral reef fish communities.

    Science.gov (United States)

    Robinson, James P W; Williams, Ivor D; Edwards, Andrew M; McPherson, Jana; Yeager, Lauren; Vigliola, Laurent; Brainard, Russell E; Baum, Julia K

    2017-03-01

    Fishing pressure on coral reef ecosystems has been frequently linked to reductions of large fishes and reef fish biomass. Associated impacts on overall community structure are, however, less clear. In size-structured aquatic ecosystems, fishing impacts are commonly quantified using size spectra, which describe the distribution of individual body sizes within a community. We examined the size spectra and biomass of coral reef fish communities at 38 US-affiliated Pacific islands that ranged in human presence from near pristine to human population centers. Size spectra 'steepened' steadily with increasing human population and proximity to market due to a reduction in the relative biomass of large fishes and an increase in the dominance of small fishes. Reef fish biomass was substantially lower on inhabited islands than uninhabited ones, even at inhabited islands with the lowest levels of human presence. We found that on populated islands size spectra exponents decreased (analogous to size spectra steepening) linearly with declining biomass, whereas on uninhabited islands there was no relationship. Size spectra were steeper in regions of low sea surface temperature but were insensitive to variation in other environmental and geomorphic covariates. In contrast, reef fish biomass was highly sensitive to oceanographic conditions, being influenced by both oceanic productivity and sea surface temperature. Our results suggest that community size structure may be a more robust indicator than fish biomass to increasing human presence and that size spectra are reliable indicators of exploitation impacts across regions of different fish community compositions, environmental drivers, and fisheries types. Size-based approaches that link directly to functional properties of fish communities, and are relatively insensitive to abiotic variation across biogeographic regions, offer great potential for developing our understanding of fishing impacts in coral reef ecosystems. © 2016

  4. Fish. Animal Life in Action[TM]. Schlessinger Science Library. [Videotape].

    Science.gov (United States)

    2000

    This 23-minute videotape for grades 5-8, presents the myriad of animal life that exists on the planet. Students can view and perform experiments and investigations that help explain animal traits and habits. There are many types of fish that live in oceans, lakes, and streams. Students learn about fish characteristics and environmental habitats,…

  5. Essential Fish Habitat (EFH) Areas Protected From Fishing

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Designated Essential Fish Habitat (EFH) areas where fishing or the use of fishing gears has been restricted or modified in order to minimize the adverse effects of...

  6. Chromogenic in situ hybridization is a reliable assay for detection of ALK rearrangements in adenocarcinomas of the lung.

    Science.gov (United States)

    Schildhaus, Hans-Ulrich; Deml, Karl-Friedrich; Schmitz, Katja; Meiboom, Maren; Binot, Elke; Hauke, Sven; Merkelbach-Bruse, Sabine; Büttner, Reinhard

    2013-11-01

    Reliable detection of anaplastic lymphoma kinase (ALK) rearrangements is a prerequisite for personalized treatment of lung cancer patients, as ALK rearrangements represent a predictive biomarker for the therapy with specific tyrosine kinase inhibitors. Currently, fluorescent in situ hybridization (FISH) is considered to be the standard method for assessing formalin-fixed and paraffin-embedded tissue for ALK inversions and translocations. However, FISH requires a specialized equipment, the signals fade rapidly and it is difficult to detect overall morphology and tumor heterogeneity. Chromogenic in situ hybridization (CISH) has been successfully introduced as an alternative test for the detection of several genetic aberrations. This study validates a newly developed ALK CISH assay by comparing FISH and CISH signal patterns in lung cancer samples with and without ALK rearrangements. One hundred adenocarcinomas of the lung were included in this study, among them 17 with known ALK rearrangement. FISH and CISH were carried out and evaluated according to the manufacturers' recommendations. For both assays, tumors were considered positive if ≥15% of tumor cells showed either isolated 3' signals or break-apart patterns or a combination of both. A subset of tumors was exemplarily examined by using a novel EML4 (echinoderm microtubule-associated protein-like 4) CISH probe. Red, green and fusion CISH signals were clearcut and different signal patterns were easily recognized. The percentage of aberrant tumor cells was statistically highly correlated (PCISH. On the basis of 86 samples that were evaluable by ALK CISH, we found a 100% sensitivity and 100% specificity of this assay. Furthermore, EML4 rearrangements could be recognized by CISH. CISH is a highly reliable, sensitive and specific method for the detection of ALK gene rearrangements in pulmonary adenocarcinomas. Our results suggest that CISH might serve as a suitable alternative to FISH, which is the current gold

  7. Hybrid quantum systems of ions and atoms

    OpenAIRE

    Sias, Carlo; Köhl, Michael

    2014-01-01

    In this chapter we review the progress in experiments with hybrid systems of trapped ions and ultracold neutral atoms. We give a theoretical overview over the atom-ion interactions in the cold regime and give a summary of the most important experimental results. We conclude with an overview of remaining open challenges and possible applications in hybrid quantum systems of ions and neutral atoms.

  8. FISHing for gutta-percha-adhered biofilms in purulent post-treatment apical periodontitis.

    Science.gov (United States)

    Zehnder, M; Rechenberg, D-K; Thurnheer, T; Lüthi-Schaller, H; Belibasakis, G N

    2017-06-01

    This study investigated the possibility of depicting individual taxa in clinically relevant biofilms using fluorescent in situ hybridization (FISH). Gutta-percha samples were collected from the apical aspect of root canals associated with a chronic apical abscess (test samples, n = 8). Corresponding control samples were obtained from previously filled root canals with apparently normal periapical tissues (n = 3). The transport medium was investigated for detached biofilm fragments using FISH staining and conventional epifluorescence microscopy. Gutta-percha samples were stained by multiplex FISH, and inspected using confocal laser scanning microscopy. FISH of the transport medium confirmed the presence of the main species formerly identified by conventional methods in post-treatment purulent endodontic infections, most prominently Fusobacterium spp., Bacteroidetes and Prevotellaceae. Treponemes were identified in five of eight cases associated with purulent infections, but Enterococcus faecalis and Staphylococcus spp. were not identified. The biofilms on gutta-percha from root canals associated with apical periodontitis showed dense aggregates of variable composition. Control samples contained few, if any, bacteria in the transport medium, and featured no biofilms on the respective gutta-percha specimens. The current study revealed some direct, visual in situ information on the nature of biofilms associated with purulent periapical infections in man. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  9. Hybrid recreation by reverse breeding in Arabidopsis thaliana.

    Science.gov (United States)

    Wijnker, Erik; Deurhof, Laurens; van de Belt, Jose; de Snoo, C Bastiaan; Blankestijn, Hetty; Becker, Frank; Ravi, Maruthachalam; Chan, Simon W L; van Dun, Kees; Lelivelt, Cilia L C; de Jong, Hans; Dirks, Rob; Keurentjes, Joost J B

    2014-04-01

    Hybrid crop varieties are traditionally produced by selecting and crossing parental lines to evaluate hybrid performance. Reverse breeding allows doing the opposite: selecting uncharacterized heterozygotes and generating parental lines from them. With these, the selected heterozygotes can be recreated as F1 hybrids, greatly increasing the number of hybrids that can be screened in breeding programs. Key to reverse breeding is the suppression of meiotic crossovers in a hybrid plant to ensure the transmission of nonrecombinant chromosomes to haploid gametes. These gametes are subsequently regenerated as doubled-haploid (DH) offspring. Each DH carries combinations of its parental chromosomes, and complementing pairs can be crossed to reconstitute the initial hybrid. Achiasmatic meiosis and haploid generation result in uncommon phenotypes among offspring owing to chromosome number variation. We describe how these features can be dealt with during a reverse-breeding experiment, which can be completed in six generations (∼1 year).

  10. Hybrid Simulation of Composite Structures

    DEFF Research Database (Denmark)

    Høgh, Jacob Herold

    experiment. The technique has primarily been used within earthquake engineering but many other fields of engineering have utilized the method with benefit. However, these previous efforts have focused on structures with a simple boundary between the numerical and physical substructure i.e. few degrees...... the transfer system and the control and monitoring techniques in the shared boundary is therefore a key issue in this type of hybrid simulation. During the research, hybrid simulation platforms have been programmed capable of running on different time scales with advanced control and monitoring techniques...

  11. Flow Behavior Around a Fast-Starting Robotic Fish

    Science.gov (United States)

    Ma, Ganzhong; Currier, Todd; Modarres-Sadeghi, Yahya

    2017-11-01

    A robotic fish is used to study the flow behavior around the body of a fast-starting fish as it experiences a fast-start. The robotic fish is designed and built emulating a Northern Pike, Esox Lucius, which can accelerate at up to 245 m/s2. In previous studies, we had focused on the flow around the tail during the fast-start, by using a tail which acted flexibly in the preparatory stage and rigidly in the propulsive stage. We have extended that study by including the fish body in the experimental setup, where the body can bend into a C-shape, so that the influence of the body motion on the resulting flow around the structure can be understood as well. In the tests, the fish can rotate about a vertical axis, where a multi-axis force sensor measures flow forces acting on the body. Synchronized with the force measurement, flow visualizations using bubble image velocimetry are conducted, and the observed shed vortices are related to the peak forces observed during the maneuver.

  12. Allele-specific expression at the androgen receptor alpha gene in a hybrid unisexual fish, the Amazon molly (Poecilia formosa.

    Directory of Open Access Journals (Sweden)

    Fangjun Zhu

    Full Text Available The all-female Amazon molly (Poecilia formosa is the result of a hybridization of the Atlantic molly (P. mexicana and the sailfin molly (P. latipinna approximately 120,000 years ago. As a gynogenetic species, P. formosa needs to copulate with heterospecific males including males from one of its bisexual ancestral species. However, the sperm only triggers embryogenesis of the diploid eggs. The genetic information of the sperm donor typically will not contribute to the next generation of P. formosa. Hence, P. formosa possesses generally one allele from each of its ancestral species at any genetic locus. This raises the question whether both ancestral alleles are equally expressed in P. formosa. Allele-specific expression (ASE has been previously assessed in various organisms, e.g., human and fish, and ASE was found to be important in the context of phenotypic variability and disease. In this study, we utilized Real-Time PCR techniques to estimate ASE of the androgen receptor alpha (arα gene in several distinct tissues of Amazon mollies. We found an allelic bias favoring the maternal ancestor (P. mexicana allele in ovarian tissue. This allelic bias was not observed in the gill or the brain tissue. Sequencing of the promoter regions of both alleles revealed an association between an Indel in a known CpG island and differential expression. Future studies may reveal whether our observed cis-regulatory divergence is caused by an ovary-specific trans-regulatory element, preferentially activating the allele of the maternal ancestor.

  13. Long Term Resource Monitoring Program procedures: fish monitoring

    Science.gov (United States)

    Ratcliff, Eric N.; Glittinger, Eric J.; O'Hara, T. Matt; Ickes, Brian S.

    2014-01-01

    This manual constitutes the second revision of the U.S. Army Corps of Engineers’ Upper Mississippi River Restoration-Environmental Management Program (UMRR-EMP) Long Term Resource Monitoring Program (LTRMP) element Fish Procedures Manual. The original (1988) manual merged and expanded on ideas and recommendations related to Upper Mississippi River fish sampling presented in several early documents. The first revision to the manual was made in 1995 reflecting important protocol changes, such as the adoption of a stratified random sampling design. The 1995 procedures manual has been an important document through the years and has been cited in many reports and scientific manuscripts. The resulting data collected by the LTRMP fish component represent the largest dataset on fish within the Upper Mississippi River System (UMRS) with more than 44,000 collections of approximately 5.7 million fish. The goal of this revision of the procedures manual is to document changes in LTRMP fish sampling procedures since 1995. Refinements to sampling methods become necessary as monitoring programs mature. Possible refinements are identified through field experiences (e.g., sampling techniques and safety protocols), data analysis (e.g., planned and studied gear efficiencies and reallocations of effort), and technological advances (e.g., electronic data entry). Other changes may be required because of financial necessity (i.e., unplanned effort reductions). This version of the LTRMP fish monitoring manual describes the most current (2014) procedures of the LTRMP fish component.

  14. Assessment on Hybrid E-Learning Instrument

    OpenAIRE

    Intan Farahana Kamsin; Rosseni Din

    2015-01-01

    This study aims to improve Hybrid e-Learning 9.3. A total of 233 students of International Islamic University Malaysia, Gombak who have the experience in hybrid teaching and learning were involved as respondents. Rasch Measurement Model was used for this study. Validity analyses conducted were on (i) the compatibility of the items, (ii) mapping of items and respondents, (iii) scaling of instruments, and (iv) unidimentional items. The findings of the study show that (i) the items developed cor...

  15. Six-Degree-of-Freedom Sensor Fish Design: Governing Equations and Motion Modeling

    Energy Technology Data Exchange (ETDEWEB)

    Deng, Zhiqun; Richmond, Marshall C.; Simmons, Carver S.; Carlson, Thomas J.

    2004-08-19

    The Sensor Fish device is being used at Northwest hydropower projects to better understand the conditions fish experience during passage through hydroturbines and other dam bypass alternatives. Since its initial development in 1997, the Sensor Fish has undergone numerous design changes to improve its function and extend the range of its use. The most recent Sensor Fish design, the three degree of freedom (3DOF) device, has been used successfully to characterize the environment fish experience when passing through turbines, in spill, or in engineered fish bypass facilities at dams. Pacific Northwest National Laboratory (PNNL) is in the process of redesigning the current 3DOF Sensor Fish device package to improve its field performance. Rate gyros will be added to the new six degree of freedom (6DOF) device so that it will be possible to observe the six linear and angular accelerations of the Sensor Fish as it passes the dam. Before the 6DOF Sensor Fish device can be developed and deployed, governing equations of motion must be developed in order to understand the design implications of instrument selection and placement within the body of the device. In this report, we describe a fairly general formulation for the coordinate systems, equations of motion, force and moment relationships necessary to simulate the 6DOF movement of an underwater body. Some simplifications are made by considering the Sensor Fish device to be a rigid, axisymmetric body. The equations of motion are written in the body-fixed frame of reference. Transformations between the body-fixed and interial reference frames are performed using a formulation based on quaternions. Force and moment relationships specific to the Sensor Fish body are currently not available. However, examples of the trajectory simulations using the 6DOF equations are presented using existing low and high-Reynolds number force and moment correlations. Animation files for the test cases are provided in an attached CD. The next

  16. Robotic Fish to Aid Animal Behavior Studies and Informal Science Learning

    Science.gov (United States)

    Phamduy, Paul

    The application of robotic fish in the fields of animal behavior and informal science learning are new and relatively untapped. In the context of animal behavior studies, robotic fish offers a consistent and customizable stimulus that could contribute to dissect the determinants of social behavior. In the realm of informal science learning, robotic fish are gaining momentum for the possibility of educating the general public simultaneously on fish physiology and underwater robotics. In this dissertation, the design and development of a number of robotic fish platforms and prototypes and their application in animal behavioral studies and informal science learning settings are presented. Robotic platforms for animal behavioral studies focused on the utilization replica or same scale prototypes. A novel robotic fish platform, featuring a three-dimensional swimming multi-linked robotic fish, was developed with three control modes varying in the level of robot autonomy offered. This platform was deployed at numerous science festivals and science centers, to obtain data on visitor engagement and experience.

  17. Uptake of {sup 137}Cs by fresh water fish

    Energy Technology Data Exchange (ETDEWEB)

    Man, C.K.; Kwok, Y.H

    2000-02-01

    The uptake and discharge rates of {sup 137}Cs by fresh water fish at different radionuclide concentrations have been studied. A dual compartment model was used to fit the experimental data. The discharge rates have been found to be negligible for the duration of the experiment of 10 days. The uptake rates were independent of radionuclide concentrations for a particular type of fresh water fish and were different for different types of fish. The uptake rates of carp, tilapia and snakehead were 1.58, 1.66 and 2.23, in unit of 10{sup -6} h{sup -1}, respectively. It was also estimated that the consumption of fresh water fish, even if the water were contaminated as much as that in the Chernobyl accident, leads to negligible latent cancer fatality to the Hong Kong population.

  18. Molecular and morphological approaches for species delimitation and hybridization investigations of two Cichla species

    Directory of Open Access Journals (Sweden)

    Andrea A. F. Mourão

    Full Text Available ABSTRACT The hybridization is a widely-discussed issue in several studies with fish species. For some authors, hybridization may be related with diversification and speciation of several groups, or also with the extinction of populations or species. Difficulties to differentiate species and hybrids may be a problem to correctly apply a management of wild species, because hybrid lineages, especially the advanced ones, may resemble the parental species. The genus Cichla Bloch & Schneider, 1801 constitutes an interesting experimental model, considering that hybridization and taxonomic uncertainties hinder a correct identification. Considering these problems, in this study, we developed genetic methodologies and applied meristic and morphometric approaches in wild samples in order to identify species and for test a possible hybridization between Cichla kelberi Kullander & Ferreira, 2006 and Cichla piquiti Kullander & Ferreira, 2006. For this, C. kelberi, C. piquiti and potential hybrid ( carijó individuals were collected in Paraná and Tietê rivers (SP, Brazil. For meristic and morphometric methods, the individuals were analyzed using the statistical software Pcord 5:31, while for molecular methods, primers for PCR-multiplex were designed and enzyme for PCR-RFLP were selected, under the species-specific nucleotide. All results indicated that the carijó is not an interspecific hybrid, because it presented identical genetic pattern and morphology closed to C. piquiti. Thus, we propose that carijó is a C. piquiti morphotype. In addition, this study promotes a new molecular tool that could be used in future research, monitoring and management programs of the genus Cichla.

  19. Dual color chromogenic in situ hybridization for determination of HER2 status in breast cancer: a large comparative study to current state of the art fluorescence in situ hybridization

    Directory of Open Access Journals (Sweden)

    Mollerup Jens

    2012-02-01

    Full Text Available Abstract Background Chromogenic in situ hybridization (CISH is fast becoming a well established technique for easy and sensitive determination of HER2 gene status in breast cancer. However, for the chromogenic method to achieve status as a safe and reliable technique, the method needs to be validated against already known and validated FISH techniques. Methods Here it is reported from a comparative study where HER2 gene status obtained by HER2 CISH pharmDx™ Kit was compared to HER2 gene status obtained by the FDA approved HER2 FISH pharmDx™ Kit and the PathVysion HER-2 DNA probe Kit. The study included 365 formalin fixed and paraffin-embedded invasive breast cancer tissue specimens collected consecutively at a US reference laboratory. Results The data obtained revealed an overall HER2 status concordance of approximately 98% for comparisons of HER2 CISH pharmDx™ Kit to both HER2 FISH pharmDx™ Kit and PathVysion HER-2 DNA Probe Kit. Conclusions The concordance between results obtained using the recently FDA approved HER2 CISH pharmDx™ Kit with previously FDA approved FISH techniques for HER2 gene status determination indicate that the HER2 CISH pharmDx™ Kit is a reliable chromogenic alternative to fluorescence-based methods.

  20. Dual color chromogenic in situ hybridization for determination of HER2 status in breast cancer: a large comparative study to current state of the art fluorescence in situ hybridization

    Science.gov (United States)

    2012-01-01

    Background Chromogenic in situ hybridization (CISH) is fast becoming a well established technique for easy and sensitive determination of HER2 gene status in breast cancer. However, for the chromogenic method to achieve status as a safe and reliable technique, the method needs to be validated against already known and validated FISH techniques. Methods Here it is reported from a comparative study where HER2 gene status obtained by HER2 CISH pharmDx™ Kit was compared to HER2 gene status obtained by the FDA approved HER2 FISH pharmDx™ Kit and the PathVysion HER-2 DNA probe Kit. The study included 365 formalin fixed and paraffin-embedded invasive breast cancer tissue specimens collected consecutively at a US reference laboratory. Results The data obtained revealed an overall HER2 status concordance of approximately 98% for comparisons of HER2 CISH pharmDx™ Kit to both HER2 FISH pharmDx™ Kit and PathVysion HER-2 DNA Probe Kit. Conclusions The concordance between results obtained using the recently FDA approved HER2 CISH pharmDx™ Kit with previously FDA approved FISH techniques for HER2 gene status determination indicate that the HER2 CISH pharmDx™ Kit is a reliable chromogenic alternative to fluorescence-based methods. PMID:22333181