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Sample records for harboring luciferase reporter

  1. Transgenic Mouse Model Harboring the Transcriptional Fusion Ccl20-Luciferase as a Novel Reporter of Pro-Inflammatory Response

    Science.gov (United States)

    Crispo, Martina; Van Maele, Laurye; Tabareau, Julien; Cayet, Delphine; Errea, Agustina; Ferreira, Ana María; Rumbo, Martin; Sirard, Jean Claude

    2013-01-01

    The chemokine CCL20, the unique ligand of CCR6 functions as an attractant of immune cells. Expression of CCL20 is induced by Toll-like Receptor (TLR) signaling or proinflammatory cytokine stimulation. However CCL20 is also constitutively produced at specific epithelial sites of mucosa. This expression profile is achieved by transcriptional regulation. In the present work we characterized regulatory features of mouse Ccl20 gene. Transcriptional fusions between the mouse Ccl20 promoter and the firefly luciferase (luc) encoding gene were constructed and assessed in in vitro and in vivo assays. We found that liver CCL20 expression and luciferase activity were upregulated by systemic administration of the TLR5 agonist flagellin. Using shRNA and dominant negative form specific for mouse TLR5, we showed that this expression was controlled by TLR5. To address in situ the regulation of gene activity, a transgenic mouse line harboring a functional Ccl20-luc fusion was generated. The luciferase expression was highly concordant with Ccl20 expression in different tissues. Our data indicate that the transgenic mouse model can be used to monitor activation of innate response in vivo. PMID:24265691

  2. In vivo bioimaging with tissue-specific transcription factor activated luciferase reporters.

    OpenAIRE

    Buckley, SM; Delhove, JM; Perocheau, DP; Karda, R; Rahim, AA; Howe, SJ; Ward, NJ; Birrell, MA; Belvisi, MG; Arbuthnot, P; Johnson, MR; Waddington, SN; McKay, TR

    2015-01-01

    The application of transcription factor activated luciferase reporter cassettes in vitro is widespread but potential for in vivo application has not yet been realized. Bioluminescence imaging enables non-invasive tracking of gene expression in transfected tissues of living rodents. However the mature immune response limits luciferase expression when delivered in adulthood. We present a novel approach of tissue-targeted delivery of transcription factor activated luciferase reporter lentiviruse...

  3. High-Throughput Screening of a Luciferase Reporter of Gene Silencing on the Inactive X Chromosome.

    Science.gov (United States)

    Keegan, Alissa; Plath, Kathrin; Damoiseaux, Robert

    2018-01-01

    Assays of luciferase gene activity are a sensitive and quantitative reporter system suited to high-throughput screening. We adapted a luciferase assay to a screening strategy for identifying factors that reactivate epigenetically silenced genes. This epigenetic luciferase reporter is subject to endogenous gene silencing mechanisms on the inactive X chromosome (Xi) in primary mouse cells and thus captures the multilayered nature of chromatin silencing in development. Here, we describe the optimization of an Xi-linked luciferase reactivation assay in 384-well format and adaptation of the assay for high-throughput siRNA and chemical screening. Xi-luciferase reactivation screening has applications in stem cell biology and cancer therapy. We have used the approach described here to identify chromatin-modifying proteins and to identify drug combinations that enhance the gene reactivation activity of the DNA demethylating drug 5-aza-2'-deoxycytidine.

  4. Two panels of steroid receptor luciferase reporter cell lines for compound profiling

    Czech Academy of Sciences Publication Activity Database

    Sedlák, David; Paguio, A.; Bartůněk, Petr

    2011-01-01

    Roč. 14, č. 2 (2011), s. 248-266 ISSN 1386-2073 R&D Projects: GA MŠk(CZ) LC06077 Institutional research plan: CEZ:AV0Z50520514 Keywords : nuclear hormone receptor * steroid receptor * cell-based luciferase reporter assay Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.785, year: 2011

  5. A Dual Luciferase Reporter System for B. burgdorferi Measures Transcriptional Activity during Tick-Pathogen Interactions

    Directory of Open Access Journals (Sweden)

    Philip P. Adams

    2017-05-01

    Full Text Available Knowledge of the transcriptional responses of vector-borne pathogens at the vector-pathogen interface is critical for understanding disease transmission. Borrelia (Borreliella burgdorferi, the causative agent of Lyme disease in the United States, is transmitted by the bite of infected Ixodes sp. ticks. It is known that B. burgdorferi has altered patterns of gene expression during tick acquisition, persistence and transmission. Recently, we and others have discovered in vitro expression of RNAs found internal, overlapping, and antisense to annotated open reading frames in the B. burgdorferi genome. However, there is a lack of molecular genetic tools for B. burgdorferi for quantitative, strand-specific, comparative analysis of these transcripts in distinct environments such as the arthropod vector. To address this need, we have developed a dual luciferase reporter system to quantify B. burgdorferi promoter activities in a strand-specific manner. We demonstrate that constitutive expression of a B. burgdorferi codon-optimized Renilla reniformis luciferase gene (rlucBb allows normalization of the activity of a promoter of interest when fused to the B. burgdorferi codon-optimized Photinus pyralis luciferase gene (flucBb on the same plasmid. Using the well characterized, differentially regulated, promoters for flagellin (flaBp, outer surface protein A (ospAp and outer surface protein C (ospCp, we document the efficacy of the dual luciferase system for quantitation of promoter activities during in vitro growth and in infected ticks. Cumulatively, the dual luciferase method outlined herein is the first dual reporter system for B. burgdorferi, providing a novel and highly versatile approach for strand-specific molecular genetic analyses.

  6. Relative quantification of protein-protein interactions using a dual luciferase reporter pull-down assay system.

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    Shuaizheng Jia

    Full Text Available The identification and quantitative analysis of protein-protein interactions are essential to the functional characterization of proteins in the post-proteomics era. The methods currently available are generally time-consuming, technically complicated, insensitive and/or semi-quantitative. The lack of simple, sensitive approaches to precisely quantify protein-protein interactions still prevents our understanding of the functions of many proteins. Here, we develop a novel dual luciferase reporter pull-down assay by combining a biotinylated Firefly luciferase pull-down assay with a dual luciferase reporter assay. The biotinylated Firefly luciferase-tagged protein enables rapid and efficient isolation of a putative Renilla luciferase-tagged binding protein from a relatively small amount of sample. Both of these proteins can be quantitatively detected using the dual luciferase reporter assay system. Protein-protein interactions, including Fos-Jun located in the nucleus; MAVS-TRAF3 in cytoplasm; inducible IRF3 dimerization; viral protein-regulated interactions, such as MAVS-MAVS and MAVS-TRAF3; IRF3 dimerization; and protein interaction domain mapping, are studied using this novel assay system. Herein, we demonstrate that this dual luciferase reporter pull-down assay enables the quantification of the relative amounts of interacting proteins that bind to streptavidin-coupled beads for protein purification. This study provides a simple, rapid, sensitive, and efficient approach to identify and quantify relative protein-protein interactions. Importantly, the dual luciferase reporter pull-down method will facilitate the functional determination of proteins.

  7. Luciferase NanoLuc as a reporter for gene expression and protein levels in Saccharomyces cerevisiae.

    Science.gov (United States)

    Masser, Anna E; Kandasamy, Ganapathi; Kaimal, Jayasankar Mohanakrishnan; Andréasson, Claes

    2016-05-01

    Reporter proteins are essential tools in the study of biological processes and are employed to monitor changes in gene expression and protein levels. Luciferases are reporter proteins that enable rapid and highly sensitive detection with an outstanding dynamic range. Here we evaluated the usefulness of the 19 kDa luciferase NanoLuc (Nluc), derived from the deep sea shrimp Oplophorus gracilirostris, as a reporter protein in yeast. Cassettes with codon-optimized genes expressing yeast Nluc (yNluc) or its destabilized derivative yNlucPEST have been assembled in the context of the dominant drug resistance marker kanMX. The reporter proteins do not impair the growth of yeast cells and exhibit half-lives of 40 and 5 min, respectively. The commercial substrate Nano-Glo® is compatible with detection of yNluc bioluminescence in yeast using standard commercial substrate. © 2016 The Authors. Yeast published by John Wiley & Sons Ltd. © 2016 The Authors. Yeast published by John Wiley & Sons Ltd.

  8. Flexible Measurement of Bioluminescent Reporters Using an Automated Longitudinal Luciferase Imaging Gas- and Temperature-optimized Recorder (ALLIGATOR).

    Science.gov (United States)

    Crosby, Priya; Hoyle, Nathaniel P; O'Neill, John S

    2017-12-13

    Luciferase-based reporters of cellular gene expression are in widespread use for both longitudinal and end-point assays of biological activity. In circadian rhythms research, for example, clock gene fusions with firefly luciferase give rise to robust rhythms in cellular bioluminescence that persist over many days. Technical limitations associated with photomultiplier tubes (PMT) or conventional microscopy-based methods for bioluminescence quantification have typically demanded that cells and tissues be maintained under quite non-physiological conditions during recording, with a trade-off between sensitivity and throughput. Here, we report a refinement of prior methods that allows long-term bioluminescence imaging with high sensitivity and throughput which supports a broad range of culture conditions, including variable gas and humidity control, and that accepts many different tissue culture plates and dishes. This automated longitudinal luciferase imaging gas- and temperature-optimized recorder (ALLIGATOR) also allows the observation of spatial variations in luciferase expression across a cell monolayer or tissue, which cannot readily be observed by traditional methods. We highlight how the ALLIGATOR provides vastly increased flexibility for the detection of luciferase activity when compared with existing methods.

  9. A novel luciferase knock-in reporter system for studying transcriptional regulation of the human Sox2 gene.

    Science.gov (United States)

    Xiao, Dan; Zhang, Weifeng; Li, Yan; Liu, Kuan; Zhao, Junli; Sun, Xiaohong; Shan, Linlin; Mao, Qinwen; Xia, Haibin

    2016-02-10

    Sox2 is an important transcriptional factor that has multiple functions in stem cell maintenance and tumorigenesis. To investigate the transcriptional regulation of the Sox2 gene, a luciferase knock-in reporter system was established in HEK293 cells by placing the luciferase gene in the genome under the control of the Sox2 gene promoter using a transcription activator-like effector nuclease (TALEN)-mediated genome editing technique. PCR and Southern blot results confirmed the site-specific integration of a single copy of the exogenous luciferase gene into the genome. To prove the reliability and sensitivity of this novel luciferase knock-in system, a CRISPR/Cas transcription activation system for the Sox2 gene was constructed and applied to the knock-in system. The results indicated that luciferase activity was directly correlated with the activity of the Sox2 endogenous promoter. This novel system will be a useful tool to study the transcriptional regulation of Sox2, and has great potential in medical and industrial applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. In Vivo Imaging of Retinoic Acid Receptor Activity using a Sodium/Iodide Symporter and Luciferase Dual Imaging Reporter Gene

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    Min Kyung So

    2004-07-01

    Full Text Available Retinoic acids are natural derivatives of vitamin A, and play important roles in modulating tumor cell growth by regulating differentiation, thus suggesting the potential use of these derivatives in cancer therapy and prevention. To visualize the intranuclear responses of functional retinoic acid receptors, we have developed a dual-imaging reporter gene system based on the use of sodium/iodide symporter (NIS and luciferase in cancer cell lines. NIS and luciferase genes were linked with an internal ribosome entry site, and placed under the control of an artificial cis-acting retinoic acid responsive element (pRARE/NL. After retinoic acid treatment, I-125 uptake by pRARE/NL transfected cells was found to have increased by up to about five times that of nontreated cells. The bioluminescence intensity of pRARE/NL transfected cells showed dose-dependency. In vivo luciferase images showed higher intensity in retinoic acid treated SK-RARE/NL tumors, and scintigraphic images of SK-RARE/NL tumors showed increased Tc-99m uptake after retinoic acid treatment. The NIS/luciferase imaging reporter system was sufficiently sensitive to allow the visualization of intranuclear retinoic acid receptor activity. This cis-enhancer imaging reporter system may be useful in vitro and in vivo for the evaluation of retinoic acid responses in such areas as cellular differentiation and chemoprevention.

  11. Development of HSPA1A promoter-driven luciferase reporter gene assays in human cells for assessing the oxidative damage induced by silver nanoparticles

    International Nuclear Information System (INIS)

    Xin, Lili; Wang, Jianshu; Zhang, Leshuai W.; Che, Bizhong; Dong, Guangzhu; Fan, Guoqiang; Cheng, Kaiming

    2016-01-01

    The exponential increase in the total number of engineered nanoparticles in consumer products requires novel tools for rapid and cost-effective toxicology screening. In order to assess the oxidative damage induced by nanoparticles, toxicity test systems based on a human HSPA1A promoter-driven luciferase reporter in HepG2, LO2, A549, and HBE cells were established. After treated with heat shock and a group of silver nanoparticles (AgNPs) with different primary particle sizes, the cell viability, oxidative damage, and luciferase activity were determined. The time-dependent Ag + ions release from AgNPs in cell medium was also evaluated. Our results showed that heat shock produced a strong time-dependent induction of relative luciferase activity in the four luciferase reporter cells. Surprisingly, at 4 h of recovery, the relative luciferase activity was > 98 × the control level in HepG2-luciferase cells. Exposure to different sizes of AgNPs resulted in activation of the HSPA1A promoter in a dose-dependent manner, even at low cytotoxic or non-cytotoxic doses. The smaller (5 nm) AgNPs were more potent in luciferase induction than the larger (50 and 75 nm) AgNPs. These results were generally in accordance with the oxidative damage indicated by malondialdehyde concentration, reactive oxygen species induction and glutathione depletion, and Ag + ions release in cell medium. Compared with the other three luciferase reporter cells, the luciferase signal in HepG2-luciferase cells is obviously more sensitive and stable. We conclude that the luciferase reporter cells, especially the HepG2-luciferase cells, could provide a valuable tool for rapid screening of the oxidative damage induced by AgNPs. - Highlights: • We established the stable HSPA1A promoter-driven luciferase reporter cells. • Silver nanoparticles induced dose-dependent increases in luciferase activity. • HSPA1A promoter activity is a sensitive and responsive indicator of oxidative stress. • HepG2-luciferase

  12. Development of HSPA1A promoter-driven luciferase reporter gene assays in human cells for assessing the oxidative damage induced by silver nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Xin, Lili, E-mail: llxin@suda.edu.cn [School of Public Health, Medical College of Soochow University, 199 Renai Road, Suzhou 215123, Jiangsu (China); Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, School of Public Health, Soochow University, Suzhou 215123 (China); Wang, Jianshu [Suzhou Center for Disease Prevention and Control, 72 Sanxiang Road, Suzhou, Jiangsu (China); Zhang, Leshuai W. [School of Radiation Medicine and Protection & School for Radiological and Interdisciplinary Sciences (RAD-X), Soochow University, 215123 (China); Che, Bizhong; Dong, Guangzhu [School of Public Health, Medical College of Soochow University, 199 Renai Road, Suzhou 215123, Jiangsu (China); Fan, Guoqiang; Cheng, Kaiming [Suzhou Industrial Park Centers for Disease Control and Prevention, 58 Suqian Road, Suzhou, Jiangsu (China)

    2016-08-01

    The exponential increase in the total number of engineered nanoparticles in consumer products requires novel tools for rapid and cost-effective toxicology screening. In order to assess the oxidative damage induced by nanoparticles, toxicity test systems based on a human HSPA1A promoter-driven luciferase reporter in HepG2, LO2, A549, and HBE cells were established. After treated with heat shock and a group of silver nanoparticles (AgNPs) with different primary particle sizes, the cell viability, oxidative damage, and luciferase activity were determined. The time-dependent Ag{sup +} ions release from AgNPs in cell medium was also evaluated. Our results showed that heat shock produced a strong time-dependent induction of relative luciferase activity in the four luciferase reporter cells. Surprisingly, at 4 h of recovery, the relative luciferase activity was > 98 × the control level in HepG2-luciferase cells. Exposure to different sizes of AgNPs resulted in activation of the HSPA1A promoter in a dose-dependent manner, even at low cytotoxic or non-cytotoxic doses. The smaller (5 nm) AgNPs were more potent in luciferase induction than the larger (50 and 75 nm) AgNPs. These results were generally in accordance with the oxidative damage indicated by malondialdehyde concentration, reactive oxygen species induction and glutathione depletion, and Ag{sup +} ions release in cell medium. Compared with the other three luciferase reporter cells, the luciferase signal in HepG2-luciferase cells is obviously more sensitive and stable. We conclude that the luciferase reporter cells, especially the HepG2-luciferase cells, could provide a valuable tool for rapid screening of the oxidative damage induced by AgNPs. - Highlights: • We established the stable HSPA1A promoter-driven luciferase reporter cells. • Silver nanoparticles induced dose-dependent increases in luciferase activity. • HSPA1A promoter activity is a sensitive and responsive indicator of oxidative stress. • HepG2

  13. Aerosol delivery of Akt controls protein translation in the lungs of dual luciferase reporter mice.

    Science.gov (United States)

    Tehrani, A M; Hwang, S-K; Kim, T-H; Cho, C-S; Hua, J; Nah, W-S; Kwon, J-T; Kim, J-S; Chang, S-H; Yu, K-N; Park, S-J; Bhandari, D R; Lee, K-H; An, G-H; Beck, G R; Cho, M-H

    2007-03-01

    Lung cancer has emerged as a leading cause of cancer death in the world; however, most of the current conventional therapies are not sufficiently effective in altering the progression of disease. Therefore, development of novel treatment approaches is needed. Although several genes and methods have been used for cancer gene therapy, a number of problems such as specificity, efficacy and toxicity reduce their application. This has led to re-emergence of aerosol gene delivery as a noninvasive method for lung cancer treatment. In this study, nano-sized glucosylated polyethyleneimine (GPEI) was used as a gene delivery carrier to investigate the effects of Akt wild type (WT) and kinase deficient (KD) on Akt-related signaling pathways and protein translation in the lungs of CMV- LucR-cMyc-IRES-LucF dual reporter mice. These mice are a powerful tool for the discrimination between cap-dependent/-independent protein translation. Aerosols containing self-assembled nano-sized GPEI/Akt WT or GPEI/Akt KD were delivered into the lungs of reporter mice through nose-only-inhalation-chamber with the aid of nebulizer. Aerosol delivery of Akt WT caused the increase of protein expression levels of Akt-related signals, whereas aerosol delivery of Akt KD did not. Furthermore, dual luciferase activity assay showed that aerosol delivery of Akt WT enhanced cap-dependent protein translation, whereas a reduction in cap-dependent protein translation by Akt KD was observed. Our results clearly showed that targeting Akt may be a good strategy for prevention as well as treatment of lung cancer. These studies suggest that our aerosol delivery is compatible for in vivo gene delivery which could be used as a noninvasive gene therapy in the future.

  14. Uptake kinetics and biodistribution of C-14-D-luciferin-a radiolabeled substrate for the firefly luciferase catalyzed bioluminescence reaction : impact on bioluminescence based reporter gene imaging

    NARCIS (Netherlands)

    Berger, Frank; Paulmurugan, Ramasamy; Bhaumik, Srabani; Gambhir, Sanjiv Sam

    2008-01-01

    Purpose Firefly luciferase catalyzes the oxidative decarboxylation of D-luciferin to oxyluciferin in the presence of cofactors, producing bioluminescence. This reaction is used in optical bioluminescence-based molecular imaging approaches to detect the expression of the firefly luciferase reporter

  15. Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression.

    Science.gov (United States)

    Puckette, Michael; Burrage, Thomas; Neilan, John G; Rasmussen, Max

    2017-06-12

    The Gaussia princeps luciferase is used as a stand-alone reporter of transgene expression for in vitro and in vivo expression systems due to the rapid and easy monitoring of luciferase activity. We sought to simultaneously quantitate production of other recombinant proteins by transcriptionally linking the Gaussia princeps luciferase gene to other genes of interest through the foot-and-mouth disease virus 2A translational interrupter sequence. We produced six plasmids, each encoding a single open reading frame, with the foot-and-mouth disease virus 2A sequence placed either N-terminal or C-terminal to the Gaussia princeps luciferase gene. Two plasmids included novel Gaussia princeps luciferase variants with the position 1 methionine deleted. Placing a foot-and-mouth disease virus 2A translational interrupter sequence on either the N- or C-terminus of the Gaussia princeps luciferase gene did not prevent the secretion or luminescence of resulting chimeric luciferase proteins. We also measured the ability of another polycistronic plasmid vector with a 2A-luciferase sequence placed downstream of the foot-and-mouth disease virus P1 and 3C protease genes to produce of foot-and-mouth disease virus-like particles and luciferase activity from transfected cells. Incorporation of the 2A-luciferase sequence into a transgene encoding foot-and-mouth disease virus structural proteins retained luciferase activity and the ability to form virus-like particles. We demonstrated a mechanism for the near real-time, sequential, non-destructive quantitative monitoring of transcriptionally-linked recombinant proteins and a valuable method for monitoring transgene expression in recombinant vaccine constructs.

  16. Buffalo Harbor Study. Preliminary Feasibility Report. Volume I. Main Report.

    Science.gov (United States)

    1983-04-01

    to usually narrow strips of riparian vegetation, which is composed of various trees and shrubs of the Salix genus (willow), sumac, aspen, boxelder...reptiles were found. Species included, leopard frogs, snapping turtles, painted turtle, and garter snakes (SUNY Brockport: 1982). (7) Endangered Species...vessel traffic on the Buffalo River causes interrupted truck service. Firms have also cited snow removal as a problem. The harbor area road service

  17. Application of the dual-luciferase reporter assay to the analysis of promoter activity in Zebrafish embryos

    Directory of Open Access Journals (Sweden)

    Mulero Victoriano

    2008-10-01

    Full Text Available Abstract Background The dual-luciferase assay has been widely used in cell lines to determine rapidly but accurately the activity of a given promoter. Although this strategy has proved very useful, it does not allow the promoter and gene function to be analyzed in the context of the whole organism. Results Here, we present a rapid and sensitive assay based on the classical dual-luciferase reporter technique which can be used as a new tool to characterize the minimum promoter region of a gene as well as the in vivo response of inducible promoters to different stimuli. We illustrate the usefulness of this system for studying both constitutive (telomerase and inducible (NF-κB-dependent promoters. The flexibility of this assay is demonstrated by induction of the NF-κB-dependent promoters using simultaneous microinjection of different pathogen-associated molecular patterns as well as with the use of morpholino-gene mediated knockdown. Conclusion This assay has several advantages compared with the classical in vitro (cell lines and in vivo (transgenic mice approaches. Among others, the assay allows a rapid and quantitative measurement of the effects of particular genes or drugs in a given promoter in the context of a whole organism and it can also be used in high throughput screening experiments.

  18. Development of functional genomic tools in trematodes: RNA interference and luciferase reporter gene activity in Fasciola hepatica.

    Directory of Open Access Journals (Sweden)

    Gabriel Rinaldi

    2008-07-01

    Full Text Available The growing availability of sequence information from diverse parasites through genomic and transcriptomic projects offer new opportunities for the identification of key mediators in the parasite-host interaction. Functional genomics approaches and methods for the manipulation of genes are essential tools for deciphering the roles of genes and to identify new intervention targets in parasites. Exciting advances in functional genomics for parasitic helminths are starting to occur, with transgene expression and RNA interference (RNAi reported in several species of nematodes, but the area is still in its infancy in flatworms, with reports in just three species. While advancing in model organisms, there is a need to rapidly extend these technologies to other parasites responsible for several chronic diseases of humans and cattle. In order to extend these approaches to less well studied parasitic worms, we developed a test method for the presence of a viable RNAi pathway by silencing the exogenous reporter gene, firefly luciferase (fLUC. We established the method in the human blood fluke Schistosoma mansoni and then confirmed its utility in the liver fluke Fasciola hepatica. We transformed newly excysted juveniles of F. hepatica by electroporation with mRNA of fLUC and three hours later were able to detect luciferase enzyme activity, concentrated mainly in the digestive ceca. Subsequently, we tested the presence of an active RNAi pathway in F. hepatica by knocking down the exogenous luciferase activity by introduction into the transformed parasites of double-stranded RNA (dsRNA specific for fLUC. In addition, we tested the RNAi pathway targeting an endogenous F. hepatica gene encoding leucine aminopeptidase (FhLAP, and observed a significant reduction in specific mRNA levels. In summary, these studies demonstrated the utility of RNAi targeting reporter fLUC as a reporter gene assay to establish the presence of an intact RNAi pathway in helminth

  19. Combined image guided monitoring the pharmacokinetics of rapamycin loaded human serum albumin nanoparticles with a split luciferase reporter

    Science.gov (United States)

    Wang, Fu; Yang, Kai; Wang, Zhe; Ma, Ying; Gutkind, J. Silvio; Hida, Naoki; Niu, Gang; Tian, Jie

    2016-02-01

    Imaging guided techniques have been increasingly employed to investigate the pharmacokinetics (PK) and biodistribution of nanoparticle based drug delivery systems. In most cases, however, the PK profiles of drugs could vary significantly from those of drug delivery carriers upon administration in the blood circulation, which complicates the interpretation of image findings. Herein we applied a genetically encoded luciferase reporter in conjunction with near infrared (NIR) fluorophores to investigate the respective PK profiles of a drug and its carrier in a biodegradable drug delivery system. In this system, a prototype hydrophobic agent, rapamycin (Rapa), was encapsulated into human serum albumin (HSA) to form HSA Rapa nanoparticles, which were then labeled with Cy5 fluorophore to facilitate the fluorescence imaging of HSA carrier. Meanwhile, we employed transgenetic HN12 cells that were modified with a split luciferase reporter, whose bioluminescence function is regulated by Rapa, to reflect the PK profile of the encapsulated agent. It was interesting to discover that there existed an obvious inconsistency of PK behaviors between HSA carrier and rapamycin in vitro and in vivo through near infrared fluorescence imaging (NIFRI) and bioluminescence imaging (BLI) after treatment with Cy5 labeled HSA Rapa. Nevertheless, HSA Rapa nanoparticles manifested favorable in vivo PK and tumor suppression efficacy in a follow-up therapeutic study. The developed strategy of combining a molecular reporter and a fluorophore in this study could be extended to other drug delivery systems to provide profound insights for non-invasive real-time evaluation of PK profiles of drug-loaded nanoparticles in pre-clinical studies.Imaging guided techniques have been increasingly employed to investigate the pharmacokinetics (PK) and biodistribution of nanoparticle based drug delivery systems. In most cases, however, the PK profiles of drugs could vary significantly from those of drug delivery

  20. Ground-water status report, Pearl Harbor area, Hawaii, 1978

    Science.gov (United States)

    Soroos, Ronald L.; Ewart, Charles J.

    1979-01-01

    Increasing demand for freshwater in Hawaii has placed heavy stress on many of the State 's basal aquifer systems. The most heavily stressed of these systems is the Pearl Harbor on Oahu. The Pearl Harbor basal aquifer supplies as much as 277 million gallons per day. Since early in this century, spring discharge has been declining while pumpage has been increasing. Total ground-water discharge has remained steady despite short-term fluctuations. Some wells show general increases in chloride concentration while others remain steady. Chloride concentrations throughout the area show no apparent increase since 1970. Basal water head maps of the Pearl Harbor area clearly reflect the natural discharge points, which are the springs located along the shore near the center of Pearl Harbor. Basal-water hydrographs show a general decline of about 0.09 foot per year. This implies depletion of storage at a rate of about 25 million gallons per day. (USGS).

  1. R/L, a double reporter mouse line that expresses luciferase gene upon Cre-mediated excision, followed by inactivation of mRFP expression.

    Science.gov (United States)

    Jia, Junshuang; Lin, Xiaolin; Lin, Xia; Lin, Taoyan; Chen, Bangzhu; Hao, Weichao; Cheng, Yushuang; Liu, Yu; Dian, Meijuan; Yao, Kaitai; Xiao, Dong; Gu, Weiwang

    2016-10-01

    The Cre/loxP system has become an important tool for the conditional gene knockout and conditional gene expression in genetically engineered mice. The applications of this system depend on transgenic reporter mouse lines that provide Cre recombinase activity with a defined cell type-, tissue-, or developmental stage-specificity. To develop a sensitive assay for monitoring Cre-mediated DNA excisions in mice, we generated Cre-mediated excision reporter mice, designated R/L mice (R/L: mRFP(monomeric red fluorescent protein)/luciferase), express mRFP throughout embryonic development and adult stages, while Cre-mediated excision deletes a loxP-flanked mRFP reporter gene and STOP sequence, thereby activating the expression of the second reporter gene luciferase, as assayed by in vivo and ex vivo bioluminescence imaging. After germ line deletion of the floxed mRFP and STOP sequence in R/L mice by EIIa-Cre mice, the resulting luciferase transgenic mice in which the loxP-mRFP-STOP-loxP cassette is excised from all cells express luciferase in all tissues and organs examined. The expression of luciferase transgene was activated in liver of RL/Alb-Cre double transgenic mice and in brain of RL/Nestin-Cre double transgenic mice when R/L reporter mice were mated with Alb-Cre mice and Nestin-Cre mice, respectively. Our findings reveal that the double reporter R/L mouse line is able to indicate the occurrence of Cre-mediated excision from early embryonic to adult lineages. Taken together, these findings demonstrate that the R/L mice serve as a sensitive reporter for Cre-mediated DNA excision both in living animals and in organs, tissues, and cells following necropsy.

  2. Knock-in Luciferase Reporter Mice for In Vivo Monitoring of CREB Activity.

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    Dmitry Akhmedov

    Full Text Available The cAMP response element binding protein (CREB is induced during fasting in the liver, where it stimulates transcription of rate-limiting gluconeogenic genes to maintain metabolic homeostasis. Adenoviral and transgenic CREB reporters have been used to monitor hepatic CREB activity non-invasively using bioluminescence reporter imaging. However, adenoviral vectors and randomly inserted transgenes have several limitations. To overcome disadvantages of the currently used strategies, we created a ROSA26 knock-in CREB reporter mouse line (ROSA26-CRE-luc. cAMP-inducing ligands stimulate the reporter in primary hepatocytes and myocytes from ROSA26-CRE-luc animals. In vivo, these animals exhibit little hepatic CREB activity in the ad libitum fed state but robust induction after fasting. Strikingly, CREB was markedly stimulated in liver, but not in skeletal muscle, after overnight voluntary wheel-running exercise, uncovering differential regulation of CREB in these tissues under catabolic states. The ROSA26-CRE-luc mouse line is a useful resource to study dynamics of CREB activity longitudinally in vivo and can be used as a source of primary cells for analysis of CREB regulatory pathways ex vivo.

  3. Rescue the failed half-ZFN by a sensitive mammalian cell-based luciferase reporter system.

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    Weifeng Zhang

    Full Text Available ZFN technology is a powerful research tool and has been used for genome editing in cells lines, animals and plants. The generation of functional ZFNs for particular targets in mammalian genome is still challenging for an average research group. The modular-assembly method is relatively fast, easy-to-practice but has a high failure rate. Some recent studies suggested that a ZFP with low binding activity might be able to form a working ZFN pair with another binding active half-ZFP. In order to unveil the potential ZFP candidates among those with low binding activities, this paper established a highly sensitive mammalian cell-based transcriptional reporter system to assess the DNA binding activities of ZFPs by inserting multiple copies of ZFN target sequence fragment (TSF of an interested gene (e. g., hPGRN or hVEGF. Our results showed that this system increased the screening sensitivity up to 50-fold and markedly amplified the differences in the binding activities between different ZFPs. We also found that the targeted chromosomal gene repair efficiency of each hPGRN or hVEGF ZFN pair was in proportion with the combination of the binding activities of the ZFL (Left zinc finger and ZFR (Right zinc finger. A hPGRN ZFR with low binding ability was able to form a biological active ZFN if combined with a hPGRN ZFL with relatively high binding ability. Lastly, site-specific genome editing by hPGRN ZFNs generated by this system was confirmed by sequencing, and the PGRN knock-out cell line showed significantly decreased cell growth compared with the control. Our system will provide a valuable tool for further optimizing the nucleases with regard to specificity and cytotoxicity.

  4. Development of a luciferase-based reporter system to monitor Bifidobacterium breve UCC2003 persistence in mice

    Directory of Open Access Journals (Sweden)

    Hill Colin

    2008-09-01

    Full Text Available Abstract Background Probiotics such as bifidobacteria have been shown to maintain a healthy intestinal microbial balance and help protect against infections. However, despite these benefits, bifidobacteria still remain poorly understood at the biochemical, physiological and especially the genetic level. Herein we describe, for the first time, the development of a non-invasive luciferase-based reporter system for real-time tracking of Bifidobacterium species in vivo. Results The reporter vector pLuxMC1 is based on the recently described theta-type plasmid pBC1 from B. catenatulatum 1 and the luxABCDE operon from pPL2lux 2. Derivatives of pLuxMC1, harbouring a bifidobacterial promoter (pLuxMC2 as well as a synthetically derived promoter (pLuxMC3 3 placed upstream of luxABCDE, were constructed and found to stably replicate in B. breve UCC2003. The subsequent analysis of these strains allowed us to assess the functionality of pLuxMC1 both in vitro and in vivo. Conclusion Our results demonstrate the potential of pLuxMC1 as a real-time, non-invasive reporter system for Bifidobacterium. It has also allowed us, for the first time, to track the colonisation potential and persistence of this probiotic species in real time. An interesting and significant outcome of the study is the identification of the caecum as a niche environment for B. breve UCC2003 within the mouse gastrointestinal tract (GI tract.

  5. Computational analysis and functional expression of ancestral copepod luciferase.

    Science.gov (United States)

    Takenaka, Yasuhiro; Noda-Ogura, Akiko; Imanishi, Tadashi; Yamaguchi, Atsushi; Gojobori, Takashi; Shigeri, Yasushi

    2013-10-10

    We recently reported the cDNA sequences of 11 copepod luciferases from the superfamily Augaptiloidea in the order Calanoida. They were classified into two groups, Metridinidae and Heterorhabdidae/Lucicutiidae families, by phylogenetic analyses. To elucidate the evolutionary processes, we have now further isolated 12 copepod luciferases from Augaptiloidea species (Metridia asymmetrica, Metridia curticauda, Pleuromamma scutullata, Pleuromamma xiphias, Lucicutia ovaliformis and Heterorhabdus tanneri). Codon-based synonymous/nonsynonymous tests of positive selection for 25 identified copepod luciferases suggested that positive Darwinian selection operated in the evolution of Heterorhabdidae luciferases, whereas two types of Metridinidae luciferases had diversified via neutral mechanism. By in silico analysis of the decoded amino acid sequences of 25 copepod luciferases, we inferred two protein sequences as ancestral copepod luciferases. They were expressed in HEK293 cells where they exhibited notable luciferase activity both in intracellular lysates and cultured media, indicating that the luciferase activity was established before evolutionary diversification of these copepod species. © 2013.

  6. Aerosol-delivered programmed cell death 4 enhanced apoptosis, controlled cell cycle and suppressed AP-1 activity in the lungs of AP-1 luciferase reporter mice.

    Science.gov (United States)

    Hwang, S-K; Jin, H; Kwon, J T; Chang, S-H; Kim, T H; Cho, C-S; Lee, K H; Young, M R; Colburn, N H; Beck, G R; Yang, H-S; Cho, M-H

    2007-09-01

    The long-term survival of lung cancer patients treated with conventional therapies remains poor and therefore the need for novel approaches remains high. This has led to the re-emergence of aerosol delivery as a therapeutic intervention. In this study, glucosylated polyethylenimine (GPEI) was used as carrier to investigate programmed cell death 4 (PDCD4) and PDCD4 mutant (D418A), an eIF4A-binding mutant, on PDCD4-related signaling and activator protein-1 (AP-1) activity in the lungs of AP-1 luciferase reporter mice. After confirming the efficiency of GPEI as a carrier in lungs, the effects of aerosol-delivered PDCD4 were investigated in AP-1 luciferase reporter mice. Aerosol delivery of GPEI/PDCD4 through a nose-only inhalation facilitated the apoptosis of lungs whereas aerosol PDCD4 mutant did not. Also, such aerosol delivery regulated proteins relevant to cell-cycle control and suppressed AP-1 activity. Results obtained by western blot analysis, immunohistochemistry, luciferase assay and deoxynucleotidyl-transferase-mediated nick end labeling study suggest that combined actions such as facilitating apoptosis, controlling cell cycle and suppression of AP-1 activity by PDCD4 may provide useful tool for designing lung tumor prevention and treatment by which PDCD4 functions as a transformation suppressor in the future.

  7. Third-generation Ah receptor-responsive luciferase reporter plasmids: amplification of dioxin-responsive elements dramatically increases CALUX bioassay sensitivity and responsiveness.

    Science.gov (United States)

    He, Guochun; Tsutsumi, Tomoaki; Zhao, Bin; Baston, David S; Zhao, Jing; Heath-Pagliuso, Sharon; Denison, Michael S

    2011-10-01

    2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related dioxin-like chemicals are widespread and persistent environmental contaminants that produce diverse toxic and biological effects through their ability to bind to and activate the Ah receptor (AhR) and AhR-dependent gene expression. The chemically activated luciferase expression (CALUX) system is an AhR-responsive recombinant luciferase reporter gene-based cell bioassay that has been used in combination with chemical extraction and cleanup methods for the relatively rapid and inexpensive detection and relative quantitation of dioxin and dioxin-like chemicals in a wide variety of sample matrices. Although the CALUX bioassay has been validated and used extensively for screening purposes, it has some limitations when screening samples with very low levels of dioxin-like chemicals or when there is only a small amount of sample matrix for analysis. Here, we describe the development of third-generation (G3) CALUX plasmids with increased numbers of dioxin-responsive elements, and stable transfection of these new plasmids into mouse hepatoma (Hepa1c1c7) cells has produced novel amplified G3 CALUX cell bioassays that respond to TCDD with a dramatically increased magnitude of luciferase induction and significantly lower minimal detection limit than existing CALUX-type cell lines. The new G3 CALUX cell lines provide a highly responsive and sensitive bioassay system for the detection and relative quantitation of very low levels of dioxin-like chemicals in sample extracts.

  8. In Vivo Determination of Vitamin D Function Using Transgenic Mice Carrying a Human Osteocalcin Luciferase Reporter Gene

    Directory of Open Access Journals (Sweden)

    Tomoko Nakanishi

    2013-01-01

    Full Text Available Vitamin D is an essential factor for ossification, and its deficiency causes rickets. Osteocalcin, which is a noncollagenous protein found in bone matrix and involved in mineralization and calcium ion homeostasis, is one of the major bone morphogenetic markers and is used in the evaluation of osteoblast maturation and osteogenic activation. We established transgenic mouse line expressing luciferase under the control of a 10-kb osteocalcin enhancer/promoter sequence. Using these transgenic mice, we evaluated the active forms of vitamins D2 and D3 for their bone morphogenetic function by in vivo bioluminescence. As the result, strong activity for ossification was observed with 1α,25-hydroxyvitamin D3. Our mouse system can offer a feasible detection method for assessment of osteogenic activity in the development of functional foods and medicines by noninvasive screening.

  9. Third-Generation Ah Receptor–Responsive Luciferase Reporter Plasmids: Amplification of Dioxin-Responsive Elements Dramatically Increases CALUX Bioassay Sensitivity and Responsiveness

    Science.gov (United States)

    He, Guochun; Tsutsumi, Tomoaki; Zhao, Bin; Baston, David S.; Zhao, Jing; Heath-Pagliuso, Sharon; Denison, Michael S.

    2011-01-01

    2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related dioxin-like chemicals are widespread and persistent environmental contaminants that produce diverse toxic and biological effects through their ability to bind to and activate the Ah receptor (AhR) and AhR-dependent gene expression. The chemically activated luciferase expression (CALUX) system is an AhR-responsive recombinant luciferase reporter gene–based cell bioassay that has been used in combination with chemical extraction and cleanup methods for the relatively rapid and inexpensive detection and relative quantitation of dioxin and dioxin-like chemicals in a wide variety of sample matrices. Although the CALUX bioassay has been validated and used extensively for screening purposes, it has some limitations when screening samples with very low levels of dioxin-like chemicals or when there is only a small amount of sample matrix for analysis. Here, we describe the development of third-generation (G3) CALUX plasmids with increased numbers of dioxin-responsive elements, and stable transfection of these new plasmids into mouse hepatoma (Hepa1c1c7) cells has produced novel amplified G3 CALUX cell bioassays that respond to TCDD with a dramatically increased magnitude of luciferase induction and significantly lower minimal detection limit than existing CALUX-type cell lines. The new G3 CALUX cell lines provide a highly responsive and sensitive bioassay system for the detection and relative quantitation of very low levels of dioxin-like chemicals in sample extracts. PMID:21775728

  10. Radiological survey of the Pearl Harbor Naval Shipyard and Environs, Honolulu, Hawaii. Final report

    International Nuclear Information System (INIS)

    Callis, R.S.

    1987-06-01

    This report presents results of the survey conducted by the Eastern Environmental Radiation Facility personnel to assess levels of environmental radioactivity resulting from maintenance and operation of nuclear-powered warships at the Pearl Harbor Naval Shipyard and Environs on the Hawaiian Island of Oahu

  11. Development and Validation of a Novel Dual Luciferase Reporter Gene Assay to Quantify Ebola Virus VP24 Inhibition of IFN Signaling

    Directory of Open Access Journals (Sweden)

    Elisa Fanunza

    2018-02-01

    Full Text Available The interferon (IFN system is the first line of defense against viral infections. Evasion of IFN signaling by Ebola viral protein 24 (VP24 is a critical event in the pathogenesis of the infection and, hence, VP24 is a potential target for drug development. Since no drugs target VP24, the identification of molecules able to inhibit VP24, restoring and possibly enhancing the IFN response, is a goal of concern. Accordingly, we developed a dual signal firefly and Renilla luciferase cell-based drug screening assay able to quantify IFN-mediated induction of Interferon Stimulated Genes (ISGs and its inhibition by VP24. Human Embryonic Kidney 293T (HEK293T cells were transiently transfected with a luciferase reporter gene construct driven by the promoter of ISGs, Interferon-Stimulated Response Element (ISRE. Stimulation of cells with IFN-α activated the IFN cascade leading to the expression of ISRE. Cotransfection of cells with a plasmid expressing VP24 cloned from a virus isolated during the last 2014 outbreak led to the inhibition of ISRE transcription, quantified by a luminescent signal. To adapt this system to test a large number of compounds, we performed it in 96-well plates; optimized the assay analyzing different parameters; and validated the system by calculating the Z′- and Z-factor, which showed values of 0.62 and 0.53 for IFN-α stimulation assay and VP24 inhibition assay, respectively, indicative of robust assay performance.

  12. Oestrogenic activity of a textile industrial wastewater treatment plant effluent evaluated by the E-screen test and MELN gene-reporter luciferase assay

    Energy Technology Data Exchange (ETDEWEB)

    Schiliro, Tiziana, E-mail: tiziana.schiliro@unito.it [Department of Public Health and Microbiology, University of Torino, Via Santena 5bis, 10126 Torino (Italy); Porfido, Arianna [Department of Public Health and Microbiology, University of Torino, Via Santena 5bis, 10126 Torino (Italy); Spina, Federica; Varese, Giovanna Cristina [Department of Life Sciences and Systems Biology, University of Torino, Viale Mattioli 25, 10125 Torino (Italy); Gilli, Giorgio [Department of Public Health and Microbiology, University of Torino, Via Santena 5bis, 10126 Torino (Italy)

    2012-08-15

    This study quantified the biological oestrogenic activity in the effluent of a textile industrial wastewater treatment plant (IWWTP) in northwestern Italy. Samples of the IWWTP effluent were collected monthly, both before and after tertiary treatment (ozonation). After solid phase extraction, all samples were subjected to two in vitro tests of total estrogenic activity, the human breast cancer cell line (MCF-7 BUS) proliferation assay, or E-screen test, and the luciferase-transfected human breast cancer cell line (MELN) gene-reporter assay, to measure the 17{beta}-oestradiol equivalent quantity (EEQ). In the E-screen test, the mean EEQ values were 2.35 {+-} 1.68 ng/L pre-ozonation and 0.72 {+-} 0.58 ng/L post-ozonation; in the MELN gene-reporter luciferase assay, the mean EEQ values were 4.18 {+-} 3.54 ng/L pre-ozonation and 2.53 {+-} 2.48 ng/L post-ozonation. These results suggest that the post-ozonation IWWTP effluent had a lower oestrogenic activity (simple paired t-tests, p < 0.05). The average reduction of estrogenic activity of IWWTP effluent after ozonation was 67 {+-} 26% and 52 {+-} 27% as measured by E-screen test and MELN gene-reporter luciferase assay, respectively. There was a positive and significant correlation between the two tests (Rho S = 0.650, p = 0.022). This study indicates that the environmental risk is low because oestrogenic substances are deposited into the river via IWWTP at concentrations lower than those at which chronic exposure has been reported to affect the endocrine system of living organisms. -- Highlights: Black-Right-Pointing-Pointer The two in vitro tests are suited for oestrogenic activity assessment in textile WWTP. Black-Right-Pointing-Pointer There is a significant correlation between the results of the two in vitro tests. Black-Right-Pointing-Pointer The oestrogenic activity of the effluent is reduced by ozonation. Black-Right-Pointing-Pointer The input of estrogenic substances into the river via textile WWTP is low.

  13. A luciferase reporter gene assay and aryl hydrocarbon receptor 1 genotype predict the LD50 of polychlorinated biphenyls in avian species

    International Nuclear Information System (INIS)

    Manning, Gillian E.; Farmahin, Reza; Crump, Doug; Jones, Stephanie P.; Klein, Jeff; Konstantinov, Alex; Potter, Dave; Kennedy, Sean W.

    2012-01-01

    Birds differ in sensitivity to the embryotoxic effects of polychlorinated biphenyls (PCBs), which complicates environmental risk assessments for these chemicals. Recent research has shown that the identities of amino acid residues 324 and 380 in the avian aryl hydrocarbon receptor 1 (AHR1) ligand binding domain (LBD) are primarily responsible for differences in avian species sensitivity to selected dibenzo-p-dioxins and furans. A luciferase reporter gene (LRG) assay was developed in our laboratory to measure AHR1-mediated induction of a cytochrome P450 1A5 reporter gene in COS-7 cells transfected with different avian AHR1 constructs. In the present study, the LRG assay was used to measure the concentration-dependent effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and PCBs 126, 77, 105 and 118 on luciferase activity in COS-7 cells transfected with AHR1 constructs representative of 86 avian species in order to predict their sensitivity to PCB-induced embryolethality and the relative potency of PCBs in these species. The results of the LRG assay indicate that the identity of amino acid residues 324 and 380 in the AHR1 LBD are the major determinants of avian species sensitivity to PCBs. The relative potency of PCBs did not differ greatly among AHR1 constructs. Luciferase activity was significantly correlated with embryolethality data obtained from the literature (R 2 ≥ 0.87, p < 0.0001). Thus, the LRG assay in combination with the knowledge of a species' AHR1 LBD sequence can be used to predict PCB-induced embryolethality in potentially any avian species of interest without the use of lethal methods on a large number of individuals. -- Highlights: ► PCB embryolethality in birds can be predicted from a species' AHR1 genotype. ► The reporter gene assay is useful for predicting species sensitivity to PCBs. ► The relative potency of PCBs does not appear to differ between AHR1 genotypes. ► Contamination of PCB 105 and PCB 118 did not affect their relative

  14. A luciferase reporter gene assay and aryl hydrocarbon receptor 1 genotype predict the LD{sub 50} of polychlorinated biphenyls in avian species

    Energy Technology Data Exchange (ETDEWEB)

    Manning, Gillian E., E-mail: gmann017@uottawa.ca [Centre for Advanced Research in Environmental Genomics, Department of Biology, University of Ottawa, Ottawa, ON, Canada K1N 6N5 (Canada); Environment Canada, National Wildlife Research Centre, Ottawa, ON, Canada K1A 0H3 (Canada); Farmahin, Reza, E-mail: mfarm070@uottawa.ca [Centre for Advanced Research in Environmental Genomics, Department of Biology, University of Ottawa, Ottawa, ON, Canada K1N 6N5 (Canada); Environment Canada, National Wildlife Research Centre, Ottawa, ON, Canada K1A 0H3 (Canada); Crump, Doug, E-mail: doug.crump@ec.gc.ca [Environment Canada, National Wildlife Research Centre, Ottawa, ON, Canada K1A 0H3 (Canada); Jones, Stephanie P., E-mail: stephanie.jones@ec.gc.ca [Environment Canada, National Wildlife Research Centre, Ottawa, ON, Canada K1A 0H3 (Canada); Klein, Jeff, E-mail: jeffery@well-labs.com [Wellington Laboratories Inc., Research Division, Guelph, ON, Canada N1G 3M5 (Canada); Konstantinov, Alex, E-mail: alex@well-labs.com [Wellington Laboratories Inc., Research Division, Guelph, ON, Canada N1G 3M5 (Canada); Potter, Dave, E-mail: dpotter@well-labs.com [Wellington Laboratories Inc., Research Division, Guelph, ON, Canada N1G 3M5 (Canada); Kennedy, Sean W., E-mail: sean.kennedy@ec.gc.ca [Centre for Advanced Research in Environmental Genomics, Department of Biology, University of Ottawa, Ottawa, ON, Canada K1N 6N5 (Canada); Environment Canada, National Wildlife Research Centre, Ottawa, ON, Canada K1A 0H3 (Canada)

    2012-09-15

    Birds differ in sensitivity to the embryotoxic effects of polychlorinated biphenyls (PCBs), which complicates environmental risk assessments for these chemicals. Recent research has shown that the identities of amino acid residues 324 and 380 in the avian aryl hydrocarbon receptor 1 (AHR1) ligand binding domain (LBD) are primarily responsible for differences in avian species sensitivity to selected dibenzo-p-dioxins and furans. A luciferase reporter gene (LRG) assay was developed in our laboratory to measure AHR1-mediated induction of a cytochrome P450 1A5 reporter gene in COS-7 cells transfected with different avian AHR1 constructs. In the present study, the LRG assay was used to measure the concentration-dependent effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and PCBs 126, 77, 105 and 118 on luciferase activity in COS-7 cells transfected with AHR1 constructs representative of 86 avian species in order to predict their sensitivity to PCB-induced embryolethality and the relative potency of PCBs in these species. The results of the LRG assay indicate that the identity of amino acid residues 324 and 380 in the AHR1 LBD are the major determinants of avian species sensitivity to PCBs. The relative potency of PCBs did not differ greatly among AHR1 constructs. Luciferase activity was significantly correlated with embryolethality data obtained from the literature (R{sup 2} ≥ 0.87, p < 0.0001). Thus, the LRG assay in combination with the knowledge of a species' AHR1 LBD sequence can be used to predict PCB-induced embryolethality in potentially any avian species of interest without the use of lethal methods on a large number of individuals. -- Highlights: ► PCB embryolethality in birds can be predicted from a species' AHR1 genotype. ► The reporter gene assay is useful for predicting species sensitivity to PCBs. ► The relative potency of PCBs does not appear to differ between AHR1 genotypes. ► Contamination of PCB 105 and PCB 118 did not affect

  15. Development and validation of cell-based luciferase reporter gene assays for measuring neutralizing anti-drug antibodies against interferon beta

    DEFF Research Database (Denmark)

    Hermanrud, Christina; Ryner, Malin; Luft, Thomas

    2016-01-01

    a normal distribution for the majority of runs, allowing a parametric approach for cut-point calculation to be used, where NAb positive samples could be identified with 95% confidence. An analysis of means and variances indicated that a floating cut-point should be used for all assays. The assays......Neutralizing anti-drug antibodies (NAbs) against therapeutic interferon beta (IFNβ) in people with multiple sclerosis (MS) are measured with cell-based bioassays. The aim of this study was to redevelop and validate two luciferase reporter-gene bioassays, LUC and iLite, using a cut-point approach...... to identify NAb positive samples. Such an approach is favored by the pharmaceutical industry and governmental regulatory agencies as it has a clear statistical basis and overcomes the limitations of the current assays based on the Kawade principle. The work was conducted following the latest assay guidelines...

  16. Development and validation of cell-based luciferase reporter gene assays for measuring neutralizing anti-drug antibodies against interferon beta.

    Science.gov (United States)

    Hermanrud, Christina; Ryner, Malin; Luft, Thomas; Jensen, Poul Erik; Ingenhoven, Kathleen; Rat, Dorothea; Deisenhammer, Florian; Sørensen, Per Soelberg; Pallardy, Marc; Sikkema, Dan; Bertotti, Elisa; Kramer, Daniel; Creeke, Paul; Fogdell-Hahn, Anna

    2016-03-01

    Neutralizing anti-drug antibodies (NAbs) against therapeutic interferon beta (IFNβ) in people with multiple sclerosis (MS) are measured with cell-based bioassays. The aim of this study was to redevelop and validate two luciferase reporter-gene bioassays, LUC and iLite, using a cut-point approach to identify NAb positive samples. Such an approach is favored by the pharmaceutical industry and governmental regulatory agencies as it has a clear statistical basis and overcomes the limitations of the current assays based on the Kawade principle. The work was conducted following the latest assay guidelines. The assays were re-developed and validated as part of the "Anti-Biopharmaceutical Immunization: Prediction and analysis of clinical relevance to minimize the risk" (ABIRISK) consortium and involved a joint collaboration between four academic laboratories and two pharmaceutical companies. The LUC assay was validated at Innsbruck Medical University (LUCIMU) and at Rigshospitalet (LUCRH) Copenhagen, and the iLite assay at Karolinska Institutet, Stockholm. For both assays, the optimal serum sample concentration in relation to sensitivity and recovery was 2.5% (v/v) in assay media. A Shapiro-Wilk test indicated a normal distribution for the majority of runs, allowing a parametric approach for cut-point calculation to be used, where NAb positive samples could be identified with 95% confidence. An analysis of means and variances indicated that a floating cut-point should be used for all assays. The assays demonstrated acceptable sensitivity for being cell-based assays, with a confirmed limit of detection in neat serum of 1519 ng/mL for LUCIMU, 814 ng/mL for LUCRH, and 320 ng/mL for iLite. Use of the validated cut-point assay, in comparison with the previously used Kawade method, identified 14% more NAb positive samples. In conclusion, implementation of the cut-point design resulted in increased sensitivity to detect NAbs. However, the clinical significance of these low

  17. Cellular Immune Response Against Firefly Luciferase After Sleeping Beauty–Mediated Gene Transfer In Vivo

    Science.gov (United States)

    Podetz-Pedersen, Kelly M.; Vezys, Vaiva; Somia, Nikunj V.; Russell, Stephen J.

    2014-01-01

    Abstract The Sleeping Beauty (SB) transposon system has been shown to mediate new gene sequence integration resulting in long-term expression. Here the effectiveness of hyperactive SB100X transposase was tested, and we found that hydrodynamic co-delivery of a firefly luciferase transposon (pT2/CaL) along with SB100X transposase (pCMV-SB100X) resulted in remarkably sustained, high levels of luciferase expression. However, after 4 weeks there was a rapid, animal-by-animal loss of luciferase expression that was not observed in immunodeficient mice. We hypothesized that this sustained, high-level luciferase expression achieved using the SB100X transposase elicits an immune response in pT2/CaL co-administered mice, which was supported by the rapid loss of luciferase expression upon challenge of previously treated animals and in naive animals adoptively transferred with splenocytes from previously treated animals. Specificity of the immune response to luciferase was demonstrated by increased cytokine expression in splenocytes after exposure to luciferase peptide in parallel with MHC I–luciferase peptide tetramer binding. This anti-luciferase immune response observed following continuous, high-level luciferase expression in vivo clearly impacts its use as an in vivo reporter. As both an immunogen and an extremely sensitive reporter, luciferase is also a useful model system for the study of immune responses following in vivo gene transfer and expression. PMID:25093708

  18. Tibetan Firefly Luciferase with Low Temperature Adaptation.

    Science.gov (United States)

    Mitani, Yasuo; Futahashi, Ryo; Liu, Zichao; Liang, Xingcai; Ohmiya, Yoshihiro

    2017-03-01

    Fireflies are widespread all over the world and a numerous numbers of luciferases have been isolated and characterized. In this study, we identified and characterized the luciferase and luciferase-like genes from a Tibetan firefly collected in Shangri-La, China. The altitude of this area is more than 3300 m. We saw this Tibetan firefly flying with strong luminescence after sunset at ~10°C. We analyzed the transcriptome of Tibetan firefly using head, thorax, abdomen (without light organ), and light organ tissue by RNA sequencing. We identified one luciferase gene, which was almost identical to luciferase from fireflies Pyrocoelia species, and expressed specifically in the light organ. Interestingly, the optimal temperature of the Tibetan firefly recombinant luciferase was 10°C. The K m for D-luciferin and ATP of the recombinant luciferase was 23 and 154 μm, respectively. The optimal pH was around 7.0-7.5. The emission peak was 556 nm at pH 8.0, while it shifted to 606 nm at pH 6.0. We also found a luciferase-like gene with 43% identical amino acids to the Tibetan firefly luciferase, which was scarcely expressed in any portion of the adult body. No luciferase activity was detected for this luciferase-like protein. © 2016 The American Society of Photobiology.

  19. Space-time analysis of gravitropism in etiolated Arabidopsis hypocotyls using bioluminescence imaging of the IAA19 promoter fusion with a destabilized luciferase reporter.

    Science.gov (United States)

    Yamamoto, Kotaro T; Watahiki, Masaaki K; Matsuzaki, Jun; Satoh, Soichirou; Shimizu, Hisayo

    2017-07-01

    Imaging analysis was carried out during the gravitropic response of etiolated Arabidopsis hypocotyls, using an IAA19 promoter fusion of destabilized luciferase as a probe. From the bright-field images we obtained the local deflection angle to the vertical, A, local curvature, C, and the partial derivative of C with respect to time, [Formula: see text]. These were determined every 19.9 µm along the curvilinear length of the hypocotyl, at ~10 min intervals over a period of ~6 h after turning hypocotyls through 90° to the horizontal. Similarly from the luminescence images we measured the luminescence intensity of the convex and concave flanks of the hypocotyl as well as along the median of the hypocotyl, to determine differential expression of auxin-inducible IAA19. Comparison of these parameters as a function of time and curvilinear length shows that the gravitropic response is composed of three successive elements: the first and second curving responses and a decurving response (autostraightening). The maximum of the first curving response occurs when A is 76° along the entire length of the hypocotyl, suggesting that A is the sole determinant in this response; in contrast, the decurving response is a function of both A and C, as predicted by the newly-proposed graviproprioception model (Bastien et al., Proc Natl Acad Sci USA 110:755-760, 2013). Further, differential expression of IAA19, with higher expression in the convex flank, is observed at A = 44°, and follows the Sachs' sine law. This also suggests that IAA19 is not involved in the first curving response. In summary, the gravitropic response of Arabidopsis hypocotyls consists of multiple elements that are each determined by separate principles.

  20. Luciferase assay to study the activity of a cloned promoter DNA fragment.

    Science.gov (United States)

    Solberg, Nina; Krauss, Stefan

    2013-01-01

    Luciferase based assays have become an invaluable tool for the analysis of cloned promoter DNA fragments, both for verifying the ability of a potential promoter fragment to drive the expression of a luciferase reporter gene in various cellular contexts, and for dissecting binding elements in the promoter. Here, we describe the use of the Dual-Luciferase(®) Reporter Assay System created by Promega (Promega Corporation, Wisconsin, USA) to study the cloned 6.7 kilobases (kb) mouse (m) Tcf3 promoter DNA fragment in mouse embryonic derived neural stem cells (NSC). In this system, the expression of the firefly luciferase driven by the cloned mTcf3 promoter DNA fragment (including transcription initiation sites) is correlated with a co-transfected control reporter expressing Renilla luciferase from the herpes simplex virus (HSV) thymidine kinase promoter. Using an internal control reporter allows to normalize the activity of the experimental reporter to the internal control, which minimizes experimental variability.

  1. [Pearl Harbor.

    Science.gov (United States)

    Johnson, Jennifer, Ed.

    1992-01-01

    This issue of "Loblolly Magazine" was written in observance of the 50th anniversary of the U.S. entrance into World War II. The publication features interviews conducted by East Texas high school students with Clarence Otterman, one of the few survivors of the crew of the USS Arizona, which was bombed during the attack on Pearl Harbor,…

  2. A marked response to icotinib in a patient with large cell neuroendocrine carcinoma harboring an EGFR mutation: A case report.

    Science.gov (United States)

    Wang, Yuehong; Shen, Yi Hong; Ma, Shanni; Zhou, Jianying

    2015-09-01

    The present study reports the case of an 84-year-old male with primary pulmonary large cell neuroendocrine carcinoma (LCNEC) harboring an epidermal growth factor receptor (EGFR) gene mutation that exhibited a long-lasting response to the EGFR-tyrosine kinase inhibitor (EGFR-TKI) icotinib. The patient had an extensive smoking history, a poor performance status, and presented with an irregular mass in the middle lobe of the right lung on computed tomography (CT) and an enlarged left supraclavicular lymph node on physical examination. Right middle lobe bronchial brushing during fiberoptic bronchoscopy identified poorly-differentiated cancer cells. The left supraclavicular lymph node was biopsied and a diagnosis of metastatic LCNEC was determined. Furthermore, an EGFR exon 19 deletion was identified by DNA sequencing. Following diagnosis, icotinib was administered at a dose of 125 mg three times a day. Chest CT scans were performed after 1 month of treatment, which indicated that the tumor was in partial remission. This marked response to icotinib lasted for 8 months. Thus, the present case illustrates the possibility of identifying EGFR mutations in LCNEC and indicates that EGFR-tyrosine kinase inhibitors may be an alternative treatment strategy for patients with LCNEC harboring activating EGFR mutations.

  3. [Effectiveness of azacitidine in chronic myelomonocytic leukemia harboring del(20q) - a case report].

    Science.gov (United States)

    Manabe, Masahiro; Okita, Junya; Takakuwa, Teruhito; Harada, Naonori; Aoyama, Yasutaka; Kumura, Takeo; Ohta, Tadanobu; Furukawa, Yoshio; Mugitani, Atsuko

    2014-06-01

    A 7 1-year-old man was admitted to our hospital with leukocytosis and anemia. Chronic myelomonocytic leukemia (CMML)harboring del(20q)was diagnosed by peripheral blood examination and bone marrow aspiration. The patient was subsequently treated with azacitidine, which resulted in rapid disappearance of monocytosis and resolved his dependency on red cell transfusion. With regard to the chromosomal abnormality, although del(20q)is estimated to be encountered in approximately 0.7-1.0% of all CMML cases, its significance in prognosis has not been fully analyzed. Hence, more such cases need to be evaluated to elucidate the therapeutic outcome of CMML involving del(20q). In addition, the Wilms tumor-1(WT 1)level in the patient gradually decreased after the initiation of azacitidine therapy. This phenomenon of WT1 decrease synchronizing with the patient's clinical improvement might reflect therapeutic efficacy with regard to the clinical course, as had been observed in acute myeloid leukemia and myelodysplastic syndrome.

  4. Improvement of thermostability and activity of firefly luciferase through [TMG][Ac] ionic liquid mediator.

    Science.gov (United States)

    Ebrahimi, Mehdi; Hosseinkhani, Saman; Heydari, Akbar; Khavari-Nejad, Ramazan Ali; Akbari, Jafar

    2012-10-01

    Firefly luciferase catalyzes production of light from luciferin in the presence of Mg(2+)-ATP and oxygen. This enzyme has wide range of applications in biotechnology and development of biosensors. The low thermal stability of wild-type firefly luciferase is a limiting factor in most applications. Improvements in activity and stability of few enzymes in the presence of ionic liquids were shown in many reports. In this study, kinetic and thermal stability of firefly luciferase from Photinus pyralis in the presence of three tetramethylguanidine-based ionic liquids was investigated. The enzyme has shown improved activity in the presence of [1, 1, 3, 3-tetramethylguanidine][acetate], but in the presence of [TMG][trichloroacetate] and [TMG][triflouroacetate] activity, it decreased or unchanged significantly. Among these ionic liquids, only [TMG][Ac] has increased the thermal stability of luciferase. Incubation of [TMG][Ac] with firefly luciferase brought about with decrease of K(m) for ATP.

  5. Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase

    International Nuclear Information System (INIS)

    Yoshida, Wataru; Kezuka, Aki; Murakami, Yoshiyuki; Lee, Jinhee; Abe, Koichi; Motoki, Hiroaki; Matsuo, Takafumi; Shimura, Nobuaki; Noda, Mamoru; Igimi, Shizunobu; Ikebukuro, Kazunori

    2013-01-01

    Graphical abstract: -- Highlights: •Zif268 fused to luciferase was used for E. coli O157, Salmonella and coliform detection. •Artificial zinc finger protein fused to luciferase was constructed for Norovirus detection. •An analyzer that automatically detects PCR products by zinc finger protein fused to luciferase was developed. •Target pathogens were specifically detected by the automatic analyzer with zinc finger protein fused to luciferase. -- Abstract: An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268–luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF–luciferase fusion protein. By means of the automatic analyzer with ZF–luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0 × 10 to 1.0 × 10 6 copies

  6. Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase

    Energy Technology Data Exchange (ETDEWEB)

    Yoshida, Wataru; Kezuka, Aki; Murakami, Yoshiyuki; Lee, Jinhee; Abe, Koichi [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 (Japan); Motoki, Hiroaki; Matsuo, Takafumi; Shimura, Nobuaki [System Instruments Co., Ltd., 776-2 Komiya-cho, Hachioji, Tokyo 192-0031 (Japan); Noda, Mamoru; Igimi, Shizunobu [Division of Biomedical Food Research, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501 (Japan); Ikebukuro, Kazunori, E-mail: ikebu@cc.tuat.ac.jp [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 (Japan)

    2013-11-01

    Graphical abstract: -- Highlights: •Zif268 fused to luciferase was used for E. coli O157, Salmonella and coliform detection. •Artificial zinc finger protein fused to luciferase was constructed for Norovirus detection. •An analyzer that automatically detects PCR products by zinc finger protein fused to luciferase was developed. •Target pathogens were specifically detected by the automatic analyzer with zinc finger protein fused to luciferase. -- Abstract: An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268–luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF–luciferase fusion protein. By means of the automatic analyzer with ZF–luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0 × 10 to 1.0 × 10{sup 6} copies.

  7. A Luciferase Reporter Gene Assay to Measure Ebola Virus Viral Protein 35-Associated Inhibition of Double-Stranded RNA-Stimulated, Retinoic Acid-Inducible Gene 1-Mediated Induction of Interferon β.

    Science.gov (United States)

    Cannas, Valeria; Daino, Gian Luca; Corona, Angela; Esposito, Francesca; Tramontano, Enzo

    2015-10-01

    During Ebola virus (EBOV) infection, the type I interferon α/β (IFN-α/β) innate immune response is suppressed by EBOV viral protein 35 (VP35), a validated drug target. Identification of EBOV VP35 inhibitors requires a cellular system able to assess the VP35-based inhibitory functions of viral double-stranded RNA (dsRNA) IFN-β induction. We established a miniaturized luciferase gene reporter assay in A549 cells that measures IFN-β induction by viral dsRNA and is dose-dependently inhibited by VP35 expression. When compared to influenza A virus NS1 protein, EBOV VP35 showed improved inhibition of viral dsRNA-based IFN-β induction. This assay can be used to screen for EBOV VP35 inhibitors. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  8. Sensitive detection of viable Escherichia coli O157:H7 from foods using a luciferase-reporter phage phiV10lux.

    Science.gov (United States)

    Kim, Jinwoo; Kim, Minsik; Kim, Seongmi; Ryu, Sangryeol

    2017-08-02

    Escherichia coli O157:H7, a major foodborne pathogen, is a major public health concern associated with life-threatening diseases such as hemolytic uremic syndrome. To alleviate this burden, a sensitive and rapid system is required to detect this pathogen in various kinds of foods. Herein, we propose a phage-based pathogen detection method to replace laborious and time-consuming conventional methods. We engineered an E. coli O157:H7-specific phage phiV10 to rapidly and sensitively detect this notorious pathogen. The luxCDABE operon was introduced into the phiV10 genome and allowed the engineered phage phiV10lux to generate bioluminescence proportional to the number of viable E. coli O157:H7 cells without any substrate addition. The phage phiV10lux was able to detect at least 1CFU/ml of E. coli O157:H7 in a pure culture within 40min after 5h of pre-incubation. In artificially contaminated romaine lettuce, apple juice (pH3.51), and ground beef, the reporter phage could detect approximately 10CFU/cm 2 , 13CFU/ml, and 17CFU/g of E. coli O157:H7, respectively. Taken together, the constructed reporter phage phiV10lux could be applied as a powerful tool for rapid and sensitive detection of live E. coli O157:H7 in foods. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Canaveral Harbor, Florida, Integrated Section 203 Navigation Study Report and Final Environmental Assessment. Volume 1

    Science.gov (United States)

    2012-10-01

    Bielsa et al., 1983) where adult females lay demersal eggs. Spawning takes place year round in some areas (e.g., Tortugas Shelf), but peak spawning...Manatees rarely venture into deep ocean waters. However, there are reports of manatees in locations as far offshore as the Dry Tortugas Islands

  10. Interim Feasibility Report and Draft Environmental Impact Statement, Grays Harbor, Chehalis and Hoquiam Rivers, Washington, Channel Improvements for Navigation.

    Science.gov (United States)

    1982-06-01

    community depends on the continued descent of organic materials from the overlying waters for nourishment. The Dungeness crab ( Cancer magister) is an...immediately north and south of Grays Harbor. EIS-34 o Crabs. Dungeness crabs ( Cancer magister) are abundant in Grays Harbor, though most are smaller...Cooper Cultural Resources Mama Seattle District, Evaluation Corps of Engineers Endangered Species Brunner Seattle District, Evaluation Corps of Engineers

  11. Rapid assessment of repair of ultraviolet DNA damage with a modified host-cell reactivation assay using a luciferase reporter gene and correlation with polymorphisms of DNA repair genes in normal human lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Qiao Yawei; Spitz, Margaret R.; Guo Zhaozheng; Hadeyati, Mohammad; Grossman, Lawrence; Kraemer, Kenneth H.; Wei Qingyi

    2002-11-30

    As DNA repair plays an important role in genetic susceptibility to cancer, assessment of the DNA repair phenotype is critical for molecular epidemiological studies of cancer. In this report, we compared use of the luciferase (luc) reporter gene in a host-cell reactivation (HCR) (LUC) assay of repair of ultraviolet (UV) damage to DNA to use of the chloramphenicol (cat) gene-based HCR (CAT) assay we used previously for case-control studies. We performed both the assays on cryopreserved lymphocytes from 102 healthy non-Hispanic white subjects. There was a close correlation between DNA repair capacity (DRC) as measured by the LUC and CAT assays. Although these two assays had similar variation, the LUC assay was faster and more sensitive. We also analyzed the relationship between DRC and the subjects' previously determined genotypes for four polymorphisms of two nucleotide-excision repair (NER) genes (in intron 9 of xeroderma pigmentosum (XP) C and exons 6, 10 and 23 of XPD) and one polymorphism of a base-excision repair gene in exon 10 of X-ray complementing group 1 (XRCC1). The DRC was significantly lower in subjects homozygous for one or more polymorphisms of the two NER genes than in subjects with other genotypes (P=0.010). In contrast, the polymorphic XRCC1 allele had no significant effect on DRC. These results suggest that the post-UV LUC assay measures NER phenotype and that polymorphisms of XPC and XPD genes modulate DRC. For population studies of the DNA repair phenotype, many samples need to be evaluated, and so the LUC assay has several advantages over the CAT assay: the LUC assay was more sensitive, had less variation, was not radioactive, was easier to perform, and required fewer cryopreserved cells. These features make the LUC-based HCR assay suitable for molecular epidemiological studies.

  12. Fe65 does not stabilize AICD during activation of transcription in a luciferase assay

    International Nuclear Information System (INIS)

    Huysseune, Sandra; Kienlen-Campard, Pascal; Octave, Jean-Noel

    2007-01-01

    The APP intracellular domain (AICD) could be involved in signaling via interaction with the adaptor protein Fe65, and with the histone acetyl transferase Tip60. However, the real function of AICD and Fe65 in regulation of transcription remains controversial. In this study, the human APPGal4 fusion protein was expressed in CHO cells and the transcriptional activity of AICDGal4 was measured in a luciferase-based reporter assay. AICDGal4 was stabilized by expression of Fe65 and levels of AICDGal4 controlled luciferase activity. On the contrary, when human APP was expressed in CHO cells, coexpression of Fe65 increased luciferase activity without affecting the amount of AICD fragment. AICD produced from APP was protected from degradation by orthophenanthroline, but not by lactacystine, indicating that AICD is not a substrate of the chymotryptic activity of the proteasome. It is concluded that Fe65 can control luciferase activity without stabilizing the labile AICD fragment

  13. Re-engineering of Bacterial Luciferase; For New Aspects of Bioluminescence.

    Science.gov (United States)

    Kim, Da-Som; Choi, Jeong-Ran; Ko, Jeong-Ae; Kim, Kangmin

    2018-01-01

    Bacterial luminescence is the end-product of biochemical reactions catalyzed by the luciferase enzyme. Nowadays, this fascinating phenomenon has been widely used as reporter and/or sensors to detect a variety of biological and environmental processes. The enhancement or diversification of the luciferase activities will increase the versatility of bacterial luminescence. Here, to establish the strategy for luciferase engineering, we summarized the identity and relevant roles of key amino acid residues modulating luciferase in Vibrio harveyi, a model luminous bacterium. The current opinions on crystal structures and the critical amino acid residues involved in the substrate binding sites and unstructured loop have been delineated. Based on these, the potential target residues and/or parameters for enzyme engineering were also suggested in limited scale. In conclusion, even though the accurate knowledge on the bacterial luciferase is yet to be reported, the structure-guided site-directed mutagenesis approaches targeting the regulatory amino acids will provide a useful platform to re-engineer the bacterial luciferase in the future. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  14. Firefly luciferase gene contains a cryptic promoter

    Czech Academy of Sciences Publication Activity Database

    Vopálenský, V.; Mašek, T.; Horváth, Ondřej; Vicenová, B.; Mokrejš, M.; Pospíšek, M.

    2008-01-01

    Roč. 14, č. 9 (2008), s. 1720-1729 ISSN 1355-8382 Grant - others:GAČR(CZ) GA204/03/1487; GAČR(CZ) GA301/07/0607; Mšk(CZ) LC06066 Program:LC Institutional research plan: CEZ:AV0Z50520514 Keywords : luciferase * cryptic promoter * hepatitis C virus Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.018, year: 2008

  15. Pearl Harbor Biological Survey

    Science.gov (United States)

    1974-08-30

    Pearl Harbor also receives Irrigation tailgate waters from the Oahu Sug- ar Company, Industrial waste waters from the Prlmo Brewery , and heated waters...34Observations of the Cell Structure of Salt Fingers", J. Fluid Mech. 41:4, pp 707-719. ~) 3.3-79 ,’:•.-. ^ IV s’V- EFFECTS OF SHIP ACTIVITY Paul L...anticyclonic) death assemblage - in this report, an assemblage (q.v.) of remains (such as shells or bones ) from a naturally occurring association of living

  16. Coastal Harbors Modeling Facility

    Data.gov (United States)

    Federal Laboratory Consortium — The Coastal Harbors Modeling Facility is used to aid in the planning of harbor development and in the design and layout of breakwaters, absorbers, etc.. The goal is...

  17. Use of luciferase probes to measure ATP in living cells and animals.

    Science.gov (United States)

    Morciano, Giampaolo; Sarti, Alba Clara; Marchi, Saverio; Missiroli, Sonia; Falzoni, Simonetta; Raffaghello, Lizzia; Pistoia, Vito; Giorgi, Carlotta; Di Virgilio, Francesco; Pinton, Paolo

    2017-08-01

    ATP, the energy exchange factor that connects anabolism and catabolism, is required for major reactions and processes that occur in living cells, such as muscle contraction, phosphorylation and active transport. ATP is also the key molecule in extracellular purinergic signaling mechanisms, with an established crucial role in inflammation and several additional disease conditions. Here, we describe detailed protocols to measure the ATP concentration in isolated living cells and animals using luminescence techniques based on targeted luciferase probes. In the presence of magnesium, oxygen and ATP, the protein luciferase catalyzes oxidation of the substrate luciferin, which is associated with light emission. Recombinantly expressed wild-type luciferase is exclusively cytosolic; however, adding specific targeting sequences can modify its cellular localization. Using this strategy, we have constructed luciferase chimeras targeted to the mitochondrial matrix and the outer surface of the plasma membrane. Here, we describe optimized protocols for monitoring ATP concentrations in the cytosol, mitochondrial matrix and pericellular space in living cells via an overall procedure that requires an average of 3 d. In addition, we present a detailed protocol for the in vivo detection of extracellular ATP in mice using luciferase-transfected reporter cells. This latter procedure may require up to 25 d to complete.

  18. Development and application of hepatitis C reporter viruses with genotype 1 to 7 core-nonstructural protein 2 (NS2) expressing fluorescent proteins or luciferase in modified JFH1 NS5A

    DEFF Research Database (Denmark)

    Gottwein, Judith M; Jensen, Tanja B; Mathiesen, Christian K

    2011-01-01

    to 2a(J6) tagged with EGFP, DsRed-Express2, mCherry, or Renilla luciferase (RLuc), yielding peak supernatant infectivity titers of 4 to 5 log(10) focus-forming units (FFU)/ml. 2a(J6) with ¿40 or ¿25 was fully viable in Huh7.5 cells. In human liver chimeric mice, 2a(J6)-EGFP¿40 acquired various...... deletions in EGFP, while 2a(J6)¿40 did not show an impaired viability. We further developed panels of JFH1-based genotype 1 to 7 core-NS2 recombinants expressing EGFP- or RLuc-NS5A¿40 fusion proteins. In cell culture, the different EGFP recombinants showed growth characteristics comparable to those...

  19. An investigation of the feasibility of building a harbor on the West coast of South America using explosive power of nuclear weapons, a preliminary report

    Energy Technology Data Exchange (ETDEWEB)

    Zodtner, H. H.

    1971-12-31

    There is an interest in discovering the various peace time uses of nuclear explosives. One of the proposals is the building of harbors. There are several ports along the west coast of South America where lighterage is necessary. This implies a need for expanded harbor facilities. The problem is to find a good location for creating a harbor, and the feasibility of accomplishing this with the use of nuclear force. Feasibility includes blast effects, radiation hazards, the number of weapons needed, and economic considerations. Economic considerations include the cost of treating a harbor of sufficient depth and area, the building of harbor facilities, and the estimated savings and advantages of the new harbor. Several meetings were held with naval personnel of the Military Liaison group at UCRL to discuss the general problems of harbors. Thirty-three different ports were given a preliminary investigation.

  20. Three-year summary report of biological monitoring at the Southwest Ocean dredged-material disposal site and additional locations off Grays Harbor, Washington, 1990--1992

    Energy Technology Data Exchange (ETDEWEB)

    Antrim, L.D.; Shreffler, D.K.; Pearson, W.H.; Cullinan, V.I. [Battelle Marine Research Lab., Sequim, WA (United States)

    1992-12-01

    The Grays Harbor Navigation Improvement Project was initiated to improve navigation by widening and deepening the federal channel at Grays Harbor. Dredged-material disposal sites were selected after an extensive review process that included inter-agency agreements, biological surveys, other laboratory and field studies, and preparation of environmental impact statements The Southwest Site, was designated to receive materials dredged during annual maintenance dredging as well as the initial construction phase of the project. The Southwest Site was located, and the disposal operations designed, primarily to avoid impacts to Dungeness crab. The Final Environmental Impact Statement Supplement for the project incorporated a Site Monitoring Plan in which a tiered approach to disposal site monitoring was recommended. Under Tier I of the Site Monitoring Plan, Dungeness crab densities are monitored to confirm that large aggregations of newly settled Dungeness crab have not moved onto the Southwest Site. Tier 2 entails an increased sampling effort to determine whether a change in disposal operations is needed. Four epibenthic surveys using beam trawls were conducted in 1990, 1991, and 1992 at the Southwest Site and North Reference area, where high crab concentrations were found in the spring of 1985. Survey results during these three years prompted no Tier 2 activities. Epibenthic surveys were also conducted at two nearshore sites where construction of sediment berms has been proposed. This work is summarized in an appendix to this report.

  1. A transgenic Plasmodium falciparum NF54 strain that expresses GFP-luciferase throughout the parasite life cycle.

    Science.gov (United States)

    Vaughan, Ashley M; Mikolajczak, Sebastian A; Camargo, Nelly; Lakshmanan, Viswanathan; Kennedy, Mark; Lindner, Scott E; Miller, Jessica L; Hume, Jen C C; Kappe, Stefan H I

    2012-12-01

    Plasmodium falciparum is the pathogenic agent of the most lethal of human malarias. Transgenic P. falciparum parasites expressing luciferase have been created to study drug interventions of both asexual and sexual blood stages but luciferase-expressing mosquito stage and liver stage parasites have not been created which has prevented the easy quantification of mosquito stage development (e.g. for transmission blocking interventions) and liver stage development (for interventions that prevent infection). To overcome this obstacle, we have created a transgenic P. falciparum NF54 parasite that expresses a GFP-luciferase transgene throughout the life cycle. Luciferase expression is robust and measurable at all life cycle stages, including midgut oocyst, salivary gland sporozoites and liver stages, where in vivo development is easily measurable using humanized mouse infections in conjunction with an in vivo imaging system. This parasite reporter strain will accelerate testing of interventions against pre-erythrocytic life cycle stages. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. Pascagoula Harbor, Mississippi. Feasibility Report on Improvement of the Federal Deep-Draft Navigation Channel. Volume 2. Technical Appendices. Revision.

    Science.gov (United States)

    1985-03-01

    restrittd to as small an area as feasIble. Bosed on previous dredginp/disposal ;i. 1 Ions at Pascapulla lfarhr, it is ielt that in reasonable mixing...likely to adversely affect l.stcd species, the formal consu!tatlon process shall be Ini- tiated by writing to th:e Pegiona Director at _he , ddresv on th...area, plants, labor (botanist time), and maintenance, are figured on a per acre basis. Report writing , secretarial time, etc., are variable work items

  3. Luciferase inactivation in the luminous marine bacterium Vibrio harveyi.

    Science.gov (United States)

    Reeve, C A; Baldwin, T O

    1981-06-01

    Luciferase was rapidly inactivated in stationary-phase cultures of the wild type of the luminous marine bacterium Vibrio harveyi, but was stable in stationary-phase cultures of mutants of V. harveyi that are nonluminous without exogenous aldehyde, termed the aldehyde-deficient mutants. The inactivation in the wild type was halted by cell lysis and was slowed or stopped by O2 deprivation or by addition of KCN and NaF or of chloramphenicol. If KCN and NaF or chloramphenicol were added to a culture before the onset of luciferase inactivation, then luciferase inactivation did not occur. However, if these inhibitors were added after the onset of luciferase inactivation, then luciferase inactivation continued for about 2 to 3 h before the inactivation process stopped. The onset of luciferase inactivation in early stationary-phase cultures of wild-type cell coincided with a slight drop in the intracellular adenosine 5'-triphosphate (ATP) level from a relatively constant log-phase value of 20 pmol of ATP per microgram of soluble cell protein. Addition of KCN and NaF to a culture shortly after this drop in ATP caused a rapid decrease in the ATP level to about 4 pmol of ATP per microgram whereas chloramphenicol added at this same time caused a transient increase in ATP level to about 25 pmol/microgram. The aldehyde-deficient mutant (M17) showed a relatively constant log-phase ATP level identical with that of the wild-type cells, but rather than decreasing in early stationary phase, the ATP level increased to a value twice that in log-phase cells. We suggest that the inactivation of luciferase is dependent on the synthesis of some factor which is produced during stationary phase and is itself unstable, and whose synthesis is blocked by chloramphenicol or cyanide plus fluoride.

  4. Super RLuc8: A novel engineered Renilla luciferase with a red-shifted spectrum and stable light emission.

    Science.gov (United States)

    Rahnama, Somaieh; Saffar, Behnaz; Kahrani, Zahra Fanaei; Nazari, Mahboobeh; Emamzadeh, Rahman

    2017-01-01

    Renilla luciferase is a bioluminescent enzyme which is broadly used as a reporter protein in molecular biosensors. In this study, a novel luciferase with desired light emission wavelength and thermostability is reported. The results indicated that the new luciferase, namely super RLuc8, had a red-shifted spectrum and showed stable light emission. Super RLuc8 showed a 10-fold (p-value=0.0084) increase in the thermostability at 37°C after 20min incubation, in comparison to the native enzyme. The optimum temperature of the mutant increased from 30 to 37°C. Molecular dynamics simulation analysis indicated that the increased thermostability was most probably caused by a better structural compactness and more local rigidity in the regions out of the emitter site. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. The disulfide-rich Metridia luciferase refolded from E. coli inclusion bodies reveals the properties of a native folded enzyme produced in insect cells.

    Science.gov (United States)

    Markova, Svetlana V; Larionova, Marina D; Gorbunova, Darya A; Vysotski, Eugene S

    2017-10-01

    The bioluminescence of a marine copepod Metridia longa is determined by a small secreted coelenterazine-dependent luciferase that uses coelenterazine as a substrate of enzymatic reaction to generate light (λ max =480nm). To date, four different isoforms of the luciferase differing in size, sequences, and properties have been cloned by functional screening. All of them contain ten conserved Cys residues that suggests up to five SS intramolecular bonds per luciferase molecule. Whereas the use of copepod luciferases as bioluminescent reporters in biomedical research in vivo is growing from year to year, their application for in vitro assays is still limited by the difficulty in obtaining significant amounts of luciferase. The most cost-effective host for producing recombinant proteins is Escherichia coli. However, prokaryotic and eukaryotic cells maintain the reductive environment in cytoplasm that hinders the disulfide bond formation and consequently the proper folding of luciferase. Here we report the expression of the MLuc7 isoform of M. longa luciferase in E. coli cells and the efficient procedure for refolding from inclusion bodies yielding a high-active monomeric protein. Furthermore, in a set of identical experiments we demonstrate that bioluminescent and structural features of MLuc7 produced in bacterial cells are identical to those of MLuc7 isoform produced from culture medium of insect cells. Although the yield of high-purity protein is only 6mg/L, the application of E. coli cells to produce the luciferase is simpler and more cost-effective than the use of insect cells. We expect that the suggested technology of Metridia luciferase production allows obtaining of sufficient amounts of protein both for the development of novel in vitro analytical assays with the use of MLuc7 as a label and for structural studies. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Initiation of protein synthesis by a labeled derivative of the Lactobacillus casei DN-114 001 strain during transit from the stomach to the cecum in mice harboring human microbiota.

    Science.gov (United States)

    Oozeer, R; Mater, D D G; Goupil-Feuillerat, N; Corthier, G

    2004-12-01

    Although studies on the survival of bacteria in the digestive tract have been reported in the literature, little data are available on the physiological adaptation of probiotics to the digestive environment. In previous work, a transcriptional fusion system (i.e., luciferase genes under the control of a deregulated promoter) was used to demonstrate that a derivative of the Lactobacillus casei DN-114 001 strain, ingested in a fermented milk and thus exhibiting initially a very weak metabolic activity, synthesized proteins de novo after its transit in the digestive tract of mice harboring human microbiota (known as human-microbiota-associated mice). With the same genetic system and animal model, we here investigate for the first time the ability of L. casei to reinitiate synthesis in the different digestive tract compartments. In this study, most ingested L. casei cells transited from the stomach to the duodenum-jejunum within 1 h postingestion. No luciferase activity was observed in these digestive tract compartments after the first hour. At later times, the bulk of bacteria had transited to the ileum and the cecum. Luciferase synthesis was detected between 1.5 and 2.0 h postingestion at the ileal level and from 1.5 h to at least 6.0 h postingestion in the cecum, where the activity remained at a maximum level. These results demonstrate that ingested L. casei (derivative of the DN-114 001 strain) administered via a fermented milk has already reinitiated protein synthesis when it reaches the ileal and cecal compartments.

  7. A mouse model of pulmonary metastasis from spontaneous osteosarcoma monitored in vivo by Luciferase imaging.

    Directory of Open Access Journals (Sweden)

    Silvia Miretti

    Full Text Available BACKGROUND: Osteosarcoma (OSA is lethal when metastatic after chemotherapy and/or surgical treatment. Thus animal models are necessary to study the OSA metastatic spread and to validate novel therapies able to control the systemic disease. We report the development of a syngeneic (Balb/c murine OSA model, using a cell line derived from a spontaneous murine tumor. METHODOLOGY: The tumorigenic and metastatic ability of OSA cell lines were assayed after orthotopic injection in mice distal femur. Expression profiling was carried out to characterize the parental and metastatic cell lines. Cells from metastases were propagated and engineered to express Luciferase, in order to follow metastases in vivo. PRINCIPAL FINDINGS: Luciferase bioluminescence allowed to monitor the primary tumor growth and revealed the appearance of spontaneous pulmonary metastases. In vivo assays showed that metastasis is a stable property of metastatic OSA cell lines after both propagation in culture and luciferase trasduction. When compared to parental cell line, both unmodified and genetically marked metastatic cells, showed comparable and stable differential expression of the enpp4, pfn2 and prkcd genes, already associated to the metastatic phenotype in human cancer. CONCLUSIONS: This OSA animal model faithfully recapitulates some of the most important features of the human malignancy, such as lung metastatization. Moreover, the non-invasive imaging allows monitoring the tumor progression in living mice. A great asset of this model is the metastatic phenotype, which is a stable property, not modifiable after genetic manipulation.

  8. Using the dioxin receptor-calux in vitro bioassay to screen marine harbor sediments for compounds with a dioxin-like mode of action

    NARCIS (Netherlands)

    Stronkhorst, J.; Leonards, P.E.G.; Murk, A.J.

    2002-01-01

    The presence of dioxin-like compounds in sediments from harbors and reference sites along the Dutch coast was investigated using the dioxin receptor–chemically activated luciferase gene expression (DR-CALUX) bioassay. The DR-CALUX response varied between 0.2 and 136 ng/kg dry weight expressed in

  9. Excited-State Dynamics of Oxyluciferin in Firefly Luciferase

    KAUST Repository

    Snellenburg, Joris J.

    2016-11-23

    The color variations of light emitted by some natural and mutant luciferases are normally attributed to collective factors referred to as microenvironment effects; however, the exact nature of these interactions between the emitting molecule (oxyluciferin) and the active site remains elusive. Although model studies of noncomplexed oxyluciferin and its variants have greatly advanced the understanding of its photochemistry, extrapolation of the conclusions to the real system requires assumptions about the polarity and proticity of the active site. To decipher the intricate excited-state dynamics, global and target analysis is performed here for the first time on the steady-state and time-resolved spectra of firefly oxyluciferin complexed with luciferase from the Japanese firefly (Luciola cruciata). The experimental steady-state and time resolved luminescence spectra of the oxyluciferin/luciferase complex in solution are compared with the broadband time-resolved firefly bioluminescence recorded in vivo. The results demonstrate that de-excitation of the luminophore results in a complex cascade of photoinduced proton transfer processes and can be interpreted by the pH dependence of the emitted light. It is confirmed that proton transfer is the central event in the spectrochemistry of this system for which any assignment of the pH dependent emission to a single chemical species would be an oversimplification.

  10. Excited-State Dynamics of Oxyluciferin in Firefly Luciferase

    KAUST Repository

    Snellenburg, Joris J.; Laptenok, Sergey P.; DeSa, Richard J.; Naumov, Pance; Solntsev, Kyril M.

    2016-01-01

    The color variations of light emitted by some natural and mutant luciferases are normally attributed to collective factors referred to as microenvironment effects; however, the exact nature of these interactions between the emitting molecule (oxyluciferin) and the active site remains elusive. Although model studies of noncomplexed oxyluciferin and its variants have greatly advanced the understanding of its photochemistry, extrapolation of the conclusions to the real system requires assumptions about the polarity and proticity of the active site. To decipher the intricate excited-state dynamics, global and target analysis is performed here for the first time on the steady-state and time-resolved spectra of firefly oxyluciferin complexed with luciferase from the Japanese firefly (Luciola cruciata). The experimental steady-state and time resolved luminescence spectra of the oxyluciferin/luciferase complex in solution are compared with the broadband time-resolved firefly bioluminescence recorded in vivo. The results demonstrate that de-excitation of the luminophore results in a complex cascade of photoinduced proton transfer processes and can be interpreted by the pH dependence of the emitted light. It is confirmed that proton transfer is the central event in the spectrochemistry of this system for which any assignment of the pH dependent emission to a single chemical species would be an oversimplification.

  11. Grays Harbor Paper

    Energy Technology Data Exchange (ETDEWEB)

    Quigg, B. [Grays Harbor Paper, Hoquiam, WA (United States)

    2009-07-01

    Wood waste biomass boilers are used at Grays Harbor Paper in Hoquiam, Washington. This presentation showed that large volumes of biomass are left after a traditional clearcut. The opportunities and challenges of collecting branches, tops and stumps from this wet coastal climate were outlined. The paper described some of the low-tech methods for picking up branches, stumps and woody debris. It included several photographs of custom logging machines for timber harvest, including a brush grapple slasher, a shearer shovel, chippers, grinders, slicesaws, trucks, trailers and caterpillar log loaders for handling slash. The slash recovery program relies on innovative harvesting machines that convert scattered logging slash into bundles that can be easily collected, transported, and stored for use in existing facilities that utilize wood fiber for fuel. figs.

  12. Application of luciferase assay for ATP to antimicrobial drug susceptibility

    Science.gov (United States)

    Chappelle, E. W.; Picciolo, G. L.; Vellend, H.; Tuttle, S. A.; Barza, M. J.; Weinstein, L. (Inventor)

    1977-01-01

    The susceptibility of bacteria, particularly those derived from body fluids, to antimicrobial agents is determined in terms of an ATP index measured by culturing a bacterium in a growth medium. The amount of ATP is assayed in a sample of the cultured bacterium by measuring the amount of luminescent light emitted when the bacterial ATP is reacted with a luciferase-luciferin mixture. The sample of the cultured bacterium is subjected to an antibiotic agent. The amount of bacterial adenosine triphosphate is assayed after treatment with the antibiotic by measuring the luminescent light resulting from the reaction. The ATP index is determined from the values obtained from the assay procedures.

  13. Alaska Harbor Seal Glacial Surveys

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Floating glacial ice serves as a haul-out substrate for a significant number (10-15%) of Alaskan harbor seals, and thus surveying tidewater glacial fjords is an...

  14. Luciferase-Specific Coelenterazine Analogues for Optical Contamination-Free Bioassays

    OpenAIRE

    Ryo Nishihara; Masahiro Abe; Shigeru Nishiyama; Daniel Citterio; Koji Suzuki; Sung Bae Kim

    2017-01-01

    Spectral overlaps among the multiple optical readouts commonly cause optical contamination in fluorescence and bioluminescence. To tackle this issue, we created five-different lineages of coelenterazine (CTZ) analogues designed to selectively illuminate a specific luciferase with unique luciferase selectivity. In the attempt, we found that CTZ analogues with ethynyl or styryl groups display dramatically biased bioluminescence to specific luciferases and pHs by modifying the functional groups ...

  15. Development of microLIPS (Luciferase Immunoprecipitation Systems): a novel microfluidic assay for rapid serum antibody detection

    Science.gov (United States)

    Chandrangsu, Matt; Burbelo, Peter D.; Iadarola, Michael J.; Smith, Paul D.; Morgan, Nicole Y.

    2012-06-01

    There is considerable interest in the development of rapid, point-of-care antibody detection for the diagnosis of infectious and auto-immune diseases. In this paper, we present work on the development of a self-contained microfluidic format for the Luciferase Immunoprecipitation Systems (LIPS) assay. Whereas the majority of immunoassays for antigen-specific antibodies employ either bacteria- or yeast-expressed proteins and require the use of secondary antibodies, the LIPS technique uses a fusion protein comprised of a Renilla luciferase reporter and the antigen of interest produced via mammalian cell culture, ensuring the addition of mammalian post-translational modifications. Patient serum is mixed with the fusion protein and passed over immobilized Protein A/G; after washing, the only remaining luciferase-tagged antigens are those retained by specific antibodies. These can be quantitatively measured using chemiluminescence upon the introduction of coelenterazine. The assay has been successfully employed for a wide variety of diseases in a microwell format. We report on a recent demonstration of rapid HSV-2 diagnosis with the LIPS assay in a microfluidic format, using one microliter of serum and obtaining results in under ten minutes. We will also discuss recent progress on two fronts, both aimed at the deployment of this technology in the field: first, simplifying assay operation through the automation of flow control using power-free means; and second, efforts to increase signal levels, primarily through strategies to increase antibody binding capacity, in order to move towards portable battery powered electronics.

  16. Old Harbor Scammon Bay Hydro Feasibility

    Energy Technology Data Exchange (ETDEWEB)

    Brent Petrie

    2007-06-27

    The grantee, Alaska Village Electric Cooperative (AVEC), is a non-profit member owned rural electric generation and distribution cooperative. The proposed Project is located near the community of Old Harbor, Alaska. Old Harbor is on the southeastern coast of Kodiak Island, approximately 70 miles southwest of the City of Kodiak and 320 miles southwest of Anchorage. In 1998 sufficient information had been developed to apply for a license to construct the project and the cost was estimated to be $2,445,000 for a 500 KW project on Lagoon Creek. Major features of the project included an eight-foot high diversion dam on Mountain Creek, a desander box, a 9,800-foot long penstock to the powerhouse on Lagoon Creek, and a 5,500-foot long access road. It was also anticipated that the project could provide an additional source of water to Old Harbor. The report details the history and lessons learned in designing and permiting the proposed hydroelectric facility.

  17. Geoscience rediscovers Phoenicia's buried harbors

    Science.gov (United States)

    Marriner, Nick; Morhange, Christophe; Doumet-Serhal, Claude; Carbonel, Pierre

    2006-01-01

    After centuries of archaeological debate, the harbors of Phoenicia's two most important city states, Tyre and Sidon, have been rediscovered, and including new geoarcheological results reveal how, where, and when they evolved after their Bronze Age foundations. The early ports lie beneath their present urban centers, and we have indentified four harbor phases. (1) During the Bronze Age, Tyre and Sidon were characterized by semi-open marine coves that served as protoharbors. (2) Biostratigraphic and lithostratigraphic data indicate the presence of early artificial basins after the first millennium B.C. (3) The harbors reached their apogees during the Greco-Roman and Byzantine periods. (4) Silting up and coastal progradation led to burial of the medieval basins, lost until now.

  18. Luciferase-Zinc-Finger System for the Rapid Detection of Pathogenic Bacteria.

    Science.gov (United States)

    Shi, Chu; Xu, Qing; Ge, Yue; Jiang, Ling; Huang, He

    2017-08-09

    Rapid and reliable detection of pathogenic bacteria is crucial for food safety control. Here, we present a novel luciferase-zinc finger system for the detection of pathogens that offers rapid and specific profiling. The system, which uses a zinc-finger protein domain to probe zinc finger recognition sites, was designed to bind the amplified conserved regions of 16S rDNA, and the obtained products were detected using a modified luciferase. The luciferase-zinc finger system not only maintained luciferase activity but also allowed the specific detection of different bacterial species, with a sensitivity as low as 10 copies and a linear range from 10 to 10 4 copies per microliter of the specific PCR product. Moreover, the system is robust and rapid, enabling the simultaneous detection of 6 species of bacteria in artificially contaminated samples with excellent accuracy. Thus, we envision that our luciferase-zinc finger system will have far-reaching applications.

  19. Satellite Monitoring of Boston Harbor Water Quality: Initial Investigations

    Science.gov (United States)

    Sheldon, P.; Chen, R. F.; Schaaf, C.; Pahlevan, N.; Lee, Z.

    2016-02-01

    The transformation of Boston Harbor from the "dirtiest in America" to a National Park Area is one of the most remarkable estuarine recoveries in the world. A long-term water quality dataset from 1991 to present exists in Boston Harbor due to a $3. 8 billion lawsuit requiring the harbor clean-up. This project uses discrete water sampling and underway transects with a towed vehicle coordinated with Landsat 7 and Landsat 8 to create surface maps of chlorophyll a (Chl a), dissolved organic matter (CDOM and DOC), total suspended solids (TSS), diffuse attenuation coefficient (Kd_490), and photic depth in Boston Harbor. In addition, 3 buoys have been designed, constructed, and deployed in Boston Harbor that measure Chl a and CDOM fluorescence, optical backscatter, salinity, temperature, and meteorological parameters. We are initially using summer and fall of 2015 to develop atmospheric corrections for conditions in Boston Harbor and develop algorithms for Landsat 8 data to estimate in water photic depth, TSS, Chl a, Kd_490, and CDOM. We will report on initial buoy and cruise data and show 2015 Landsat-derived distributions of water quality parameters. It is our hope that once algorithms for present Landsat imagery can be developed, historical maps of water quality can be constructed using in water data back to 1991.

  20. 77 FR 19967 - Safety Zone, Port of Dutch Harbor; Dutch Harbor, AK

    Science.gov (United States)

    2012-04-03

    ...-AA00 Safety Zone, Port of Dutch Harbor; Dutch Harbor, AK AGENCY: Coast Guard, DHS. ACTION: Notice of proposed rulemaking. SUMMARY: The Coast Guard proposes temporary safety zones in the Port of Dutch Harbor... Dutch Harbor, Alaska, and the adjacent territorial sea due to additional vessel traffic associated with...

  1. Army Engineers at Pearl Harbor

    Science.gov (United States)

    2011-01-01

    siblings, he was the grandson of David Belden Lyman—a Christian missionary from New England who settled in the Hilo , Hawaii area—and the descendent of...of Hawaii appeared over Oahu. Some headed for Ameri- can warships at Pearl Harbor and the planes on the ground at nearby Hickam Field; oth- ers...hit Schofield Barracks, Wheeler Field, and Bellows Field. USACE in Hawaii con- sisted of Soldier-engineers in the Army’s Hawaiian Depart- ment and

  2. Adenovirus-assisted lipofection: efficient in vitro gene transfer of luciferase and cytosine deaminase to human smooth muscle cells.

    Science.gov (United States)

    Kreuzer, J; Denger, S; Reifers, F; Beisel, C; Haack, K; Gebert, J; Kübler, W

    1996-07-01

    Smooth muscle cells (SMC) are a central cell type involved in multiple processes of coronary artery diseases including restenosis and therefore are major target cells for different aspects of gene transfer. Previous attempts to transfect primary arterial cells using different techniques like liposomes, CaPO4 and electroporation resulted in only low transfection efficiency. The development of recombinant adenoviruses dramatically improved the delivery of foreign genes into different cell types including SMC. However, cloning and identification of recombinants remain difficult and time-consuming techniques. The present study demonstrates that a complex consisting of reporter plasmid encoding firefly luciferase (pLUC), polycationic liposomes and replication-deficient adenovirus was able to yield very high in vitro transfection of primary human smooth muscle cells under optimized conditions. The technique of adenovirus-assisted lipofection (AAL) increases transfer and expression of plasmid DNA in human smooth muscle cells in vitro up to 1000-fold compared to lipofection. To verify the applicability of AAL for gene transfer into human smooth muscle cells we studied a gene therapy approach to suppress proliferation of SMC in vitro, using the prokaryotic cytosine deaminase gene (CD) which enables transfected mammalian cells to deaminate 5-fluorocytosine (5-FC) to the highly toxic 5-fluorouracil (5-FU). The effect of a transient CD expression on RNA synthesis was investigated by means of a cotransfection with a RSV-CD expression plasmid and the luciferase reporter plasmid. Western blot analysis demonstrated high expression of CD protein in transfected SMC. Cotransfected SMC demonstrated two-fold less luciferase activity in the presence of 5-FC (5 mmol/l) after 48 h compared to cells transfected with a non-CD coding plasmid. The data demonstrate that a transient expression of CD could be sufficient to reduce the capacity of protein synthesis in human SMC. This simple and

  3. Luciferase-Specific Coelenterazine Analogues for Optical Contamination-Free Bioassays.

    Science.gov (United States)

    Nishihara, Ryo; Abe, Masahiro; Nishiyama, Shigeru; Citterio, Daniel; Suzuki, Koji; Kim, Sung Bae

    2017-04-19

    Spectral overlaps among the multiple optical readouts commonly cause optical contamination in fluorescence and bioluminescence. To tackle this issue, we created five-different lineages of coelenterazine (CTZ) analogues designed to selectively illuminate a specific luciferase with unique luciferase selectivity. In the attempt, we found that CTZ analogues with ethynyl or styryl groups display dramatically biased bioluminescence to specific luciferases and pHs by modifying the functional groups at the C-2 and C-6 positions of the imidazopyradinone backbone of CTZ. The optical contamination-free feature was exemplified with the luciferase-specific CTZ analogues, which illuminated anti-estrogenic and rapamycin activities in a mixture of optical probes. This unique bioluminescence platform has great potential for specific and high throughput imaging of multiple optical readouts in bioassays without optical contamination.

  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. Giant Cell Arteritis of the Female Genital Tract With Occult Temporal Arteritis and Marginal Zone Lymphoma Harboring Novel 20q Deletion: A Case Report and Literature Review.

    Science.gov (United States)

    Pradhan, Dinesh; Amin, Rajnikant M; Jones, Miroslawa W; Surti, Urvashi; Parwani, Anil V

    2016-02-01

    Giant cell arteritis (GCA) is an immunologically mediated vasculitis of large and medium-sized vessels, typically affecting the cranial arteries and usually occurring in the elderly. GCA of the female genital tract is extremely rare with only 31 cases reported in the English literature. An 83-year-old white female with postmenopausal vaginal bleeding revealed an endometrial polyp on pelvic ultrasonography following which polypectomy and subsequently hysterectomy with bilateral salpingo-oophorectomy was done. Microscopy revealed a well-differentiated endometrioid adenocarcinoma. Interestingly, classic GCA involving numerous small to medium-sized arteries of the cervix, myometrium, bilateral fallopian tubes, and ovaries was also identified. Hematologic evaluation revealed marginal zone lymphoma with an exceptionally rare 20q deletion. Bilateral temporal artery biopsy was done subsequently, which exhibited GCA on microscopy. Corticosteroid was started that improved her polymyalgia rheumatica symptoms. The patient is on follow-up for 3 years and is doing well. To our knowledge, this is the first case of GCA of the female genital tract associated with a lymphoma and the second case of marginal zone lymphoma with the novel 20q deletion. © The Author(s) 2015.

  6. Characterization of CG6178 gene product with high sequence similarity to firefly luciferase in Drosophila melanogaster.

    Science.gov (United States)

    Oba, Yuichi; Ojika, Makoto; Inouye, Satoshi

    2004-03-31

    This is the first identification of a long-chain fatty acyl-CoA synthetase in Drosophila by enzymatic characterization. The gene product of CG6178 (CG6178) in Drosophila melanogaster genome, which has a high sequence similarity to firefly luciferase, has been expressed and characterized. CG6178 showed long-chain fatty acyl-CoA synthetic activity in the presence of ATP, CoA and Mg(2+), suggesting a fatty acyl adenylate is an intermediate. Recently, it was revealed that firefly luciferase has two catalytic functions, monooxygenase (luciferase) and AMP-mediated CoA ligase (fatty acyl-CoA synthetase). However, unlike firefly luciferase, CG6178 did not show luminescence activity in the presence of firefly luciferin, ATP, CoA and Mg(2+). The enzymatic properties of CG6178 including substrate specificity, pH dependency and optimal temperature were close to those of firefly luciferase and rat fatty acyl-CoA synthetase. Further, phylogenic analyses strongly suggest that the firefly luciferase gene may have evolved from a fatty acyl-CoA synthetase gene as a common ancestral gene.

  7. Cloning and characterization of luciferase from a Fijian luminous click beetle.

    Science.gov (United States)

    Mitani, Yasuo; Futahashi, Ryo; Niwa, Kazuki; Ohba, Nobuyoshi; Ohmiya, Yoshihiro

    2013-01-01

    Luminous click beetle is distributed almost exclusively in Central and South America with a single genus in Melanesia. Among these click beetles, the description of Melanesian species has been fragmentary, and its luciferase gene and phylogenetic relation to other click beetles still remain uncertain. We collected a living luminous click beetle, Photophorus jansonii in Fiji. It emits green-yellow light from two spots on the pronotum and has no ventral luminous organ. Here, we cloned a luciferase gene from this insect by RT-PCR. The deduced amino acid sequence showed high identity of ~85% to the luciferases derived from other click beetle species. The luciferase of the Fijian click beetle was produced as a recombinant protein to characterize its biochemical properties. The Km for D-luciferin and ATP were 173 and 270 μm, respectively. The luciferase was pH-insensitive and the spectrum measured at pH 8.0 showed a peak at 559 nm, which was in the range of green-yellow light as seen in the luminous spot of the living Fijian click beetle. The Fijian click beetle luciferase was assigned to the Elateridae clade by a phylogenetic analysis, but it made a clearly different branch from Pyrophorus group examined in this study. © 2013 The American Society of Photobiology.

  8. 33 CFR 125.15 - Access to waterfront facilities, and port and harbor areas, including vessels and harbor craft...

    Science.gov (United States)

    2010-07-01

    ..., and port and harbor areas, including vessels and harbor craft therein. 125.15 Section 125.15....15 Access to waterfront facilities, and port and harbor areas, including vessels and harbor craft....09 to those waterfront facilities, and port and harbor areas, including vessels and harbor craft...

  9. A dual-color luciferase assay system reveals circadian resetting of cultured fibroblasts by co-cultured adrenal glands.

    Directory of Open Access Journals (Sweden)

    Takako Noguchi

    Full Text Available In mammals, circadian rhythms of various organs and tissues are synchronized by pacemaker neurons in the suprachiasmatic nucleus (SCN of the hypothalamus. Glucocorticoids released from the adrenal glands can synchronize circadian rhythms in other tissues. Many hormones show circadian rhythms in their plasma concentrations; however, whether organs outside the SCN can serve as master synchronizers to entrain circadian rhythms in target tissues is not well understood. To further delineate the function of the adrenal glands and the interactions of circadian rhythms in putative master synchronizing organs and their target tissues, here we report a simple co-culture system using a dual-color luciferase assay to monitor circadian rhythms separately in various explanted tissues and fibroblasts. In this system, circadian rhythms of organs and target cells were simultaneously tracked by the green-emitting beetle luciferase from Pyrearinus termitilluminans (ELuc and the red-emitting beetle luciferase from Phrixothrix hirtus (SLR, respectively. We obtained tissues from the adrenal glands, thyroid glands, and lungs of transgenic mice that expressed ELuc under control of the promoter from a canonical clock gene, mBmal1. The tissues were co-cultured with Rat-1 fibroblasts as representative target cells expressing SLR under control of the mBmal1 promoter. Amplitudes of the circadian rhythms of Rat-1 fibroblasts were potentiated when the fibroblasts were co-cultured with adrenal gland tissue, but not when co-cultured with thyroid gland or lung tissue. The phases of Rat-1 fibroblasts were reset by application of adrenal gland tissue, whereas the phases of adrenal gland tissue were not influenced by Rat-1 fibroblasts. Furthermore, the effect of the adrenal gland tissue on the fibroblasts was blocked by application of a glucocorticoid receptor (GR antagonist. These results demonstrate that glucocorticoids are strong circadian synchronizers for fibroblasts and that

  10. Construction and characterization of a recombinant yellow fever virus stably expressing Gaussia luciferase

    Directory of Open Access Journals (Sweden)

    TELISSA C. KASSAR

    Full Text Available ABSTRACT Yellow fever is an arthropod-borne viral disease that still poses high public health concerns, despite the availability of an effective vaccine. The development of recombinant viruses is of utmost importance for several types of studies, such as those aimed to dissect virus-host interactions and to search for novel antiviral strategies. Moreover, recombinant viruses expressing reporter genes may greatly facilitate these studies. Here, we report the construction of a recombinant yellow fever virus (YFV expressing Gaussia luciferase (GLuc (YFV-GLuc. We show, through RT-PCR, sequencing and measurement of GLuc activity, that stability of the heterologous gene was maintained after six passages. Furthermore, a direct association between GLuc expression and viral replication was observed (r2=0.9967, indicating that measurement of GLuc activity may be used to assess viral replication in different applications. In addition, we evaluated the use of the recombinant virus in an antiviral assay with recombinant human alfa-2b interferon. A 60% inhibition of GLuc expression was observed in cells infected with YFV-GLuc and incubated with IFN alfa-2b. Previously tested on YFV inhibition by plaque assays indicated a similar fold-decrease in viral replication. These results are valuable as they show the stability of YFV-GLuc and one of several possible applications of this construct.

  11. CRSMP Potential Harbor Borrow Sites 2012

    Data.gov (United States)

    California Natural Resource Agency — Harbor locations as identified originally in the California Shoreline Database compiled by Noble Consultants (Jon Moore) for California Department of Boating and...

  12. Crystal structure of the bacterial luciferase/flavin complex provides insight into the function of the beta subunit.

    Science.gov (United States)

    Campbell, Zachary T; Weichsel, Andrzej; Montfort, William R; Baldwin, Thomas O

    2009-07-07

    Bacterial luciferase from Vibrio harveyi is a heterodimer composed of a catalytic alpha subunit and a homologous but noncatalytic beta subunit. Despite decades of enzymological investigation, structural evidence defining the active center has been elusive. We report here the crystal structure of V. harveyi luciferase bound to flavin mononucleotide (FMN) at 2.3 A. The isoalloxazine ring is coordinated by an unusual cis-Ala-Ala peptide bond. The reactive sulfhydryl group of Cys106 projects toward position C-4a, the site of flavin oxygenation. This structure also provides the first data specifying the conformations of a mobile loop that is crystallographically disordered in both prior crystal structures [(1995) Biochemistry 34, 6581-6586; (1996) J. Biol. Chem. 271, 21956 21968]. This loop appears to be a boundary between solvent and the active center. Within this portion of the protein, a single contact was observed between Phe272 of the alpha subunit, not seen in the previous structures, and Tyr151 of the beta subunit. Substitutions at position 151 on the beta subunit caused reductions in activity and total quantum yield. Several of these mutants were found to have decreased affinity for reduced flavin mononucleotide (FMNH(2)). These findings partially address the long-standing question of how the beta subunit stabilizes the active conformation of the alpha subunit, thereby participating in the catalytic mechanism.

  13. 33 CFR 100.109 - Winter Harbor Lobster Boat Race, Winter Harbor, ME.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Winter Harbor Lobster Boat Race, Winter Harbor, ME. 100.109 Section 100.109 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY REGATTAS AND MARINE PARADES SAFETY OF LIFE ON NAVIGABLE WATERS § 100.109 Winter Harbor...

  14. The Suez Canal and the petroleum harbors

    International Nuclear Information System (INIS)

    Anon.

    1997-01-01

    The Suez Canal is the second longest channel in the world and allows to save 60% of the travel time between the petroleum harbors of the Arabic peninsula and Europe. This short paper gives a summary of the main petroleum harbors activity along the channel from the Red sea to the Mediterranean sea. (J.S.)

  15. Frequency Domain Response at Pacific Coast Harbors to Major Tsunamis of 2005-2011

    Science.gov (United States)

    Xing, Xiuying; Kou, Zhiqing; Huang, Ziyi; Lee, Jiin-Jen

    2013-06-01

    Tsunamis waves caused by submarine earthquake or landslide might contain large wave energy, which could cause significant human loss and property damage locally as well as in distant region. The response of three harbors located at the Pacific coast (i.e. Crescent City Harbor, Los Angeles/Long Beach Port, and San Diego Harbor) to six well-known tsunamis events generated (both near-field and far-field) between 2005 and 2011 are examined and simulated using a hybrid finite element numerical model in frequency domain. The model incorporated the effects of wave refraction, wave diffraction, partial wave reflection from boundaries, entrance and bottom energy dissipation. It can be applied to harbor regions with arbitrary shapes and variable water depth. The computed resonant periods or modes of oscillation for three harbors are in good agreement with the energy spectral analysis of the time series of water surface elevations recorded at tide gauge stations inside three harbors during the six tsunamis events. The computed wave induced currents based on the present model are also in qualitative agreement with some of the reported eye-witness accounts absence of reliable current data. The simulated results show that each harbor responded differently and significantly amplified certain wave period(s) of incident wave trains according to the shape, topography, characteristic dimensions and water depth of the harbor basins.

  16. DEPOT MAINTENANCE: Key Financial Issues for Consolidations at Pearl Harbor and Elsewhere Are Still Unresolved

    National Research Council Canada - National Science Library

    2001-01-01

    .... In September 1999, we reported that the preliminary results of the ongoing Pearl Harbor pilot were mixed and recommended that the Departments of Defense and the Navy address unresolved issues related...

  17. Human CRF2 α and β splice variants: pharmacological characterization using radioligand binding and a luciferase gene expression assay

    International Nuclear Information System (INIS)

    Ardati, A.; Goetschy, V.; Gottowick, J.; Henriot, S.; Deuschle, U.; Kilpatrick, G.J.; Valdenaire, O.

    1999-01-01

    Corticotropin releasing factor (CRF) receptors belong to the super-family of G protein-coupled receptors. These receptors are classified into two subtypes (CRF 1 and CRF 2 ). Both receptors are positively coupled to adenylyl cyclase but they have a distinct pharmacology and distribution in brain. Two isoforms belonging to the CRF 2 subtype receptors, CRF 2α and CRF 2β , have been identified in rat and man. The neuropeptides CRF and urocortin mediate their actions through this CRF G protein-coupled receptor family. In this report, we describe the pharmacological characterization of the recently identified hCRF 2β receptor. We have used radioligand binding with [ 125 I]-tyr 0 -sauvagine and a gene expression assay in which the firefly luciferase gene expression is under the control of cAMP responsive elements. Association kinetics of [ 125 I]-tyr 0 -sauvagine binding to the hCRF 2β receptor were monophasic while dissociation kinetics were biphasic, in agreement with the kinetics results obtained with the hCRF 2α receptor. Saturation binding analysis revealed two affinity states in HEK 293 cells with binding parameters in accord with those determined kinetically and with parameters obtained with the hCRF 2α receptor. A non-hydrolysable GTP analog, Gpp(NH)p, reduced the high affinity binding of [ 125 I]-tyr 0 -sauvagine to both hCRF 2 receptor isoforms in a similar manner. The rank order of potency of CRF agonist peptides in competition experiments was identical for both hCRF 2 α-helical CRF (9-41) oCRF). Similarly, agonist potency was similar for the two isoforms when studied using the luciferase gene reporter system. The peptide antagonist α-helical CRF (9-41) exhibited a non-competitive antagonism of urocortin-stimulated luciferase expression with both hCRF 2 receptor isoforms. Taken together, these results indicate that the pharmacological profiles of the CRF 2 splice variants are identical. This indicates that the region of the N-terminus that varies

  18. Plasmodium falciparum transfected with ultra bright NanoLuc luciferase offers high sensitivity detection for the screening of growth and cellular trafficking inhibitors.

    Directory of Open Access Journals (Sweden)

    Mauro F Azevedo

    Full Text Available Drug discovery is a key part of malaria control and eradication strategies, and could benefit from sensitive and affordable assays to quantify parasite growth and to help identify the targets of potential anti-malarial compounds. Bioluminescence, achieved through expression of exogenous luciferases, is a powerful tool that has been applied in studies of several aspects of parasite biology and high throughput growth assays. We have expressed the new reporter NanoLuc (Nluc luciferase in Plasmodium falciparum and showed it is at least 100 times brighter than the commonly used firefly luciferase. Nluc brightness was explored as a means to achieve a growth assay with higher sensitivity and lower cost. In addition we attempted to develop other screening assays that may help interrogate libraries of inhibitory compounds for their mechanism of action. To this end parasites were engineered to express Nluc in the cytoplasm, the parasitophorous vacuole that surrounds the intraerythrocytic parasite or exported to the red blood cell cytosol. As proof-of-concept, these parasites were used to develop functional screening assays for quantifying the effects of Brefeldin A, an inhibitor of protein secretion, and Furosemide, an inhibitor of new permeation pathways used by parasites to acquire plasma nutrients.

  19. Environmental Assessment for Boston Harbor Maintenance Dredging, Boston, Massachusetts.

    Science.gov (United States)

    1981-12-01

    Harbor was developed by Jerome et al (1966), Chesmore et al (1971) and Iwanowicz et al. (1973). The studies on the Lower Mystic River were concentrated in... Iwanowicz et al. (1973) and this data should be referred to for detailed information. Waters overlying the shellfish beds are contaminated by wastes...DMRP Technical Report DS-78-5, Environmental Laboratory, U.S. Army Engineer Waterways Experiment Station, Vicksburg, Mississippi. Iwanowicz , H. R., R D

  20. Genetics Home Reference: Floating-Harbor syndrome

    Science.gov (United States)

    ... Patton MA, Hurst J, Donnai D, McKeown CM, Cole T, Goodship J. Floating-Harbor syndrome. J Med ... medicine? What is newborn screening? New Pages Lyme disease Fibromyalgia White-Sutton syndrome All New & Updated Pages ...

  1. F-18 Labeled Diabody-Luciferase Fusion Proteins for Optical-ImmunoPET

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Anna M. [Univ. of California, Los Angeles, CA (United States)

    2013-01-18

    The goal of the proposed work is to develop novel dual-labeled molecular imaging probes for multimodality imaging. Based on small, engineered antibodies called diabodies, these probes will be radioactively tagged with Fluorine-18 for PET imaging, and fused to luciferases for optical (bioluminescence) detection. Performance will be evaluated and validated using a prototype integrated optical-PET imaging system, OPET. Multimodality probes for optical-PET imaging will be based on diabodies that are dually labeled with 18F for PET detection and fused to luciferases for optical imaging. 1) Two sets of fusion proteins will be built, targeting the cell surface markers CEA or HER2. Coelenterazine-based luciferases and variant forms will be evaluated in combination with native substrate and analogs, in order to obtain two distinct probes recognizing different targets with different spectral signatures. 2) Diabody-luciferase fusion proteins will be labeled with 18F using amine reactive [18F]-SFB produced using a novel microwave-assisted, one-pot method. 3) Sitespecific, chemoselective radiolabeling methods will be devised, to reduce the chance that radiolabeling will inactivate either the target-binding properties or the bioluminescence properties of the diabody-luciferase fusion proteins. 4) Combined optical and PET imaging of these dual modality probes will be evaluated and validated in vitro and in vivo using a prototype integrated optical-PET imaging system, OPET. Each imaging modality has its strengths and weaknesses. Development and use of dual modality probes allows optical imaging to benefit from the localization and quantitation offered by the PET mode, and enhances the PET imaging by enabling simultaneous detection of more than one probe.

  2. The cAMP-dependent protein kinase inhibitor H-89 attenuates the bioluminescence signal produced by Renilla Luciferase.

    Directory of Open Access Journals (Sweden)

    Katie J Herbst

    2009-05-01

    Full Text Available Investigations into the regulation and functional roles of kinases such as cAMP-dependent protein kinase (PKA increasingly rely on cellular assays. Currently, there are a number of bioluminescence-based assays, for example reporter gene assays, that allow the study of the regulation, activity, and functional effects of PKA in the cellular context. Additionally there are continuing efforts to engineer improved biosensors that are capable of detecting real-time PKA signaling dynamics in cells. These cell-based assays are often utilized to test the involvement of PKA-dependent processes by using H-89, a reversible competitive inhibitor of PKA.We present here data to show that H-89, in addition to being a competitive PKA inhibitor, attenuates the bioluminescence signal produced by Renilla luciferase (RLuc variants in a population of cells and also in single cells. Using 10 microM of luciferase substrate and 10 microM H-89, we observed that the signal from RLuc and RLuc8, an eight-point mutation variant of RLuc, in cells was reduced to 50% (+/-15% and 54% (+/-14% of controls exposed to the vehicle alone, respectively. In vitro, we showed that H-89 decreased the RLuc8 bioluminescence signal but did not compete with coelenterazine-h for the RLuc8 active site, and also did not affect the activity of Firefly luciferase. By contrast, another competitive inhibitor of PKA, KT5720, did not affect the activity of RLuc8.The identification and characterization of the adverse effect of H-89 on RLuc signal will help deconvolute data previously generated from RLuc-based assays looking at the functional effects of PKA signaling. In addition, for the current application and future development of bioluminscence assays, KT5720 is identified as a more suitable PKA inhibitor to be used in conjunction with RLuc-based assays. These principal findings also provide an important lesson to fully consider all of the potential effects of experimental conditions on a cell

  3. Madaket Harbor, Nantucket, Massachusetts. Water Resources Improvement.

    Science.gov (United States)

    1977-07-01

    will continue to be, important increases in the recreational use of land and water. The harbor area is an important arena for commercial shellfishing...an important arena for commercial shell fishing. The past few years have seen a rather rapid increase in residential land use. Construction has...beamc. Tnis material will be re-deposited,, viaj troio it 1-apfro1inr ox prior location. j, MADAKET HARBOR NANTUCKET, MASSACHUSETTS FEASIBILITY

  4. Emergence of colistin-resistant Escherichia coli clinical isolates harboring mcr-1 in Vietnam.

    Science.gov (United States)

    Tada, Tatsuya; Nhung, Pham Hong; Shimada, Kayo; Tsuchiya, Mitsuhiro; Phuong, Doan Mai; Anh, Nguyen Quoc; Ohmagari, Norio; Kirikae, Teruo

    2017-10-01

    The mcr-1 was first detected on a plasmid in colistin-resistant Escherichia coli from livestock and patients in China. We described here the emergence of colistin-resistant E. coli clinical isolates harboring mcr-1 on the chromosomes in Vietnam. To our knowledge, this is the first report of hospital-acquired E. coli isolates harboring mcr-1 in a medical setting in Vietnam. Copyright © 2017. Published by Elsevier Ltd.

  5. Charleston Harbor, SC, Regional Sediment Management Study: Beneficial Use of Dredged Material through Nearshore Placement

    Science.gov (United States)

    2017-05-01

    Modeling System (CMS) that were developed and applied previously for the Charleston Harbor numerical modeling study (Kashlan 2013) were used in...Particle Tracking Model; Report 1: Model theory, implementation, and example applications. ERDC/CHL TR-6-20. Vicksburg, MS: U.S. Army Engineer Research ...ER D C/ CH L TR -1 7- 7 Regional Sediment Management (RSM) Program Charleston Harbor, SC, Regional Sediment Management Study

  6. Charleston Harbor, SC, Regional Sediment Management Study; Beneficial Use of Dredged Material through Nearshore Placement

    Science.gov (United States)

    2017-05-01

    Modeling System (CMS) that were developed and applied previously for the Charleston Harbor numerical modeling study (Kashlan 2013) were used in...Particle Tracking Model; Report 1: Model theory, implementation, and example applications. ERDC/CHL TR-6-20. Vicksburg, MS: U.S. Army Engineer Research ...ER D C/ CH L TR -1 7- 7 Regional Sediment Management (RSM) Program Charleston Harbor, SC, Regional Sediment Management Study

  7. 76 FR 8653 - Drawbridge Operation Regulation; Gulf Intracoastal Waterway, New Orleans Harbor, Inner Harbor...

    Science.gov (United States)

    2011-02-15

    ... Operation Regulation; Gulf Intracoastal Waterway, New Orleans Harbor, Inner Harbor Navigation Canal, New Orleans, Orleans Parish, LA AGENCY: Coast Guard, DHS. ACTION: Notice of temporary deviation from... Lock), at New Orleans, Orleans Parish, Louisiana. This deviation is necessary to replace all of the...

  8. 75 FR 78601 - Drawbridge Operation Regulation; Gulf Intracoastal Waterway, New Orleans Harbor, Inner Harbor...

    Science.gov (United States)

    2010-12-16

    ... Operation Regulation; Gulf Intracoastal Waterway, New Orleans Harbor, Inner Harbor Navigation Canal, New Orleans, Orleans Parish, LA AGENCY: Coast Guard, DHS. ACTION: Notice of temporary deviation from... Harvey Lock), at New Orleans, Orleans Parish, Louisiana. This deviation is necessary to adjust the...

  9. Step-wise addition of disulfide bridge in firefly luciferase controls color shift through a flexible loop: a thermodynamic perspective.

    Science.gov (United States)

    Nazari, Mahboobeh; Hosseinkhani, Saman; Hassani, Leila

    2013-02-01

    Multi-color bioluminescence is developed using the introduction of single/double disulfide bridges in firefly luciferase. The bioluminescence reaction, which uses luciferin, Mg(2+)-ATP and molecular oxygen to yield an electronically excited oxyluciferin, is carried out by the luciferase and emits visible light. The bioluminescence color of firefly luciferases is determined by the luciferase sequence and assay conditions. It has been proposed that the stability of a protein may increase through the introduction of a disulfide bridge that decreases the configurational entropy of unfolding. Single and double disulfide bridges are introduced into Photinus pyralis firefly luciferase to make separate mutant enzymes with a single/double bridge (C(81)-A(105)C, L(306)C-L(309)C, P(451)C-V(469)C; C(81)-A(105)C/P(451)C-V(469)C, and A(296)C-A(326)C/P(451)C-V(469)C). By introduction of disulfide bridges using site-directed mutagenesis in Photinus pyralis luciferase the color of emitted light was changed to red or kept in different extents. The bioluminescence color shift occurred with displacement of a critical loop in the luciferase structure without any change in green emitter mutants. Thermodynamic analysis revealed that among mutants, L(306)C-L(309)C shows a remarkable stability against urea denaturation and also a considerable increase in kinetic stability and a clear shift in bioluminescence spectra towards red.

  10. New Harbor in Kangerlussuaq, Western Greenland

    DEFF Research Database (Denmark)

    Stenstad, Jaran Gjerlandj; Eppeland, Kjetil Grødal; Ingeman-Nielsen, Thomas

    2015-01-01

    transported by rivers from the inland ice to the inner parts of the fjord. These sediment layers reduce the water depth and prevent container- and cruiseships to dock, imposing large additional maintenance costs, and inefficient operability. Through engineering geological field and lab investigations......, a possible new harbor location around 10 km further out the fjord near Hancock Pynt, has been investigated. The onshore area was found to be highly suitable for a harbor support area, where a sub-base thickness of 1.8 m with gravel cover-layer was found adequate for the calculated design loads. Existing...

  11. a positive control plasmid for reporter gene assay

    African Journals Online (AJOL)

    STORAGESEVER

    2008-07-04

    Jul 4, 2008 ... qualification as a positive control for luciferase reporter gene assays. Key words: Reporter gene plasmid, luciferase assay, cytomegalovirus promoter/enhancer, human melanoma cell line. INTRODUCTION. Reporter genes, often called reporters, have become a precious tool in studies of gene expression ...

  12. Summary of oceanographic and water–quality measurements in West Falmouth Harbor and Buzzards Bay, Massachusetts, 2009–2010

    Science.gov (United States)

    Ganju, Neil K.; Dickhudt, Patrick J.; Thomas, Jennifer A.; Borden, Jonathan; Sherwood, Christopher R.; Montgomery, Ellyn T.; Twomey, Erin R.; Martini, Marinna A.

    2011-01-01

    This data report presents oceanographic and water-quality observations made at six locations in West Falmouth Harbor and Buzzards Bay, Massachusetts, from August 2009 to September 2010. Both Buzzards Bay and West Falmouth Harbor are estuarine embayments; the input of freshwater on the eastern margin of Buzzards Bay adjacent to Cape Cod and West Falmouth Harbor is largely due to groundwater. In West Falmouth Harbor, the groundwater that seeps into the harbor is characterized by relatively high levels of nitrate. This high nitrate load has modified the ecology of the harbor (Howes and others, 2006) and may be a significant source of nitrate to Buzzards Bay during seasons with low biological nitrate uptake. The U.S. Geological Survey undertook these measurements to improve understanding of circulation, residence time, and water quality in the harbor and bay. We set up and monitored multiple sites in both Buzzards Bay and West Falmouth Harbor, measuring depth, water velocity,salinity, pH, dissolved oxygen, chlorophyll-a, and nitrate concentration. In this report we present the processed time-series data at these locations and provide access to the data and metadata. The results will be used to understand circulation mechanisms and verify numerical models of hydrodynamics and biogeochemistry.

  13. Evaluation of Sediment Contamination in Pearl Harbor

    Science.gov (United States)

    1992-06-01

    ancient Hawaiians, was a large natural inland lagoon. Numerous walled fishponds located inside the harbor were used to cultivate various species of fishes... Ecotoxicology , Commission on Natural Resources, National Academy Press, Washington, D.C., 103 pp. National Research Council, 1989. Contaminated Marine

  14. Iodometric and Molecular Detection of ESBL Production Among Clinical Isolates of E. coli Fingerprinted by ERIC-PCR: The First Egyptian Report Declares the Emergence of E. coli O25b-ST131clone Harboring blaGES.

    Science.gov (United States)

    El-Badawy, Mohamed F; Tawakol, Wael M; Maghrabi, Ibrahim A; Mansy, Moselhy S; Shohayeb, Mohamed M; Ashour, Mohammed S

    2017-09-01

    The extensive use of β-lactam antibiotics has led to emergence and spread of extended-spectrum β-lactamases (ESBLs). This study was conducted to investigate the prevalence of 7 different ESBL genes (bla TEM , bla SHV , bla CTX-M , bla VEB , bla PER , bla GES , and bla OXA-10 ) and O25b-ST131 high-risk clone among 61 clinical isolates of Escherichia coli. Also, one broad-spectrum β-lactamase (bla OXA-1 ) was investigated. This study was also constructed to evaluate iodometric overlay method in detection of ESBL production. Phenotypic identification of E. coli isolates using API 20E revealed 18 distinct biotypes. DNA fingerprinting using enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) differentiated all isolates into 2 main phylogenetic groups with 60 distinct genetic profiles. Elevated values of minimal inhibitory concentration (MIC) 50 and MIC 90 for third- and fourth-generation cephalosporins were observed. Phenotypic tests revealed that 85.24% of isolates were ESBL producers. The incidence rates of bla TEM , bla SHV , bla CTX-M , bla GES , bla OXA-1 , and bla OXA-10 among E. coli ESBL producer phenotype were 69.23%, 25%, 96.15%, 3.85%, 11.54%, and 48%, respectively. On the other hand, bla VEB and bla PER were not detected. Sequencing of bla TEM and bla SHV revealed that bla TEM-214 and bla SHV-11 were the most prevalent variants. Group characterization of bla CTX-M revealed that bla CTX-M-1 was the most prevalent group of bla CTX-M family. It was found that 30.77% of E. coli ESBL producers belonged to O25b-ST131 clone harboring bla CTX-M-15 . This study concluded that iodometric overlay method was 100% sensitive in detection of ESBL production. To our knowledge, this is the first Egyptian study that declares the emergence of E. coli O25b-ST131 harboring bla GES .

  15. Increased level of extracellular ATP at tumor sites: in vivo imaging with plasma membrane luciferase.

    Directory of Open Access Journals (Sweden)

    Patrizia Pellegatti

    2008-07-01

    Full Text Available There is growing awareness that tumour cells build up a "self-advantageous" microenvironment that reduces effectiveness of anti-tumour immune response. While many different immunosuppressive mechanisms are likely to come into play, recent evidence suggests that extracellular adenosine acting at A2A receptors may have a major role in down-modulating the immune response as cancerous tissues contain elevated levels of adenosine and adenosine break-down products. While there is no doubt that all cells possess plasma membrane adenosine transporters that mediate adenosine uptake and may also allow its release, it is now clear that most of extracellularly-generated adenosine originates from the catabolism of extracellular ATP.Measurement of extracellular ATP is generally performed in cell supernatants by HPLC or soluble luciferin-luciferase assay, thus it generally turns out to be laborious and inaccurate. We have engineered a chimeric plasma membrane-targeted luciferase that allows in vivo real-time imaging of extracellular ATP. With this novel probe we have measured the ATP concentration within the tumour microenvironment of several experimentally-induced tumours.Our results show that ATP in the tumour interstitium is in the hundreds micromolar range, while it is basically undetectable in healthy tissues. Here we show that a chimeric plasma membrane-targeted luciferase allows in vivo detection of high extracellular ATP concentration at tumour sites. On the contrary, tumour-free tissues show undetectable extracellular ATP levels. Extracellular ATP may be crucial for the tumour not only as a stimulus for growth but also as a source of an immunosuppressive agent such as adenosine. Our approach offers a new tool for the investigation of the biochemical composition of tumour milieu and for development of novel therapies based on the modulation of extracellular purine-based signalling.

  16. Modeling of Tsunami Currents in Harbors

    Science.gov (United States)

    Lynett, P. J.

    2010-12-01

    Extreme events, such as large wind waves and tsunamis, are well recognized as a damaging hazard to port and harbor facilities. Wind wave events, particularly those with long period spectral components or infragravity wave generation, can excite resonance inside harbors leading to both large vertical motions and strong currents. Tsunamis can cause great damage as well. The geometric amplification of these very long waves can create large vertical motions in the interior of a harbor. Additionally, if the tsunami is composed of a train of long waves, which it often is, resonance can be easily excited. These long wave motions create strong currents near the node locations of resonant motions, and when interacting with harbor structures such as breakwaters, can create intense turbulent rotational structures, typical in the form of large eddies or gyres. These gyres have tremendous transport potential, and have been observed to break mooring lines, and even cause ships to be trapped inside the rotation, moving helplessly with the flow until collision, grounding, or dissipation of the eddy (e.g. Okal et al., 2006). This presentation will introduce the traditional theory used to predict wave impacts on harbors, discussing both how these models are practically useful and in what types of situations require a more accurate tool. State-of-the-art numerical models will be introduced, with a focus on recent developments in Boussinesq-type modeling. The Boussinesq equations model can account the dispersive, turbulent and rotational flow properties frequently observed in nature. Also they have the ability to coupling currents and waves and can predict nonlinear wave propagation over uneven bottom from deep (or intermediate) water area to shallow water area. However, during the derivation of a 2D-horizontal equation set, some 3D flow features, such those driven by as the dispersive stresses and the effects of the unresolved small scale 3D turbulence, are excluded. Consequently

  17. 32 CFR 765.6 - Regulations for Pearl Harbor, Hawaii.

    Science.gov (United States)

    2010-07-01

    ... 32 National Defense 5 2010-07-01 2010-07-01 false Regulations for Pearl Harbor, Hawaii. 765.6... RULES RULES APPLICABLE TO THE PUBLIC § 765.6 Regulations for Pearl Harbor, Hawaii. The Commander, U.S. Naval Base, Pearl Harbor, Hawaii, is responsible for prescribing and enforcing such rules and...

  18. Teaching about Pearl Harbor. Curriculum Enhancement Series #1.

    Science.gov (United States)

    Shields, Anna Marshall

    These materials consist of sample lesson plans for teaching about the Japanese attack on Pearl Harbor on December 7, 1941, in both U.S. and world history classes. The lesson plans challenge students to examine how current attitudes toward the Japanese may be rooted in World War II and Pearl Harbor. Selected bibliographies on Pearl Harbor, World…

  19. Useful Bicistronic Reporter System for Studying Poly(A Site-Defining cis Elements and Regulation of Alternative Polyadenylation

    Directory of Open Access Journals (Sweden)

    Zhongyuan Deng

    2018-01-01

    Full Text Available The link between polyadenylation (pA and various biological, behavioral, and pathological events of eukaryotes underlines the need to develop in vivo polyadenylation assay methods for characterization of the cis-acting elements, trans-acting factors and environmental stimuli that affect polyadenylation efficiency and/or relative usage of two alternative polyadenylation (APA sites. The current protein-based CAT or luciferase reporter systems can measure the polyadenylation efficiency of a single pA site or candidate cis element but not the choice of two APA sites. To address this issue, we developed a set of four new bicistronic reporter vectors that harbor either two luciferase or fluorescence protein open reading frames connected with one Internal Ribosome Entry Site (IRES. Transfection of single or dual insertion constructs of these vectors into mammalian cells demonstrated that they could be utilized not only to quantify the strength of a single candidate pA site or cis element, but also to accurately measure the relative usage of two APA sites at both the mRNA (qRT-PCR and protein levels. This represents the first reporter system that can study polyadenylation efficiency of a single pA site or element and regulation of two APA sites at both the mRNA and protein levels.

  20. Remembering Pearl Harbor at 75 Years.

    Science.gov (United States)

    Liehr, Patricia; Sopcheck, Janet; Milbrath, Gwyneth

    2016-12-01

    : On December 7, 1941, the Sunday-morning quiet of the U.S. naval base in Pearl Harbor, Hawaii, was shattered by dive-bombing Japanese fighter planes. The planes came in two waves-and when it was all over, more than 2,400 were killed and more than 1,100 were injured.Nurses were stationed at U.S. Naval Hospital Pearl Harbor, Tripler General Hospital (now Tripler Army Medical Center), Hickam Field Hospital, Schofield Barracks Station Hospital, and aboard the USS Solace, and witnessed the devastation. But they also did what nurses do in emergencies-they responded and provided care to those in need. Here are the stories of a few of those nurses.

  1. Solubilization and folding of a fully active recombinant Gaussia luciferase with native disulfide bonds by using a SEP-Tag.

    Science.gov (United States)

    Rathnayaka, Tharangani; Tawa, Minako; Nakamura, Takashi; Sohya, Shihori; Kuwajima, Kunihiro; Yohda, Masafumi; Kuroda, Yutaka

    2011-12-01

    Gaussia luciferase (GLuc) is the smallest known bioluminescent protein and is attracting much attention as a potential reporter protein. However, its 10 disulfide bond forming cysteines have hampered the efficient production of recombinant GLuc and thus limited its use in bio-imaging application. Here, we demonstrate that the addition of a short solubility enhancement peptide tag (SEP-Tag) to the C-terminus of GLuc (GLuc-C9D) significantly increased the fraction of soluble protein at a standard expression temperature. The expression time was much shorter, and the final yield of GLuc-C9D was significantly higher than with our previous pCold expression system. Reversed phase HPLC indicated that the GLuc-C9D variant folded with a single disulfide bond pattern after proper oxidization. Further, the thermal denaturation of GLuc-C9D was completely reversible, and its secondary structure content remained unchanged until 40°C as assessed by CD spectroscopy. The (1)H-NMR spectrum of GLuc indicated sharp well dispersed peaks typical for natively folded proteins. GLuc-C9D bioluminescence activity was strong and fully retained even after incubation at high temperatures. These results suggest that solubilization using SEP-Tags can be useful for producing large quantities of proteins containing multiple disulfide bonds. Copyright © 2011. Published by Elsevier B.V.

  2. In-vivo quantification of primary microRNA processing by Drosha with a luciferase based system

    International Nuclear Information System (INIS)

    Allegra, Danilo; Mertens, Daniel

    2011-01-01

    Research highlights: → Posttranscriptional regulation of miRNA processing is difficult to quantify. → Our in-vivo processing assay can quantify Drosha cleavage in live cells. → It is based on luciferase reporters fused with pri-miRNAs. → The assay validates the processing defect caused by a mutation in pri-16-1. → It is a sensitive method to quantify pri-miRNA cleavage by Drosha in live cells. -- Abstract: The RNAse III Drosha is responsible for the first step of microRNA maturation, the cleavage of primary miRNA to produce the precursor miRNA. Processing by Drosha is finely regulated and influences the amount of mature microRNA in a cell. We describe in the present work a method to quantify Drosha processing activity in-vivo, which is applicable to any microRNA. With respect to other methods for measuring Drosha activity, our system is faster and scalable, can be used with any cellular system and does not require cell sorting or use of radioactive isotopes. This system is useful to study regulation of Drosha activity in physiological and pathological conditions.

  3. Steady-state fluorescence and phosphorescence spectroscopic studies of bacterial luciferase tryptophan mutants.

    Science.gov (United States)

    Li, Z; Meighen, E A

    1994-09-01

    Bacterial luciferase, which catalyzes the bioluminescence reaction in luminous bacteria, consists of two nonidentical polypeptides, α and β. Eight mutants of luciferase with each of the tryptophans replaced by tyrosine were generated by site-directed mutagenesis and purified to homogeneity. The steady-state tryptophan fluorescence and low-temperature phosphorescence spectroscopic properties of these mutants were characterized. In some instances, mutation of only a single tryptophan residue resulted in large spectral changes. The tryptophan residues conserved in both the α and the β subunits exhibited distinct fluorescence emission properties, suggesting that these tryptophans have different local enviroments. The low-temperature phosphorescence data suggest that the tryptophans conserved in bot the α and the β subunits are not located at the subunit interface and/or involved in subunit interactions. The differences in the spectral properties of the mutants have provided useful information on the local environment of the individual tryptophan residues as well as on the quaternary structure of the protein.

  4. Highly specific expression of luciferase gene in lungs of naive nude mice directed by prostate-specific antigen promoter

    International Nuclear Information System (INIS)

    Li Hongwei; Li Jinzhong; Helm, Gregory A.; Pan Dongfeng

    2005-01-01

    PSA promoter has been demonstrated the utility for tissue-specific toxic gene therapy in prostate cancer models. Characterization of foreign gene overexpression in normal animals elicited by PSA promoter should help evaluate therapy safety. Here we constructed an adenovirus vector (AdPSA-Luc), containing firefly luciferase gene under the control of the 5837 bp long prostate-specific antigen promoter. A charge coupled device video camera was used to non-invasively image expression of firefly luciferase in nude mice on days 3, 7, 11 after injection of 2 x 10 9 PFU of AdPSA-Luc virus via tail vein. The result showed highly specific expression of the luciferase gene in lungs of mice from day 7. The finding indicates the potential limitations of the suicide gene therapy of prostate cancer based on selectivity of PSA promoter. By contrary, it has encouraging implications for further development of vectors via PSA promoter to enable gene therapy for pulmonary diseases

  5. Emergence of fatal avian influenza in New England harbor seals

    Science.gov (United States)

    Anthony, S.J.; St. Leger, J. A.; Pugliares, K.; Ip, Hon S.; Chan, J.M.; Carpenter, Z.W.; Navarrete-Macias, I.; Sanchez-Leon, M.; Saliki, J.T.; Pedersen, J.; Karesh, W.; Daszak, P.; Rabadan, R.; Rowles, T.; Lipkin, W.I.

    2012-01-01

    From September to December 2011, 162 New England harbor seals died in an outbreak of pneumonia. Sequence analysis of postmortem samples revealed the presence of an avian H3N8 influenza A virus, similar to a virus circulating in North American waterfowl since at least 2002 but with mutations that indicate recent adaption to mammalian hosts. These include a D701N mutation in the viral PB2 protein, previously reported in highly pathogenic H5N1 avian influenza viruses infecting people. Lectin staining and agglutination assays indicated the presence of the avian-preferred SAα-2,3 and mammalian SAα-2,6 receptors in seal respiratory tract, and the ability of the virus to agglutinate erythrocytes bearing either the SAα-2,3 or the SAα-2,6 receptor. The emergence of this A/harbor seal/Massachusetts/1/2011 virus may herald the appearance of an H3N8 influenza clade with potential for persistence and cross-species transmission.

  6. Phaeobacter inhibens from the Roseobacter clade has an environmental niche as a surface colonizer in harbors

    DEFF Research Database (Denmark)

    Gram, Lone; Rasmussen, Bastian Barker; Wemheuer, Bernd

    2015-01-01

    in the environment is to date unknown. Here, we report on the repeated isolation of Phaeobacter inhibens strains from a marine environment (harbors) not related to aquaculture. Based on phenotype and 16S rRNA gene sequence similarity, a total of 64 P. inhibens strains were identified from 35 samples (eukaryotic...... and used for amplification of 16S rRNA. P. inhibens was detected in five of these samples, all of which were biofilm samples, by pyrotag-sequencing at a prevalence of 0.02–0.68% of the prokaryotic community. The results indicated that P. inhibens had a niche in biofilms of fouled surfaces in harbor areas...

  7. Evaluation of the Luciferase Assay-Based In Vitro Elicitation Test for Serum IgE

    Directory of Open Access Journals (Sweden)

    Ryosuke Nakamura

    2012-01-01

    Results: The area under the ROC curves was highest in the EXiLE test (0.977, followed by CAP-FEIA (0.926 and degranulation (0.810. At an optimal cutoff range (1.648-1.876 calculated from the ROC curve of the EXiLE test, sensitivity and specificity were 0.944 and 0.917, respectively. A 95% positive predictive value was given at a cutoff level of 2.054 (fold increase in luciferase expression by logistic regression analysis. Conclusions: In contrast to in vivo tests, the EXiLE test appears to be a useful tool in diagnosing patients suspected of having IgE-dependent EW allergy without the risk of severe systemic reactions.

  8. Cultural Resources Survey of Mobile Harbor, Alabama.

    Science.gov (United States)

    1983-01-01

    improvement from the point of view of supply and communication with other European settlements, since it cut the lightering distance to the capital in half...order to cut the costs of building (Bathe 1978:08.00-02; Millar 1978:15-29). 32 6e The sharing of ship builders, the borrowing of vessel lines and the... Eslava Street Mobile. Burned to water’s edge during overhaul. Notes: Served as HINGHAM in Boston Harbor; served as ORIENT in Long Island Sound. Operated

  9. 75 FR 18887 - Division of Longshore and Harbor Workers' Compensation; Proposed Collection; Comment Request

    Science.gov (United States)

    2010-04-13

    ... can be provided in the desired format, reporting burden (time and financial resources) is minimized... comment on proposed and/or continuing collections of information in accordance with the paperwork... concerning the proposed collection: Securing Financial Obligations under the Longshore and Harbor Workers...

  10. The isolation and partial characterization of a highly pathogenic herpesvirus from the harbor seal (Phoca vitulina).

    NARCIS (Netherlands)

    A.D.M.E. Osterhaus (Albert); H. Yang (Hong); H.E.M. Spijkers (Ine); J. Groen (Jan); J.S. Teppema; G. van Steenis (Bert)

    1985-01-01

    textabstractThis report describes the first isolation and partial characterization of a herpesvirus from the harbor seal (Phoca vitulina). The virus was isolated during a disease outbreak in a group of young seals nursed in a seal orphanage in The Netherlands. Almost half of the seals died with

  11. 77 FR 50916 - Safety Zone; Boston Harbor's Rock Removal Project, Boston Inner Harbor, Boston, MA

    Science.gov (United States)

    2012-08-23

    ... operation on the navigable waters of Boston Inner Harbor, in the main ship channel near Castle Island. This... operations in support of the U.S. Army Corps of Engineers rock removal project. Entering into, transiting... before the start date of the project. The U.S. Army Corps of Engineers also discussed the rock removal...

  12. Using a split luciferase assay (SLA) to measure the kinetics of cell-cell fusion mediated by herpes simplex virus glycoproteins.

    Science.gov (United States)

    Saw, Wan Ting; Matsuda, Zene; Eisenberg, Roselyn J; Cohen, Gary H; Atanasiu, Doina

    2015-11-15

    Herpes simplex virus (HSV) entry and cell-cell fusion require the envelope proteins gD, gH/gL and gB. We propose that receptor-activated conformational changes to gD activate gH/gL, which then triggers gB (the fusogen) into an active form. To study this dynamic process, we have adapted a dual split protein assay originally developed to study the kinetics of human immunodeficiency virus (HIV) mediated fusion. This assay uses a chimera of split forms of renilla luciferase (RL) and green fluorescent protein (GFP). Effector cells are co-transfected with the glycoproteins and one of the split reporters. Receptor-bearing target cells are transfected with the second reporter. Co-culture results in fusion and restoration of RL, which can convert a membrane permeable substrate into a luminescent product, thereby enabling one to monitor initiation and extent of fusion in live cells in real time. Restoration of GFP can also be studied by fluorescence microscopy. Two sets of split reporters have been developed: the original one allows one to measure fusion kinetics over hours whereas the more recent version was designed to enhance the sensitivity of RL activity allowing one to monitor both initiation and rates of fusion in minutes. Here, we provide a detailed, step-by-step protocol for the optimization of the assay (which we call the SLA for split luciferase assay) using the HSV system. We also show several examples of the power of this assay to examine both the initiation and kinetics of cell-cell fusion by wild type forms of gD, gB, gH/gL of both serotypes of HSV as well as the effect of mutations and antibodies that alter the kinetics of fusion. The SLA can be applied to other viral systems that carry out membrane fusion. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. Human CRF{sub 2} {alpha} and {beta} splice variants: pharmacological characterization using radioligand binding and a luciferase gene expression assay

    Energy Technology Data Exchange (ETDEWEB)

    Ardati, A. [Rhone-Poulenc Rorer, Cardiovascular Biology, NW4, 500 Arcola Road, Collegeville, PA (United States); Goetschy, V.; Gottowick, J.; Henriot, S.; Deuschle, U.; Kilpatrick, G.J. [Central Nervous System, Pharma Division, F. Hoffmann-La Roche AG, CH-4070 Basel (Switzerland); Valdenaire, O. [Cardiovascular Research, Pharma Division, F. Hoffmann-La Roche AG, CH-4070 Basel (Switzerland)

    1999-03-14

    Corticotropin releasing factor (CRF) receptors belong to the super-family of G protein-coupled receptors. These receptors are classified into two subtypes (CRF{sub 1} and CRF{sub 2}). Both receptors are positively coupled to adenylyl cyclase but they have a distinct pharmacology and distribution in brain. Two isoforms belonging to the CRF{sub 2} subtype receptors, CRF{sub 2{alpha}} and CRF{sub 2{beta}}, have been identified in rat and man. The neuropeptides CRF and urocortin mediate their actions through this CRF G protein-coupled receptor family. In this report, we describe the pharmacological characterization of the recently identified hCRF{sub 2{beta}} receptor. We have used radioligand binding with [{sup 125}I]-tyr{sup 0}-sauvagine and a gene expression assay in which the firefly luciferase gene expression is under the control of cAMP responsive elements. Association kinetics of [{sup 125}I]-tyr{sup 0}-sauvagine binding to the hCRF{sub 2{beta}} receptor were monophasic while dissociation kinetics were biphasic, in agreement with the kinetics results obtained with the hCRF{sub 2{alpha}} receptor. Saturation binding analysis revealed two affinity states in HEK 293 cells with binding parameters in accord with those determined kinetically and with parameters obtained with the hCRF{sub 2{alpha}} receptor. A non-hydrolysable GTP analog, Gpp(NH)p, reduced the high affinity binding of [{sup 125}I]-tyr{sup 0}-sauvagine to both hCRF{sub 2} receptor isoforms in a similar manner. The rank order of potency of CRF agonist peptides in competition experiments was identical for both hCRF{sub 2}{alpha}-helical CRF{sub (9-41)}oCRF). Similarly, agonist potency was similar for the two isoforms when studied using the luciferase gene reporter system. The peptide antagonist {alpha}-helical CRF{sub (9-41)} exhibited a non-competitive antagonism of urocortin-stimulated luciferase expression with both hCRF{sub 2} receptor isoforms. Taken together, these results indicate that the

  14. Warm Water Bath Stimulates Phase-Shifts of the Peripheral Circadian Clocks in PER2::LUCIFERASE Mouse

    Science.gov (United States)

    Kuriki, Daisuke; Haraguchi, Atsushi; Shibata, Shigenobu

    2014-01-01

    Circadian clocks in the peripheral tissues of mice are known to be entrained by pulse stimuli such as restricted feeding, novel wheel running, and several other agents. However, there are no reports on high temperature pulse-mediated entrainment on the phase-shift of peripheral clocks in vivo. Here we show that temperature treatment of mice for two days at 41°C, instead of 37°C, for 1–2 h during the inactive period, using a temperature controlled water bath stimulated phase-advance of peripheral clocks in the kidney, liver, and submandibular gland of PER2::LUCIFERASE mice. On the other hand, treatment for 2 days at 35°C ambient room temperature for 2 h did not cause a phase-advance. Maintenance of mice at 41°C in a water bath, sustained the core body temperature at 40–41°C. However, the use of 37°C water bath or the 35°C ambient room temperature elevated the core body temperature to 38.5°C, suggesting that at least a core body temperature of 40–41°C is necessary to cause phase-advance under light-dark cycle conditions. The temperature pulse stimulation at 41°C, instead of 37°C water bath for 2 h led to the elevated expression of Per1 and Hsp70 in the peripheral tissue of mice. In summary, the present study demonstrates that transient high temperature pulse using water bath during daytime causes phase-advance in mouse peripheral clocks in vivo. The present results suggest that hot water bath may affect the phase of peripheral clocks. PMID:24933288

  15. Vibration-based structural health monitoring of harbor caisson structure

    Science.gov (United States)

    Lee, So-Young; Lee, So-Ra; Kim, Jeong-Tae

    2011-04-01

    This study presents vibration-based structural health monitoring method in foundation-structure interface of harbor caisson structure. In order to achieve the objective, the following approaches are implemented. Firstly, vibration-based response analysis method is selected and structural health monitoring (SHM) technique is designed for harbor caisson structure. Secondly, the performance of designed SHM technique for harbor structure is examined by FE analysis. Finally, the applicability of designed SHM technique for harbor structure is evaluated by dynamic tests on a lab-scaled caisson structure.

  16. Jam Formation of Traffic Flow in Harbor Tunnel

    International Nuclear Information System (INIS)

    He Hongdi; Lu Weizhen; Dong Liyun

    2011-01-01

    This paper reports a study concerning occurrence and growth of traffic jam in a harbor tunnel. The single-lane with three sections (downgrade, flat, and upgrade) is taken into account and they are characterized with different velocity limit. At the low density, the traffic current increases linearly with density and saturates at some values of immediately density. As the density increases, the traffic jam appears firstly before the upgrade section and then extends to the downgrade section. Additionally, the relationships of the velocity and headway against position in different densities are obtained from simulation. These results clearly clarify where and when the traffic jam appears. Finally, the critical densities are derived via the theoretical analysis before and after the discontinuous fronts and the theoretical results are consistent with the critical values of simulation results. (interdisciplinary physics and related areas of science and technology)

  17. Hazardous substances shipping at inland water harbors

    International Nuclear Information System (INIS)

    Benkovic, Z.

    2009-01-01

    Safety measures and regulations system covering the aspects of fire protection, professional and ecological safety are aimed to create a safe working environment, by detection and remedy of conditions that are potentially hazardous for the well-being of the employees or are leading to certain undesired events. Such unwanted incidents may result in different consequences: operating person's injury, environment pollution or material damage. This study attempts to illustrate the organization of work during hazardous matter loading and unloading at inland water harbors, based on legal provisions and decrees involving safety precautions, and in order to achieve constant enhancement of operating procedure, decreasing thereby the number of work-related injuries and various accidental situations. Fundamental precondition required to prevent possible accidents and to optimize general safety policy is to recognize and control any danger or potential hazard, as well as to be familiar with the legal provisions covering the inland waterway transport of harmful substances.(author)

  18. 33 CFR 80.1460 - Kahului Harbor, Maui, HI.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Kahului Harbor, Maui, HI. 80.1460 Section 80.1460 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY INTERNATIONAL NAVIGATION RULES COLREGS DEMARCATION LINES Pacific Islands § 80.1460 Kahului Harbor, Maui, HI. A line drawn...

  19. 33 CFR 80.1450 - Nawiliwili Harbor, Kauai, HI.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Nawiliwili Harbor, Kauai, HI. 80.1450 Section 80.1450 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY INTERNATIONAL NAVIGATION RULES COLREGS DEMARCATION LINES Pacific Islands § 80.1450 Nawiliwili Harbor, Kauai, HI...

  20. 33 CFR 110.238 - Apra Harbor, Guam.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Apra Harbor, Guam. 110.238 Section 110.238 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY ANCHORAGES ANCHORAGE REGULATIONS Anchorage Grounds § 110.238 Apra Harbor, Guam. (a) The anchorage grounds (Datum: WGS...

  1. 33 CFR 80.1470 - Kawaihae Harbor, Hawaii, HI.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Kawaihae Harbor, Hawaii, HI. 80.1470 Section 80.1470 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY INTERNATIONAL NAVIGATION RULES COLREGS DEMARCATION LINES Pacific Islands § 80.1470 Kawaihae Harbor, Hawaii, HI...

  2. 33 CFR 80.1480 - Hilo Harbor, Hawaii, HI.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Hilo Harbor, Hawaii, HI. 80.1480 Section 80.1480 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY INTERNATIONAL NAVIGATION RULES COLREGS DEMARCATION LINES Pacific Islands § 80.1480 Hilo Harbor, Hawaii, HI. A line drawn...

  3. 33 CFR 110.208 - Buffalo Harbor, N.Y.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Buffalo Harbor, N.Y. 110.208 Section 110.208 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY ANCHORAGES ANCHORAGE REGULATIONS Anchorage Grounds § 110.208 Buffalo Harbor, N.Y. (a) The anchorage grounds—(1...

  4. 32 CFR 705.31 - USS Arizona Memorial, Pearl Harbor.

    Science.gov (United States)

    2010-07-01

    ... 32 National Defense 5 2010-07-01 2010-07-01 false USS Arizona Memorial, Pearl Harbor. 705.31... NAVY REGULATIONS AND OFFICIAL RECORDS PUBLIC AFFAIRS REGULATIONS § 705.31 USS Arizona Memorial, Pearl Harbor. (a) Limited space and the desirability of keeping the Memorial simple and dignified require the...

  5. 76 FR 50489 - Agency Information Collection Activities: Harbor Maintenance Fee

    Science.gov (United States)

    2011-08-15

    ... Activities: Harbor Maintenance Fee AGENCY: U.S. Customs and Border Protection, Department of Homeland... Fee (CBP Forms 349 and 350). This is a proposed extension of an information collection that was... Fee. OMB Number: 1651-0055. Form Number: CBP Forms 349 and 350. Abstract: The Harbor Maintenance Fee...

  6. Autonomous bioluminescent expression of the bacterial luciferase gene cassette (lux in a mammalian cell line.

    Directory of Open Access Journals (Sweden)

    Dan M Close

    Full Text Available The bacterial luciferase (lux gene cassette consists of five genes (luxCDABE whose protein products synergistically generate bioluminescent light signals exclusive of supplementary substrate additions or exogenous manipulations. Historically expressible only in prokaryotes, the lux operon was re-synthesized through a process of multi-bicistronic, codon-optimization to demonstrate for the first time self-directed bioluminescence emission in a mammalian HEK293 cell line in vitro and in vivo.Autonomous in vitro light production was shown to be 12-fold greater than the observable background associated with untransfected control cells. The availability of reduced riboflavin phosphate (FMNH(2 was identified as the limiting bioluminescence substrate in the mammalian cell environment even after the addition of a constitutively expressed flavin reductase gene (frp from Vibrio harveyi. FMNH(2 supplementation led to a 151-fold increase in bioluminescence in cells expressing mammalian codon-optimized luxCDE and frp genes. When injected subcutaneously into nude mice, in vivo optical imaging permitted near instantaneous light detection that persisted independently for the 60 min length of the assay with negligible background.The speed, longevity, and self-sufficiency of lux expression in the mammalian cellular environment provides a viable and powerful alternative for real-time target visualization not currently offered by existing bioluminescent and fluorescent imaging technologies.

  7. Application of firefly luciferase assay for adenosine triphosphate (ATP) to antimicrobial drug sensitivity testing

    Science.gov (United States)

    Picciolo, G. L.; Tuttle, S. A.; Schrock, C. G.; Deming, J. W.; Barza, M. J.; Wienstein, L.; Chappelle, E. W.

    1977-01-01

    The development of a rapid method for determining microbial susceptibilities to antibiotics using the firefly luciferase assay for adenosine triphosphate (ATP) is documented. The reduction of bacterial ATP by an antimicrobial agent was determined to be a valid measure of drug effect in most cases. The effect of 12 antibiotics on 8 different bacterial species gave a 94 percent correlation with the standard Kirby-Buer-Agar disc diffusion method. A 93 percent correlation was obtained when the ATP assay method was applied directly to 50 urine specimens from patients with urinary tract infections. Urine samples were centrifuged first to that bacterial pellets could be suspended in broth. No primary isolation or subculturing was required. Mixed cultures in which one species was predominant gave accurate results for the most abundant organism. Since the method is based on an increase in bacterial ATP with time, the presence of leukocytes did not interfere with the interpretation of results. Both the incubation procedure and the ATP assays are compatible with automation.

  8. Probing Bioluminescence Resonance Energy Transfer in Quantum Rod-Luciferase Nanoconjugates.

    Science.gov (United States)

    Alam, Rabeka; Karam, Liliana M; Doane, Tennyson L; Coopersmith, Kaitlin; Fontaine, Danielle M; Branchini, Bruce R; Maye, Mathew M

    2016-02-23

    We describe the necessary design criteria to create highly efficient energy transfer conjugates containing luciferase enzymes derived from Photinus pyralis (Ppy) and semiconductor quantum rods (QRs) with rod-in-rod (r/r) microstructure. By fine-tuning the synthetic conditions, CdSe/CdS r/r-QRs were prepared with two different emission colors and three different aspect ratios (l/w) each. These were hybridized with blue, green, and red emitting Ppy, leading to a number of new BRET nanoconjugates. Measurements of the emission BRET ratio (BR) indicate that the resulting energy transfer is highly dependent on QR energy accepting properties, which include absorption, quantum yield, and optical anisotropy, as well as its morphological and topological properties, such as aspect ratio and defect concentration. The highest BR was found using r/r-QRs with lower l/w that were conjugated with red Ppy, which may be activating one of the anisotropic CdSe core energy levels. The role QR surface defects play on Ppy binding, and energy transfer was studied by growth of gold nanoparticles at the defects, which indicated that each QR set has different sites. The Ppy binding at those sites is suggested by the observed BRET red-shift as a function of Ppy-to-QR loading (L), where the lowest L results in highest efficiency and furthest shift.

  9. The location and nature of general anesthetic binding sites on the active conformation of firefly luciferase; a time resolved photolabeling study.

    Directory of Open Access Journals (Sweden)

    Sivananthaperumal Shanmugasundararaj

    Full Text Available Firefly luciferase is one of the few soluble proteins that is acted upon by a wide variety of general anesthetics and alcohols; they inhibit the ATP-driven production of light. We have used time-resolved photolabeling to locate the binding sites of alcohols during the initial light output, some 200 ms after adding ATP. The photolabel 3-azioctanol inhibited the initial light output with an IC50 of 200 µM, close to its general anesthetic potency. Photoincorporation of [(3H]3-azioctanol into luciferase was saturable but weak. It was enhanced 200 ms after adding ATP but was negligible minutes later. Sequencing of tryptic digests by HPLC-MSMS revealed a similar conformation-dependence for photoincorporation of 3-azioctanol into Glu-313, a residue that lines the bottom of a deep cleft (vestibule whose outer end binds luciferin. An aromatic diazirine analog of benzyl alcohol with broader side chain reactivity reported two sites. First, it photolabeled two residues in the vestibule, Ser-286 and Ile-288, both of which are implicated with Glu-313 in the conformation change accompanying activation. Second, it photolabeled two residues that contact luciferin, Ser-316 and Ser-349. Thus, time resolved photolabeling supports two mechanisms of action. First, an allosteric one, in which anesthetics bind in the vestibule displacing water molecules that are thought to be involved in light output. Second, a competitive one, in which anesthetics bind isosterically with luciferin. This work provides structural evidence that supports the competitive and allosteric actions previously characterized by kinetic studies.

  10. Assessment of a combination between hard structures and sand nourishment eastern of Damietta harbor using numerical modeling

    Directory of Open Access Journals (Sweden)

    A.M. Khalifa

    2017-12-01

    Full Text Available Damietta harbor was constructed in 1982 as an inland harbor with its 15 m depth navigation channel and two jetties acting like an obstacle to not allow sediment deposition in the harbor. On the other hand, they significantly affect the northern coast shoreline and hinder the sediment circulation in Damietta promontory. Satellite images show that new headlands are being implemented in the coastal shores of As-senaneyah. The proposed project consists of implementation of four headlands with length of 160 m, spacing of 400 m and using 150,000 m3 nourishment in those spacing between the hard structures only once during the construction time. Litpack 1D-model is used to predict shoreline responses to number of different five scenarios considered as combination between hard structures such as headlands and sand nourishment. A total number of 32 profiles were used to assess the shoreline changes along Gamasa, Damietta and Ras El-bar resort from 2010 to 2015. This study prevails a high erosion rate of the eastern and western shorelines of the proposed headlands. Nourishment of 200,000 m3/year is found to be a reasonable solution due to simplicity of being attained from Damietta harbor’s annual dredged materials which was reported to be average of 1 million m3/year. Keywords: Numerical modeling, Damietta harbor, Egyptian shoreline changes, Inland harbor

  11. 33 CFR 165.904 - Lake Michigan at Chicago Harbor & Burnham Park Harbor-Safety and Security Zone.

    Science.gov (United States)

    2010-07-01

    ... & Burnham Park Harbor-Safety and Security Zone. 165.904 Section 165.904 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PORTS AND WATERWAYS SAFETY REGULATED NAVIGATION... Guard District § 165.904 Lake Michigan at Chicago Harbor & Burnham Park Harbor—Safety and Security Zone...

  12. 77 FR 59551 - Safety Zone, Changes to Original Rule; Boston Harbor's Rock Removal Project, Boston Inner Harbor...

    Science.gov (United States)

    2012-09-28

    ... DEPARTMENT OF HOMELAND SECURITY Coast Guard 33 CFR Part 165 [Docket No. USCG-2012-0767] RIN 1625-AA00 Safety Zone, Changes to Original Rule; Boston Harbor's Rock Removal Project, Boston Inner Harbor... original provisions of that temporary final rule, but adds two additional safety zones necessary for the...

  13. Development of a dual luciferase activity and fluorescamine protein assay adapted to a 384 micro-well plate format: Reducing variability in human luciferase transactivation cell lines aimed at endocrine active substances

    Science.gov (United States)

    Brennan, Jennifer; Tillitt, Donald E.

    2018-01-01

    There is a need to adapt cell bioassays to 384-well and 1536-well formats instead of the traditional 96-well format as high-throughput screening (HTS) demands increase. However, the sensitivity and performance of the bioassay must be re-verified in these higher micro-well plates, and verification of cell health must also be HT (high-throughput). We have adapted two commonly used human breast luciferase transactivation cell bioassays, the recently re-named estrogen agonist/antagonist screening VM7Luc4E2 cell bioassay (previously designated BG1Luc4E2) and the androgen/glucocorticoid screening MDA-kb2 cell bioassay, to 384-well formats for HTS of endocrine-active substances (EASs). This cost-saving adaptation includes a fast, accurate, and easy measurement of protein amount in each well via the fluorescamine assay with which to normalize luciferase activity of cell lysates without requiring any transfer of the cell lysates. Here we demonstrate that by accounting for protein amount in the cell lysates, antagonistic agents can easily be distinguished from cytotoxic agents in the MDA-kb2 and VM7Luc4E2 cell bioassays. Additionally, we demonstrate via the fluorescamine assay improved interpretation of luciferase activity in wells along the edge of the plate (the so-called “edge effect”), thereby increasing usable wells to the entire plate, not just interior wells.

  14. Long-Term Management Strategy for Dredged Material Disposal for Naval Facilities at Pearl Harbor, Hawaii Phase III - Analysis of Alternatives and Development of an LTMS

    National Research Council Canada - National Science Library

    Palermo, Michael

    2000-01-01

    This report documents Phase III of a three-phase study to develop a Long-Term Management Study for disposal of dredged material unsuitable for ocean disposal from Pearl Harbor Naval Complex for the next 30 years...

  15. Wave Climate and Wave Response, 2025 Plan, Kahului Harbor, Maui, Hawaii

    National Research Council Canada - National Science Library

    Thompson, Edward

    2002-01-01

    ... (wind waves and swell) and long waves (harbor oscillations), was used to evaluate the technical feasibility of three alternative modifications to the harbor, including the Kahului Commercial Harbor 2025 Master Plan...

  16. The Evolution of the Bacterial Luciferase Gene Cassette (lux as a Real-Time Bioreporter

    Directory of Open Access Journals (Sweden)

    Gary Sayler

    2012-01-01

    Full Text Available The bacterial luciferase gene cassette (lux is unique among bioluminescent bioreporter systems due to its ability to synthesize and/or scavenge all of the substrate compounds required for its production of light. As a result, the lux system has the unique ability to autonomously produce a luminescent signal, either continuously or in response to the presence of a specific trigger, across a wide array of organismal hosts. While originally employed extensively as a bacterial bioreporter system for the detection of specific chemical signals in environmental samples, the use of lux as a bioreporter technology has continuously expanded over the last 30 years to include expression in eukaryotic cells such as Saccharomyces cerevisiae and even human cell lines as well. Under these conditions, the lux system has been developed for use as a biomedical detection tool for toxicity screening and visualization of tumors in small animal models. As the technologies for lux signal detection continue to improve, it is poised to become one of the first fully implantable detection systems for intra-organismal optical detection through direct marriage to an implantable photon-detecting digital chip. This review presents the basic biochemical background that allows the lux system to continuously autobioluminesce and highlights the important milestones in the use of lux-based bioreporters as they have evolved from chemical detection platforms in prokaryotic bacteria to rodent-based tumorigenesis study targets. In addition, the future of lux imaging using integrated circuit microluminometry to image directly within a living host in real-time will be introduced and its role in the development of dose/response therapeutic systems will be highlighted.

  17. Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation.

    Science.gov (United States)

    Li, Jian-Feng; Bush, Jenifer; Xiong, Yan; Li, Lei; McCormack, Matthew

    2011-01-01

    Protein-protein interactions (PPIs) constitute the regulatory network that coordinates diverse cellular functions. There are growing needs in plant research for creating protein interaction maps behind complex cellular processes and at a systems biology level. However, only a few approaches have been successfully used for large-scale surveys of PPIs in plants, each having advantages and disadvantages. Here we present split firefly luciferase complementation (SFLC) as a highly sensitive and noninvasive technique for in planta PPI investigation. In this assay, the separate halves of a firefly luciferase can come into close proximity and transiently restore its catalytic activity only when their fusion partners, namely the two proteins of interest, interact with each other. This assay was conferred with quantitativeness and high throughput potential when the Arabidopsis mesophyll protoplast system and a microplate luminometer were employed for protein expression and luciferase measurement, respectively. Using the SFLC assay, we could monitor the dynamics of rapamycin-induced and ascomycin-disrupted interaction between Arabidopsis FRB and human FKBP proteins in a near real-time manner. As a proof of concept for large-scale PPI survey, we further applied the SFLC assay to testing 132 binary PPIs among 8 auxin response factors (ARFs) and 12 Aux/IAA proteins from Arabidopsis. Our results demonstrated that the SFLC assay is ideal for in vivo quantitative PPI analysis in plant cells and is particularly powerful for large-scale binary PPI screens.

  18. Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation.

    Directory of Open Access Journals (Sweden)

    Jian-Feng Li

    Full Text Available Protein-protein interactions (PPIs constitute the regulatory network that coordinates diverse cellular functions. There are growing needs in plant research for creating protein interaction maps behind complex cellular processes and at a systems biology level. However, only a few approaches have been successfully used for large-scale surveys of PPIs in plants, each having advantages and disadvantages. Here we present split firefly luciferase complementation (SFLC as a highly sensitive and noninvasive technique for in planta PPI investigation. In this assay, the separate halves of a firefly luciferase can come into close proximity and transiently restore its catalytic activity only when their fusion partners, namely the two proteins of interest, interact with each other. This assay was conferred with quantitativeness and high throughput potential when the Arabidopsis mesophyll protoplast system and a microplate luminometer were employed for protein expression and luciferase measurement, respectively. Using the SFLC assay, we could monitor the dynamics of rapamycin-induced and ascomycin-disrupted interaction between Arabidopsis FRB and human FKBP proteins in a near real-time manner. As a proof of concept for large-scale PPI survey, we further applied the SFLC assay to testing 132 binary PPIs among 8 auxin response factors (ARFs and 12 Aux/IAA proteins from Arabidopsis. Our results demonstrated that the SFLC assay is ideal for in vivo quantitative PPI analysis in plant cells and is particularly powerful for large-scale binary PPI screens.

  19. A novel firefly luciferase biosensor enhances the detection of apoptosis induced by ESAT-6 family proteins of Mycobacterium tuberculosis

    International Nuclear Information System (INIS)

    Shi, Junwei; Zhang, Huan; Fang, Liurong; Xi, Yongqiang; Zhou, Yanrong; Luo, Rui; Wang, Dang; Xiao, Shaobo; Chen, Huanchun

    2014-01-01

    Highlights: • We developed a novel firefly luciferase based biosensor to detect apoptosis. • The novel biosensor 233-DnaE-DEVDG was reliable, sensitive and convenient. • 233-DnaE-DEVDG faithfully indicated ESAT-6 family proteins of Mycobacterium tuberculosis induced apoptosis. • EsxA, esxT and esxL in ESAT-6 family proteins induced apoptosis. • Activation of nuclear factor-κB (NF-κB) participated in esxT-induced apoptosis. - Abstract: The activation of caspase-3 is a key surrogate marker for detecting apoptosis. To quantitate caspase-3 activity, we constructed a biosensor comprising a recombinant firefly luciferase containing a caspase-3 cleavage site. When apoptosis was induced, caspase-3 cleavage of the biosensor activated firefly luciferase by a factor greater than 25. The assay conveniently detected apoptosis in real time, indicating that it will facilitate drug discovery. We screened ESAT-6 family proteins of Mycobacterium tuberculosis and found that esxA, esxT and esxL induced apoptosis. Further, activation of nuclear factor-κB (NF-κB) and the NF-κB-regulated genes encoding tumor necrosis factor-α (TNF-α) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) participated in esxT-induced apoptosis. We conclude that this assay is useful for high-throughput screening to identify and characterize proteins and drugs that regulate apoptosis

  20. A novel firefly luciferase biosensor enhances the detection of apoptosis induced by ESAT-6 family proteins of Mycobacterium tuberculosis

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Junwei; Zhang, Huan; Fang, Liurong; Xi, Yongqiang; Zhou, Yanrong; Luo, Rui; Wang, Dang, E-mail: wangdang511@126.com; Xiao, Shaobo; Chen, Huanchun

    2014-10-03

    Highlights: • We developed a novel firefly luciferase based biosensor to detect apoptosis. • The novel biosensor 233-DnaE-DEVDG was reliable, sensitive and convenient. • 233-DnaE-DEVDG faithfully indicated ESAT-6 family proteins of Mycobacterium tuberculosis induced apoptosis. • EsxA, esxT and esxL in ESAT-6 family proteins induced apoptosis. • Activation of nuclear factor-κB (NF-κB) participated in esxT-induced apoptosis. - Abstract: The activation of caspase-3 is a key surrogate marker for detecting apoptosis. To quantitate caspase-3 activity, we constructed a biosensor comprising a recombinant firefly luciferase containing a caspase-3 cleavage site. When apoptosis was induced, caspase-3 cleavage of the biosensor activated firefly luciferase by a factor greater than 25. The assay conveniently detected apoptosis in real time, indicating that it will facilitate drug discovery. We screened ESAT-6 family proteins of Mycobacterium tuberculosis and found that esxA, esxT and esxL induced apoptosis. Further, activation of nuclear factor-κB (NF-κB) and the NF-κB-regulated genes encoding tumor necrosis factor-α (TNF-α) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) participated in esxT-induced apoptosis. We conclude that this assay is useful for high-throughput screening to identify and characterize proteins and drugs that regulate apoptosis.

  1. Design and introduction of a disulfide bridge in firefly luciferase: increase of thermostability and decrease of pH sensitivity.

    Science.gov (United States)

    Imani, Mehdi; Hosseinkhani, Saman; Ahmadian, Shahin; Nazari, Mahboobeh

    2010-08-01

    The thermal sensitivity and pH-sensitive spectral properties of firefly luciferase have hampered its application in a variety of fields. It is proposed that the stability of a protein can be increased by introduction of disulfide bridge that decreases the configurational entropy of unfolding. A disulfide bridge is introduced into Photinus pyralis firefly luciferase to make two separate mutant enzymes with a single bridge. Even though the A103C/S121C mutant showed remarkable thermal stability, its specific activity decreased, whereas the A296C/A326C mutant showed tremendous thermal stability, relative pH insensitivity and 7.3-fold increase of specific activity. Moreover, the bioluminescence emission spectrum of A296C/A326C was resistant against higher temperatures (37 degrees C). Far-UV CD analysis showed slight secondary structure changes for both mutants. Thermal denaturation analysis showed that conformational stabilities of A103C/S121C and A296C/A326C are more than native firefly luciferase. It is proposed that since A296 and A326 are situated in the vicinity of the enzyme active site microenvironment in comparison with A103 and S121, the formation of a disulfide bridge in this region has more impact on enzyme kinetic characteristics.

  2. Bar Harbor, ME Tsunami Forecast Grids for MOST Model

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The Bar Harbor, Maine Forecast Model Grids provides bathymetric data strictly for tsunami inundation modeling with the Method of Splitting Tsunami (MOST) model. MOST...

  3. Hydrocarbon pollutants shape bacterial community assembly of harbor sediments

    KAUST Repository

    Barbato, Marta; Mapelli, Francesca; Magagnini, Mirko; Chouaia, Bessem; Armeni, Monica; Marasco, Ramona; Crotti, Elena; Daffonchio, Daniele; Borin, Sara

    2016-01-01

    Petroleum pollution results in co-contamination by different classes of molecules, entailing the occurrence of marine sediments difficult to remediate, as in the case of the Ancona harbor (Mediterranean Sea, Italy). Autochthonous bioaugmentation

  4. Extrachromosomal deoxyribonucleic acid in R factor-harboring Enterobacteriaceae

    DEFF Research Database (Denmark)

    Møller, JK; Bak, AL; Christiansen, C

    1976-01-01

    Extrachromosomal deoxyribonucleic acid (DNA) from 24 different R factor-harboring Enterobacteriaceae was isolated and characterized by analytical ultracentrifugation and electron microscopy. The R factors represented 15 different patterns of transferable drug resistance found in enterobacteria from...

  5. 75 FR 76613 - National Pearl Harbor Remembrance Day, 2010

    Science.gov (United States)

    2010-12-09

    .... The deadly attack on Pearl Harbor did not accomplish its mission of breaking the American spirit..., aiding the war effort by working in manufacturing plants, participating in rationing programs, or...

  6. Apra Harbor, Guam Tsunami Forecast Grids for MOST Model

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The Apra Harbor, Guam Forecast Model Grids provides bathymetric data strictly for tsunami inundation modeling with the Method of Splitting Tsunami (MOST) model. MOST...

  7. Pearl Harbor, Hawaii 1 arc-second DEM

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The 1/3-second Pearl Harbor Hawaii Elevation Grid provides bathymetric data in ASCII raster format of 1/3-second resolution in geographic coordinates. This grid is...

  8. Submarine Biofouling Control- Chlorination DATS Study at Pearl Harbor

    National Research Council Canada - National Science Library

    Wegand, John

    2001-01-01

    The intent of this document is to sumarize the chlorination studies performed at Naval Station, Pearl Harbor in support of biofouling control initiatives for the submarine community, as requested by NAVSEA 92T...

  9. Pearl Harbor, Hawaii Tsunami Forecast Grids for MOST Model

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The Pearl Harbor, Hawaii Forecast Model Grids provides bathymetric data strictly for tsunami inundation modeling with the Method of Splitting Tsunami (MOST) model....

  10. Establishment of a luciferase assay-based screening system: Fumitremorgin C selectively inhibits cellular proliferation of immortalized astrocytes expressing an active form of AKT

    International Nuclear Information System (INIS)

    Wang Lei; Sasai, Ken; Akagi, Tsuyoshi; Tanaka, Shinya

    2008-01-01

    The AKT pathway is frequently activated in glioblastoma, and as such, inhibitors of this pathway could prove very useful as anti-glioblastoma therapies. Here we established immortalized astrocytes expressing Renilla luciferase as well as those expressing both an active form of AKT and firefly luciferase. Since both luciferase activities represent the numbers of corresponding cell lines, novel inhibitors of the AKT pathway can be identified by treating co-cultures containing the two types of luciferase-expressing cells with individual compounds. Indeed, such a screening system succeeded in identifying fumitremorgin C as an efficient inhibitor of the AKT pathway, which was further confirmed by the ability of fumitremorgin C to selectively inhibit the growth of immortalized astrocytes expressing an active form of AKT. The present study proposes a broadly applicable approach for identifying therapeutic agents that target the pathways and/or molecules responsible for cancer development

  11. Harbor seal vibrissa morphology suppresses vortex-induced vibrations

    OpenAIRE

    Hanke, Wolf; Witte, Mathias; Miersch, Lars; Brede, Martin; Oeffner, Johannes; Michael, Mark; Hanke, Frederike; Leder, Alfred; Dehnhardt, Guido

    2010-01-01

    Harbor seals (Phoca vitulina) often live in dark and turbid waters, where their mystacial vibrissae, or whiskers, play an important role in orientation. Besides detecting and discriminating objects by direct touch, harbor seals use their whiskers to analyze water movements, for example those generated by prey fish or by conspecifics. Even the weak water movements left behind by objects that have passed by earlier can be sensed and followed accurately (hydrodynamic trail following). While scan...

  12. Tech Talk for Social Studies Teachers Lest We Forget: Remembering Pearl Harbor.

    Science.gov (United States)

    Green, Tim

    2001-01-01

    Presents an annotated bibliography that provides Web sites about Pearl Harbor (Hawaii). Includes Web sites that cover Pearl Harbor history, a live view of Pearl Harbor, stories from people who remember where they were during the attack, information on the naval station at Pearl Harbor, and a virtual tour of the USS Arizona. (CMK)

  13. 33 CFR 207.610 - St. Lawrence River, Cape Vincent Harbor, N.Y.; use, administration, and navigation of the harbor...

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false St. Lawrence River, Cape Vincent Harbor, N.Y.; use, administration, and navigation of the harbor and U.S. breakwater. 207.610 Section 207... NAVIGATION REGULATIONS § 207.610 St. Lawrence River, Cape Vincent Harbor, N.Y.; use, administration, and...

  14. Characteristics and management of Enterobacteriaceae harboring IMP-4 or IMP-8 carbapenemase in a tertiary hospital.

    Science.gov (United States)

    Pang, Feng; Jia, Xiu-Qin; Song, Zhen-Zhu; Li, Yan-Hua; Wang, Bin; Zhao, Qi-Gang; Wang, Chuan-Xin; Zhang, Yi; Wang, Le-Xin

    2016-03-01

    The emergence of Enterobacteriaceae harboring IMP-4 or IMP-8 carbapenemases is rare. We report an occurrence of Enterobacteriaceae harboring IMP-4 or IMP-8 carbapenemases in a Chinese tertiary care hospital from November 2010 to December 2012. The clinical characteristics of 30 patients were described. The genetic relationship of isolates was determined by pulsed-field gel electrophoresis (PFGE). Carbapenemases were detected by modified Hodge test (MHT) and polymerase chain reactions (PCRs). Amplicons were sequenced and blasted to determine the genotype. Most infected patients were from intensive care unit and had complex and serious underlying illnesses requiring mechanical ventilation. PFGE revealed that Klebsiella pneumoniae showed two major PFGE types. Two Klebsiella oxytoca had an indistinguishable PFGE pattern, while four Enterobacter cloacae were different strains. The sequencing studies showed Enterobacteriaceae harboring IMP-4 or IMP-8 carbapenemase in the 23 infected patients. The majority of patients had infections with the carbapenemase-producing Enterobacteriaceae (CPE) strain, most were successfully treated with a range of antibiotics and discharged. It is important to maintain a high index of suspicion to screen for carbapenemase-producing Enterobacteriaceae strains. Rapid identification of these strains and implementation of stringent procedures are the key to prevent major outbreaks in a hospital setting.

  15. Organochlorines in harbor seal (Phoca vitulina) tissues from the northern Gulf of Alaska

    International Nuclear Information System (INIS)

    Wang Dongli; Atkinson, Shannon; Hoover-Miller, Anne; Lee, Sung-Eun; Li, Qing X.

    2007-01-01

    Polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolites, heptachlor and hexachlorocyclohexanes (HCHs) were analyzed in the harbor seal (Phoca vitulina) tissues collected from the Gulf of Alaska during 2000-2001. ΣPCBs (16-728 ng/g lw) and ΣDDTs (14-368 ng/g lw) were the predominant pollutants followed by ΣHCHs (0.56-93 ng/g lw) and heptachlor (≤0.068-6.0 ng/g lw). Concentrations of the above organochlorines (OCs) in the liver, kidney and blubber tissues correlated with ages, sex, body weight and blubber thickness of the harbor seals. The OC concentrations were similar between the samples collected from two different regions - Prince William Sound, and Kodiak Island and Southern Alaska Peninsula. Mean levels of OCs in nursing female adults were much lower than those in male adults, which indicate that lactation transfer OCs from mother seals to newborns. - Σ28 PCB congeners, p,p'-DDE, p,p'-DDD and p,p'-DDT, α-, β-, γ-, and δ-HCH and heptachlor in harbor seal tissues from the northern Gulf of Alaska are reported

  16. Gene silencing in primary and metastatic tumors by small interfering RNA delivery in mice: quantitative analysis using melanoma cells expressing firefly and sea pansy luciferases.

    Science.gov (United States)

    Takahashi, Yuki; Nishikawa, Makiya; Kobayashi, Naoki; Takakura, Yoshinobu

    2005-07-20

    Silencing of oncogenes or other genes contributing to tumor malignancy or progression by RNA interference (RNAi) offers a promising approach to treating tumor patients. To achieve RNAi-based tumor therapy, a small interfering RNA (siRNA) or siRNA-expressing vector needs to be delivered to tumor cells, but little information about its in vivo delivery has been reported. In this study, we examined whether the expression of the target gene in tumor cells can be suppressed by the delivery of RNAi effectors to primary and metastatic tumor cells. To quantitatively evaluate the RNAi effects in tumor cells, mouse melanoma B16-BL6 cells were stably transfected with both firefly (a model target gene) and sea pansy (an internal standard gene) luciferase genes to obtain B16-BL6/dual Luc cells. The target gene expression in subcutaneous primary tumors of B16-BL6/dual Luc cells was significantly suppressed by direct injection of the RNAi effectors followed by electroporation. The expression in metastatic hepatic tumors was also significantly reduced by an intravenous injection of either RNAi effector by the hydrodynamics-based procedure. These results indicate that the both RNAi effectors have a potential to silence target gene in tumor cells in vivo when successfully delivered to tumor cells.

  17. Littoral cells and harbor dredging along the California coast

    Science.gov (United States)

    Griggs, Gary B.

    1987-02-01

    Beach compartments or littoral cells form the framework for our understanding of the sources, transport, and sinks of sand in the nearshore zone. In general, along the California coast, beach sand is derived from rivers or clifferosion, moves alongshore under the influence of the prevailing waver conditions, and ultimately is lost either to a submarine canyon or a dune field. Marinas or harbors built either between or at the upcoast ends of beach compartments have been relatively maintenance-free, because of a lack of significant littoral drift at these locations. On the other hand, those harbors built in the middle reaches or at the downcoast ends of littoral cells have had expensive annual dredging problems, because of the interruption of large volumes of littoral drift. Although engineers have labored for years on various breakwater, jetty, or entrance channel configurations, the actual design utilized is of secondary importance. The critical factors are harbor location within a littoral cell and annual litoral drift volume.

  18. Respiratory properties of blood in the harbor porpoise, Phocoena phocoena

    DEFF Research Database (Denmark)

    Soegaard, Lisette B; Hansen, Marie N; van Elk, Cornelis

    2012-01-01

    Harbor porpoises are active divers that exchange O(2) and CO(2) with the environment during a fast single breath upon surfacing. We investigated blood O(2)-transporting properties, buffer characteristics, Cl(-) transport via the erythrocyte anion exchanger (AE1), circulating nitric oxide metaboli......Harbor porpoises are active divers that exchange O(2) and CO(2) with the environment during a fast single breath upon surfacing. We investigated blood O(2)-transporting properties, buffer characteristics, Cl(-) transport via the erythrocyte anion exchanger (AE1), circulating nitric oxide....... The true plasma non-bicarbonate buffer value was moderately higher than in terrestrial mammals and increased upon deoxygenation. Plasma bicarbonate was also relatively high, contributing to increase the overall buffer capacity. The apparent Cl(-) permeability of harbor porpoise erythrocytes was similar......, plasma nitrate and hemoglobin-mediated nitrite reduction were elevated compared with mammalian standards, suggesting that increased nitric oxide bioavailability and nitrite-derived nitric oxide could play important roles in diving physiology....

  19. Engineering the metal sensitive sites in Macrolampis sp2 firefly luciferase and use as a novel bioluminescent ratiometric biosensor for heavy metals.

    Science.gov (United States)

    Gabriel, Gabriele V M; Viviani, Vadim R

    2016-12-01

    Most luminescent biosensors for heavy metals are fluorescent and rely on intensity measurements, whereas a few are ratiometric and rely on spectral changes. Bioluminescent biosensors for heavy metals are less common. Firefly luciferases have been coupled to responsive promoters for mercury and arsenium, and used as light on biosensors. Firefly luciferase bioluminescence spectrum is naturally sensitive to heavy metal cations such as zinc and mercury and to pH. Although pH sensitivity of firefly luciferases was shown to be useful for ratiometric estimation of intracellular pH, its potential use for ratiometric estimation of heavy metals was never considered. Using the yellow-emitting Macrolampis sp2 firefly luciferase and site-directed mutagenesis, we show that the residues H310 and E354 constitute two critical sites for metal sensitivity that can be engineered to increase sensitivity to zinc, nickel, and mercury. A linear relationship between cation concentration and the ratio of bioluminescence intensities at 550 and 610 nm allowed, for the first time, the ratiometric estimation of heavy metals concentrations down to 0.10 mM, demonstrating the potential applicability of firefly luciferases as enzymatic and intracellular ratiometric metal biosensors.

  20. 33 CFR 110.87 - Henderson Harbor, N.Y.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Henderson Harbor, N.Y. 110.87... ANCHORAGE REGULATIONS Special Anchorage Areas § 110.87 Henderson Harbor, N.Y. (a) Area A. The area in the... latitude 43°51′08.8″ N, longitude 76°12′08.9″ W, thence to latitude 43°51′09.0″ N, longitude 76°12′19.0″ W...

  1. Extrachromosomal deoxyribonucleic acid in R factor-harboring Enterobacteriaceae

    DEFF Research Database (Denmark)

    Møller, JK; Bak, AL; Christiansen, C

    1976-01-01

    Extrachromosomal deoxyribonucleic acid (DNA) from 24 different R factor-harboring Enterobacteriaceae was isolated and characterized by analytical ultracentrifugation and electron microscopy. The R factors represented 15 different patterns of transferable drug resistance found in enterobacteria from...... from 1.700 to 1.720 g/cm3. The majority of the bacteria contained extrachromosomal DNAs of various densities. Three-fourths of the R factors were classified as fi+. The investigation illustrates the extensive variability in the physical characteristics of plasmid DNA from R factor-harboring strains....

  2. Characteristics and management of Enterobacteriaceae harboring ...

    African Journals Online (AJOL)

    Patients who are hospitalized for prolonged periods and those with severe ... Hospital ICU, one case in PICU and one case in surgical. ICU. Other five cases ..... specificity to detect KPC but still uncertain to Ambler B ... be attributed to the high level AmpC or ESBLs enzymes ... This problem has been reported pre- viously33.

  3. Tales of some ancient harbors in the Aegean back-arc region: Earthquakes, coastal changes, historical impacts

    Science.gov (United States)

    Stiros, Stathis; Saltogianni, Vasso

    2017-04-01

    Tectonically active terrains are characterized by seismic transient ground motions (shaking) and by permanent ground motions in the vicinity of activated faults, with both effects occasionally leaving their signature on human constructions and the landscape. Especially in coastal areas marked by small tidal ranges and normal water salinity, as is the case with most parts of the Eastern Mediterranean, even small-amplitude tectonic motions can be derived from observations on coastal constructions, mainly harbors, but also on spring chambers in nearly arid environments, sewers, etc. Such observations, if coupled with well-dated observations of destructions and repairs and of changes in the occupation style of ancient sites can permit precious information conveyed from archaeology to tectonics/seismology and vice versa. A transect with harbor remains from Rhodes to the Gulf of Corinth and then till the Ionian Islands provides some excellent examples. The military harbor of Rhodes, in an area of long term uplift, a coded report of which seems to be provided by ancient poet Pindar, was subject to seismic subsidence and destruction, but with major international support, it was repaired, till renewed uplift brought it several meters above the water. In the Corinth area, the Kenchreai harbor was abruptly submerged during a major repair of a temple, as revealed by precious stained glass panels, ready to use but abandoned in shallow water beneath ruins, while radiometric dating of the uplift in the western Lechaion harbor, constrains its excavation in swampy environment not in Roman times, but to the period of flourishing of Corinth in circa 600BC and the colonization of Italy. Farther west, the sea-level mark of the harbor of Aigeira, at Mavra Litharia (Derveni/Akrata) indicates 4m uplift since the Roman period, at least partly seismic, correlating with an exposed reef and the abandonment of the repairs of the theatre of Aigeira. Seismic land uplift explains the demise of

  4. Harbor seal vibrissa morphology suppresses vortex-induced vibrations.

    Science.gov (United States)

    Hanke, Wolf; Witte, Matthias; Miersch, Lars; Brede, Martin; Oeffner, Johannes; Michael, Mark; Hanke, Frederike; Leder, Alfred; Dehnhardt, Guido

    2010-08-01

    Harbor seals (Phoca vitulina) often live in dark and turbid waters, where their mystacial vibrissae, or whiskers, play an important role in orientation. Besides detecting and discriminating objects by direct touch, harbor seals use their whiskers to analyze water movements, for example those generated by prey fish or by conspecifics. Even the weak water movements left behind by objects that have passed by earlier can be sensed and followed accurately (hydrodynamic trail following). While scanning the water for these hydrodynamic signals at a swimming speed in the order of meters per second, the seal keeps its long and flexible whiskers in an abducted position, largely perpendicular to the swimming direction. Remarkably, the whiskers of harbor seals possess a specialized undulated surface structure, the function of which was, up to now, unknown. Here, we show that this structure effectively changes the vortex street behind the whiskers and reduces the vibrations that would otherwise be induced by the shedding of vortices from the whiskers (vortex-induced vibrations). Using force measurements, flow measurements and numerical simulations, we find that the dynamic forces on harbor seal whiskers are, by at least an order of magnitude, lower than those on sea lion (Zalophus californianus) whiskers, which do not share the undulated structure. The results are discussed in the light of pinniped sensory biology and potential biomimetic applications.

  5. Integrated approach to assess ecosystem health in harbor areas.

    Science.gov (United States)

    Bebianno, M J; Pereira, C G; Rey, F; Cravo, A; Duarte, D; D'Errico, G; Regoli, F

    2015-05-01

    Harbors are critical environments with strategic economic importance but with potential environmental impact: health assessment criteria are a key issue. An ecosystem health status approach was carried out in Portimão harbor as a case-study. Priority and specific chemical levels in sediments along with their bioavailability in mussels, bioassays and a wide array of biomarkers were integrated in a biomarker index (IBR index) and the overall data in a weight of evidence (WOE) model. Metals, PAHs, PCBs and HCB were not particularly high compared with sediment guidelines and standards for dredging. Bioavailability was evident for Cd, Cu and Zn. Biomarkers proved more sensitive namely changes of antioxidant responses, metallothioneins and vittellogenin-like proteins. IBR index indicated that site 4 was the most impacted area. Assessment of the health status by WOE approach highlighted the importance of integrating sediment chemistry, bioaccumulation, biomarkers and bioassays and revealed that despite some disturbance in the harbor area, there was also an impact of urban effluents from upstream. Environmental quality assessment in harbors. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. 78 FR 63381 - Safety Zones; Hawaiian Island Commercial Harbors, HI

    Science.gov (United States)

    2013-10-24

    ...; Kaumalapau, Lanai; Kahului, Maui and Kawaihae and Hilo on the Island of Hawaii). The purpose of these safety... the piers faces. (9) All waters of Hilo Harbor, Hawaii immediately adjacent to commercial piers 1 and.... SUMMARY: The Coast Guard is establishing nine (9) permanent safety zones encompassing Hawaii's commercial...

  7. 33 CFR 117.181 - Oakland Inner Harbor Tidal Canal.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Oakland Inner Harbor Tidal Canal. 117.181 Section 117.181 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY... Tidal Canal. The draws of the Alameda County highway drawbridges at Park Street, mile 5.2; Fruitvale...

  8. 78 FR 21597 - Marine Mammals: Alaska Harbor Seal Habitats

    Science.gov (United States)

    2013-04-11

    ... seek input as to whether management measures are needed, and if so, what types of measures should be... proposed rulemaking (ANPR) on potential management measures to protect glacially-associated harbor seal... need for regulations; (2) the geographic scope and time horizon of regulations; (3) management options...

  9. Quantitative Measures of Anthropogenic Noise on Harbor Porpoises

    DEFF Research Database (Denmark)

    Wisniewska, Danuta Maria; Teilmann, Jonas; Hermannsen, Line

    2016-01-01

    -animal recordings as proxies for actual exposure. Here, we quantify sound exposure levels recorded with a DTAG-3 tag on a captive harbor porpoise exposed to vessel noise in a controlled acoustic environment. Results show that fl ow noise is limiting onboard noise recordings, whereas no evidence of body shading has...

  10. Rhizosphere of rice plants harbor bacteria with multiple plant growth ...

    African Journals Online (AJOL)

    Rhizosphere of rice plants harbor bacteria with multiple plant growth promoting features. ... 45 (39.46%) isolates were capable of producing siderophore, the range of production being 4.50 to 223.26 μg mg-1 protein. Analysis of molecular diversity was made by amplified ribosomal DNA restriction analysis (ARDRA) and ...

  11. U.S. Department of Defense - Pearl Harbor Special

    Science.gov (United States)

    Hawaii saw that seminal moment in history, and those that were there vividly remember that Sunday morning this noise," he said in an oral history on the Pearl Harbor Survivors' Association Web site. " seeing was an attack. "It didn't mean anything to us until a large group of planes came near the

  12. Constant pH Accelerated Molecular Dynamics Investigation of the pH Regulation Mechanism of Dinoflagellate Luciferase.

    Science.gov (United States)

    Donnan, Patrick H; Ngo, Phong D; Mansoorabadi, Steven O

    2018-01-23

    The bioluminescence reaction in dinoflagellates involves the oxidation of an open-chain tetrapyrrole by the enzyme dinoflagellate luciferase (LCF). The activity of LCF is tightly regulated by pH, where the enzyme is essentially inactive at pH ∼8 and optimally active at pH ∼6. Little is known about the mechanism of LCF or the structure of the active form of the enzyme, although it has been proposed that several intramolecularly conserved histidine residues in the N-terminal region are important for the pH regulation mechanism. Here, constant pH accelerated molecular dynamics was employed to gain insight into the conformational activation of LCF induced by acidification.

  13. A rapid method for the determination of microbial susceptibility using the firefly luciferase assay for adenosine triphosphate (ATP)

    Science.gov (United States)

    Vellend, H.; Tuttle, S. A.; Barza, M.; Weinstein, L.; Picciolo, G. L.; Chappelle, E. W.

    1975-01-01

    Luciferase assay for adenosine triphosphate (ATP) was optimized for pure bacteria in broth in order to evaluate if changes in bacterial ATP content could be used as a rapid measure of antibiotic effect on microorganisms. Broth cultures of log phase bacteria were incubated at 310 K (37 C) for 2.5 hours at antimicrobial concentrations which resulted in the best discrimination between sensitive and resistant strains. Eighty-seven strains of 11 bacterial species were studied for their susceptibility to 12 commonly used antimicrobial agents: ampicillin, Penicillin G, nafcillin, carbenicillin, cephalothin, tetracycline, erythromycin, clindamycin, gentamicin, nitrofurantoin, colistin, and chloramplenicol. The major advantage of the ATP system over existing methods of rapid microbial susceptibility testing is that the assay can be made specific for bacterial ATP.

  14. DNA repair in human fibroblasts, as reflected by host-cell reactivation of a transfected UV-irradiated luciferase gene, is not related to donor age

    International Nuclear Information System (INIS)

    Merkle, Thomas J.; O'Brien, Katherine; Brooks, Philip J.; Tarone, Robert E.; Robbins, Jay H.

    2004-01-01

    The effect of donor age on the ability of mammalian cells to repair ultraviolet (UV)-induced DNA damage has been studied using several approaches, most recently via assays that measure the host-cell reactivation (HCR) of UV-irradiated reporter gene-containing plasmid vectors following their transfection into cells. Plasmid HCR assays indirectly quantify a cell line's ability to perform nucleotide excision repair (NER) by measuring the enzyme activity of the repaired reporter gene, e.g., chloramphenical acetyltransferase (cat) or luciferase (luc), and are useful in studies investigating whether increasing age may be a risk factor for the deficient repair of potentially cancer-causing, sunlight-induced, DNA lesions in skin cells. In our study, we quantified the DNA repair ability of cultured, nontransformed, human skin fibroblast lines through their HCR of a transfected UV-C-irradiated plasmid containing luc. HCR was measured at various times after transfection in five lines from normal donors of ages 21-96 years, and from one donor who had xeroderma pigmentosum (XP). The normal lines displayed increasing HCR at successive post-transfection time points and showed no significant correlation between HCR and donor age. The XP-A line, known to be markedly deficient in NER of UV-induced DNA damage, showed minimal evidence of HCR compared to the normal lines. To further assess potential variation in HCR with donor age, fibroblast lines from five old donors, ages 84-94 years, were compared with lines from five young donors, ages 17-26 years. While significant differences in HCR were found between some lines, no significant difference was found between the young and old age groups (P=0.44). Our study provides no indication that the higher incidence of skin cancer observed with increasing age is due to an age-related decrease in the ability to repair UV-induced DNA damage

  15. DNA repair in human fibroblasts, as reflected by host-cell reactivation of a transfected UV-irradiated luciferase gene, is not related to donor age

    Energy Technology Data Exchange (ETDEWEB)

    Merkle, Thomas J.; O' Brien, Katherine; Brooks, Philip J.; Tarone, Robert E.; Robbins, Jay H

    2004-10-04

    The effect of donor age on the ability of mammalian cells to repair ultraviolet (UV)-induced DNA damage has been studied using several approaches, most recently via assays that measure the host-cell reactivation (HCR) of UV-irradiated reporter gene-containing plasmid vectors following their transfection into cells. Plasmid HCR assays indirectly quantify a cell line's ability to perform nucleotide excision repair (NER) by measuring the enzyme activity of the repaired reporter gene, e.g., chloramphenical acetyltransferase (cat) or luciferase (luc), and are useful in studies investigating whether increasing age may be a risk factor for the deficient repair of potentially cancer-causing, sunlight-induced, DNA lesions in skin cells. In our study, we quantified the DNA repair ability of cultured, nontransformed, human skin fibroblast lines through their HCR of a transfected UV-C-irradiated plasmid containing luc. HCR was measured at various times after transfection in five lines from normal donors of ages 21-96 years, and from one donor who had xeroderma pigmentosum (XP). The normal lines displayed increasing HCR at successive post-transfection time points and showed no significant correlation between HCR and donor age. The XP-A line, known to be markedly deficient in NER of UV-induced DNA damage, showed minimal evidence of HCR compared to the normal lines. To further assess potential variation in HCR with donor age, fibroblast lines from five old donors, ages 84-94 years, were compared with lines from five young donors, ages 17-26 years. While significant differences in HCR were found between some lines, no significant difference was found between the young and old age groups (P=0.44). Our study provides no indication that the higher incidence of skin cancer observed with increasing age is due to an age-related decrease in the ability to repair UV-induced DNA damage.

  16. A luciferase-based assay for rapid assessment of drug activity against Mycobacterium tuberculosis including monitoring of macrophage viability.

    Science.gov (United States)

    Larsson, Marie C; Lerm, Maria; Ängeby, Kristian; Nordvall, Michaela; Juréen, Pontus; Schön, Thomas

    2014-11-01

    The intracellular (IC) effect of drugs against Mycobacterium tuberculosis (Mtb) is not well established but increasingly important to consider when combining current and future multidrug regimens into the best possible treatment strategies. For this purpose, we developed an IC model based on a genetically modified Mtb H37Rv strain, expressing the Vibrio harvei luciferase (H37Rv-lux) infecting the human macrophage like cell line THP-1. Cells were infected at a low multiplicity of infection (1:1) and subsequently exposed to isoniazid (INH), ethambutol (EMB), amikacin (AMI) or levofloxacin (LEV) for 5days in a 96-well format. Cell viability was evaluated by Calcein AM and was maintained throughout the experiment. The number of viable H37Rv-lux was determined by luminescence and verified by a colony forming unit analysis. The results were compared to the effects of the same drugs in broth cultures. AMI, EMB and LEV were significantly less effective intracellularly (MIC90: >4mg/L, 8mg/L and 2mg/L, respectively) compared to extracellularly (MIC90: 0.5mg/L for AMI and EMB; 0.25mg/L for LEV). The reverse was the case for INH (IC: 0.064mg/L vs EC: 0.25mg/L). In conclusion, this luciferase based method, in which monitoring of cell viability is included, has the potential to become a useful tool while evaluating the intracellular effects of anti-mycobacterial drugs. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Underwater noise from three types of offshore wind turbines: estimation of impact zones for harbor porpoises and harbor seals.

    Science.gov (United States)

    Tougaard, Jakob; Henriksen, Oluf Damsgaard; Miller, Lee A

    2009-06-01

    Underwater noise was recorded from three different types of wind turbines in Denmark and Sweden (Middelgrunden, Vindeby, and Bockstigen-Valar) during normal operation. Wind turbine noise was only measurable above ambient noise at frequencies below 500 Hz. Total sound pressure level was in the range 109-127 dB re 1 microPa rms, measured at distances between 14 and 20 m from the foundations. The 1/3-octave noise levels were compared with audiograms of harbor seals and harbor porpoises. Maximum 1/3-octave levels were in the range 106-126 dB re 1 microPa rms. Maximum range of audibility was estimated under two extreme assumptions on transmission loss (3 and 9 dB per doubling of distance, respectively). Audibility was low for harbor porpoises extending 20-70 m from the foundation, whereas audibility for harbor seals ranged from less than 100 m to several kilometers. Behavioral reactions of porpoises to the noise appear unlikely except if they are very close to the foundations. However, behavioral reactions from seals cannot be excluded up to distances of a few hundred meters. It is unlikely that the noise reaches dangerous levels at any distance from the turbines and the noise is considered incapable of masking acoustic communication by seals and porpoises.

  18. Aerial Survey Counts of Harbor Seals in Coastal Alaska (2003-2011)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This dataset supports efforts to estimate the abundance and trends in population size of Alaska harbor seals. Annual surveys of harbor seal populations are...

  19. Aerial Survey Counts of Harbor Seals in Coastal Alaska (1998-2002)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This dataset supports efforts to estimate the abundance and trends in population size of Alaska harbor seals. Annual surveys of harbor seal populations are...

  20. A Comprehensive Copper Compliance Strategy: Implementing Regulatory Guidance at Pearl Harbor Naval Shipyard & Intermediate Maintenance Facility

    National Research Council Canada - National Science Library

    Earley, P. J; Rosen, G; Rivera-Duarte, I; Gauthier, R. D; Arias-Thode, Y; Thompson, J; Swope, B

    2007-01-01

    Studies were performed to develop a new National Pollution Discharge Elimination Systems Permit for the discharge of effluents from the Pearl Harbor Naval Shipyard and Intermediate Maintenance Facility into Pearl Harbor...

  1. Telemetry data from satellite tags deployed on harbor seals in Cook Inlet, Alaska

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Between 2004 and 2006 we conducted four harbor seal tagging trips in Cook Inlet during the months of October and May. In total, we captured and released 93 harbor...

  2. A smart ROV solution for ship hull and harbor inspection

    Science.gov (United States)

    Reed, Scott; Wood, Jon; Vazquez, Jose; Mignotte, Pierre-Yves; Privat, Benjamin

    2010-04-01

    Hull and harbor infrastructure inspections are frequently performed manually and involve quite a bit of risk and human and monetary resources. In any kind of threat and resource constrained environment, this involves unacceptable levels of risk and cost. Modern Remotely Operated Vehicles are highly refined machines that provide features and capabilities previously unavailable. Operations once carried out by divers can now be carried out more quickly, efficiently and safely by smart enabled ROVs. ROVs are rapidly deployable and capable of continuous, reliable operations in adverse conditions. They also provide a stable platform on which multiple sensors may be mounted and utilized to meet the harbor inspection problem. Automated Control software provides ROV's and their pilots with the capability to inspect complex, constrained environments such as those found in a harbor region. This application and the user interface allow the ROV to automatically conduct complex maneuvers relative to the area being inspected and relieves the training requirements and work load for the pilot, allowing he or she to focus on the primary task of survey, inspection and looking for possible threats (such as IEDs, Limpet Mines, signs of sabotage, etc). Real-time sensor processing tools can be integrated into the smart ROV solution to assist the operator. Automatic Target Recognition (ATR) algorithms are used to search through the sensor data collected by the ROV in real time. These algorithms provide immediate feedback on possible threats and notify the operator of regions that may require manual verification. Sensor data (sonar or video) is also mosaiced, providing the operator with real-time situational awareness and a coverage map of the hull or seafloor. Detected objects may also be placed in the context of the large scale characteristics of the hull (or bottom or pilings) and localized. Within the complex areas such as the harbor pier pilings and the running gear of the ship, real

  3. 33 CFR 334.1430 - Apra Inner Harbor, Island of Guam; restricted area.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Apra Inner Harbor, Island of Guam... THE ARMY, DEPARTMENT OF DEFENSE DANGER ZONE AND RESTRICTED AREA REGULATIONS § 334.1430 Apra Inner Harbor, Island of Guam; restricted area. (a) The restricted area. The waters within Apra Inner Harbor and...

  4. 33 CFR 165.1402 - Apra Outer Harbor, Guam-regulated navigation area.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Apra Outer Harbor, Guam-regulated....1402 Apra Outer Harbor, Guam—regulated navigation area. (a) The following is a regulated navigation area—The waters of the Pacific Ocean and Apra Outer Harbor enclosed by a line beginning at latitude 13...

  5. 77 FR 45239 - Amendment of Class E Airspace; Bar Harbor, ME

    Science.gov (United States)

    2012-07-31

    ...-1366; Airspace Docket No. 11-ANE-13] Amendment of Class E Airspace; Bar Harbor, ME AGENCY: Federal... area at Bar Harbor, ME, as the Surry Non-Directional Radio Beacon (NDB) has been decommissioned and new Standard Instrument Approach Procedures have been developed at Hancock County-Bar Harbor Airport. This...

  6. 77 FR 27666 - Proposed Amendment of Class E Airspace; Bar Harbor, ME

    Science.gov (United States)

    2012-05-11

    ...-1366; Airspace Docket No. 11-ANE-13] Proposed Amendment of Class E Airspace; Bar Harbor, ME AGENCY... action proposes to amend Class E Airspace at Bar Harbor, ME, as the Surry Non-Directional Radio Beacon... Hancock County-Bar Harbor Airport. This action would enhance the safety and airspace management of...

  7. 33 CFR 162.165 - Buffalo and Rochester Harbors, New York.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Buffalo and Rochester Harbors, New York. 162.165 Section 162.165 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND... and Rochester Harbors, New York. In Buffalo and Rochester Harbors, no vessel may exceed 6 miles per...

  8. 76 FR 32071 - Safety Zone; Conneaut Festival Fireworks, Conneaut Harbor, Conneaut, OH

    Science.gov (United States)

    2011-06-03

    ...-AA00 Safety Zone; Conneaut Festival Fireworks, Conneaut Harbor, Conneaut, OH AGENCY: Coast Guard, DHS... Conneaut Harbor, Conneaut, OH for the Conneaut Festival Fireworks. This zone is intended to restrict vessels from a portion of Conneaut Harbor, Conneaut, OH during the Conneaut Festival Fireworks on July 3...

  9. 78 FR 18479 - Drawbridge Operation Regulations; Inner Harbor Navigation Canal, New Orleans, LA

    Science.gov (United States)

    2013-03-27

    ... Operation Regulations; Inner Harbor Navigation Canal, New Orleans, LA AGENCY: Coast Guard, DHS. ACTION... across the Inner Harbor Navigation Canal, mile 4.6, at New Orleans, Louisiana. This deviation is... Seabrook Highway crossing the Inner Harbor Navigation Canal, mile 4.6, in New Orleans, Louisiana. The...

  10. 76 FR 38153 - California State Nonroad Engine Pollution Control Standards; Commercial Harbor Craft Regulations...

    Science.gov (United States)

    2011-06-29

    ... Standards; Commercial Harbor Craft Regulations; Opportunity for Public Hearing and Comment AGENCY... engines on commercial harbor craft. CARB has requested that EPA issue a new authorization under [email protected] . SUPPLEMENTARY INFORMATION: I. California's Commercial Harbor Craft Regulations In a...

  11. Probing the emitter site of Renilla luciferase using small organic molecules; an attempt to understand the molecular architecture of the emitter site.

    Science.gov (United States)

    Salehi, Farajollah; Emamzadeh, Rahman; Nazari, Mahboobeh; Rasa, Seyed Mohammad Mahdi

    2016-12-01

    Renilla luciferase is a sensitive enzyme and has wide applications in biotechnology such as drug screening. Previous studies have tried to show the catalytic residues, nevertheless, the accurate architecture and molecular behavior of its emitter site remains uncharacterized. In this study, the activity of Renilla luciferase, in the presence of two small organic molecules including dimethyl sulfoxide (DMSO) and isopropanol was considered and the structure was studied by circular dichroism (CD) and fluorescence spectroscopy. Moreover, the interaction of small organic molecules with the Renilla luciferase was studied using molecular dynamics simulations. Kinetics studies showed that at low concentration of DMSO (16.6-66mM) and isopropanol (19.3-76mM) the K m changed and a competitive inhibition pattern was observed. Moreover, spectroscopy studies reveled that the changes of activity of Renilla luciferase in the presence of low concentrations of small organic molecules was not associated with structural collapse or severe changes in the enzyme conformation. Molecular dynamics simulations indicated that DMSO and isopropanol, as probing molecules, were both able to bind to the emitter site and remained with the residues of the emitter site. Based on the probing data, the architecture of the emitter site in the "non-binding" model was proposed. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Surprise: Dwarf Galaxy Harbors Supermassive Black Hole

    Science.gov (United States)

    2011-01-01

    with the Hubble Space Telescope. They found a region near the center of the galaxy that strongly emits radio waves with characteristics of those emitted by super-fast "jets" of material spewed outward from areas close to a black hole. They then searched images from the Chandra X-Ray Observatory that showed this same, radio-bright region to be strongly emitting energetic X-rays. This combination, they said, indicates an active, black-hole-powered, galactic nucleus. "Not many dwarf galaxies are known to have massive black holes," Sivakoff said. While central black holes of roughly the same mass as the one in Henize 2-10 have been found in other galaxies, those galaxies all have much more regular shapes. Henize 2-10 differs not only in its irregular shape and small size but also in its furious star formation, concentrated in numerous, very dense "super star clusters." "This galaxy probably resembles those in the very young Universe, when galaxies were just starting to form and were colliding frequently. All its properties, including the supermassive black hole, are giving us important new clues about how these black holes and galaxies formed at that time," Johnson said. The astronomers reported their findings in the January 9 online edition of Nature, and at the American Astronomical Society's meeting in Seattle, WA.

  13. Hydrocarbon pollutants shape bacterial community assembly of harbor sediments

    KAUST Repository

    Barbato, Marta

    2016-02-02

    Petroleum pollution results in co-contamination by different classes of molecules, entailing the occurrence of marine sediments difficult to remediate, as in the case of the Ancona harbor (Mediterranean Sea, Italy). Autochthonous bioaugmentation (ABA), by exploiting the indigenous microbes of the environment to be treated, could represent a successful bioremediation strategy. In this perspective we aimed to i) identify the main drivers of the bacterial communities\\' richness in the sediments, ii) establish enrichment cultures with different hydrocarbon pollutants evaluating their effects on the bacterial communities\\' composition, and iii) obtain a collection of hydrocarbon degrading bacteria potentially exploitable in ABA. The correlation between the selection of different specialized bacterial populations and the type of pollutants was demonstrated by culture-independent analyses, and by establishing a collection of bacteria with different hydrocarbon degradation traits. Our observations indicate that pollution dictates the diversity of sediment bacterial communities and shapes the ABA potential in harbor sediments.

  14. Army Engineer Divers: First In Port-Au-Prince Harbor

    Science.gov (United States)

    2010-12-01

    to the pile once the concrete was poured. This step was com- pleted in a few minutes for each pile. Last, a prefabricated wooden form was emplaced...September-December 201010 Engineer This prefabricated wooden form was emplaced around a rebar cage at the top of damaged piles to hold concrete until it...the mainland into the harbor. Waste from tugboats and sewage from the mainland compounded ecological hazards. The only alternative for the Army and

  15. An Evaluation of the Acquisition Streamlining Methods at the Fleet and Industrial Supply Center Pearl Harbor Hawaii

    National Research Council Canada - National Science Library

    Henry, Mark

    1999-01-01

    ...) Pearl Harbor's implementation of acquisition streamlining initiatives and recommends viable methods of streamlining the acquisition process at FISC Pearl Harbor and other Naval Supply Systems Command...

  16. Towards an ethics safe harbor for global biomedical research

    Science.gov (United States)

    Dove, Edward S.; Knoppers, Bartha M.; Zawati, Ma'n H.

    2014-01-01

    Although increasingly global, data-driven genomics and other ‘omics’-focused research hold great promise for health discoveries, current research ethics review systems around the world challenge potential improvements in human health from such research. To overcome this challenge, we propose a ‘Safe Harbor Framework for International Ethics Equivalency’ that facilitates the harmonization of ethics review of specific types of data-driven international research projects while respecting globally transposable research ethics norms and principles. The Safe Harbor would consist in part of an agency supporting an International Federation for Ethics Review (IFER), formed by a voluntary compact among countries, granting agencies, philanthropies, institutions, and healthcare, patient advocacy, and research organizations. IFER would be both a central ethics review body, and also a forum for review and follow-up of policies concerning ethics norms for international research projects. It would be built on five principle elements: (1) registration, (2) compliance review, (3) recognition, (4) monitoring and enforcement, and (5) public participation. The Safe Harbor would create many benefits for researchers, countries, and the general public, and may eventually have application beyond (gen)omics to other areas of biomedical research that increasingly engage in secondary use of data and present only negligible risks. PMID:27774154

  17. Pearl Harbor: lessons for the dam safety community

    Energy Technology Data Exchange (ETDEWEB)

    Martin, T.E. [AMEC Earth and Environmental Ltd., Burnaby, BC (Canada)

    2001-10-01

    Every good dam safety program must be based on surveillance and emergency response planning. The same principles apply to the gathering of information for military intelligence and the planning of defence tactics. Lessons learned from failure have spurred the advancement of dam engineering. Dam safety experts can benefit from the inadequacies encountered by the military community, with the most famous occurring on December 7, 1941 in Pearl Harbor. Both intelligence gathering and contingency response planning failed miserably. The data was not properly disseminated, interpreted, analysed. The proper response to the situation was not initiated. Human error and failure to communicate are the two main reasons that explain the debacle. The inquiries into the tragedy at Pearl Harbor provided valuable lessons, related to individual and organizational failures, which the authors shared in this presentation. The relevance to dam safety was made. All Federal Bureau of Investigation (FBI) agents must read the lessons drawn from Pearl Harbor, as they have responsibility for dam safety. 4 refs.

  18. Movements and foraging effort of Steller's Eiders and Harlequin Ducks wintering near Dutch Harbor, Alaska

    Science.gov (United States)

    Reed, J.A.; Flint, Paul L.

    2007-01-01

    We studied the movements and foraging effort of radio-marked Steller's Eiders (Polysticta stelleri) and Harlequin Ducks (Histrionicus histrionicus) to evaluate habitat quality in an area impacted by industrial activity near Dutch Harbor, Alaska. Foraging effort was relatively low, with Steller's Eiders foraging only 2.7 ± 0.6 (SE) hours per day and Harlequin Ducks 4.1 ± 0.5 hours per day. Low-foraging effort during periods of high-energetic demand generally suggests high food availability, and high food availability frequently corresponds with reductions in home range size. However, the winter ranges of Harlequin Ducks did not appear to be smaller than usual, with the mean range size in our study (5.5 ± 1.1 km2) similar to that reported by previous investigators. The mean size of the winter ranges of Steller's Eiders was similar (5.1 ± 1.3 km2), but no comparable estimates are available. Eutrophication of the waters near Dutch Harbor caused by seafood processing and municipal sewage effluent may have increased populations of the invertebrate prey of these sea ducks and contributed to their low-foraging effort. The threat of predation by Bald Eagles (Haliaeetus leucocephalus) that winter near Dutch Harbor may cause Steller's Eiders and Harlequin Ducks to move further offshore when not foraging, contributing to an increase in range sizes. Thus, the movement patterns and foraging behavior of these ducks likely represent a balance between the cost and benefits of wintering in a human-influenced environment.

  19. A solitary hyperfunctioning thyroid nodule harboring thyroid carcinoma: review of the literature.

    Science.gov (United States)

    Mirfakhraee, Sasan; Mathews, Dana; Peng, Lan; Woodruff, Stacey; Zigman, Jeffrey M

    2013-05-04

    Hyperfunctioning nodules of the thyroid are thought to only rarely harbor thyroid cancer, and thus are infrequently biopsied. Here, we present the case of a patient with a hyperfunctioning thyroid nodule harboring thyroid carcinoma and, using MEDLINE literature searches, set out to determine the prevalence of and characteristics of malignant "hot" nodules as a group. Historical, biochemical and radiologic characteristics of the case subjects and their nodules were compared to those in cases of benign hyperfunctioning nodules. A literature review of surgical patients with solitary hyperfunctioning thyroid nodules managed by thyroid resection revealed an estimated 3.1% prevalence of malignancy. A separate literature search uncovered 76 cases of reported malignant hot thyroid nodules, besides the present case. Of these, 78% were female and mean age at time of diagnosis was 47 years. Mean nodule size was 4.13 ± 1.68 cm. Laboratory assessment revealed T3 elevation in 76.5%, T4 elevation in 51.9%, and subclinical hyperthyroidism in 13% of patients. Histological diagnosis was papillary thyroid carcinoma (PTC) in 57.1%, follicular thyroid carcinoma (FTC) in 36.4%, and Hurthle cell carcinoma in 7.8% of patients. Thus, hot thyroid nodules harbor a low but non-trivial rate of malignancy. Compared to individuals with benign hyperfunctioning thyroid nodules, those with malignant hyperfunctioning nodules are younger and more predominantly female. Also, FTC and Hurthle cell carcinoma are found more frequently in hot nodules than in general. We were unable to find any specific characteristics that could be used to distinguish between malignant and benign hot nodules.

  20. Development of Site-Specific Water Quality Criteria for the Arpa Harbor Wastewater Treatment Plant in Tipalao Bay, Guam

    Science.gov (United States)

    2016-07-01

    flats, estuaries, and coastal creeks. It is usually found within tidally influenced waters , but it may enter freshwater, ascending about 10...based on a dissolved metal concentration endpoint in the receiving body of water . In other words , the translator supports estimations of the fraction of...TECHNICAL REPORT 2068 July 2016 Development of Site-Specific Water Quality Criteria for the Arpa Harbor Wastewater Treatment Plant

  1. Effects of Harbor Modification on Crescent City, California's Tsunami Vulnerability

    Science.gov (United States)

    Dengler, Lori; Uslu, Burak

    2011-06-01

    More damaging tsunamis have impacted Crescent City, California in historic times than any other location on the West Coast of the USA. Crescent City's harbor has undergone significant modification since the early 20th century, including construction of several breakwaters, dredging, and a 200 × 300 m2 small boat basin. In 2006, a M w 8.3 earthquake in the Kuril Islands generated a moderate Pacific-wide tsunami. Crescent City recorded the highest amplitudes of any tide gauge in the Pacific and was the only location to experience structural damage. Strong currents damaged docks and boats within the small boat basin, causing more than US 20 million in damage and replacement costs. We examine how modifications to Crescent City's harbor may have affected its vulnerability to moderate tsunamis such as the 2006 event. A bathymetric grid of the basin was constructed based on US Army Corps of Engineers soundings in 1964 and 1965 before the construction of the small boat basin. The method of splitting tsunamis was used to estimate tsunami water heights and current velocities at several locations in the harbor using both the 1964-1965 grid and the 2006 bathymetric grid for the 2006 Kuril event and a similar-sized source along the Sanriku coast of Japan. Model velocity outputs are compared for the two different bathymetries at the tide gauge location and at six additional computational sites in the harbor. The largest difference between the two grids is at the small boat basin entrance, where the 2006 bathymetry produces currents over three times the strength of the currents produced by the 1965 bathymetry. Peak currents from a Sanriku event are comparable to those produced by the 2006 event, and within the boat basin may have been higher. The modifications of the harbor, and in particular the addition of the small boat basin, appear to have contributed to the high current velocities and resulting damage in 2006 and help to explain why the 1933 M w 8.4-8.7 Sanriku tsunami

  2. Grays Harbor and Chehalis River Improvements to Navigation Environmental Studies. Wildlife Studies at Proposed Disposal Sites in Grays Harbor, Washington,

    Science.gov (United States)

    1982-01-01

    sltand. T 𔃼~P i’ W 210 three times VtwCerI November IOC’C -nd ~co l.Etls ~ ec!,!zervc-o betxwe H -gF 12 Th -ind hl rway u- 7Plie Sicuobh. E. Cumin -s 1... stress imposed by dredge dsosal ;ictivities on these species. It is difficult to rredict the effects of establishing a salt marsh in Grays Harbor on

  3. Production of infectious chimeric hepatitis C virus genotype 2b harboring minimal regions of JFH-1.

    Science.gov (United States)

    Murayama, Asako; Kato, Takanobu; Akazawa, Daisuke; Sugiyama, Nao; Date, Tomoko; Masaki, Takahiro; Nakamoto, Shingo; Tanaka, Yasuhito; Mizokami, Masashi; Yokosuka, Osamu; Nomoto, Akio; Wakita, Takaji

    2012-02-01

    To establish a cell culture system for chimeric hepatitis C virus (HCV) genotype 2b, we prepared a chimeric construct harboring the 5' untranslated region (UTR) to the E2 region of the MA strain (genotype 2b) and the region of p7 to the 3' UTR of the JFH-1 strain (genotype 2a). This chimeric RNA (MA/JFH-1.1) replicated and produced infectious virus in Huh7.5.1 cells. Replacement of the 5' UTR of this chimera with that from JFH-1 (MA/JFH-1.2) enhanced virus production, but infectivity remained low. In a long-term follow-up study, we identified a cell culture-adaptive mutation in the core region (R167G) and found that it enhanced virus assembly. We previously reported that the NS3 helicase (N3H) and the region of NS5B to 3' X (N5BX) of JFH-1 enabled replication of the J6CF strain (genotype 2a), which could not replicate in cells. To reduce JFH-1 content in MA/JFH-1.2, we produced a chimeric viral genome for MA harboring the N3H and N5BX regions of JFH-1, combined with a JFH-1 5' UTR replacement and the R167G mutation (MA/N3H+N5BX-JFH1/R167G). This chimeric RNA replicated efficiently, but virus production was low. After the introduction of four additional cell culture-adaptive mutations, MA/N3H+N5BX-JFH1/5am produced infectious virus efficiently. Using this chimeric virus harboring minimal regions of JFH-1, we analyzed interferon sensitivity and found that this chimeric virus was more sensitive to interferon than JFH-1 and another chimeric virus containing more regions from JFH-1 (MA/JFH-1.2/R167G). In conclusion, we established an HCV genotype 2b cell culture system using a chimeric genome harboring minimal regions of JFH-1. This cell culture system may be useful for characterizing genotype 2b viruses and developing antiviral strategies.

  4. Specific detection of peste des petits ruminants virus antibodies in sheep and goat sera by the luciferase

    International Nuclear Information System (INIS)

    Berguido, F.J.; Bodjo, S.C.; Loitsch, A.; Diallo, A.

    2016-01-01

    Full text: Peste des petits ruminants (PPR) is a contagious and often fatal transboundary animal disease affecting mostly sheep, goats and wild small ruminants. This disease is endemic in most of Africa, the Middle, Near East, and large parts of Asia. The casual agent is peste des petits ruminants virus (PPRV), which belongs to the genus Morbilivirus in the family Paramyxoviridae. This genus also includes measles virus (MV), canine distemper virus (CDV) and rinderpest virus (RPV). All are closely related viruses with serological cross reactivity. In this study, we have developed a Luciferase Immunoprecipitation System (LIPS) for the rapid detection of antibodies against PPRV in serum samples and for specific differentiation from antibodies against RPV. PPR and rinderpest (RP) serum samples were assayed by PPR-LIPS and two commercially available PPR cELISA tests. The PPR-LIPS showed high sensitivity and specificity for the samples tested and showed no cross reactivity with RPV unlike the commercial PPR cELISA tests which did not cross react with RPV. Based on the results shown in this study, PPR-LIPS is presented as a good candidate for the specific serosurveillance of PPR. (author)

  5. High-throughput screening of effective siRNAs using luciferase-linked chimeric mRNA.

    Directory of Open Access Journals (Sweden)

    Shen Pang

    Full Text Available The use of siRNAs to knock down gene expression can potentially be an approach to treat various diseases. To avoid siRNA toxicity the less transcriptionally active H1 pol III promoter, rather than the U6 promoter, was proposed for siRNA expression. To identify highly efficacious siRNA sequences, extensive screening is required, since current computer programs may not render ideal results. Here, we used CCR5 gene silencing as a model to investigate a rapid and efficient screening approach. We constructed a chimeric luciferase-CCR5 gene for high-throughput screening of siRNA libraries. After screening approximately 900 shRNA clones, 12 siRNA sequences were identified. Sequence analysis demonstrated that most (11 of the 12 sequences of these siRNAs did not match those identified by available siRNA prediction algorithms. Significant inhibition of CCR5 in a T-lymphocyte cell line and primary T cells by these identified siRNAs was confirmed using the siRNA lentiviral vectors to infect these cells. The inhibition of CCR5 expression significantly protected cells from R5 HIV-1JRCSF infection. These results indicated that the high-throughput screening method allows efficient identification of siRNA sequences to inhibit the target genes at low levels of expression.

  6. Experimental evidence for the physiological role of bacterial luciferase in the protection of cells against oxidative stress.

    Science.gov (United States)

    Szpilewska, Hanna; Czyz, Agata; Wegrzyn, Grzegorz

    2003-11-01

    The origin and function of bioluminescence was considered a problematic question of the Charles Darwin theory. Early evolution of bacterial luminescence and its current physiological importance seem to be especially mysterious. Recently, it was proposed that stimulation of DNA repair may be a physiological role for production of light by bacterial cells. On the other hand, it was also proposed that primary role of luminescent systems could be detoxification of the deleterious oxygen derivatives. Although some previous results might suggest that this hypothesis can be correct, until now experimental evidence for such a mechanism operating in bacterial cells and having physiological importance was generally lacking. Here we demonstrate that in the presence of various oxidants (hydrogen peroxide, cumene hydroperoxide, t-butyl hydroperoxide, and ferrous ions) at certain concentrations in the culture medium, growth of Vibrio harveyi mutants luxA and luxB, but not of the mutant luxD, is severely impaired relative to wild-type bacteria. This deleterious effect of oxidants on the mutants luxA and luxB could be significantly reduced by addition of the antioxidants A-TEMPO or 40H-TEMPO. We conclude that bacterial luciferase may indeed play a physiological role in the protection of cells against oxidative stress.

  7. ATPase inhibitor based luciferase assay for prolonged and enhanced ATP pool measurement as an efficient fish freshness indicator.

    Science.gov (United States)

    Ranjan, Rajeev; Priyanka, B S; Thakur, M S

    2014-07-01

    The nucleotide degradation pathway in somatic cells leads to the accumulation of products such as hypoxanthine and inosine, which are commonly used as fish and meat freshness indicators. Assays based on these molecules cannot differentiate the postmortem time over a short period of time (5-10 h). Further, quantification of these degradation products is cumbersome, costly and time-consuming. For the proposed assay, optimal concentrations of 30 and 2 mM, respectively, for the ATPase inhibitors sodium orthovanadate and EDTA were found. Further, it was observed that a firefly luciferase based assay could enhance the sensitivity levels up to 165-fold at 30 °C. In addition, it was observed that the sensitivity for ATP assay was enhanced up to 60-fold even after 12 h. The limit of detection for the ATP assay was 1 pM, unlike other conventional methods, which are sensitive only up to micromolar levels. Moreover, as little as 0.044 g fish fillet was required for the assay, and no time-consuming sample preparation was necessary. Luminescence of prolonged duration was observed in harvested fish kept at -20 °C in comparison with fish kept at 4 and 30 °C, which reflects the shelf life of fish preserved at lower temperatures.

  8. An in vitro test to screen skin sensitizers using a stable THP-1-derived IL-8 reporter cell line, THP-G8.

    Science.gov (United States)

    Takahashi, Toshiya; Kimura, Yutaka; Saito, Rumiko; Nakajima, Yoshihiro; Ohmiya, Yoshihiro; Yamasaki, Kenshi; Aiba, Setsuya

    2011-12-01

    Several studies have suggested that interleukin (IL)-8 can serve as a biomarker for discrimination of skin sensitizers from nonsensitizers. We established a stable THP-1-derived IL-8 reporter cell line, THP-G8, which harbors SLO and SLR luciferase genes under the control of IL-8 and glyceraldehyde 3-phosphate dehydrogenase promoters, respectively. After 6 h treatment with chemicals, normalized SLO luciferase activity (nSLO-LA) was calculated by dividing SLO-LA by SLR-LA, and the fold induction of nSLO-LA (FInSLO-LA) was calculated by dividing nSLO-LA of chemically treated cells by that of nontreated cells. The nSLO-LA of THP-G8 cells increased in response to lipopolysaccharide (LPS) and several sensitizers. The FInSLO-LA in THP-G8 cells induced by LPS or sensitizers positively correlated with their induction of IL-8 messenger RNA in THP-1 cells. The nSLO-LA value of THP-G8 cells was significantly increased (FInSLO-LA ≥ 1.4) by 13 of the 15 sensitizers as well as by 5 of the 7 nonsensitizers. Interestingly, pretreatment with N-acetylcysteine suppressed the increase in FInSLO-LA induced by all sensitizers (inhibition index (II) ≤ 0.8) but did not suppress that induced by most of the nonsensitizers. We then evaluated the performance of this assay using values of FInSLO-LA ≥ 1.4 and II ≤ 0.8 in at least two of three independent experiments as the criteria of a sensitizer, which resulted in test accuracies of 82% for the 22 chemicals used and of 88% for the chemicals proposed by European Center for the Validation of Alternative Methods. This newly developed assay is a candidate replacement for animal tests of skin sensitization because of its accuracy, convenience, and high throughput performance.

  9. Human case of bacteremia caused by Streptococcus canis sequence type 9 harboring the scm gene.

    Science.gov (United States)

    Taniyama, Daisuke; Abe, Yoshihiko; Sakai, Tetsuya; Kikuchi, Takahide; Takahashi, Takashi

    2017-01-01

    Streptococcus canis (Sc) is a zoonotic pathogen that is transferred mainly from companion animals to humans. One of the major virulence factors in Sc is the M-like protein encoded by the scm gene, which is involved in anti-phagocytic activities, as well as the recruitment of plasminogen to the bacterial surface in cooperation with enolase, and the consequent enhancement of bacterial transmigration and survival. This is the first reported human case of uncomplicated bacteremia following a dog bite, caused by Streptococcus canis harboring the scm gene. The similarity of the 16S rRNA from the infecting species to that of the Sc type strain, as well as the amplification of the species-specific cfg gene, encoding a co-hemolysin, was used to confirm the species identity. Furthermore, the isolate was confirmed as sequence type 9. The partial scm gene sequence harbored by the isolate was closely related to those of other two Sc strains. While this isolate did not possess the erm (A), erm (B), or mef (A), macrolide/lincosamide resistance genes, it was not susceptible to azithromycin: its susceptibility was intermediate. Even though human Sc bacteremia is rare, clinicians should be aware of this microorganism, as well as Pasteurella sp., Prevotella sp., and Capnocytophaga sp., when examining and treating patients with fever who maintain close contact with companion animals.

  10. A case of lung adenocarcinoma harboring EGFR mutation and EML4-ALK fusion gene

    International Nuclear Information System (INIS)

    Tanaka, Hisashi; Hayashi, Akihito; Morimoto, Takeshi; Taima, Kageaki; Tanaka, Yoshihito; Shimada, Michiko; Kurose, Akira; Takanashi, Shingo; Okumura, Ken

    2012-01-01

    Lung cancer is the leading cause of cancer-related death worldwide. Epidermal growth factor receptor (EGFR) - tyrosine kinase inhibitor (TKI) is used for the patients with EGFR-mutant lung cancer. Recently, phase III studies in the patients with EGFR-mutant demonstrated that EGFR-TKI monotherapy improved progression-free survival compared with platinum-doublet chemotherapy. The echinoderm microtubule-associated protein-like 4 (EML4) - anaplastic lymphoma kinase (ALK) fusion oncogene represents one of the newest molecular targets in non-small cell lung cancer (NSCLC). Patients who harbor EML4-ALK fusions have been associated with a lack of EGFR or KRAS mutations. We report a 39-year-old patient diagnosed as adenocarcinoma harboring EGFR mutation and EML4-ALK fusion gene. We treated this patient with erlotinib as the third line therapy, but no clinical benefit was obtained. We experienced a rare case with EGFR mutation and EML4-ALK. Any clinical benefit using EGFR-TKI was not obtained in our case. The therapeutic choice for the patients with more than one driver mutations is unclear. We needs further understanding of the lung cancer molecular biology and the biomarker infomation

  11. Caffeine in Boston Harbor past and present, assessing its utility as a tracer of wastewater contamination in an urban estuary

    Science.gov (United States)

    Sites throughout Boston Harbor were analyzed for caffeine to assess its utility as a tracer in identifying sources of sanitary wastewater. Caffeine ranged from 15 ng/L in the outer harbor to a high of 185 ng/L in the inner harbor. Inner harbor concentrations were a result of comb...

  12. Heat from harbor to city; Warmte van haven naar stad

    Energy Technology Data Exchange (ETDEWEB)

    Roggen, M. (ed.)

    2004-09-01

    The Rotterdam harbor area is perfect for an industrial heat distribution network. Numerous major heat producers and many potential consumers of all sizes group together in this industrial area. The new Heat Company ('Warmtebedrijf') is making a detailed business plan to build a heat distribution system. [Dutch] Als er een regio is waar een industrieel warmtenet voor de hand ligt, is het het Rotterdamse havengebied wel. Tal van grote warmteproducenten en een woud aan kleinere en grotere afnemers zijn praktisch buren. Om handen en voeten te geven aan dit op papier mooie idee, stelt het kersverse Warmtebedrijf io een gedetailleerd businessplan op.

  13. Hydraulic modeling of stream channels and structures in Harbor and Crow Hollow Brooks, Meriden, Connecticut

    Science.gov (United States)

    Weiss, Lawrence A.; Sears, Michael P.; Cervione, Michael A.

    1994-01-01

    Effects of urbanization have increased the frequency and size of floods along certain reaches of Harbor Brook and Crow Hollow Brook in Meriden, Conn. A floodprofile-modeling study was conducted to model the effects of selected channel and structural modifications on flood elevations and inundated areas. The study covered the reach of Harbor Brook downstream from Interstate 691 and the reach of Crow Hollow Brook downstream from Johnson Avenue. Proposed modifications, which include changes to bank heights, channel geometry, structural geometry, and streambed armoring on Harbor Brook and changes to bank heights on Crow Hollow Brook, significantly lower flood elevations. Results of the modeling indicate a significant reduction of flood elevations for the 10-year, 25-year, 35-year, 50-year, and 100-year flood frequencies using proposed modifications to (1 ) bank heights between Harbor Brook Towers and Interstate 691 on Harbor Brook, and between Centennial Avenue and Johnson Avenue on Crow Hollow Brook; (2) channel geometry between Coe Avenue and Interstate 69 1 on Harbor Brook; (3) bridge and culvert opening geometry between Harbor Brook Towers and Interstate 691 on Harbor Brook; and (4) channel streambed armoring between Harbor Brook Towers and Interstate 691 on Harbor Brook. The proposed modifications were developed without consideration of cost-benefit ratios.

  14. A Luciferase-Expressing Leishmania braziliensis Line That Leads to Sustained Skin Lesions in BALB/c Mice and Allows Monitoring of Miltefosine Treatment Outcome.

    Directory of Open Access Journals (Sweden)

    Adriano C Coelho

    2016-05-01

    Full Text Available Leishmania braziliensis is the most prevalent species isolated from patients displaying cutaneous and muco-cutaneous leishmaniasis in South America. However, there are difficulties for studying L. braziliensis pathogenesis or response to chemotherapy in vivo due to the natural resistance of most mouse strains to infection with these parasites. The aim of this work was to develop an experimental set up that could be used to assess drug efficacy against L. braziliensis. The model was tested using miltefosine.A L. braziliensis line, originally isolated from a cutaneous leishmaniasis patient, was passaged repeatedly in laboratory rodents and further genetically manipulated to express luciferase. Once collected from a culture of parasites freshly transformed from amastigotes, 106 wild type or luciferase-expressing stationary phase promastigotes were inoculated subcutaneously in young BALB/c mice or golden hamsters. In both groups, sustained cutaneous lesions developed at the site of inoculation, no spontaneous self- healing being observed 4 months post-inoculation, if left untreated. Compared to the wild type line features, no difference was noted for the luciferase-transgenic line. Infected animals were treated with 5 or 15 mg/kg/day miltefosine orally for 15 days. At the end of treatment, lesions had regressed and parasites were not detected. However, relapses were observed in animals treated with both doses of miltefosine.Here we described experimental settings for a late-healing model of cutaneous leishmaniasis upon inoculation of a luciferase-expressing L. braziliensis line that can be applied to drug development projects. These settings allowed the monitoring of the transient efficacy of a short-term miltefosine administration.

  15. Reporter gene assay for the quantification of the activity and neutralizing antibody response to TNFα antagonists

    DEFF Research Database (Denmark)

    Lallemand, Christophe; Kavrochorianou, Nadia; Steenholdt, Casper

    2011-01-01

    A cell-based assay has been developed for the quantification of the activity of TNFα antagonists based on human erythroleukemic K562 cells transfected with a NFκB regulated firefly luciferase reporter-gene construct. Both drug activity and anti-drug neutralizing antibodies can be quantified...... with a high degree of precision within 2h, and without interference from cytokines and other factors known to activate NFκB. The assay cells also contain the Renilla luciferase reporter gene under the control of a constitutive promoter that allows TNFα-induced firefly luciferase activity to be normalized...... relative to Renilla luciferase expression. Thus, results are independent of cell number or differences in cell viability, resulting in intra and inter assay coefficients of variation of 10% or less. Normalization of results relative to the expression of an internal standard also provides a means...

  16. Use of nuclear receptor luciferase-based bioassays to detect endocrine active chemicals in a biosolids-biochar amended soil.

    Science.gov (United States)

    Anderson, Carolyn G; Joshi, Geetika; Bair, Daniel A; Oriol, Charlotte; He, Guochun; Parikh, Sanjai J; Denison, Michael S; Scow, Kate M

    2017-08-01

    Biosolids are a potentially valuable source of carbon and nutrients for agricultural soils; however, potential unintended impacts on human health and the environment must be considered. Virtually all biosolids contain trace amounts endocrine-disrupting chemicals derived from human use of pharmaceuticals and personal care products (PPCPs). One potential way to reduce the bioavailability of PPCPs is to co-apply biosolids with biochar to soil, because biochar's chemical (e.g., aromaticity) and physical properties (e.g., surface area) give it a high affinity to bind many organic chemicals in the environment. We developed a soil-specific extraction method and utilized a luciferase-based bioassay (CALUX) to detect endocrine active chemicals in a biosolids-biochar co-amendment soil greenhouse study. Both biochar (walnut shell, 900 °C) and biosolids had positive impacts on carrot and lettuce biomass accumulation over our study period. However, the walnut shell biochar stimulated aryl hydrocarbon receptor activity, suggesting the presence of potential endocrine active chemicals in the biochar. Since the biochar rate tested (100 t ha -1 ) is above the average agronomic rate (10-20 t ha -1 ), endocrine effects would not be expected in most environmental applications. The effect of high temperature biochars on endocrine system pathways must be explored further, using both quantitative analytical tools to identify potential endocrine active chemicals and highly sensitive bioanalytical assays such as CALUX to measure the resulting biological activity of such compounds. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Bluff evolution along coastal drumlins: Boston Harbor Islands, Massachusetts

    Science.gov (United States)

    Himmelstoss, E.A.; FitzGerald, D.M.; Rosen, P.S.; Allen, J.R.

    2006-01-01

    A series of partially drowned drumlins forms the backbone of the inner islands within Boston Harbor. The shoreline of these rounded glacial deposits is composed of actively retreating bluffs formed by continual wave attack. Comparisons of bluffs reveal variability in their height and lateral extent, as well as in the dominant mechanism causing their retreat. Two processes are responsible for bluff erosion and yield distinct bluff morphologies: (1) wave attack undercuts the bluff and causes episodic slumping, yielding planar bluff slopes, and (2) subaerial processes such as rainfall create irregular slopes characterized by rills and gullies. We propose a model of drumlin bluff evolution that is based on processes of erosion and physical characteristics such as bluff height, slope morphology, and the orientation of the bluff with respect to the long axis of the drumlin and its topographic crest. The four phases of drumlin bluff evolution consist of (1) initial formation of bluff, with retreat dominated by wave notching and slumping processes; (2) rill and gully development as bluff heights exceed 10 m and slumped sediment at bluff base inhibits wave attack; (3) return of wave notching and slumping as bluff heights decrease; and (4) final development of boulder retreat lag as last remnants of drumlin are eroded by wave action. These phases capture the important physical processes of drumlin evolution in Boston Harbor and could apply to other eroding coastal drumlin deposits.

  18. New world bats harbor diverse influenza A viruses.

    Directory of Open Access Journals (Sweden)

    Suxiang Tong

    Full Text Available Aquatic birds harbor diverse influenza A viruses and are a major viral reservoir in nature. The recent discovery of influenza viruses of a new H17N10 subtype in Central American fruit bats suggests that other New World species may similarly carry divergent influenza viruses. Using consensus degenerate RT-PCR, we identified a novel influenza A virus, designated as H18N11, in a flat-faced fruit bat (Artibeus planirostris from Peru. Serologic studies with the recombinant H18 protein indicated that several Peruvian bat species were infected by this virus. Phylogenetic analyses demonstrate that, in some gene segments, New World bats harbor more influenza virus genetic diversity than all other mammalian and avian species combined, indicative of a long-standing host-virus association. Structural and functional analyses of the hemagglutinin and neuraminidase indicate that sialic acid is not a ligand for virus attachment nor a substrate for release, suggesting a unique mode of influenza A virus attachment and activation of membrane fusion for entry into host cells. Taken together, these findings indicate that bats constitute a potentially important and likely ancient reservoir for a diverse pool of influenza viruses.

  19. Grace Under Fire: The Army Nurses of Pearl Harbor, 1941.

    Science.gov (United States)

    Milbrath, Gwyneth R

    2016-01-01

    Much has been written about the military events of December 7, 1941; however, little has been documented about the nurses' work and experience at Pearl Harbor, Hawaii. The aerial assault on Pearl Harbor was the first time in US history that Army nurses had been on the front line of battle. Nurses quickly triaged and stabilized those who could be saved, and provided compassion and comfort to those who were dying, in an environment where the nurses were unsure of their own survival. Traditional historical methods and a social history framework were used in this investigation. Primary sources included oral histories from the US Army Medical Department Center of History and Heritage and the State of Hawaii's website, Hawaii Aviation. Secondary sources included published books, newspaper articles, military websites, and history texts. Due to the limited bed capacity, Hickam Field Hospital converted to an evacuation hospital. Nurses, physicians, and medical corpsman triaged, stabilized, and transported those likely to survive, while staging the dead behind the building. The emergency room at Tripler Hospital was quickly flooded with patients from the battlefield, but the staff was able to sort patients appropriately to the wards, to the operating room, or provide comfort care as they died. At Schofield Hospital, collaboration between tireless doctors, nurses, and corpsmen was key to providing life-saving surgery and care.

  20. The historical significance of anaesthesia events at Pearl Harbor.

    Science.gov (United States)

    Crowhurst, Ja

    2014-07-01

    Up to the end of World War II, less than 10% of the general anaesthetics administered was with intravenous barbiturates. The remaining 90% of anaesthetics given in the USA were with diethyl ether. In the United Kingdom and elsewhere, chloroform was also popular. Diethyl ether administration was a relatively safe and simple procedure, often delegated to nurses or junior doctors with little or no specific training in anaesthesia. During the Japanese attack on the US bases at Pearl Harbor, with reduced stocks of diethyl ether available, intravenous Sodium Pentothal(®), a most 'sophisticated and complex' drug, was used with devastating effects in many of those hypovolaemic, anaemic and septic patients. The hazards of spinal anaesthesia too were realised very quickly. These effects were compounded by the dearth of trained anaesthetists. This paper presents the significance of the anaesthesia tragedies at Pearl Harbor, and the discovery in the next few years of many other superior drugs that caused medical and other health professionals to realise that anaesthesia needed to be a specialist medical discipline in its own right. Specialist recognition, aided by the foundation of the National Health Service in the UK, the establishment of Faculties of Anaesthesia and appropriate training in pharmacology, physiology and other sciences soon followed. Modern anaesthesiology, as we understand it today, was born and a century or more of ether anaesthesia finally ceased.

  1. Relationship of the luminous bacterial symbiont of the Caribbean flashlight fish, Kryptophanaron alfredi (family Anomalopidae) to other luminous bacteria based on bacterial luciferase (luxA) genes.

    Science.gov (United States)

    Haygood, M G

    1990-01-01

    Flashlight fishes (family Anomalopidae) have light organs that contain luminous bacterial symbionts. Although the symbionts have not yet been successfully cultured, the luciferase genes have been cloned directly from the light organ of the Caribbean species, Kryptophanaron alfredi. The goal of this project was to evaluate the relationship of the symbiont to free-living luminous bacteria by comparison of genes coding for bacterial luciferase (lux genes). Hybridization of a lux AB probe from the Kryptophanaron alfredi symbiont to DNAs from 9 strains (8 species) of luminous bacteria showed that none of the strains tested had lux genes highly similar to the symbiont. The most similar were a group consisting of Vibrio harveyi, Vibrio splendidus and Vibrio orientalis. The nucleotide sequence of the luciferase alpha subunit gene luxA) of the Kryptophanaron alfredi symbiont was determined in order to do a more detailed comparison with published luxA sequences from Vibrio harveyi, Vibrio fischeri and Photobacterium leiognathi. The hybridization results, sequence comparisons and the mol% G + C of the Kryptophanaron alfredi symbiont luxA gene suggest that the symbiont may be considered as a new species of luminous Vibrio related to Vibrio harveyi.

  2. Polycyclic aromatic hydrocarbons and trace elements bounded to airborne PM10 in the harbor of Volos, Greece: Implications for the impact of harbor activities

    Science.gov (United States)

    Manoli, E.; Chelioti-Chatzidimitriou, A.; Karageorgou, K.; Kouras, A.; Voutsa, D.; Samara, C.; Kampanos, I.

    2017-10-01

    Harbors are often characterized by high levels of air pollutants that are emitted from ship traffic and other harbor activities. In the present study, the concentrations of Polycyclic Aromatic Hydrocarbons (PAHs) and trace elements (As, Cd, Ni, Pb, Cr, Mn, Zn, and Fe) bounded to the inhalable particulate matter PM10 were studied in the harbor of Volos, central Greece, during a 2-year period (2014-2015). Seasonal and daily variations were investigated. Moreover, total carcinogenic and mutagenic activities of PAHs were calculated. The effect of major wind sectors (sea, city, industrial, harbor) was estimated to assess the potential contribution of ship traffic and harbor activities, such as scrap metal handling operations. Results showed that the harbor sector (calm winds ≤ 0.5 m s-1) was associated with the highest concentrations of PM10. The harbor sector was also associated with relatively increased levels of trace elements (As, Fe, Cr, Mn, Ni), however the effect of this sector was lower than the corresponding effect of the industrial wind sector. The sea sector showed only a slight increase in B[a]Py and Σ12PAHs, whereas the highest increasing effect for PAHs and traffic-related elements, such as Pb and Zn, was evidenced for the city sector.

  3. CC8 MRSA strains harboring SCCmec type IVc are predominant in Colombian hospitals.

    Directory of Open Access Journals (Sweden)

    J Natalia Jiménez

    Full Text Available BACKGROUND: Recent reports highlight the incursion of community-associated MRSA within healthcare settings. However, knowledge of this phenomenon remains limited in Latin America. The aim of this study was to evaluate the molecular epidemiology of MRSA in three tertiary-care hospitals in Medellín, Colombia. METHODS: An observational cross-sectional study was conducted from 2008-2010. MRSA infections were classified as either community-associated (CA-MRSA or healthcare-associated (HA-MRSA, with HA-MRSA further classified as hospital-onset (HAHO-MRSA or community-onset (HACO-MRSA according to standard epidemiological definitions established by the U.S. Centers for Disease Control and Prevention (CDC. Genotypic analysis included SCCmec typing, spa typing, PFGE and MLST. RESULTS: Out of 538 total MRSA isolates, 68 (12.6% were defined as CA-MRSA, 243 (45.2% as HACO-MRSA and 227 (42.2% as HAHO-MRSA. The majority harbored SCCmec type IVc (306, 58.7%, followed by SCCmec type I (174, 33.4%. The prevalence of type IVc among CA-, HACO- and HAHO-MRSA isolates was 92.4%, 65.1% and 43.6%, respectively. From 2008 to 2010, the prevalence of type IVc-bearing strains increased significantly, from 50.0% to 68.2% (p = 0.004. Strains harboring SCCmec IVc were mainly associated with spa types t1610, t008 and t024 (MLST clonal complex 8, while PFGE confirmed that the t008 and t1610 strains were closely related to the USA300-0114 CA-MRSA clone. Notably, strains belonging to these three spa types exhibited high levels of tetracycline resistance (45.9%. CONCLUSION: CC8 MRSA strains harboring SCCmec type IVc are becoming predominant in Medellín hospitals, displacing previously reported CC5 HA-MRSA clones. Based on shared characteristics including SCCmec IVc, absence of the ACME element and tetracycline resistance, the USA300-related isolates in this study are most likely related to USA300-LV, the recently-described 'Latin American variant' of USA300.

  4. Response to crizotinib in a lung adenocarcinoma patient harboring a novel SLC34A2-ROS1 fusion variant

    Science.gov (United States)

    Zhao, Zheng; Song, Zhangjun; Wang, Xuwei; Sun, Haifeng; Yang, Xiaomin; Yuan, Yong; Yu, Pan

    2017-01-01

    ROS1 fusion is a common genetic alteration in non-small-cell lung cancer. Crizotinib, an anaplastic lymphoma kinase inhibitor, shows efficacy in the treatment of lung cancer cases with ROS1 translocation. We report the response to crizotinib of a lung adenocarcinoma patient harboring a novel SLC34A2-ROS1 fusion variant, which was different from the two common SLC34A2-ROS1 fusion types reported in the literature. After crizotinib administration, overall recovery was good in this patient; the primary lesion was successfully treated, the lymph node metastases had disappeared, and the metabolism was normal. PMID:28860822

  5. Influenza A(H10N7) Virus in Dead Harbor Seals, Denmark

    DEFF Research Database (Denmark)

    Krog, Jesper Schak; Hansen, Mette Sif; Holm, Elisabeth

    2015-01-01

    Since April 2014, an outbreak of influenza in harbor seals has been ongoing in northern Europe. In Denmark during June-August, 152 harbor seals on the island of Anholt were found dead from severe pneumonia. We detected influenza A(H10N7) virus in 2 of 4 seals examined.......Since April 2014, an outbreak of influenza in harbor seals has been ongoing in northern Europe. In Denmark during June-August, 152 harbor seals on the island of Anholt were found dead from severe pneumonia. We detected influenza A(H10N7) virus in 2 of 4 seals examined....

  6. Ecological evaluation of proposed dredged material from Richmond Harbor

    Energy Technology Data Exchange (ETDEWEB)

    Pinza, M R; Ward, J A; Mayhew, H L; Word, J Q; Niyogi, D K; Kohn, N P [Battelle/Marine Sciences Lab., Sequim, WA (United States)

    1992-10-01

    During the summer of 1991, Battelle/Marine Sciences Laboratory (MSL) was contracted to conduct sampling and testing of sediments proposed for dredging of Richmond Harbor, California. The MSL collected sediment cores to a depth of [minus]40 ft MLLW ([minus]38 ft + 2 ft overdepth) from 28 (12-in. core) and 30 (4-in. core) stations. The sediment cores were allocated to six composite samples referred to as sediment treatments, which were then subjected to physical, chemical, toxicological, and bioaccumulation testing. Physical and chemical parameters included grain size, total organic carbon (TOC), total volatile solids (TVS), oil and grease, total petroleum hydrocarbons (TPH), polynuclear aromatic hydrocarbons (PAH), chlorinated pesticides, polychlorinated biphenyis (PCBs), priority pollutant metals, and butyltins. The results from the test treatments were compared to results from five reference treatments representative of potential in-bay and offshore disposal sites.

  7. Ecological evaluation of proposed dredged material from Richmond Harbor

    International Nuclear Information System (INIS)

    Pinza, M.R.; Ward, J.A.; Mayhew, H.L.; Word, J.Q.; Niyogi, D.K.; Kohn, N.P.

    1992-10-01

    During the summer of 1991, Battelle/Marine Sciences Laboratory (MSL) was contracted to conduct sampling and testing of sediments proposed for dredging of Richmond Harbor, California. The MSL collected sediment cores to a depth of -40 ft MLLW (-38 ft + 2 ft overdepth) from 28 (12-in. core) and 30 (4-in. core) stations. The sediment cores were allocated to six composite samples referred to as sediment treatments, which were then subjected to physical, chemical, toxicological, and bioaccumulation testing. Physical and chemical parameters included grain size, total organic carbon (TOC), total volatile solids (TVS), oil and grease, total petroleum hydrocarbons (TPH), polynuclear aromatic hydrocarbons (PAH), chlorinated pesticides, polychlorinated biphenyis (PCBs), priority pollutant metals, and butyltins. The results from the test treatments were compared to results from five reference treatments representative of potential in-bay and offshore disposal sites

  8. West Nile Flavivirus Polioencephalomyelitis in a harbor seal (Phoca vitulina).

    Science.gov (United States)

    Del Piero, F; Stremme, D W; Habecker, P L; Cantile, C

    2006-01-01

    A 12-year-old male harbor seal presented with progressive signs of neurologic dysfunction including head tremors, muzzle twitching, clonic spasms, and weakness. Lesions included polioencephalomyelitis with glial nodules, spheroids, neuronophagia, ring hemorrhages, and a few neutrophils. Neurons, fibers, and glial nodules were multifocally colonized with intracytoplasmic West Nile flavivirus antigens that were demonstrated using indirect immunohistochemical analysis. Flavivirus on cultured cells also was isolated and was identified by use of monoclonal antibodies and reverse transcriptase-polymerase chain reaction analysis. Clinical signs of disease and lesion morphology and distribution were similar to those of equine West Nile virus infection. Similar to horses, alpacas, humans, dogs, and reptiles, seals can be dead-end hosts of West Nile virus.

  9. Structural damage monitoring of harbor caissons with interlocking condition

    Energy Technology Data Exchange (ETDEWEB)

    Huynh, Thanh Canh; Lee, So Young; Nauyen, Khac Duy; Kim, Jeong Tae [Pukyong National Univ., Busan (Korea, Republic of)

    2012-12-15

    The objective of this study is to monitor the health status of harbor caissons which have potential foundation damage. To obtain the objective, the following approaches are performed. Firstly, a structural damage monitoring(SDM) method is designed for interlocked multiple caisson structures. The SDM method utilizes the change in modal strain energy to monitor the foundation damage in a target caisson unit. Secondly, a finite element model of a caisson system which consists of three caisson units is established to verify the feasibility of the proposed method. In the finite element simulation, the caisson units are constrained each other by shear key connections. The health status of the caisson system against various levels of foundation damage is monitored by measuring relative modal displacements between the adjacent caissons.

  10. Structural damage monitoring of harbor caissons with interlocking condition

    International Nuclear Information System (INIS)

    Huynh, Thanh Canh; Lee, So Young; Nauyen, Khac Duy; Kim, Jeong Tae

    2012-01-01

    The objective of this study is to monitor the health status of harbor caissons which have potential foundation damage. To obtain the objective, the following approaches are performed. Firstly, a structural damage monitoring(SDM) method is designed for interlocked multiple caisson structures. The SDM method utilizes the change in modal strain energy to monitor the foundation damage in a target caisson unit. Secondly, a finite element model of a caisson system which consists of three caisson units is established to verify the feasibility of the proposed method. In the finite element simulation, the caisson units are constrained each other by shear key connections. The health status of the caisson system against various levels of foundation damage is monitored by measuring relative modal displacements between the adjacent caissons

  11. Geographic, seasonal, and diurnal surface behavior of harbor porpoises

    DEFF Research Database (Denmark)

    Teilmann, Jonas; Christiansen, C.T.; Kjellerup, Sanne

    2013-01-01

    are essential information on the status and management of the species. Thirty-five free-ranging harbor porpoises (Phocoena phocoena) were tracked in the region between the Baltic and the North Sea for 25-349 d using Argos satellite transmitters. No differences were found in surface behavior between geographical...... areas or the size of the animals. Slight differences were found between the two sexes and time of day. Surface time peaked in April, where 6% was spent with the transmitter above surface and 61.5% between 0 and 2 m depth, while the minimum values occurred in February (3.4% and 42.5%, respectively......). The analyses reveal that individual variation among porpoises is the most important factor in explaining variation in surface rates. However, the large number of animals documented in the present study covering a wide range of age and sex groups justifies the use of the seasonal average surface times...

  12. Anthropogenic inputs of dissolved organic matter in New York Harbor

    Science.gov (United States)

    Gardner, G. B.; Chen, R. F.; Olavasen, J.; Peri, F.

    2016-02-01

    The Hudson River flows into the Atlantic Ocean through a highly urbanized region which includes New York City to the east and Newark, New Jersey to the west. As a result, the export of Dissolved Organic Carbon (DOC) from the Hudson to the Atlantic Ocean includes a significant anthropogenic component. A series of high resolution studies of the DOC dynamics of this system were conducted between 2003 and 2010. These included both the Hudson and adjacent large waterways (East River, Newark Bay, Kill Van Kull and Arthur Kill) using coastal research vessels and smaller tributaries (Hackensack, Pasaic and Raritan rivers) using a 25' boat. In both cases measurements were made using towed instrument packages which could be cycled from near surface to near bottom depths with horizontal resolution of approximately 20 to 200 meters depending on depth and deployment strategy. Sensors on the instrument packages included a CTD to provide depth and salinity information and a chromophoric dissolved organic matter(CDOM) fluorometer to measure the fluorescent fraction of the DOC. Discrete samples allowed calibration of the fluorometer and the CDOM data to be related to DOC. The combined data set from these cruises identified multiple scales of source and transport processes for DOC within the Hudson River/New York Harbor region. The Hudson carries a substantial amount of natural DOC from its 230 km inland stretch. Additional sources exist in fringing salt marshes adjacent to the Hackensack and Raritan rivers. However the lower Hudson/New Harbor region receives a large input of DOC from multiple publically owned treatment works (POTW) discharges. The high resolution surveys allowed us to elucidate the distribution of these sources and the manner in which they are rapidly mixed to create the total export. We estimate that anthropogenic sources account for up to 2.5 times the DOC flux contributed by natural processes.

  13. Preliminary Feasibility Report (Stage 2), Review of Reports on Lorain Harbor, Ohio. Volume 2. Appendices. Revision

    Science.gov (United States)

    1981-05-01

    casts for iron ore within the GL/SLS region. A recent downturn in the economic health of the domestic steel industry has probably deferred any major...emergents: Swamp rose mallow Hibiscus palustris Nettle Urtica sp. Nightshade Solanum dulcamara Hedge bindweed Convolvulus sepium Peppermint Mentha arvensis

  14. The hearing threshold of a harbor porpoise (Phocoena phocoena) for impulsive sounds (L)

    NARCIS (Netherlands)

    Kastelein, R.A.; Gransier, R.; Hoek, L.; Jong, C.A.F. de

    2012-01-01

    The distance at which harbor porpoises can hear underwater detonation sounds is unknown, but depends, among other factors, on the hearing threshold of the species for impulsive sounds. Therefore, the underwater hearing threshold of a young harbor porpoise for an impulsive sound, designed to mimic a

  15. 76 FR 38302 - Safety Zone; Cape Charles Fireworks, Cape Charles Harbor, Cape Charles, VA

    Science.gov (United States)

    2011-06-30

    ... the Town of Cape Charles will sponsor a fireworks display on the shoreline of the navigable waters of...-AA00 Safety Zone; Cape Charles Fireworks, Cape Charles Harbor, Cape Charles, VA AGENCY: Coast Guard... navigable waters of Cape Charles City Harbor in Cape Charles, VA in support of the Fourth of July Fireworks...

  16. 77 FR 29929 - Safety Zone; Town of Cape Charles Fireworks, Cape Charles Harbor, Cape Charles, VA

    Science.gov (United States)

    2012-05-21

    ... section of this notice. Basis and Purpose On July 4, 2012 the Town of Cape Charles will sponsor a...-AA00 Safety Zone; Town of Cape Charles Fireworks, Cape Charles Harbor, Cape Charles, VA AGENCY: Coast... temporary safety zone on the waters of Cape Charles City Harbor in Cape Charles, VA in support of the Fourth...

  17. 76 FR 71598 - Pearl Harbor National Wildlife Refuge, Honolulu County, HI; Final Comprehensive Conservation Plan...

    Science.gov (United States)

    2011-11-18

    ...] Pearl Harbor National Wildlife Refuge, Honolulu County, HI; Final Comprehensive Conservation Plan and.../docs/HI-PI/docsjcpearl.htm . Email: [email protected] . Include ``Pearl Harbor final CCP'' in...`iwa, HI 96712. FOR FURTHER INFORMATION CONTACT: David Ellis, Project Leader, (808) 637-6330...

  18. 33 CFR 110.129a - Apra Harbor, Guam. (Datum: WGS 84)

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Apra Harbor, Guam. (Datum: WGS 84) 110.129a Section 110.129a Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY ANCHORAGES ANCHORAGE REGULATIONS Special Anchorage Areas § 110.129a Apra Harbor, Guam. (Datum: WGS 84) (a...

  19. 33 CFR 80.1490 - Apra Harbor, U.S. Territory of Guam.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Apra Harbor, U.S. Territory of Guam. 80.1490 Section 80.1490 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY INTERNATIONAL NAVIGATION RULES COLREGS DEMARCATION LINES Pacific Islands § 80.1490 Apra Harbor, U...

  20. 76 FR 34865 - Safety Zone; Rochester Harbor Festival, Genesee River, Rochester, NY

    Science.gov (United States)

    2011-06-15

    ...-AA00 Safety Zone; Rochester Harbor Festival, Genesee River, Rochester, NY AGENCY: Coast Guard, DHS. ACTION: Temporary final rule. SUMMARY: The Coast Guard is establishing a temporary safety zone on the Genesee River, Rochester, NY for the Rochester Harbor Festival fireworks. This zone is intended to...

  1. 33 CFR 110.31 - Hull Bay and Allerton Harbor at Hull, Mass.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Hull Bay and Allerton Harbor at Hull, Mass. 110.31 Section 110.31 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY ANCHORAGES ANCHORAGE REGULATIONS Special Anchorage Areas § 110.31 Hull Bay and Allerton Harbor at...

  2. 76 FR 81904 - Solicitation of New Safe Harbors and Special Fraud Alerts

    Science.gov (United States)

    2011-12-29

    ... Medicaid Patient and Program Protection Act of 1987, Public Law 100-93 Sec. 14, the Act, Sec. 1128B(b), 42...-called ``safe harbor'' provisions, specifying various payment and business practices that, although... basis for administrative sanctions. OIG safe harbor provisions have been developed ``to limit the reach...

  3. 33 CFR 117.753 - Ship Channel, Great Egg Harbor Bay.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Ship Channel, Great Egg Harbor Bay. 117.753 Section 117.753 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY BRIDGES DRAWBRIDGE OPERATION REGULATIONS Specific Requirements New Jersey § 117.753 Ship Channel, Great Egg Harbor Bay. The draw of the S52 (Ship...

  4. 33 CFR 117.458 - Inner Harbor Navigation Canal, New Orleans.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Inner Harbor Navigation Canal, New Orleans. 117.458 Section 117.458 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF... Harbor Navigation Canal, New Orleans. (a) The draws of the SR 46 (St. Claude Avenue) bridge, mile 0.5...

  5. 77 FR 46285 - Drawbridge Operation Regulation; Inner Harbor Navigational Canal, New Orleans, LA

    Science.gov (United States)

    2012-08-03

    ... Operation Regulation; Inner Harbor Navigational Canal, New Orleans, LA AGENCY: Coast Guard, DHS. ACTION... Navigational Canal, mile 3.1, at New Orleans, LA. The deviation is necessary to replace the wire rope lifting... of the US 90 (Danzinger) Bridge across the Inner Harbor Navigational Canal, mile 3.1, at New Orleans...

  6. 75 FR 4693 - Drawbridge Operation Regulation; Inner Harbor Navigational Canal, New Orleans, LA

    Science.gov (United States)

    2010-01-29

    ... Operation Regulation; Inner Harbor Navigational Canal, New Orleans, LA AGENCY: Coast Guard, DHS. ACTION... C. Simon) Bascule Bridge across the Inner Harbor Navigational Canal, mile 4.6, at New Orleans, LA... Ochsner Ironman 70.3 New Orleans event. This deviation allows the bridge to remain closed during the event...

  7. 76 FR 37005 - Safety Zone; Fan Pier Yacht Club Fireworks, Boston Harbor, Boston, MA

    Science.gov (United States)

    2011-06-24

    ...-AA00 Safety Zone; Fan Pier Yacht Club Fireworks, Boston Harbor, Boston, MA AGENCY: Coast Guard, DHS... Sector Boston Captain of the Port (COTP) Zone for the Fan Pier Yacht Club Fireworks display. This safety... Safety Zone; Fan Pier Yacht Club Fireworks, Boston Harbor, Boston, Massachusetts. (a) General. A...

  8. 33 CFR 110.27 - Lynn Harbor in Broad Sound, Mass.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Lynn Harbor in Broad Sound, Mass. 110.27 Section 110.27 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY ANCHORAGES ANCHORAGE REGULATIONS Special Anchorage Areas § 110.27 Lynn Harbor in Broad Sound, Mass. North of...

  9. Analyzing Approaches to Internet Jurisdiction Based on Model of Harbors and the High Seas

    NARCIS (Netherlands)

    Jiménez, W.G.; Lodder, A.R.

    2015-01-01

    The inherent cross-border nature of the internet has challenged the legal system for over two decades. In this paper we introduce a model in which the internet is approached as if it were the high seas, the harbor of origin, the harbor of destination, or a combination of these. This model is used to

  10. 33 CFR 165.754 - Safety Zone: San Juan Harbor, San Juan, PR.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Safety Zone: San Juan Harbor, San Juan, PR. 165.754 Section 165.754 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND... Zone: San Juan Harbor, San Juan, PR. (a) Regulated area. A moving safety zone is established in the...

  11. Genetic characterization of blaNDM-harboring plasmids in carbapenem-resistant Escherichia coli from Myanmar.

    Directory of Open Access Journals (Sweden)

    Yo Sugawara

    Full Text Available The bacterial enzyme New Delhi metallo-β-lactamase hydrolyzes almost all β-lactam antibiotics, including carbapenems, which are drugs of last resort for severe bacterial infections. The spread of carbapenem-resistant Enterobacteriaceae that carry the New Delhi metallo-β-lactamase gene, blaNDM, poses a serious threat to public health. In this study, we genetically characterized eight carbapenem-resistant Escherichia coli isolates from a tertiary care hospital in Yangon, Myanmar. The eight isolates belonged to five multilocus-sequence types and harbored multiple antimicrobial-resistance genes, resulting in resistance against nearly all of the antimicrobial agents tested, except colistin and fosfomycin. Nine plasmids harboring blaNDM genes were identified from these isolates. Multiple blaNDM genes were found in the distinct Inc-replicon types of the following plasmids: an IncA/C2 plasmid harboring blaNDM-1 (n = 1, IncX3 plasmids harboring blaNDM-4 (n = 2 or blaNDM-7 (n = 1, IncFII plasmids harboring blaNDM-4 (n = 1 or blaNDM-5 (n = 3, and a multireplicon F plasmid harboring blaNDM-5 (n = 1. Comparative analysis highlighted the diversity of the blaNDM-harboring plasmids and their distinct characteristics, which depended on plasmid replicon types. The results indicate circulation of phylogenetically distinct strains of carbapenem-resistant E. coli with various plasmids harboring blaNDM genes in the hospital.

  12. 77 FR 2019 - Security Zone; Passenger Vessel SAFARI EXPLORER Arrival/Departure, Kaunakakai Harbor, Molokai, HI

    Science.gov (United States)

    2012-01-13

    ...-AA87 Security Zone; Passenger Vessel SAFARI EXPLORER Arrival/ Departure, Kaunakakai Harbor, Molokai, HI... vessel SAFARI EXPLORER to its intended berth in the harbor. Entry into the temporary security zone is... operation of the SAFARI EXPLORER into Molokai has been voluntarily suspended by the operating company...

  13. 77 FR 24381 - Security Zone; Passenger Vessel SAFARI EXPLORER Arrival/Departure, Kaunakakai Harbor, Molokai, HI

    Science.gov (United States)

    2012-04-24

    ...-AA87 Security Zone; Passenger Vessel SAFARI EXPLORER Arrival/ Departure, Kaunakakai Harbor, Molokai, HI... channel's entrance during the arrival and departure of the Passenger Vessel Safari Explorer in Kaunakakai... entrance during the arrival and departure of the Passenger Vessel Safari Explorer in Kaunakakai Harbor...

  14. 33 CFR 110.235 - Pacific Ocean (Mamala Bay), Honolulu Harbor, Hawaii (Datum: NAD 83).

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Pacific Ocean (Mamala Bay), Honolulu Harbor, Hawaii (Datum: NAD 83). 110.235 Section 110.235 Navigation and Navigable Waters COAST... Pacific Ocean (Mamala Bay), Honolulu Harbor, Hawaii (Datum: NAD 83). (a) The anchorage grounds—(1...

  15. Environmental space management in the harbor of Amsterdam, Netherlands; Milieuruimtemanagement haven Amsterdam

    Energy Technology Data Exchange (ETDEWEB)

    Klok, L.; Hulskotte, J. [TNO Built Environment and Geosciences, Den Haag (Netherlands); Van Breemen, T. [Haven Amsterdam, Amsterdam (Netherlands)

    2012-02-15

    A new calculation tool will quickly offer the Harbor of Amsterdam insight in the effect of activities in the harbor on the air quality and hence the available environmental space. [Dutch] Een nieuw rekeninstrument geeft Haven Amsterdam snel inzicht in het effect van alle activiteiten in de haven op de luchtkwaliteit en daarmee in de beschikbare milieuruimte.

  16. 75 FR 81556 - Solicitation of New Safe Harbors and Special Fraud Alerts

    Science.gov (United States)

    2010-12-28

    ... statute (section 1128B(b) of the Social Security Act), as well as developing new OIG Special Fraud Alerts... of New Safe Harbors and Special Fraud Alerts AGENCY: Office of Inspector General (OIG), HHS. ACTION... the public on recommendations for developing new or revised safe harbors and Special Fraud Alerts...

  17. 77 FR 76434 - Solicitation of New Safe Harbors and Special Fraud Alerts

    Science.gov (United States)

    2012-12-28

    ... statute (section 1128B(b) of the Social Security Act), as well as developing new OIG Special Fraud Alerts... of New Safe Harbors and Special Fraud Alerts AGENCY: Office of Inspector General (OIG), HHS. ACTION... revised safe harbors and Special Fraud Alerts. Please assist us by referencing the file code OIG-121-N...

  18. 78 FR 78807 - Solicitation of New Safe Harbors and Special Fraud Alerts

    Science.gov (United States)

    2013-12-27

    ... statute (section 1128B(b) of the Social Security Act), as well as developing new OIG Special Fraud Alerts... of New Safe Harbors and Special Fraud Alerts AGENCY: Office of Inspector General (OIG), HHS. ACTION... revised safe harbors and Special Fraud Alerts. Please assist us by referencing the file code OIG-122-N...

  19. 33 CFR 207.600 - Rochester (Charlotte) Harbor, N.Y.; use, administration, and navigation.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Rochester (Charlotte) Harbor, N.Y.; use, administration, and navigation. 207.600 Section 207.600 Navigation and Navigable Waters CORPS OF... (Charlotte) Harbor, N.Y.; use, administration, and navigation. (a)-(b) [Reserved] (c) No vessel shall moor or...

  20. 33 CFR 207.580 - Buffalo Harbor, N.Y.; use, administration, and navigation.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Buffalo Harbor, N.Y.; use, administration, and navigation. 207.580 Section 207.580 Navigation and Navigable Waters CORPS OF ENGINEERS, DEPARTMENT OF THE ARMY, DEPARTMENT OF DEFENSE NAVIGATION REGULATIONS § 207.580 Buffalo Harbor, N.Y.; use...

  1. 78 FR 13479 - Drawbridge Operation Regulation; New Haven Harbor, Quinnipiac and Mill Rivers, CT

    Science.gov (United States)

    2013-02-28

    ...-AA09 Drawbridge Operation Regulation; New Haven Harbor, Quinnipiac and Mill Rivers, CT AGENCY: Coast... regulations that govern the operation of three bridges across the Quinnipiac and Mill Rivers at New Haven...) entitled ``Drawbridge Operation Regulations New Haven Harbor, Quinnipiac and Mill Rivers,'' in the Federal...

  2. Congress Investigates: Pearl Harbor and 9/11 Congressional Hearing Exhibits

    Science.gov (United States)

    Blackerby, Christine

    2011-01-01

    On the morning of December 7, 1941, Japanese bombers staged a surprise attack on U.S. military forces at Pearl Harbor in Hawaii. Sixty years after the attack on Pearl Harbor, the United States was attacked again. On the morning of September 11, 2001, four commercial airplanes hijacked by 19 terrorists killed nearly 3,000 people when they crashed…

  3. Pearl Harbor bombing attack: a contamination legacy revealed in the sedimentary record

    International Nuclear Information System (INIS)

    Ashwood, T.L.; Olsen, C.R.

    1988-01-01

    Sediment cores collected from Middle Loch and West Loch of Pearl Harbor were dated using fallout 137 Cs and excess 210 Pb and analysed for several major elements and trace metals. Results indicate that sediment deposited immediately following the 7 December, 1941, bombing attack on Pearl Harbor contained elevated levels of several trace metals. (author)

  4. Pearl Harbor bombing attack: a contamination legacy revealed in the sedimentary record

    Energy Technology Data Exchange (ETDEWEB)

    Ashwood, T.L.; Olsen, C.R.

    1988-02-01

    Sediment cores collected from Middle Loch and West Loch of Pearl Harbor were dated using fallout /sup 137/Cs and excess /sup 210/Pb and analysed for several major elements and trace metals. Results indicate that sediment deposited immediately following the 7 December, 1941, bombing attack on Pearl Harbor contained elevated levels of several trace metals.

  5. 77 FR 38490 - Safety Zone; Mentor Harbor Yachting Club Fireworks, Lake Erie, Mentor, OH

    Science.gov (United States)

    2012-06-28

    ...-AA00 Safety Zone; Mentor Harbor Yachting Club Fireworks, Lake Erie, Mentor, OH AGENCY: Coast Guard, DHS... Erie, Mentor, OH. This safety zone is intended to restrict vessels from a portion of Lake Erie during the Mentor Harbor Yachting Club fireworks display. This temporary safety zone is necessary to protect...

  6. Are federal sustained yield units equitable? A case study of the Grays Harbor unit.

    Science.gov (United States)

    Con H Schallau; Wilbur R. Maki

    1986-01-01

    The Grays Harbor Federal Sustained Yield Unit (U.S. Department of Agriculture, Forest Service) was established in 1949 to enhance the economic stability of the forest products industry and dependent communities in Grays Harbor County, Washington. Provisions of the unit's charter require that all logs harvested from the Quinault Ranger District of the Olympic...

  7. 78 FR 19632 - Special Local Regulations; St. Thomas Carnival Watersport Activities, Charlotte Amalie Harbor; St...

    Science.gov (United States)

    2013-04-02

    ...-AA08 Special Local Regulations; St. Thomas Carnival Watersport Activities, Charlotte Amalie Harbor; St... proposes to establish a special local regulation on the waters of Charlotte Amalie Harbor in St Thomas, USVI during the St. Thomas Carnival Watersport Activities, a high speed boat race. The event is...

  8. 33 CFR 110.214 - Los Angeles and Long Beach harbors, California.

    Science.gov (United States)

    2010-07-01

    ... Angeles Harbor). A circular area with a radius of 400 yards (approximately 366 meters), centered in... 400 Transportation Corridor. (C) Outer Harbor: The western boundary of Commercial Anchorage B. (2... Thence along a line described as an arc, radius of 460 meters (approximately 1509 feet) centered on 33...

  9. 77 FR 60109 - Takes of Marine Mammals Incidental to Specified Activities; Harbor Activities Related to the...

    Science.gov (United States)

    2012-10-02

    ...; and Northern elephant seals by Level B harassment only. To date, we have issued nine, 1-year... numbers of marine mammals, incidental to conducting Delta Mariner harbor operations for one year. We... California sea lions (Zalophus californianus), Pacific harbor seals (Phoca vitulina), and Northern elephant...

  10. 78 FR 28492 - Special Local Regulation; Low Country Splash, Wando River, Cooper River, and Charleston Harbor...

    Science.gov (United States)

    2013-05-15

    ...-AA08 Special Local Regulation; Low Country Splash, Wando River, Cooper River, and Charleston Harbor... establishing a special local regulation on the waters of the Wando River, Cooper River, and Charleston Harbor... rulemaking (NPRM) entitled, ``Special Local Regulation; Low Country Splash, Wando River, Cooper River, and...

  11. 33 CFR 165.708 - Safety/Security Zone; Charleston Harbor and Cooper River, Charleston, SC.

    Science.gov (United States)

    2010-07-01

    ... Harbor and Cooper River, Charleston, SC. 165.708 Section 165.708 Navigation and Navigable Waters COAST... Guard District § 165.708 Safety/Security Zone; Charleston Harbor and Cooper River, Charleston, SC. (a... Cooper River. All coordinates referenced use datum: NAD 1983. (2) All waters within 100 yards of the...

  12. 33 CFR 110.250 - St. Thomas Harbor, Charlotte Amalie, V.I.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false St. Thomas Harbor, Charlotte... SECURITY ANCHORAGES ANCHORAGE REGULATIONS Anchorage Grounds § 110.250 St. Thomas Harbor, Charlotte Amalie.... Thomas, V.I. of the United States and approaches thereto, including all waters under its jurisdiction, as...

  13. Isolation of Shiga toxin-producing Escherichia coli harboring variant Shiga toxin genes from seafood

    Directory of Open Access Journals (Sweden)

    Sreepriya Prakasan

    2018-03-01

    Full Text Available Background and Aim: Shiga toxin-producing Escherichia coli (STEC are important pathogens of global significance. STEC are responsible for numerous food-borne outbreaks worldwide and their presence in food is a potential health hazard. The objective of the present study was to determine the incidence of STEC in fresh seafood in Mumbai, India, and to characterize STEC with respect to their virulence determinants. Materials and Methods: A total of 368 E. coli were isolated from 39 fresh seafood samples (18 finfish and 21 shellfish using culture-based methods. The isolates were screened by polymerase chain reaction (PCR for the genes commonly associated with STEC. The variant Shiga toxin genes were confirmed by Southern blotting and hybridization followed by DNA sequencing. Results: One or more Shiga toxins genes were detected in 61 isolates. Of 39 samples analyzed, 10 (25.64% samples harbored STEC. Other virulence genes, namely, eaeA (coding for an intimin and hlyA (hemolysin A were detected in 43 and 15 seafood isolates, respectively. The variant stx1 genes from 6 isolates were sequenced, five of which were found to be stx1d variants, while one sequence varied considerably from known stx1 sequences. Southern hybridization and DNA sequence analysis suggested putative Shiga toxin variant genes (stx2 in at least 3 other isolates. Conclusion: The results of this study showed the occurrence of STEC in seafood harboring one or more Shiga toxin genes. The detection of STEC by PCR may be hampered due to the presence of variant genes such as the stx1d in STEC. This is the first report of stx1d gene in STEC isolated from Indian seafood.

  14. Ecological evaluation of proposed discharge of dredged material from Oakland Harbor into ocean waters (Phase 3 B of -42-foot project)

    Energy Technology Data Exchange (ETDEWEB)

    Kohn, N.P.; Ward, J.A.; Mayhew, H.L.; Word, J.Q.; Barrows, E.S.; Goodwin, S.M.; Lefkovitz, L.F. (Battelle/Marine Sciences Lab., Sequim, WA (United States))

    1992-06-01

    The Water Resources Development Act of 1986 (Public Law 99-662) authorized the US Army Corps of Engineers (USACE) San Francisco District, to deepen and widen the navigational channels of the Oakland Inner and Outer Harbors to accommodate deeper-draft vessels. The USACE is considering several disposal options for the dredged material removed during these channel improvements including open-water disposal. Dredged material proposed for open-water disposal must be evaluated to determine the potential impacts of the disposal activity on the water column and disposal site environments. The USACE requested that Battelle/Marine Sciences Laboratory (MSL) conduct studies to evaluate open-water disposal options for Oakland Harbor sediments. This request developed into the Oakland Harbor Phase III Program. This is Volume 2 of a two-volume report that presents information gathered to determine the suitability of ocean disposal of sediments dredged from Oakland Harbor. This volume contains the Appendixes (A through N), which provide details of the data analyses and full presentation of the data and results.

  15. Ecological evaluation of proposed discharge of dredged material from Oakland Harbor into ocean waters (Phase 3 B of -42-foot project). Volume 2, Appendixes

    Energy Technology Data Exchange (ETDEWEB)

    Kohn, N.P.; Ward, J.A.; Mayhew, H.L.; Word, J.Q.; Barrows, E.S.; Goodwin, S.M.; Lefkovitz, L.F. [Battelle/Marine Sciences Lab., Sequim, WA (United States)

    1992-06-01

    The Water Resources Development Act of 1986 (Public Law 99-662) authorized the US Army Corps of Engineers (USACE) San Francisco District, to deepen and widen the navigational channels of the Oakland Inner and Outer Harbors to accommodate deeper-draft vessels. The USACE is considering several disposal options for the dredged material removed during these channel improvements including open-water disposal. Dredged material proposed for open-water disposal must be evaluated to determine the potential impacts of the disposal activity on the water column and disposal site environments. The USACE requested that Battelle/Marine Sciences Laboratory (MSL) conduct studies to evaluate open-water disposal options for Oakland Harbor sediments. This request developed into the Oakland Harbor Phase III Program. This is Volume 2 of a two-volume report that presents information gathered to determine the suitability of ocean disposal of sediments dredged from Oakland Harbor. This volume contains the Appendixes (A through N), which provide details of the data analyses and full presentation of the data and results.

  16. Abstracts of papers presented at the LVIII Cold Spring Harbor Symposium on quantitative Biology: DNA and chromosomes

    Energy Technology Data Exchange (ETDEWEB)

    1993-12-31

    This volume contains the abstracts of oral and poster presentations made at the LVIII Cold Spring Harbor Symposium on Quantitative Biology entitles DNA & Chromosomes. The meeting was held June 2--June 9, 1993 at Cold Spring Harbor, New York.

  17. Strategies for Overcoming Resistance in Tumours Harboring BRAF Mutations

    Directory of Open Access Journals (Sweden)

    Nourah Mohammad Obaid

    2017-03-01

    Full Text Available The development of resistance to previously effective treatments has been a challenge for health care providers and a fear for patients undergoing cancer therapy. This is an unfortunately frequent occurrence for patients undergoing targeted therapy for tumours harboring the activating V600E mutation of the BRAF gene. Since the initial identification of the BRAF mutation in 2002, a series of small molecular inhibitors that target the BRAFV600E have been developed, but intrinsic and acquired resistance to these drugs has presented an ongoing challenge. More recently, improvements in therapy have been achieved by combining the use of BRAF inhibitors with other drugs, such as inhibitors of the downstream effector mitogen activated protein kinase (MAPK/extracellular-signal regulated kinase (ERK kinase (MEK. Despite improved success in response rates and in delaying resistance using combination therapy, ultimately, the acquisition of resistance remains a concern. Recent research articles have shed light on some of the underlying mechanisms of this resistance and have proposed numerous strategies that might be employed to overcome or avoid resistance to targeted therapies. This review will explore some of the resistance mechanisms, compare what is known in melanoma cancer to colorectal cancer, and discuss strategies under development to manage the development of resistance.

  18. Synanthropic Cockroaches (Blattidae: Periplaneta spp.) Harbor Pathogenic Leptospira in Colombia.

    Science.gov (United States)

    Gonzalez-Astudillo, Viviana; Bustamante-Rengifo, Javier A; Bonilla, Álvaro; Lehmicke, Anna Joy J; Castillo, Andrés; Astudillo-Hernández, Miryam

    2016-01-01

    Leptospirosis cases in Colombia are typically linked to peridomestic rodents; however, empirical data suggest that Leptospira-infected patients with no apparent exposure to these reservoirs are common. Cockroaches (Periplaneta spp.) have equal or greater interaction with humans than rodents, yet their potential role as carriers of Leptospira has not been assessed. We determined if pathogenic Leptospira is harbored by Periplaneta spp. in Cali (Colombia) and the variables influencing this relationship. Fifty-nine cockroaches were captured from seven sites and DNA was extracted from the body surface and digestive tract for a multiplex polymerase chain reaction, targeting genes secY and flaB. Logistic regression models and proportion tests showed a higher likelihood for Leptospira to be isolated from body surfaces (P > 0.001) and from individuals inside houses (six times more likely). These findings are the first to demonstrate an association between Periplaneta spp. and Leptospira, suggesting the need to investigate the potential for cockroaches to serve as reservoirs or transport hosts for Leptospira. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  19. Evaluation of older bay mud sediment from Richmond Harbor, California

    Energy Technology Data Exchange (ETDEWEB)

    Pinza, M.R.; Mayhew, H.L.; Word, J.Q.

    1996-09-01

    The older, bay mud (OBM) unit predates modem man and could act as a barrier to the downward transport of contaminants from the younger bay mud (YBM) because of its hard-packed consistency. However, its chemical and biological nature have not been well characterized. Battelle/Marine Sciences Laboratory (MSL) conducted three independent studies of OBM sediment in January 1993, January 1994, and October 1994. These studies evaluated potential chemical contamination and biological effects of OBM that could occur as a result of dredging and disposal activities. These evaluations were performed by conducting chemical analysis, solid-phase toxicity tests, suspended- particulate-phase (SPP) toxicity tests, and bioaccumulation tests on the OBM sediment. If the sediment chemistry and toxicity results showed no or minimal contamination and toxicological responses, then either the OBM could be left exposed in Richmond Harbor after dredging the YBM without leaving a source of contamination, or if the project depths necessitate, the OBM would be acceptable for disposal at an appropriate disposal site.

  20. The yeast genome may harbor hypoxia response elements (HRE).

    Science.gov (United States)

    Ferreira, Túlio César; Hertzberg, Libi; Gassmann, Max; Campos, Elida Geralda

    2007-01-01

    The hypoxia-inducible factor-1 (HIF-1) is a heterodimeric transcription factor activated when cells are submitted to hypoxia. The heterodimer is composed of two subunits, HIF-1alpha and the constitutively expressed HIF-1beta. During normoxia, HIF-1alpha is degraded by the 26S proteasome, but hypoxia causes HIF-1alpha to be stabilized, enter the nucleus and bind to HIF-1beta, thus forming the active complex. The complex then binds to the regulatory sequences of various genes involved in physiological and pathological processes. The specific regulatory sequence recognized by HIF-1 is the hypoxia response element (HRE) that has the consensus sequence 5'BRCGTGVBBB3'. Although the basic transcriptional regulation machinery is conserved between yeast and mammals, Saccharomyces cerevisiae does not express HIF-1 subunits. However, we hypothesized that baker's yeast has a protein analogous to HIF-1 which participates in the response to changes in oxygen levels by binding to HRE sequences. In this study we screened the yeast genome for HREs using probabilistic motif search tools. We described 24 yeast genes containing motifs with high probability of being HREs (p-value<0.1) and classified them according to biological function. Our results show that S. cerevisiae may harbor HREs and indicate that a transcription factor analogous to HIF-1 may exist in this organism.

  1. Foraminiferal proxies for pollution monitoring in moderately polluted harbors

    International Nuclear Information System (INIS)

    Armynot du Chatelet, E.; Debenay, J.-P.; Soulard, R.

    2004-01-01

    Foraminiferal density and species richness that decrease with an increase in heavy metal and PAH concentration may be used as pollution indicators. - Benthic foraminifera are increasingly used as environmental bio-indicators, especially in polluted environments where their sensitivity to pollutants may be expressed by a modification of the assemblages. Eighteen sediment samples were collected in September 2000 in five harbors located in moderately polluted estuaries on the coast of Vendee (France) for the study of foraminiferal assemblages. Ten heavy metals and 13 PAH have been analyzed from the sediments. The marine to continental estuarine gradient has a prevalent influence on the foraminiferal distribution. However, the results show that foraminiferal density and species richness of the assemblages decrease with an increase in heavy metal and PAH concentration, and therefore may be used as pollution indicators. Moreover, the more polluted areas are dominated by the tolerant pioneer species Haynesina germanica that may be used as bio-indicator of pollution, mainly in the uppermost areas

  2. In vitro gene imaging by luciferase to detect the expression and effect of human tumor necrosis factor related apoptosis-inducing ligand in lung cancer A549 cells

    International Nuclear Information System (INIS)

    Zhao Na; Cui Jianling; Guo Zhiyuan; Guo Zhiping; Sun Yingcai; Liu Jicun

    2009-01-01

    Objective: To detect the expression and effect of human tumor necrosis factor related apoptosis-inducing ligand(hTRAIL) in vitro by using a novel double expressing adenoviral vector encoding hTRAIL and firefly luciferase (luc) gene (Ad-hTRAIL-luc), in which luc was used as reporter gene. Methods: A549 cells were transduced with the adenoviral vector encoding enhanced green fluorescent protein (EGFP) gene (Ad-EGFP) at variable multiplicity of infection (MOI). Adenoviral transduction efficiency was determined 48 h later. A549 cells were transduced with Ad-hTRAIL-luc at variable MOI, and the following tests were performed 48h later, respectively: the expressive ratio of hTRAIL and the apoptotic ratio of A549 cells were measured by flow cytometer; counts per minute (cpm) of luminescence were measured by scintillation counters. A549 cells were transduced with Ad-luc at variable MOI, and cpm of luminescence was measured by scintillation counters 48 h later. After A549 cells were transduced with Ad-hTRAIL-luc, the expressive ratio of hTRAIL, the apoptotic ratio of A549 cells and cpm of luminescence were analyzed by one-way ANOVA. The positive ratio of EGFP and cpm of luminescence (Ad-luc) were analyzed by nonparametric ANOVA. Results: After A549 cells were transfected with Ad-hTRAIL-luc, the expressive ratio of hTRAIL on the cell membrane of the groups were (2.37±0.04)%, (3.16±0.03)%, (3.64± 0.03)%, (3.96±0.02)%, (4.24±0.02)%, (4.34±0.02)% respectively, which showed significant difference between each other (P<0.01); and the apoptotic ratio of A549 cells were (1.52±0.04)%, (2.93±0.02)%, (3.39±0.02)%, (3.64±0.02)%, (3.86±0.02)%, (4.08±0.02)%, (4.20± 0.02)%, respectively, and it showed significant difference between each other (P<0.01); cpm of luminescence were 465 561 ± 26 801, 1 038 576 ± 29 417, 937 655 ± 23 197, 786 432 ± 20 028, 524 288 ± 16 338, 401 566 ± 15 961, respectively, and it also showed significant difference between each other (P<0

  3. 77 FR 42076 - Notice of Opportunity for Public Comment on Surplus Property Release at Hancock County-Bar Harbor...

    Science.gov (United States)

    2012-07-17

    ... Comment on Surplus Property Release at Hancock County-Bar Harbor Airport, Trenton, ME AGENCY: Federal... located at Hancock County-Bar Harbor Airport, Trenton, Maine. DATES: Comments must be received on or... INFORMATION: The Federal Aviation Administration is reviewing a request by Hancock County-Bar Harbor Airport...

  4. 33 CFR 165.T14-204 - Safety Zone; fixed mooring balls, south of Barbers Pt Harbor Channel, Oahu, Hawaii.

    Science.gov (United States)

    2010-07-01

    ..., south of Barbers Pt Harbor Channel, Oahu, Hawaii. 165.T14-204 Section 165.T14-204 Navigation and... Pt Harbor Channel, Oahu, Hawaii. (a) Location. The following area is a safety zone: All waters... position is approximately 2,500 yards south of Barbers Point Harbor channel buoy #2, Oahu, Hawaii. This...

  5. 77 FR 67563 - Regulated Navigation Area-New Haven Harbor, Quinnipiac River, Mill River, New Haven, CT; Pearl...

    Science.gov (United States)

    2012-11-13

    ... 1625-AA11 Regulated Navigation Area--New Haven Harbor, Quinnipiac River, Mill River, New Haven, CT... Haven Harbor, Quinnipiac River and Mill River. The current RNA pertains only to the operation of tugs...) entitled Regulated Navigation Area--New Haven Harbor, Quinnipiac River, Mill River, New Haven, CT; Pearl...

  6. A lifelong journey of moving beyond wartime trauma for survivors from Hiroshima and Pearl Harbor.

    Science.gov (United States)

    Liehr, Patricia; Nishimura, Chie; Ito, Mio; Wands, Lisa Marie; Takahashi, Ryutaro

    2011-01-01

    This study examines 51 stories of health, shared by people who survived the wartime trauma of Hiroshima and Pearl Harbor, seeking to identify turning points that moved participants along over their lifetime. The central turning point for Hiroshima survivors was "becoming Hibabusha (A-bomb survivor)" and for Pearl Harbor survivors was "honoring the memory and setting it aside." Wartime trauma was permanently integrated into survivors' histories, surfacing steadily over decades for Hiroshima survivors and intermittently over decades for Pearl Harbor survivors. Regardless of experience or nationality, participants moved through wartime trauma by connecting with others, pursuing personal and global peace.

  7. Temporal and spatial variation in harbor seal (Phoca vitulina L.) roar calls from southern Scandinavia

    DEFF Research Database (Denmark)

    Sabinsky, Puk Faxe; Larsen, Ole Næsbye; Wahlberg, Magnus

    2017-01-01

    Male harbor seals gather around breeding sites for competitive mating displays. Here, they produce underwater vocalizations possibly to attract females and/or scare off other males. These calls offer prospects for passive acoustic monitoring. Acoustic monitoring requires a good understanding...... of natural variation in calling behavior both temporally and among geographically separate sites. Such variation in call structure and calling patterns were studied in harbor seal vocalizations recorded at three locations in Danish and Swedish waters. There was a strong seasonality in the calls from end...... biological differences when comparing harbor seal roars among recording sites and between years....

  8. Bioluminescence of beetle luciferases with 6'-amino-D-luciferin analogues reveals excited keto-oxyluciferin as the emitter and phenolate/luciferin binding site interactions modulate bioluminescence colors.

    Science.gov (United States)

    Viviani, Vadim R; Neves, Deimison Rodrigues; Amaral, Danilo Trabuco; Prado, Rogilene A; Matsuhashi, Takuto; Hirano, Takashi

    2014-08-19

    Beetle luciferases produce different bioluminescence colors from green to red using the same d-luciferin substrate. Despite many studies of the mechanisms and structural determinants of bioluminescence colors with firefly luciferases, the identity of the emitters and the specific active site interactions responsible for bioluminescence color modulation remain elusive. To address these questions, we analyzed the bioluminescence spectra with 6'-amino-D-luciferin (aminoluciferin) and its 5,5-dimethyl analogue using a set of recombinant beetle luciferases that naturally elicit different colors and different pH sensitivities (pH-sensitive, Amydetes vivianii λmax=538 nm, Macrolampis sp2 λmax=564 nm; pH-insensitive, Phrixotrix hirtus λmax=623 nm, Phrixotrix vivianii λmax=546 nm, and Pyrearinus termitilluminans λmax=534 nm), a luciferase-like enzyme (Tenebrionidae, Zophobas morio λmax=613 nm), and mutants of C311 (S314). The green-yellow-emitting luciferases display red-shifted bioluminescence spectra with aminoluciferin in relation to those with D-luciferin, whereas the red-emitting luciferases displayed blue-shifted spectra. Bioluminescence spectra with 5,5-dimethylaminoluciferin, in which enolization is blocked, were almost identical to those of aminoluciferin. Fluorescence probing using 2-(4-toluidino)naphthalene-6-sulfonate and inference with aminoluciferin confirm that the luciferin binding site of the red-shifted luciferases is more polar than in the case of the green-yellow-emitting luciferases. Altogether, the results show that the keto form of excited oxyluciferin is the emitter in beetle bioluminescence and that bioluminescence colors are essentially modulated by interactions of the 6'-hydroxy group of oxyluciferin and basic moieties under the influence of the microenvironment polarity of the active site: a strong interaction between a base moiety and oxyluciferin phenol in a hydrophobic microenvironment promotes green-yellow emission, whereas a more polar

  9. Processes influencing the transport and fate of contaminated sediments in the coastal ocean: Boston Harbor and Massachusetts Bay

    Science.gov (United States)

    Alexander, P. Soupy; Baldwin, Sandra M.; Blackwood, Dann S.; Borden, Jonathan; Casso, Michael A.; Crusius, John; Goudreau, Joanne; Kalnejais, Linda H.; Lamothe, Paul J.; Martin, William R.; Martini, Marinna A.; Rendigs, Richard R.; Sayles, Frederick L.; Signell, Richard P.; Valentine, Page C.; Warner, John C.; Bothner, Michael H.; Butman, Bradford

    2007-01-01

    Most of the major urban centers of the United States including Boston, New York, Washington, Chicago, New Orleans, Miami, Los Angeles, San Francisco, and Seattle—are on a coast (fig. 1.1). All of these cities discharge treated sewage effluent into adjacent waters. In 2000, 74 percent of the U.S. population lived within 200 kilometers (km) of the coast. Between 1980 and 2002, the population density in coastal communities increased approximately 4.5 times faster than in noncoastal areas of the U.S. (Perkins, 2004). More people generate larger volumes of wastes, increase the demands on wastewater treatment, expand the area of impervious land surfaces, and use more vehicles that contribute contaminants to street runoff. According to the National Coastal Condition Report II (U.S. Environmental Protection Agency, 2005a), on the basis of coastal habitat, water and sediment quality, benthic index, and fish tissue, the overall national coastal condition is only poor to fair and the overall coastal condition in the highly populated Northeast is poor. Scientific information helps managers to prioritize and regulate coastal-ocean uses that include recreation, commercial fishing, transportation, waste disposal, and critical habitat for marine organisms. These uses are often in conflict with each other and with environmental concerns. Developing a strategy for managing competing uses while maintaining sustainability of coastal resources requires scientific understanding of how the coastal ocean system behaves and how it responds to anthropogenic influences. This report provides a summary of a multidisciplinary research program designed to improve our understanding of the transport and fate of contaminants in Massachusetts coastal waters. Massachusetts Bay and Boston Harbor have been a focus of U.S. Geological Survey (USGS) research because they provide a diverse geographic setting for developing a scientific understanding of the geology, geochemistry, and oceanography of

  10. Ecological evaluation of proposed dredged material from Richmond Harbor Deepening Project and the intensive study of the Turning Basin

    Energy Technology Data Exchange (ETDEWEB)

    Pinza, M.R.; Mayhew, H.L.; Karle, L.M.; Kohn, N.P.; White, P.J.; Word, J.Q.; Michaels, L.L. [Battelle/Marine Sciences Lab., Sequim, WA (United States)

    1995-06-01

    Richmond Harbor is on the eastern shoreline of central San Francisco Bay and its access channels and several of the shipping berths are no longer wide or deep enough to accommodate modem deeper-draft vessels. The Water Resources Development Act of 1986 (PL99-662) authorized the US Army Corps of Engineers (USACE), San Francisco District to deepen and widen the navigation channels in Richmond Harbor. Several options for disposal of the material from this dredging project are under consideration by USACE: disposal within San Francisco Bay, at open-ocean disposal sites, or at uplands disposal sites. Purpose of this study was to conduct comprehensive evaluations, including chemical, biological, and bioaccumulation testing of sediments in selected areas of Richmond Harbor. This information was required by the Environmental Protection Agency (EPA) and USACE. Battelle/Marine Sciences Laboratory collected 20 core samples, both 4-in. and 12-in., to a project depth of -40 ft mean lower low water (MLLW) (-38 ft MLLW plus 2 ft of overdepth) using a vibratory-hammer core. These 20 field samples were combined to form five test composites plus an older bay mud (OBM) composite that were analyzed for physical/chemical parameters, biological toxicity, and tissue chemistry. Solid-phase tests were conducted with the amphipod, Rhepoxynius abronius; the clam, Macoma nasuta; and the polychaete worm, Nephtys caecoides. Suspended-particulate-phase (SPP) tests were conducted with the sanddab, Citharichthys stigmaeus; the mysid, Holmesimysis costata; and the bivalve, Mytilus galloprovincialis. Bioaccumulation of contaminants was measured in tissues of Macoma nasuta and Nereis virens. Sediments from one ocean reference sediment, and two in-bay reference sediments, were tested concurrently. Results from analysis of the five test treatments were statistically compared with the reference sediment R-OS in the first five sections of this report.

  11. Mnemiopsis leidyi Gut Harbors Seasonally Variant and Commensal Microbial Assemblages

    Science.gov (United States)

    Mariita, R. M.; Hossain, M. J.; Liles, M. R.; Moss, A.

    2016-02-01

    Studies have shown that with widespread use of antibiotics in human and domestic animal populations, antibiotic resistance becomes increasingly common in the environment. Estuaries provide ideal conditions for acquisition and dissemination of drug resistance genes because they serve as sinks for pollution. This study aimed to identify M. leidyi microbial diversity and richness and their potential to act as vectors for antibiotic resistance determinants (ARDs). M. leidyi, although native to study area are highly invasive. Metagenomic analyses indicate that there are temporal variations of bacterioplankton assemblages in M. leidyi gut. Overall, Proteobacteria and Actinobacteria are the most abundant phyla. Despite the temporal dynamics in the microbial assemblages in M. leidyi gut, they seem to retain Propionibacterium acnes (gut microbiota in some insects) and select proteobacteria across all seasons. The results contradict previous studies that suggest that M. leidyi does not have constant a microbiota, but only seasonally variant microbial assemblages. Here we reveal the presence of M. leidyi gut ARDs in winter and summer, probably because of the ctenophores' positive geotaxis during rough surface conditions. Genes responsible for resistance to fluoroquinolones, multidrug resistance efflux pumps, mercuric reductase, copper homeostasis and blaR1 genes were observed. This is the first study to demonstrate that M. leidyi harbors constant microbiota and provides a baseline for understanding M. leidyi gut microbial and ARDs ecology. It also suggests that M. leidyi bacterial taxonomic and functional dynamics is influenced by season. Funding: Alabama EPSCoR GRSP fellowship, AU-CMB fellowship, NSF EPS-1158862, USDA-Hatch 370225-310100 (AGM, ML).

  12. Settlement of the USS Arizona, Pearl Harbor, Hawaii

    Science.gov (United States)

    Carkin, Brad A.; Kayen, Robert E.

    2013-01-01

    The U.S. Geological Survey, in collaboration with the National Park Service Submerged Resources Center, undertook investigations at the USS Arizona Memorial at Pearl Harbor, Hawaii, in 2002, 2003, and 2005 to characterize geological factors affecting the deterioration and movement of the hull of the USS Arizona. Since sinking on the morning of December 7, 1941, the hull of the USS Arizona has been slowly but steadily disappearing below the surface of Pearl Harbor. Continuous sediment coring at three of four locations around the hull of the Arizona was only partially successful, but it was sufficient to identify a varied sedimentary substrate beneath the hull. A boring near the stern reveals a thick, continuous sequence of soft, gray clay to the bottom of the boring. In contrast, borings near the bow and starboard side, below about 5 meters subbottom depth, indicate the presence of very stiff, brown clay and coral debris and an absence of soft clay. Multisensor core logger scanning of the recovered cores distinguishes the lower density of the soft, gray clay at the stern from the higher density of the stiff, brown clays and coral debris at the bow and starboard side. Uniaxial consolidation testing of the soft gray clay indicates a normally consolidated sequence, whereas the stiff, brown clay and coral debris are overconsolidated. Profiles of shear wave velocity vs. depth obtained through spectral analysis of interface wave testing around the perimeter of the hull in 2005 identified areas of higher velocity, stiffer sediment at the bow and starboard side, which correspond to the dense, stiff clay recovered near the bow and starboard borings. Low shear-wave velocities at the port midship and quarter of the hull correlate with the lower density, softer sediment recovered from the boring at the stern. Cross sections of the subbottom of the Memorial combine results from the sediment borings and geophysical surveys and depict a wedge of soft clay unconformably overlying

  13. Onondaga Lake Inner Harbor Dredging Design Project, Syracuse, New York: Final Design Memorandum

    National Research Council Canada - National Science Library

    1997-01-01

    .... The sponsor is the New York State Canal Corporation. The design includes deepening the Inner Harbor channel and a portion of the terminal slip area to a depth of 10 feet below Low Water Datum (LWD...

  14. Aerial Survey Trend Counts of Harbor Seals in Coastal Alaska (1984-2006) - ADF&G

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Aerial surveys were conducted during 1983–2006 in the Ketchikan, Sitka, Kodiak, and Bristol Bay areas of Alaska to estimate trends in abundance of harbor seals.

  15. Aerial Survey Counts of Harbor Seals in Lake Iliamna, Alaska, 1984-2013 (NODC Accession 0123188)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This dataset provides counts of harbor seals from aerial surveys over Lake Iliamna, Alaska, USA. The data have been collated from three previously published sources...

  16. Sedimentation and chronology of heavy metal pollution in Oslo harbor, Norway

    International Nuclear Information System (INIS)

    Lepland, Aivo; Andersen, Thorbjorn J.; Lepland, Aave; Arp, Hans Peter H.; Alve, Elisabeth; Breedveld, Gijs D.; Rindby, Anders

    2010-01-01

    Stratigraphic profiles of Cu, Cd and Hg in ten sediment cores from the Oslo harbor, Norway, combined with results of radiometric dating demonstrate that pollution by these metals peaked between 1940 and 1970. Dating results indicate that Hg discharges peaked between 1940 and 1950, Cd reached maximum ca. 1955-1960, and Cu has the highest concentration in sediment interval corresponding to ca. 1970. Geochemical profiles and maxima of Cu, Cd and Hg concentrations can be used as chronostratigraphic markers for sediment cores from the Oslo harbor. Acoustic backscatter and sediment core data indicate that propeller wash affects the seabed in the Oslo harbor. The propeller-induced turbulence causes erosion, and in places exposes and remobilizes contaminated sediments that accumulated in the harbor during previous decades. Such re-exposure of contaminated sediments could be detrimental to local ecosystems and offset remediation efforts, warranting further impact studies and potential mitigation strategies to prevent redistribution.

  17. Wave Climate and Wave Response, Kawaihae Deep Draft Harbor, Island of Hawaii, Hawaii

    National Research Council Canada - National Science Library

    Thompson, Edward F; Demirbilek, Zeki; Briggs, Michael J

    2006-01-01

    Present and projected commercial activities in Kawaihae Deep Draft Harbor, Island of Hawaii, HI, indicate that a deeper basin and entrance channel and better protected berthing areas will be needed. The U.S...

  18. Inshore Survey Results: Approaches to New York Harbor, Fall 1955 (NODC Accession 7000294)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — A current survey in the approaches to New York Harbor was initiated 06 September 1955 and continued through 04 November 1955. This survey was conducted in accordance...

  19. Environmental Assessment for Building 88 Interior Demolition, Pearl Harbor Naval Complex, O'ahu, Hawaii

    National Research Council Canada - National Science Library

    2005-01-01

    ...) and ancillary equipment from the interior of Building 88, a former lubricating oil storage facility, and the related exposed piping underneath Mike wharves M3 and M4 at Merry Point, Pearl Harbor Naval Complex...

  20. Defense Infrastructure: General and Flag Officer Quarters at Pearl Harbor, Hawaii

    National Research Council Canada - National Science Library

    2002-01-01

    .... We reviewed 17 GFOQs at Pearl Harbor, Hawaii, with budgeted maintenance and repair costs of $1,247,300, to determine whether the Navy had properly classified interior shutter costs as maintenance and repair...

  1. Environmental Assessment for Waterfront Facilities Maintenance and Improvements, Pearl Harbor Naval Complex, Oahu, Hawaii

    National Research Council Canada - National Science Library

    2005-01-01

    Commander, Navy Region Hawaii (CNRH) proposes to repair, maintain, and improve waterfront berthing and maintenance facilities for ships and submarines on an as-needed basis within the Pearl Harbor Naval Complex (PHNC...

  2. Gridded multibeam bathymetry of Apra Harbor, Guam U.S. Territory

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Gridded bathymetry from Apra Harbor, Guam U.S. Territory. The netCDF and Arc ASCII grids include multibeam bathymetry from the Reson SeaBat 8125 multibeam sonar...

  3. DefenseLink Special: Attack on Pearl Harbor, 65th Anniversary

    Science.gov (United States)

    Japanese raid on Pearl Harbor has been called a defining moment in U.S. history. It caught the country by Hawaii-based combat planes, were heavily damaged. By crippling the U.S. Pacific Fleet, Japan hoped to

  4. MODELING HOW A HURRICANE BARRIER IN NEW BEDFORD HARBOR, MASSACHUSETTS, AFFECTS THE HYDRODYNAMICS AND RESIDENCE TIMES

    Science.gov (United States)

    Two-dimensional hydrodynamic and transport models were used to simulate tidal and subtidal circulation, residence times, and the longitudinal distributions of conservative constituents in New Bedford Harbor, Massachusetts, before and after a hurricane barrier was constructed. The...

  5. Aerial Survey Effort for Harbor Seals in Coastal Alaska (2004-2015)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The most feasible approach to determining harbor seal distribution and abundance in Alaska coastal habitats is to use aircraft to count seals when they haul out of...

  6. A Dataset of Aerial Survey Counts of Harbor Seals in Iliamna Lake, Alaska: 1984-2013

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This dataset provides counts of harbor seals from aerial surveys over Iliamna Lake, Alaska, USA. The data have been collated from three previously published sources...

  7. Ice Harbor Spillway Dissolved Gas Field Studies: Before and After Spillway Deflectors

    Science.gov (United States)

    2016-07-01

    Steven C. Wilhelms Coastal And Hydraulics Laboratory U.S. Army Engineer Research and Development Center 3909 Halls Ferry Road Vicksburg, Mississippi...Harbor as a measure to reduce the total dissolved gas (TDG) production during spill operations. Three field studies were conducted at the Ice Harbor...significantly reduced for nearly all spill operations with deflectors in place. TDG near the stilling basin was reduced from approximately 150% to

  8. Analysis of Protection Measures for Naval Vessels Berthed at Harbor Against Terrorist Attacks

    Science.gov (United States)

    2016-06-01

    of discriminating neutral vessels from threats. A naval vessel berthed at harbor is more susceptible to attack than a vessel in open seas. The...discriminating neutral vessels from threats. A naval vessel berthed at harbor is more susceptible to attack than a vessel in open seas. The chances of...this thesis. He was a source of inspiration, encouragement, and reassurance. Captain Jeffery E. Kline, I am really thankful to you for your ideas

  9. Cladophora (Chlorophyta) spp. Harbor Human Bacterial Pathogens in Nearshore Water of Lake Michigan†

    OpenAIRE

    Ishii, Satoshi; Yan, Tao; Shively, Dawn A.; Byappanahalli, Muruleedhara N.; Whitman, Richard L.; Sadowsky, Michael J.

    2006-01-01

    Cladophora glomerata, a macrophytic green alga, is commonly found in the Great Lakes, and significant accumulations occur along shorelines during the summer months. Recently, Cladophora has been shown to harbor high densities of the fecal indicator bacteria Escherichia coli and enterococci. Cladophora may also harbor human pathogens; however, until now, no studies to address this question have been performed. In the present study, we determined whether attached Cladophora, obtained from the L...

  10. 75 FR 43823 - Safety Zone; He'eia Kea Small Boat Harbor, Kaneohe Bay, Oahu, HI

    Science.gov (United States)

    2010-07-27

    ...The Coast Guard is establishing a temporary safety zone in He'eia Kea Small Boat Harbor located in Kaneohe Bay, Oahu, Hawaii. The safety zone is necessary to protect watercraft and the general public from hazards associated with five vessels moored for approximately 3- weeks off the boat harbor's main pier. Vessels desiring to transit through the zone can request permission by contacting the Captain of the Port Honolulu.

  11. A solitary hyperfunctioning thyroid nodule harboring thyroid carcinoma: review of the literature

    OpenAIRE

    Mirfakhraee, Sasan; Mathews, Dana; Peng, Lan; Woodruff, Stacey; Zigman, Jeffrey M

    2013-01-01

    Hyperfunctioning nodules of the thyroid are thought to only rarely harbor thyroid cancer, and thus are infrequently biopsied. Here, we present the case of a patient with a hyperfunctioning thyroid nodule harboring thyroid carcinoma and, using MEDLINE literature searches, set out to determine the prevalence of and characteristics of malignant ?hot? nodules as a group. Historical, biochemical and radiologic characteristics of the case subjects and their nodules were compared to those in cases o...

  12. Numerical Modeling of Wave Overtopping of Buffalo Harbor Confined Disposal Facility (CDF4)

    Science.gov (United States)

    2017-10-01

    navigation channel , and harbor complex. Though there are anecdotal claims and debris indicating possible sediment movement, there has been no evidence of...littoral sediments inside the disposal area, into the channel and harbor, and other adjacent areas to the CDF4. 1.4 Study plan 1.4.1 Purpose There...locations. 2.4 Maintenance dredging data The need for maintenance dredging arises from sedimentation in the navigation channels , which impedes

  13. Impact of Geomorphological Changes to Harbor Resonance During Meteotsunamis: The Vela Luka Bay Test Case

    Science.gov (United States)

    Denamiel, Cléa; Šepić, Jadranka; Vilibić, Ivica

    2018-05-01

    In engineering studies, harbor resonance, including quality and amplification factors, is typically computed for swell and waves with periods shorter than 10 min. However, in various locations around the world, such as Vela Luka Bay in Croatia, meteotsunami waves of periods greater than 10 min can excite the bay or harbor natural modes and produce substantial structural damages. In this theoretical study, the impact of some geomorphological changes of Vela Luka Bay—i.e. deepening of the bay, dredging the harbor, adding a pier or a marina—to the amplification of the meteotsunami waves are presented for a set of 6401 idealized pressure wave field forcing used to derive robust statistics. The most substantial increase in maximum elevation is found when the Vela Luka harbor is dredged to a 5 m depth, which is in contradiction with the calculation of the quality factor showing a decrease of the harbor natural resonance. It has been shown that the forcing energy content at different frequency bands should also be taken into account when estimating the quality and amplification factors, as their typical definitions derived from the peak frequency of the sea level spectrum fail to represent the harbor response during meteotsunami events. New definitions of these factors are proposed in this study and are shown to be in good agreement with the results of the statistical analysis of the Vela Luka Bay maximum elevation results. In addition, the presented methodology can easily be applicable to any other location in the world where meteotsunamis occur.

  14. High bacterial biodiversity increases degradation performance of hydrocarbons during bioremediation of contaminated harbor marine sediments

    International Nuclear Information System (INIS)

    Dell'Anno, Antonio; Beolchini, Francesca; Rocchetti, Laura; Luna, Gian Marco; Danovaro, Roberto

    2012-01-01

    We investigated changes of bacterial abundance and biodiversity during bioremediation experiments carried out on oxic and anoxic marine harbor sediments contaminated with hydrocarbons. Oxic sediments, supplied with inorganic nutrients, were incubated in aerobic conditions at 20 °C and 35 °C for 30 days, whereas anoxic sediments, amended with organic substrates, were incubated in anaerobic conditions at the same temperatures for 60 days. Results reported here indicate that temperature exerted the main effect on bacterial abundance, diversity and assemblage composition. At higher temperature bacterial diversity and evenness increased significantly in aerobic conditions, whilst decreased in anaerobic conditions. In both aerobic and anaerobic conditions, biodegradation efficiencies of hydrocarbons were significantly and positively related with bacterial richness and evenness. Overall results presented here suggest that bioremediation strategies, which can sustain high levels of bacterial diversity rather than the selection of specific taxa, may significantly increase the efficiency of hydrocarbon degradation in contaminated marine sediments. - Highlights: ► Bioremediation performance was investigated on hydrocarbon contaminated sediments. ► Major changes in bacterial diversity and assemblage composition were observed. ► Temperature exerted the major effect on bacterial assemblages. ► High bacterial diversity increased significantly biodegradation performance. ► This should be considered for sediment remediation by bio-treatments. - Bioremediation strategies which can sustain high levels of bacterial diversity may significantly increase the biodegradation of hydrocarbons in contaminated marine sediments.

  15. Virulence of Klebsiella pneumoniae isolates harboring bla KPC-2 carbapenemase gene in a Caenorhabditis elegans model.

    Directory of Open Access Journals (Sweden)

    Jean-Philippe Lavigne

    Full Text Available Klebsiella pneumoniae carbapenemase (KPC is a carbapenemase increasingly reported worldwide in Enterobacteriaceae. The aim of this study was to analyze the virulence of several KPC-2-producing K. pneumoniae isolates. The studied strains were (i five KPC-2 clinical strains from different geographical origins, belonging to different ST-types and possessing plasmids of different incompatibility groups; (ii seven transformants obtained after electroporation of either these natural KPC plasmids or a recombinant plasmid harboring only the bla KPC-2 gene into reference strains K. pneumoniae ATCC10031/CIP53153; and (iii five clinical strains cured of plasmids. The virulence of K. pneumoniae isolates was evaluated in the Caenorhabditis elegans model. The clinical KPC producers and transformants were significantly less virulent (LT50: 5.5 days than K. pneumoniae reference strain (LT50: 4.3 days (p<0.01. However, the worldwide spread KPC-2 positive K. pneumoniae ST258 strains and reference strains containing plasmids extracted from K. pneumoniae ST258 strains had a higher virulence than KPC-2 strains belonging to other ST types (LT50: 5 days vs. 6 days, p<0.01. The increased virulence observed in cured strains confirmed this trend. The bla KPC-2 gene itself was not associated to increased virulence.

  16. Beyond Parasitism: Hepatic Lesions in Stranded Harbor Porpoises (Phocoena phocoena) Without Trematode (Campula oblonga) Infections.

    Science.gov (United States)

    Hiemstra, S; Harkema, L; Wiersma, L C M; Keesler, R I

    2015-11-01

    The liver can be an indicator of the health of an individual or of a group, which can be especially important to identify agents that can cause disease in multiple species. To better characterize hepatic lesions in stranded harbor porpoises (Phocoena phocoena), we analyzed the livers from 39 porpoises that stranded along the Dutch coast between December 2008 and December 2012. The animals were selected because they had either gross or histologic liver lesions with minimal autolysis and no evidence of trematode (Campula oblonga) infection. The most common finding was a chronic hepatitis (22/39, 56.4%) that was often associated with significant disease reported in another organ system (18/22, 81.8%), of which 14 had chronic systemic disease. One case of chronic hepatitis was so severe as to mimic lymphoma, which could only be differentiated with immunohistochemistry. The other common lesions were lipidosis (11/39, 28.2%) and acute hepatitis (6/39, 15.4%), often in combination with mild chronic changes. Overall, although there were no consistent trends in etiology for the hepatic lesions, lipidosis was associated with starvation (8/11, 72.7%) and acute disease, and acute hepatitis was associated with bacterial infections and sepsis (6/6, 100%). © The Author(s) 2014.

  17. Sequential Kinase Inhibition (Idelalisib/Ibrutinib Induces Clinical Remission in B-Cell Prolymphocytic Leukemia Harboring a 17p Deletion

    Directory of Open Access Journals (Sweden)

    H. Coelho

    2017-01-01

    Full Text Available B-cell prolymphocytic leukemia (B-PLL is a rare lymphoid neoplasm with an aggressive clinical course. Treatment strategies for B-PLL remain to be established, and, until recently, alemtuzumab was the only effective therapeutic option in patients harboring 17p deletions. Herein, we describe, for the first time, a case of B-cell prolymphocytic leukemia harboring a 17p deletion in a 48-year-old man that was successfully treated sequentially with idelalisib-rituximab/ibrutinib followed by allogeneic hematopoietic stem cell transplant (allo-HSCT. After 5 months of therapy with idelalisib-rituximab, clinical remission was achieved, but the development of severe diarrhea led to its discontinuation. Subsequently, the patient was treated for 2 months with ibrutinib and the quality of the response was maintained with no severe adverse effects reported. A reduced-intensity conditioning allo-HSCT from a HLA-matched unrelated donor was performed, and, thereafter, the patient has been in complete remission for 10 months now. In conclusion, given the poor prognosis of B-PLL and the lack of effective treatment modalities, the findings here suggest that both ibrutinib and idelalisib should be considered as upfront therapy of B-PLL and as a bridge to allo-HSCT.

  18. Metastasizing, Luciferase Transduced MAT-Lu Rat Prostate Cancer Models: Follow up of Bolus and Metronomic Therapy with Doxorubicin as Model Drug

    International Nuclear Information System (INIS)

    Jantscheff, Peter; Esser, Norbert; Geipel, Andreas; Woias, Peter; Ziroli, Vittorio; Goldschmidtboing, Frank; Massing, Ulrich

    2011-01-01

    The most fatal outcomes of prostate carcinoma (PCa) result from hormone-refractory variants of the tumor, especially from metastatic spread rather than from primary tumor burden. The goal of the study was to establish and apply rat MAT-Lu prostate cancer tumor models for improved non-invasive live follow up of tumor growth and metastasis by in vivo bioluminescence. We established luciferase transduced MAT-Lu rat PCa cells and studied tumor growth and metastatic processes in an ectopic as well as orthotopic setting. An intravenous bolus treatment with doxorubicin was used to demonstrate the basic applicability of in vivo imaging to follow up therapeutic intervention in these models. In vitro analysis of tissue homogenates confirmed major metastatic spread of subcutaneous tumors into the lung. Our sensitive method, however, for the first time detects metastasis also in lymph node (11/24), spleen (3/24), kidney (4/24), liver (5/24), and bone tissue (femur or spinal cord - 5/20 and 12/20, respectively). Preliminary data of orthotopic implantation (three animals) showed metastatic invasion to investigated organs in all animals but with varying preference (e.g., to lymph nodes). Intravenous bolus treatment of MAT-Lu PCa with doxorubicin reduced subcutaneous tumor growth by about 50% and the number of animals affected by metastatic lesions in lymph nodes (0/4), lung (3/6) or lumbar spine (0/2), as determined by in vivo imaging and in vitro analysis. Additionally, the possible applicability of the luciferase transduced MAT-Lu model(s) to study basic principles of metronomic therapies via jugular vein catheter, using newly established active microport pumping systems, is presented

  19. Metastasizing, Luciferase Transduced MAT-Lu Rat Prostate Cancer Models: Follow up of Bolus and Metronomic Therapy with Doxorubicin as Model Drug

    Energy Technology Data Exchange (ETDEWEB)

    Jantscheff, Peter, E-mail: jantscheff@tumorbio.uni-freiburg.de [Tumour Biology Center, Clinical Research, Department Lipids & Liposomes, Breisacher Str.117, D-79106 Freiburg (Germany); Esser, Norbert [ProQinase GmbH, Breisacher Str. 117, D-79106 Freiburg (Germany); Geipel, Andreas; Woias, Peter [Laboratory for Design of Microsystems, Department of Microsystems Engineering (IMTEK), Georges-Köhler-Allee 106, D-79110 Freiburg (Germany); Ziroli, Vittorio [Tumour Biology Center, Clinical Research, Department Lipids & Liposomes, Breisacher Str.117, D-79106 Freiburg (Germany); Goldschmidtboing, Frank [Laboratory for Design of Microsystems, Department of Microsystems Engineering (IMTEK), Georges-Köhler-Allee 106, D-79110 Freiburg (Germany); Massing, Ulrich [Tumour Biology Center, Clinical Research, Department Lipids & Liposomes, Breisacher Str.117, D-79106 Freiburg (Germany)

    2011-06-17

    The most fatal outcomes of prostate carcinoma (PCa) result from hormone-refractory variants of the tumor, especially from metastatic spread rather than from primary tumor burden. The goal of the study was to establish and apply rat MAT-Lu prostate cancer tumor models for improved non-invasive live follow up of tumor growth and metastasis by in vivo bioluminescence. We established luciferase transduced MAT-Lu rat PCa cells and studied tumor growth and metastatic processes in an ectopic as well as orthotopic setting. An intravenous bolus treatment with doxorubicin was used to demonstrate the basic applicability of in vivo imaging to follow up therapeutic intervention in these models. In vitro analysis of tissue homogenates confirmed major metastatic spread of subcutaneous tumors into the lung. Our sensitive method, however, for the first time detects metastasis also in lymph node (11/24), spleen (3/24), kidney (4/24), liver (5/24), and bone tissue (femur or spinal cord - 5/20 and 12/20, respectively). Preliminary data of orthotopic implantation (three animals) showed metastatic invasion to investigated organs in all animals but with varying preference (e.g., to lymph nodes). Intravenous bolus treatment of MAT-Lu PCa with doxorubicin reduced subcutaneous tumor growth by about 50% and the number of animals affected by metastatic lesions in lymph nodes (0/4), lung (3/6) or lumbar spine (0/2), as determined by in vivo imaging and in vitro analysis. Additionally, the possible applicability of the luciferase transduced MAT-Lu model(s) to study basic principles of metronomic therapies via jugular vein catheter, using newly established active microport pumping systems, is presented.

  20. Metastasizing, Luciferase Transduced MAT‑Lu Rat Prostate Cancer Models: Follow up of Bolus and Metronomic Therapy with Doxorubicin as Model Drug

    Directory of Open Access Journals (Sweden)

    Peter Woias

    2011-06-01

    Full Text Available The most fatal outcomes of prostate carcinoma (PCa result from hormone-refractory variants of the tumor, especially from metastatic spread rather than from primary tumor burden. The goal of the study was to establish and apply rat MAT-Lu prostate cancer tumor models for improved non-invasive live follow up of tumor growth and metastasis by in vivo bioluminescence. We established luciferase transduced MAT-Lu rat PCa cells and studied tumor growth and metastatic processes in an ectopic as well as orthotopic setting. An intravenous bolus treatment with doxorubicin was used to demonstrate the basic applicability of in vivo imaging to follow up therapeutic intervention in these models. In vitro analysis of tissue homogenates confirmed major metastatic spread of subcutaneous tumors into the lung. Our sensitive method, however, for the first time detects metastasis also in lymph node (11/24, spleen (3/24, kidney (4/24, liver (5/24, and bone tissue (femur or spinal cord - 5/20 and 12/20, respectively. Preliminary data of orthotopic implantation (three animals showed metastatic invasion to investigated organs in all animals but with varying preference (e.g., to lymph nodes. Intravenous bolus treatment of MAT-Lu PCa with doxorubicin reduced subcutaneous tumor growth by about 50% and the number of animals affected by metastatic lesions in lymph nodes (0/4, lung (3/6 or lumbar spine (0/2, as determined by in vivo imaging and in vitro analysis. Additionally, the possible applicability of the luciferase transduced MAT-Lu model(s to study basic principles of metronomic therapies via jugular vein catheter, using newly established active microport pumping systems, is presented.

  1. Disruption of bbe02 by Insertion of a Luciferase Gene Increases Transformation Efficiency of Borrelia burgdorferi and Allows Live Imaging in Lyme Disease Susceptible C3H Mice.

    Directory of Open Access Journals (Sweden)

    Kamfai Chan

    Full Text Available Lyme disease is the most prevalent tick-borne disease in North America and Europe. The causative agent, Borrelia burgdorferi persists in the white-footed mouse. Infection with B. burgdorferi can cause acute to persistent multisystemic Lyme disease in humans. Some disease manifestations are also exhibited in the mouse model of Lyme disease. Genetic manipulation of B. burgdorferi remains difficult. First, B. burgdorferi contains a large number of endogenous plasmids with unique sequences encoding unknown functions. The presence of these plasmids needs to be confirmed after each genetic manipulation. Second, the restriction modification defense systems, including that encoded by bbe02 gene lead to low transformation efficiency in B. burgdorferi. Therefore, studying the molecular basis of Lyme pathogenesis is a challenge. Furthermore, investigation of the role of a specific B. burgdorferi protein throughout infection requires a large number of mice, making it labor intensive and expensive. To overcome the problems associated with low transformation efficiency and to reduce the number of mice needed for experiments, we disrupted the bbe02 gene of a highly infectious and pathogenic B. burgdorferi strain, N40 D10/E9 through insertion of a firefly luciferase gene. The bbe02 mutant shows higher transformation efficiency and maintains luciferase activity throughout infection as detected by live imaging of mice. Infectivity and pathogenesis of this mutant were comparable to the wild-type N40 strain. This mutant will serve as an ideal parental strain to examine the roles of various B. burgdorferi proteins in Lyme pathogenesis in the mouse model in the future.

  2. Inhibition of metastatic tumor growth in mouse lung by repeated administration of polyethylene glycol-conjugated catalase: quantitative analysis with firefly luciferase-expressing melanoma cells.

    Science.gov (United States)

    Hyoudou, Kenji; Nishikawa, Makiya; Umeyama, Yukari; Kobayashi, Yuki; Yamashita, Fumiyoshi; Hashida, Mitsuru

    2004-11-15

    To develop a novel and effective approach to inhibit tumor metastasis based on controlled delivery of catalase, we first evaluated the characteristics of the disposition and proliferation of tumor cells. Then, we examined the effects of polyethylene glycol-conjugated catalase (PEG-catalase) on tumor metastasis. On the basis of the results obtained, PEG-catalase was repetitively administered to completely suppress the growth of tumor cells. Murine melanoma B16-BL6 cells were stably transfected with firefly luciferase gene to obtain B16-BL6/Luc cells. These cells were injected intravenously into syngeneic C57BL/6 mice. PEG-catalase was injected intravenously, and the effect was evaluated by measuring the luciferase activity as the indicator of the number of tumor cells. At 1 hour after injection of B16-BL6/Luc cells, 60 to 90% of the injected cells were recovered in the lung. The numbers decreased to 2 to 4% at 24 hours, then increased. An injection of PEG-catalase just before inoculation significantly reduced the number of tumor cells at 24 hours. Injection of PEG-catalase at 1 or 3 days after inoculation was also effective in reducing the cell numbers. Daily dosing of PEG-catalase greatly inhibited the proliferation and the number assayed at 14 days after inoculation was not significantly different from the minimal number observed at 1 day, suggesting that the growth had been markedly suppressed by the treatment. These findings indicate that sustained catalase activity in the blood circulation can prevent the multiple processes of tumor metastasis in the lung, which could lead to a state of tumor dormancy.

  3. Copper storage in the liver of the wild mute swan (Cygnus olor). Its possible relation to pollution of harbor waters by antifouling paints.

    Science.gov (United States)

    Molnar, J J

    1983-12-01

    Postmortem examination of three wild mute swans (Cygnus olor) from a harbor area disclosed an unusual black discoloration of the liver. Chemical, histochemical, and microscopic studies, along with electron-probe microanalysis, showed that cytoplasmic pigment granules in the liver cells contained a copper-protein complex. Similar findings have been reported in Danish and English studies on large numbers of wild mute swans. Two control mute swans from The Bronx Zoo had negligible amounts of hepatic copper. The striking difference between the wild and the captive swans in hepatic copper content suggests that the copper in the wild swans was of environmental origin, most likely from copper-rich antifouling paint used extensively in the marine industry. Flakes of this paint may be ingested by swans searching for food in the sediment of harbor waters.

  4. Environmental gamma radiation measurements over the Bay of Bengal around Kalpakkam and in Chennai Harbor

    International Nuclear Information System (INIS)

    Surya Prakash, G.; Baskaran, R.; Venkatraman, B.; Rajagopal, V.; Mohanty, Ajit Kumar

    2012-01-01

    DAE site Kalpakkam, encompassing several operating nuclear facilitates is enclosed in a strip of land between the Bay of Bengal on the east and Buckingham Canal on the west. Approximately about 50% of the Emergency Prone Zone (EPZ) area around the plant lies in the Bay of Bengal. The radiation levels around Kalpakkam nuclear complex is monitored by Environmental Survey Laboratory (ESL) at Kalpakkam. The survey has been carried out up to 16 km in the land areas. Though radioactivity levels in the seawater and the food produce derived from the sea are monitored and reported by ESL, data on the ambient radiation levels over the sea has not been reported. The paper describes the campaign based ambient gamma radiation level measurements done during the past few years. The radiation level measurements done at selected locations varied between 8 and 115 nGyh -1 (0.8 and 11.5 μRh -1 ) during the first campaign and 14 and 170 nGyh -1 (1.4 and 17 μRh -1 ) during the second campaign. The average dose rate observed from both the campaigns was 62 nGyh 1 (6.2 μRh -1 ). During the campaign period we are looked in to any interference from the 41 Ar released from the MAPS reactor also. The third campaign at the Chennai harbor (∼ 60 km north of Kalpakkam) the observed radiation levels were between 15 nGyh -1 (1.5 μRh -1 ) and 12.2 nGyh -1 (12.2 μRh -1 ). This paper discussed the details of the techniques and the measurement

  5. Specific and Efficient Regression of Cancers Harboring KRAS Mutation by Targeted RNA Replacement.

    Science.gov (United States)

    Kim, Sung Jin; Kim, Ju Hyun; Yang, Bitna; Jeong, Jin-Sook; Lee, Seong-Wook

    2017-02-01

    Mutations in the KRAS gene, which persistently activate RAS function, are most frequently found in many types of human cancers. Here, we proposed and verified a new approach against cancers harboring the KRAS mutation with high cancer selectivity and efficient anti-cancer effects based on targeted RNA replacement. To this end, trans-splicing ribozymes from Tetrahymena group I intron were developed, which can specifically target and reprogram the mutant KRAS G12V transcript to induce therapeutic gene activity in cells. Adenoviral vectors containing the specific ribozymes with downstream suicide gene were constructed and then infection with the adenoviruses specifically downregulated KRAS G12V expression and killed KRAS G12V-harboring cancer cells additively upon pro-drug treatment, but it did not affect the growth of wild-type KRAS-expressing cells. Minimal liver toxicity was noted when the adenoviruses were administered systemically in vivo. Importantly, intratumoral injection of the adenoviruses with pro-drug treatment specifically and significantly impeded the growth of xenografted tumors harboring KRAS G12V through a trans-splicing reaction with the target RNA. In contrast, xenografted tumors harboring wild-type KRAS were not affected by the adenoviruses. Therefore, RNA replacement with a mutant KRAS-targeting trans-splicing ribozyme is a potentially useful therapeutic strategy to combat tumors harboring KRAS mutation. Copyright © 2017 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.

  6. Reducing Vulnerability of Ports and Harbors to Earthquake and Tsunami Hazards

    Science.gov (United States)

    Wood, Nathan J.; Good, James W.; Goodwin, Robert F.

    2002-01-01

    Recent scientific research suggests the Pacific Northwest could experience catastrophic earthquakes in the near future, both from distant and local sources, posing a significant threat to coastal communities. Damage could result from numerous earthquake-related hazards, such as severe ground shaking, soil liquefaction, landslides, land subsidence/uplift, and tsunami inundation. Because of their geographic location, ports and harbors are especially vulnerable to these hazards. Ports and harbors, however, are important components of many coastal communities, supporting numerous activities critical to the local and regional economy and possibly serving as vital post-event, response-recovery transportation links. A collaborative, multi-year initiative is underway to increase the resiliency of Pacific Northwest ports and harbors to earthquake and tsunami hazards, involving Oregon Sea Grant (OSG), Washington Sea Grant (WSG), the National Oceanic and Atmospheric Administration Coastal Services Center (CSC), and the U.S. Geological Survey Center for Science Policy (CSP). Specific products of this research, planning, and outreach initiative include a regional stakeholder issues and needs assessment, a community-based mitigation planning process, a Geographic Information System (GIS) — based vulnerability assessment methodology, an educational web-site and a regional data archive. This paper summarizes these efforts, including results of two pilot port-harbor community projects, one in Yaquina Bay, Oregon and the other in Sinclair Inlet, Washington. Finally, plans are outlined for outreach to other port and harbor communities in the Pacific Northwest and beyond, using "getting started" workshops and a web-based tutorial.

  7. Stabilization and in situ management of radioactive contaminated sediments of Port Hope harbor

    International Nuclear Information System (INIS)

    Dolinar, G.M.; Killey, R.W.D.; Philipase, K.E.

    1991-01-01

    This paper presents the findings of laboratory and field studies undertaken to assess the feasibility of in situ management of contaminated sediments in Port Hope harbor. The contaminated sediments stem from historic releases from an adjacent radium and uranium refinery, and uranium, arsenic, and radium are the most abundant contaminants. With improved emission controls, currently accumulating sediments have much lower levels of contamination, and the harbor waters currently meet water quality limits for the contaminants of concern. Within a few years, however, the continuing sedimentation will render the harbor unusable. Field tests have confirmed the dredging will result in incomplete removal of the contaminated sediments and that sediment suspension and the release of pores waters during dredging will produce harbor water contaminant concentrations that would require the treatment of large volumes of water. In addition, no remedial work can start until a site for the dredged material can be found. The local community inquired whether in situ burial of the sediments and abandonment of the harbor would provide safe disposal

  8. Seroprevalence of Antibodies against Seal Influenza A(H10N7) Virus in Harbor Seals and Gray Seals from the Netherlands.

    Science.gov (United States)

    Bodewes, Rogier; Rubio García, Ana; Brasseur, Sophie M; Sanchez Conteras, Guillermo J; van de Bildt, Marco W G; Koopmans, Marion P G; Osterhaus, Albert D M E; Kuiken, Thijs

    2015-01-01

    In the spring and summer 2014, an outbreak of seal influenza A(H10N7) virus infection occurred among harbor seals (Phoca vitulina) off the coasts of Sweden and Denmark. This virus subsequently spread to harbor seals off the coasts of Germany and the Netherlands. While thousands of seals were reported dead in Sweden, Denmark and Germany, only a limited number of seals were found dead in the Netherlands. To determine the extent of exposure of seals in the Netherlands to influenza A/H10N7 virus, we measured specific antibody titers in serum samples from live-captured seals and seals admitted for rehabilitation in the Netherlands by use of a hemagglutination inhibition assay and an ELISA. In harbor seals in 2015, antibodies against seal influenza A(H10N7) virus were detected in 41% (32 out of 78) pups, 10% (5 out of 52) weaners, and 58% (7 out of 12) subadults or adults. In gray seals (Halichoerus grypus) in 2015, specific antibodies were not found in the pups (n = 26), but in 26% (5 out of 19) of the older animals. These findings indicate that, despite apparent low mortality, infection with seal influenza A(H10N7) virus was geographically widespread and also occurred in grey seals.

  9. A trans-acting enhancer modulates estrogen-mediated transcription of reporter genes in osteoblasts.

    Science.gov (United States)

    Sasaki-Iwaoka, H; Maruyama, K; Endoh, H; Komori, T; Kato, S; Kawashima, H

    1999-02-01

    The presence of bone-specific estrogen agonists and discovery of the osteoblast-specific transcription factor (TF), Cbfa1, together with the discovery of synergism between a TF Pit-1 and estrogen receptor alpha (ERalpha) on rat prolactin gene, led to investigation of Cbfa1 in the modulation of osteoblast-specific actions of estrogen. Reverse transcribed-polymerase chain reaction demonstrated expression of Cbfa1 in the osteoblastic cell lines, MG63, ROS17/2.8, and MC3T3E1, but not in nonosteoblastic cell lines, MCF7, C3H10T1/2, and HeLa. An ER expression vector and a series of luciferase (Luc) reporter plasmids harboring the Cbfa1 binding site OSE2 (the osteoblast-specific cis element in the osteocalcin promoter) and palindromic estrogen response elements (EREs) were cotransfected into both osteoblastic and nonosteoblastic cells. OSE2 worked as a cis- acting element in osteoblastic cells but not nonosteoblastic cells, whereas EREs were cis- acting in all cell lines. Synergistic transactivation was observed in osteoblastic cells only when both ERE and OSE2 were placed in juxtaposition to the promoter. Forced expression of Cbfa1 in C3H10T1/2 cells also induced synergism. Tamoxifen, a partial agonist/antagonist of estrogen, acted as an osteoblast-specific agonist in cells transfected with a promoter containing ERE and acted synergistically with a promoter containing the ERE-OSE2 enhancer combination. These results support the idea that bone-specific TFs modulate the actions of estrogen in a tissue-specific manner.

  10. Multivariate analysis of heavy metal contaminations in seawater and sediments from a heavily industrialized harbor in Southern Taiwan

    International Nuclear Information System (INIS)

    Lin, Yung-Chang; Chang-Chien, Guo-Ping; Chiang, Pen-Chi; Chen, Wei-Hsiang; Lin, Yuan-Chung

    2013-01-01

    Highlights: • Kaohsiung Harbor is the largest international commercial port in Taiwan. • The metal distributions in the seawater and sediments were investigated. • Many metals exhibited higher levels of enrichment inside the harbor. • Multivariate statistical analysis was used to characterize the metal pollutions. • Two complex arrays of contamination behaviors exist inside and outside the harbor. -- Abstract: Heavy metal pollution, including chromium, zinc, arsenic, cadmium, mercury, copper, lead, and aluminum, in the largest industrial harbor in southern Taiwan was investigated. Increasing metal contamination was observed by monitoring heavy metal concentrations in seawater and sediments and estimating the enrichment factors, particularly those inside the harbor. Compared to other metal-polluted harbors worldwide, the presence of chromium in the sediments was relatively high. Excluding the background contribution, the harbor area was polluted by outflows from river mouths, wastewater discharging pipes, and point sources near industrial activities within the harbor. It is shown by principal component and cluster analyses that metal contamination was affected by a wide range of different and complex contamination mechanisms inside and outside the harbor, suggesting managing the pollution using straightforward strategies, i.e., solutions that only consider a single source or single pathway of metal emissions, is problematic

  11. A neurological comparative study of the harp seal (Pagophilus groenlandicus) and harbor porpoise (Phocoena phocoena) brain

    DEFF Research Database (Denmark)

    Walløe, Solveig; Eriksen, Nina; Dabelsteen, Torben

    2010-01-01

    The cetacean brain is well studied. However, few comparisons have been done with other marine mammals. In this study, we compared the harp seal (Pagophilus groenlandicus) and the harbor porpoise brain (Phocoena phocoena). Stereological methods were applied to compare three areas of interest...... cells, whereas the harp seal have 6.1 × 10(9) neocortical neurons and 17.5 × 10(9) neocortical glial cells. The harbor porpoise have significantly more neurons and glial cells in the auditory cortex than in the visual cortex, whereas the pattern was opposite for the harp seal. These results...... are the first to provide estimates of the number of neurons and glial cells in the neocortex of the harp seal and harbor porpoise brain and offer new data to the comparative field of mammalian brain evolution....

  12. Development of potent ALK inhibitor and its molecular inhibitory mechanism against NSCLC harboring EML4-ALK proteins

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Chung Hyo [Bio & Drug Discovery Division, Korea Research Institute of Chemical Technology, PO Box 107, Daejeon 305-600 (Korea, Republic of); College of Pharmacy, Chungnam National University, Daejeon (Korea, Republic of); Yun, Jeong In [Bio & Drug Discovery Division, Korea Research Institute of Chemical Technology, PO Box 107, Daejeon 305-600 (Korea, Republic of); Lee, Kwangho [Bio & Drug Discovery Division, Korea Research Institute of Chemical Technology, PO Box 107, Daejeon 305-600 (Korea, Republic of); Medicinal & Pharmaceutical Chemistry, Korea University of Science and Technology, Daejeon 305-350 (Korea, Republic of); Lee, Chong Ock; Lee, Heung Kyoung [Bio & Drug Discovery Division, Korea Research Institute of Chemical Technology, PO Box 107, Daejeon 305-600 (Korea, Republic of); Yun, Chang-Soo; Hwang, Jong Yeon; Cho, Sung Yun; Jung, Heejung; Kim, Pilho [Bio & Drug Discovery Division, Korea Research Institute of Chemical Technology, PO Box 107, Daejeon 305-600 (Korea, Republic of); Medicinal & Pharmaceutical Chemistry, Korea University of Science and Technology, Daejeon 305-350 (Korea, Republic of); Ha, Jae Du; Jeon, Jeong Hee; Choi, Sang Un [Bio & Drug Discovery Division, Korea Research Institute of Chemical Technology, PO Box 107, Daejeon 305-600 (Korea, Republic of); Jeong, Hye Gwang [College of Pharmacy, Chungnam National University, Daejeon (Korea, Republic of); Kim, Hyoung Rae, E-mail: hyungrk@krict.re.kr [Bio & Drug Discovery Division, Korea Research Institute of Chemical Technology, PO Box 107, Daejeon 305-600 (Korea, Republic of); Park, Chi Hoon, E-mail: chpark@krict.re.kr [Bio & Drug Discovery Division, Korea Research Institute of Chemical Technology, PO Box 107, Daejeon 305-600 (Korea, Republic of); Medicinal & Pharmaceutical Chemistry, Korea University of Science and Technology, Daejeon 305-350 (Korea, Republic of)

    2015-08-28

    Here, we show the newly synthesized and potent ALK inhibitor having similar scaffold to KRCA-0008, which was reported previously, and its molecular mechanism against cancer cells harboring EML4-ALK fusion protein. Through ALK wild type enzyme assay, we selected two compounds, KRCA-0080 and KRCA-0087, which have trifluoromethyl instead of chloride in R2 position. We characterized these newly synthesized compounds by in vitro and in vivo assays. Enzyme assay shows that KRCA-0080 is more potent against various ALK mutants, including L1196M, G1202R, T1151-L1152insT, and C1156Y, which are seen in crizotinib-resistant patients, than KRCA-0008 is. Cell based assays demonstrate our compounds downregulate the cellular signaling, such as Akt and Erk, by suppressing ALK activity to inhibit the proliferation of the cells harboring EML4-ALK. Interestingly, our compounds induced strong G1/S arrest in H3122 cells leading to the apoptosis, which is proved by PARP-1 cleavage. In vivo H3122 xenograft assay, we found that KRCA-0080 shows significant reduction in tumor size compared to crizotinib and KRCA-0008 by 15–20%. Conclusively, we report a potent ALK inhibitor which shows significant in vivo efficacy as well as excellent inhibitory activity against various ALK mutants. - Highlights: • We synthesized KRCA-0008 derivatives having trifluoromethyl instead of chloride. • KRCA-0080 shows superior activity against several ALK mutants to KRCA-0008. • Cellular assays show our ALK inhibitors suppress only EML4-ALK positive cells. • Our ALK inhibitors induce G1/S arrest to lead apoptosis in H3122 cells. • KRCA-0080 has superior in vivo efficacy to crizotinib and KRCA-0008 by 15–20%.

  13. Safe Harbor Legislation for Juvenile Victims of Sex Trafficking: A Myopic View of Improvements in Practice

    Directory of Open Access Journals (Sweden)

    Kimberly Mehlman-Orozco

    2015-02-01

    Full Text Available Current social and political realties have focused attention on human trafficking in the United States. Although new mechanisms for criminalizing offenders and protecting victims are increasingly funded and implemented across the country, empirical exploration into the efficacy of these interventions is lacking. This article uses yearly count data on juvenile prostitution arrests aggregated at the state level to explore the criminalization of commercial sexually exploited children post safe harbor policy implementation. Preliminary data from four states suggests that the passage of safe harbor laws may not reduce the number of juveniles arrested for prostitution crimes. Implications for future research are discussed.

  14. Ultra-High Foraging Rates of Harbor Porpoises Make Them Vulnerable to Anthropogenic Disturbance.

    Science.gov (United States)

    Wisniewska, Danuta Maria; Johnson, Mark; Teilmann, Jonas; Rojano-Doñate, Laia; Shearer, Jeanne; Sveegaard, Signe; Miller, Lee A; Siebert, Ursula; Madsen, Peter Teglberg

    2016-06-06

    The question of how individuals acquire and allocate resources to maximize fitness is central in evolutionary ecology. Basic information on prey selection, search effort, and capture rates are critical for understanding a predator's role in its ecosystem and for predicting its response to natural and anthropogenic disturbance. Yet, for most marine species, foraging interactions cannot be observed directly. The high costs of thermoregulation in water require that small marine mammals have elevated energy intakes compared to similar-sized terrestrial mammals [1]. The combination of high food requirements and their position at the apex of most marine food webs may make small marine mammals particularly vulnerable to changes within the ecosystem [2-4], but the lack of detailed information about their foraging behavior often precludes an informed conservation effort. Here, we use high-resolution movement and prey echo recording tags on five wild harbor porpoises to examine foraging interactions in one of the most metabolically challenged cetacean species. We report that porpoises forage nearly continuously day and night, attempting to capture up to 550 small (3-10 cm) fish prey per hour with a remarkable prey capture success rate of >90%. Porpoises therefore target fish that are smaller than those of commercial interest, but must forage almost continually to meet their metabolic demands with such small prey, leaving little margin for compensation. Thus, for these "aquatic shrews," even a moderate level of anthropogenic disturbance in the busy shallow waters they share with humans may have severe fitness consequences at individual and population levels. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Fanconi anemia core complex gene promoters harbor conserved transcription regulatory elements.

    Science.gov (United States)

    Meier, Daniel; Schindler, Detlev

    2011-01-01

    The Fanconi anemia (FA) gene family is a recent addition to the complex network of proteins that respond to and repair certain types of DNA damage in the human genome. Since little is known about the regulation of this novel group of genes at the DNA level, we characterized the promoters of the eight genes (FANCA, B, C, E, F, G, L and M) that compose the FA core complex. The promoters of these genes show the characteristic attributes of housekeeping genes, such as a high GC content and CpG islands, a lack of TATA boxes and a low conservation. The promoters functioned in a monodirectional way and were, in their most active regions, comparable in strength to the SV40 promoter in our reporter plasmids. They were also marked by a distinctive transcriptional start site (TSS). In the 5' region of each promoter, we identified a region that was able to negatively regulate the promoter activity in HeLa and HEK 293 cells in isolation. The central and 3' regions of the promoter sequences harbor binding sites for several common and rare transcription factors, including STAT, SMAD, E2F, AP1 and YY1, which indicates that there may be cross-connections to several established regulatory pathways. Electrophoretic mobility shift assays and siRNA experiments confirmed the shared regulatory responses between the prominent members of the TGF-β and JAK/STAT pathways and members of the FA core complex. Although the promoters are not well conserved, they share region and sequence specific regulatory motifs and transcription factor binding sites (TBFs), and we identified a bi-partite nature to these promoters. These results support a hypothesis based on the co-evolution of the FA core complex genes that was expanded to include their promoters.

  16. Fanconi anemia core complex gene promoters harbor conserved transcription regulatory elements.

    Directory of Open Access Journals (Sweden)

    Daniel Meier

    Full Text Available The Fanconi anemia (FA gene family is a recent addition to the complex network of proteins that respond to and repair certain types of DNA damage in the human genome. Since little is known about the regulation of this novel group of genes at the DNA level, we characterized the promoters of the eight genes (FANCA, B, C, E, F, G, L and M that compose the FA core complex. The promoters of these genes show the characteristic attributes of housekeeping genes, such as a high GC content and CpG islands, a lack of TATA boxes and a low conservation. The promoters functioned in a monodirectional way and were, in their most active regions, comparable in strength to the SV40 promoter in our reporter plasmids. They were also marked by a distinctive transcriptional start site (TSS. In the 5' region of each promoter, we identified a region that was able to negatively regulate the promoter activity in HeLa and HEK 293 cells in isolation. The central and 3' regions of the promoter sequences harbor binding sites for several common and rare transcription factors, including STAT, SMAD, E2F, AP1 and YY1, which indicates that there may be cross-connections to several established regulatory pathways. Electrophoretic mobility shift assays and siRNA experiments confirmed the shared regulatory responses between the prominent members of the TGF-β and JAK/STAT pathways and members of the FA core complex. Although the promoters are not well conserved, they share region and sequence specific regulatory motifs and transcription factor binding sites (TBFs, and we identified a bi-partite nature to these promoters. These results support a hypothesis based on the co-evolution of the FA core complex genes that was expanded to include their promoters.

  17. Harbor Seal (Phoca vitulina) Reproductive Advertisement Behavior and the Effects of Vessel Noise

    Science.gov (United States)

    Matthews, Leanna P.

    Harbor seals (Phoca vitulina) are a widely distributed pinniped species that mate underwater. Similar to other aquatically mating pinnipeds, male harbor seals produce vocalizations during the breeding season that function in male-male interactions and possibly as an attractant for females. I investigated multiple aspects of these reproductive advertisement displays in a population of harbor seals in Glacier Bay National Park and Preserve, Alaska. First, I looked at vocal production as a function of environmental variables, including season, daylight, and tidal state. Vocalizations were highly seasonal and detection of these vocalizations peaked in June and July, which correspond with the estimated time of breeding. Vocalizations also varied with light, with the lowest probability of detection during the day and the highest probability of detection at night. The high probability of detection corresponded to when females are known to forage. These results are similar to the vocal behavior of previously studied populations. However, unlike previously studied populations, the detection of harbor seal breeding vocalizations did not vary with tidal state. This is likely due to the location of the hydrophone, as it was not near the haul out and depth was therefore not significantly influenced by changes in tidal height. I also investigated the source levels and call parameters of vocalizations, as well as call rate and territoriality. The average source level of harbor seal breeding vocalizations was 144 dB re 1 ?Pa at 1 m and measurements ranged from 129 to 149 dB re 1 ?Pa. Analysis of call parameters indicated that vocalizations of harbor seals in Glacier Bay were similar in duration to other populations, but were much lower in frequency. During the breeding season, there were two discrete calling areas that likely represent two individual males; the average call rate in these display areas was approximately 1 call per minute. The harbor seal breeding season also

  18. Operation and Maintence, Vermilion Harbor, Erie County, Ohio.

    Science.gov (United States)

    1976-03-01

    Aphanozomenon flos- aguae . The Ohio EPA reports that on a yearly average, Cyclotella, Stephanodiscus and pennate diatoms are predominant, particularly during the...for both recre- ational and potable water uses (71). For recreational uses (swimming, boating, skiing, etc.), the number of total coliforms per 100 ml

  19. Serratia marcescens harboring SME-4 in Brazil: A silent threat.

    Science.gov (United States)

    Cayô, Rodrigo; Leme, Rodrigo Cuiabano Paes; Streling, Ana Paula; Matos, Adriana Pereira; Nodari, Carolina Silva; Chaves, Jessica Reis Esteves; Brandão, Jorge Luiz Ferreira; de Almeida, Maíra Fernandes; Carrareto, Valério; de Castro Pereira, Marco Aurélio; de Almeida, Jean Pierre Aquino; Ferreira, Demian Candido; Gales, Ana Cristina

    2017-04-01

    The intrinsic polymyxin resistance displayed by Serratia marcescens makes the acquisition of carbapenemase encoding genes a worrisome event. This study report a SME-4-producing S. marcescens isolate causing septic shock in Brazil. The insertion of novel resistance determinants and their consequent spread in our territory is noteworthy. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. 26 CFR 1.475(a)-4 - Valuation safe harbor.

    Science.gov (United States)

    2010-04-01

    ... being marked to market under section 475 is the primary financial statement. (5) Consolidated groups. If... values of positions reported on certain financial statements as the fair market values of those positions... if the financial statement employing that method has certain indicia of reliability, then the values...

  1. Shoaling Analysis at Brazos Island, Harbor Inlet, Texas

    Science.gov (United States)

    2015-10-01

    and, consequently, is not a high priority for maintenance dredging funding. However, any impacts to inbound vessels ensure a major negative economic...Brownsville Ship Channel. Corpus Christi, TX. Report prepared for Port of Brownsville, TX. Martin Associates. 2008. Market assessment of the Port of

  2. Colistin- and Carbapenem-Resistant Escherichia coli Harboring mcr-1 and blaNDM-5, Causing a Complicated Urinary Tract Infection in a Patient from the United States

    Directory of Open Access Journals (Sweden)

    José R. Mediavilla

    2016-08-01

    Full Text Available Colistin is increasingly used as an antibiotic of last resort for the treatment of carbapenem-resistant Gram-negative infections. The plasmid-borne colistin resistance gene mcr-1 was initially identified in animal and clinical samples from China and subsequently reported worldwide, including in the United States. Of particular concern is the spread of mcr-1 into carbapenem-resistant bacteria, thereby creating strains that approach pan-resistance. While several reports of mcr-1 have involved carbapenem-resistant strains, no such isolates have been described in the United States. Here, we report the isolation and identification of an Escherichia coli strain harboring both mcr-1 and carbapenemase gene blaNDM-5 from a urine sample in a patient without recent travel outside the United States. The isolate exhibited resistance to both colistin and carbapenems, but was susceptible to amikacin, aztreonam, gentamicin, nitrofurantoin, tigecycline, and trimethoprim-sulfamethoxazole. The mcr-1- and blaNDM-5-harboring plasmids were completely sequenced and shown to be highly similar to plasmids previously reported from China. The strain in this report was first isolated in August 2014, highlighting an earlier presence of mcr-1 within the United States than previously recognized.

  3. Reporter gene expression in fish following cutaneous infection with pantropic retroviral vectors.

    Science.gov (United States)

    Paul, T A; Burns, J C; Shike, H; Getchell, R; Bowser, P R; Whitlock, K E; Casey, J W

    2001-06-01

    A central issue in gene delivery systems is choosing promoters that will direct defined and sustainable levels of gene expression. Pantropic retroviral vectors provide a means to insert genes into either somatic or germline cells. In this study, we focused on somatic cell infection by evaluating the activity of 3 promoters inserted by vectors into fish cell lines and fish skin using pantropic retroviruses. In bluegill and zebrafish cell lines, the highest levels of luciferase expression were observed from the 5' murine leukemia virus long terminal repeat of the retroviral vector. The Rous sarcoma virus long terminal repeat and cytomegalovirus early promoter, as internal promoters, generated lower levels of luciferase. Luciferase reporter vectors infected zebrafish skin, as measured by the presence of viral DNA, and expressed luciferase. We infected developing walleye dermal sarcomas with retroviral vectors to provide an environment with enhanced cell proliferation, a condition necessary for integration of the provirus into the host genome. We demonstrated a 4-fold to 7-fold increase in luciferase gene expression in tumor tissue over infections in normal walleye skin.

  4. Identification and characterization of the Luc2-type luciferase in the Japanese firefly, Luciola parvula, involved in a dim luminescence in immobile stages.

    Science.gov (United States)

    Bessho-Uehara, Manabu; Oba, Yuichi

    2017-09-01

    Nocturnal Japanese fireflies, Luciola parvula, emit from their lanterns a yellow light, one of the most red-shifted colors found among fireflies. Previously, we isolated and characterized two different types of luciferase gene, Luc1 and Luc2, from the fireflies Luciola cruciata and Luciola lateralis; Luc1 is responsible for the green-yellow luminescence of larval and adult lanterns, whereas Luc2 is responsible for the dim greenish glow of eggs and pupal bodies. The biological role of firefly lanterns in adults is related to sexual communication, but why the eggs and pupae glow remains uncertain. In this study, we isolated the gene Luc2 from L. parvula, and compared its expression profiles and enzymatic characteristics with those of Luc1. A semi-quantitative reverse transcription polymerase chain reaction showed that Luc1 was predominantly expressed in larvae, prepupae, pupae and adults, whereas Luc2 was expressed in eggs, prepupae, pupae and adult females. Enzymatic analyses showed that the luminescent color of Luc1 matches the visual sensitivity of L. parvula eyes, whereas that of Luc2 is very different from it. These results suggest that the biological role of Luc2 expressed in immobile stages is not intraspecific communication. Copyright © 2017 John Wiley & Sons, Ltd.

  5. [Establishment of an iRFP and luciferase dual-color fluorescence-traced hepatocellular carcinoma transplantation model in nude mice].

    Science.gov (United States)

    Li, Hongjun; Yang, Tianhua; Huang, Yanping; Liu, Mingzhu; Qin, Zhongqiang; Chu, Fei; Li, Zhenghong; Li, Yonghai

    2017-11-01

    Objective To establish a hepatocellular carcinoma xenograft model in nude mice which could stably express gene and be monitored dynamically. Methods We first constructed the lentiviral particles containing luciferase (Luc) and near-infrared fluorescent protein (iRFP) and puromycin resistance gene, and then transduced them into the HepG2 hepatoma cells. The cell line stably expressing Luc and iRFP genes were screened and inoculated into nude mice to establish xenograft tumor model. Tumor growth was monitored using in vivo imaging system. HE staining and immunohistochemistry were used to evaluate the pathological features and tumorigenic ability. Results HepG2 cells stably expressing iRFP and Luc were obtained; with the engineered cell line, xenograft model was successfully established with the features of proper tumor developing time and high rate of tumor formation as well as typical pathological features as showed by HE staining and immunohistochemistry. Conclusion Hepatocellular carcinoma model in nude mice with the features of stable gene expression and dynamical monitoring has been established successfully with the HepG2-iRFP-Luc cell line.

  6. A dual function fusion protein of Herpes simplex virus type 1 thymidine kinase and firefly luciferase for noninvasive in vivo imaging of gene therapy in malignant glioma.

    Science.gov (United States)

    Söling, Ariane; Theiss, Christian; Jungmichel, Stephanie; Rainov, Nikolai G

    2004-08-04

    BACKGROUND: Suicide gene therapy employing the prodrug activating system Herpes simplex virus type 1 thymidine kinase (HSV-TK)/ ganciclovir (GCV) has proven to be effective in killing experimental brain tumors. In contrast, glioma patients treated with HSV-TK/ GCV did not show significant treatment benefit, most likely due to insufficient transgene delivery to tumor cells. Therefore, this study aimed at developing a strategy for real-time noninvasive in vivo monitoring of the activity of a therapeutic gene in brain tumor cells. METHODS: The HSV-TK gene was fused to the firefly luciferase (Luc) gene and the fusion construct HSV-TK-Luc was expressed in U87MG human malignant glioma cells. Nude mice with subcutaneous gliomas stably expressing HSV-TK-Luc were subjected to GCV treatment and tumor response to therapy was monitored in vivo by serial bioluminescence imaging. Bioluminescent signals over time were compared with tumor volumes determined by caliper. RESULTS: Transient and stable expression of the HSV-TK-Luc fusion protein in U87MG glioma cells demonstrated close correlation of both enzyme activities. Serial optical imaging of tumor bearing mice detected in all cases GCV induced death of tumor cells expressing the fusion protein and proved that bioluminescence can be reliably used for repetitive and noninvasive quantification of HSV-TK/ GCV mediated cell kill in vivo. CONCLUSION: This approach may represent a valuable tool for the in vivo evaluation of gene therapy strategies for treatment of malignant disease.

  7. Preliminary Feasibility Report Stage II, Lorain Small-Boat Harbor, Lorain, Ohio. Revision.

    Science.gov (United States)

    1982-03-01

    frequently trailered, increasing numbers of auxillary powered sailboats and smaller, non- auxillary powered sailboats are creating excess demand for faci...in/outboards, 8 percent inboards, 16 percent sailboats, 8 percent auxillary sailboats, and 40 percent cruisers. The projected fleet mix by size is

  8. Burlington Northern Taconite Transshipment Facility, Duluth-Superior Harbor, Superior Wisconsin. Environmental Assessment Report.

    Science.gov (United States)

    1975-03-01

    the Federal Government declared the Duluth- Superior area to be economically depressed . The reason given was the "consistant and chronic unemployment...include dogwood, sumac, arrowwood, blueberry, highbush cranberry , elderberry, wild grape, buttonbrush, snowberry and partridgeberry. Aquatic and...water for the proposed greenbelt areas and as dust sup- pression spray. 10.003 The depressed economy of the Duluth-Superior area would benefit by the

  9. Vermilion Harbor, Ohio. Detailed Project Report on Section 111, Shore Erosion Study. Stage 3 Documentation

    Science.gov (United States)

    1982-09-01

    be the implementation of pollution abatement measures throughout the Vermilion region to reduce the addition of toxic and nutritive constituents to...Breakwater Impact Study - Study of the Impact of the Offshore Breakwater on: Municipal Water Supply; Swiming Area and Beaches; 3 OF 6 ". .*., Ice Jam Flooding

  10. Biological Survey, Buffalo River and Outer Harbor of Buffalo, New York. Volume II. Data Report.

    Science.gov (United States)

    1982-06-01

    Arctium mimus Schk. (burdock) 3 3 Leonurus cardiaca L. 3 (motherwo-3 Oenothera biennia L. 3 3 (evening pri-rose) Galium AM ine L. 2 (bedstaw) Asclapias...Wild carrot Bromus japonicus Thumb. Japanese brome , Oenothera biennis L. Evening primrose Clematis virginiana Virgin’ s bower Bidens tripartita Beggar

  11. 76 FR 27970 - Safety Zone; Cape Charles Fireworks, Cape Charles Harbor, Cape Charles, VA.

    Science.gov (United States)

    2011-05-13

    ... Charles will sponsor a fireworks display on the shoreline of the navigable waters of Cape Charles City...[deg]01'30'' W (NAD 1983). This safety zone will be established in the vicinity of Cape Charles, VA...-AA00 Safety Zone; Cape Charles Fireworks, Cape Charles Harbor, Cape Charles, VA. AGENCY: Coast Guard...

  12. 75 FR 38019 - Safety Zone; Fixed Mooring Balls, South of Barbers Pt. Harbor Channel, Oahu, HI

    Science.gov (United States)

    2010-07-01

    ... DEPARTMENT OF HOMELAND SECURITY Coast Guard 33 CFR Part 165 [Docket No. USCG-2010-0457] RIN 1625-AA00 Safety Zone; Fixed Mooring Balls, South of Barbers Pt. Harbor Channel, Oahu, HI AGENCY: Coast Guard, DHS. ACTION: Temporary final rule. SUMMARY: Due to the placement of six fixed mooring balls in an...

  13. 75 FR 56130 - Pearl Harbor National Wildlife Refuge, Honolulu County, HI; Comprehensive Conservation Plan and...

    Science.gov (United States)

    2010-09-15

    ... DEPARTMENT OF THE INTERIOR Fish and Wildlife Service [FWS-R1-R-2010-N160; 1265-0000-10137-S3] Pearl Harbor National Wildlife Refuge, Honolulu County, HI; Comprehensive Conservation Plan and... Kamehameha Highway, Room 2C, Hale`iwa, HI 96712. Alternatively, you may fax comments to the refuge at (808...

  14. Side-scan imagery of Apra Harbor, Guam U.S. Territory

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Gridded side-scan imagery collected in Apra Harbor, Guam U.S. Territory, USA. The GeoTIFF and Arc ASCII file includes data that were collected using a Klein 3000...

  15. Evaluation of Proposed Dredged Material Disposal Alternatives for New York/New Jersey Harbor. Phase 1

    Science.gov (United States)

    1988-08-01

    been developed ( Westerdahl and Skogerboe 1981) and is being used x..-’. to predict surface runoff water quality from dredged material as part of the...Islands in New York Harbor," Miscellaneous Paper D-88-3, US Army Engineer Waterways Experiment Station, Vicksburg, MS. Westerdahl , H. E., and Skogerboe

  16. Application of heterogeneous multiple camera system with panoramic capabilities in a harbor environment

    NARCIS (Netherlands)

    Schwering, P.B.W.; Lensen, H.A.; Broek, S.P. van den; Hollander, R.J.M. den; Mark, W. van der; Bouma, H.; Kemp, R.A.W.

    2009-01-01

    In a harbor environment threats like explosives-packed rubber boats, mine-carrying swimmers and divers must be detected in an early stage. This paper describes the integration and use of a heterogeneous multiple camera system with panoramic observation capabilities for detecting these small vessels

  17. Cold Spring Harbor symposia on quantitative biology: Volume 51, Molecular biology of /ital Homo sapiens/

    International Nuclear Information System (INIS)

    1986-01-01

    This volume is the second part of a collection of papers submitted by the participants to the 1986 Cold Spring Harbor Symposium on Quantitative Biology entitled Molecular Biology of /ital Homo sapiens/. The 49 papers included in this volume are grouped by subject into receptors, human cancer genes, and gene therapy. (DT)

  18. Mitigation of Shore Damage Attributed to the Federal Navigation Structures at Port Sanilac Harbor, Michigan.

    Science.gov (United States)

    1975-09-01

    produces waves with nealioible wace heights (i.e., 0.5 feet); (b) all waves were deep water waves with no correction for shallow water; and, (c) the...at that harbor. This would re- sult in a loss to the local economy . Ini addition, littoral drift would resume southward thus affecting the littoral

  19. Factors Contributing to Corrosion of Steel Pilings in Duluth-Superior Harbor

    Science.gov (United States)

    2009-11-01

    1226 Office of Counsel,Code 1008.3 ADOR/Director NCST E. R. Franchi , 7000 Public Affairs (Unclassified/ Unlimited Only), Code 703o 4...Great Lakes. Accelerated corrosion of CS pilings in estua- rine and marine harbors is a global phenomenon.9 The term "accelerated low water corrosion

  20. 78 FR 18481 - Drawbridge Operation Regulations; Inner Harbor Navigation Canal, LA

    Science.gov (United States)

    2013-03-27

    ... regulation governing the operation of the US 90 (Danzinger) vertical lift span drawbridge across the Inner... Lift Span Bridge across the Inner Harbor Navigation Canal, mile 3.10 at New Orleans, Orleans Parish, Louisiana. The bridge has a vertical clearance of 50 feet above mean high water, elevation 5.0 feet Mean Sea...

  1. 77 FR 42640 - Safety Zone; Can-Am Festival Fireworks, Black River Bay, Sackets Harbor, NY

    Science.gov (United States)

    2012-07-20

    ...-AA00 Safety Zone; Can-Am Festival Fireworks, Black River Bay, Sackets Harbor, NY AGENCY: Coast Guard... Black River Bay during the Can-Am Festival Fireworks display. This temporary safety zone is necessary to... Can-Am Festival Fireworks. This zone will be effective and enforced from 9:15 p.m. until 10:45 p.m. on...

  2. 33 CFR 110.190 - Tortugas Harbor, in vicinity of Garden Key, Dry Tortugas, Fla.

    Science.gov (United States)

    2010-07-01

    ... Garden Key, Dry Tortugas, Fla. 110.190 Section 110.190 Navigation and Navigable Waters COAST GUARD..., in vicinity of Garden Key, Dry Tortugas, Fla. (a) The anchorage grounds. All of Bird Key Harbor, southwest of Garden Key, bounded by the surrounding reefs and shoals and, on the northeast, by a line...

  3. 78 FR 62439 - Drawbridge Operation Regulation; Inner Harbor Navigational Canal, New Orleans, LA

    Science.gov (United States)

    2013-10-22

    ... Operation Regulation; Inner Harbor Navigational Canal, New Orleans, LA AGENCY: Coast Guard, DHS. ACTION..../Seabrook) bridge across the IHNC, mile 4.6, both at New Orleans, LA. This deviation will test changes to... Intra-Costal Waterway (GIWW). The US 90 (Danzinger) Bridge across the IHNC, mile 3.1, at New Orleans...

  4. 75 FR 8486 - Drawbridge Operation Regulation; Inner Harbor Navigational Canal, New Orleans, LA

    Science.gov (United States)

    2010-02-25

    ... Operation Regulation; Inner Harbor Navigational Canal, New Orleans, LA AGENCY: Coast Guard, DHS. ACTION... EHL), at New Orleans, LA. The deviation is necessary to replace the counterweight wire ropes on the... Building Ground Floor, Room W12-140, 1200 New Jersey Avenue, SE., Washington, DC 20590, between 9 a.m. and...

  5. 78 FR 63136 - Drawbridge Operation Regulation; Inner Harbor Navigational Canal, New Orleans, LA

    Science.gov (United States)

    2013-10-23

    ...-AA09 Drawbridge Operation Regulation; Inner Harbor Navigational Canal, New Orleans, LA AGENCY: Coast... 4.6, both at New Orleans, LA. This proposed change would allow for the safe navigation of vessels... Department of Transportation and Development (LDOTD) (representing the New Orleans Levee District which is...

  6. 3 CFR 8463 - Proclamation 8463 of December 4, 2009. National Pearl Harbor Remembrance Day, 2009

    Science.gov (United States)

    2010-01-01

    ... who selflessly served our Nation at home and abroad during World War II. On a tranquil Sunday morning, as war raged around the globe, the attack on Pearl Harbor effectively ended American isolation... have hereunto set my hand this fourth day of December, in the year of our Lord two thousand nine, and...

  7. 76 FR 4574 - Drawbridge Operation Regulations; Oakland Inner Harbor Tidal Canal, Oakland/Alameda, CA, Schedule...

    Science.gov (United States)

    2011-01-26

    ... Boating Federation, Hanson Aggregates, Power Engineering, Harbor Bay Maritime, Dutra Group, Oakland Yacht... Association, Aeolian Yacht Club, Briar Rose Yacht Charters, Baytech Marine Service, Heinold's First and Last... Outboard Motor Shop, Waterfront Hotel-Miss Pearl's Restaurant, Encinal Yacht Club, Marina Village Inn...

  8. 78 FR 24069 - Safety Zone; 12th Annual Saltwater Classic; Port Canaveral Harbor; Port Canaveral, FL

    Science.gov (United States)

    2013-04-24

    ... under Executive Order 13045, Protection of Children from Environmental Health Risks and Safety Risks. This rule is not an economically significant rule and does not create an environmental risk to health...-AA00 Safety Zone; 12th Annual Saltwater Classic; Port Canaveral Harbor; Port Canaveral, FL AGENCY...

  9. 78 FR 27304 - Safety Zone; Melrose Pyrotechnics Fireworks Display; Chicago Harbor, Chicago, IL

    Science.gov (United States)

    2013-05-10

    ... environmental risk to health or risk to safety that may disproportionately affect children. 11. Indian Tribal... Pier in Chicago Harbor, Chicago, IL. The Captain of the Port, Lake Michigan, has determined that these fireworks displays will pose a significant risk to public safety and property. Such hazards include falling...

  10. 78 FR 52753 - Taking of Marine Mammals Incidental to Commercial Fishing Operations; Harbor Porpoise Take...

    Science.gov (United States)

    2013-08-26

    ... strategy enacted in 2010 based on deliberations by the Harbor Porpoise Take Reduction Team. This action is... information (e.g., name, address, etc.), confidential business information, or otherwise sensitive information... Protection Act (MMPA) to reduce the level of serious injury and mortality of the Gulf of Maine/Bay of Fundy...

  11. Observations and modeling of tsunami-induced currents in ports and harbors

    Science.gov (United States)

    Lynett, Patrick J.; Borrero, Jose C.; Weiss, Robert; Son, Sangyoung; Greer, Dougal; Renteria, Willington

    2012-04-01

    Tsunamis, or "harbor waves" in Japanese, are so-named due to common observations of enhanced wave heights, currents and damage in harbors and ports. However, dynamic currents induced by these waves, while regularly observed and known to cause significant damage, are poorly understood. Observations and modeling of the currents induced by the 2011 Tohoku and 2004 Indian Ocean tsunamis allows us to show that the strongest flows in harbor basins are governed by horizontally sheared and rotational shallow features, such as jets and large eddies. When examining currents in harbors, this conclusion will generally require a simulation approach that both includes the relevant physical processes in the governing equations and uses a numerical scheme that does not artificially damp these features. Without proper representation of the physics associated with these phenomena, predictive models may provide drag force estimates that are an order of magnitude or more in error. The immediate implementation of this type of analysis into tsunami hazard studies can mean the difference between an unaffected port and one in which 300 m long container vessels are detached from their moorings and drift chaotically.

  12. Surveillance for zoonotic and selected pathogens in harbor seals Phoca vitulina from central California

    Science.gov (United States)

    Greig, Denise J.; Ip, Hon S.; Gulland, Frances M. D.; Miller, Woutrina A.; Conrad, Patricia A.; Field, Cara L.; Fleetwood, Michelle; Harvey, James T.; Jang, Spencer; Packham, Andrea; Wheeler, Elizabeth; Hall, Ailsa J.

    2014-01-01

    The infection status of harbor seals Phoca vitulina in central California, USA, was evaluated through broad surveillance for pathogens in stranded and wild-caught animals from 2001 to 2008, with most samples collected in 2007 and 2008. Stranded animals from Mendocino County to San Luis Obispo County were sampled at a rehabilitation facility: The Marine Mammal Center (TMMC, n = 175); wild-caught animals were sampled at 2 locations: San Francisco Bay (SF, n = 78) and Tomales Bay (TB, n = 97), that differed in degree of urbanization. Low prevalences of Salmonella, Campylobacter, Giardia, and Cryptosporidium were detected in the feces of stranded and wild-caught seals. Clostridium perfringens and Escherichia coli were more prevalent in the feces of stranded (58% [78 out of 135] and 76% [102 out of 135]) than wild-caught (42% [45 out of 106] and 66% [68 out of 106]) seals, whereas Vibrio spp. were 16 times more likely to be cultured from the feces of seals from SF than TB or TMMC (p neurona, and type A influenza was only detected in the wild-caught harbor seals (post-weaning age classes), whereas antibody titers to Leptospira spp. were detected in stranded and wild-caught seals. No stranded (n = 109) or wild-caught (n = 217) harbor seals had antibodies to phocine distemper virus, although a single low titer to canine distemper virus was detected. These results highlight the role of harbor seals as sentinel species for zoonotic and terrestrial pathogens in the marine environment.

  13. Influenza A (H10N7) Virus Causes Respiratory Tract Disease in Harbor Seals and Ferrets

    NARCIS (Netherlands)

    van den Brand, Judith M A; Wohlsein, Peter; Herfst, Sander; Bodewes, Rogier; Pfankuche, Vanessa M; van de Bildt, Marco W G; Seehusen, Frauke; Puff, Christina; Richard, Mathilde; Siebert, Ursula; Lehnert, Kristina; Bestebroer, Theo; Lexmond, Pascal; Fouchier, Ron A M; Prenger-Berninghoff, Ellen; Herbst, Werner; Koopmans, Marion; Osterhaus, Albert D M E; Kuiken, Thijs; Baumgärtner, Wolfgang

    2016-01-01

    Avian influenza viruses sporadically cross the species barrier to mammals, including humans, in which they may cause epidemic disease. Recently such an epidemic occurred due to the emergence of avian influenza virus of the subtype H10N7 (Seal/H10N7) in harbor seals (Phoca vitulina). This epidemic

  14. Sedimentation and chronology of heavy metal pollution in Oslo harbor, Norway

    DEFF Research Database (Denmark)

    Lepland, Aivo; Andersen, Thorbjørn Joest; Lepland, Aave

    2010-01-01

    Stratigraphic profiles of Cu, Cd and Hg in ten sediment cores from the Oslo harbor, Norway, combined with results of radiometric dating demonstrate that pollution by these metals peaked between 1940 and 1970. Dating results indicate that Hg discharges peaked between 1940 and 1950, Cd reached...

  15. Seroprevalence of Brucella antibodies in harbor seals in Alaska, USA, with age, regional, and reproductive comparisons.

    Science.gov (United States)

    Hoover-Miller, A; Dunn, J L; Field, C L; Blundell, G; Atkinson, S

    2017-09-20

    Populations of harbor seal Phoca vitulina in the Gulf of Alaska have dramatically declined during the past 4 decades. Numbers of seals in Glacier Bay, in southeast Alaska, USA, have also declined despite extensive protection. Causes of the declines and slow recovery are poorly understood. Brucellosis is a zoonotic disease that adversely affects reproduction in many domestic species. We measured the seroprevalence of Brucella antibodies in 554 harbor seals in 3 Alaska locations: Prince William Sound (PWS), Glacier Bay (GB), and Tracy Arm Fords Terror (TAFT) Wilderness Area. Objectives included testing for regional, sex, age, and female reproductive state differences in Brucella antibody seroprevalence, persistence in titers in recaptured seals, and differences in titers between mother seals and their pups. Overall, 52% of adults (AD), 53% of subadults (SA), 77% of yearlings (YRL), and 26% of Brucella. Results show higher seroprevalence (64%) for AD and SA seals in the depressed and declining populations in PWS and GB than in TAFT (29%). Lactating females were less likely to be seropositive than other AD females, including pregnant females. Further research is needed to seek evidence of Brucella infection in Alaskan harbor seals, identify effects on neonatal viability, and assess zoonotic implications for Alaska Natives who rely on harbor seals for food.

  16. 33 CFR 117.213 - New Haven Harbor, Quinnipiac and Mill Rivers.

    Science.gov (United States)

    2010-07-01

    ... Mill Rivers. 117.213 Section 117.213 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF... Haven Harbor, Quinnipiac and Mill Rivers. The draws of the Tomlinson bridge, mile 0.0, the Ferry Street... bridge, mile 0.4 across Mill River, shall operate as follows: (a) The draws shall open on signal; except...

  17. 75 FR 1738 - Drawbridge Operation Regulation; New Haven Harbor, Quinnipiac and Mill Rivers, CT

    Science.gov (United States)

    2010-01-13

    ...-AA09 Drawbridge Operation Regulation; New Haven Harbor, Quinnipiac and Mill Rivers, CT AGENCY: Coast... regulation governing the operation of three bridges across the Quinnipiac and Mill Rivers at New Haven... and 15 feet at mean low water. The Chapel Street Bridge at mile 0.4, across the Mill River has a...

  18. 77 FR 75917 - Drawbridge Operation Regulation; New Haven Harbor, Quinnipiac and Mill Rivers, CT

    Science.gov (United States)

    2012-12-26

    ...-AA09 Drawbridge Operation Regulation; New Haven Harbor, Quinnipiac and Mill Rivers, CT AGENCY: Coast..., mile 1.3, across the Quinnipiac River, and the Chapel Street Bridge, mile 0.4, across the Mill River..., across the Quinnipiac River, and the Chapel Street Bridge, mile 0.4, across the Mill River, to reduce the...

  19. 76 FR 12 - Drawbridge Operation Regulations; New Haven Harbor, Quinnipiac and Mill Rivers, New Haven, CT

    Science.gov (United States)

    2011-01-03

    ... DEPARTMENT OF HOMELAND SECURITY Coast Guard 33 CFR Part 117 [Docket No. USCG-2010-1096] Drawbridge Operation Regulations; New Haven Harbor, Quinnipiac and Mill Rivers, New Haven, CT AGENCY: Coast Guard, DHS. ACTION: Notice of temporary deviation from regulations. SUMMARY: The Commander, First Coast Guard...

  20. Space Threat Warning: Foundation for Space Superiority, Avoiding a Space Pearl Harbor

    National Research Council Canada - National Science Library

    Burke, Alan W

    2006-01-01

    ... have stated the US must avoid a space Pearl Harbor. This concern is due to the idea that a decrease in the perceived threat to space assets after the demise of the Soviet Union coupled with a competition for space resources has resulted...

  1. Remembering Pearl Harbor: The USS Arizona Memorial. Teaching with Historic Places.

    Science.gov (United States)

    Vierra, John, Jr.

    This lesson describes and discusses the submerged remains of the battleship USS Arizona which rests on the silt of Pearl Harbor (Hawaii), just as it had settled on December 7, 1941, the day Japan attacked the U.S. fleet and began the Pacific battles of World War II. The lesson is based on the National Register of Historic Places registration file,…

  2. One surgeon's Army experience with "wound shock" from Pearl Harbor to the present.

    Science.gov (United States)

    Hardaway, Robert M

    2009-09-01

    The Army has had extensive experience in the study and treatment of shock, beginning with the American Civil War and continuing to the present. This is the story of one Army surgeon's experience, both in research and treatment of shock, from Pearl Harbor to the present.

  3. Japan's Monroe Doctrine?: Re-Framing the Story of Pearl Harbor

    Science.gov (United States)

    Murnane, John R.

    2007-01-01

    Ever since War Department propaganda films set the mold, historians have pretty much followed: the road to Pearl Harbor began in 1931 with Japanese aggression in Manchuria. Any "revision" regarding the War in the Pacific has its own set of problems--engendering controversy, often running up against patriotic sentiments and the tendency…

  4. Revisiting "Pearl Harbor": Resistance to Reel and Real Events in an English Language Classroom

    Science.gov (United States)

    Mackie, Ardiss; Norton, Bonny

    2006-01-01

    In this article, we draw on disruptive scenes in a postsecondary classroom to examine a critical incident concerning conflicting readings of the film "Pearl Harbor" (2001). We raise crucial questions for pedagogical work with popular film: Who speaks for whom about the meaning of a given film? Under what conditions do students resist…

  5. 78 FR 46855 - Safety Zone; East End Maritime Foundation Fireworks Display, Greenport Harbor, Greenport, NY

    Science.gov (United States)

    2013-08-02

    ... 1625-AA00 Safety Zone; East End Maritime Foundation Fireworks Display, Greenport Harbor, Greenport, NY... temporary rule proposes to establish a safety zone for the East End Maritime Foundation fireworks display....1. 0 2. Add Sec. 165.T01-0600 to read as follows: Sec. 165.T01-0600 Safety Zone; East End Maritime...

  6. 75 FR 18056 - Safety Zone; Fireworks Display, Patuxent River, Solomons Island Harbor, MD

    Science.gov (United States)

    2010-04-09

    ... the event, and enhancing public and maritime safety. Basis and Purpose Fireworks displays are... promote public and maritime safety during a fireworks display, and to protect mariners transiting the area...-AA00 Safety Zone; Fireworks Display, Patuxent River, Solomons Island Harbor, MD AGENCY: Coast Guard...

  7. 76 FR 57087 - Stone Harbor Investment Partners LP, et al.; Notice of Application

    Science.gov (United States)

    2011-09-15

    ... SECURITIES AND EXCHANGE COMMISSION [Investment Company Act Release No. 29784; File No. 812-13931] Stone Harbor Investment Partners LP, et al.; Notice of Application September 7, 2011. AGENCY: Securities..., or cause more than 10% of the acquired company's voting stock to be owned by investment companies and...

  8. 33 CFR 165.116 - Safety and Security Zones; Salem and Boston Harbors, Massachusetts.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Safety and Security Zones; Salem..., DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PORTS AND WATERWAYS SAFETY REGULATED NAVIGATION AREAS AND LIMITED... § 165.116 Safety and Security Zones; Salem and Boston Harbors, Massachusetts. (a) Location. The...

  9. 33 CFR 165.114 - Safety and Security Zones: Escorted Vessels-Boston Harbor, Massachusetts.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Safety and Security Zones... COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PORTS AND WATERWAYS SAFETY REGULATED NAVIGATION... Guard District § 165.114 Safety and Security Zones: Escorted Vessels—Boston Harbor, Massachusetts. (a...

  10. 78 FR 19158 - Safety Zone; USA Triathlon, Milwaukee Harbor, Milwaukee, WI

    Science.gov (United States)

    2013-03-29

    ... have minimal impact on the economy, will not interfere with other agencies, will not adversely alter... Milwaukee Harbor due to the 2013 and 2014 Olympic and Sprint Distance National Championships. This proposed... the 2013 and 2014 Olympic and Sprint Distance National Championships. DATES: Comments and related...

  11. Beyond Parasitism: Hepatic Lesions in Stranded Harbor Porpoises (Phocoena phocoena) Without Trematode (Campula oblonga) Infections

    NARCIS (Netherlands)

    S. Hiemstra; L. Harkema (Liesbeth); L.C.M. Wiersma (Lidewij); R.I. Keesler

    2015-01-01

    textabstractThe liver can be an indicator of the health of an individual or of a group, which can be especially important to identify agents that can cause disease in multiple species. To better characterize hepatic lesions in stranded harbor porpoises (Phocoena phocoena), we analyzed the livers

  12. Synergistic growth inhibition of cancer cells harboring the RET/PTC1 ...

    Indian Academy of Sciences (India)

    Synergistic growth inhibition of cancer cells harboring the RET/PTC1 oncogene by staurosporine and rotenone involves enhanced cell death. ANTÓNIO PEDRO GONÇALVES, ARNALDO VIDEIRA, VALDEMAR MÁXIMO and PAULA SOARES. J. Biosci. 36(4), September 2011, 639-648, © Indian Academy of Sciences.

  13. 33 CFR 117.669 - St. Louis River (Duluth Superior Harbor).

    Science.gov (United States)

    2010-07-01

    ... SECURITY BRIDGES DRAWBRIDGE OPERATION REGULATIONS Specific Requirements Minnesota § 117.669 St. Louis River (Duluth Superior Harbor). (a) The draws of the Burlington Northern railroad bridge, mile 5.7, shall open... Missabe and Iron Range Railway bridge, mile 16.3, need not be opened for the passage of vessels. The owner...

  14. 33 CFR 117.1083 - Duluth-Superior Harbor (St. Louis River).

    Science.gov (United States)

    2010-07-01

    ... SECURITY BRIDGES DRAWBRIDGE OPERATION REGULATIONS Specific Requirements Wisconsin § 117.1083 Duluth-Superior Harbor (St. Louis River). (a) The draws of the Burlington Northern railroad bridge, mile 5.7 at... blasts. (c) The draw of the Duluth, Missabe and Iron Range Railway bridge, mile 16.3 at Duluth, need not...

  15. 76 FR 33721 - Takes of Marine Mammals Incidental to Specified Activities; Harbor Activities Related to the...

    Science.gov (United States)

    2011-06-09

    ..., incidental to conducting Delta Mariner harbor operations for one year. NMFS reviewed the ULA application and... vitulina), and Northern elephant seals (Mirounga angustirostris) hauled out on Small Haul-out Site 1 to... lions; and 43 Northern elephant seals by Level B harassment only. Description of the Specified...

  16. 76 FR 21862 - Takes of Marine Mammals Incidental to Specified Activities; Harbor Activities Related to the...

    Science.gov (United States)

    2011-04-19

    ...; and 43 Northern elephant seals by Level B harassment only. To date, NMFS has issued eight, 1-year... elephant seals spend much of the year, generally about nine months, in the ocean. They are usually... Mariner harbor operations for one year. After addressing comments from NMFS, ULA modified its application...

  17. 77 FR 38587 - Takes of Marine Mammals Incidental to Specified Activities; Harbor Activities Related to the...

    Science.gov (United States)

    2012-06-28

    ... Northern elephant seals by Level B harassment only. To date, we have issued nine, one-year, Incidental... as far north as Alaska and as far south as Mexico. Northern elephant seals spend much of the year... numbers of marine mammals, incidental to conducting Delta Mariner harbor operations for one year. We...

  18. 78 FR 36662 - Safety Zone; Fairport Harbor Mardi Gras, Lake Erie, Fairport, OH

    Science.gov (United States)

    2013-06-19

    ... the Fairport Harbor Mardi Gras Fireworks display. This temporary safety zone is necessary to protect spectators and vessels from the hazards associated with a fireworks display. DATES: This rule is effective...: Temporary final rule. SUMMARY: The Coast Guard is establishing a temporary safety zone on Lake Erie...

  19. 33 CFR 334.470 - Cooper River and Charleston Harbor, S.C.; restricted areas.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Cooper River and Charleston....470 Cooper River and Charleston Harbor, S.C.; restricted areas. (a) The restricted areas. (1) Area No. 1 is that portion of the Cooper River beginning near the westerly shore north of Shipyard Creek at...

  20. 78 FR 18277 - Special Local Regulation; Low Country Splash, Wando River, Cooper River, and Charleston Harbor...

    Science.gov (United States)

    2013-03-26

    ...-AA08 Special Local Regulation; Low Country Splash, Wando River, Cooper River, and Charleston Harbor... proposes to issue a special local regulation on the waters of the Wando River, Cooper River, and Charleston... Country Splash is scheduled to take place on the waters of the Wando River, Cooper River, and Charleston...

  1. 33 CFR 165.709 - Security Zone; Charleston Harbor, Cooper River, South Carolina.

    Science.gov (United States)

    2010-07-01

    ..., Cooper River, South Carolina. 165.709 Section 165.709 Navigation and Navigable Waters COAST GUARD... § 165.709 Security Zone; Charleston Harbor, Cooper River, South Carolina. (a) Regulated area. The Coast Guard is establishing a fixed security zone on all waters of the Cooper River, bank-to-bank and surface...

  2. Novel B19-like parvovirus in the brain of a harbor seal

    NARCIS (Netherlands)

    R. Bodewes (Rogier); A.R. García (Ana Rubio); L.C.M. Wiersma (Lidewij); S. Getu (Sarah); M. Beukers (Martijn); C.M.E. Schapendonk (Claudia); P.R.W.A. van Run (Peter); M.W.G. van de Bildt (Marco); M.J. Poen (Marjolein); N. Osinga (Nynke); G.J. Sánchez Contreras (Guillermo); T. Kuiken (Thijs); S.L. Smits (Saskia); A.D.M.E. Osterhaus (Albert)

    2013-01-01

    textabstractUsing random PCR in combination with next-generation sequencing, a novel parvovirus was detected in the brain of a young harbor seal (Phoca vitulina) with chronic non-suppurative meningo-encephalitis that was rehabilitated at the Seal Rehabilitation and Research Centre (SRRC) in the

  3. 76 FR 82117 - Regulations Implementing the Longshore and Harbor Workers' Compensation Act: Recreational Vessels

    Science.gov (United States)

    2011-12-30

    ... activity, and whether it falls within the Coast Guard definition of a non- recreational vessel less than 20... recreational vessel work, the employee will be eligible for benefits based on the covered work. The last... Regulations Implementing the Longshore and Harbor Workers' Compensation Act: Recreational Vessels AGENCY...

  4. 75 FR 53574 - Safety Zone; Fireworks Displays, Potomac River, National Harbor, MD

    Science.gov (United States)

    2010-09-01

    ...-AA00 Safety Zone; Fireworks Displays, Potomac River, National Harbor, MD AGENCY: Coast Guard, DHS... safety of life on navigable waters during five fireworks displays launched from a discharge barge located... necessary to protect persons and vessels against the hazards associated with a fireworks display on...

  5. 78 FR 68002 - Safety Zone for Fireworks Display, Baltimore Harbor, Baltimore, MD

    Science.gov (United States)

    2013-11-13

    ... 1625-AA00 Safety Zone for Fireworks Display, Baltimore Harbor, Baltimore, MD AGENCY: Coast Guard, DHS... safety of life on navigable waters during a fireworks display launched from a barge located in Baltimore... rule involves a fireworks display associated with a New Year's Eve event that will take place in...

  6. 33 CFR 110.197 - Galveston Harbor, Bolivar Roads Channel, Texas.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Galveston Harbor, Bolivar Roads Channel, Texas. 110.197 Section 110.197 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF... Roads Channel, Texas. (a)(1) Anchorage area (A). The water bounded by a line connecting the following...

  7. 78 FR 57319 - Children's Online Privacy Protection Rule Safe Harbor Proposed Self-Regulatory Guidelines...

    Science.gov (United States)

    2013-09-18

    ...-AB20 Children's Online Privacy Protection Rule Safe Harbor Proposed Self-Regulatory Guidelines; kidSAFE... proposed self-regulatory guidelines submitted by the kidSAFE Seal Program (``kidSAFE''), owned and operated... enabling industry groups or others to submit to the Commission for approval self-regulatory guidelines that...

  8. 77 FR 11434 - Safety Zone; Patapsco River, Northwest and Inner Harbors, Baltimore, MD

    Science.gov (United States)

    2012-02-27

    ...-AA00 Safety Zone; Patapsco River, Northwest and Inner Harbors, Baltimore, MD AGENCY: Coast Guard, DHS... intended route immediately prior to, during, and following the scheduled towing evolution, vessel traffic... made to the maritime community via marine information broadcasts so mariners may adjust their plans...

  9. 75 FR 5907 - Safety Zone; Dive Platform, Pago Pago Harbor, American Samoa

    Science.gov (United States)

    2010-02-05

    ... platform vessel in Pago Pago Harbor, American Samoa, while diving operations are under way in and around the CHEHALIS wreck. The safety zone is necessary to protect other vessels and the general public from... Pago, American Samoa. Today, the CHEHALIS wreck remains a potential pollution threat to the environment...

  10. 33 CFR 334.1420 - Pacific Ocean off Orote Point, Apra Harbor, Island of Guam, Marianas Islands; small arms firing...

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Pacific Ocean off Orote Point, Apra Harbor, Island of Guam, Marianas Islands; small arms firing range. 334.1420 Section 334.1420... AND RESTRICTED AREA REGULATIONS § 334.1420 Pacific Ocean off Orote Point, Apra Harbor, Island of Guam...

  11. 33 CFR 334.85 - New York Harbor, adjacent to the Stapleton Naval Station, Staten Island, New York; restricted area.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false New York Harbor, adjacent to the Stapleton Naval Station, Staten Island, New York; restricted area. 334.85 Section 334.85 Navigation and... RESTRICTED AREA REGULATIONS § 334.85 New York Harbor, adjacent to the Stapleton Naval Station, Staten Island...

  12. 33 CFR 165.776 - Security Zone; Coast Guard Base San Juan, San Juan Harbor, Puerto Rico

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Security Zone; Coast Guard Base San Juan, San Juan Harbor, Puerto Rico 165.776 Section 165.776 Navigation and Navigable Waters COAST... Guard District § 165.776 Security Zone; Coast Guard Base San Juan, San Juan Harbor, Puerto Rico (a...

  13. 33 CFR 334.840 - Waters of Lake Michigan south of Northerly Island at entrance to Burnham Park Yacht Harbor...

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Waters of Lake Michigan south of Northerly Island at entrance to Burnham Park Yacht Harbor, Chicago, Ill.; danger zone adjacent to airport on... Michigan south of Northerly Island at entrance to Burnham Park Yacht Harbor, Chicago, Ill.; danger zone...

  14. 78 FR 16208 - Safety Zone; V. I. Carnival Finale; St. Thomas Harbor; St. Thomas, U.S. Virgin Islands

    Science.gov (United States)

    2013-03-14

    ... 1625-AA00 Safety Zone; V. I. Carnival Finale; St. Thomas Harbor; St. Thomas, U.S. Virgin Islands AGENCY... establish a safety zone on the waters of St. Thomas Harbor in St. Thomas, U. S. Virgin Islands during the V... between 9 a.m. and 5 p.m., Monday through Friday, except federal holidays. The telephone number is 202-366...

  15. 78 FR 16211 - Safety Zone, Corp. Event Finale UHC, St. Thomas Harbor; St. Thomas, U.S. Virgin Islands

    Science.gov (United States)

    2013-03-14

    ... 1625-AA00 Safety Zone, Corp. Event Finale UHC, St. Thomas Harbor; St. Thomas, U.S. Virgin Islands... establish a temporary safety zone on the waters of St. Thomas Harbor in St. Thomas, U.S. Virgin Islands... through Friday, except federal holidays. The telephone number is 202-366-9329. See the ``Public...

  16. 78 FR 23489 - Safety Zone; V.I. Carnival Finale, St. Thomas Harbor; St. Thomas, U.S.V.I.

    Science.gov (United States)

    2013-04-19

    ... waters of St. Thomas Harbor in St. Thomas, U.S. Virgin Islands during the V.I. Carnival Finale, a... through Friday, except Federal holidays. FOR FURTHER INFORMATION CONTACT: If you have questions on this... event. The event will be held on the waters of St. Thomas Harbor, St. Thomas, U. S. Virgin Islands...

  17. 78 FR 7663 - SLR; 2013 International Rolex Regatta; St. Thomas Harbor; St. Thomas, U.S. Virgin Islands

    Science.gov (United States)

    2013-02-04

    ... 1625-AA08 SLR; 2013 International Rolex Regatta; St. Thomas Harbor; St. Thomas, U.S. Virgin Islands... special local regulations on the waters of St. Thomas Harbor in St. Thomas, U. S. Virgin Islands during... 5 p.m., Monday through Friday, except federal holidays. The telephone number is 202-366-9329. See...

  18. 77 FR 60319 - Harbor Porpoise Take Reduction Plan; Coastal Gulf of Maine Closure Area Established With a...

    Science.gov (United States)

    2012-10-03

    ...-XC099 Harbor Porpoise Take Reduction Plan; Coastal Gulf of Maine Closure Area Established With a... Atmospheric Administration (NOAA), Commerce. ACTION: Establishment of the Coastal Gulf of Maine Closure Area... Service (NMFS) announces the establishment of the Coastal Gulf of Maine Closure Area under the Harbor...

  19. 77 FR 47331 - Regulated Navigation Area-New Haven Harbor, Quinnipiac River, Mill River, New Haven, CT; Pearl...

    Science.gov (United States)

    2012-08-08

    ...-AA11 Regulated Navigation Area--New Haven Harbor, Quinnipiac River, Mill River, New Haven, CT; Pearl... navigable waters of New Haven Harbor, Quinnipiac River and Mill River. The current RNA pertains only to the..., Quinnipiac River, and Mill River RNA. The proposed amendment would give the Captain of the Port Sector Long...

  20. 33 CFR 162.260 - Channel leading to San Juan Harbor, P.R.; use, administration, and navigation.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Channel leading to San Juan Harbor, P.R.; use, administration, and navigation. 162.260 Section 162.260 Navigation and Navigable... WATERWAYS NAVIGATION REGULATIONS § 162.260 Channel leading to San Juan Harbor, P.R.; use, administration...

  1. A Comparison of Microbial Water Quality and Diversity for Ballast and Tropical Harbor Waters.

    Science.gov (United States)

    Ng, Charmaine; Le, Thai-Hoang; Goh, Shin Giek; Liang, Liang; Kim, Yiseul; Rose, Joan B; Yew-Hoong, Karina Gin

    2015-01-01

    Indicator organisms and antibiotic resistance were used as a proxy to measure microbial water quality of ballast tanks of ships, and surface waters in a tropical harbor. The survival of marine bacteria in ballast tanks appeared to diminish over longer water retention time, with a reduction of cell viability observed after a week based on heterotrophic plate counts. Pyrosequencing of 16S rRNA genes showed distinct differences in microbial composition of ballast and harbor waters. The harbor waters had a higher abundance of operational taxonomic units (OTUs) assigned to Cyanobacteria (Synechococcus spp.) and α-proteobacteria (SAR11 members), while marine hydrocarbon degraders such as γ-proteobacteria (Ocenspirillaes spp., Thiotrchales spp.) and Bacteroidetes (Flavobacteriales spp.) dominated the ballast water samples. Screening of indicator organisms found Escherichia coli (E. coli), Enterococcus and Pseudomonas aeruginosa (P. aeruginosa) in two or more of the ballast and harbor water samples tested. Vibrio spp. and Salmonella spp. were detected exclusively in harbor water samples. Using quantitative PCR (qPCR), we screened for 13 antibiotic resistant gene (ARG) targets and found higher abundances of sul1 (4.13-3.44 x 102 copies/mL), dfrA (0.77-1.80 x10 copies/mL) and cfr (2.00-5.21 copies/mL) genes compared to the other ARG targets selected for this survey. These genes encode for resistance to sulfonamides, trimethoprim and chloramphenicol-florfenicol antibiotics, which are also known to persist in sediments of aquaculture farms and coastal environments. Among the ARGs screened, we found significant correlations (Pwater quality survey, quantitatively assessing indicators of antibiotic resistance, potentially pathogenic organisms and a broad-brush description of difference in microbial composition and diversity between open oceans and tropical coastal environments through the use of next generation sequencing technology.

  2. Distribution and contamination status of chromium in surface sediments of northern Kaohsiung Harbor, Taiwan.

    Science.gov (United States)

    Dong, Cheng-Di; Chen, Chiu-Wen; Chen, Chih-Feng

    2013-07-01

    The distribution, enrichment, accumulation, and potential ecological risk of chromium (Cr) in the surface sediments of northern Kaohsiung Harbor, Taiwan, China were investigated. Sediment samples from ten locations located between the river mouths and harbor entrance of northern Kaohsiung Harbor were collected quarterly in 2011 and characterized for Cr, aluminum, water content, organic matter, total nitrogen, total phosphorous, total grease, and grain size. Results showed that the Cr concentrations varied from 27.0 to 361.9 mg/kg with an average of (113.5 +/- 87.0) mg/kg. High Cr concentration was observed near the Jen-Gen River mouth. The mean Cr concentration was high at 255.5 mg/kg, which was at least 2 to 7 times than that of other sites. This might imply significant Cr contribution from upstream receiving tanneries wastewater into the Jen-Gen River. The spatial distribution of Cr reveals relatively high in the river mouth region, especially in Jen-Gen River, and gradually diminishes toward the harbor entrance region. This indicates that the major sources of Cr pollution from upstream industrial and municipal wastewaters discharged along the river bank; and Cr may drift with sea current and be dispersed into open sea. Moreover, Cr concentrations correlated closely to the physical-chemical properties of the sediments, which suggested the influence of industrial and municipal wastewaters discharged from the neighboring industrial parks and river basins. Results from the enrichment factor and geo-accumulation index analyses imply that the Jen-Gen River sediments can be characterized as moderate enrichment and none to medium accumulation of Cr, respectively. However, results of potential ecological risk index indicate that the sediment has low ecological potential risk. The results can provide valuable information to developing future strategies for the management of river mouth and harbor.

  3. Current and historic mercury deposition to New Haven Harbor (CT, USA): Implications for industrial coastal environments

    International Nuclear Information System (INIS)

    Clark, Heather F.; Benoit, Gaboury

    2009-01-01

    This study quantifies historic and current mercury contamination in New Haven Harbor (New Haven, Connecticut, USA) through the analysis of sediment cores. The mercury concentration measured in surface sediment ranged from 320 to 1640 μg kg -1 with an average of 530 μg kg -1 . The harbor is relatively small in area (6.6 km 2 ) but displays a large range in concentrations, illustrating the important methodological issue that a large number of samples may be necessary to capture the variability in even a small area. Depth profiles of mercury reflect sedimentation over a range of 20 to 200 years and indicate a complex history of contamination. Mercury depth profiles were compared with lead, copper, cadmium, and silver concentrations and the metals generally covary. This trend indicates that the sources of mercury and heavy metals are linked and that regionally specific sources dominate the historic input of metals rather than large-scale atmospheric deposition patterns. Results also show there are large differences in absolute concentrations of metals among sites in the harbor. Differences in the abundance of Fe-rich, fine-grained sediment likely control the level of metals in various parts of the harbor. Proximity to current sources and the long, diverse industrial history of the harbor also influence the distribution pattern. All of the cores can be modeled as mixing between pre-industrial sediments and either one or two pollution endmembers. This study demonstrates the importance of riverine sources in the mass balance of mercury delivered to coastal areas and of watershed management to preserve coastal ecosystems.

  4. APRILab: Regulation Dilemma in South Harbor & Aalborg East, Denmark

    DEFF Research Database (Denmark)

    Hansen, Jesper Rohr

    Copenhagen [in English, ‘South Harbour’] demonstrates an urban periphery development that in terms of strategy formation, project development and implementation has been running since the late 1990s. Accordingly, most of the city district of Sydhavn Copenhagen has been planned for. However, despite...... is the financial crisis that paralyzed the development in the area in the years 2007-2011; however, as this report shows, the deployment of new regulative tools as well as an expansionist municipal growth strategy have also contributed to this slow progress, generating problems of cooperation between landowners...... and diverting the attention and resources of the municipal system (administratively, politically). The suburban case, Aalborg Øst [in English, ‘Aalborg East’], in the City of Aalborg, is another type of case completely. This case is immature in the sense that only the first building blocks for a strategy...

  5. Nonindigenous Marine Species Introductions in the harbors of the South and West Shores of Oahu, Hawaii 1997-1998 (NODC Accession 0000324)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Despite the potential importance of Honolulu Harbor or other commercial harbors on Oahu as potential gateways for nonindigenous marine species to enter the Hawaiian...

  6. Effects of caffeine on circadian phase, amplitude and period evaluated in cells in vitro and peripheral organs in vivo in PER2::LUCIFERASE mice

    Science.gov (United States)

    Narishige, Seira; Kuwahara, Mari; Shinozaki, Ayako; Okada, Satoshi; Ikeda, Yuko; Kamagata, Mayo; Tahara, Yu; Shibata, Shigenobu

    2014-01-01

    Background and Purpose Caffeine is one of the most commonly used psychoactive substances. Circadian rhythms consist of the main suprachiasmatic nucleus (SCN) clocks and peripheral clocks. Although caffeine lengthens circadian rhythms and modifies phase changes in SCN-operated rhythms, the effects on caffeine on the phase, period and amplitude of peripheral organ clocks are not known. In addition, the role of cAMP/Ca2+ signalling in effects of caffeine on rhythm has not been fully elucidated. Experimental Approach We examined whether chronic or transient application of caffeine affects circadian period/amplitude and phase by evaluating bioluminescence rhythm in PER2::LUCIFERASE knock-in mice. Circadian rhythms were monitored in vitro using fibroblasts and ex vivo and in vivo for monitoring of peripheral clocks. Key Results Chronic application of caffeine (0.1–10 mM) increased period and amplitude in vitro. Transient application of caffeine (10 mM) near the bottom of the decreasing phase of bioluminescence rhythm caused phase advance in vitro. Caffeine (0.1%) intake caused a phase delay under light–dark or constant dark conditions, suggesting a period-lengthening effect in vivo. Caffeine (20 mg·kg−1) at daytime or at late night-time caused phase advance or delay in bioluminescence rhythm in the liver and kidney respectively. The complicated roles of cAMP/Ca2+ signalling may be involved in the caffeine-induced increase of period and amplitude in vitro. Conclusions and Implications Caffeine affects circadian rhythm in mice by lengthening the period and causing a phase shift of peripheral clocks. These results suggest that caffeine intake with food/drink may help with food-induced resetting of peripheral circadian clocks. PMID:25160990

  7. Experimental Evolution of Escherichia coli Harboring an Ancient Translation Protein.

    Science.gov (United States)

    Kacar, Betül; Ge, Xueliang; Sanyal, Suparna; Gaucher, Eric A

    2017-03-01

    The ability to design synthetic genes and engineer biological systems at the genome scale opens new means by which to characterize phenotypic states and the responses of biological systems to perturbations. One emerging method involves inserting artificial genes into bacterial genomes and examining how the genome and its new genes adapt to each other. Here we report the development and implementation of a modified approach to this method, in which phylogenetically inferred genes are inserted into a microbial genome, and laboratory evolution is then used to examine the adaptive potential of the resulting hybrid genome. Specifically, we engineered an approximately 700-million-year-old inferred ancestral variant of tufB, an essential gene encoding elongation factor Tu, and inserted it in a modern Escherichia coli genome in place of the native tufB gene. While the ancient homolog was not lethal to the cell, it did cause a twofold decrease in organismal fitness, mainly due to reduced protein dosage. We subsequently evolved replicate hybrid bacterial populations for 2000 generations in the laboratory and examined the adaptive response via fitness assays, whole genome sequencing, proteomics, and biochemical assays. Hybrid lineages exhibit a general adaptive strategy in which the fitness cost of the ancient gene was ameliorated in part by upregulation of protein production. Our results suggest that an ancient-modern recombinant method may pave the way for the synthesis of organisms that exhibit ancient phenotypes, and that laboratory evolution of these organisms may prove useful in elucidating insights into historical adaptive processes.

  8. Rain-shadow: An area harboring "Gray Ocean" clouds

    Science.gov (United States)

    Padmakumari, B.; Maheskumar, R. S.; Harikishan, G.; Morwal, S. B.; Kulkarni, J. R.

    2018-06-01

    The characteristics of monsoon convective clouds over the rain-shadow region of north peninsular India have been investigated using in situ aircraft cloud microphysical observations collected during Cloud Aerosol Interaction and Precipitation Enhancement EXperiment (CAIPEEX). The parameters considered for characterization are: liquid water content (LWC), cloud vertical motion (updraft, downdraft: w), cloud droplet number concentration (CDNC) and effective radius (Re). The results are based on 15 research flights which were conducted from the base station Hyderabad during summer monsoon season. The clouds studied were developing congestus. The clouds have low CDNC and low updraft values resembling the oceanic convective clouds. The super-saturation in clouds is found to be low (≤0.2%) due to low updrafts. The land surface behaves like ocean surface during monsoon as deduced from Bowen ratio. Microphysically the clouds showed oceanic characteristics. However, these clouds yield low rainfall due to their low efficiency (mean 14%). The cloud parameters showed a large variability; hence their characteristic values are reported in terms of median values. These values will serve the numerical models for rainfall simulations over the region and also will be useful as a scientific basis for cloud seeding operations to increase the rainfall efficiency. The study revealed that monsoon convective clouds over the rain-shadow region are of oceanic type over the gray land, and therefore we christen them as "Gray Ocean" clouds.

  9. A Probabilistic and Observation Based Methodology to Estimate Small Craft Harbor Vulnerability to Tsunami Events

    Science.gov (United States)

    Keen, A. S.; Lynett, P. J.; Ayca, A.

    2016-12-01

    Because of the damage resulting from the 2010 Chile and 2011 Japanese tele-tsunamis, the tsunami risk to the small craft marinas in California has become an important concern. The talk will outline an assessment tool which can be used to assess the tsunami hazard to small craft harbors. The methodology is based on the demand and structural capacity of the floating dock system, composed of floating docks/fingers and moored vessels. The structural demand is determined using a Monte Carlo methodology. Monte Carlo methodology is a probabilistic computational tool where the governing might be well known, but the independent variables of the input (demand) as well as the resisting structural components (capacity) may not be completely known. The Monte Carlo approach uses a distribution of each variable, and then uses that random variable within the described parameters, to generate a single computation. The process then repeats hundreds or thousands of times. The numerical model "Method of Splitting Tsunamis" (MOST) has been used to determine the inputs for the small craft harbors within California. Hydrodynamic model results of current speed, direction and surface elevation were incorporated via the drag equations to provide the bases of the demand term. To determine the capacities, an inspection program was developed to identify common features of structural components. A total of six harbors have been inspected ranging from Crescent City in Northern California to Oceanside Harbor in Southern California. Results from the inspection program were used to develop component capacity tables which incorporated the basic specifications of each component (e.g. bolt size and configuration) and a reduction factor (which accounts for the component reduction in capacity with age) to estimate in situ capacities. Like the demand term, these capacities are added probabilistically into the model. To date the model has been applied to Santa Cruz Harbor as well as Noyo River. Once

  10. Colistin- and Carbapenem-Resistant Escherichia coli Harboring mcr-1 and blaNDM-5, Causing a Complicated Urinary Tract Infection in a Patient from the United States.

    Science.gov (United States)

    Mediavilla, José R; Patrawalla, Amee; Chen, Liang; Chavda, Kalyan D; Mathema, Barun; Vinnard, Christopher; Dever, Lisa L; Kreiswirth, Barry N

    2016-08-30

    Colistin is increasingly used as an antibiotic of last resort for the treatment of carbapenem-resistant Gram-negative infections. The plasmid-borne colistin resistance gene mcr-1 was initially identified in animal and clinical samples from China and subsequently reported worldwide, including in the United States. Of particular concern is the spread of mcr-1 into carbapenem-resistant bacteria, thereby creating strains that approach pan-resistance. While several reports of mcr-1 have involved carbapenem-resistant strains, no such isolates have been described in the United States. Here, we report the isolation and identification of an Escherichia coli strain harboring both mcr-1 and carbapenemase gene blaNDM-5 from a urine sample in a patient without recent travel outside the United States. The isolate exhibited resistance to both colistin and carbapenems, but was susceptible to amikacin, aztreonam, gentamicin, nitrofurantoin, tigecycline, and trimethoprim-sulfamethoxazole. The mcr-1- and blaNDM-5-harboring plasmids were completely sequenced and shown to be highly similar to plasmids previously reported from China. The strain in this report was first isolated in August 2014, highlighting an earlier presence of mcr-1 within the United States than previously recognized. Colistin has become the last line of defense for the treatment of infections caused by Gram-negative bacteria resistant to multiple classes of antibiotics, in particular carbapenem-resistant Enterobacteriaceae (CRE). Resistance to colistin, encoded by the plasmid-borne gene mcr-1, was first identified in animal and clinical samples from China in November 2015 and has subsequently been reported from numerous other countries. In April 2016, mcr-1 was identified in a carbapenem-susceptible Escherichia coli strain from a clinical sample in the United States, followed by a second report from a carbapenem-susceptible E. coli strain originally isolated in May 2015. We report the isolation and identification of

  11. Pediatric fecal microbiota harbor diverse and novel antibiotic resistance genes.

    Directory of Open Access Journals (Sweden)

    Aimée M Moore

    Full Text Available Emerging antibiotic resistance threatens human health. Gut microbes are an epidemiologically important reservoir of resistance genes (resistome, yet prior studies indicate that the true diversity of gut-associated resistomes has been underestimated. To deeply characterize the pediatric gut-associated resistome, we created metagenomic recombinant libraries in an Escherichia coli host using fecal DNA from 22 healthy infants and children (most without recent antibiotic exposure, and performed functional selections for resistance to 18 antibiotics from eight drug classes. Resistance-conferring DNA fragments were sequenced (Illumina HiSeq 2000, and reads assembled and annotated with the PARFuMS computational pipeline. Resistance to 14 of the 18 antibiotics was found in stools of infants and children. Recovered genes included chloramphenicol acetyltransferases, drug-resistant dihydrofolate reductases, rRNA methyltransferases, transcriptional regulators, multidrug efflux pumps, and every major class of beta-lactamase, aminoglycoside-modifying enzyme, and tetracycline resistance protein. Many resistance-conferring sequences were mobilizable; some had low identity to any known organism, emphasizing cryptic organisms as potentially important resistance reservoirs. We functionally confirmed three novel resistance genes, including a 16S rRNA methylase conferring aminoglycoside resistance, and two tetracycline-resistance proteins nearly identical to a bifidobacterial MFS transporter (B. longum s. longum JDM301. We provide the first report to our knowledge of resistance to folate-synthesis inhibitors conferred by a predicted Nudix hydrolase (part of the folate synthesis pathway. This functional metagenomic survey of gut-associated resistomes, the largest of its kind to date, demonstrates that fecal resistomes of healthy children are far more diverse than previously suspected, that clinically relevant resistance genes are present even without recent selective

  12. The Arabidopsis thaliana proteome harbors undiscovered multi-domain molecules with functional guanylyl cyclase catalytic centers

    KAUST Repository

    Wong, Aloysius Tze

    2013-07-08

    Background: Second messengers link external cues to complex physiological responses. One such messenger, 3\\',5\\'-cyclic guanosine monophosphate (cGMP), has been shown to play a key role in many physiological responses in plants. However, in higher plants, guanylyl cyclases (GCs), enzymes that generate cGMP from guanosine-5\\'-triphosphate (GTP) have remained elusive until recently. GC search motifs constructed from the alignment of known GCs catalytic centers form vertebrates and lower eukaryotes have led to the identification of a number of plant GCs that have been characterized in vitro and in vivo.Presentation of the hypothesis.Recently characterized GCs in Arabidopsis thaliana contributed to the development of search parameters that can identify novel candidate GCs in plants. We hypothesize that there are still a substantial number (> 40) of multi-domain molecules with potentially functional GC catalytic centers in plants that remain to be discovered and characterized. Testing the hypothesis. The hypothesis can be tested, firstly, by computational methods constructing 3D models of selected GC candidates using available crystal structures as templates. Homology modeling must include substrate docking that can provide support for the structural feasibility of the GC catalytic centers in those candidates. Secondly, recombinant peptides containing the GC domain need to be tested in in vitro GC assays such as the enzyme-linked immune-sorbent assay (ELISA) and/or in mass spectrometry based cGMP assays. In addition, quantification of in vivo cGMP transients with fluorescent cGMP-reporter assays in wild-type or selected mutants will help to elucidate the biological role of novel GCs.Implications of the hypothesis.If it turns out that plants do harbor a large number of functional GC domains as part of multi-domain enzymes, then major new insights will be gained into the complex signal transduction pathways that link cGMP to fundamental processes such as ion transport

  13. Detection of estrogenic activity in sediment-associated compounds using in vitro reporter gene assays

    NARCIS (Netherlands)

    Legler, J.; Dennekamp, M.; Vethaak, A.D.; Brouwer, A.; Koeman, J.H.; Burg, van der B.; Murk, A.J.

    2002-01-01

    Sediments may be the ultimate sink for persistent (xeno-) estrogenic compounds released into the aquatic environment. Sediment-associated estrogenic potency was measured with an estrogen receptor-mediated luciferase reporter gene (ER-CALUX) assay and compared with a recombinant yeast screen. The

  14. A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes

    DEFF Research Database (Denmark)

    Seefried, Lothar; Mueller-Deubert, Sigrid; Wentzer, Thomas Schwarz

    2010-01-01

    of mechanotransduction and its crosstalk with biochemically induced signal transduction, AP1 and SP1 luciferase reporter gene constructs were cloned and transfected into various cell lines and primary cells. A newly developed bioreactor and small-scale 24-well polyurethane dishes were used to apply cyclic stretching...

  15. Effects of Epigenetic Modulation on Reporter Gene Expression: Implications for Stem Cell Imaging

    Science.gov (United States)

    Krishnan, Manickam; Park, Jinha M.; Cao, Feng; Wang, Dongxu; Paulmurugan, Ramasay; Tseng, Jeffrey R.; Gonzalgo, Mark L.; Gambhir, Sanjiv S.; Wu, Joseph C.

    2013-01-01

    Tracking stem cell localization, survival, differentiation, and proliferation following transplantation in living subjects is essential for understanding stem cell biology and physiology. In this study, we investigated the long-term stability of reporter gene expression in an embryonic rat cardiomyoblast cell line and the role of epigenetic modulation on reversing reporter gene silencing. Cells were stably transfected with plasmids carrying cytomegalovirus promoter driving firefly luciferase reporter gene (CMV-Fluc) and passaged repeatedly for 3–8 months. Within the highest expressor clone, the firefly luciferase activity decreased progressively from passage-1 (843±28) to passage-20 (250±10) to passage-40 (44±3) to passage-60 (3±1 RLU/µg) (P<0.05 vs. passage-1). Firefly luciferase activity was maximally rescued by treatment with 5-azacytidine (DNA methyltransferase inhibitor) compared to trichostatin A (histone deacetylase inhibitor) and retinoic acid (transcriptional activator) (P<0.05). Increasing dosages of 5-azacytidine treatment led to higher levels of firefly luciferase mRNA (RT-PCR) and protein (Western blots) and inversely lower levels of methylation in the CMV promoter (DNA nucleotide sequence). These in vitro results were extended to in vivo bioluminescence imaging (BLI) of cell transplant in living animals. Cells treated with 5-azacytidine were monitored for 2 weeks compared to 1 week for untreated cells (P<0.05). These findings should have important implications for reporter gene-based imaging of stem cell transplantation. PMID:16246867

  16. pLIVE-EGFP: A liver specific vector carrying the EGFP reporter for ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-19

    Oct 19, 2009 ... system for monitoring gene expression, intracellular protein trafficking or localization. Several types of reporter systems, such as secretory alkaline phosphatase (SEAP), luciferase, β-galactosidase, or fluorescent proteins (FPs), have been widely applied in these researches (Naylor,. 1999; Branchini, et al., ...

  17. Hydraulic experiment on flow and topography change in harbor due to tsunami and its numerical simulation

    International Nuclear Information System (INIS)

    Fujii, Naoki; Ikeno, Masaaki; Sakakiyama, Tsutomu; Matsuyama, Masafumi; Takao, Makoto; Mukohara, Takeshi

    2009-01-01

    Numerical model of topography change is important to examine collapse of the harbor facilities by sand transport due to tsunami. Problems for evaluation of sand transport due to tsunami with topography change model are in precision of the numerical model and topography change data. Therefore, we installed the harbor in large-scaled wave tank and carried out experiment about tsunami flow and topography change to get those detailed data. For results provided by experimental test, we applied the topography change model of Ikeno et al. (2009a) and evaluated it about the reproduction characteristics. As a result, it was confirmed that reproduction of an experiment improved by using new pickup rate formula proposed by Ikeno et al. (2009a). (author)

  18. Bacterial Communities in Polluted Seabed Sediments: A Molecular Biology Assay in Leghorn Harbor

    Directory of Open Access Journals (Sweden)

    Carolina Chiellini

    2013-01-01

    Full Text Available Seabed sediments of commercial ports are often characterized by high pollution levels. Differences in number and distribution of bacteria in such areas can be related to distribution of pollutants in the port and to sediment conditions. In this study, the bacterial communities of five sites from Leghorn Harbor seabed were characterized, and the main bacterial groups were identified. T-RFLP was used for all samples; two 16S rRNA libraries and in silico digestion of clones were used to identify fingerprint profiles. Library data, phylogenetic analysis, and T-RFLP coupled with in silico digestion of the obtained sequences evidenced the dominance of Proteobacteria and the high percentage of Bacteroidetes in all sites. The approach highlighted similar bacterial communities between samples coming from the five sites, suggesting a modest differentiation among bacterial communities of different harbor seabed sediments and hence the capacity of bacterial communities to adapt to different levels and types of pollution.

  19. Assay dependence of Brucella antibody prevalence in a declining Alaskan harbor seal (Phoca vitulina population

    Directory of Open Access Journals (Sweden)

    Hueffer Karsten

    2013-01-01

    Full Text Available Abstract Background Brucella is a group of bacteria that causes brucellosis, which can affect population health and reproductive success in many marine mammals. We investigated the serological prevalence of antibodies against Brucella bacteria in a declining harbor seal population in Glacier Bay National Park, Alaska. Results Prevalence ranged from 16 to 74 percent for those tests detecting antibodies, indicating that harbor seals in Glacier Bay have been exposed to Brucella bacteria. However, the actual level of serological prevalence could not be determined because results were strongly assay-dependent. Conclusions This study reinforces the need to carefully consider assay choice when comparing different studies on the prevalence of anti–Brucella antibodies in pinnipeds and further highlights the need for species- or taxon-specific assay validation for both pathogen and host species.

  20. Multimodality imaging of reporter gene expression using a novel fusion vector in living cells and animals

    Science.gov (United States)

    Gambhir, Sanjiv [Portola Valley, CA; Pritha, Ray [Mountain View, CA

    2011-06-07

    Novel double and triple fusion reporter gene constructs harboring distinct imagable reporter genes are provided, as well as applications for the use of such double and triple fusion constructs in living cells and in living animals using distinct imaging technologies.

  1. Re-alimentation in harbor seal pups: effects on the somatotropic axis and growth rate.

    Science.gov (United States)

    Richmond, Julie P; Norris, Tenaya; Zinn, Steven A

    2010-01-15

    The metabolic hormones, growth hormone (GH) and insulin-like growth factor (IGF)-I, together with IGF binding proteins (IGFBP), have been well studied in domestic species and are the primary components of the somatotropic axis. This hormone axis is responsive to nutrient intake, associated with growth rate, and accretion of protein and adipose. However, this relationship has not been evaluated in species that rely heavily on adipose stores for survival, such as pinnipeds. The primary objectives of this research were to investigate the response of the somatotropic axis to reduced nutrient intake and re-alimentation in rehabilitated harbor seal pups, and to assess if these hormones are related to nutritional status and growth rate in harbor seals. Stranded harbor seal pups (n=24) arrived at the rehabilitation facility very thin after fasting for several days (nutritional nadir). Throughout rehabilitation nutrient intake increased and pups gained mass and body condition. Concentrations of GH and IGFBP-2 decreased with re-alimentation, while IGF-I and IGFBP-3 concentrations increased. Overall, GH and IGFBP-2 were negatively associated and IGF-I and IGFBP-3 were positively associated with growth rate and increased body condition of harbor sea pups. Further, the magnitude of the growth response was related to the magnitude in response of the somatotropic axis to varied levels of intake. These data suggest that multiple components of the somatotropic axis may be used to assess the energy status of individuals and may also provide information on the level of feed intake that is predictive of growth rate.

  2. Female harbor seal (Phoca vitulina) behavioral response to playbacks of underwater male acoustic advertisement displays.

    Science.gov (United States)

    Matthews, Leanna P; Blades, Brittany; Parks, Susan E

    2018-01-01

    During the breeding season, male harbor seals ( Phoca vitulina ) make underwater acoustic displays using vocalizations known as roars. These roars have been shown to function in territory establishment in some breeding areas and have been hypothesized to be important for female choice, but the function of these sounds remains unresolved. This study consisted of a series of playback experiments in which captive female harbor seals were exposed to recordings of male roars to determine if females respond to recordings of male vocalizations and whether or not they respond differently to roars from categories with different acoustic characteristics. The categories included roars with characteristics of dominant males (longest duration, lowest frequency), subordinate males (shortest duration, highest frequency), combinations of call parameters from dominant and subordinate males (long duration, high frequency and short duration, low frequency), and control playbacks of water noise and water noise with tonal signals in the same frequency range as male signals. Results indicate that overall females have a significantly higher level of response to playbacks that imitate male vocalizations when compared to control playbacks of water noise. Specifically, there was a higher level of response to playbacks representing dominant male vocalization when compared to the control playbacks. For most individuals, there was a greater response to playbacks representing dominant male vocalizations compared to playbacks representing subordinate male vocalizations; however, there was no statistical difference between those two playback types. Additionally, there was no difference between the playbacks of call parameter combinations and the controls. Investigating female preference for male harbor seal vocalizations is a critical step in understanding the harbor seal mating system and further studies expanding on this captive study will help shed light on this important issue.

  3. Calculating background levels for ecological risk parameters in toxic harbor sediment

    Science.gov (United States)

    Leadon, C.J.; McDonnell, T.R.; Lear, J.; Barclift, D.

    2007-01-01

    Establishing background levels for biological parameters is necessary in assessing the ecological risks from harbor sediment contaminated with toxic chemicals. For chemicals in sediment, the term contaminated is defined as having concentrations above background and significant human health or ecological risk levels. For biological parameters, a site could be considered contaminated if levels of the parameter are either more or less than the background level, depending on the specific parameter. Biological parameters can include tissue chemical concentrations in ecological receptors, bioassay responses, bioaccumulation levels, and benthic community metrics. Chemical parameters can include sediment concentrations of a variety of potentially toxic chemicals. Indirectly, contaminated harbor sediment can impact shellfish, fish, birds, and marine mammals, and human populations. This paper summarizes the methods used to define background levels for chemical and biological parameters from a survey of ecological risk investigations of marine harbor sediment at California Navy bases. Background levels for regional biological indices used to quantify ecological risks for benthic communities are also described. Generally, background stations are positioned in relatively clean areas exhibiting the same physical and general chemical characteristics as nearby areas with contaminated harbor sediment. The number of background stations and the number of sample replicates per background station depend on the statistical design of the sediment ecological risk investigation, developed through the data quality objective (DQO) process. Biological data from the background stations can be compared to data from a contaminated site by using minimum or maximum background levels or comparative statistics. In Navy ecological risk assessments (ERA's), calculated background levels and appropriate ecological risk screening criteria are used to identify sampling stations and sites with contaminated

  4. Cold Spring Harbor symposia on quantitative biology. Volume XLVII, Part 1. Structures of DNA

    International Nuclear Information System (INIS)

    Anon.

    1983-01-01

    The proceedings for the 47th Annual Cold Spring Harbor Symposia on Quantitative Biology are presented. This symposium focused on the Structure of DNA. Topics presented covered research in the handedness of DNA, conformational analysis, chemically modified DNA, chemical synthesis of DNA, DNA-protein interactions, DNA within nucleosomes, DNA methylation, DNA replication, gyrases and topoisomerases, recombining and mutating DNA, transcription of DNA and its regulation, the organization of genes along DNA, repetitive DNA and pseudogenes, and origins of replication, centromeres, and teleomeres

  5. The War on Cancer: Cold Spring Harbor Laboratory Is Fighting the Good Fight.

    Science.gov (United States)

    Mertz, Leslie

    2017-01-01

    Located on the north shore of Long Island in New York, Cold Spring Harbor Laboratory (Figure 1) started out with a marine biology emphasis at the end of the 19th century, but it soon established itself as a prominent cancer research facility. That strong emphasis on cancer work continues today as this private, not-for-profit research institution enters its 127th year (Figure 2).

  6. 33 CFR 165.150 - New Haven Harbor, Quinnipiac River, Mill River.

    Science.gov (United States)

    2010-07-01

    ... River, Mill River. 165.150 Section 165.150 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF... New Haven Harbor, Quinnipiac River, Mill River. (a) The following is a regulated navigation area: The... 303°T to point D at the west bank of the mouth of the Mill River 41°18′05″ N, 72°54′23″ W thence south...

  7. The Secant Rate of Corrosion: Correlating Observations of the USS Arizona Submerged in Pearl Harbor

    Science.gov (United States)

    Johnson, Donald L.; DeAngelis, Robert J.; Medlin, Dana J.; Johnson, Jon E.; Carr, James D.; Conlin, David L.

    2018-03-01

    Contrary to previous linear projections of steel corrosion in seawater, analysis of an inert marker embedded in USS Arizona concretion since the 7 December 1941 attack on Pearl Harbor reveals evidence that the effective corrosion rate decreases with time. The secant rate of corrosion, or SRC correlation, derived from this discovery could have a significant impact on failure analysis investigations for concreted shipwrecks or underwater structures. The correlation yields a lower rate of metal thinning than predicted. Development of the correlation is described.

  8. A Comprehensive Copper Compliance Strategy: Implementing Regulatory Guidance at Pearl Harbor Naval Shipyard & Intermediate Maintenance Facility

    Science.gov (United States)

    2007-01-01

    species was not listed in the PHLP database (Bishop Museum, 1998). Personal observations by John Zardus (Kewalo Marine Laboratory, Honolulu, Hawaii...Ringwood, 1992). Specimens collected from an open coastal site by the University of Hawaii ( John Zardus ), did not survive the shipment (less than 24... John Ornellas, Randy Kido, and Rich Anderson from Code 106.3. The Shipyard Pearl Harbor Regional Dive Locker, Code 760, provided professional, high

  9. Cold Spring Harbor symposia on quantitative biology: Volume 49, Recombination at the DNA level

    International Nuclear Information System (INIS)

    1984-01-01

    This volume contains full papers prepared by the participants to the 1984 Cold Springs Harbor Symposia on Quantitative Biology. This year's theme is entitled Recombination at the DNA level. The volume consists of 93 articles grouped into subject areas entitled chromosome mechanics, yeast systems, mammalian homologous recombination, transposons, mu, plant transposons/T4 recombination, topoisomerase, resolvase and gyrase, Escherichia coli general recombination, RecA, repair, leukaryotic enzymes, integration and excision of bacteriophage, site-specific recombination, and recombination in vitro

  10. The Plurality of Harbors at Caesarea: The Southern Anchorage in Late Antiquity

    Science.gov (United States)

    Ratzlaff, Alexandra; Galili, Ehud; Waiman-Barak, Paula; Yasur-Landau, Assaf

    2017-08-01

    The engineering marvel of Sebastos, or Portus Augusti as it was called in Late Antiquity (284-638 CE), dominated Caesarea's harbor center along modern Israel's central coast but it was only one part of a larger maritime complex. The Southern Anchorage provides a case study as one portion of the Caesarea complex, as well as a node within the regional network of anchorages and small harbors. Ceramics recovered from here show a high percentage of locally, and provincially, produced storage jars engaged in maritime trade. The ceramic evidence points towards an intensified regional trade or cabotage rather than favouring long distance trade from large port to port. Working out of these small harbors, opportunities arose for greater flexibility in specialization of commodities and materials passing through the network of subsidiary ports, contributing to a more diversified market economy. This analysis provides another example in the growing focus on how these simple and semi-modified anchorages in the Eastern Mediterranean were often the predominant economic networks connecting hinterland and coastal trade.

  11. Faunal and vegetation monitoring in response to harbor dredging in the Port of Miami

    Science.gov (United States)

    Daniels, Andre; Stevenson, Rachael; Smith, Erin; Robblee, Michael

    2018-04-11

    Seagrasses are highly productive ecosystems. A before-after-control-impact (BACI) design was used to examine effects of dredging on seagrasses and the animals that inhabit them. The control site North Biscayne Bay and the affected site Port of Miami had seagrass densities decrease during both the before, Fish and Invertebrate Assessment Network 2006-2011, and after, Faunal Monitoring in Response to Harbor Dredging 2014-2016, studies. Turbidity levels increased at North Biscayne Bay and Port of Miami basins during the Faunal Monitoring in Response to Harbor Dredging study, especially in 2016. Animal populations decreased significantly in North Biscayne Bay and Port of Miami in the Faunal Monitoring in Response to Harbor Dredging study compared to the Fish and Invertebrate Assessment Network study. Predictive modeling shows that numbers of animal populations will likely continue to decrease if the negative trends in seagrass densities continue unabated. There could be effects on several fisheries vital to the south Florida economy. Additional research could determine if animal populations and seagrass densities have rebounded or continued to decrease.

  12. Cloning and Sequencing of Protein Kinase cDNA from Harbor Seal (Phoca vitulina Lymphocytes

    Directory of Open Access Journals (Sweden)

    Jennifer C. C. Neale

    2004-01-01

    Full Text Available Protein kinases (PKs play critical roles in signal transduction and activation of lymphocytes. The identification of PK genes provides a tool for understanding mechanisms of immunotoxic xenobiotics. As part of a larger study investigating persistent organic pollutants in the harbor seal and their possible immunomodulatory actions, we sequenced harbor seal cDNA fragments encoding PKs. The procedure, using degenerate primers based on conserved motifs of human protein tyrosine kinases (PTKs, successfully amplified nine phocid PK gene fragments with high homology to human and rodent orthologs. We identified eight PTKs and one dual (serine/threonine and tyrosine kinase. Among these were several PKs important in early signaling events through the B- and T-cell receptors (FYN, LYN, ITK and SYK and a MAP kinase involved in downstream signal transduction. V-FGR, RET and DDR2 were also expressed. Sequential activation of protein kinases ultimately induces gene transcription leading to the proliferation and differentiation of lymphocytes critical to adaptive immunity. PKs are potential targets of bioactive xenobiotics, including persistent organic pollutants of the marine environment; characterization of these molecules in the harbor seal provides a foundation for further research illuminating mechanisms of action of contaminants speculated to contribute to large-scale die-offs of marine mammals via immunosuppression.

  13. Length-weight relationship of northern pike, Esox lucius, from East Harbor, Ohio

    Science.gov (United States)

    Brown, Edward H.; Clark, Clarence F.

    1965-01-01

    The northern pike is one of Ohio's largest game fish but is well known to comparatively few anglers. Large numbers of the big fish spawn in the Ohio marshes adjacent to Lake Erie. Movements related to spawning reach a peak in late March or early April. Later the spawning population disperses and is seldom represented in catches by experimental gear or by anglers. The short period of availability was used to obtain life history information in March of 1951 through 1953. No comprehensive length-weight data for this species have previously been published from this area. East Harbor is a sandspit pond separated from Lake Erie by a large sand bar. Waters and fish populations of the harbor and lake can mix freely through a permanent connecting channel. The larger part of the 850 surface acres of the harbor is normally less than 8 feet deep. The male northern pike averaged 20.5 inches in length and ranged from 13.5 to 28.5 inches. The conspicuously larger females averaged 26.0 inches and ranged from 15.5 to 37.5 inches.

  14. Composition and source of butyltins in sediments of Kaohsiung Harbor, Taiwan

    Science.gov (United States)

    Dong, Cheng-Di; Chen, Chih-Feng; Chen, Chiu-Wen

    2015-04-01

    Fifty-eight sediment samples were collected from the Kaohsiung Harbor (Taiwan) for analyses of monobutyltin (MBT), dibutyltin (DBT) and tributyltin (TBT), using gas chromatography/flame photometric detector (GC/FPD). The concentration of total butyltins (ΣBTs), sum of MBT, DBT, and TBT, varied from 3.9 to 158.5 ng Sn/g dw in sediment samples with TBT being the major component of the sediment samples, except for the vicinity of the Love River mouth where MBT was the most abundant BT compound (a proportion of over 57%). Based on the BTs concentration, distribution, composition and correlations, the sources of BTs found in harbor sediments are shipping activities, and TBT is the main pollutant; the estuary (i.e. Love River) has been the anthropogenic source of MBT from upstream inputs. Influences of TBT on aquatic organisms are evaluated using the toxicity guidelines proposed by the US EPA (US Environmental Protection Agency) and the ACCI (assessment class criterion for imposex) proposed by OSPAR (Oslo and Paris Commission). The evaluation shows that the TBT contained in the sediment at Kaohsiung Harbor is likely to have a negative influence at ACCI class C because gastropods present imposex and TBT levels are above ecotoxicological assessment criteria (EAC) limits.

  15. A Comparison of Microbial Water Quality and Diversity for Ballast and Tropical Harbor Waters.

    Directory of Open Access Journals (Sweden)

    Charmaine Ng

    Full Text Available Indicator organisms and antibiotic resistance were used as a proxy to measure microbial water quality of ballast tanks of ships, and surface waters in a tropical harbor. The survival of marine bacteria in ballast tanks appeared to diminish over longer water retention time, with a reduction of cell viability observed after a week based on heterotrophic plate counts. Pyrosequencing of 16S rRNA genes showed distinct differences in microbial composition of ballast and harbor waters. The harbor waters had a higher abundance of operational taxonomic units (OTUs assigned to Cyanobacteria (Synechococcus spp. and α-proteobacteria (SAR11 members, while marine hydrocarbon degraders such as γ-proteobacteria (Ocenspirillaes spp., Thiotrchales spp. and Bacteroidetes (Flavobacteriales spp. dominated the ballast water samples. Screening of indicator organisms found Escherichia coli (E. coli, Enterococcus and Pseudomonas aeruginosa (P. aeruginosa in two or more of the ballast and harbor water samples tested. Vibrio spp. and Salmonella spp. were detected exclusively in harbor water samples. Using quantitative PCR (qPCR, we screened for 13 antibiotic resistant gene (ARG targets and found higher abundances of sul1 (4.13-3.44 x 102 copies/mL, dfrA (0.77-1.80 x10 copies/mL and cfr (2.00-5.21 copies/mL genes compared to the other ARG targets selected for this survey. These genes encode for resistance to sulfonamides, trimethoprim and chloramphenicol-florfenicol antibiotics, which are also known to persist in sediments of aquaculture farms and coastal environments. Among the ARGs screened, we found significant correlations (P<0.05 between ereA, ermG, cfr and tetO genes to one or more of the indicator organisms detected in this study, which may suggest that these members contribute to the environmental resistome. This study provides a baseline water quality survey, quantitatively assessing indicators of antibiotic resistance, potentially pathogenic organisms and a

  16. Selenium and mercury concentrations in harbor seals (Phoca vitulina) from central California: Health implications in an urbanized estuary

    Science.gov (United States)

    McHuron, Elizabeth A.; Harvey, James T.; Castellini, J. Margaret; Stricker, Craig A.; O'Hara, Todd M.

    2014-01-01

    We measured total selenium and total mercury concentrations ([TSe] and [THg]) in hair (n = 138) and blood (n = 73) of harbor seals (Phoca vitulina) from California to assess variation by geography and sex, and inferred feeding relationships based on carbon, nitrogen, and sulfur stable isotopes. Harbor seals from Hg-contaminated sites had significantly greater [THg], and lesser [TSe] and TSe:THg molar ratios than seals from a relatively uncontaminated site. Males had significantly greater [THg] than females at all locations. Sulfur stable isotope values explained approximately 25% of the variability in [THg], indicating increased Hg exposure for seals with a greater use of estuarine prey species. Decreased [TSe] in harbor seals from Hg-contaminated regions may indicate a relative Se deficiency to mitigate the toxic effects of Hg. Further investigation into the Se status and the potential negative impact of Hg on harbor seals from Hg-contaminated sites is warranted.

  17. Comparative account of macrofouling ecology at Vijaydurg Harbor and Musakazi-Jaitapur Bay, central western coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Sawant, S.S.; Wagh, A.B.

    The extent and the nature of fouling were studied on mild steel, aluminum, and stainless steel panels at the Vijaydurg Harbor and Musakazi-Jaitapur Bay on the central western coast of India. In addition, cathodically protected and unprotected panels...

  18. Biodiversity of marine communities in Pearl Harbor, Oahu, Hawaii with observations in 1996 on introduced exotic species (NODC Accession 0000330)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The marine and estuarine invertebrate and fish communities in Pearl Harbor, Oahu, Hawaii were surveyed between January and October, 1996. Samples were taken and...

  19. Benthic grab data from October 1994 in the Lower Bay of New York/New Jersey Harbor

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Benthic grab samples were collected using a Shipek grab at pre-determined locations in the harbor. Taxonomic enumerations and biological community parameters were...

  20. Directional wave and temperature data from nine buoys in Gray's Harbor, Washington, 1994-2002 (NODC Accession 0000756)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Wave data were collected from 9 buoys in Grays Harbor, Washington, from 01 January 1994 to 24 July 2002. Data were collected as part of the Coastal Data Information...

  1. Central nervous system disease and genital disease in harbor porpoises (Phocoena phocoena) are associated with different herpesviruses

    NARCIS (Netherlands)

    C.E. van Elk; M.W.G. van de Bildt (Marco); P.R.W.A. van Run (Peter); De Jong, A. (Anton); S. Getu (Sarah); G.M.G.M. Verjans (George); A.D.M.E. Osterhaus (Albert); T. Kuiken (Thijs)

    2016-01-01

    textabstractHerpesvirus infection causes disease of variable severity in many species, including cetaceans. However, little is known about herpesvirus infection in harbor porpoises (Phocoena phocoena), despite being widespread in temperate coastal waters of the Northern Hemisphere. Therefore, we

  2. Case study of small harbor excitation under storm and tsunami conditions

    Science.gov (United States)

    Synolakis, Costas; Maravelakis, Nikos; Kalligeris, Nikos; Skanavis, Vassilios; Kanoglu, Utku; Yalciner, Ahmet; Lynett, Pat

    2016-04-01

    Simultaneous nearshore and interior-to-ports wave and current measurements for small ports are not common, and few, if any, benchmarking cases at sufficient resolution exist to help validate numerical model of intermediate waves, or even long waves. The wave conditions inside the old Venetian harbor of Chania, Greece and offshore were measured and studied from 2012 to 2015. The construction of this harbor began in the 14th century, and since then, its layout has been modified to adapt to different social and to economic conditions. It is divided into a western and an eastern basin. The eastern basin is used by recreational vessels and fishing boats throughout the year. The western basin has an exposed entrance to the north, and it is essentially functional half of the year, because of the severe overtopping and flooding that occur during the northern winter storms. Our work is motivated by the necessity to protect the monument from severe winter storm conditions and allow safe mooring and all other recreational activities that take place in the exposed western basin. Two earlier studies had proposed the construction of a low crested breakwater near the harbor entrance. The first design has been partially constructed, while the second never materialized. The main disadvantage of both studies was the lack of any wave field measurements. At the same time, second order or complimentary phenomena such as harbor resonance had not been considered. To address the lack of field data, the offshore wave climate has been monitored since October 2012 using an AWAC 600kHz instrument, deployed at 23m depth. The response of the western and eastern basins of the harbor was measured with a TWR-2050 (deployed at 5.5m depth) and an RBRDuet T.D./wave (deployed at 2m depth) pressure gauges respectively. Significant wave heights ranging up to 5.8 m with significant periods of up to 10 sec were measured. The harbor pressure gauges are now being re-deployed in other locations to collect

  3. Sediment quality in Burlington Harbor, Lake Champlain, U.S.A.

    Science.gov (United States)

    Lacey, E.M.; King, J.W.; Quinn, J.G.; Mecray, E.L.; Appleby, P.G.; Hunt, A.S.

    2001-01-01

    Surface samples and cores were collected in 1993 from the Burlington Harbor region of Lake Champlain. Sediment samples were analyzed for trace metals (cadmium, copper, lead, nickel, silver and zinc), simultaneously extracted metal/acid volatile sulfide (SEM-AVS), grain size, nutrients (carbon and nitrogen) and organic contaminants (polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs)). The concentrations of cadmium, copper, silver and zinc from the partial sediment digestion of the surface samples correlated well with each other (r2 > 0.60) indicating that either a common process, or group of processes determined the sediment concentrations of these metals. In an analysis of the spatial distribution of the trace metals and PAHs, high surficial concentrations were present in the southern portion of the Harbor. The trace metal trend was strengthened when the concentrations were normalized by grain size. A sewage treatment plant outfall discharge was present in the southeastern portion of the Harbor at the time of this study and is the major source of trace metal and PAH contamination. Evaluation of sediment cores provides a proxy record of historical trace metal and organic inputs. The peak accumulation rate for copper, cadmium, lead, and zinc was in the late 1960s and the peak silver accumulation rate was later. The greatest accumulation of trace metals occurred in the late 1960s after discharges from the STP began. Subsequent declines in trace metal concentrations may be attributed to increased water and air regulations. The potential toxicity of trace metals and organic contaminants was predicted by comparing contaminant concentrations to benchmark concentrations and potential trace metal bioavailability was predicted with SEM-AVS results. Surface sample results indicate lead, silver, ???PAHs and ???PCBs are potentially toxic and/or bioavailable. These predictions were supported by studies of biota in the Burlington Harbor watershed. There

  4. Benthic foraminifera distribution in high polluted sediments from Niterói Harbor (Guanabara Bay), Rio de Janeiro, Brazil

    OpenAIRE

    Claudia G. Vilela; Daniele S. Batista; José A. Batista-Neto; Mirian Crapez; John J. Mcallister

    2004-01-01

    Dockyards and harbors are recognized as being important locations where sediment-associated pollutants can accumulate, which constitutes an environmental risk to aquatic life due to potential uptake and accumulation of heavy metals in the biota. The aim of this paper is to assess the concentrations and the effects of some heavy metals in the benthic foraminifera assemblage in Niterói Harbor. Low concentrations in the benthic foraminifera as well as the dominance of indicative species such as ...

  5. Development of an androgen reporter gene assay (AR-LUX) utilizing a human cell line with an endogenously regulated androgen receptor

    NARCIS (Netherlands)

    Blankvoort, B.M.G.; Groene, E.M. de; Meeteren-Kreikamp, A.P. van; Witkamp, R.F.; Rodenburg, R.J.T.; Aarts, J.M.M.J.G.

    2001-01-01

    The aim of the work described in this report is to develop and characterize a cell-based androgen reporter assay. For this purpose, the androgen receptor (AR) expressing human breast cancer cell line T47D was stably transfected with a luciferase gene under transcriptional control of the PB-ARE-2

  6. New genomic resources for switchgrass: a BAC library and comparative analysis of homoeologous genomic regions harboring bioenergy traits

    Directory of Open Access Journals (Sweden)

    Feltus Frank A

    2011-07-01

    Full Text Available Abstract Background Switchgrass, a C4 species and a warm-season grass native to the prairies of North America, has been targeted for development into an herbaceous biomass fuel crop. Genetic improvement of switchgrass feedstock traits through marker-assisted breeding and biotechnology approaches calls for genomic tools development. Establishment of integrated physical and genetic maps for switchgrass will accelerate mapping of value added traits useful to breeding programs and to isolate important target genes using map based cloning. The reported polyploidy series in switchgrass ranges from diploid (2X = 18 to duodecaploid (12X = 108. Like in other large, repeat-rich plant genomes, this genomic complexity will hinder whole genome sequencing efforts. An extensive physical map providing enough information to resolve the homoeologous genomes would provide the necessary framework for accurate assembly of the switchgrass genome. Results A switchgrass BAC library constructed by partial digestion of nuclear DNA with EcoRI contains 147,456 clones covering the effective genome approximately 10 times based on a genome size of 3.2 Gigabases (~1.6 Gb effective. Restriction digestion and PFGE analysis of 234 randomly chosen BACs indicated that 95% of the clones contained inserts, ranging from 60 to 180 kb with an average of 120 kb. Comparative sequence analysis of two homoeologous genomic regions harboring orthologs of the rice OsBRI1 locus, a low-copy gene encoding a putative protein kinase and associated with biomass, revealed that orthologous clones from homoeologous chromosomes can be unambiguously distinguished from each other and correctly assembled to respective fingerprint contigs. Thus, the data obtained not only provide genomic resources for further analysis of switchgrass genome, but also improve efforts for an accurate genome sequencing strategy. Conclusions The construction of the first switchgrass BAC library and comparative analysis of

  7. A metastatic colon adenocarcinoma harboring BRAF V600E has a durable major response to dabrafenib/trametinib and chemotherapy

    Directory of Open Access Journals (Sweden)

    Williams CB

    2015-12-01

    Full Text Available Casey B Williams,1,* Caitlin McMahon,2,* Siraj M Ali,2 Mark Abramovitz,1 Kirstin A Williams,1 Jessica Klein,1 Heidi McKean,1 Roman Yelensky,2 Thomas J George Jr,3 Julia A Elvin,2 Salil Soman,4 Doron Lipson,2 Juliann Chmielecki,2 Deborah Morosini,2 Vincent A Miller,2 Philip J Stephens,2 Jeffrey S Ross,2,5 Brian Leyland-Jones1 1Avera Cancer Institute, Sioux Falls, SD, USA; 2Foundation Medicine, Inc., Cambridge, MA, USA; 3University of Florida College of Medicine, Gainesville, FL, USA; 4Beth Israel Deaconess Medical Center, Boston, MA, USA; 5Albany Medical College, Albany, NY, USA *These authors contributed equally to this work Abstract: The subset of metastatic colorectal adenocarcinomas that harbor BRAF V600E mutations are aggressive tumors with significantly shortened survival and limited treatment options. Here we present a colorectal cancer patient whose disease progressed through standard chemotherapy and who developed liver metastasis. Comprehensive genomic profiling (FoundationOne® identified a BRAF V600E mutation in the liver lesion, as well as other genomic alterations consistent with colorectal cancers. Combination therapy of dabrafenib and trametinib with standard cytotoxic chemotherapy resulted in a durable major ongoing response for the patient. This report illustrates the utility of comprehensive genomic profiling with personalized targeted therapy for aggressive metastatic colorectal adenocarcinomas. Keywords: oxaliplatin, colorectal adenocarcinoma, combination targeted therapy, BRAF mutations

  8. Persistence of Multidrug-Resistant Acinetobacter baumannii Isolates Harboring blaOXA-23 and bap for 5 Years.

    Science.gov (United States)

    Sung, Ji Youn; Koo, Sun Hoe; Kim, Semi; Kwon, Gye Cheol

    2016-08-28

    The emergence and dissemination of carbapenemase-producing Acinetobacter baumannii isolates have been reported worldwide, and A. baumannii isolates harboring blaOXA-23 are often resistant to various antimicrobial agents. Antimicrobial resistance can be particularly strong for biofilm-forming A. baumannii isolates. We investigated the genetic basis for carbapenem resistance and biofilm-forming ability of multidrug-resistant (MDR) clinical isolates. Ninety-two MDR A. baumannii isolates were collected from one university hospital located in the Chungcheong area of Korea over a 5-year period. Multiplex PCR and DNA sequencing were performed to characterize carbapenemase and bap genes. Clonal characteristics were analyzed using REP-PCR. In addition, imaging and quantification of biofilms were performed using a crystal violet assay. All 92 MDR A. baumannii isolates involved in our study contained the blaOXA-23 and bap genes. The average absorbance of biomass in Bap-producing strains was much greater than that in non-Bap-producing strains. In our study, only three REP-PCR types were found, and the isolates showing type A or type B were found more than 60 times among unique patients during the 5 years of surveillance. These results suggest that the isolates have persisted and colonized for 5 years, and biofilm formation ability has been responsible for their persistence and colonization.

  9. An ATP synthase harboring an atypical γ-subunit is involved in ATP synthesis in tomato fruit chromoplasts.

    Science.gov (United States)

    Pateraki, Irini; Renato, Marta; Azcón-Bieto, Joaquín; Boronat, Albert

    2013-04-01

    Chromoplasts are non-photosynthetic plastids specialized in the synthesis and accumulation of carotenoids. During fruit ripening, chloroplasts differentiate into photosynthetically inactive chromoplasts in a process characterized by the degradation of the thylakoid membranes, and by the active synthesis and accumulation of carotenoids. This transition renders chromoplasts unable to photochemically synthesize ATP, and therefore these organelles need to obtain the ATP required for anabolic processes through alternative sources. It is widely accepted that the ATP used for biosynthetic processes in non-photosynthetic plastids is imported from the cytosol or is obtained through glycolysis. In this work, however, we show that isolated tomato (Solanum lycopersicum) fruit chromoplasts are able to synthesize ATP de novo through a respiratory pathway using NADPH as an electron donor. We also report the involvement of a plastidial ATP synthase harboring an atypical γ-subunit induced during ripening, which lacks the regulatory dithiol domain present in plant and algae chloroplast γ-subunits. Silencing of this atypical γ-subunit during fruit ripening impairs the capacity of isolated chromoplast to synthesize ATP de novo. We propose that the replacement of the γ-subunit present in tomato leaf and green fruit chloroplasts by the atypical γ-subunit lacking the dithiol domain during fruit ripening reflects evolutionary changes, which allow the operation of chromoplast ATP synthase under the particular physiological conditions found in this organelle. © 2013 The Authors The Plant Journal © 2013 Blackwell Publishing Ltd.

  10. Yeast transformation mediated by Agrobacterium strains harboring an Ri plasmid: comparative study between GALLS of an Ri plasmid and virE of a Ti plasmid.

    Science.gov (United States)

    Kiyokawa, Kazuya; Yamamoto, Shinji; Sato, Yukari; Momota, Naoto; Tanaka, Katsuyuki; Moriguchi, Kazuki; Suzuki, Katsunori

    2012-07-01

    Agrobacterium strains containing a Ti plasmid can transfer T-DNA not only to plants but also to fungi, including the yeast Saccharomyces cerevisiae. However, no Agrobacterium strain harboring an Ri plasmid has been evaluated in fungal transformation. Some Ri plasmids have GALLS , instead of virE1 and virE2. GALLS protein can functionally substitute in plant transformation for a structurally different protein VirE2. In this study, we compared the yeast transformation ability among Agrobacterium donors: a strain containing a Ti plasmid, strains harboring either an agropine-type or a mikimopine-type Ri plasmid, and a strain having a modified Ri plasmid supplemented with a Ti plasmid type virE operon. Agrobacterium strains possessing GALLS transformed yeast cells far less efficiently than the strain containing virE operon. Production of GALLS in recipient yeast cells improved the yeast transformation mediated by an Agrobacterium strain lacking neither GALLS nor virE operon. A reporter assay to detect mobilization of the proteins fused with Cre recombinase revealed that VirE2 protein is much more abundant in yeast cells than GALLS. Based on these results, we concluded that the low yeast transformability mediated by Agrobacterium strains having the Ri plasmid is because of low amount of mobilized GALLS in yeast cells. © 2012 The Authors Journal compilation © 2012 by the Molecular Biology Society of Japan/Blackwell Publishing Ltd.

  11. Annexin A2 promotes the migration and invasion of human hepatocellular carcinoma cells in vitro by regulating the shedding of CD147-harboring microvesicles from tumor cells.

    Directory of Open Access Journals (Sweden)

    Wei Zhang

    Full Text Available It has been reported that Annexin A2 (ANXA2 is up-regulated in hepatocellular carcinoma (HCC, but the roles of ANXA2 in the migration and invasion of HCC cells have not been determined. In this study, we found that ANXA2-specific siRNA (si-ANXA2 significantly inhibited the migration and invasion of HCC cells co-cultured with fibroblasts in vitro. In addition, the production of MMP-2 by fibroblasts cultured in supernatant collected from si-ANXA2-transfected HCC cells was notably down-regulated. ANXA2 was also found to be co-localized and co-immunoprecipitated with CD147. Further investigation revealed that the expression of ANXA2 in HCC cells affected the shedding of CD147-harboring membrane microvesicles, acting as a vehicle for CD147 in tumor-stromal interactions and thereby regulating the production of MMP-2 by fibroblasts. Together, these results suggest that ANXA2 enhances the migration and invasion potential of HCC cells in vitro by regulating the trafficking of CD147-harboring membrane microvesicles.

  12. A Carbapenem-Resistant Pseudomonas aeruginosa Isolate Harboring Two Copies of blaIMP-34 Encoding a Metallo-β-Lactamase.

    Directory of Open Access Journals (Sweden)

    Tatsuya Tada

    Full Text Available A carbapenem-resistant strain of Pseudomonas aeruginosa, NCGM1984, was isolated in 2012 from a hospitalized patient in Japan. Immunochromatographic assay showed that the isolate was positive for IMP-type metallo-β-lactamase. Complete genome sequencing revealed that NCGM1984 harbored two copies of blaIMP-34, located at different sites on the chromosome. Each blaIMP-34 was present in the same structures of the class 1 integrons, tnpA(ISPa7-intI1-qacG-blaIMP-34-aac(6'-Ib-qacEdelta1-sul1-orf5-tniBdelta-tniA. The isolate belonged to multilocus sequence typing ST235, one of the international high-risk clones. IMP-34, with an amino acid substitution (Glu126Gly compared with IMP-1, hydrolyzed all β-lactamases tested except aztreonam, and its catalytic activities were similar to IMP-1. This is the first report of a clinical isolate of an IMP-34-producing P. aeruginosa harboring two copies of blaIMP-34 on its chromosome.

  13. Description and proposed life cycle of Maritrema novaezealandensis n. sp. (Microphallidae) parasitic in red-billed gulls, Larus novaehollandiae scopulinus, from Otago Harbor, South Island, New Zealand.

    Science.gov (United States)

    Martorelli, Sergio R; Fredensborg, Brian L; Mouritsen, Kim N; Poulin, Robert

    2004-04-01

    Maritrema novaezealandensis n. sp. is described from Otago Harbor, South Island, New Zealand, on the basis of adult specimens collected from the Red-billed gull, Larus novaehollandiae scopulinus, and excysted metacercariae obtained from crabs. It belongs to the "eroliae group" and differs from other related species mainly in the shape, size, and patterns of distributions of the spines on the cirrus, the shape of the metraterm, the presence of an unlobed ovary, and the complete ring of the vitelline follicles. Based on morphometric features of metacercariae and adult specimens, the trophic relationships among invertebrate and vertebrate hosts, experimental infections, and previous reports of species of Maritrema with similar transmission patterns, the life cycle of M. novaezealandensis n. sp. is described. A 3-host life cycle is proposed for this parasite. The first intermediate host is the mud snail, Zeacumantus subcarinatus, in which the cercarial stage is produced in sporocysts located within the gonad of the snail. At least 3 crab species (Hemigrapsus crenulatus, Macrophtalmus hirtipes, and Halicarcinus whitei) and several species of amphipods act as second intermediate hosts, with metacercariae encysted in the body cavity of the crustacean host. Finally, the definitive host, the gull, L. n. scopulinus, harbors the adult worms in its intestine.

  14. The examination of urine samples for pathogenic microbes by the luciferase assay for ATP. 1: The effect of the presence of fungi, fungal like bacteria and kidney cells in urine samples

    Science.gov (United States)

    Bush, V. N.

    1973-01-01

    A method for accurately determining urinary tract infections in man is introduced. The method is based on adenosine triphosphate (ATP) concentration in urine samples after removing nonbacterial ATP. Adenosine triphosphate concentration is measured from the bioluminescent reaction of luciferase when mixed with ATP. An examination was also made of the effectiveness of rupturing agents on monkey kidney cells Candia albicans, a Rhodotorula species, and a Streptomyces species in determining whether these cells could contribute ATP to the bacterial ATP value of a urine sample.

  15. Assessment of the impact of oil spill on Mumbai harbor bay

    International Nuclear Information System (INIS)

    Pandit, G.G.; Saradhi, I.V.; Puranik, V.D.; Sharma, D.N.; Ghosh, A.K.

    2011-04-01

    On 7 th August, 2010 two cargo ships MSC Chitra and MV Khalijia collided off Mumbai coast causing an oil spill that spread quickly through Maharashtra's coastline. MSC Chitra ruptured its tank when it hit incoming MV Khalijia. MSC Chitra tilted to about 80 degrees soon after the collision, spilling an estimated 400 to 500 tonnes of oil. The ship was loaded with an estimated 2,600 tonnes of oil, 300 tonnes of diesel and 70 tonnes of lubricating oil at the time of the accident. The oil spill has spreaded over an area of 25 square kilometers. MSC Chitra was carrying 1219 containers out of which 31 had hazardous chemicals like organophosphate pesticides. After the collision about 400 containers fall off into to the sea from the deck of MSC Chitra. The oil spill has resulted in severe environmental damage along the Mumbai coastline as well as to the marine life. In order to assess the impact of oil spill on the marine environment daily monitoring of oil and grease levels has been carried out at CIRUS Jetty. Additionally three field surveys have been carried out along the coast line of Mumbai harbor bay from Vashi Jetty to Uran collecting sea water samples. The samples collected at CIRUS Jetty and Mumbai harbor bay are analyzed for various water quality parameters apart from oil and grease. The results indicate the oil and grease levels in sea water at CIRUS Jetty reduced to background levels within 15 days. The filed survey along Mumbai harbor bay also shows a decreasing trend in the oil and grease levels in seawater. (author)

  16. Clinical and CT characteristics of surgically resected lung adenocarcinomas harboring ALK rearrangements or EGFR mutations

    International Nuclear Information System (INIS)

    Wang, Hua; Schabath, Matthew B.; Liu, Ying; Han, Ying; Li, Qi; Gillies, Robert J.; Ye, Zhaoxiang

    2016-01-01

    Purpose: To determine if clinical and CT characteristics of surgically resected lung adenocarcinomas can distinguish those harboring ALK rearrangements from EGFR mutations. Materials and methods: Patients who had surgical resection and histologically confirmed lung adenocarcinoma were enrolled, including 41 patients with ALK rearrangements and 66 patients with EGFR mutations. Eighteen categorical and six quantitative CT characteristics were used to evaluate the tumors. Differences in clinical and CT characteristics between the two groups were investigated. Results: Age (P = 0.003), histological subtypes (P < 0.001), pathological stage (P = 0.007), and five CT characteristics, including size (P < 0.001), GGO (P = 0.001), bubble-like lucency (P = 0.048), lymphadenopathy (P = 0.001), and tumor shadow disappearance rate (P = 0.005) were significantly different between patients harboring ALK rearrangements compared to patients with EGFR mutations. When we compared histologic components, a solid pattern was more common (P = 0.009) in tumors with ALK rearrangements, and lepidic and acinar patterns were more common (P < 0.001 and P = 0.040, respectively) in those with EGFR mutations. Backward elimination analyses revealed that age (OR = 0.93; 95% CI 0.89–0.98), GGO (OR = 0.14; 95% CI 0.03–0.67), and lymphadenopathy (OR = 4.15; 95% CI 1.49–11.60) were significantly associated with ALK rearrangement status. Conclusion: Our analyses revealed that clinical and CT characteristics of lung adenocarcinomas harboring ALK rearrangements were significantly different, compared with those with EGFR mutations. These differences may be related to the molecular pathology of these diseases.

  17. Influenza A (H10N7 Virus Causes Respiratory Tract Disease in Harbor Seals and Ferrets.

    Directory of Open Access Journals (Sweden)

    Judith M A van den Brand

    Full Text Available Avian influenza viruses sporadically cross the species barrier to mammals, including humans, in which they may cause epidemic disease. Recently such an epidemic occurred due to the emergence of avian influenza virus of the subtype H10N7 (Seal/H10N7 in harbor seals (Phoca vitulina. This epidemic caused high mortality in seals along the north-west coast of Europe and represented a potential risk for human health. To characterize the spectrum of lesions and to identify the target cells and viral distribution, findings in 16 harbor seals spontaneously infected with Seal/H10N7 are described. The seals had respiratory tract inflammation extending from the nasal cavity to bronchi associated with intralesional virus antigen in respiratory epithelial cells. Virus infection was restricted to the respiratory tract. The fatal outcome of the viral infection in seals was most likely caused by secondary bacterial infections. To investigate the pathogenic potential of H10N7 infection for humans, we inoculated the seal virus intratracheally into six ferrets and performed pathological and virological analyses at 3 and 7 days post inoculation. These experimentally inoculated ferrets displayed mild clinical signs, virus excretion from the pharynx and respiratory tract inflammation extending from bronchi to alveoli that was associated with virus antigen expression exclusively in the respiratory epithelium. Virus was isolated only from the respiratory tract. In conclusion, Seal/H10N7 infection in naturally infected harbor seals and experimentally infected ferrets shows that respiratory epithelial cells are the permissive cells for viral replication. Fatal outcome in seals was caused by secondary bacterial pneumonia similar to that in fatal human cases during influenza pandemics. Productive infection of ferrets indicates that seal/H10N7 may possess a zoonotic potential. This outbreak of LPAI from wild birds to seals demonstrates the risk of such occasions for mammals

  18. NifH- Harboring Bacterial Community Composition Across an Alaskan Permafrost Thaw Gradient

    Directory of Open Access Journals (Sweden)

    Christopher Ryan Penton

    2016-11-01

    Full Text Available Since nitrogen (N is often limiting in permafrost soils, we investigated the N2-fixing genetic potential and the inferred taxa harboring those genes by sequencing nifH gene fragments in samples taken along a permafrost thaw gradient in an Alaskan boreal soil. Samples from minimally, moderately and extensively thawed sites were taken to a depth of 79 cm to encompass zones above and below the depth of the water table. NifH reads were translated with frameshift correction and 112,476 sequences were clustered at 5% amino acid dissimilarity resulting in 1,631 OTUs. Sample depth in relation to water table depth was correlated to differences in the NifH sequence classes. NifH sequences most closely related to group I nifH-harboring Alpha- and Beta Proteobacteria were in higher abundance above water table depth while those related to group III nifH-harboring Delta and Gamma Proteobacteria were more abundant below. The most dominant below water table depth NifH sequences, comprising 1/3 of the total, were distantly related to Verrucomicrobia-Opitutaceae. Overall, these results suggest that permafrost thaw alters the class-level composition of N2-fixing communities in the thawed soil layers and that this distinction corresponds to the depth of the water table. These nifH data were also compared to nifH sequences obtained from a study at an Alaskan taiga site, and to those of other geographically distant, non-permafrost sites. The two Alaska sites were differentiated largely by changes in relative abundances of the same OTUs, whereas the non-Alaska sites were differentiated by the lack of many Alaskan OTUs, and the presence of unique halophilic, sulfate- and iron-reducing taxa in the Alaska sites.

  19. Characterization of differential gene expression in adrenocortical tumors harboring beta-catenin (CTNNB1) mutations.

    Science.gov (United States)

    Durand, Julien; Lampron, Antoine; Mazzuco, Tania L; Chapman, Audrey; Bourdeau, Isabelle

    2011-07-01

    Mutations of β-catenin gene (CTNNB1) are frequent in adrenocortical adenomas (AA) and adrenocortical carcinomas (ACC). However, the target genes of β-catenin have not yet been identified in adrenocortical tumors. Our objective was to identify genes deregulated in adrenocortical tumors harboring CTNNB1 genetic alterations and nuclear accumulation of β-catenin. Microarray analysis identified a dataset of genes that were differently expressed between AA with CTNNB1 mutations and wild-type (WT) tumors. Within this dataset, the expression profiles of five genes were validated by real time-PCR (RT-PCR) in a cohort of 34 adrenocortical tissues (six AA and one ACC with CTNNB1 mutations, 13 AA and four ACC with WT CTNNB1, and 10 normal adrenal glands) and two human ACC cell lines. We then studied the effects of suppressing β-catenin transcriptional activity with the T-cell factor/β-catenin inhibitors PKF115-584 and PNU74654 on gene expression in H295R and SW13 cells. RT-PCR analysis confirmed the overexpression of ISM1, RALBP1, and PDE2A and the down-regulation of PHYHIP in five of six AA harboring CTNNB1 mutations compared with WT AA (n = 13) and normal adrenal glands (n = 10). RALBP1 and PDE2A overexpression was also confirmed at the protein level by Western blotting analysis in mutated tumors. ENC1 was specifically overexpressed in three of three AA harboring CTNNB1 point mutations. mRNA expression and protein levels of RALBP1, PDE2A, and ENC1 were decreased in a dose-dependent manner in H295R cells after treatment with PKF115-584 or PNU74654. This study identified candidate genes deregulated in CTNNB1-mutated adrenocortical tumors that may lead to a better understanding of the role of the Wnt-β-catenin pathway in adrenocortical tumorigenesis.

  20. Clinical and CT characteristics of surgically resected lung adenocarcinomas harboring ALK rearrangements or EGFR mutations

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Hua [Department of Radiology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin (China); Schabath, Matthew B. [Department of Cancer Epidemiology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL (United States); Liu, Ying [Department of Radiology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin (China); Department of Cancer Imaging and Metabolism, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL (United States); Han, Ying [Department of Biotherapy, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin (China); Li, Qi [Department of Pathology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin (China); Gillies, Robert J. [Department of Cancer Imaging and Metabolism, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL (United States); Department of Radiology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL (United States); Ye, Zhaoxiang, E-mail: yezhaoxiang@163.com [Department of Radiology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin (China)

    2016-11-15

    Purpose: To determine if clinical and CT characteristics of surgically resected lung adenocarcinomas can distinguish those harboring ALK rearrangements from EGFR mutations. Materials and methods: Patients who had surgical resection and histologically confirmed lung adenocarcinoma were enrolled, including 41 patients with ALK rearrangements and 66 patients with EGFR mutations. Eighteen categorical and six quantitative CT characteristics were used to evaluate the tumors. Differences in clinical and CT characteristics between the two groups were investigated. Results: Age (P = 0.003), histological subtypes (P < 0.001), pathological stage (P = 0.007), and five CT characteristics, including size (P < 0.001), GGO (P = 0.001), bubble-like lucency (P = 0.048), lymphadenopathy (P = 0.001), and tumor shadow disappearance rate (P = 0.005) were significantly different between patients harboring ALK rearrangements compared to patients with EGFR mutations. When we compared histologic components, a solid pattern was more common (P = 0.009) in tumors with ALK rearrangements, and lepidic and acinar patterns were more common (P < 0.001 and P = 0.040, respectively) in those with EGFR mutations. Backward elimination analyses revealed that age (OR = 0.93; 95% CI 0.89–0.98), GGO (OR = 0.14; 95% CI 0.03–0.67), and lymphadenopathy (OR = 4.15; 95% CI 1.49–11.60) were significantly associated with ALK rearrangement status. Conclusion: Our analyses revealed that clinical and CT characteristics of lung adenocarcinomas harboring ALK rearrangements were significantly different, compared with those with EGFR mutations. These differences may be related to the molecular pathology of these diseases.